PRIORITY CLAIM

This application claims priority to (i) U.S. Utility application Ser. No. 17/508,729 entitled “METHODS, COMPOSITIONS AND DEVICES FOR TREATING CANCER WITH ILLUDOFULVENES”, inventor: Michael Kelner filed Oct. 22, 2021 which claims priority to (ii) U.S. Utility application Ser. No. 16/708,005 entitled “METHODS, COMPOSITIONS AND DEVICES FOR TREATING CANCER WITH ILLUDOFULVENES”, inventor: Michael Kelner filed Dec. 9, 2019, which issued Nov. 2, 2021 as U.S. Pat. No. 11,160,807, which claims priority to (iii) U.S. Provisional Application No. 62/777,774 entitled “METHODS, COMPOSITIONS AND DEVICES FOR TREATING CANCER WITH ILLUDOFULVENES”, inventor: Michael Kelner filed Dec. 11, 2018, (iv) U.S. Provisional Application No. 62/796,409 entitled “METHODS, COMPOSITIONS AND DEVICES FOR TREATING CANCER WITH ILLUDOFULVENES”, inventor: Michael Kelner filed Jan. 24, 2019, and (v) U.S. Provisional Application No. 62/796,409 entitled “METHODS, COMPOSITIONS AND DEVICES FOR TREATING CANCER WITH ILLUDOFULVENES”, inventor: Michael Kelner filed Nov. 25, 2019 which applications (i)-(v) are herein expressly incorporated by reference in their entireties and for all purposes.

REFERENCE TO A “SEQUENCE LISTING,” A TABLE, OR A COMPUTER PROGRAM LISTING APPENDIX SUBMITTED AS AN ASCII TEXT FILE

The Sequence Listing written in file MKEL-01048US5_ST26.TXT, created on Jun. 26, 2024, 1,301,742 bytes, machine format IBM-PC, MS-Windows operating system, is hereby incorporated by reference.

TECHNICAL FIELD

The present invention relates to compositions, methods and devices for treating cancer with illudofulvenes including methods to detect the expression levels of genes encoding biomarkers in cancer patients and to predict the responsiveness of cancer patients to illudofulvenes.

BACKGROUND ART

DNA microarrays have been used to measure gene expression in samples from patients and to facilitate diagnosis of disease. Gene expression can reveal the presence of cancer in a patient in addition to the type, stage, and origin of the cancer. Gene expression may even have a role in predicting the efficacy of cancer therapies. In recent decades, the National Cancer Institute (NCI) has tested cancer therapeutics for their effect in limiting the growth of 60 human cancer cell lines. The NCI has also measured gene expression in those 60 cancer cell lines using DNA microarrays. Various studies have explored the relationship between gene expression and therapeutic effect using the NCI datasets.

Illudofulvenes have been proposed as medicants for Affinity Drug Conjugates in U.S. Pat. No. 10,285,955 issued May 14, 2019 to M. J. Kelner entitled Affinity Medicant Conjugates, which is incorporated by reference herein in its entirety and for all purposes.

SUMMARY OF INVENTION

There exists a continuing need for delivery of chemotherapeutic agents for which tumors do not have a medicant resistant phenotype and which inhibit tumor growth, especially solid tumor growth, and which have an adequate therapeutic index to be effective for in vivo treatment. The illudofulvenes of the present invention have shown efficacy in humans and animals for a wide range of therapeutic applications including: cancer, adenocarcinoma, carcinoma, breast cancer, prostate cancer, ovarian cancer, endometrial cancer, and neuroendocrine tumors. For example, veterinary and agricultural applications can include treatment of cancer, adenocarcinoma, carcinoma, ovarian cancer, endometrial cancer, and neuroendocrine tumors in farmyard and/or companion animals.

The invention features methods for detecting expression of a biomarker (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8, such as Excision Repair Cross-Complementation (ERCC) e.g., ERCC2 (SEQ ID NO:21)) in a patient having cancer, such as a human patient having cancer that is resistant to one or more cancer therapies other than an Illudofulvene Analog (e.g., a patient with prostate cancer, ovarian cancer, or hepatocellular carcinoma (HCC) that is resistant to one or more cancer therapies including radiation therapy and/or chemotherapy based on Other Drugs as set forth herein), and for determining responsiveness of a cancer patient (e.g., a patient with prostate cancer, ovarian cancer, or HCC) to treatment with an Illudofulvene Analog. The invention also features methods of treating cancer in a patient in need thereof (e.g., a patient with prostate cancer, ovarian cancer, or HCC or a treatment resistant form thereof) that include administering an Illudofulvene Analog to the patient, in which the patient is or has been determined to be responsive to an Illudofulvene Analog according to the diagnostic methods described herein. Without wishing to be bound by any theory, it is believed that sensitivity to Illudofulvene Analog treatment in patients can be caused by a deficiency in a component of the transcription-coupled repair (TCR) pathway or the post-replication repair pathway (e.g. ERCC or RAD18 E3 Ubiquitin Protein Ligase (RAD18)), or alternatively sensitivity to Illudofulvene Analogs can be caused by an upregulation or increase in a gene product (i.e., protein, e.g., MYC Proto-Oncogene (MYC), and/or Prostaglandin Reductase (PTGR), e.g., PTGR1, and/or PTGR2). A Southern blot, a northern blot, mRNA/RNA microarray, protein array, gene sequencing, immunohistochemical staining requiring additional tissue sampling and a readout by a competent pathologist can be utilized to identify the patient population.

The methods of this invention include predicting the sensitivity of illudofulvenes, preferably in the form of a pharmaceutical composition, to an animal in need thereof. In a further embodiment, pharmaceutical compositions are disclosed containing illudofulvenes of the present invention in combination with a pharmaceutically acceptable carrier.

In an embodiment of the present invention, an illudofulvenes can target cancer cells with reduced toxicity to normal cells. In an embodiment of the present invention, these compounds are useful in treatment of tumors in which a marker is expressed. In various embodiments of the present invention, pharmaceutical compositions comprising the illudofulvene compounds are used in the treatment of tumors in which the marker is expressed. In various embodiments of the present invention, methods of using the pharmaceutical compositions comprise these compounds to treat tumors in which the marker is expressed. In various embodiments of the present invention, pharmaceutical compositions comprising these compounds are used in the treatment of one or more of the following diseases: cancer, adenocarcinoma, carcinoma, breast cancer, prostate cancer, ovarian cancer, endometrial cancer.

BRIEF DESCRIPTION OF THE DRAWINGS

This invention is described with respect to specific embodiments thereof. Additional features can be appreciated from the Figures in which:

FIG. 1 shows the tumor regression produced by administration of an Illudofulvene Analog 002, according to an embodiment of the invention;

FIG. 2 shows that expression of MYC causes cancer cells to become sensitive to the illudofulvene (Analog 002), according to an embodiment of the invention;

FIG. 3 shows a comparison of Cell Count (%) as a function of the Illudofulvene Analog 002 concentration for two cell types (NIH3T3 MYC+ and NIH3T3 MYC-null), according to an embodiment of the invention;

FIG. 4A shows that MYC expression causes cells to become sensitive to Illudofulvene Analog 002, according to an embodiment of the invention;

FIG. 4B shows that MYC expression is not sensitive to Mithramycin;

FIG. 4C shows that MYC expression is not sensitive to Mitomycin C;

FIG. 4D shows that MYC expression is not sensitive to Topotecan; and

FIG. 5 shows a scheme relating the responsiveness of the cancer patient to treatment with Illudofulvenes based on the relative expression of the biomarkers.

DETAILED DESCRIPTION OF THE INVENTION

The term ‘upregulated’ means that the messenger RNA and/or the gene product (protein) is increased above basal level of a housekeeping gene by at least approximately one point nine (1.9) standard deviations which corresponds with ninety (90) percent confidence assuming a normal distribution. The phrase ‘increased expression’ means that the messenger RNA and/or the gene product (protein) is increased above a basal level of a housekeeping gene by at least approximately one point nine (1.9) standard deviations which corresponds with ninety (90) percent confidence assuming a normal distribution. The term ‘downregulated’ means that the messenger RNA and/or the gene product (protein) is decreased below basal levels of a housekeeping gene by at least approximately one point nine (1.9) standard deviations which corresponds with ninety (90) percent confidence assuming a normal distribution. The phrase ‘decreased expression’ means that the messenger RNA and/or the gene product (protein) is decreased below a basal level of a housekeeping gene by at least approximately one point nine (1.9) standard deviations which corresponds with ninety (90) percent confidence assuming a normal distribution. In relation to upregulation and/or downregulation, approximately means plus or minus ten (10) percent.

The phrase ‘not upregulated’ means that the messenger RNA and/or the gene product (protein) are within the normal range of the basal level of a housekeeping gene. A basal expression level is the expression level of an mRNA or a protein under normal circumstances. The phrase ‘determining if a cancerous tumor is sensitive to an illudofulvene analog’ means that the indicated gene is either upregulated or downregulated as specified. The phrase ‘using a selected technique’ means a technique selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

The phrase ‘determining the regulation of expression of a PTGR gene’ means probing the level of expression of the PTGR gene in the (tissue) sample using a selected technique. The phrase ‘determining the regulation of expression of a MYC gene’ means probing the level of expression of the MYC gene in the (tissue) sample using a selected technique. The phrase ‘determining the regulation of expression of a TCR gene’ means probing the level of expression of the TCR gene in the (tissue) sample using a selected technique. The phrase ‘determining the regulation of expression of a Housekeeping gene’ means probing the level of expression of the Housekeeping gene in the (tissue) sample using a selected technique. The phrase ‘determining a relative ratio of expression of a gene is high’ means calculating that the determined regulation of the gene is 1.9 standard deviations above the determined regulation of a housekeeping gene. The phrase ‘determining a relative ratio of expression of a gene is deficient’ means calculating that the determined regulation of the gene is 1.9 standard deviations below the determined regulation of a housekeeping gene.

The phrase ‘Other Drugs’ means docetaxel, cabazitaxel, mitoxantrone, estramustine, prednisone, carboplatin, bevacizumab, paclitaxel, gemcitabine, doxorubicin, topotecan, etoposide, tamoxifen, letrozole, sorafenib, fluorouracil, capecitabine, oxaliplatin, interferon-alpha, 5-fluorouracil (5-FU), a histone deacetylase (HDAC) inhibitor, ipilimumab, bortezomib, carfilzomib, thalidomide, lenalidomide, pomalidomide, dexamethasone, cyclophosphamide, vincristine, melphalan, tegafur, irinotecan, cetuximab, leucovorin, SN-38, everolimus, temsirolimus, bleomycin, lomustine, depsipeptide, erlotinib, cisplatin, busulfan, epirubicin, arsenic trioxide, bendamustine, fulvestrant, teniposide, adriamycin, decitabine, estramustine, azaguanine, aclarubicin, mitomycin, paclitaxel, taxotere, APO010, ara-c, methylprednisolone, methotrexate, methyl-gag, belinostat, idarubicin, IL4-PR38, valproic acid, all-trans retinoic acid (ATRA), cytoxan, suberoylanilide hydroxamic acid, leukeran, fludarabine, vinblastine, dacarbazine, hydroxyurea, tegafur, daunorubicin, mechlorethamine, streptozocin, carmustine, mercaptopurine, dactinomycin, tretinoin, ifosfamide, floxuridine, thioguanine, PSC 833, herceptin, celecoxib, iressa, anastrozole, and rituximab.

‘Acylfulvene’ means an Illudofulvene subgroup with the following structural molecular formula:

[Image Omitted]

where R₁, R₂, R₃, R₈, R₉, R₁₀, R₁₁, are as set forth in Formula P1, as given herein.

‘Formula P1’ means R₁represents —H, —CR₉R₈OH, —CHR₉CHR₈OH, —CHR₁₀CHR₉CHR₈OH, —CH₂CHR₁₀CHR₉CHR₈OH, —CR₉R₈OH, —CHR₉CHR₈OH, —CHR₁₀CHR₉CHR₈OH, —CH₂CHR₁₀CHR₉CHR₈OH, —C(═O)H, —CH₂(C═O)H, —CH₂CH₂(C═O)H, —CH₂CH₂CH₂(C═O)H, —CH₂CH₂CH₂CH₂(C═O)H, —CR₉R₈(C═O)H, —CHR₉CHR₈(C═O)H, —CH₂CHR₉CHR₈(C═O)H, —CR₉R₈(C═O)R₁₀, —CHR₉CHR₈(C═O)R₁₀, —CH₂CHR₉CHR₈(C═O)R₁₀, —CH₂CH₂CHR₉CHR₈(C═O)R₁₀, —CO₂H, —CHR₉CO₂H, —CHR₈CHR₉CO₂H, —CHR₁₀CHR₈CHR₉CO₂H, —CO₂R₁₀, —CHR₉CO₂R₁₀, —CHR₈CHR₉CO₂R₁₀, —CH₂CHR₈CHR₉CO₂R₁₀, —CHR₉CH₂CH₂CHR₈CO₂H, —CHR₉CH₂CH₂CHR₈CO₂R₁₀, —CR₈═CH₂, —CHR₈CH═CH₂, —CH₂CHR₈CH═CH₂, —CH₂CH₂CHR₈CH═CH₂, —CR₈═CHR₉, —CHR₈CR═CH₂, —CH₂CHR₈CR₉═CH₂, —CH₂CH₂CHR₈CR₉CH₂, —CR₈═CR₉R₁₀, —CHR₈CH═CR₉R₁₀, —CH₂CHR₈CH═CHR₉R₁₀, —CH₂CH₂CHR₈CH═CHR₉R₁₀, —Cl, —Br, —I, —F, —NO₂, —NR₈R₉, —CH₂Cl, —CH₂Br, —CH₂I, —CH₂F, —CR₈Cl₂, —CR₈Br₂, —CR₈I₂, —CR₈F₂, —CCl₃, —CBr₃, —CI₃, —CF₃, —CHR₈Cl, —CR₈R₉Cl, —CHR₈CHR₉Cl, —CHR₁₀CHR₈CHR₉Cl, —CH₂CHR₁₀CHR₈CHR₉Cl, —CHR₈Br, —CR₈R₉Br, —CHR₈CHR₉Br, —CHR₁₀CHR₈CHR₉Br, —CH₂CHR₁₀CHR₈CHR₉Br, —CHR₈I, —CR₈R₉I, —CHR₈CHR₉Br, —CHR₁₀CHR₈CHR₉I, —CH₂CHR₁₀CHR₈CHR₉I, —CR₈R₉NH₂, —CHR₈CHR₉NH₂, —CHR₁₀CHR₈CHR₉NH₂, —CH₂CH R₁₀CHR₈CHR₉NH₂—CR₉R₁₀NHR₈, —CHR₉CHR₁₀NHR₈, —CH₂CHR₉CHR₁₀NHR₈, —CH₂CH₂CHR₉CHR₁₀NHR₈, —CHR₉NHR₈R₁₀, —CHR₉CH₂NHR₈R₁₀, —CH₂CH₂CHR₉NR₁₀R₈, —CH₂CH₂CH₂CH₂NHR₈, —CR₉R₈OR₁₀, —CH₂CR₈R₉OR₁₀, —CH₂CH₂CR₈R₉OR₁₀, —CH₂CH₂CH₂R₈R₉OR₁₀, —CH₂CH₂CH₂CH₂R₈R₉OR₁₀, —CHR₈OC(═O)CHR₉R₁₀, —CHR8OC(═O)CHR₉R₁₀, —CH₂CH₂OC(═O)CHR₉R₁₀, —CH₂CH₂CHR₈OC(═O)CHR₉R₁₀, —CH₂CH₂CH₂CH₂OC(═O)CHR₉R₁₀, —CHR₈(═O)OCHR₉R₁₀, —CHR₈(═O)OCHR₉R₁₀, —CH₂CHR₈(═O)OCHR₉R₁₀, —CH₂CH₂CHR₈(═O)OCHR₉R₁₀, —CH₂CH₂CH₂CHR₈(═O)OCHR₉R₁₀, —R₁₂C═CR₉CH₂OH, —R₁₂C═CR₉C(═O)H, —R₁₂C═CR₉CH₂OR₁₀, —R₁₂C═CR₉C(═O)R₁₀, —CH₃, —CH₂CH₂, —CHR₈CH₂, —CHR₈CH₂CH₂, —CHR₈CHR₉CH₃, —OCH₃, —OCR₈R₉R₁₀, —OCH₂CR₈R₉R₁₀, —OCR₉R₈CHR₁₀, —OCHR₈CH₂CH₃, —OCHR₈CHR₉CH₃, —NR₈CH₃, —NR₈CH₂CH₃, —NR₉CHR₈CH₃, —NR₉CHR₈CH₂CH₃, —NR₁₀CHR₈CHR₉CH₃, —OCH₂OR₈, —OCHR₈OR₉, —OCHR₈CH₂OR₉, —OCHR₈CHR₉OR₁₀, —OC(═O)OR₈, —OCH₂C(═O)OR₈, —OCHR₉C(═O)OR₈, —CR₈(═N)H, —CH₂CR₈(═N)H, —CH₂CR₈(═N)H, —CH₂CH₂CR₈(═N)H, —CH₂CH₂CH₂CR₈(═N)H, —CH₂CH₂CH₂CH₂CR₈(═N)H, —CR₈(═N)OH, —CH₂CR₈(═N)OH, —CH₂CR₈(═N)OH, —CH₂CH₂CR₈(═N)OH, —CH₂CH₂CH₂CR₈(═N)OH, —CH₂CH₂CH₂CH₂CR₈(═N)OH, —CR₈(═N)R₉, —CH₂CR₈(═N)R₉, —CH₂CR₈(═N)R₉, —CH₂CH₂CR₈(═N)R₉, —CH₂CH₂CH₂CR₈(═N)R₉, —CH₂CH₂CH₂CH₂CR₈(═N)R₉, —CR₈(═N)OR₉, —CH₂CR₈(═N)OR₉, —CH₂CR₈(═N)OR₉, —CH₂CH₂CR₈(═N)OR₉, —CH₂CH₂CH₂CR₈(═N)OR₉, —CH₂CH₂CH₂CH₂CR₈(═N)OR₉, —CR₈(═N)NR₉, —CH₂CR₈(═N)NR₉, —CH₂CR₈(═N)NR₉, —CH₂CH₂CR₈(═N)NR₉, —CH₂CH₂CH₂—CR₈(═N)NR₉, —CH₂CH₂CH₂CH₂CR₈(═N)NR₉, —CR₈(═N)NR₉S(═O)₂R₁₀, —CH₂C(R₈)(═N)NR₉S(═O)₂R₁₀, —CH₂CH₂C(R₈)(═N)NR₉S(═O)₂R₁₀, —CH₂CH₂CH₂C(R₈)(═N)NR₉S(═O)₂R₁₀, CH₂CH₂CH₂CH₂C(R₈)(═N)NR₉S(═O)₂R₁₀, —R₁₂N(R₈)C(═O)NR₉R₁₀, —R₁₂N(R₈)C(═S)NR₉R₁₀, —R₁₂N(OR₈)C(═O)NR₉R₁₀, —R₁₂N(OR₈)C(═S)NR₉R₁₀, —R₁₂OS(O₂)NH₂, —R₁₂NHS(O₂)NH₂, —R₁₂OS(O₂)NR₈R₉, —R₁₂NHS(O₂)NR₈R₉, —CH₂N(R₈)S(O₂)NR₉R₁₀, —CH₂CH₂N(R₈)S(O₂)NR₉R₁₀, —CH₂CH₂CH₂N(R₈)S(O₂)NR₉R₁₀, —CH₂N(R₈)S(O₂)CR₉R₁₀R₁₁, —CH₂CH₂N(R₈)S(O₂) CR₉R₁₀R₁₁, —CH₂CH₂CH₂N(R₈)S(O₂) CR₉R₁₀R₁₁, —N(R₈)C(═O)R₉, —CH₂N(R₈)C(═O)R₉, —CH₂CH₂N(R₈)C(═O)R₉, —CH₂CH₂CH₂N(R₈)C(═O)R₉, —CH₂N(R₈)(C═O)NR₉R₁₀, —CH₂CH₂N(R₈)(C═O)NR₉R₁₀, —CH₂CH₂N(R₈)(C═O)NR₉R₁₀, —CH₂N(R₈)(C═O)CR₉R₁₀R₁₁, —CH₂CH₂N(R₈)(C═O)CR₉R₁₀R₁₁, —CH₂CH₂N(R₈)(C═O) CR₉R₁₀R₁₁, —R₁₂N(OH)C(═O)NHOH, —R₁₂N(OH)C(═S)NHOH, —R₁₂N(OR₈)C(═O)NHOR₉, —R₁₂N(OR₈)C(═S)NHOR₉, —R₁₂OS(O₂)NHOH, —R₁₂NHS(O₂)NHOH, —R₁₂OS(O₂)NHOR₉, —R₁₂OS(O₂)N(R₈)OR₉, —R₁₂NR₈S(O₂)NHOR₉, —CR₉(═N)OR₈, —NH(OR₈), —C(C═O)NHR₈, —C(C═O)NR₉R₈, —NR₁₀(OR₈)C(═O)R₉, —N(OR₈)C(═O)NR₉, —NR₈(R₉)S—, —N(R₈)S(═O)R₉, —NR(R₈)S(═O)₂R₉, —OC(═O)NR₈, —N(OR₈)C(═O)OR₉, —N(R₈)C(═S)R₉, —O(S(═O)₂R₈, —R₁₂O(S(═O)₂R₈, —O(S(═O)₂NR₈, —R₁₂O(S(═O)₂NR₈, —S(═O)R₈, —R₁₂S(═O)R₈, —S(═O)₂R₈, —R₁₂S(═O)₂R₈, —NR₁₀(R₉)S(═O)₂NHR₈, —NR₉(C═O)R₈, —NR₉(C═O)OR₈, —NR₉O(C═O)OR₈, —NR₉(C═O)NR₈R₁₀, —R₁₂N(R₉)S(═O)₂NHR₈, —R₁₂N(R₉)(C═O)R₈, —R₁₂N(R₉)(C═O)OR₈, —R₁₂N(R₉)(C═O)NR₈, —N(═NR₁₀)R₈, —R₁₂—N(═NR₁₀)R₈, —C(R₁₀)(═N—N═)CR₈R₉; —N₃, —CH₂N₃, —CH₂CH₂N₃, —CH₂CH₂CH₂N₃, —CH₂CH₂CH₂CH₂N₃, —CHR₈N₃, —CR₉R₈N₃, —CHR₉CHR₈N₃, —CH₂CHR₉CHR₈N₃, —CH₂CH₂CH₂CHR₈N₃, —C(R₈)═N—R₉, —CH₂C(R₈)═N—R₉, —CH₂CH₂C(R₈)═N—R₉, —CH₂CH₂CH₂C(R₈)═N—R₉, —N₃, —CH₂CH₂N₃, —CH₂CH₂CH₂N₃, —CH₂CH₂CH₂CH₂N₃; and R2 and R3 each independently represent —H, —OH, —CH₃, —OCH₃, —C(═O)CH₃, —OC(═O)CH₃, —C(═O)OCH₃, —C(═O)CH₂CH₃, —OC(═O)CH₂CH₃, —C(═O)OCH₂CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, —CH₂CH₂CH₃, —CH₂CH(CH₃)₂, —CHC(CH₃)₃, —CH₂OH, —NH₂, —CH₂NH₂, —N₃, and (C1-C4)alkyl; where R₈, R₉, R₁₀, R₁₁each independently represent —H, —OH, —CH₃, —CH₂CH₃, —CH₂═CH₂, —CH₂CH₂═CH₂, —CH₂CH₂CH₂═CH₂, —C(H)═O, —CH₂C(H)═O, —CH₂CH₂C(H)═O, —CH(CH₃)CH₃, —CH₂CH₂CH₃, —CH₂CH(CH₃)CH₃, —CHC(CH₃)₂CH₃, —C(═O)CH₃, —C(═O)CH₂CH₃, —C(═O)NH₂, —CH₂OH, —CH₂CH₂OH, —C(H)(OH)C(H₂)(OH), —OCH₃, —OC(CH₃)₃, —OCH₂CH₃, —OCH(CH₃)₂, —CO₂H, —C(═O)OCH₃, —C(═O)OCH₂CH₃, —OC(═O)CH₃, —OC(═O)CH₂CH₃, —OC(═O)OCH₃. —OC(═O)OCH₂CH₃, —C(CH₃)₃, —CH₂CH₂(CH₃)₂, —CHC(CH₃)₃, —CH₂CH₂OH, CH₂CH₂CH₂OH, —Cl, —Br, —I, —F, —CH₂Cl, —CH₂Br, —CH₂I, —CH₂F, —CHCl₂, —CHBr₂, —CHI₂, —CHF₂, —CCl₃, —CBr₃, —CI₃, —CF₃, —NH₂, —CH₂NH₂, —NH(OH), —CH₂N(OH), —NH(OCH₃), —N(OCH₂CH₃), —CH₂NH(OCH₃), —CH₂N(OCH₂CH₃), —N₃, —CH₂N₃, —CH₂CH₂N₃, —CH═NH, —CH═NOH, —CH═NOCH₃, —SH, —SCH₃, —SCH₂CH₃, —NO₂, —CN, cyclopropane ring, saturated or unsaturated cyclobutane ring, saturated or unsaturated cyclopentane ring, saturated or unsaturated cyclohexane ring, benzene ring, phenolic ring, xylene ring, an amino acid(s), (C₁-C₄)alkyl, and R₁₂represents —CH₂—, —CH₂CH₂—, —CH(CH₃)CH₂—, —C(CH₃)CH₂—, —CH═CH—, —O—, —S—, —O(C═O)—, —(C═O)O—, —NH—, —N(R₈)—, —N(OH)—, —CH₂—O—, —O—CH₂—, —CH₂CH₂—O—, —O—CH₂CH₂—, —S(═O)—, —(S═O)₂—, —NH(S═O)₂—, —N(OH)S(═O)₂—, —S(═O)₂NH—, —S(═O)₂N(OH)—, —NHS(═O)₂—, —N(OH)S(═O)₂—, —CH₂NH—, —CH₂N(R₈)—, —CH₂N(OH)—, —NHCH₂—, —N(R₈)CH₂—, —N(OH)CH₂—, —OC(C═O)O—, —OC(═O)NR₈—, —NR₈C(═O)O—.

An ‘Illudin’, which includes illudin M (where R₁, and R₃are CH₃, and R₂is OH), illudin S (where R₁is CH₂OH, R₂is OH, and R₃is CH₃) and syn-illudins (which include an illudin ring derivative (where R₂is the site of derivatization) or an illudin alkyl derivative (where R₁is the site of derivatization) means an Illudofulvene subgroup with the following structural molecular formula:

[Image Omitted]

where R₁, R₂, R₃, are as set forth in Formula P2, as given herein.

‘Formula P2’ means where R₁, R₂, and R₃each independently represent —H, —OH, —CH₃, —OCH₃, —C(═O)CH₃, —OC(═O)CH₃, —C(═O)OCH₃, —C(═O)CH₂CH₃, —OC(═O)CH₂CH₃, —C(═O)OCH₂CH₃, —CH₂CH₃, —CH(CH₃)₂, —C(CH₃)₃, —CH₂CH₂CH₃, —CH₂CH(CH₃)₂, —CHC(CH₃)₃, —CH₂OH, —NH₂, —NHOH, —CH₂NH₂, —CH₂NHOH, —N₃, and (C1-C4)alkyl.

Syn-Illudins are semi-synthetic derivatives of Illudins. Acylfulvenes are also semi-synthetic derivatives of Illudins. Syn-Illudins and Acylfulvenes have each been chemically modified at select sites to allow their use as medicants. The modifications in the Syn-Illudins do not alter any of the cyclic rings (cyclopropane, cyclopentane, cyclohexane) of the basic Illudin chemical structure. The modifications of Acylfulvenes differ from Syn-Illudins in that an additional double bond (an unsaturated bond) has been created in the 5 membered (cyclopentane) ring. Table 14 is a listing of IUPAC names of the Illudofulvene Analog according to various embodiments of the present invention.

During cancer treatment, critical time is often lost due to a trial and error approach to finding an effective therapy. In addition, cancer cells often develop resistance to a previously effective therapy. In such situations, patient outcome can be greatly improved by early detection of such resistance. Thus, there exists a need in the art for methods and devices that can predict the responsiveness of cancer patients to a medical treatment.

Exemplary types of cancer that can be diagnosed or treated with the methods include, e.g., prostate cancer, ovarian cancer (e.g., ovarian adenocarcinoma or embryonal carcinoma), liver cancer (e.g., HCC or hepatoma), myeloma (e.g., multiple myeloma), colorectal cancer (e.g., colon cancer and rectal cancer), leukemia (e.g., acute myeloid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, and chronic leukemia), myelodysplastic syndrome, lymphoma (e.g., diffuse large B-cell lymphoma, cutaneous T-cell lymphoma, peripheral T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, Waldenstrom's macroglobulinemia, and lymphocytic lymphoma), cervical cancer, esophageal cancer, melanoma, glioma (e.g., oligodendroglioma), pancreatic cancer (e.g., adenosquamous carcinoma, signet ring cell carcinoma, hepatoid carcinoma, colloid carcinoma, islet cell carcinoma, and pancreatic neuroendocrine carcinoma), gastrointestinal stromal tumor, sarcoma (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, leiomyosarcoma, Ewing's sarcoma, and rhabdomyosarcoma), breast cancer (e.g., medullary carcinoma), ER-positive cancer, bladder cancer, head and neck cancer (e.g., squamous cell carcinoma of the head and neck), lung cancer (e.g., non-small cell lung carcinoma, large cell carcinoma, bronchogenic carcinoma, and papillary adenocarcinoma), metastatic cancer, oral cavity cancer, uterine cancer, testicular cancer (e.g., seminoma and embryonal carcinoma), skin cancer (e.g., squamous cell carcinoma, and basal cell carcinoma), thyroid cancer (e.g., papillary carcinoma and medullary carcinoma), brain cancer (e.g., astrocytoma and craniopharyngioma), stomach cancer, intra-epithelial cancer, bone cancer, biliary tract cancer, eye cancer, larynx cancer, kidney cancer (e.g., renal cell carcinoma and Wilms tumor), gastric cancer, blastoma (e.g., nephroblastoma, medulloblastoma, hemangioblastoma, neuroblastoma, and retinoblastoma), polycythemia vera, chordoma, synovioma, mesothelioma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, cystadenocarcinoma, bile duct carcinoma, choriocarcinoma, epithelial carcinoma, ependymoma, pinealoma, acoustic neuroma, schwannoma, meningioma, pituitary adenoma, nerve sheath tumor, cancer of the small intestine, cancer of the endocrine system, cancer of the penis, cancer of the urethra, cutaneous or intraocular melanoma, a gynecologic tumor, solid tumors of childhood, or neoplasms of the central nervous system. For example, the cancer may be a solid tumor or a hematological cancer.

A first aspect of the invention features a method for detecting expression of a biomarker (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8, such as ERCC2 (SEQ ID NO:21)) in a patient having cancer (e.g., prostate cancer, ovarian cancer, or HCC), such as a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene (e.g., a patient with prostate cancer, ovarian cancer, or HCC that is resistant to one or more cancer therapies other than an illudofulvene). The method includes (a) contacting a sample (e.g., a tumor sample) from the patient including one or more nucleic acid molecules with a device (e.g., a microarray) including: i) one or more single-stranded nucleic acid molecules capable of specifically hybridizing with the nucleotides of one or more biomarkers of sensitivity selected from the biomarkers of Tables 1-8 (e.g., ERCC2 (SEQ ID NO:21)); and (b) detecting a level of expression of one or more of the biomarker(s) of sensitivity by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). Expression of the biomarker(s) may be detected by determining the level of a messenger RNA (mRNA) transcribed from a nucleic acid molecule corresponding to a gene of the biomarker (e.g., a mRNA expressed from the ERCC2 (SEQ ID NO:21)) or a complementary DNA (cDNA) thereof.

A second aspect of the invention features a method of determining responsiveness of a patient having cancer (e.g., one of the cancers noted above, such as prostate cancer, ovarian cancer, or HCC) to an illudofulvene. In particular, the patient may have a cancer that is resistant to one or more cancer therapies other than an illudofulvene, such as prostate cancer, ovarian cancer, or HCC that is resistant to one or more cancer therapies other than an illudofulvene. The method includes a) contacting a sample (e.g., a tumor sample) from the patient including one or more nucleic acid molecules with a device (e.g., a microarray or a device for performing a qRT-PCR reaction) including: i) one or more single-stranded nucleic acid molecules capable of specifically hybridizing with the nucleotides of one or more biomarkers of sensitivity selected from the biomarkers of Tables 1-8 (e.g., ERCC2 (SEQ ID NO:21)); and b) measuring hybridization, or an amplification product resulting from hybridization, between the one or more nucleic acid molecules from the patient and the single-stranded nucleic acid molecules of the device to detect a level of expression of one or more of the biomarkers of sensitivity. The patient is determined to be responsive to an illudofulvene if: i) the level of expression of the biomarkers of sensitivity is substantially similar to the level of expression of the biomarkers of sensitivity (e.g., ERCC2 (SEQ ID NO:21)) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive to an illudofulvene.

The method of the second aspect can further include administering an illudofulvene to the patient if: i) the level of expression of the biomarkers of sensitivity (e.g., ERCC2 (SEQ ID NO:21)) is substantially similar to the level of expression of the biomarkers of sensitivity in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive to an illudofulvene. The method can further include administering one or more cancer therapies other than an illudofulvene to the patient if: i) the level of expression of the biomarkers of sensitivity (e.g., ERCC2 (SEQ ID NO:21)) is substantially dissimilar to the level of expression of the biomarkers of sensitivity in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive to an illudofulvene.

In particular, the one or more of the cancer therapies includes surgery, radiation, or a therapeutic agent, such as Other Drugs as set forth herein.

The invention also features a method of treating cancer in a patient in need thereof (e.g., one of the cancers noted above, such as prostate cancer, ovarian cancer, or HCC) that includes administering an illudofulvene to the patient, in which the patient has been determined to be responsive to an illudofulvene according to the method of the first or second aspect of the invention. In particular, the patient may have a cancer that is resistant to one or more cancer therapies other than an Illudofulvene Analog selected from Table 14 (e.g., a patient with prostate cancer, ovarian cancer, or HCC that is resistant to one or more cancer therapies other than an illudofulvene).

A third aspect of the invention features a method of treating a patient having cancer (e.g., one of the cancers noted above, such as prostate cancer, ovarian cancer, or HCC). In particular, the patient may have a cancer that is resistant to one or more cancer therapies which use other than an Illudofulvene Analog selected from Table 14 (e.g., a patient with prostate cancer, ovarian cancer, or HCC that is resistant to one or more cancer therapies other than an illudofulvene). The method includes a) contacting a sample (e.g., a tumor sample) from the patient including one or more nucleic acid molecules with a device (e.g., a microarray or a device for performing a qRT-PCR reaction) including: i) one or more single-stranded nucleic acid molecules capable of specifically hybridizing with the nucleotides of one or more biomarkers of sensitivity selected from the biomarkers of Tables 1-8 (e.g., ERCC2 (SEQ ID NO:21)); and b) measuring hybridization, or an amplification product resulting from hybridization, between the one or more nucleic acid molecules from the patient and the single-stranded nucleic acid molecules of the device to detect a level of expression of one or more of the biomarkers of sensitivity and c) administering an Illudofulvene Analog selected from Table 14 to the patient if: i) the level of expression of the biomarkers of sensitivity is substantially similar to the level of expression of the biomarkers of sensitivity in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive to an illudofulvene.

The method of the third aspect of the invention may further include administering one or more additional therapies (e.g., surgery, radiation, or a therapeutic agent) to the patient prior to, concurrently with, or after administration of an Illudofulvene Analog selected from Table 14. In particular, the therapeutic agent may be selected from the group consisting of Other Drugs as set forth herein. The therapeutic agent can be administered parenterally (e.g. intravenously, intramuscularly, transdermally, intradermally, intra-arterially, intracranially, subcutaneously, intraorbitally, intraventricularly, intraspinally, intraperitoneally, or intranasally), enterally, or topically.

In the second or third aspect of the invention, an illudofulvene may be administered parenterally (e.g. intravenously, intramuscularly, transdermally, intradermally, intra-arterially, intracranially, subcutaneously, intraorbitally, intraventricularly, intraspinally, intraperitoneally, or intranasally), enterally, or topically. Preferably, an illudofulvene is administered by intravenous injection. An illudofulvene analog selected from Table 14 may be administered to the patient two or more times, such as one or more times daily, weekly, every two weeks, every three weeks, or monthly (e.g., one or more times per week for two weeks or more). Additionally, a second dose of an illudofulvene analog selected from Table 14 may be administered to the patient two weeks, three weeks, four weeks, or five weeks after administration of a prior dose of an illudofulvene. In particular, an illudofulvene analog selected from Table 14 is administered in a 3 week treatment regimen in which the illudofulvene analog selected from Table 14 is administered on day 1 an day 8. The treatment regimen may be repeated two to twenty times or more, as needed.

In particular, an illudofulvene analog selected from Table 14 may be administered to the patient at a dose of about 0.05 mg/kg to 6 mg/kg, such as at a dose of about 0.1 mg/kg to 1 mg/kg. For example, an illudofulvene may be administered at a dose of about 0.15 mg/kg, 0.20 mg/kg, 0.25 mg/kg, 0.30 mg/kg, 0.35 mg/kg, 0.40 mg/kg, 0.45 mg/kg, 0.55 mg/kg, 0.60 mg/kg, 0.65 mg/kg, 0.70 mg/kg, 0.75 mg/kg, 0.80 mg/kg, 0.85 mg/kg, 0.90 mg/kg, or 0.95 mg/kg. An illudofulvene may be administered to the patient in a treatment regimen at least once per week for at least two weeks and/or on day 1 and day 8 of a 3 week treatment regimen, in which the treatment regimen occurs one or more times (e.g., the treatment regimen is repeated two to twenty times). In particular, an illudofulvene is administered to the patient at a dose of about 0.1 mg/kg to 1 mg/kg, such as a dose of about 0.2 mg/kg to about 0.6 mg/kg (e.g., a dose of about 0.45 mg/kg). This dosage of an illudofulvene can be administered in a 3 week treatment regimen, in which an illudofulvene is administered on day 1 and day 8.

In the second or third aspect of the invention, the contacting step (a) and the measuring step (b) may occur prior to, concurrent with, or after administration of an illudofulvene analog selected from Table 14 to the patient. Additionally, the contacting step (a) and the measuring step (b) may occur two or more times, e.g., during treatment with an illudofulvene. For example, the contacting step (a) and the measuring step (b) may occur two or more times to assess the continued sensitivity of the patient to the Illudofulvene Analog selected from Table 14.

In any of the above aspects of the invention, the device can include at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, or more single-stranded nucleic acid molecules capable of specifically hybridizing with the nucleotides of one or more biomarkers of sensitivity selected from the biomarkers of Tables 1-8 (e.g., ERCC2 (SEQ ID NO:21)). Any of these aspects can be used individually or in any combination with other aspects. In particular, one or more of the single-stranded nucleic acid molecules of the device have a length in the range of 10 to 100 nucleotides in length (e.g., a length in the range of 20 to 60 nucleotides).

In any of the above aspects of the invention, the method may include converting the level of expression of one or more of the biomarkers of sensitivity (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8, such as ERCC2 (SEQ ID NO:21)) into a mean score, in which the mean score indicates the responsiveness of the patient to an Illudofulvene Analog selected from Table 14. In particular, the mean score and/or the difference score above a cutoff value indicates that the patient is responsive to an illudofulvene, such as if the cutoff value is above a specific numeric value.

In any of the above aspects of the invention, the biomarker of sensitivity may be selected from one or more of the ‘Select Sensitive Genes’, where the Select Sensitive Genes means ‘ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS:11-12), GTF2H5 (SEQ ID NOS:13-14), USP7 (SEQ ID NOS:15-16), UVSSA (SEQ ID NOS:17-18), ERCC3 (SEQ ID NOS:19-20), ERCC2 (SEQ ID NOS:21-22), ERCC4 (SEQ ID NOS:23-24), ERCC5 (SEQ ID NOS:25-26), XPA (SEQ ID NOS:27-28), RAD18 (SEQ ID NOS:29-30), ERCC6L (SEQ ID NOS:31-32), ERCC6L2 (SEQ ID NOS:33-34), MYC (SEQ ID NOS:222-223), ERCC1 (SEQ ID NOS:236-237), PTGR1 (SEQ ID NOS:249-250), and/or PTGR2 (SEQ ID NOS:262-263)’.

For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2) and ERCC6 (SEQ ID NOS:3-4). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:34), and GTF2H1 (SEQ ID NOS:5-6). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), and GTF2H2 (SEQ ID NOS:7-8). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8) and GTF2H3 (SEQ ID NOS:9-10). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:34), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10) and GTF2H4 (SEQ ID NOS:11-12). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS: 11-12) and GTF2H5 (SEQ ID NOS:13-14). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:34), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS:11-12), GTF2H5 (SEQ ID NOS:13-14), and USP7 (SEQ ID NOS:15-16). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:34), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS:11-12), GTF2H5 (SEQ ID NOS:13-14), USP7 (SEQ ID NOS:15-16), and UVSSA (SEQ ID NOS:17-18). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS:11-12), GTF2H5 (SEQ ID NOS:13-14), USP7 (SEQ ID NOS:15-16), UVSSA (SEQ ID NOS:17-18) and ERCC3 (SEQ ID NOS:19-20). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS:11-12), GTF2H5 (SEQ ID NO:13 or SEQ ID NO:14 or SEQ ID NO:101-111), USP7 (SEQ ID NOS:15-16), UVSSA (SEQ ID NOS:17-18), ERCC3 (SEQ ID NOS:19-20), and ERCC2 (SEQ ID NOS:21-22). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS: 11-12), GTF2H5 (SEQ ID NOS:13-14), USP7 (SEQ ID NOS:15-16), UVSSA (SEQ ID NOS:17-18), ERCC3 (SEQ ID NOS:19-20), ERCC2 (SEQ ID NOS:21-22), and ERCC4 (SEQ ID NOS:23-24). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS: 11-12), GTF2H5 (SEQ ID NOS:13-14), USP7 (SEQ ID NOS:15-16), UVSSA (SEQ ID NOS:17-18), ERCC3 (SEQ ID NOS:19-20), ERCC2 (SEQ ID NOS:21-22), ERCC4 (SEQ ID NOS:23-24) and ERCC5 (SEQ ID NOS:25-26). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS: 11-12), GTF2H5 (SEQ ID NOS:13-14), USP7 (SEQ ID NOS:15-16), UVSSA (SEQ ID NOS:17-18), ERCC3 (SEQ ID NOS:19-20), ERCC2 (SEQ ID NOS:21-22), ERCC4 (SEQ ID NOS:23-24), ERCC5 (SEQ ID NOS:25-26) and XPA (SEQ ID NOS:27-28). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS: 11-12), GTF2H5 (SEQ ID NOS:13-14), USP7 (SEQ ID NOS:15-16), UVSSA (SEQ ID NOS:17-18), ERCC3 (SEQ ID NOS:19-20), ERCC2 (SEQ ID NOS:21-22), ERCC4 (SEQ ID NOS:23-24), ERCC5 (SEQ ID NOS:25-26), XPA (SEQ ID NOS:27-28) and RAD18 (SEQ ID NOS:29-30). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS:11-12), GTF2H5 (SEQ ID NOS:13-14), USP7 (SEQ ID NOS:15-16), UVSSA (SEQ ID NOS:17-18), ERCC3 (SEQ ID NOS:19-20), ERCC2 (SEQ ID NOS:21-22), ERCC4 (SEQ ID NOS:23-24), ERCC5 (SEQ ID NOS:25-26), XPA (SEQ ID NOS:27-28), RAD18 (SEQ ID NOS:29-30) and ERCC6L (SEQ ID NOS:31-32). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS:11-12), GTF2H5 (SEQ ID NOS:13-14), USP7 (SEQ ID NOS:15-16), UVSSA (SEQ ID NOS:17-18), ERCC3 (SEQ ID NOS:19-20), ERCC2 (SEQ ID NOS:21-22), ERCC4 (SEQ ID NOS:23-24), ERCC5 (SEQ ID NOS:25-26), XPA (SEQ ID NOS:27-28), RAD18 (SEQ ID NOS:29-30), ERCC6L (SEQ ID NOS:31-32) and ERCC6L2 (SEQ ID NOS:33-34). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS:11-12), GTF2H5 (SEQ ID NOS:13-14), USP7 (SEQ ID NOS:15-16), UVSSA (SEQ ID NOS:17-18), ERCC3 (SEQ ID NOS:19-20), ERCC2 (SEQ ID NOS:21-22), ERCC4 (SEQ ID NOS:23-24), ERCC5 (SEQ ID NOS:25-26), XPA (SEQ ID NOS:27-28), RAD18 (SEQ ID NOS:29-30), ERCC6L (SEQ ID NOS:31-32), ERCC6L2 (SEQ ID NOS:33-34) and/or MYC (SEQ ID NOS:222-223). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS: 11-12), GTF2H5 (SEQ ID NOS:13-14), USP7 (SEQ ID NOS:15-16), UVSSA (SEQ ID NOS:17-18), ERCC3 (SEQ ID NOS:19-20), ERCC2 (SEQ ID NOS:21-22), ERCC4 (SEQ ID NOS:23-24), ERCC5 (SEQ ID NOS:25-26), XPA (SEQ ID NOS:27-28), RAD18 (SEQ ID NOS:29-30), ERCC6L (SEQ ID NOS:31-32), ERCC6L2 (SEQ ID NOS:33-34), MYC (SEQ ID NOS:222-223), and/or ERCC1 (SEQ ID NOS:236-237). For example, the biomarkers of sensitivity may include ERCC8 (SEQ ID NOS:1-2), ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS:11-12), GTF2H5 (SEQ ID NOS:13-14), USP7 (SEQ ID NOS:15-16), UVSSA (SEQ ID NOS:17-18), ERCC3 (SEQ ID NOS:19-20), ERCC2 (SEQ ID NOS:21-22), ERCC4 (SEQ ID NOS:23-24), ERCC5 (SEQ ID NOS:25-26), XPA (SEQ ID NOS:27-28), RAD18 (SEQ ID NOS:29-30), ERCC6L (SEQ ID NOS:31-32), ERCC6L2 (SEQ ID NOS:33-34), MYC (SEQ ID NOS:222-223), ERCC1 (SEQ ID NOS:236-237) and/or PTGR1 (SEQ ID NOS:249-250). For example, the biomarkers of sensitivity may include the Select Sensitive Genes.

In any of the above aspects of the invention, the device can be a microarray, such as a deoxyribonucleic acid (DNA)-based platform. Alternatively, the device can be for performing a qRT-PCR reaction (e.g., the device is used with a system for detecting the amplification product, for example, by fluorescence or by another method). The methods may also utilize both a microarray and a qRT-PCR. Thus, the expression level of the biomarkers of sensitivity (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8, such as ERCC2 (SEQ ID NO:21)) can be measured using qRT-PCR (Table 4). In particular, the level of expression of one or more of the biomarkers of sensitivity is determined by detecting the level of mRNA transcribed from one or more genes encoding one or more of the biomarkers of Tables 1-8.

In any of the above aspects of the invention, the cancer is selected from a solid tumor cancer and a hematological cancer. For example, the cancer is prostate cancer, ovarian cancer, HCC, multiple myeloma, breast cancer, acute myelogenous leukemia (AML), acute lympho-blastic leukemia (ALL), chronic lymphocytic leukemia (CLL), myelodysplastic syndrome (MDS), chronic myelogenous leukemia-chronic phase (CMLCP), diffuse large B-cell lymphoma (DLBCL), cutaneous T-cell lymphoma (CTCL), peripheral T-cell lymphoma (PTCL), Hodgkin's lymphoma, cervical cancer, renal cell carcinoma (RCC), esophageal cancer, melanoma, glioma, pancreatic cancer, gastrointestinal stromal tumors (GIST), sarcoma, estrogen receptor-positive (ERpos) breast cancer, non-small cell lung carcinoma (NSCLC), colon cancer, bladder cancer, or squamous cell carcinoma of the head and neck (SCCHN). In particular, the cancer is prostate cancer, such as prostate cancer that is resistant to one or more cancer therapies other than an illudofulvene. The cancer may also be ovarian cancer, such as ovarian cancer that is resistant to one or more cancer therapies other than an illudofulvene. Alternatively, the cancer may be HCC, such as HCC that is resistant to one or more cancer therapies other than an illudofulvene.

In any of the above aspects of the invention, the patient may exhibit cancer relapse (e.g., relapse of prostate cancer, ovarian cancer, or HCC), such as relapse after treatment with a therapeutic agent other than an illudofulvene. In particular, the patient may exhibit cancer relapse prior to treatment with an illudofulvene. Alternatively, the patient may have not been administered a treatment for cancer.

Described Methods

The expression levels of the biomarkers shown in Tables 1-8 in a patient having cancer are useful for predicting the responsiveness of the patient to an illudofulvene. These patients may already be determined to be resistance to a therapy other than an illudofulvene, such as Other Drugs as set forth herein.

A device, such as a microarray, with one or more single-stranded oligonucleotide probes that have substantial identity (e.g., at least 85%, 90%, 95%, 99%, or 100% sequence identity) to a sequence that is complementary or identical to the nucleic acid sequence of one or more biomarkers shown in Tables 1-8 can be used according to the methods described herein to assess the responsiveness of a cancer patient to treatment with an illudofulvene. For example, the probes can be used to detect one or more (e.g., two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8, such as ERCC2 (SEQ ID NO:21) or ERCC3 (SEQ ID NO:19) in a sample (e.g., a tumor sample) from a patient having cancer (e.g., prostate cancer, ovarian cancer, or hepatocellular carcinoma (HCC)).

Accordingly, the invention features individual biomarkers (e.g. ERCC2 (SEQ ID NO:21) or ERCC3 (SEQ ID NO:19)) and sets of biomarkers shown in Tables 1-8 that can be used to determine the responsiveness of a cancer patient to an illudofulvene at various stages of disease progression (e.g., patients diagnosed with cancer or patients after cancer recurrence) and at different times during the treatment process (e.g., prior to administration of any cancer treatment, after administration of one or more cancer treatments other than an illudofulvene, prior to administration of an illudofulvene, or during administration of an illudofulvene). Additionally, the methods can be used to determine the an illudofulvene responsiveness of a patient with cancer that is resistant to one or more cancer therapies other than an illudofulvene, such as Other Drugs as set forth herein.

The invention provides methods of determination for assessing whether a patient with a cancer may be responsive to treatment with illudofulvenes by detecting the expression magnitude of either mRNA, or the corresponding protein produced from this mRNA, of one or more of the biomarkers shown in Table 6 (e.g. ERCC2 (SEQ ID NO:21) in a biological sample (e.g. a direct tumor biopsy or alternatively collection of free circulating tumor cells in peripheral blood) obtained from the patient using a device (e.g. a cDNA/mRNA microarray or protein array). Table 6 also contains components of the TCR post-replication repair pathway, PTGR1, PTGR2, and Myc. The expression level of one or more of these biomarkers can be compared to the expression level of the same biomarker(s) in a cell or tissue known to be sensitive to illudofulvenes based on a known defect in the TCR-NER pathway. The patient may be responsive to illudofulvene treatment if the expression level of one or more of the biomarkers (e.g. the Select Sensitive Genes) are similar to the expression levels of the corresponding biomarker in a cell or tissue known to be sensitive to an Illudofulvene Analog.

The invention features methods for identifying biomarkers (e.g., one or more of the biomarkers of Tables 1-8) for determining the responsiveness of a cancer patient to a cancer treatment, such as an Illudofulvene Analog selected from Table 14. Such methods can involve, for example, an algorithm based on growth inhibition (GI50), tumor growth inhibition (TGI), and lethal concentration (LD50) values derived from an In vitro Cancer Growth Inhibition Screen that was performed using a tumor obtained from a patient that was subjected to treatment with an Illudofulvene. In addition, or alternatively, the expression of genes associated with the TCR-NER pathway in this tumor can be measured (e.g., using a microarray (e.g., an Affymetrix HG-U133A or HG-U133_Plus_2 array)) and their expression compared to expression of a control or housekeeping gene (e.g. beta actin (ACTB), transferrin receptor (TFRC), glyceraldehyde-3-phosphate dehydrogenase (GAPD), or ubiquitin) to determine the responsiveness of a cancer patient to a cancer treatment, such as an Illudofulvene.

The invention also features methods of treating a patient having cancer that is resistant to one or more cancer therapies other than treatment including an illudofulvene, by detecting the expression levels of one or more of the biomarkers shown in Table 6 (e.g. ERCC2 (SEQ ID NO:21) in a sample (e.g. a direct tumor biopsy or alternatively collection of free circulating tumor cells in peripheral blood) obtained from the patient and then administering an illudofulvene based on the expression levels of these biomarkers. In summary, a patient having cancer may be administered an illudofulvene if the expression level of one or more of the biomarkers of sensitivity are similar to the expression levels of the biomarker of sensitivity in a cell or tissue known to be sensitive to illudofulvenes. Thus, the methods described can be used to identify cancer patients who can be predicted to respond to illudofulvenes, such as patients having, e.g., prostate cancer, ovarian cancer, hepatocellular carcinoma (HCC), cervical cancer, renal cell carcinoma (RCC), esophageal cancer, melanoma, glioma, pancreatic cancer, gastrointestinal stromal tumors (GIST), sarcoma, estrogen receptor-positive (ERpos) breast cancer, non-small cell lung carcinoma (NSCLC), colon cancer, bladder cancer, squamous cell carcinoma of the head and neck (SCCHN), acute myelogenous leukemia (AML), acute lympho-blastic leukemia (ALL), chronic lymphocytic leukemia (CLL), myelodysplastic syndrome (MDS), chronic myelogenous leukemia-chronic phase (CMLCP), diffuse large B-cell lymphoma (DLBCL), cutaneous T-cell lymphoma (CTCL), peripheral T-cell lymphoma (PTCL), and Hodgkin's lymphoma. Alternatively, a patient having cancer would not be administered an illudofulvene if the expression levels of sensitivity of all biomarkers were substantially dissimilar to the expression level of biomarkers of sensitivity in a cell or tissue known to be sensitive to illudofulvenes.

Methods are described herein for identifying biomarkers of drug responsiveness, detecting this biomarker gene expression in samples from cancer patients, determining the responsiveness of a cancer patient to treatment with an illudofulvene, and treating cancer patients with an illudofulvene.

Methods of Determining the Responsiveness of a Patient to an Illudofulvene.

The invention features diagnostic methods for the detection and screening of cancer patients (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) that may be responsive to an illudofulvene analog selected from Table 14 using one or more of the biomarkers shown in Tables 1-8 (e.g. ERCC2 (SEQ ID NO:21) or ERCC3 (SEQ ID NO:19)). The methods of the invention may be used for predicting a patient's responsiveness to an illudofulvene, and optionally, treating the cancer patient throughout the progression of cancer and/or in cases of recurrence (e.g., after a first line treatment, a second line treatment, and/or a third line treatment).

The invention provides individual biomarkers (e.g. ERCC2 (SEQ ID NO:21) and sets of biomarkers (e.g., two or more of the biomarkers listed in Tables 1-8), the expression levels of which, as detected in a biological sample (e.g., a tumor sample, such as a biopsy) obtained from a cancer patient (e.g., a human with cancer), are indicative of responsiveness to an Illudofulvene Analog. The biomarkers were identified using methods similar to those previously described in, e.g., W Yang, et al. Genomics of drug sensitivity in cancer (GDSC): a resource for therapeutic biomarker discovery in cancer cells. Nucleic Acids Research, 41(D1): D955, 2013, which is incorporated by reference herein in its entirety for all purposes.

In particular, an algorithm based on the obtained in vitro cell culture values (i.e. growth inhibition values (GI50), or the total growth inhibition (TGI), or the lethal concentration (LC50)) of cell lines (e.g., NCI60 cells) subjected to treatment with an illudofulvene, and gene expression is determined in these same cell lines (e.g., by microarray analysis, reverse transcriptase polymerase chain reaction (RT-PCR), quantitative real-time PCR (qPCR), or next generation sequencing). After normalization, genes with, e.g., a Pearson correlation coefficient greater than about 0.5 or below about −0.5 to one of the obtained in vitro cell culture values, can be classified as biomarkers of sensitivity or resistance, respectively. A probability or significance value can also be calculated for the correlation between the obtained in vitro cell culture values (i.e. growth inhibition values (GI50), or the total growth inhibition (TGI), or the lethal concentration (LC50)) and the normalized expression of a specific gene.

In particular, a probability value of <0.05 is a statistically significant cut-off value as known in the art for establishing whether the expression level of A GENE, e.g., the genes shown in Tables 1-8, correlate with the likelihood of cancer treatment sensitivity, such as sensitivity to an illudofulvene. Thus, a probability value of <0.05 can be used to estimate the statistical significance of the expression level of the genes of Tables 1-8 for predicting patient responsiveness to treatment with an illudofulvene according to the methods described herein.

Biomarkers of Sensitivity.

The expression levels of one or more biomarkers of Tables 1-8 can be used to determine cancer patient responsiveness to treatment with an illudofulvene. Once determined to be responsive, the patient can be treated with an illudofulvene. In particular, the biomarker ERCC8 (SEQ ID NO:1) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker ERCC8 (SEQ ID NO:1) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of ERCC8 (SEQ ID NO:1) in the patient sample may then be compared, e.g., to the expression level of ERCC8 (SEQ ID NO:1) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker ERCC8 (SEQ ID NO:1) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as ERCC6 (SEQ ID NOS:3-4), GTF2H1 (SEQ ID NOS:5-6), GTF2H2 (SEQ ID NOS:7-8), GTF2H3 (SEQ ID NOS:9-10), GTF2H4 (SEQ ID NOS:11-12), GTF2H5 (SEQ ID NOS:13-14), USP7 (SEQ ID NOS:15-16), UVSSA (SEQ ID NOS:17-18), ERCC3 (SEQ ID NOS:19-20), ERCC2 (SEQ ID NOS:21-22), ERCC4 (SEQ ID NOS:23-24), ERCC5 (SEQ ID NOS:25-26), XPA (SEQ ID NOS:27-28), RAD18 (SEQ ID NOS:29-30), ERCC6L (SEQ ID NOS:31-32), ERCC6L2 (SEQ ID NOS:33-34), MYC (SEQ ID NOS:222-223), ERCC1 (SEQ ID NOS:236-237), PTGR1 (SEQ ID NOS:249-50), and/or PTGR2 (SEQ ID NOS:262-263). The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The expression level of the biomarker ERCC6 (SEQ ID NO:3) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of ERCC6 (SEQ ID NO:3) in the patient sample may then be compared, e.g., to the expression level of ERCC6 (SEQ ID NO:3) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker ERCC6 (SEQ ID NO:3) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker GTF2H1 (SEQ ID NO:5) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker GTF2H1 (SEQ ID NO:5) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of GTF2H1 (SEQ ID NO:5) in the patient sample may then be compared, e.g., to the expression level of GTF2H1 (SEQ ID NO:5) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker GTF2H1 (SEQ ID NO:5) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker GTF2H2 (SEQ ID NO:7) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker GTF2H2 (SEQ ID NO:7) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of GTF2H2 (SEQ ID NO:7) in the patient sample may then be compared, e.g., to the expression level of GTF2H2 (SEQ ID NO:7) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker GTF2H2 (SEQ ID NO:7) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker GTF2H3 (SEQ ID NO:9) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker GTF2H3 (SEQ ID NO:9) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of GTF2H3 (SEQ ID NO:9) in the patient sample may then be compared, e.g., to the expression level of GTF2H3 (SEQ ID NO:9) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker GTF2H3 (SEQ ID NO:9) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker GTF2H4 (SEQ ID NO:11) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker GTF2H4 (SEQ ID NO:11) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of GTF2H4 (SEQ ID NO:11) in the patient sample may then be compared, e.g., to the expression level of GTF2H4 (SEQ ID NO:11) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker GTF2H4 (SEQ ID NO:11) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker GTF2H5 (SEQ ID NO:13) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker GTF2H5 (SEQ ID NO:13) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of GTF2H5 (SEQ ID NO:13) in the patient sample may then be compared, e.g., to the expression level of GTF2H5 (SEQ ID NO:13) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker GTF2H5 (SEQ ID NO:13) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker USP7 (SEQ ID NO:15) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker USP7 (SEQ ID NO:15) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of USP7 (SEQ ID NO:15) in the patient sample may then be compared, e.g., to the expression level of USP7 (SEQ ID NO:15) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker USP7 (SEQ ID NO:15) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker UVSSA (SEQ ID NO:17) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker UVSSA (SEQ ID NO:17) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of UVSSA (SEQ ID NO:17) in the patient sample may then be compared, e.g., to the expression level of UVSSA (SEQ ID NO:17) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker UVSSA (SEQ ID NO:17) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker ERCC3 (SEQ ID NO:19) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker ERCC3 (SEQ ID NO:19) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of ERCC3 (SEQ ID NO:19) in the patient sample may then be compared, e.g., to the expression level of ERCC3 (SEQ ID NO:19) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker ERCC3 (SEQ ID NO:19) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker ERCC2 (SEQ ID NO:21) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker ERCC2 (SEQ ID NO:21) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of ERCC2 (SEQ ID NO:21) in the patient sample may then be compared, e.g., to the expression level of ERCC2 (SEQ ID NO:21) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker ERCC2 (SEQ ID NO:21) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker ERCC4 (SEQ ID NO:23) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker ERCC4 (SEQ ID NO:23) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of ERCC4 (SEQ ID NO:23) in the patient sample may then be compared, e.g., to the expression level of ERCC4 (SEQ ID NO:23) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker ERCC4 (SEQ ID NO:23) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker ERCC5 (SEQ ID NO:25) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker ERCC5 (SEQ ID NO:25) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of ERCC5 (SEQ ID NO:25) in the patient sample may then be compared, e.g., to the expression level of ERCC5 (SEQ ID NO:25) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker ERCC5 (SEQ ID NO:25) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker XPA (SEQ ID NO:27) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker XPA (SEQ ID NO:27) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of XPA (SEQ ID NO:27) in the patient sample may then be compared, e.g., to the expression level of XPA (SEQ ID NO:27) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker XPA (SEQ ID NO:27) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker RAD18 (SEQ ID NO:29) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker RAD18 (SEQ ID NO:29) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of RAD18 (SEQ ID NO:29) in the patient sample may then be compared, e.g., to the expression level of RAD18 (SEQ ID NO:29) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker RAD18 (SEQ ID NO:29) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker ERCC6/L (SEQ ID NO:31) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker ERCC6/L (SEQ ID NO:31) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of ERCC6/L (SEQ ID NO:31) in the patient sample may then be compared, e.g., to the expression level of ERCC6/L (SEQ ID NO:31) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker ERCC6/L (SEQ ID NO:31) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker ERCC6/L2 (SEQ ID NO:33) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker ERCC6/L2 (SEQ ID NO:33) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of ERCC6/L2 (SEQ ID NO:33) in the patient sample may then be compared, e.g., to the expression level of ERCC6/L2 (SEQ ID NO:33) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker ERCC6/L2 (SEQ ID NO:33) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker MYC (SEQ ID NO:222) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker MYC (SEQ ID NO:222) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of MYC (SEQ ID NO:22) in the patient sample may then be compared, e.g., to the expression level of MYC (SEQ ID NO:222) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker MYC (SEQ ID NO:222) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker ERCC1 (SEQ ID NO:236) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker ERCC1 (SEQ ID NO:236) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of ERCC1 (SEQ ID NO:236) in the patient sample may then be compared, e.g., to the expression level of ERCC1 (SEQ ID NO:236) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker ERCC1 (SEQ ID NO:236) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker PTGR1 (SEQ ID NO:249) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker PTGR1 (SEQ ID NO:249) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of PTGR1 (SEQ ID NO:249) in the patient sample may then be compared, e.g., to the expression level of PTGR1 (SEQ ID NO:249) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker PTGR1 (SEQ ID NO:249) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The biomarker PTGR2 (SEQ ID NO:262) may be used to assess a cancer patient's (e.g., a patient having cancer that is resistant to one or more cancer therapies other than an illudofulvene) responsiveness to an illudofulvene. The expression level of the biomarker PTGR2 (SEQ ID NO:262) may be assessed using nucleic acid amplification methods (e.g., PCR) or a device (e.g., a microarray). As is described above, the expression level of PTGR2 (SEQ ID NO:262) in the patient sample may then be compared, e.g., to the expression level of PTGR2 (SEQ ID NO:262) in a cell (e.g., a cancer cell) or tissue (e.g., a tumor tissue) known to be sensitive or resistant to treatment with an illudofulvene and used to determine the cancer patient's responsiveness to an illudofulvene. The biomarker MYC (SEQ ID NO:222) may be used alone to predict cancer patient responsiveness to treatment with an illudofulvene or in combination with one or more additional biomarkers (e.g., one, two, three, four, five, ten, or all of the biomarkers shown in Tables 1-8), such as the Select Sensitive Genes. The expression level of the biomarker(s) may be determined using, e.g., a microarray, PCR, or other techniques described herein, for example, using a nucleic acid probe sequence based on the target sequences shown in Tables 1-8.

The following Examples 1-4, are intended to illustrate, rather than limit, the invention.

Example 1

Identification of Biomarkers of Sensitivity to an Illudofulvene using cell lines deficient in a specific DNA repair gene. Cell growth inhibition assays were performed using Illudofulvene against a bank of cell lines which are deficient in DNA repair based on lack of expression of a specific DNA repair gene (Table 21).

TABLE 21

Sensitivity of DNA repair Deficient Cells to Illudofulvene Analog

GI50

DNA-
value (nM)
Corresponding

repair Gene
Mean +/ SD
GENE

Cell line
Deficient
(N = 3)
SEQ ID NOS:

AA8
None (wild type)
1440 ± 70
None

UV20
ERCC1
144 ± 4
236-237

UV5
ERCC2
60 ± 7
21, 22, 145-155

UV24
ERCC3
127 ± 13
19, 20, 134-144

UV41
ERCC4
167 ± 10
23, 24, 156-166

UV135
ERCC5
174 ± 22
25, 26, 167-177

UV61
ERCC6
366 ± 18
3, 4, 46-56

EM9
XRCC1
1197 ± 44
None

Studies clearly identified that a cell which is deficient in Transcription-coupled repair (TCR) (e.g. ERCC2 or ERCC3) is sensitive to Illudofulvene as compared to its parental line (AA8) whereas cells deficient in other DNA repair components, such as homologous recombination (e.g. XRCC1). These results are consistent with the finding that Illudofulvene Analogs produce a damage that is repaired only by the TCR genes and that other DNA-repair pathways (e.g. homologous recombination, Base-excision, etc.) are not involved (Kelner, et al. Cancer Research, 55:4936-40, 1995; and Jaspers, et al. DNA Repair 1:1027-38, 2002, both of which are herein expressly incorporated by reference in their entireties and for all purposes). TCR genes means Excision Repair Cross-Complementation (ERCC) 1 (ERCC1), ERCC2, ERCC3, ERCC4, ERCC5, ERCC6, ERCC6L, ERCC6L2, DNA Damage Recognition and Repair Factor (XPA), Ubiquitin Specific Peptidase7 (USP7), UV Stimulated Scaffold Protein A (UVSSA), E3 ubiquitin-protein ligase RAD18 (RAD18), General Transcription Factor IIH (GTF2H) Subunit 1 (GTF2H1), GTF2H2, GTF2H2, GTF2H3, GTF2H4, GTF2H5 genes.

Example 2

Identification of Biomarkers of Sensitivity to An Illudofulvene Using Gene Expression data derived by DNA chip analysis of the 60 cancer cell lines of the NCI60 data set.

The expression of thousands of genes in each of the cell lines composing the NCI DTP 60 cell line has been analyzed using a variety of different DNA chips (or microarrays) and is publicly available at website dtp.cancer.gov/compare-web-public_compare/SearchAndDisplay/SearchAndDisplay.

Data has been generated using a variety of Affymetrix DNA chips including HG-U133A array (GENELOGIC_133 data set), HG-U133 Plus 2.0 array (GENELOGIC_133PLUS2 data set), the HG-95 (GENELOGIC_U95 data set), as well as others. Results from DNA microchips produced by other companies, e.g. Novartis, are also available. The data from these array experiments has been log it normalized to allow a direct comparison of a genes expression between different cell lines. For each array, the logit transformation was performed followed by a Z-transformation to mean zero and SD.

One can determine the growth inhibition (GI50), total growth inhibition (TGI), or lethal concentration (LC50) for a drug for each of the cell lines in the NCI DTP 60 cell line panel. One can use these 3 values to search for correlations against all gene expressions in all cell lines using the NCI's publicly available CELL MINER Software. Alternatively, one can down load for a specific gene its relative expression in each of the 60 cell lines using the NCI's publicly available COMPARE Software, and performs the expression for a specific gene.

A gene's expression in each cell line can then be correlated to the ability of an illudofulvene to hinder cell growth (GI50, TGI, or LC50 value) and a correlation coefficient determined to identify genes positively and negatively correlated to sensitivity to an Illudofulvene Analog. Positively correlated genes can be those in which as the expression of the gene increase, the tumor cells become sensitive to the drug. Negatively correlated genes can be those that as the expression of the gene decreases, the tumor cells become sensitive to the drug.

Analysis of the cell growth inhibition values for the Illudofulvene Analog 002, to specific gene expression values retrieved by COMPARE from data generated by the Affymetrix HG-U133 Plus 2.0 array again confirms that a deficiency in expression of components of TCR-pathway confers sensitivity to Illudofulvene Analog 002 (Table 16) including ERCC2, ERCC8, GTF2H1, USP7, and XPC. However, there was no correlation with components of other DNA repair pathways such as double strand repair (RAD50), NER (XPC), X-ray repair (XRCC1), etc.

Similar results are obtained with illudofulvene Analogs 142, 176, 184, (see Tables 17-20).

Example 3

Identification of Biomarkers whose expression causing sensitivity to Illudofulvene Analog 002 followed by confirmation using Gene Expression data derived by DNA chip analysis of the 60 cancer cell lines of the NCI60 data set (see, Example 2 above) and also confirmed using cell lines deficient in a specific GENE (see, Example 1 above).

To further explore the mechanism(s) of action of Illudofulvene Analog 002, we used phage display technology to identify peptides capable of binding to the drug and utilized this information to identify potential target proteins for Illudofulvene Analog 002 binding. The proto-oncogene c-myc (Gene Symbol MYC) was identified as a putative target, and its role in sensitizing tumor cells to an Illudofulvene Analog was confirmed using Gene Expression data derived by DNA chip analysis of the 60 cancer cell lines of the NCI60 data set (see, Example 2 above) and then confirmed in vitro using cell lines deficient in a specific GENE (see, Example 1 above).

Three different ER2537-derived phage libraries (New England Biolabs, Beverly, MA) were used for biopanning studies; 7-mer, 12-mer, and C7C. The latter consisted of a 7-mer library flanked by two cysteines to allow disulfide formation. The aldehyde derivative of Illudofulvene Analog 002 was prepared by reacting Illudofulvene Analog 001 with acrolein, and then linked to biotin using Biotin-LC-hydrazide (Pierce). A 96 well avidin coated plate was coated with 75 μg of the target molecule (Illudofulvene Analog 001-biotin conjugate) in 150 μl of PBS, and rocked gently overnight at 4° C. The desired phage library was added to the wells and the plate gently rocked for 60 minutes at room temperature. Wells were rinsed ten times with TBST (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.1% v/v tween-20). Bound phage was eluted by applying excess target molecule (Illudofulvene Analog 001) and increasing salt concentration. One μl of the eluate was used for titering and the remaining eluate was amplified using standard techniques.

Using phage display panning we isolated 157 nonredundant sequences capable of binding to Illudofulvene Analog 002. Two algorithms were used to predict a sequence and 3D structure of a protein capable of binding to Illudofulvene Analog 002; “peptide elongation without substitution” and “peptide elongation with substitution”. Each of the 157 sequences was initially aligned using a FASTA program to proteins in the PIR database followed by the desired peptide elongation process. A complete description of the process has been published (Tsigelny I, Sharikov Y, Kelner M J, and Ten Eyck L F., Protein structure prediction on the basis of combinatorial peptide libraries screening. In: Protein Structure Prediction: A Bioinformatics Approach. Ed: I Tsigelny. International University Line Publishing, 2001, which is herein expressly incorporated by reference in its entirety for all purposes). The algorithm of elongation without substitution identified c-myc as a potential target of Illudofulvene Analog 002 and docking studies revealed a potential binding site for Illudofulvene Analog 002 that matched the peptides recovered by phage panning.

To determine if the MYC gene had clinical significance in regards to the therapeutic activity of Illudofulvene Analog 002, we inputted the in vitro cell culture values (i.e. growth inhibition values (GI50), or the total growth inhibition (TGI), or the lethal concentration (LC50) obtained for Illudofulvene Analog 002 from cell lines contained in the NCI 60 cell line panel and used CELLMINER to screen for correlation against the normalized expression of genes in cell lines in the NCI 60 cell line panel.

Using CELLMINER, after normalization, genes with, e.g., a Pearson correlation coefficient greater than about 0.2 or below about −0.2 can be classified as biomarkers of sensitivity or resistance, respectively. In particular, a correlation coefficient of about 0.2 or greater is a statistically significant cut-off known in the art for establishing whether the expression level of A GENE, e.g., the genes shown in Tables 1-8, correlate with the likelihood of cancer treatment sensitivity, such as sensitivity to Illudofulvene Analog 002. Thus, a correlation coefficient of about 0.2 or greater or about −0.2 or lower can be used to estimate the statistical significance of the expression level of the genes of Tables 1-8 for predicting patient responsiveness to treatment with an Illudofulvene Analog according to the methods described herein.

Using CELLMINER the correlation of MYC expression to Illudofulvene Analog 002 activity was identified at 0.42, and a p value of 0.000941 (highly significant) indicating MYC expression in a patient's tumor can be predictive of response to treatment with Illudofulvene Analog 002. A similar significant correlation between MYC expression and GI50 values for CAP-0142 (P<0.001) and CAP-0241 (P<0.001). To confirm the role of c-myc in Illudofulvene Analog 002 cytotoxicity, studies were conducted using the c-myc negative Rat 1A line, and c-myc positive daughter line Myc/Rat1A. The expression of c-myc in this cell line is under the control of a modified receptor that responds to 4-hydroxytamoxifen (4HOT). FIG. 2 shows that the presence of 4HOT expresses c-myc and induces sensitivity to expression of MYC causing cancer cells to become sensitive to illudofulvene (Analog 002) in the Myc/ER daughter, but not the Rat 1A parental cell line.

FIG. 3 shows the influence of c-myc expression on Illudofulvene Analog 002 cytotoxicity in a comparison of Cell Count (%) as a function of Analog 002 concentration for a paired c-myc expressing (NIH3T3 MYC+) cell line and a c-myc nonexpressing (NIH3T3) or null cell line. Expression of cellular c-myc conferred a greater than 10 fold increase in sensitivity to illudofulvene Analog 002 to the daughter NIH3T3 MYC+ cell line as compared to the parental c-myc null or nonexpressing NIH3T3 cell line, based on the decrease in cell count. The ability of c-myc expression to alter cellular sensitivity to other agents was also studied. The expression of c-myc, however, did not alter cellular sensitivity towards a variety of other anticancer agents (including mithramycin, mitomycin C, topotecan (as shown in FIG. 4B-FIG. 4D) or cisplatin (data not shown) as compared to Illudofulvene Analog 002 (see FIG. 4A).

Example 4

Most relevant to clinical applications, Illudofulvene Analog 002 activity is independent of common resistance mechanisms such as the multi-medicant resistance phenotype, anti-apoptotic B-cell lymphoma 2 (Bcl-2) over expression, as well as tumor protein 53 (p53) and cyclin dependent kinase inhibitor 1 (p21/WAF1) mutations (see Tables 7 and 11).

Growth factors, including peptides and proteins are critical mediators of a wide range of cell-cell communication. They are important endocrine, paracrine and autocrine messengers. Growth factors function as neurotransmitters and neuromodulators, regulate chemotaxis, immune function, development, cell growth, and can influence tumor cells. The receptors that recognize growth factors are highly selective and define specific cell populations. As a result, growth factor receptors are a large and important class of medicant (including drug) targets. In addition to physiologic noncancerous cell populations, these receptors can also be expressed in various cancer cell populations.

FIG. 1 shows the ability of the illudofulvene Analog 002 to induce tumor regression in the MV522 cell line (a lung-derived adenocarcinoma cell line) compared with other alternative compounds (i.e., Fludarabine, Cytarabine and Doxorubicin). In various embodiments of the invention, the MV522 cell line represents a “target” cell line. That is an illudofulvene analog that exhibits toxicity against this solid tumor cell line shows a desirable result. The 8392B cell line represents a hematopoietic cell line. In various embodiments of the invention, the 8392B cell line is considered a “nontarget” cell line. The two hour toxicity data represents the concentration of a given analog for which a two hour exposure will inhibit 50% of the DNA synthesis activity in a given cell line. The 48 hour exposure data represents the concentration at which a given analog with a 48 hour exposure will inhibit the growth or viability in a given cell line as defined by the standard Trypan Blue Exclusion assay. As an example, Illudofulvene Analog 002 will inhibit the target MV522 cell line at 110 nM with only a 2 hour exposure but has no inhibitory effect on the nontarget 8392B cell line at 26,000 nM (26 μM), see Table 13. Illudofulvene Analog 002 with a prolonged exposure period (e.g. 48 hours) can eventually inhibit the 8392B nontarget cell line, see Table 13. In contrast, Illudofulvene Analog 201 will inhibit the target MV522 cell line with only a 2 hour exposure (IC50=360 nM) but has minimal effect on the 8392B cell nontarget line with even a 48 hour exposure (IC50=26.000 nM) indicating superior anticancer activity as a monotherapeutic agent, see Table 13. In contrast to these two analogs. Illudofulvene Analog 224 displayed minimal toxicity as well as no differential toxicity between the target and nontarget cell line indicating it can have minimal properties as a monotherapeutic anticancer agent, see Table 13. Table 10 indicates that Illudofulvene Analog 001 or Illudofulvene Analog 002 have a novel mechanism of action versus chemotherapeutic agents including Other Drugs as set forth herein.

Compositions and Methods of Administration.

In other embodiments, described is a pharmaceutical composition including an effective amount of an illudofulvene and a pharmaceutically acceptable carrier or vehicle. The compositions are suitable for veterinary or human administration.

The present pharmaceutical compositions can be in any form that allows for the composition to be administered to a patient. For example, the composition can be in the form of a solid or liquid. Typical routes of administration include, without limitation, parenteral, ocular and intra-tumor. Parenteral administration includes subcutaneous injections, intravenous, intramuscular or intrasternal injection or infusion techniques. In one aspect, the compositions are administered parenterally. In a specific embodiment, the compositions are administered intravenously.

Pharmaceutical compositions can be formulated so as to allow an illudofulvene to be bioavailable upon administration of the composition to a patient. Compositions can take the form of one or more dosage units, where for example, a tablet can be a single dosage unit, and a container of an illudofulvene in liquid form can hold a plurality of dosage units.

Materials used in preparing the pharmaceutical compositions can be non-toxic in the amounts used. It will be evident to those of ordinary skill in the art that the optimal dosage of the active ingredient(s) in the pharmaceutical composition will depend on a variety of factors. Relevant factors include, without limitation, the type of animal (e.g., human), the particular form of the illudofulvene, the manner of administration, and the composition employed.

The pharmaceutically acceptable carrier or vehicle can be solid or particulate, so that the compositions are, for example, in tablet or powder form. The carrier(s) can be liquid. In addition, the carrier(s) can be particulate.

The composition can be in the form of a liquid, e.g., a solution, emulsion or suspension. In a composition for administration by injection, one or more of a surfactant, preservative, wetting agent, dispersing agent, suspending agent, buffer, stabilizer and isotonic agent can also be included.

The liquid compositions, whether they are solutions, suspensions or other like form, can also include one or more of the following: sterile diluents such as water for injection, saline solution, preferably physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils such as synthetic mono or diglycerides which can serve as the solvent or suspending medium, polyethylene glycols, glycerin, cyclodextrin, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediamine tetraacetic acid; buffers such as acetates, citrates, phosphates or amino acids and agents for the adjustment of tonicity such as sodium chloride or dextrose. A parenteral composition can be enclosed in ampoule, a disposable syringe or a multiple-dose vial made of glass, plastic or other material. Physiological saline is an exemplary adjuvant. An injectable composition is preferably sterile.

The amount of the illudofulvene that is effective in the treatment of a particular disorder or condition will depend on the nature of the disorder or condition, and can be determined by standard clinical techniques. In addition, in vitro or in vivo assays can optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the compositions will also depend on the route of administration, and the seriousness of the disease or disorder, and can be decided according to the judgment of the practitioner and each patient's circumstances.

The compositions comprise an effective amount of an illudofulvene such that a suitable dosage will be obtained. Typically, this amount can be at least about 0.01% of an illudofulvene by weight of the composition. In an exemplary embodiment, pharmaceutical compositions are prepared so that a parenteral dosage unit (actual powder) dissolved and used to inject the patient was 98% drug by weight, although once diluted in an IV the dose contains from about 0.01% to about 2% by weight).

For intravenous administration, the composition can comprise from about 0.01 to about 100 mg of an illudofulvene per kg of the patient's body weight. In one aspect, the composition can include from about 1 to about 100 mg of an illudofulvene per kg of the patient's body weight. In another aspect, the amount administered will be in the range from about 0.1 to about 25 mg/kg of body weight of the illudofulvene.

Generally, the dosage of an illudofulvene administered to a patient is typically about 0.01 mg/kg to about 20 mg/kg of the patient's body weight. In one aspect, the dosage administered to a patient is between about 0.01 mg/kg to about 10 mg/kg of the patient's body weight. In another aspect, the dosage administered to a patient is between about 0.1 mg/kg and about 10 mg/kg of the patient's body weight. In yet another aspect, the dosage administered to a patient is between about 0.1 mg/kg and about 5 mg/kg of the patient's body weight. In yet another aspect the dosage administered is between about 0.1 mg/kg to about 3 mg/kg of the patient's body weight. In yet another aspect, the dosage administered is between about 1 mg/kg to about 3 mg/kg of the patient's body weight.

The illudofulvene can be administered by any convenient route, for example by infusion or bolus injection. Administration can be systemic or local. Various delivery systems are known, e.g., encapsulation in liposomes, micro-particles, microcapsules, capsules, etc., and can be used to administer an illudofulvene. In certain embodiments, more than one illudofulvene can be administered to a patient.

In specific embodiments, it can be desirable to administer one or more illudofulvene locally to the area in need of treatment. This can be achieved, for example, and not by way of limitation, by local infusion during surgery; topical application, e.g., in conjunction with a wound dressing after surgery; by injection; by means of a catheter; or by means of an implant, the implant being of a porous, non-porous, or gelatinous material, including membranes, such as sialastic membranes, or fibers. In one embodiment, administration can be by direct injection at the site (or former site) of a cancer, tumor or neoplastic or pre-neoplastic tissue. In another embodiment, administration can be by direct injection at the site (or former site) of a manifestation of an autoimmune disease.

In yet another embodiment, the illudofulvene can be delivered in a controlled release system, such as but not limited to, a pump or various polymeric materials can be used. In yet another embodiment, a controlled-release system can be placed in proximity of the target of the illudofulvene, e.g., the liver, thus requiring only a fraction of the systemic dose.

The present compositions can take the form of solutions, pellets, powders, sustained-release formulations, or any other form suitable for use.

In an embodiment, the illudofulvenes are formulated in accordance with routine procedures as a pharmaceutical composition adapted for intravenous administration to animals, particularly human beings. Typically, the carriers or vehicles for intravenous administration are sterile isotonic aqueous buffer solutions. Where necessary, the compositions can also include a solubilizing agent. Compositions for intravenous administration can optionally comprise a local anesthetic such as lignocaine to ease pain at the site of the injection. Generally, the ingredients are supplied either separately or mixed together in unit dosage form, for example, as a dry lyophilized powder or water free concentrate in a hermetically sealed container such as an ampoule or sachet indicating the quantity of active agent. Where an illudofulvene is to be administered by infusion, it can be dispensed, for example, with an infusion bottle containing sterile pharmaceutical grade water or saline. Where the illudofulvene can be administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients can be mixed prior to administration.

The composition can include various materials that modify the physical form of a solid or liquid dosage unit. For example, the composition can include materials that form a coating shell around the active ingredients. The materials that form the coating shell are typically inert, and can be selected from, for example, sugar, shellac, and other enteric coating agents. Alternatively, the active ingredients can be encased in a gelatin capsule.

Whether in solid or liquid form, the present compositions can include a pharmacological agent used in the treatment of cancer, an autoimmune disease or an infectious disease.

The illudofulvene can also be used in an in vitro or ex vivo fashion, such as for the treatment of certain cancers, including, but not limited to leukemia and lymphomas, such treatment involving autologous stem cell transplants. This can involve a multi-step process in which the animal's autologous hematopoietic stem cells are harvested and purged of all cancer cells, the animal's remaining bone-marrow cell population can then eradicated via the administration of a high dose of an illudofulvenes with or without accompanying high dose radiation therapy, and the stem cell graft can be infused back into the animal. Supportive care can then be provided while bone marrow function is restored and the patient recovers.

A method of ablating cancer cells comprising the steps of selecting an illudofulvene which is cytotoxic to the cancer cell and treating the cancer cells with the illudofulvene.

In an embodiment of the present invention, a tumor sample from a patient can be harvested and processed to identify the relative expression of genes encoding the biomarkers described in this application that predict the responsiveness of the patient to Illudofulvenes. Once the relative expression of the biomarkers has been determined, the responsiveness of the cancer patient to treatment with Illudofulvenes, can be determined as outlined in the FIG. 5.

In an embodiment of the present invention, a cancer patient who has an upregulated expression of either PTGR1 or PTGR2 will display responsiveness to treatment with an Illudofulvene if there is a deficiency in any one (or more) of the TCR genes. In an embodiment of the present invention, a cancer patient who has an upregulated expression of either PTGR1 or PTGR2 or an upregulated expression of Myc will display responsiveness to treatment with an Illudofulvene if there is a deficiency in any one (or more) of the TCR genes.

In an embodiment of the present invention, a cancer patient who has an upregulated expression of Myc and does not have an upregulated expression either PTGR1 or PTGR2 will display responsiveness to treatment with an Illudofulvene if there is a deficiency in any two (or more) of the TCR genes. In an embodiment of the present invention, a cancer patient who does not have upregulated expression of PTGR1 and PTGR2 but has an upregulated expression of Myc will display responsiveness to treatment with an Illudofulvene if there is a deficiency in any two (or more) of the TCR genes.

In an embodiment of the present invention, a cancer patient who does not have an upregulated expression of PTGR1 and PTGR2 and does not have an upregulated expression of Myc will display responsiveness to treatment with an Illudofulvene if there is a deficiency in any three (or more) of the TCR genes. In an embodiment of the present invention, a cancer patient who does not have an upregulated expression of Myc and does not have an upregulated expression of PTGR1 and PTGR2 will display responsiveness to treatment with an Illudofulvene if there is a deficiency in any three (or more) of the TCR genes. In an embodiment of the present invention, a cancer patient who does not have an upregulated expression of PTGR1, PTGR2, and Myc will display responsiveness to treatment with an Illudofulvene if there is a deficiency in any three (or more) of the TCR genes.

Quantification of Gene mRNA Expression by Probe Analysis.

Tumor tissue can be harvested and total RNA can be purified using Trizol Reagent (Invitrogen Life Technology, 15596-018, Waltham. MA) and the RNeasy Mini Kit (Qiagen, 74104, Germantown, MD). The mRNA can then be purified (from the total RNA) using an Oliogotex mRNA kit (Qiagen, 70022, Germantown, MD). The isolated tissue mRNA can then be fixed to a nylon membrane (ThermoFisher, AM10102, Waltham. MA), by UV crosslinking (Stratagene, UC Crosslinker 2400, La Jolla, CA), for subsequent analysis of the relative quantity expressed by a specific gene. Probes can then be prepared by PCR labeling. Probes can be labeled with a radioactive isotope, e.g. P32 or P33, but the preferred approach is use of a nonradioactive label such as Digoxigen-11-dUTP (Sigma Aldrich, 1157315290, St Louis, MO). Other potential nonradioactive probe labeling methods include the use of Biotin-16-UTP, Biotin-11-CTP or Fluoroscein-12-UTP. In PCR labeling, a thermostable polymerase (Agilent, 600195, Santa Clara, CA) incorporates the Digoxigen-11-dUTP as it amplifies a specific region of template DNA that is often contained in a plasmid. The specificity of the probe (or gene under study) can be determined by the two PCR primers utilized. The result can be a highly labeled, highly specific and sensitive hybridization probe for quantifying the amount of corresponding mRNA in a tissue sample. The hybridization of the probe to the membrane can be performed normally in roller bottles in a hybridization oven, followed by high stringency washes at elevated temperatures up to 70° C., to remove nonspecific binding and background signal. The digoxigen labeled probes are detected using labeled anti-digoxigen antibody (Abcam product number 420) or Fab fragments (Sigma Aldrich, 11207733910, St Louis, MO). If another label is utilized for probe labeling, such as biotin, then an anti-biotin labeled antibody can be used. Visualization and quantification of signal can be performed using a phosphoimager (Storm, 860 imager, Ramsey, MN) or a fluoroimager, depending upon the type of label used in the detection antibody.

In an embodiment of the invention, the tumor tissue is harvested and mRNA purified from the tumor. The mRNA is processed, subjected to electrophoresis and then analyzed with the desired probe for individual gene products, as well as for the three “housekeeping” genes (ACTB, TFRC, and GAPD). The required probes specific for detecting a gene product are prepared using PCR with the stated primers specific for that gene (Table 1) and labeled with a detection product as described in this application. The raw expression of each gene product is then determined using a densitometer appropriate for the label used (fluorescent, visible, photometer, radioisotope, etc.) Next a “relative ratio of expression” is determined for each of the gene products under investigation versus the individual “housekeeping genes” as described above in the Microarray section. These ratios are then compared to ratios obtained for a cell known to be sensitive to an Illudofulvene as well as to ratios for a cell know to be resistant to an Illudofulvene. One can then determine whether (i) a PTGR gene is upregulated, and/or (ii) the MYC gene is upregulated and (iii) there is a deficiency in a specific TCR gene. The tumor can then be classified as either sensitive, or resistant, to Illudofulvenes based on this analysis.

TABLE 1

Primers for use with gene mRNA expression,

according to embodiments of the invention.

Upper
Lower

Primer

Primer
Primer

Annealing

SEQ ID
SEQ ID
Probe
Temp

GENE
NOS:
NOS:
(bp)
(° C.)

ERCC1
275
276
467
59

ERCC2
277
278
601
60

ERCC3
279
280
415
59

ERCC4 (set 1)
281
282
1217
56

ERCC4 (set 2)
283
284
500
58

ERRC5
285
286
872
56

ERCC6
287
288
876
55

ERCC6L
289
290
629
62

ERCC6L2
291
292
594
63

ERCC8
293
294
534
50

GTF2H1
295
296
1139
66

GTF2H2
297
298
802
64

GTF2H3
299
300
344
62

GTF2H4
301
302
560
62

GTF2H5
303
304
606
63

MYC
305
306
630
63

PTGR1
307
308
621
61

PTGR2
309
310
626
63

RAD18
311
312
601
58

USP7
313
314
600
62

UVSSA
315
316
595
62

XPA
317
318
459
64

XPC
319
320
855
62

XPE
321
322
629
62

Controls

ACTB β-Actin
323
324
566
59

TFRC
325
326
1096
53

GAPD
327
328
547
60

The cDNA genes under investigation were isolated internally (by applicant) from a lambda gt11 expression vector using primers above, then cloned into a plasmid. All sequences were confirmed by nucleotide sequencing. The genes at the time were not available from commercial sources.

Microarray Analysis.

Next an RNase H-mediated second-strand cDNA synthesis can be performed, the double-stranded cDNA can be purified, and this double-strand cDNA serves as the template in a subsequent in vitro transcription (IVT) reaction (Affymetrix GeneChip IVT kit 900688, Waltham. MA). The IVT reaction can be carried out in the presence of provided T7 RNA Polymerase and a biotinylated nucleotide analog/ribonucleotide mix for complementary RNA (cRNA) amplification and biotin labeling. The biotinylated cRNA targets are then cleaned up and fragmented using the buffer provided in the kit.

Next the cRNA under study and a control oligonucleotide are prepared in a hybridization solution (Affymetrix, 900454, Waltham. MA) and added to the Affymetrix HG-U133 Plus 2 array cartridge, then placed in the hybridization oven at 45 C.° with rotation at 60 RPM, for 16 hours. The HG-U133 cartridge can be then washed and stained using the appropriate Fluidics station per manufacturer's protocol. The array can then be scanned using an appropriate device such as the Affymetrix GeneChip scanner or a generic scanner such as the Agilent GeneArray scanner. Normalization and analysis of data can be accomplished using the NetAffx Software (Thermo Fisher, Waltham. MA), the Gene Spring GX 9 software (Agilent, Santa Clara, CA) or open source academic software such as R/Bioconductor.

The specific probe sequences and corresponding Affymetrix IDs, for the biomarkers involved, for the Human Genome U133 Plus 2.0 array (HG-U133 Plus 2) are provided in Tables 2 and 3.

In an embodiment of the invention, the tumor tissue is harvested and the relative expression of the individual gene products, as well as the mRNA expression for the three “housekeeping” genes (ACTB, TFRC, GAPD), is quantified by a microarray apparatus using the specific probes for each gene product (see Tables 2 and 3). Next a “relative ratio of expression” is determined for each of the gene products under s investigation versus the individual “housekeeping genes” (e.g. ERCC6 expression/ACTB expression). This is accomplished by dividing the raw value of expression for each gene product under study by the raw value of expression for the desired “housekeeping gene”. As an example the ERCC6/ACTB, ERCC6/TFRC, and the ERRC6/GAPD expression ratios for the tumor under study are calculated. Next an analysis is performed by comparing each relative ratio for each gene using each housekeeping gene, and comparing the ratio to the same ratio for a cell known to be sensitive to an Illudofulvene as well as a cell know to be resistant to an Illudofulvene. As an example the ERCC6/ACTB, ERCC6/TFRC, and the ERRC6/GAPD expression ratios for the tumor under study are compared to the same ratios in a cell known to be sensitive to an Illudofulvene as well as a cell know to be resistant to an Illudofulvene. Note that only one of the many gene ratios could be similar to a ratio derived from a cell known to be sensitive to an Illudofulvene in order to classify the tumor as TCR deficient. If any single component of the TCR pathway is deficient then that single deficiency is sufficient to render a tumor sensitive to Illudofulvenes (see Table 10). Similarly, the ratios for the individual expression of PTGR1, PTGR2, and MYC to the three “housekeeping” genes can be derived and each ratio can be compared to the same ratio in a cell known to be sensitive to an Illudofulvene as well as a cell know to be resistant to an Illudofulvene. This allows a classification as to whether PTGR1, PTGR2, or MYC are either upregulated or not upregulated and which specific gene(s) are upregulated for that individual tumor. The classification also allows determination of the specific TCR genes that are deficient.

TABLE 2

Probe Sequences to analyze not upregulated gene

products that confer sensitivity to Illudofulvenes

Gene
Affymetrix ID
SEQ ID NOS:

ERCC8
201562_at
35-45

ERCC6
207347_at
46-56

GTF2H1
202451_at
57-67

GTF2H2
230177_at
68-78

GTF2H3
239346_at
79-89

GTF2H4
203577_at
90-100

GTF2H5
213357_at
101-111

USP7
2014498_at
112-122

UVSSA
241201_at
123-133

ERCC3
202176_at
134-144

ERCC2
213468_at
145-155

ERCC4
210158_at
156-166

ERCC5
202414_at
167-177

XPA
205672_at
178-188

RAD18
223417_at
189-199

ERCC6/L
219650_at
200-210

ERCC6/L2
228211_at
211-221

ERCC1
203719_t
238-248

Affymetrix IDs refer to the array type Human Genome U133 Plus 2.0 (HG-U133 Plus 2)

TABLE 3

Probe Sequences to analyze upregulated gene products

that confer sensitivity to Illudofulvenes

Gene
Affymetrix ID
SEQ ID NOS:

MYC
20243_s_at
224-235

PTGR1
231897_at
251-261

PTGR2
230774_at
264-274

Affymetrix IDs refer to the array type Human Genome U133 Plus 2.0 (HG-U133 Plus 2)

RT-PCR Analysis.

In an embodiment of the invention, the RT-PCR Analysis can be carried out using the Hydrolysis Taqman Fluorescent probe approach. Tumor tissue can be harvested and total RNA can be purified using Trizol Reagent (Invitrogen Life Technology, 15596-018, Waltham. MA) and the RNeasy Mini Kit (Qiagen, 74104, Germantown, MD). The mRNA can then be purified (from the total RNA) using an Oliogotex mRNA kit (Qiagen, 70022, Germantown, MD). Then reverse transcription can be performed using oligo(dT) and random hexamers and SuperScript® II (Thermofisher, #18064014, Waltham, MA) to produce cDNA. Each reaction mixture contains approximately 50 ng of tumor-derived cDNA.

The desired gene primers, and associated hydrolysis oligonucleotide probe (see Table 4) with a 5′ fluorescent reporter and a 3′ quencher, are obtained from Integrated DNA Technologies. RT-PCR reaction conditions are an initial denaturation for 2 minutes at 95° C. and 45 cycles of 94° C. for 15 seconds, annealing at 56° C. for 15 seconds and 72° C. for 30 seconds, with final extension at 72° C. for 10 minutes. The hydrolysis probe for the gene under study can be labeled at the 5′ end with the fluorophore FAM (fluorescein). A control gene can be amplified as an internal control to check the specificity of the reaction. The hydrolysis probe can be labeled at the 5′ end with the fluorophore HEX (555) to allow for differentiation of fluorescence arising from the biomarker hydrolysis probe and the internal control probe. All RT-PCR assays are run in triplicate using TaqMan® Gene Expression Master Mix (ThermoFisher. #4369016, Waltham. MA).

Quantitative results are obtained by monitoring the increase in fluorescence (which occurs upon release of the fluorescent reporter molecule from the hydrolysis probe and the associated quencher) and can be proportional to the amount of amplified product produced, using a Roche Light cycle Model 2.0 (or later version).

TABLE 4

Required RT-PCR primers and hydrolysis probe for each Biomarker.

Upper
Hydrolysis
Lower

Primer
Probe
Primer

SEQ ID
SEQ ID
SEQ ID
Probe
Temp

GENE
NOS:
NOS:
NOS:
(bp)
(° C.)

ERCC1
852
853
854
139
61

ERCC2
855
856
857
141
60

ERCC3
858
859
860
143
60

ERCC4
861
862
863
137
60

ERRC5
864
865
866
143
60

ERCC6
867
868
869
148
60

ERCC6L
870
871
872
137
61

ERCC6L2
873
874
875
148
60

ERCC8
876
877
878
142
60

GTF2H1
879
880
881
89
61

GTF2H2
882
883
884
146
60

GTF2H3
885
886
887
141
61

GTF2H4
888
889
890
150
61

GTF2H5
891
892
893
149
61

MYC
894
895
896
108
60

PTGR1
897
898
899
150
60

PTGR2
900
901
902
148
60

RAD18
903
904
905
137
60

USP7
906
907
908
135
60

UVSSA
909
910
911
114
61

XPA
912
913
914
140
60

XPC
915
916
917
144
61

ACTB β-Actin
918
919
920
148
61

(control)

TFRC (control)
921
922
923
127
60

GAPD (control)
924
925
926
143
60

TCR Tumor Mutation Panel (22 Genes) Analysis.

The TCR Tumor Mutation Panel targets a total of 22 clinically relevant genes. Specifically, the panel interrogates all exons of 22 genes known to be associated with sensitivity to Illudofulvenes.

The TCR Tumor Mutation Panel (see Table 5) is a highly multiplexed targeted re-sequencing assay for detecting somatic variants across key genes in cancer genomes [see (i) Illumina HiSeq® 2500 System User Guide. Illumina Proprietary Part #15035786 Rev. D, November 2014; (ii) HiSeq Rapid SBS Kit v2 Reagent Prep Guide. Illumina Proprietary Part #15058772 Rev. A, November 2014; (iii) HiSeq Rapid Cluster Kit v2 Reagent Prep Guide. Illumina Proprietary Part #15059131 Rev. A, November 2014; (iv) Sequencing Analysis Viewer Software User Guide. Illumina Proprietary Part #15020619 Rev. F, October 2014; of which (i)-(iv) are herein expressly incorporated by reference in their entireties for all purposes]. The analysis entails hybridization-based capture of 513 probes (custom mix synthesis from Integrative DNA Technologies) covering 47,848 base pairs that encompass the 281 exons and regulatory regions of these 22 genes. The generation of a library of multiplexed, enriched genomic regions can be followed by a sequencing-by-synthesis assay using massively parallel 100 base pair sequence reads. Once data are generated, a series of bioinformatics tools are used to align the reads to the genome, call variants, and annotate variants with respect to somatic status and functional predictions. This analysis can detect single nucleotide variants, short insertion/deletions, and splice variants predicted to be deleterious. In addition, the analysis can detect a set of targeted fusions or rearrangement.

Genomic DNA extracted from tumor tissue can be sheared, end repaired, A-tailed, adapter ligated, amplified, hybridized to the capture (bait) library primers, then re-amplified per the HiSeq Reagent Pre Guides (Illumina #15058772 and #15059131). The subsequent sample pool can then be sequenced to a depth of 300× on an Illumina HiSeq 2500 workstation per the user guide (Illumina #15035786). The primary data analysis can be accomplished by the on-board software (Real Time Analysis). Secondary analysis can be accomplished by uploading base call files (*.bcl) to a genomics server designed to generate reads and read alignments, followed by comparison to the DNA sequence for the 22 genes as listed in the latest version of the Genomic Reference Consortium Human (current build 37). This allows identification of tumors that are deficient in TCR capacity, on the basis of a deficiency in one (or more) of the components of the TCR pathways, and which can be responsive to treatment with an Illudofulvene. In addition, tumors that are deficient in PTGR metabolic activity (either PTGR1 or PTGR2) can also be identified, and such tumors excluded from treatment based on the expected lack of response to therapy.

In an embodiment of the invention, a tumor sample is obtained from a patient, prepared, and the genomic DNA isolated and processed as described in this application. The TCR 22 gene mutation panel (Table 5) is then obtained by sequencing in entirety these genes, and results analyzed versus the latest version of the Genomic Reference Consortium Human (current build 37, Table 6), as described in this application. Tumors deficient in TCR capacity, on the basis of a deficiency in one (or more) of the components of the TCR pathways due to a mutation, and which can be responsive to treatment with an Illudofulvene, can then be identified. Tumors that are deficient in PTGR metabolic activity (either PTGR1 or PTGR2) can also be identified, and such tumors excluded from treatment based on the expected lack of response to therapy, subject to the flow chart of FIG. 5. It can also be determined whether MYC is upregulated or downregulated.

TABLE 5

The twenty two (22) Gene

TCR Tumor Mutation Panel

GENE
SEQ ID NOS:

ERCC1
329-343

ERCC2
344-374

ERCC3
375-402

ERCC4
403-429

ERCC5
430-440 and 451-475

ERCC6
476-539

ERCC6L
540-571

ERCC6L2
572-596

GTF2H1
597-615

GTF2H2
616-631

GTF2H3
632-646

GTF2H4
647-665

GTF2H5
666-668

MYC
669-681

PTGR1
682-696

RAD18
697-714

PTGR2
715-727

XPA
728-736

XPC
737-777

USP7
768-809

UVSSA
810-835

ERCC8
836-851

TABLE 6

A list of genes comprising components of the TCR-NER pathway, post-

replication repair pathway, or bio activation of Illudofulvenes of which an

alteration of one or more of the components will confer sensitivity to

illudofulvenes, according to various embodiments of the invention.

Gene (HGNC
Gene
Representative
SEQ ID
Representative
SEQ ID

Symbol)
Aliases
mRNA Sequence
NO
Protein Sequence
NO

ERCC8
CSA,
NM_001290285
1
NP_000073
2

CKN1,

UVSS2

ERCC6
CSB,
NM_001277058.1
3
NP_000115.1
4

ARMD5,

CKN2,

COFS,

COFS1,

RAD26

POF11,

UVSS1

GTF2141
P62,
NM_001142307.1
5
NP_005307.1
6

BTF2P62,

TUB1,

TFIIH

SUBUNIT 1

GTF2H2
BTF2P44. P44
NM_001515.3
7
NP_001506.1
8

GTF2H3
P34, TFB4
NM_001516.4
9
NP_001507.2
10

GTF2H4
P52, TFB2
NM_001517.4
11
NP_001508.1
12

GTF2H5
TTDA,
NM_207118.2
13
NP_997001.1
14

TFB5,

TGF2H5,

TTD3,

C6orf175,

bA120J8.2

USP7
HAUSP,
NM_003470.3
15
NP_0013461.2
16

TEF1

UVSSA
KIAA1530,
NM_020894.3
17
NP_001304863.1
18

UVSS3

ERCC3
XPB,
NM_001303418.1
19
NP_001290347.1
20

RAD25,

TTD2,

BFT2

ERCC2
XPD,
NM_000400.3
21
NP_000391.1
22

COFS2,

EM9,

TTD1

ERCC4
XPF,
NM_005236.2
23
NP_005227.1
24

ERCC11,

FANCQ,

RAD1,

XFEPS

ERCC5
XPG,
NM_000123.3
25
NP_000114.2
26

COFS3,

ERCM2,

UVDR,

XPGC

XPA
XP1,
NM_001354975.1
27
NP_000371.1
29

XPAC

RAD18
RNF73
NM_020165.3
29
NP_064550.3
30

ERCC6L
PICH,
NM_017669.3
31
NP_060139.2
32

RAD26L

ERCC6/L2
BMFS2,
NM_020207.4
33
NP_064592.2
34

C9orf102,

RAD26L,

SR278

MYC
v-myc,
NM_002467.5
222
NP_002458
223

c-myc,

bHLHe39,

MYCC

ERCC1
COFS4,
NM_202001.2
236
NP_973730.1
237

UV20,

RAD10

PTGR1
PGR1,
NM_001146109.1
249
NP_001139581
250

PRG-1,

LTBD4H,

ZADH3

PTGR2
PGR2,
NM_001146154.1
262
NP_001139626.1
263

ZADH1,

HEL-S-298

TABLE 7

A list of biomarkers that when downregulated (expression

decreased) confer sensitivity to illudofulvenes, according

to various embodiments of the invention.

Gene
Affymetrix ID
SEQ ID NO

ERCC8
201562_at
35-45

ERCC6
207347_at
46-56

GTF2H1
202451_at
57-67

GTF2H2
230177_at
68-78

GTF2H3
239346_at
79-89

GTF2H4
203577_at
90-100

GTF2H5
213357_at
101-111

USP7
2014498_at
112-122

UVSSA
241201_at
123-133

ERCC3
202176_at
134-144

ERCC2
213468_at
145-155

ERCC4
210158_at
156-166

ERCC5
202414_at
167-177

XPA
205672_at
178-188

RAD18
223417_at
189-199

ERCC6/L
219650_at
200-210

ERCC6/L2
228211_at
211-221

ERCC1
203719_t
238-248

Affymetrix IDs refer to the array type Human Genome U133 Plus 2.0 (HG-U133 Plus 2)

TABLE 8

A list of biomarkers that when upregulated (expression

increased) confer sensitivity to illudofulvenes,

according to various embodiments of the invention.

Gene
Affymetrix ID
SEQ ID NO

MYC
20243_s_at
224-235

PTGR1
231897_at
251-261

PTGR2
230774_at
264-274

Affymetrix IDs refer to the array type Human Genome U133 Plus 2.0 (HG-U133 Plus 2).

TABLE 9

The toxicity of Illudofulven Analog 001/Analog 002

to either 2 hour or 48 hour incubation, according

to various embodiments of the invention.

Cytotoxicity?

Tumor cell line
2 hr incubation
48 hr incubation

Myeloid leukemias
Yes
Yes

Breast carcinomas
Yes
Yes

Epidermoid
Yes
Yes

Ovarian
Yes
Yes

Lung carcinomas
Yes
Yes

Prostate carcinomas
Yes
Yes

B cell leukemias
No
Yes

T cell leukemias
No
Yes

Fibroblasts (normal)
No
Yes

TABLE 10

A comparison of the DNA damage profile of Illudofulven

Analog 001/Analog 002 with Other Drugs and UV

irradiation, according to embodiments of the invention.

Other

Illudofulvene

Gene
Drugs*
UV
Analog 001/002

XP-A
+
+
+

XP-B
0
+
+

XP-C
+
+
0

XP-D
0
+
+

XP-E
+
+
0

XP-F
+
+
+

CS-A
+/−
+
++

CS-B
+/−
+
++

ERCC1
+
+
+

ERCC5
+
+
+

*as defined herein.

TABLE 11

The multidrug resistance of Illudofulvene

Analog 001/Analog 002, according to

various embodiment of the invention.

Resistance to

Mechanism
Illudofulvene

of Multi-drug
Analog 001/

Resistance
Analog 002

Gp170/MDR1
No

Gp180/MRP
No

Topoisomerase I
No

Topoisomerase II
No

MVP/LRP (vault)
No

Thiol content/GST pi
No

DNA repair
No

Myc expression
No

Bcl-2 expression
No

BRCA status
No

P53 status
No

P21 status
No

MGMT expression
No

Microtubulin alteration
No

TABLE 12

The cytotoxic IC₅₀values (micromolar, 2 hour exposure, N = 3, mean ± SD)

for illudin M, llludofulvene Analog 108 and Illudolulvene Analog 110

for cells expressing the estrogen receptor (ER) (MCF7)and cells not

expressing the ER (HT29), according to embodiments of the invention.

Analog
HT29 (ER Negative)
MCF7 (ER positive)

Illudin M
0.52 ± 0.10
0.48 ± 0.13

108
>55
14.1 ± 2.8

110
>19
2.0 ± 0.1

TABLE 13

The ability of Illudofulvene analogs to inhibit tumor cell growth, according

to various embodiments of the invention.

Mean IC50 value (nM) ± SD, N = 3 unless otherwise indicated

Illudofulvene
MV522 Target Cell Line
8392B Nontarget Cell Line

Analog
2 hr exposure
48 hr exposure
2 hr exposure
48 hr exposure

001
2200 ± 100
350 ± 20

830 ± 100

002
110 ± 40
70 ± 10
26000 ± 4500
800 ± 100

004
4200
600

008
870 ± 90
630 ± 80
12200 ± 700
15100 ± 2200

009
500 ± 30
850 ± 180
47100 ± 11000
43200 ± 2300

010
8900 ± 1500
170 ± 60
29400 ± 1600
14500 ± 1700

011
4900 ± 900
1200 (N = 2)
>100000
40400 ± 6700

012
5150 ± 1350
320 ± 90
42200 ± 5000
18800 ± 2800

013
5100 ± 700
270 ± 130
11900 ± 1300
4200 ± 400

014
115 ± 30
460 ± 120
9650 ± 200
1100 ± 300

015
1800 ± 200
480 ± 110
810 ± 260
1300 ± 150

016
490 ± 130
440 ± 90
>100000
870 ± 60

017
2400 ± 360
320 ± 60
14700 ± 900

018
8800 ± 2900

4200 ± 1300

019
470 ± 60
660 ± 80
>75000

020
530 ± 140
230 ± 10
25000 ± 3100

021
2400 ± 1000
930 ± 250
34400 ± 9400

022
700 ± 200
680 ± 180
31700 ± 1400

023
2900 ± 1140
2750 ± 500
>138000

024
1800 ± 200
1200 ± 300
12800 ± 2100

025
1300 ± 310
1200 ± 100
>25000

030

>3000

031

>3000

032
600 ± 190
210 ± 30
>30000

033
10000 ± 1100
4600 ± 200
29900 ± 3300

034
1400 ± 170
490 ± 40
>100000
4400 ± 200

035
5600 ± 600

>150000

037
26000 ± 5000
29200 ± 2300
>85000

038
750 ± 60

24900 ± 8000

039
1500 ± 240
600 ± 40
24600 ± 2400
820 ± 250

040
3400 ± 360
700 ± 90
24000 ± 3300
5200 ± 470

060
19400 ± 1800

27600 ± 3000

062
2600 ± 300
660 ± 200
37100 ± 2300

063
43000 ± 5700
580 ± 250

064
28000 ± 4600
1200 ± 300

065
6200 ± 1100
2500 ± 1200

075
19600 ± 9700

62000 ± 3600

076
24000 ± 6100

39500 ± 7200

077
9200 ± 1200

078
20400 ± 6300

>100000

079
7700 ± 3500

>100000

080
8800 ± 2400

>100000

081
>80000

>80000

082
50600 ± 7100

>100000

083

37200 ± 2900
>42000

084

28200 ± 1400
>42000

085
>40000

>40000

087
>40000
24700 ± 3900
>40000

089
19300 ± 5700
15500 ± 2800
>60000

090
2500 ± 400
2900 ± 400
1600 ± 200
3800 ± 300

094
800 ± 100
210 ± 20
9000 ± 1700
110 ± 10

096
2700 ± 400
6200 ± 600
>88000
>3000

097
2900 ± 100

>82000

098
18800 ± 2500
4600 ± 250
>65000
11700 ± 1800

099
8400 ± 1100
1800 ± 200
4000 ± 400
300 ± 20

100
>10000
1700 ± 500

101
>8000
>7500

102
>13000
1300 ± 100

103
31800 ± 4900
5900 ± 400
12100 ± 2000
2300 ± 200

104
6300 ± 400
6000 ± 500
36400 ± 6500
2700 ± 600

105
7300 ± 1200
2100 ± 400
>100000

106
5200 ± 1000

>83000

107
>50000
1600 ± 100
>50000

108
12300 ± 2300
520 ± 50
>55000
6000 ± 1600

109
>50000

>50000

110
>55000
1400 ± 100
>55000
25300 ± 2100

111
16700 ± 2100
11900 ± 2800
34600 ± 2100
10200 ± 1000

112
10000 ± 2000
6700 ± 1200
14900 ± 100
5200 ± 300

113
85000 ± 700
14100 ± 3000
>93000
7800 ± 1000

114
1500 ± 100
260 ± 70
25100 ± 1000
700 ± 100

115
1500 ± 100
70 ± 5
1600 ± 700
630 ± 60

116
400 ± 100
1000 ± 50
7000 ± 400
170 ± 30

117
1100 ± 100
100 ± 30
7900 ± 1600
10 ± 2

118
14000 ± 2000
740 ± 120
24500 ± 4500
2000 ± 400

119
1100 ± 70
270 ± 40
>33000
>10000

120
2800 ± 900
600 ± 100
19100 ± 4600
510 ± 110

121
300 ± 10
90 ± 10
15200 ± 6000
1300 ± 500

122
6400 ± 300
2400 ± 300
14500 ± 1200
1100 ± 300

123
1900 ± 400
600 ± 60
450 ± 30
2400 ± 500

124
2800 ± 700
870 ± 350
>30000
2400 ± 550

125
3700 ± 600
1200 ± 200
15500 ± 1400
600 ± 100

126
2100 ± 500
900 ± 100
>30000
330 ± 80

127
870 ± 30
340 ± 90
>30000
100 ± 40

128
840 ± 230
370 ± 50
>35000
800 ± 70

129
>136000
19700 ± 1900
>136000
39400 ± 9200

130
700 ± 100
130 ± 40
27,000 ± 7000
4400 ± 500

133
58800 ± 6600
15800 ± 2600
12200 ± 2300
2700 ± 400

134
50000 ± 6000
28000 ± 4000
43900 ± 5100
8500 ± 2000

135
1600 ± 300
22 ± 4
70 ± 20
22 ± 2

136
430 ± 10
130 ± 10
>6200
25 ± 2

137
850 ± 110
1200 ± 100
8500 ± 1200
710 ± 60

138
2100 ± 200
1000 ± 200
5400 ± 200
820 ± 230

139
6400 ± 900
3400 ± 500
11600 ± 900
2600 ± 1000

140
17100 ± 5100
>14000
12700 ± 300
>14000

141
11400 ± 1000
3700 ± 800
13700 ± 1900
1100 ± 140

142
90 ± 10
24 ± 7
6400 ± 1100
80 ± 6

143
43500 ± 11300
11400 ± 1800
56500 ± 20000
3600 ± 700

146
2500 ± 400
740 ± 280
13,000 ± 1200

147
>76000
26100 ± 12900
>76000
43800 ± 3000

148
17100 ± 1100
6800 ± 1100
61000 ± 11600
6700 ± 1600

149
2900 ± 1000
1500 500
44600 ± 1400
4100 ± 900

150
9500 ± 1600
1400 ± 400
59000 ± 5500
10600 ± 800

151
7900 ± 400
4200 ± 1600
25500 ± 1200
6600 ± 2300

152

6400 ± 1200
49000 ± 7700
9100 ± 100

153
8700 ± 2700
10900 ± 3400
>90000
15800 ± 9600

154
>70000
61300 ± 10000
>70000
46,700 ± 13100

155
8200 ± 1200
3600 ± 400
17,000 ± 4000
9100 ± 1100

156
7200 ± 500
3100 ± 100
32,300 ± 9,400
5500 ± 1200

157
>400,000
>123,000
>350,000
13100 ± 1600

158
>175,000
>175,000
>200,000
61,000 ± 9,000

159
2700 ± 400
120 ± 10
13,700 ± 4,200
<10 nM

160
1900 ± 200
500 ± 200
52,400 ± 17,800
3200 ± 1100

161
2800 ± 500
3300 ± 700
13,800 ± 3,400
>10,000

163
3500 ± 800
820 ± 40
18600 ± 800
910 ± 100

164

70 ± 10
3500 ± 1600
130 ± 40

165
7700 ± 1100
290 ± 40
11000 ± 3300
11000 ± 1000

166
6500 ± 600
7200 ± 1900
6500 ± 2100
6000 ± 1500

167
14800 ± 2200

18500 ± 2300

169
7100 ± 600

2300 ± 600

177
7500 ± 800
1900 ± 800
73000 ± 5000
4100 ± 1300

178
21000 ± 4000
1000 ± 100
32000 ± 9000
>8000

180
19900 ± 300
>4000
5200 ± 1800
660 ± 50

182
99000 ± 12000
38000 ± 8200
39000 ± 7000
18700 ± 2700

183
>120,000
>275,000
>120,000
>235,000

184
800 ± 300
210 ± 20
>100,000
>10000

185
1700 ± 600
1900 ± 100

186
144000 ± 32000
70000 ± 16000
79000 ± 24000
48000 ± 2000

187
1300 ± 400
900 ± 200
3200 ± 800
3200 ± 700

189
8900 ± 2500
6100 ± 2600
41,000 ± 3700

190
19,000 ± 4000
>9,000
56,000 ± 2000
>9,000

191
>140,000
49,000 ± 13000
>140,000
15000 ± 4000

192
1,600 ± 200
700 ± 100
8700 ± 1700
200 ± 30

193
1400 ± 400
2500 ± 600
48,000 ± 7000
>11,000

195
1400 ± 200
390 ± 120
21,000 ± 6000
4300 ± 1200

196
840 ± 100
450 ± 120
80,000 ± 5000
>9,200

197
950 ± 70
500 ± 100
9500 ± 400
[1,300 ± 100

198
700 ± 100
2800 ± 600
>8,200
>82,000

199
4700 ± 600
2500 ± 1100
>93,000
>9,300

201
360 ± 110
260 ± 70
13,000 ± 1700
26,000 ± 7000

202
1200 ± 100
650 ± 100
>62,000
>6200

203
760 ± 170
940 ± 330
48,000 ± 6000
>5500

204
220 ± 40
1600 ± 300
4100 ± 800
8600 ± 800

205
8400 ± 2200
1200 ± 400
>185,000
>2,600

206
610 ± 40
230 ± 20
20,000 ± 1000
8200 ± 200

207
570 ± 60
410 ± 60

208
1200 ± 100
930 ± 160
25,000 ± 3000

209
3900 ± 1100
610 ± 100
>90,000

210
40,000 ± 4000
5500 ± 600

211
470 ± 120
430 ± 100
59,000 ± 9000

212
80 ± 10
55 ± 5

213
2300 ± 700
1700 ± 700

214
2900 ± 800
360 ± 30

215
26,000 ± 3000
490 ± 120

216
460 ± 60
150 ± 40

217
2,200 ± 100
2,200 ± 100
43,000 ± 4,000
>7,000

218
10,000 ± 3,000
600 ± 200
15,000 ± 6,000
600 ± 100

219
>52,000
>52,00
>52,000
>52,000

220
90 ± 10
130 ± 10
101,000 ± 18,000
40,000 ± 3,000

221
>21,000
2,500 ± 200
>21,000
>21,000

222
5,000 ± 100
1,100 ± 100
9,300 ± 200
330 ± 60

223
20,000 ± 3,700
2,700 ± 300
>185,000
>55,000

224
>200,000
>130,000
>200,000
>130,000

225
47,000 ± 4,000
55,000 ± 11,000
>350,000
33,000 ± 13,000

226
>59,000
>59,000
>59,000
>59,000

227
>57,000
4,400 ± 700
>57,000
16,000 ± 4,000

228
>38,000
>38,000
24,000 ± 3,000
>38,000

229
>56,000
>2,000
>56,000
>2,000

230
620 ± 80
100 ± 10
38,000 ± 5,000
1,000 ± 200

231
1,500 ± 100
280 ± 10
14,000 ± 4,000

232
700 ± 100
460 ± 60
42,000 ± 6,000
3,300 ± 600

233
3,200 ± 300
350 ± 80
>150,000
2,400 ± 700

234
3,000 ± 300
1,100 ± 400
24,000 ± 6,000
9,000 ± 1,000

235
3,500 ± 400
2,200 ± 400
49,000 ± 6,000
6,500 ± 1,600

236
49,000 ± 11,000
29,000 ± 5,000
48,000 ± 10,000

237
1,200 ± 300
730 ± 140
22,000 ± 1,000
6,600 ± 900

238
780 ± 190
57 ± 8
23,000 ± 2,000
4,700 ± 1,200

239
420 ± 60
70 ± 20
39,000 ± 3,000
28,000 ± 4,000

240
2,900 ± 100
1,300 ± 200
>24,000
1,300 ± 100

241
560 ± 90
110 ± 20
>28,000
18,000 ± 4,000

242
2,400 ± 400
580 ± 150
18,000 ± 2,000
2,900 ± 600

243
2,200 ± 500
670 ± 240
64,000 ± 10,000
26,000 ± 6,000

244
1,600 ± 400
150 ± 10
87,000 ± 11,000
35,000 ± 7,000

245
3,400 ± 1000
440 ± 90
79,000 ± 7,000
14,000 ± 1,700

246
2,800 ± 260
1,900 ± 450
14,000 ± 2,000
6,200 ± 1,300

247
6,100 ± 2,000
1,200 ± 250
10,000 ± 1,400
7,100 ± 1,700

248
830 ± 100
200 ± 25
23,000 ± 1,000
610 ± 120

249
4,100 ± 820
420 ± 100
18,000 ± 3,500
19,000 ± 3,800

250
99,000 ± 21,000
137,000 ± 14,000
>275,000
137,000 ± 10,000

251
128,000 ± 4,000
51,000 ± 1,000
>275,000
82,000 ± 8,000

252
>380,000
33,000 ± 3,000
>380,000
>380,000

253
>380,000
>38,000
>380,000
>380,000

254
2,700 ± 800
1,100 ± 100
43,000 ± 6,000
>65,000

255
2,900 ± 500
55 ± 2
119,000 ± 15,000
99,000 ± 4,000

256
1,500 ± 200
880 ± 200
7,500 ± 800
7,100 ± 300

257
2,800 ± 600
320 ± 30
25,000 ± 2,000
26,000 ± 3,000

258
>45,000
>45,000
>45,000
>45,000

259
16000 ± 3000
2400 ± 200
>85,000
4700 ± 400

260
1600 ± 500
150 ± 20
>64,000
19000 ± 4500

261
6300 ± 1100
1000 ± 150
64000 ± 2000
38000 ± 2100

262
8700 ± 1300
3900 ± 570
287000 ± 14000
73000 ± 17000

263
2000 ± 300
1400 ± 200
124000 ± 18000
39000 ± 7000

264
1400 ± 100
76 ± 17
>85,000
54000 ± 20000

265
810 ± 20
8 ± 1
1100 ± 200
250 ± 80

266
140 ± 20
70 ± 18
56000 ± 15000
32000 ± 7000

267
900 ± 160
160 ± 20
>90,000
28000 ± 8000

268
2100 ± 200
330 ± 90
54,000 ± 16000
>8,000

269
11000 ± 3000
850 ± 320
52000 ± 4000
>7,000

270
8000 ± 1500
1300 ± 100
>84,000
7100 ± 700

271
1700 ± 200
200 ± 90
>93,000
>9,300

272
>46,000
>4,700
>47,000
>4,700

273
30000 ± 5000
>1,500
>45,000
>4,500

274
39000 ± 3000
1200 ± 300
>46,000
>4,500

275
1500 ± 300
370 ± 40
>62,000
>6,200

276
1500 ± 200
760 ± 100
>61,000
>6,100

277
760 ± 70
190 ± 20
31,000 ± 6000
9,800 ± 1000

278
1000 ± 100
270 ± 10
>94000
>9,400

279
1700 ± 400
190 ± 20
>90000
>9,000

280
2400 ± 800
<80
>83000
>2,800

281
1800 ± 700
170 ± 10
27000 ± 2000
5000 ± 700

282
680 ± 60
110 ± 10
>85000
>8,500

283
2900 ± 1200
300 ± 20
40000 ± 4000
>9,300

284
13,600 (N = 2)
340 ± 20

>8,800

285
3800 ± 1100
310 ± 20
84000 ± 9000
2000 ± 100

286
48000 ± 10000
6300 ± 200
51000 ± 1700
>8,800

287
455000 ± 22000
1100 ± 100
567000 ± 17000
4700 ± 400

288
1800 ± 600
150 ± 20
11000 ± 3200
−9,000

289
51 ± 4
530 ± 150
>290000
>8,800

294
960 ± 170

295
200 ± 44

296
250 (N = 2)

297
2200 (N = 1)

298
>7000

TALE 14

The IUPAC name of Illudofulvene Analogs.

Illudofulvene

Entry #
Analog
IUPAC Name of Illudofulvene Analog

1
001
(R)-6′-hydroxy-2′,4′,6′-trimethylspiro[cyclopropane-1,5′-inden]-

7′(6′H)-one

2
002
(6′R)-6′-hydroxy-3′-(hydroxymethyl)-2′,4′,6′-trimethylspiro

[cyclopropane-1,5′-inden]-7′(6′H)-one

3
003
(6′R,6′′′R)-3′,3′′′-methylenebis(6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one)

4
004
(R)-3′-bromo-6′-hydroxy-2′,4′,6′-trimethylspiro[cyclopropane-1,5′-

inden]-7′(6′H)-one

5
005
(R)-6′-hydroxy-3′-iodo-2′,4′,6′-trimethylspiro[cyclopropane-1,5′-

inden]-7′(6′H)-one

6
006
(R)-6′-hydroxy-3′-(4-hydroxybenzyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

7
007
(R)-6′-hydroxy-3′-(4-methoxybenzyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

8
008
(R)-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden]-3′-yl)methoxy)methyl acetate

9
009
(R)-6′-hydroxy-3′-(3-hydroxypropyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

10
010
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propanal

11
011
(R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-indene]-3′-carbaldehyde

12
012
(R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-nitrospiro[cyclopropane-1,5′-

inden]-7′(6′H)-one

13
013
4-hydroxy-5-(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)cyclohexane-

1,3-dicarbaldehyde

14
014
(4a′S,7′R,9b′S)-7′-hydroxy-4a′,7′,9′-trimethyl-4a′,9b′-dihydro-4′H-

spiro[cyclopropane-1,8′-indeno[1,2-d][1,3]dioxin]-6′(7′H)-one

15
015
(R)-3′-(hydroxymethyl)-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-6′-yl acetate

16
016
(R)-3′-(ethoxymethyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

17
017
(6′R,6′′′R)-3′,3′′′-(oxybis(methylene))bis(6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one)

18
018
(R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-((((2R,3S,4R,5R,6S)-3,4,5-

trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-

yl)oxy)methyl)spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

19
019
(6′R)-3′-((2,3-dihydroxypropoxy)methyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

20
020
(R)-3′-((2-bromoethoxy)methyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

21
021
(R)-6′-hydroxy-3′-(((2-methoxypropan-2-yl)oxy)methyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

22
022
(R)-6′-hydroxy-3′-((2-hydroxyethoxy)methyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

23
023
(R)-6′-hydroxy-3′-(((4-hydroxyphenyl)thio)methyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

24
024
(R)-3′-((benzylthio)methyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

25
025
methyl (R)-2-(((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)thio)acetate

26
026
(R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-((p-tolylthio)methyl)

spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

27
027
(R)-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden]-3′-yl)methyl phenyl carbonate

28
028
(R)-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl benzoate

29
029
(R)-2-(((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)thio)acetic acid

30
030
methyl (R)-2-(((6′-hydroxy-1′4(2-methoxy-2-oxoethyl)thio)-

2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro[cyclopropane-1,5′-

inden]-3′-yl)methyl)thio)acetate

31
031
methyl 2-((((6′R)-6′,7a′-dihydroxy-1′4(2-methoxy-2-

oxoethyl)thio)-2′,4′,6′-trimethyl-7′-oxo-1′,6′,7′,7a′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)thio)acetate

32
032
(6′R)-3′-(((2,3-dihydroxypropyl)thio)methyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

33
033
7′-methyl-4′H-dispiro[cyclopropane-1,6′-indene-5′,2″-

[1,3]dioxolan1-4′-one

34
034
(R)-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl acetate

35
035
6′-hydroxy-4′-methylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

36
036
(R)-3′-((1H-imidazol-1-yl)methyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

37
037
1-carboxy-2-((((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)thio)ethan-1-

aminium

38
038
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propanoic acid

39
039
(R)-3′-(3,3-dimethoxypropyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

40
040
(R)-3′-(3,3-diethoxypropyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

41
041
(R,Z)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)acrylaldehyde

42
042
(R)-3′-(hydroxymethyl)-4′,6′-dimethyl-6′-((triethylsilyl)oxy)

spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

43
043
(R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-(((triethylsilyl)oxy)methyl)

spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

44
044
(R)-2′,4′,6′-trimethyl-6′-((triethylsilyl)oxy)-3′-(((triethylsilyl)

oxy)methyl)spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

45
045
methyl 2-((7-hydroxy-5-(2-hydroxyethyl)-3-(hydroxymethyl)-

2,4,6-trimethyl-1H-inden-1-yl)thio)acetate

46
046
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl acetate

47
047
(6′R)-3′-(2-(1,7-dihydroxy-2,4,6-trimethyl-1H-inden-5-yl)ethyl)-6′-

hydroxy-2′,4′,6′-trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

48
048
(R)-6′-hydroxy-2′,4′,6′-trimethyl-1′-(p-tolylthio)-3′-((p-tolylthio)

methyl)spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

49
049
(R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-(p-tolylthio)spiro[cyclopropane-

1,5′-inden]-7′(6′H)-one

50
050
(R)-6′-hydroxy-2′,4′,6′-trimethyl-1′,3′-bis(p-tolylthio)spiro

[cyclopropane-1,5′-inden]-7′(6′H)-one

51
051
(R)-2-(2-(((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)thio)acetoxy)

ethyl 2-mercaptoacetate

52
052
ethane-1,2-diyl bis(2-((((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-

6′,7′-dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)thio)acetate)

53
053
(R)-3′-((2-(2-bromoethoxy)ethoxy)methyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

54
054
(R)-6′-hydroxy-1′-(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)-3′-

(hydroxymethyl)-2′,4′,6′-trimethylspiro[cyclopropane-1,5′-inden]-

7′(6′H)-one

55
055
5-(2-hydroxyethyl)-1-((4-hydroxyphenyethio)-3-(((4-

hydroxyphenyl)thio)methyl)-2,4,6-trimethyl-1H-inden-7-ol

56
056
(R)-6′-hydroxy-3′((4-hydroxyphenyl)thio)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

57
057
(R)-6′-hydroxy-1′((4-hydroxyphenyl)thio)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

58
058
(R)-6′-hydroxy-1′,3′-bis((4-hydroxyphenyl)thio)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

59
059
(6′S,7′R)-4′-methyl-6′-((triethylsilyl)oxy)-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden]-7′-ol

60
060
(R)-7′-methyl-4′H-dispiro[cyclopropane-1,6′-indene-5′,2″-

[1,3]dioxolan1-4′-ol

61
061
(S)-4′-methyl-6′-((triethylsilyl)oxy)spiro[cyclopropane-1,5′-

inden]-7′(6′H)-one

62
062
(R)-6′-hydroxy-2′-(hydroxymethyl)-4′,6′-dimethylspiro

[cyclopropane-1,5′-inden]-7′(6′H)-one

63
063
(R)-6′-hydroxy-2′,3′-bis(hydroxymethyl)-4′,6′-

dimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

64
064
N-acetyl-S-(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)-L-cysteine

65
065
(R)-2-acetamido-3-((((R)-6′-hydroxy-4′,6′-dimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)thio)-N-((S)-1-

phenylethyl)propanamide

66
066
(S)-2-acetamido-3-((((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)thio)-N-((S)-1-

phenylethyl)propanamide

67
067
4-methyl-2,3-dihydro-5H-indeno[5,6-b]furan-5-one

68
068
5-hydroxy-6-(2-hydroxyethyl)-7-methyl-1H-inden-1-one

69
069
5-(2-hydroxyethoxy)-6-(2-hydroxyethyl)-7-methyl-1H-inden-1-one

70
070
(3a′R,4′R)-4′-hydroxy-7′-methyl-3a′,4′-dihydro-1′H-

dispiro[cyclopropane-1,6′-indene-5′,2″-[1,3]dioxolan]-1′-one

71
071
(3a′R,4′R)-7′-methyl-4′-((triethylsilyl)oxy)-3a′,4′-dihydro-1′H-

dispiro[cyclopropane-1,6′-indene-5′,2″-[1,3]dioxolan]-1′-one

72
072
(7′R,7a′R)-7′-hydroxy-4′-methyl-7′,7a′-dihydrospiro[cyclopropane-

1,5′-indene]-3′,6′-dione

73
073
(7′R,7a′R)-4′-methyl-7′-((triethylsilyl)oxy)-7′,7a′-dihydrospiro

[cyclopropane-1,5′-indene]-3,6′-dione

74
074
(6′R)-3′-((((2,2-dimethyl-1,3-dioxolan-4-yl)methypthio)methyl)-6′-

hydroxy-2′,4′,6′-trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

75
075
(R)-(6′-hydroxy-4′,6′-dimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl acetate

76
076
(R)-(6′-hydroxy-4′,6′-dimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-indenel-2′,3′-diyl)bis(methylene)diacetate

77
077
(R)-(6′-hydroxy-3′-(hydroxymethyl)-4′,6′-dimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl acetate

78
078
(R)-(6′-hydroxy-2′-(hydroxymethyl)-4′,6′-dimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl acetate

79
079
(R)-6′-hydroxy-2′-(methoxymethyl)-4′,6′-dimethylspiro

[cyclopropane-1,5′-inden]-7′(6′H)-one

80
080
(R)-6′-hydroxy-3′-(methoxymethyl)-2′,4′,6′-trimethylspiro

[cyclopropane-1,5′-inden]-7′(6′H)-one

81
081
(R)-6′-hydroxy-2′-(hydroxymethyl)-3′-(methoxymethyl)-4′,6′-

dimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

82
082
(R)-6′-hydroxy-2′,3′-bis(methoxymethyl)-4′,6′-

dimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

83
083
(R)-2-acetamido-3-((((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)thio)-N-((S)-1-

((2-(((S)-4-methyl-1-oxopentan-2-yl)amino)-2-oxoethyl)amino)-1-

oxo-3-phenylpropan-2-yl)propanamide

84
084
(S)-2-((R)-2-acetamido-3-((((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-

oxo-6′,7′-dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)

thio)propanamido)-4-methyl-N-(2-oxo-2-(((R)-1-oxo-3-

phenylpropan-2-yl)amino)ethyl)pentanamide

85
085
(S)-2-(((R)-2-acetamido-3-4((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-

oxo-6′,7′-dihydrospiro[cyclopropane-1,5′-inden]-3′-

yl)methyl)thio)propanamido)-4-methyl-N-((S)-4-methyl-1-oxo-1-

(((R)-1-oxo-3-phenylpropan-2-yl)amino)pentan-2-yl)pentanamide

86
086
(R)-2-acetamido-3-((((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden-3-yl)methyl)thio)-N-(2-

oxo-2-(((R)-1-oxo-3-phenylpropan-2-yl)amino)ethyl)propanamide

87
087
(S)-2-((R)-2-acetamido-3-((((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-

oxo-6′,7′-dihydrospiro[cyclopropane-1,5′-inden-3′-yl)methyl)

thio)propanamido)-4-methyl-N-((R)-4-methyl-1-(((S)-4-

methyl-1-oxopentan-2-yl)amino)-1-oxopentan-2-yl)pentanamide

88
088
(R)-(6′-acetoxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden-3′-yl)methyl acetate

89
089
N5-((R)-1-((carboxymethyl)amino)-3-((((R)-6′-hydroxy-2′,4′,6′-

trimethyl-7′-oxo-6′,7′-dihydrospiro[cyclopropane-1,5′-inden-3′-

yl)methyl)thio)-1-oxopropan-2-yl)-D-glutamine

90
090
(R)-2′-(hydroxymethyl)-4′,6′-dimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-6′-yl acetate

91
091
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden-3′-yl)propanoic acid

92
092
(R)-6′-hydroxy-3′-(3-methoxypropyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

93
093
(R)-3′-(3,3-diethoxypropyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

94
094
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden-3′-yl)propyl acetate

95
095
(R)-6′-hydroxy-3′-(3-methoxypropyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

96
096
(R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-(((1-methyl-1H-imidazol-2-

yl)thio)methyl)spiro[cyclopropane-1,5′-inden-7′(6′H)-one

97
097
S-(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden-3′-yl)methyl)homocysteine

98
098
((S)-3-((((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden-3′-yl)methyl)thio)-2-

methylpropanoyl)proline

99
099
(2′S,6′R)-6′-hydroxy-2′-(hydroxymethyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-indene-3′,7′(2′H,6′H)-dione

100
100
S-(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden-3′-yl)methyl)-N-S-(((R)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro[cyclopropane-1,5′-

inden-3′-yl)methyl)cysteinyl-L-asparaginylglycyl-L-arginylcysteine

101
101
S-(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden-3′-yl)methyl)-N-S-(((R)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro[cyclopropane-1,5′-inden-

3′-yl)methyl)cysteinyl-L-arginylglycyl-L-asparaginylcysteine

102
102
S-(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden-3′-yl)methyl)cysteinyl-L-

asparaginylglycyl-L-arginylcysteine

103
103
(R)-(6′-acetoxy-2′-(hydroxymethyl)-4′,6′-dimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden-3′-yl)methyl acetate

104
104
(R)-8′-hydroxy-6′,8′-dimethyl-1,5′-dihydrospiro[cyclopropane-1,7′-

indeno[1,2-][1,3]dioxepin]-9′(8′H)-one

105
105
(E)-2-((2R,45)-4-hydroxy-2-((1R,25)-2-hydroxy-4,4-

dimethylcyclopentyl)-2-methylcyclobutylidene)propanal

106
106
5-(((3′S,6′R)-6′-hydroxy-2′,2′,4′,6′-tetramethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl)oxy)-5-

oxopentanoic acid

107
107
(3′S,6′R)-6′-hydroxy-2′,2′,4′,6′-tetramethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl ((13S)-13-methyl-

17-oxo-7,8,9,11,12,13,14,15,16,17-decahydro-6H-

cyclopenta[a]phenanthren-3-yl)glutarate

108
108
(13S)-17-hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-

decahydro-6H-cyclopenta[a]phenanthren-3-yl 3-((R)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro[cyclopropane-1,5′-

inden]-3′-yl)propanoate

109
109
(13S)-17-hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-

decahydro-6H-cyclopenta[a]phenanthren-3-yl ((3′S,6′R)-6′-

hydroxy-2′,2′,4′,6′-tetramethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl)glutarate

110
110
(13S)-13-methyl-17-oxo-7,8,9,11,12,13,14,15,16,17-decahydro-

6H-cyclopenta[a]phenanthren-3-yl 3-((R)-6′-hydroxy-2′,4′,6′-

trimethyl-7′-oxo-6′,7′-dihydrospiro[cyclopropane-1,5′-inden]-3′-

yl)propanoate

111
111
(10R,13S)-10,13-dimethyl-3-oxo-

2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-

cyclopenta[a]phenanthren-17-yl 3-(6′-hydroxy-2′,4′-dimethyl-7′-

oxo-6′,7′-dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propanoate

112
112
(13S)-10,13-dimethyl-17-oxohexadecahydro-1H-

cyclopenta[a]phenanthren-3-yl 3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-

7′-oxo-6′,7′-dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propanoate

113
113
(R)-3′-(but-3-en-1-yl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

114
114
(6′R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-(2-(oxiran-2-

yl)ethyl)spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

115
115
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propanal oxime

116
116
(R)-3′-(tert-butoxymethyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

117
117
5-(((2′S,6′R)-3′-((4-carboxybutanoyl)oxy)-6′-hydroxy-2′,4′,6′-

trimethyl-7′-oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-

inden]-2′-yl)methoxy)-5-oxopentanoic acid

118
118
5-(((2′S,6′R)-2′-(((3,5-dinitrobenzoyl)oxy)methyl)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-

1,5′-inden]-3′-yl)oxy)-5-oxopentanoic acid

119
119
(6′R)-3′-(3,4-dihydroxybutyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

120
120
(R)-6′-hydroxy-3′-(3-((34(S)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-

6′,7′-dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propylidene)

hydrazineylidene)propyl)-2′,4′,6′-trimethylspiro[cyclopropane-

1,5′-inden]-7′(6′H)-one

121
121
(R)-2-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden]-3′-yl)propylidene)hydrazine-1-

carboxamide

122
122
(R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-(3-(2-phenylhydrazineylidene)

propyl)spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

123
123
(R)-N′-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propylidene)-4-

methylbenzenesulfonohydrazide

124
124
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propanal O-acetyl

oxime

125
125
(R)-3′-(3-(2-(2,4-dinitrophenyl)hydrazineylidene)propyl)-6′-

hydroxy-2′,4′,6′-trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-

one

126
126
(R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-indenel-3′-carbaldehyde oxime

127
127
2-hydroxy-4(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)butanenitrile

128
128
(6′R)-6′-hydroxy-3′-(3-hydroxybutyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

129
129
(6′R)-2′,4′,6′-trimethyl-6′,7′-dihydrospiro[cyclopropane-1,5′-

indenel-6′,7′-diol

130
130
(R)-6′-hydroxy-3′-(3-(hydroxyamino)propyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

131
131
(R)-N-benzyl-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propanamide

132
133
(E)-7-(chloromethylene)-5-hydroxy-5,9-dimethylspiro[3.5]non-8-

en-6-one

133
134
(E)-6-(chloromethylene)-4-hydroxy-4,8-dimethylspiro[2.5]oct-7-

en-5-one

134
135
((2′S,6′R)-3′,6′-dihydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl 4-

nitrobenzoate

135
136
((2′S,6′R)-3′,6′-dihydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl 4-(N-

acetoxyacetamido)benzoate

136
137
((2′S,6′R)-6′-hydroxy-2′,4′,6′-trimethyl-3′,7′-dioxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl 4-

nitrobenzoate

137
138
((2′S,6′R)-6′-hydroxy-2′,4′,6′-trimethyl-3′,7′-dioxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl 4-(N-

acetoxyacetamido)benzoate

138
139
(2′S,6′R)-6′-hydroxy-2′,4′,6′-trimethyl-2′-(((4-

nitrobenzoyl)oxy)methyl)-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl 4-nitrobenzoate

139
140
((2′S,6′R)-3′-((4-(N-acetoxyacetamido)benzoyl)oxy)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-

1,5′-inden]-2′-yl)methyl 4-(N-acetoxyacetamido)benzoate

140
141
dimethyl (5′R)-4′,5′-dihydroxy-5′,7′,9′-trimethyl-4′,5′-dihydro-1′H-

spiro[cyclopropane-1,6′-[1,3a]ethenoindenel-2′,3′-dicarboxylate

141
142
dimethyl (5′R)-5′-hydroxy-5′,7′,9′-trimethyl-4′-oxo-4′,5′-dihydro-

1′H-spiro[cyclopropane-1,6′-[1,3a]ethenoindenel-2′,3′-dicarboxylate

142
143
(R)-6′-hydroxy-1′,2′,4′,6′-tetramethylspiro[cyclopropane-1,5′-

inden]-7′(6′H)-one

143
144
(R)-2-((2′-ethyl-6′-hydroxy-4′,6′-dimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methoxy)ethyl acetate

144
145
(R)-54(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methoxy)-5-

oxopentanoic acid

145
146
(R)-4-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)butanenitrile

146
147
(R)-3′-((benzo[d]thiazol-2-ylthio)methyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

147
148
(R)-3′-((benzo[d]oxazol-2-ylthio)methyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

148
149
(R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-(((1-methyl-1H-tetrazol-5-

yl)thio)methyl)spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

149
150
(R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-(((5-methyl-1H-benzo[d]

imidazol-2-yl)thio)methyl)spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

150
151
(R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-(((1-phenyl-1H-tetrazol-5-

yl)thio)methyl)spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

151
152
(R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-(((5-nitro-1H-benzo[d]imidazol-

2-yl)thio)methyl)spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

152
153
(R)-3′-(((1H-1,2,4-triazol-3-yl)thio)methyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

153
154
(R)-6′-hydroxy-3′-(((4-hydroxypteridin-2-yl)thio)methyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

154
155
(R)-6′-hydroxy-3′-(((1-(4-hydroxyphenyl)-1H-tetrazol-5-

yl)thio)methyl)-2′,4′,6′-trimethylspiro[cyclopropane-1,5′-inden]-

7′(6′H)-one

155
156
(R)-4-(5-(((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)thio)-1H-

tetrazol-1-yl)phenyl acetate

156
157
7′-methyl-4′H-dispiro[cyclobutane-1,6′-indene-5′,2″-[1,3]dioxolan]-

4′-one

157
158
5-hydroxy-2,2,6,8a-tetramethyl-2,3,3a,8,8a,8b-hexahydro-1H-

cyclobuta[d]cyclopenta[b]oxepin-7(5H)-one

158
159
((6′R)-3′,6′-dihydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl acetate

159
160
5-(((6′R)-2′-(acetoxymethyl)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-

2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl)oxy)-5-

oxopentanoic acid

160
161
((6′R)-6′-hydroxy-2′,4′,6′-trimethyl-3′,7′-dioxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl acetate

161
162
5-(((6′R)-6′-hydroxy-2′,4′,6′-trimethyl-3′,7′-dioxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methoxy)-5-

oxopentanoic acid

162
163
(6′R)-2′-(acetoxymethyl)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-

2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl 2-

chloroacetate

163
164
(R)-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl 2-chloroacetate

164
165
(R)-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden]-3′-yl)methyl 2-morpholinoacetate

165
166
(6′R)-2′-(acetoxymethyl)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-

2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl methyl

glutarate

166
167
(6′R)-6′-hydroxy-2′-(hydroxymethyl)-2′,4′,6′-trimethyl-7′-oxo-

2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl methyl

glutarate

167
168
((6′R)-6′-hydroxy-2′,4′,6′-trimethyl-3′,7′-dioxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl methyl

glutarate

168
169
((6′R)-6′-hydroxy-2′,4′,6′-trimethyl-3′,7′-dioxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl 2-

chloroacetate

169
170
5-(2-hydroxyethyl)-3-(hydroxymethyl)-2,4,6-trimethyl-1H-inden-

7-ol

170
171
6-(2-hydroxyethyl)-2,5,7-trimethyl-1-methylene-1H-inden-4-ol

171
172
6-ethyl-2,5,7-trimethyl-1-methylene-1H-inden-4-ol

172
173
2-(4-hydroxy-2,5,7-trimethyl-1-methylene-1H-inden-6-yl)ethyl

acetate

173
174
5-(2-hydroxyethyl)-3-(hydroxymethyl)-2,4,6-trimethyl-1H-indene-

1,7-diol

174
175
(2S,3S,4R,5S,6R)-2-(acetoxymethyl)-6-(((R)-6′-hydroxy-2′,4′,6′-

trimethyl-7′-oxo-6′,7′-dihydrospiro[cyclopropane-1,5′-inden]-3′-

yl)methoxy)tetrahydro-2H-pyran-3,4,5-triyltriacetate

175
176
3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl leucinate

176
177
(R)-5-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propoxy)-5-

oxopentanoic acid

177
178
(R)-4-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propoxy)-4-

oxobutanoic acid

178
179
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl glycinate

179
180
(1a′R,3′R,7′S,7a′R)-3′,7′-dihydroxy-1a′,3′,6′,6′-tetramethyl-6′,7′-

dihydro-1a′H-spiro[cyclopropane-1,2′-indeno[3a,4-b]oxiren]-

4′(3′H)-one

180
181
((1a′R,3′R,6′S,7′S,7a′R)-3′,7′-dihydroxy-1a′,3′,6′-trimethyl-4′-oxo-

3′,4′,6′,7′-tetrahydro-1a′H-spiro[cyclopropane-1,2′-indeno[3a,4-

b]oxiren]-6′-yl)methyl acetate

181
182
(2′R,7′S,7a′S)-2′-chloro-7′-hydroxy-2′,4′-dimethyl-1′,2′,7′,7a′-

tetrahydrospiro[cyclopropane-1,5′-indene]-3′,6′-dione

182
183
(2′S,7′S,7a′S)-7′-hydroxy-2′-isopropoxy-2′,4′-dimethyl-1′,2′,7′,7a′-

tetrahydrospiro[cyclopropane-1,5′-indene]-3′,6′-dione

183
184
(R)-1-hydroxy-1-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)urea

184
185
(S)-6′-hydroxy-2′,4′,6′-trimethylspiro[cyclopropane-1,5′-inden]-

7′(6′H)-one

185
186
(6′S,7′R)-6′,7′-dihydroxy-2′,4′,6′-trimethyl-7′,7a′-

dihydrospiro[cyclopropane-1,5′-inden]-3′(6′H)-one

186
187
(S)-6′-hydroxy-3′-(hydroxymethyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

187
188
(6′S,6S)-3′,3-methylenebis(6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one)

188
189
(R)-14(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)-1H-pyrrole-

2,5-dione

189
190
(R)-1-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl)-1H-pyrrole-

2,5-dione

190
191
6′-hydroxy-4′,6′-dimethylspiro[cyclobutane-1,5′-inden]-7′(6′H)-one

191
192
(R)-2-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)isoindoline-

1,3-dione

192
193
(R)-3′-(azidomethyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

193
194
(R)-3′-(((R)-3′-(azidomethyl)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-

6′,7′-dihydrospiro[cyclopropane-1,5′-inden]-1′-yl)methyl)-6′-

hydroxy-2′,4′,6′-trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-

one

194
195
(R)-3′-(3-azidopropyl)-6′-hydroxy-4′,6′-

dimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

195
196
3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl L-prolinate

196
197
(R)-2-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden]-3′-yl)propyl)isoindoline-1,3-dione

197
198
(R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-((4-

nitrophenoxy)methyl)spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

198
199
(R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-

(phenoxymethyl)spiro[cyclopropane-1,5′-inden]-7′(6′H)-one

199
200
(R)-6′-hydroxy-3′-(2-hydroxybenzyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

200
201
(R)-N-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl)acetamide

201
202
(S)-N-(3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden]-3′-yl)propyl)pyrrolidine-2-carboxamide

202
203
3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl L-seryl-L-

prolinate

203
204
2′-(((tert-butyldimethylsilyl)oxy)methyl)-3′,6′-dihydroxy-2′,4′,6′-

trimethyl-2′,3′-dihydrospiro[cyclopropane-1,5′-inden]-7′(6′H)-one

204
205
(2′R,3′S,6′R)-3′-amino-2′-(((tert-butyldimethylsilyl)oxy)methyl)-6′-

hydroxy-2′,4′,6′-trimethyl-2′,3′-dihydrospiro[cyclopropane-1,5′-

inden]-7′(6′H)-one

205
206
(2′R,3′S,6′R)-3′-amino-6′-hydroxy-2′-(hydroxymethyl)-2′,4′,6′-

trimethyl-2′,3′-dihydrospiro[cyclopropane-1,5′-inden]-7′(6′H)-one

206
207
(S)-2-amino-N-(3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl)-4-

methylpentanamide

207
208
3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden]-3′-yl)propyl (tert-butoxycarbonyl)-

L-seryl-L-prolinate

208
209
3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl L-seryl-L-

seryl-L-prolinate

209
210
3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl ((S)-2-((S)-2-

((S)-2-((2S,4R)-1-acetyl-4-hydroxypyrrolidine-2-carboxamido)-3-

hydroxypropanamido)-3-hydroxypropanamido)-2-

cyclohexylacetyl)-L-glutaminyl-L-seryl-L-seryl-L-prolinate

210
211
(R)-3′-(3-aminopropyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

211
212
(3′S,6′R)-6′-hydroxy-2′,2′,4′,6′-tetramethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl L-prolinate

212
213
(2′S,3′R,6′R)-2′-(((tert-butyldimethylsilyl)oxy)methyl)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-

1,5′-inden]-3′-yl L-prolinate

213
214
(2′S,3′R,6′R)-2′-(((tert-butyldimethylsilyl)oxy)methyl)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-

1,5′-inden]-3′-yl L-seryl-L-seryl-L-prolinate

214
215
(2′S,3′R,6′R)-2′-(((tert-butyldimethylsilyl)oxy)methyl)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-

1,5′-inden]-3′-yl ((S)-2-((S)-2-((S)-2-((25,4R)-1-acetyl-4-

hydroxypyrrolidine-2-carboxamido)-3-hydroxypropanamido)-3-

hydroxypropanamido)-2-cyclohexylacetyl)-L-glutaminyl-L-seryl-

L-seryl-L-prolinate

215
216
(3′S,6′R)-6′-hydroxy-2′,2′,4′,6′-tetramethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl ((S)-2-((S)-2-((S)-2-

((2S,4R)-1-acetyl-4-hydroxypyrrolidine-2-carboxamido)-3-

hydroxypropanamido)-3-hydroxypropanamido)-2-

cyclohexylacetyl)-L-glutaminyl-L-seryl-L-seryl-L-prolinate

216
217
(R)-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl 6-(2,5-dioxo-

2,5-dihydro-1H-pyrrol-1-yl)hexanoate

217
218
(3′S,6′R)-6′-hydroxy-2′,2′,4′,6′-tetramethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl 6-(2,5-dioxo-2,5-

dihydro-1H-pyrrol-1-yl)hexanoate

218
219
(2′S,3′R,6′R)-2′-(((tert-butyldimethylsilyl)oxy)methyl)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-

1,5′-inden]-3′-yl 4-(fluorosulfonyl)benzoate

219
220
(R)-1-acetoxy-1-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)urea

220
221
(S)-2-((3S,6S,9S,12S,15S)-3-((1H-imidazol-4-yl)methyl)-12-(4-

aminobutyl)-6,9-bis(hydroxymethyl)-15-isobutyl-1-morpholino-

1,4,7,10,13-pentaoxo-2,5,8,11,14-pentaazahexadecan-16-amido)-

N1-((S)-1-((3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl)amino)-4-

methyl-1-oxopentan-2-yl)pentanediamide

221
222
(3′S,6′R)-6′-hydroxy-2′,2′,4′,6′-tetramethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl 4-

(fluorosulfonyl)benzoate

222
223
3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl ((4R,7S,13S)-

13-(2-amino-2-oxoethyl)-7-(3-guanidinopropyl)-6,9,12,15-

tetraoxo-1,2-dithia-5,8,14-triazacycloheptadecane-4-

carbonyl)glycinate

223
224
(R,E)-6′-hydroxy-2′,4′,6′-trimethylspiro[cyclopropane-1,5′-inden]-

7′(6′H)-one oxime

224
225
(2′R,3′R,6′R,E)-2′,3′,6′-trihydroxy-2′,4′,6′-trimethyl-2′,3′-

dihydrospiro[cyclopropane-1,5′-inden]-7′(6′H)-one oxime

225
226
(R)-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden]-3′-yl)methyl (E)-octadec-9-enoate

226
227
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden]-3′-yl)propyl (E)-octadec-9-enoate

227
228
(2′S,6′R)-6′-hydroxy-2′,4′,6′-trimethyl-2′-((((E)-octadec-9-

enoyl)oxy)methyl)-7′-oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-

1,5′-inden]-3′-yl (E)-octadec-9-enoate

228
229
(R,E)-N-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl)octadec-9-

enamide

229
230
N-((3′R,6′R)-6′-hydroxy-2′,2′,4′,6′-tetramethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl)methanesulfonamide

230
231
N-((3′R,6′R)-6′-hydroxy-2′,2′,4′,6′-tetramethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl)-4-

methylbenzenesulfonamide

231
232
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl

hydroxycarbamate

232
233
ethyl (R)-hydroxy((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)carbamate

233
234
benzyl (R)-hydroxy((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)carbamate

234
235
tert-butyl (R)-hydroxy((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)carbamate

235
236
(R)-6′-hydroxy-3′-(hydroxymethyl)-1′,2′,4′,6′-

tetramethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

236
237
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl (2-

bromoethyl)carbamate

237
238
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl (2-

chloroethyl)carbamate

238
239
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl (2-

hydroxyethyl)carbamate

239
240
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl

acetoxy(acetyl)carbamate

240
241
(R)-N-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl)

methanesulfonamide

241
242
(R)-N-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl)-4-

methylbenzenesulfonamide

242
243
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl (2-

fluoroethyl)carbamate

243
244
(R)-1-hydroxy-1-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl)urea

245
(R)-1-hydroxy-1-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

244

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl)thiourea

246
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

245

[cyclopropane-1,5′-inden]-3′-yl)propyl morpholine-4-carboxylate

246
247
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden]-3′-yl)propyl morpholine-4-carboxylate

247
248
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl [1,4′-

bipiperidine]-1′-carboxylate

248
249
(R)-6′-hydroxy-2′-(hydroxymethyl)-3′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

249
250
((1a′R,2′S,3′R,6′R,7a′S)-3′-acetoxy-6′-hydroxy-2′,4′,6′-trimethyl-7′-

oxo-1′,1a′,2′,3′,6′,7′-hexahydrospiro[cyclopropane-1,5′-

cyclopropa[c]inden]-2′-yl)methyl acetate

250
251
((1a′S,2′S,3′R,6′R,7a′R)-3′-acetoxy-6′-hydroxy-2′,4′,6′-trimethyl-7′-

oxo-1′,1a′,2′,3′,6′,7′-hexahydrospiro[cyclopropane-1,5′-

cyclopropa[c]inden]-2′-yl)methyl acetate

251
252
(1a′R,3′S,6′R,7a′S)-3′,6′-dihydroxy-2′,2′,4′,6′-tetramethyl-1′,1a′,2′,3′-

tetrahydrospiro[cyclopropane-1,5′-cyclopropa[c]inden]-7′(6′H)-one

252
253
(1a′S,3′S,6′R,7a′R)-3′,6′-dihydroxy-2′,2′,4′,6′-tetramethyl-1′,1a′,2′,3′-

tetrahydrospiro[cyclopropane-1,5′-cyclopropa[c]inden]-7′(6′H)-one

253
254
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl 4-

sulfamoylbenzoate

254
255
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl sulfamate

255
256
3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl 1-(3-((R)-6′-

hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro[cyclopropane-

1,5′-inden]-3′-yl)propyl)-1H-1,2,3-triazole-4-carboxylate

256
257
3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl 1-(((R)-6′-

hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro[cyclopropane-

1,5′-inden]-3′-yl)methyl)-1H-1,2,3-triazole-4-carboxylate

257
258
(4-carboxy-4-(4-carboxy-4-((3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-

oxo-6′,7′-dihydrospiro[cyclopropane-1,5′-inden]-3′-

yl)propyl)amino)butanamido)butanoyl)glutamic acid

258
259
(R)-3′-((S)-2,2-dioxido-1,2,3-oxathiazinan-4-yl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

259
260
(R)-N-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl)-[1,4′-

bipiperidine]-l′-carboxamide

260
261
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl 1H-imidazole-

1-carboxylate

261
262
methyl (R)-2-4(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)sulfonyl)acetate

262
263
methyl 2-((((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)sulfinyl)acetate

263
264
N-[3-(6′-hydroxy-2′,6′-dimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl]sulfuric

diamide

264
265
N-hydroxy-N-l3-(6′-hydroxy-2′,6′-dimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl]sulfuric

diamide

265
266
N-[3-(6′-hydroxy-2′,6′-dimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl]-N-

methoxysulfuric diamide

266
267
(R)-2-amino-N-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)-N-

methoxyacetamide

267
268
(R)-2,2,2-trifluoro-N-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl)acetamide

268
269
(R)-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl (4-

methoxyphenyl)sulfamate

269
270
(R)-3′-(aminomethyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

270
271
(R)-N-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-dihydrospiro

[cyclopropane-1,5′-inden]-3′-yl)methyl)methanesulfonamide

271
272
(5S,6S,7S)-3-(((3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propanoyl)oxy)

methyl)-8-oxo-7-(2-(thiophen-2-yl)acetamido)-5-thia-1-

azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid 5-oxide

272
273
(5S,6S,7S)-3-((((3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-

yl)propyl)carbamoyl)oxy)methyl)-8-oxo-7-(2-(thiophen-2-

yl)acetamido)-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic

acid 5-oxide

273
274
(6S,7S)-3-((((3-((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-

yl)propyl)carbamoyl)oxy)methyl)-8-oxo-7-(2-(thiophen-2-

yl)acetamido)-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic

acid 5,5-dioxide

274
275
N-(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)pyrrolidine-2-

carboxamide

275
276
2-amino-N-(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)-4-

methylpentanamide

276
277
(R)-1-hydroxy-3-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)urea

277
278
(R)-14(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)-3-

methoxyurea

278
279
(R)-14(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3-yl)methyl)-3-(2-

hydroxyethyl)urea

279
280
(R)-1-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3-yl)propyl)-3-(2-

hydroxyethyl)urea

280
281
(R)-1-(2-chloroethyl)-3-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-

6,7′-dihydrospiro[cyclopropane-1,5′-inden]-3-yl)propyl)urea

281
282
(R)-1-(2-chloroethyl)-3-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)urea

282
283
N-1(6′-hydroxy-2′,6′-dimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl]sulfuric

diamide

283
284
(R)-3-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3-yl)methyl)pyrimidine-

2,4(1H,3H)-dione

284
285
N-hydroxy-N-1(6′-hydroxy-2′,6′-dimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl]sulfuric

diamide

285
286
(R)-5-fluoro-1-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)pyrimidine-

2,4(1H,3H)-dione

286
287
(R)-5-fluoro-3-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)pyrimidine-

2,4(1H,3H)-dione

287
288
(R)-1-hydroxy-3-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3-yl)propyl)urea

288
289
(R)-1-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)pyrimidine-

2,4(1H,3H)-dione

289
290
((S)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl 5-oxo-5-(((S)-

1,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydrobenzo[a]heptalen-7-

yl)amino)pentanoate

290
291
3-((S)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3-yl)propyl 5-oxo-5-(((S)-

1,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydrobenzo[a]heptalen-7-

yl)amino)pentanoate

291
292
N1-(((S)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3-yl)methyl)-N5-((S)-

1,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydrobenzo[a]heptalen-7-

yl)glutaramide

292
293
3-((S)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3-yl)-N-((S)-1,2,3,10-

tetramethoxy-9-oxo-5,6,7,9-tetrahydrobenzo[a]heptalen-7-

yl)propanamide

293
294
2-amino-N-(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)-N-

methoxypropanamide

294
295
2-amino-N-(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)-N-methoxy-4-

methylpentanamide

295
296
2-amino-N-(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)-N-methoxy-4-

(methylthio)butanamide

296
297
2-amino-N-(((R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)-3-(1H-indol-

3-yl)-N-methoxypropanamide

297
298
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl (tert-

butoxycarbonyl)glycinate

298
299
(2′S,3′R,6′R)-2′-(azidomethyl)-3′,6′-dihydroxy-2′,4′,6′-trimethyl-

2′,3′-dihydrospiro[cyclopropane-1,5′-inden]-7′(6′H)-one

299
300
(2′R,3′R,6′R)-3′-azido-6′-hydroxy-2′-(hydroxymethyl)-2′,4′,6′-

trimethyl-2′,3′-dihydrospiro[cyclopropane-1,5′-inden]-7′(6′H)-one

300
301
(R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl 6-oxo-6-

phenylhexanoate

301
302
((2′S,3′R,6′R)-3′,6′-dihydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl 3,5-

dinitrobenzoate

302
303
(2′S,3′R,6′R)-2′-(((3,5-dinitrocyclohexa-2,4-diene-1-

carbonyl)oxy)methyl)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-

2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl 3,5-

dinitrobenzoate

303
304
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl propiolate

304
305
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl (4-nitrophenyl)

carbonate

305
306
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl 4-

methylbenzenesulfonate

306
307
(3′R,6′R)-3′-azido-6′-hydroxy-2′,2′,4′,6′-tetramethyl-2′,3′-

dihydrospiro[cyclopropane-1,5′-inden]-7′(6′H)-one

307
308
(3′R,6′R)-3′-amino-6′-hydroxy-2′,2′,4′,6′-tetramethyl-2′,3′-

dihydrospiro[cyclopropane-1,5′-inden]-7′(6′H)-one

308
309
(2′R,3′S,6′R)-3′-azido-2′-(((tert-butyldimethylsilyl)oxy)methyl)-6′-

hydroxy-2′,4′,6′-trimethyl-2′,3′-dihydrospiro[cyclopropane-1,5′-

inden]-7′(6′H)-one

309
310
(R)-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl 4-

sulfamoylbenzoate

310
311
(3′S,6′R)-6′-hydroxy-2′,2′,4′,6′-tetramethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl 4-

sulfamoylbenzoate

311
312
(R)-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl 4-

methylbenzenesulfonate

312
313
2,3,4,5,6-pentafluoro-N-((3′R,6′R)-6′-hydroxy-2′,2′,4′,6′-

tetramethyl-7′-oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-

inden]-3′-yl)benzenesulfonamide

313
314
(R)-3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl 4-

methylbenzenesulfonate

314
315
(R)-3′-((2,5-dimethyl-1H-pyrrol-3-yl)methyl)-6′-hydroxy-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

315
316
((2′S,3′R,6′R)-3′,6′-dihydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl 4-

(fluorosulfonyl)benzoate

316
317
(2′S,3′R,6′R)-6′-hydroxy-2′-(hydroxymethyl)-2′,4′,6′-trimethyl-7′-

oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl 4-

(fluorosulfonyl)benzoate

317
318
((2′S,3′R,6′R)-3′-((4-(fluorosulfonyl)benzoyl)oxy)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-

1,5′-inden]-2′-yl)methyl 4-(fluorosulfonyebenzoate

318
332
(R)-6′-hydroxy-3′-((methoxyamino)methyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

319
333
((2′S,3′R,6′R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-(((4-

nitrophenoxy)carbonyl)oxy)-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl acetate

320
334
((2′S,3′R,6′R)-3′-(((2-((tert-

butoxycarbonyl)(methyl)amino)ethyl)(methyl)carbamoyl)oxy)-6′-

hydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl acetate

321
335
tert-butyl ((2′S,3′R,6′R)-6′-hydroxy-2′-(hydroxymethyl)-2′,4′,6′-

trimethyl-7′-oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-

inden]-3′-yl)ethane-1,2-diylbis(methylcarbamate)

322
337
4-nitrophenyl (R)-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)carbamate

323
338
4-nitrophenyl (R)-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl)carbamate

324
339
((2′S,3′R,6′R)-3′,6′-dihydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl 4-

methylbenzenesulfonate

325
340
(R)-N-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)

methanesulfonamide

326
345
(2′S,3′R,6′R)-2′-(acetoxymethyl)-6′-hydroxy-2′,4′,6′-trimethyl-7′-

oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl 4-(N-

(2-(2-(2-(2-aminoacetamido)acetamido)acetamido)ethyl)-N-

methylsulfamoyl)benzoate

327
346
((2′S,3′R,6′R)-3′,6′-dihydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl 4-(N-(2-(2-

(2-(2-aminoacetamido)acetamido)acetamido)ethyl)-N-

methylsulfamoyl)benzoate

328
347
((2′S,3′R,6′R)-3′,6′-dihydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl 4-

sulfamoylbenzoate

330
348
(3′S,6′R)-6′-hydroxy-2′,2′,4′,6′-tetramethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl4-

sulfamoylbenzoate

331
351
(3′S,6′R)-6′-hydroxy-2′,2′,4′,6′-tetramethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl4-(N-(2-(2-(2-(2-

aminoacetamido)acetamido)acetamido)ethyl)-N-

methylsulfamoyl)benzoate

332
353
((6′R)-3′-(((2-((((4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-

pyrrol-1-yl)hexanamido)-3-methylbutanamido)-5-

ureidopentanamido)benzyl)oxy)carbonyl)(methyl)amino)ethyl)

(methyl)carbamoyl)oxy)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-

2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl

acetate

333
354
4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-

yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)

benzyl (2-(34(S)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propylidene)

hydrazine-1-carbonyl)carbamate

334
356
((2′S,3′S,6′R)-3′-(((1-(9H-fluoren-9-yl)-13-methyl-3,6,9,12-

tetraoxo-2-oxa-4,7,10,13-tetraazapentadecan-15-

yl)(methyl)carbamoyl)oxy)-6′-hydroxy-2′,4′,6′-trimethyl-7′-

oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-2′-

yl)methyl acetate

335
357
((2′S,3′S,6′R)-3′-(((2-(2-(2-(2-aminoacetamido)acetamido)-N-

methylacetamido)ethyl)(methyl)carbamoyl)oxy)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-tetrahydrospiro

[cyclopropane-1,5′-inden]-2′-yl)methyl acetate

336
359
((2′S,3′S,6′R)-6′-hydroxy-2′,4′,6′-trimethyl-3′-((methyl(2,2,14-

trimethyl-4,7,10,13-tetraoxo-3-oxa-5,8,11,14-tetraazahexadecan-

16-yl)carbamoyl)oxy)-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl acetate

337
361
((2′S,3′R,6′R)-3′-((((2,5-dioxopyrrolidin-1-yl)oxy)carbonyl)oxy)-6′-

hydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl acetate

338
362
((2′S,3′R,6′R)-3′,6′-dihydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl

methanesulfonate

339
363
(2′S,3′R,6′R)-2′-(aminomethyl)-3′,6′-dihydroxy-2′,4′,6′-trimethyl-

2′,3′-dihydrospiro[cyclopropane-1,5′-inden]-7′(6′H)-one

340
364
(2′R,3′R,6′R)-3′-amino-6′-hydroxy-2′-(hydroxymethyl)-2′,4′,6′-

trimethyl-2′,3′-dihydrospiro[cyclopropane-1,5′-inden]-7′(6′H)-one

341
366
tert-butyl (R)-((6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)methyl)carbamate

342
367
tert-butyl (R)-(3-(6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-6′,7′-

dihydrospiro[cyclopropane-1,5′-inden]-3′-yl)propyl)carbamate

343
368
((2′S,3′R,6′R)-3′-acetoxy-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-

2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)

methyl acetate

344
369
(2′S,3′R,6′R)-2′-(((tert-butyldimethylsilyl)oxy)methyl)-3′,6′-

dihydroxy-2′,4′,6′-trimethyl-2′,3′-dihydrospiro[cyclopropane-1,5′-

inden]-7′(6′H)-one

345
370
(2′S,3′R,6′R)-2′-(((tert-butyldimethylsilyl)oxy)methyl)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-

1,5′-inden]-3′-yl acetate

346
371
(2′S,3′R,6′R)-6′-hydroxy-2′-(hydroxymethyl)-2′,4′,6′-trimethyl-7′-

oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl

acetate

347
372
(2′R,3′R,6′R)-2′-formyl-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-

2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl acetate

348
373
(2′5,3′R,6′R)-2′-(((((9H-fluoren-9-yl)methoxy)carbonyl)

amino)methyl)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydro spiro lcyclopropane-1,5′-inden]-3′-yl acetate

349
374
(2S,3′R,6′R)-2′-(amino methyl)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-

2′,3,6,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-3′-yl acetate

350
377
((2′S,3′R,6′R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-3′-

((tributylsilyl)oxy)-2′,3′,6′,7′-tetrahydrospiro lcyclopropane-1,5′-

inden]-2′-yl)methyl acetate

351
378
(2′S′R,6′R)-2′-(((tert-butoxycarbonyl)amino)methyl)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-2′,3,6′,7′-tetrahydrospiro[cyclopropane-

1,5′-inden]-3′-yl acetate

352
379
((2′S,3′R,6′R)-3′-(((2-((((4-((14S,17S)-1-(9H-fluoren-9-yl)-14-

isopropyl-3,6,9,12,15-pentaoxo-17-(3-ureidopropyl)-2-oxa-

4,7,10,13,16-pentaazaoctadecan-18-amido)benzyl)oxy)

carbonyl)(methyl)amino)ethyl)(methyl)carbamoyl)

oxy)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl acetate

353
380
((2′S′R,6′R)-3′-(((2-((((4-((2S,5S)-14-amino-5-isopropyl-

4,7,10,13-tetraoxo-2-(3-ureidopropyl)-3,6,9,12-

tetraazatetradecanamido)benzyl)oxy)carbonyl)(methyl)amino)

ethyl)(methyl)carbamoyl)oxy)-6′-hydroxy-2′,4′,6′-trimethyl-7′-

oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)

methyl acetate

354
381
(2′S,3′R,6′R)-6′-hydroxy-2′-(hydroxymethyl)-2′,4′,6′-trimethyl-3′-

((tributylsilyl)oxy)-2′,3′-dihydrospiro[cyclopropane-1,5′-inden]-

7′(6′H)-one

355
382
(2′R,3′R,6′R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-3′-

((tributylsilyl)oxy)-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-

indenel-2′-carbaldehyde

356
383
(2′R,3′R,6′R)-3′,6′-dihydroxy-2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-

tetrahydrospiro[cyclopropane-1,5′-indene]-2′-carbaldehyde

357
384
tert-butyl (((2′5,3′R,6′R)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-3′-

((tributylsilyl)oxy)-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-

inden]-2′-yl)methyl)carbamate

358
389
tert-butyl (((2′S,3′R,6′R)-3′,6′-dihydroxy-2′,4′,6′-trimethyl-7′-oxo-

2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-2′-

yl)methyl)carbamate

359
392
((2′S,3′S,6′R)-3′-(((2-(2-(2-(2-azidoacetamido)acetamido)-N-

methylacetamido)ethyl)(methyl)carbamoyl)oxy)-6′-hydroxy-

2′,4′,6′-trimethyl-7′-oxo-2′,3′,6′,7′-tetrahydrospiro[cyclopropane-

1,5′-inden]-2′-yl)methyl acetate

360
393
((2′S,3′R,6′R)-3′-(((2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-

yl)ethyl)carbamoyl)oxy)-6′-hydroxy-2′,4′,6′-trimethyl-7′-oxo-

2′,3′,6′,7′-tetrahydrospiro[cyclopropane-1,5′-inden]-2′-yl)methyl

acetate

361
394
(R)-6′-hydroxy-3′-((hydroxyamino)methyl)-2′,4′,6′-

trimethylspiro[cyclopropane-1,5′-inden]-7′(6′H)-one

TABLE 15

A summary NCI DTP 60 Cell Line showing Growth Inhibition (GI), Total Growth

Inhibition (TGI, the concentration of test drug where 100 × (T − T0)/(C − T0) = 0, i.e.

TGI signifies a cytostatic effect) and Lethal Concentration Dose (a cytotoxic effect, LD)

Data for selected Other Drugs, naturally occurring illudins, and Illudofulvene Analogs.

Mean GI50
Mean TGI
Mean LD50

NAME/NSC
inhibition
cytostatic
cytotoxic

Pyrrolobenzodiazepines 694501
7
nM
302
nM
>23,000
nM

Maytansine 153858
19
nM
318
nM
49,200
nM

Fumagillol 642492
6,130
nM
9,850
nM
>50,000
nM

Dolstatin-10376128
17
nM
2,680
nM
>50,000
nM

Auristatins 654663
1.4
nM
902
nM
>5,000
nM

Enadiyne 157365
2,900
nM
>100,000
nM
>100,000
nM

Halichondrin B 609395
1.2
nM
199
nM
>1,000
nM

Tubulysin A
12
nM
1,318
nM
>10,000
nM

Illudin S
10
nM
64
nM
511
nM

Illudin M
3
nM
20
nM
291
nM

Illudofulvene Analog 0016
102
nM
1,070
nM
4,270
nM

Illudofulvene Analog 0142
447
nM
4,667
nM
53,700
nM

Illudofulvene Analog 0159
117
nM
2,090
nM
14,790
nM

TABLE 16

A comparison of Illudofulvene Analog 002 growth inhibition values

obtained from cells in the NCI DTP 60 cell line panel to expression

of various DNA repair genes, to an embodiment of the invention.

GENE
GENE SEQ ID Nos
P value

ERCC2
21, 22, 145 through 155
P = 0.0246

ERCC8
1, 2, 35 through 45
P = 0.007

GTF2H1
5, 6, 57 through 67
P = 0.0007

PTGR1
249, 250, 251 through 261
P < 0.0001

USP7
1, 16, 112 through 122
P < 0.0001

XPC
none
P > 0.20

XRCC1
none
P > 0.20

RAD50
none
P > 0.20

TABLE 17

A comparison of Illudofulvene Analog 142 growth inhibition values obtained

from cells in the NCI DTP 60 cell line panel to expression of various

DNA repair genes, according to an embodiment of the invention.

GENE
GENE SEQ ID Nos
P value

ERCC2
21, 22, 145 through 155
P = 0.029

ERCC3
19, 20, 134 through 144
P = 0.0167

ERCC6
3, 4, 46 through 56
P = 0.0297

ERCC8
1,2, 35 through 45
P = 0.0159

PTGR1
249, 250, 251 through 261
P < 0.0001

USP7
1, 16, 112 through 122
P < 0.0001

XPC
none
P > 0.20

XRCC1
none
P > 0.20

RAD50
none
P > 0.20

TABLE 18

A comparison of Illudofulvene Analog 184 growth inhibition values

obtained from cells in the NCI DTP 60 cell line panel to expression of

various DNA repair genes,according to an embodiment of the invention.

GENE
GENE SEQ ID Nos
P value

ERCC3
19, 20, 134 through 144
P = 0.0252

ERCC8
1,2, 35 through 45
P = 0.0125

GTF2H2
5, 6, 68 through 78
P = 0.0107

PTGR1
249, 250, 251 through 261
P < 0.0001

USP7
15, 16, 112 through 122
P < 0.0001

MYC
222, 223, 224 through 235
P < 0.005

XPA
27, 28, 178 through 188
P = 0.0219

XPC
none
P > 0.20

XRCC1
none
P > 0.20

RAD50
none
P > 0.20

TABLE 19

A comparison of Illudofulvene Analog 176 growth inhibition values

obtained from cells in the NCI DTP 60 cell line panel to expression of

various DNA repair genes, according to an embodiment of the invention.

GENE
GENE SEQ ID Nos
P value

ERCC3
19,20, 134 through 144
P = 0.0258

ERCC8
1, 2, 35 through 45
P = 0.042

RAD18
29,30, 189 through 199
P = 0.0147

GTF2H3
9, 10, 79 through 89
P = 0.0037

GTF2H5
13,14 101 through 111
P = 0.0387

PTGR1
249, 250, 251 through 261
P < 0.0001

USP7
15, 16, 112 through 122
P < 0.0001

MYC
222, 223, 224 through 235
P < 0.002

XPC
none
P > 0.20

XRCC1
none
P > 0.20

RAD50
none
P > 0.20

TABLE 20

A comparison of the Illudin S growth inhibition

values obtained from cells in the NCI DTP 60 cell

line panel to expression of various DNA repair genes.

GENE
GENE SEQ ID Nos.
P value

ERCC3
19, 20, 134 through 144
P = 0.032

ERCC1
236, 237, 238 through 248
P = 0.0102

GTF2H1
5, 6, 57 through 67
P = 0.0003

PTGR1
249, 250, 251 through 261
P < 0.0244

PTGR2
262, 263, 264 through 274
P = 0.0382

PTGR1 + 2*
249 through 274
P = 0.0031

USP7
15, 16, 112 through 122
P < 0.0103

MYC
222, 223, 224 through 235
P < 0.0134

XPC
none
P > 0.20

XRCC1
none
P > 0.20

RAD50
none
P > 0.20

*PTGR1 + 2 refers to the sum of the activity of PTGR1 gene and the PTGR2 gene.

Analysis of Primary Tumor Cell Sensitivity.

Visible residues of healthy tissue are removed from the tumor sample excised from the patient or obtained from peripheral blood or other fluids. The tumor can be washed with PBS, placed in a petri dish and 2 ml of Primary Cancer Cell Medium D-ACF (PromoCell #C-2081) added and the tumor dissected into small pieces, then homogenized to 1 mm³particles. The tissue can be pelleted, washed, suspended then plated into NCDD-treated tissue culture vessels (PromoCell #C43080) containing Primary Cancer Cell Medium D-ACF. After 7 days the adherent cells are harvested, split, and plated into NCDD-treated tissue culture vessels (PromoCell #C43080) containing Primary Cancer Cell Medium D-ACF. A portion of the harvested adherent tumor cells are plated into 6 well tissue culture plates containing Primary Cancer Cell Medium D-ACF. Cells are allowed to adhere overnight. An illudofulvene is then added at: 0 ng/mL (control), 30 ng/mL, 100 ng/mL, 300 ng/mL, 1000 ng/mL, 3000 ng/mL. Cell viability at each concentration can then be determined and compared to the control population at 48 hours later by using a viability dye such as MTT, trypan blue or Sulforhodamine B. Tumor cell populations with an IC50 less than 300 ng/mL can be considered sensitive to the illudofulvene, whereas those with an IC50 greater than 3,000 ng/mL can be considered resistant. The illudofulvene can be administered to the patient based on the sensitivity. For example, in an embodiment of the invention, the tumor sample can be exposed to the desired Illudofulvene Analog under study at half-log internal concentrations from 1 ng/mL to 10,000 ng/mL, and cell viability determined using a viability dye (MTT, trypan blue, Sulforhodamine B, etc.) after 48 hours of exposure to the Illudofulvene Analog, and the IC50 determined. Tumor cells with an IC50 less than the median IC50 obtained for that Illudofulvene Analog in the NCI DTP 60 cell line panel can be considered sensitive whereas those with an IC50 greater than the median IC50 obtained for that Illudofulvene in the NCI DTP 60 cell line panel can be considered resistant.

Further Embodiments

Embodiments contemplated herein include Embodiments P1-P125 following.

Embodiment P1. An illudofulvene for use in the treatment of a cancer expressing an attenuated TCR biomarker, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with a device comprising single-stranded nucleic acid molecules capable of specifically hybridizing with the nucleotides of a TCR gene;

(b) detecting a level of expression of a TCR biomarker by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; and (c) determining that the TCR biomarker is attenuated.

Embodiment P2. The illudofulvene of Embodiment P1, where the TCR gene is selected from the group consisting of an ERCC gene, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment P3. The illudofulvene of Embodiment P2, where the ERCC gene is selected from the group consisting of ERCC1 gene, ERCC2 gene, ERCC3 gene, ERCC4 gene, ERCC5 gene, ERCC6 gene, ERCC6L gene, ERCC6L2 gene, and ERCC8 gene.

Embodiment P4. The illudofulvene of Embodiment P1, where the illudofulvene is an acylfulvene.

Embodiment P5. The illudofulvene of Embodiment P1, where the illudofulvene is an illudin.

Embodiment P6. An illudofulvene for use in a method of treating a patient with cancer, wherein the method includes: (a) determining whether a test sample from a patient has an attenuated TCR biomarker; and (b) if the test sample from the patient includes the attenuated TCR biomarker, then administer an effective amount of the illudofulvene.

Embodiment P7. The illudofulvene of Embodiment P6, where the effective amount is administered in a dosage between: a lower limit of approximately 0.1 mg of the illudofulvene per kilogram of body weight of the patient; and an upper limit of approximately 5 mg of the illudofulvene per kilogram of body weight of the patient.

Embodiment P8. The illudofulvene of Embodiment P6, where the test sample includes one or more nucleic acid molecules.

Embodiment P9. The illudofulvene of Embodiment P6, where the test sample hybridized specifically with the nucleotides of a TCR gene.

Embodiment P10. The illudofulvene of Embodiment P9, where the TCR gene is selected from the group consisting of ERCC gene, XPA gene, GTF series gene, UV gene, and USP gene.

Embodiment P11. The illudofulvene of Embodiment P6, where the TCR biomarker is determined to be attenuated by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the test sample.

Embodiment P12. The illudofulvene of Embodiment P6, where the illudofulvene is an acylfulvene.

Embodiment P13. The illudofulvene of Embodiment P6, where the illudofulvene is an illudin.

Embodiment P14. An illudofulvene for use in the treatment of a cancer expressing an enhanced MYC biomarker, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with a device comprising single-stranded nucleic acid molecules capable of specifically hybridizing with the nucleotides of a MYC gene; (b) detecting a level of expression of a MYC biomarker by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; and (c) determining that the MYC biomarker is enhanced.

Embodiment P15. The illudofulvene of Embodiment P14, further comprising where a PTGR biomarker is enhanced in the sample.

Embodiment P16. The illudofulvene of Embodiment P14, further comprising where a TCR biomarker is attenuated in the sample.

Embodiment P17. The illudofulvene of Embodiment P14, where the illudofulvene is an acylfulvene.

Embodiment P18. The illudofulvene of Embodiment P14, where the illudofulvene is an illudin.

Embodiment P19. An illudofulvene for use in a method of treating a patient with cancer, wherein the method includes: (a) determining whether a test sample from a patient has a MYC biomarker that is enhanced; and (b) if the test sample from the patient includes the enhanced MYC biomarker, then administer an effective amount of the illudofulvene.

Embodiment P20. The illudofulvene of Embodiment P18, where the effective amount is administered in a dosage between: a lower limit of approximately 0.1 mg of the illudofulvene per kilogram of body weight of the patient; and an upper limit of approximately 5 mg of the illudofulvene per kilogram of body weight of the patient.

Embodiment P21. The illudofulvene of Embodiment P19, where the test sample includes one or more nucleic acid molecules.

Embodiment P22. The illudofulvene of Embodiment P19, where the test sample hybridized specifically with the nucleotides of a MYC gene.

Embodiment P23. The illudofulvene of Embodiment P19, where the MYC biomarker is determined to be enhanced by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the test sample.

Embodiment P24. The illudofulvene of Embodiment P19, where the illudofulvene is an acylfulvene.

Embodiment P25. The illudofulvene of Embodiment P19, where the illudofulvene is an illudin.

Embodiment P26. The illudofulvene of Embodiment P19, further comprising where a PTGR biomarker is enhanced in the test sample.

Embodiment P27. The illudofulvene of Embodiment P26, further comprising where a TCR biomarker is attenuated in the test sample.

Embodiment P28. An illudofulvene for use in the treatment of a cancer expressing an enhanced MYC biomarker and an attenuated TCR biomarker, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with a device comprising single-stranded nucleic acid molecules capable of specifically hybridizing with the nucleotides of a TCR gene and a MYC gene; (b) detecting a level of expression of a TCR biomarker and a MYC biomarker by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; and (c) determining that the MYC biomarker is enhanced and the TCR biomarker is attenuated.

Embodiment P29. The illudofulvene of Embodiment P28, further comprising where a PTGR biomarker is enhanced in the sample.

Embodiment P30. The illudofulvene of Embodiment P28, where the illudofulvene is an acylfulvene.

Embodiment P31. The illudofulvene of Embodiment P28, where the illudofulvene is an illudin.

Embodiment P32. The illudofulvene of Embodiment P28, where the TCR gene is selected from the group consisting of ERCC gene, XPA gene, GTF series gene, UV gene, and USP gene.

Embodiment P33. The illudofulvene of Embodiment P28, where the TCR biomarker is attenuated due to truncating mutations, mutations abolishing enzyme activity, deletions or other chromosomal, transcription, translational and post translation event that interfering with TCR gene expression and/or subsequent TCR gene activity.

Embodiment P34. The illudofulvene of Embodiment P28, where the MYC biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P35. An illudofulvene for use in a method of treating a patient with cancer, wherein the method includes: (a) determining whether a test sample from a patient has an enhanced MYC biomarker; (b) determining whether the test sample from a patient has an attenuated TCR biomarker; (c) if the test sample from the patient includes the enhanced MYC biomarker and the attenuated TCR biomarker, then administer an effective amount of the illudofulvene.

Embodiment P36. The illudofulvene of Embodiment P35, where the effective amount is administered in a dosage between: a lower limit of approximately 0.1 mg of the illudofulvene per kilogram of body weight of the patient; and an upper limit of approximately 5 mg of the illudofulvene per kilogram of body weight of the patient.

Embodiment P37. The illudofulvene of Embodiment P35, where the test sample includes one or more nucleic acid molecules.

Embodiment P38. The illudofulvene of Embodiment P35, where the test sample hybridized specifically with the nucleotides of a MYC gene.

Embodiment P39. The illudofulvene of Embodiment P35, where the test sample hybridized specifically with the nucleotides of a TCR gene.

Embodiment P40. The illudofulvene of Embodiment P35, where the MYC biomarker is determined to be enhanced by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the test sample.

Embodiment P41. The illudofulvene of Embodiment P35, where the TCR biomarker is determined to be attenuated by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the test sample.

Embodiment P42. The illudofulvene of Embodiment P35, where the illudofulvene is an acylfulvene.

Embodiment P43. The illudofulvene of Embodiment P35, where the illudofulvene is an illudin.

Embodiment P44. The illudofulvene of Embodiment P35, further comprising where a PTGR biomarker is enhanced in the test sample.

Embodiment P45. The illudofulvene of Embodiment P35, further comprising where the TCR gene is selected from the group consisting of ERCC gene, XPA gene, GTF series gene, UV gene, and USP gene.

Embodiment P46. The illudofulvene of Embodiment P35, where the MYC biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P47. An illudofulvene for use in the treatment of a cancer expressing an enhanced PTGR biomarker, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with a device comprising single-stranded nucleic acid molecules capable of specifically hybridizing with the nucleotides of a PTGR gene; (b) detecting a level of expression of a PTGR biomarker by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; and (c) determining that the PTGR biomarker is attenuated.

Embodiment P48. The illudofulvene of Embodiment P47, where the PTGR gene is selected from the group consisting of PTGR1 and PTGR2.

Embodiment P49. The illudofulvene of Embodiment P47, where the illudofulvene is an acylfulvene.

Embodiment P50. The illudofulvene of Embodiment P47, where the illudofulvene is an illudin.

Embodiment P51. The illudofulvene of Embodiment P47, where the PTGR biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P52. An illudofulvene for use in a method of treating a patient with cancer, wherein the method includes: (a) determining whether a test sample from a patient has an enhanced PTGR biomarker; and (b) if the test sample from the patient includes the enhanced PTGR biomarker, then administer an effective amount of the illudofulvene.

Embodiment P53. The illudofulvene of Embodiment P52, where the effective amount is administered in a dosage between: a lower limit of approximately 0.1 mg of the illudofulvene per kilogram of body weight of the patient; and an upper limit of approximately 5 mg of the illudofulvene per kilogram of body weight of the patient.

Embodiment P54. The illudofulvene of Embodiment P53, where the test sample includes one or more nucleic acid molecules.

Embodiment P55. The illudofulvene of Embodiment P53, where the test sample hybridized specifically with the nucleotides of a PTGR gene.

Embodiment P56. The illudofulvene of Embodiment P53, where the PTGR gene is selected from the group consisting of PTGR1 and PTGR2.

Embodiment P57. The illudofulvene of Embodiment P53, where the PTGR biomarker is determined to be attenuated by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the test sample.

Embodiment P58. The illudofulvene of Embodiment P53, where the illudofulvene is an acylfulvene.

Embodiment P59. The illudofulvene of Embodiment P53, where the illudofulvene is an illudin.

Embodiment P60. The illudofulvene of Embodiment P53, where the PTGR gene includes PTGR1 and PTGR2.

Embodiment P61. The illudofulvene of Embodiment P53, where the PTGR biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P62. An illudofulvene for use in the treatment of a cancer expressing an enhanced MYC biomarker, an enhanced PTGR biomarker, and an attenuated TCR biomarker, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with a device comprising single-stranded nucleic acid molecules capable of specifically hybridizing with the nucleotides of a TCR gene, a PTGR gene, and a MYC gene; (b) detecting a level of expression of a TCR biomarker, a PTGR biomarker, and a MYC biomarker by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; and (c) determining that the MYC biomarker is enhanced, the PTGR biomarker is enhanced, and the TCR biomarker is attenuated.

Embodiment P63. The illudofulvene of Embodiment P62, where the PTGR gene is selected from the group consisting of PTGR1 and PTGR2.

Embodiment P64. The illudofulvene of Embodiment P62, where the TCR gene is selected from the group consisting of ERCC gene, XPA gene, GTF series gene, UV gene, and USP gene.

Embodiment P65. The illudofulvene of Embodiment P62, where the TCR biomarker is attenuated due to truncating mutations, mutations abolishing enzyme activity, deletions or other chromosomal, transcription, translational and post translation event that interfering with ERCC gene expression and subsequent gene activity.

Embodiment P66. The illudofulvene of Embodiment P62, where the PTGR biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P67. The illudofulvene of Embodiment P62, where the MYC biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P68. The illudofulvene of Embodiment P62, where the illudofulvene is an acylfulvene.

Embodiment P69. The illudofulvene of Embodiment P62, where the illudofulvene is an illudin.

Embodiment P70. An illudofulvene for use in a method of treating a patient with cancer, wherein the method includes: (a) determining whether a test sample from a patient has an enhanced MYC biomarker; (b) determining whether the test sample from a patient has an attenuated TCR biomarker; (c) determining whether the test sample from a patient has an enhanced PTGR biomarker; (d) if the test sample from the patient includes the enhanced MYC biomarker, the enhanced PTGR biomarker, and the attenuated TCR biomarker, then administer an effective amount of the illudofulvene.

Embodiment P71. The illudofulvene of Embodiment P70, where the effective amount is administered in a dosage between: a lower limit of approximately 0.1 mg of the illudofulvene per kilogram of body weight of the patient; and an upper limit of approximately 5 mg of the illudofulvene per kilogram of body weight of the patient.

Embodiment P72. The illudofulvene of Embodiment P70, where the test sample includes one or more nucleic acid molecules.

Embodiment P73. The illudofulvene of Embodiment P70, where the test sample hybridized specifically with the nucleotides of a MYC gene.

Embodiment P74. The illudofulvene of Embodiment P70, where the test sample hybridized specifically with the nucleotides of a TCR gene.

Embodiment P75. The illudofulvene of Embodiment P70, where the test sample hybridized specifically with the nucleotides of a PTGR gene.

Embodiment P76. The illudofulvene of Embodiment P70, where the MYC biomarker is determined to be enhanced by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the test sample.

Embodiment P77. The illudofulvene of Embodiment P70, where the TCR biomarker is determined to be attenuated by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the test sample.

Embodiment P78. The illudofulvene of Embodiment P70, where the illudofulvene is an acylfulvene.

Embodiment P79. The illudofulvene of Embodiment P70, where the illudofulvene is an illudin.

Embodiment P80. The illudofulvene of Embodiment P70, further comprising where the TCR gene is selected from the group consisting of ERCC gene, XPA gene, GTF series gene, UV gene, and USP gene.

Embodiment P81. The illudofulvene of Embodiment P70, where the PTGR gene is selected from the group consisting of PTGR1 and PTGR2.

Embodiment P82. The illudofulvene of Embodiment P70, where the MYC biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P83. The illudofulvene of Embodiment P70, where the TCR biomarker is attenuated due to truncating mutations, mutations abolishing enzyme activity, deletions or other chromosomal, transcription, translational and/or post translation event that interfering with TCR gene expression and/or subsequent TCR gene activity.

Embodiment P84. The illudofulvene of Embodiment P70, where the PTGR biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P85. An illudofulvene for use in the treatment of a cancer expressing an enhanced PTGR biomarker, and an attenuated TCR biomarker, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with a device comprising single-stranded nucleic acid molecules capable of specifically hybridizing with the nucleotides of a TCR gene, and a PTGR gene; (b) detecting a level of expression of a TCR biomarker and a PTGR biomarker by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; and (c) determining that the PTGR biomarker is enhanced, and the TCR biomarker is attenuated.

Embodiment P86. The illudofulvene of Embodiment P85, where the PTGR gene is selected from the group consisting of PTGR1 and PTGR2.

Embodiment P87. The illudofulvene of Embodiment P85, where the TCR gene is selected from the group consisting of ERCC gene, XPA gene, GTF series gene, UV gene, and USP gene.

Embodiment P88. The illudofulvene of Embodiment P85, where the TCR biomarker is attenuated due to truncating mutations, mutations abolishing enzyme activity, deletions or other chromosomal, transcription, translational and post translation event that interfering with ERCC gene expression and subsequent gene activity.

Embodiment P89. The illudofulvene of Embodiment P85, where the PTGR biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P90. The illudofulvene of Embodiment P85, where the illudofulvene is an acylfulvene.

Embodiment P91. The illudofulvene of Embodiment P85, where the illudofulvene is an illudin.

Embodiment P92. An illudofulvene for use in a method of treating a patient with cancer, wherein the method includes: (a) determining whether the test sample from a patient has an attenuated TCR biomarker; (b) determining whether the test sample from a patient has an enhanced PTGR biomarker; (c) if the test sample from the patient includes the enhanced PTGR biomarker, and the attenuated TCR biomarker, then administer an effective amount of the illudofulvene.

Embodiment P93. The illudofulvene of Embodiment P92, where the effective amount is administered in a dosage between: a lower limit of approximately 0.1 mg of the illudofulvene per kilogram of body weight of the patient; and an upper limit of approximately 5 mg of the illudofulvene per kilogram of body weight of the patient.

Embodiment P94. The illudofulvene of Embodiment P92, where the test sample includes one or more nucleic acid molecules.

Embodiment P95. The illudofulvene of Embodiment P92, where the test sample hybridized specifically with the nucleotides of a TCR gene.

Embodiment P96. The illudofulvene of Embodiment P92, where the test sample hybridized specifically with the nucleotides of a PTGR gene.

Embodiment P97. The illudofulvene of Embodiment P92, where the TCR biomarker is determined to be attenuated by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the test sample.

Embodiment P98. The illudofulvene of Embodiment P92, where the illudofulvene is an acylfulvene.

Embodiment P99. The illudofulvene of Embodiment P92, where the illudofulvene is an illudin.

Embodiment P100. The illudofulvene of Embodiment P92, further comprising where the TCR gene is selected from the group consisting of ERCC gene, XPA gene, GTF series gene, UV gene, and USP gene.

Embodiment P101. The illudofulvene of Embodiment P92, where the PTGR gene is selected from the group consisting of PTGR1 and PTGR2.

Embodiment P102. The illudofulvene of Embodiment P92, where the TCR biomarker is attenuated due to truncating mutations, mutations abolishing enzyme activity, deletions or other chromosomal, transcription, translational and/or post translation event that interfering with TCR gene expression and/or subsequent TCR gene activity.

Embodiment P103. The illudofulvene of Embodiment P92, where the PTGR biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P104. An illudofulvene for use in the treatment of a cancer expressing an enhanced MYC biomarker, and an enhanced PTGR biomarker, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with a device comprising single-stranded nucleic acid molecules capable of specifically hybridizing with the nucleotides of a PTGR gene, and a MYC gene; (b) detecting a level of expression of a PTGR biomarker and a MYC biomarker by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; and (c) determining that the MYC biomarker is enhanced, and the PTGR biomarker is enhanced.

Embodiment P105. The illudofulvene of Embodiment P104, where the PTGR gene is selected from the group consisting of PTGR1 and PTGR2.

Embodiment P106. The illudofulvene of Embodiment P104, where the PTGR biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P107. The illudofulvene of Embodiment P104, where the MYC biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P108. The illudofulvene of Embodiment P104, where the illudofulvene is an acylfulvene.

Embodiment P109. The illudofulvene of Embodiment P104, where the illudofulvene is an illudin.

Embodiment P110. An illudofulvene for use in a method of treating a patient with cancer, wherein the method includes: (a) determining whether a test sample from a patient has an enhanced MYC biomarker; (b) determining whether the test sample from a patient has an enhanced PTGR biomarker; (c) if the test sample from the patient includes the enhanced MYC biomarker, and the enhanced PTGR biomarker, then administer an effective amount of the illudofulvene.

Embodiment P111. The illudofulvene of Embodiment P110, where the effective amount is administered in a dosage between: a lower limit of approximately 0.1 mg of the illudofulvene per kilogram of body weight of the patient; and an upper limit of approximately 5 mg of the illudofulvene per kilogram of body weight of the patient.

Embodiment P112. The illudofulvene of Embodiment P110, where the test sample includes one or more nucleic acid molecules.

Embodiment P113. The illudofulvene of Embodiment P110, where the test sample hybridized specifically with the nucleotides of a MYC gene.

Embodiment P114. The illudofulvene of Embodiment P110, where the test sample hybridized specifically with the nucleotides of a PTGR gene.

Embodiment P115. The illudofulvene of Embodiment P110, where the MYC biomarker is determined to be enhanced by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the test sample.

Embodiment P116. The illudofulvene of Embodiment P110, where the illudofulvene is an acylfulvene.

Embodiment P117. The illudofulvene of Embodiment P110, where the illudofulvene is an illudin.

Embodiment P118. The illudofulvene of Embodiment P110, where the PTGR gene is selected from the group consisting of PTGR1 and PTGR2.

Embodiment P119. The illudofulvene of Embodiment P110, where the MYC biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P120. The illudofulvene of Embodiment P110, where the PTGR biomarker is enhanced when cells transform from a normal state to cancer cells, or when the cells are exposed to prior anticancer drug treatment and become upregulated as a response to the prior anticancer drug treatment.

Embodiment P121. An illudofulvene for use in the treatment of a cancer that is resistant to one or more Other Drugs, comprising: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with a device comprising single-stranded nucleic acid molecules capable of specifically hybridizing with the nucleotides selected from the group consisting of ERCC8 (SEQ ID NOS:1, 2, 35-45), ERCC6 (SEQ ID NOS:3, 4, 46-56), GTF2H1 (SEQ ID NOS:5, 6, 57-67), GTF2H2 (SEQ ID NOS:7, 8, 68-78), GTF2H3 (SEQ ID NOS:9, 10, 79-89), GTF2H4 (SEQ ID NOS:11, 12, 90-100), GTF2H5 (SEQ ID NOS:13, 14, 101-111), USP7 (SEQ ID NOS:15, 16, 112-122), UVSSA (SEQ ID NOS:17, 18, 123-133), ERCC3 (SEQ ID NOS:19, 20, 134-144), ERCC2 (SEQ ID NOS:21, 22, 145-155), ERCC4 (SEQ ID NOS:23, 24, 156-166), ERCC5 (SEQ ID NOS:25, 26, 167-177), XPA (SEQ ID NOS:27, 28, 178-188), RAD18 (SEQ ID NOS:29, 30, 189-199), ERCC6L (SEQ ID NOS:31, 32, 200-210), ERCC6L2 (SEQ ID NOS:33, 34, 211-221), MYC (SEQ ID NOS:222, 223, 224-235), ERCC1 (SEQ ID NOS:236, 237, 238-248), PTGR1 (SEQ ID NOS:249, 250, 251-261), and/or PTGR2 (SEQ ID NOS:262, 263, 264-274); (b) detecting a level of expression of a plurality of biomarkers selected from ERCC8 (SEQ ID NOS:1, 2, 35-45), ERCC6 (SEQ ID NOS:3, 4, 46-56), GTF2H1 (SEQ ID NOS:5, 6, 57-67), GTF2H2 (SEQ ID NOS:7, 8, 68-78), GTF2H3 (SEQ ID NOS:9, 10, 79-89), GTF2H4 (SEQ ID NOS:11, 12, 90-100), GTF2H5 (SEQ ID NOS:13, 14, 101-111), USP7 (SEQ ID NOS:15, 16, 112-122), UVSSA (SEQ ID NOS:17, 18, 123-133), ERCC3 (SEQ ID NOS:19, 20, 134-144), ERCC2 (SEQ ID NOS:21, 22, 145-155), ERCC4 (SEQ ID NOS:23, 24, 156-166), ERCC5 (SEQ ID NOS:25, 26, 167-177), XPA (SEQ ID NOS:27, 28, 178-188), RAD18 (SEQ ID NOS:29, 30, 189-199), ERCC6L (SEQ ID NOS:31, 32, 200-210), ERCC6L2 (SEQ ID NOS:33, 34, 211-221), MYC (SEQ ID NOS:222, 223, 224-235), ERCC1 (SEQ ID NOS:236, 237, 238-248), PTGR1 (SEQ ID NOS:249, 250, 251-261), and/or PTGR2 (SEQ ID NOS:262, 263, 264-274) by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample to generate a biomarker profile; and (c) determining to administer the illudofulvene based on the biomarker profile.

Embodiment P122. The illudofulvene of Embodiment P121, further comprising administering one or more additional therapies prior to, concurrently with, or after administration of the illudofulvene.

Embodiment P123. The illudofulvene of Embodiment P122, where one or more additional therapies includes surgery, radiation, or one or more Other Drugs.

Embodiment P124. The illudofulvene of Embodiment P121, where the cancer is selected from the group consisting of prostate cancer, ovarian cancer, hepatocellular carcinoma (HCC), cervical cancer, renal cell carcinoma (RCC), esophageal cancer, melanoma, glioma, pancreatic cancer, gastrointestinal stromal tumors (GIST), sarcoma, estrogen receptor-positive (ERpos) breast cancer, non-small cell lung carcinoma (NSCLC), colon cancer, bladder cancer, squamous cell carcinoma of the head and neck (SCCHN), acute myelogenous leukemia (AML), acute lympho-blastic leukemia (ALL), chronic lymphocytic leukemia (CLL), myelodysplastic syndrome (MDS), chronic myelogenous leukemia-chronic phase (CMLCP), diffuse large B-cell lymphoma (DLBCL), cutaneous T-cell lymphoma (CTCL), peripheral T-cell lymphoma (PTCL), and Hodgkin's lymphoma.

Embodiment P125. The illudofulvene of Embodiment P121, where the biomarker profile includes (c) calculating a difference score by subtracting the mean expression levels of one or more of the plurality of biomarkers from the mean expression levels of the one or more of the plurality of biomarkers of sensitivity.

Embodiments contemplated herein further include Embodiments Q1-Q54 following.

Embodiment Q1. An illudofulvene for use in the treatment of a cancer expressing an attenuated TCR biomarker, wherein said expression includes: (a) contacting a sample from a human patient with a means for determining the regulation of a PTGR gene, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity; (b) determining the level of regulation of the PTGR gene; (b) determining the level of regulation of the PTGR gene; (c) analyzing the sample from the human patient with a means for determining the regulation of a MYC gene, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) RT-PCR, (iv) TCR Tumor Mutation Panel and (v) primary tumor cell sensitivity; (d) determining the level of regulation of the MYC gene; (e) analyzing the sample from the human patient with a means for determining the regulation of three (3) or more TCR genes, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) RT-PCR, (iv) TCR Tumor Mutation Panel and (v) primary tumor cell sensitivity; where one or more of the following conditions are fulfilled: (I) the PTGR gene is upregulated, the MYC gene is upregulated, and at least one (1) of the three (3) or more TCR genes is downregulated;

(II) the PTGR gene is upregulated, the MYC gene is not upregulated, and at least one (1) of the three (3) or more TCR genes is downregulated; (III) the PTGR gene is not upregulated, the MYC gene is upregulated, and at least two (2) of the three (3) or more TCR genes is downregulated; and (IV) the PTGR gene is not upregulated, the MYC gene is not upregulated, and at least three (3) of the three (3) or more TCR genes is downregulated.

Embodiment Q2. The illudofulvene of Embodiment Q1, where the TCR gene is selected from the group consisting of an ERCC gene, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment Q3. The illudofulvene of Embodiment Q1, where the PTGR gene upregulated is PTGR1, where the process of analysis is quantification of gene mRNA expression by probe analysis, where the primers selected to hybridize with the one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:307-308.

Embodiment Q4. An illudofulvene for use in a method of treating a patient with cancer, wherein the method includes: (a) determining whether a test sample from a patient has an upregulated PTGR gene; (b) analyzing the sample with a means for determining that at least one TCR gene is downregulated, where the test sample is analyzed with one or more methods selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity; and (c) if the test sample from the patient includes the upregulated PTGR gene and at least one downregulated TCR gene, then administer an effective amount of the illudofulvene.

Embodiment Q5. The illudofulvene of Embodiment Q4, where the TCR gene is selected from the group consisting of an ERCC gene, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment Q6. The illudofulvene of Embodiment Q4, where the PTGR gene upregulated is PTGR1, where the process of analysis is quantification of gene mRNA expression by probe analysis, where the primers selected to hybridize with the one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:307-308.

Embodiment Q7. The illudofulvene of Embodiment Q4, where the PTGR gene upregulated is PTGR1, where the process of analysis is microarray analysis, where the primers selected to hybridize with the one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:251-261.

Embodiment Q8. The illudofulvene of Embodiment Q4, where the PTGR gene upregulated is PTGR1, where the process of analysis is RT-PCR, where one or both the primers and probe are selected to hybridize with the one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:897-899.

Embodiment Q9. The illudofulvene of Embodiment Q4, where the PTGR gene upregulated is PTGR1, where the process of analysis is TCR Tumor Mutation panel, where the primers selected to hybridize with the one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:682-696.

Embodiment Q10. The illudofulvene of Embodiment Q4, where the PTGR gene upregulated is PTGR1, where the process of analysis is primary tumor cell sensitivity.

Embodiment Q11. The illudofulvene of Embodiment Q4, where the PTGR gene upregulated is PTGR2, where the process of analysis is quantification of gene mRNA expression by probe analysis, where the primers selected to hybridize with the one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:309-310.

Embodiment Q12. The illudofulvene of Embodiment Q4, where the PTGR gene upregulated is PTGR2, where the process of analysis is microarray analysis, where the primers selected to hybridize with the one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:264-274.

Embodiment Q13. The illudofulvene of Embodiment Q4, where the PTGR gene upregulated is PTGR2, where the process of analysis is RT-PCR, where one or both the primers and probe are selected to hybridize with the one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:900-902.

Embodiment Q14. The illudofulvene of Embodiment Q4, where the PTGR gene upregulated is PTGR2, where the process of analysis is TCR Tumor Mutation panel, where the primers selected to hybridize with the one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO: 715-727.

Embodiment Q15. The illudofulvene of Embodiment Q4, where the PTGR gene upregulated is PTGR2, where the process of analysis is primary tumor cell sensitivity.

Embodiment Q16. The illudofulvene of Embodiment Q4, where the one or more TCR genes determined to be downregulated are analyzed using the TCR Tumor Mutation panel, where the primers selected to hybridize with the one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO: 329-440, and 451-851.

Embodiment Q17. An illudofulvene for use in treating cancer in a human patient in need thereof including administering the illudofulvene to the human patient, where the human patient has been determined to be responsive to the illudofulvene according to a process including: (a) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a PTGR gene; (b) detecting a level of expression of a PTGR mRNA by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (c) determining that the PTGR mRNA is upregulated in the human patient; (d) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a TCR gene; (e) detecting a level of expression of a plurality of TCR mRNAs by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (f) determining that the level of expression of at least one of the plurality of TCR mRNAs is downregulated in the human patient; and (g) administering the illudofulvene to the human patient, thereby treating cancer in the human patient.

Embodiment Q18. An illudofulvene for use in treating cancer in a human patient in need thereof including administering the illudofulvene to the human patient, where the human patient has been determined to be responsive to the illudofulvene according to a process including: (a) contacting a sample from the human patient including one or more nucleic acid molecules with a device including a plurality of primers capable of specifically hybridizing with nucleotides of a PTGR gene selected from the group consisting of SEQ ID NO:307-310; (b) detecting a level of expression of a PTGR mRNA by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (c) determining that the PTGR gene is upregulated in the human patient; (d) contacting a sample from the human patient including one or more nucleic acid molecules with a device including a plurality of primers capable of specifically hybridizing with nucleotides of a TCR gene selected from the group consisting of SEQ ID NO:275-304, and 311-322; (e) detecting a level of expression of a plurality of TCR mRNAs by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (f) determining that at least one of the plurality of TCR mRNAs is not upregulated in the human patient; and (g) administering the illudofulvene to the human patient, thereby treating cancer in the human patient.

Embodiment Q19. An illudofulvene for use in treating cancer in a human patient in need thereof including administering the illudofulvene to the human patient, where the human patient has been determined to be responsive to the illudofulvene according to a process including: (a) analyzing a sample from the human patient with a means for determining the regulation of a PTGR gene, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity; (b) determining that the PTGR gene is not upregulated; (c) analyzing a sample from the human patient with a means for determining the regulation of a MYC gene, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) RT-PCR, (iv) TCR Tumor Mutation Panel and (v) primary tumor cell sensitivity; (d) determining that the MYC gene is upregulated; (e) analyzing a sample from the human patient with a means for determining the regulation of a plurality of TCR genes, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) RT-PCR, (iv) TCR Tumor Mutation Panel and (v) primary tumor cell sensitivity; (f) determining that at least two of the plurality of TCR genes are downregulated; and (g) administering the illudofulvene to the human patient, thereby treating cancer in the human patient.

Embodiment Q20. The illudofulvene of Embodiment Q19, where the MYC gene determined to be upregulated is analyzed using the TCR Tumor Mutation panel, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:669-681.

Embodiment Q21. The illudofulvene of Embodiment Q19, where the process of analysis to determine that the PTGR gene is not upregulated is quantification of gene mRNA expression by probe analysis, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:307-310.

Embodiment Q22. The illudofulvene of Embodiment Q19, where the process of analysis to determine that the PTGR gene is not upregulated is microarray analysis, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:251-261, 264-274.

Embodiment Q23. The illudofulvene of Embodiment Q19, where the process of analysis to determine that the PTGR gene is not upregulated is RT-PCR, where one or both the primers and probe selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:897-902.

Embodiment Q24. The illudofulvene of Embodiment Q19, where the process of analysis to determine that the PTGR gene is not upregulated is TCR Tumor Mutation panel, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:682-696, 715-727.

Embodiment Q25. The illudofulvene of Embodiment Q19, where the process of analysis to determine that the PTGR gene is not upregulated is primary tumor cell sensitivity.

Embodiment Q26. The illudofulvene of Embodiment Q19, where the process of analysis to determine that one or more TCR genes are downregulated is quantification of gene mRNA expression by probe analysis, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:275-304 and 311-322.

Embodiment Q27. The illudofulvene of Embodiment Q19, where the process of analysis to determine that one or more TCR genes are downregulated is microarray analysis, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:35-248.

Embodiment Q28. The illudofulvene of Embodiment Q19, where the process of analysis to determine that one or more TCR genes are downregulated is RT-PCR, where one or both the primers and probe selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:852-893 and 903-917.

Embodiment Q29. The illudofulvene of Embodiment Q19, where the process of analysis to determine that one or more TCR genes are downregulated is TCR Tumor Mutation panel, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:329-440, and 451-851.

Embodiment Q30. The illudofulvene of Embodiment Q19, where the process of analysis to determine that one or more TCR genes are downregulated is primary tumor cell sensitivity.

Embodiment Q31. An illudofulvene for use in treating cancer in a human patient in need thereof including administering the illudofulvene to the human patient, where the human patient has been determined to be responsive to the illudofulvene according to a process including: (a) contacting a sample from the human patient including one or more nucleic acid molecules with one or more methods selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity to determine the level of expression of PTGR genes; (b) determining that the PTGR gene is not upregulated; (c) contacting a sample from the human patient including one or more nucleic acid molecules with one or more methods selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) RT-PCR, (iv) TCR Tumor Mutation Panel and (v) primary tumor cell sensitivity to determine the level of expression of a MYC gene; (d) determining that the MYC gene is upregulated; (e) contacting a sample from the human patient including one or more nucleic acid molecules with one or more methods selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) RT-PCR, (iv) TCR Tumor Mutation Panel and (v) primary tumor cell sensitivity to determine the level of expression of a plurality of TCR genes; (f) determining that the level of expression of at least two of the plurality of TCR genes are downregulated; and (g) administering the illudofulvene to the human patient, thereby treating cancer in the human patient.

Embodiment Q32. An illudofulvene for use in treating cancer in a human patient in need thereof including administering the illudofulvene to the human patient, where the human patient has been determined to be responsive to the illudofulvene according to a process including: (a) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a PTGR gene; (b) detecting a level of expression of a PTGR mRNA by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (c) determining that the level of expression of the PTGR mRNA is not upregulated in the human patient; (d) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a MYC gene; (e) detecting a level of expression of a MYC mRNA by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; and (f) determining that the level of expression of the MYC mRNA is upregulated in the human patient; (g) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a plurality of TCR genes; (h) detecting a level of expression of a plurality of TCR mRNAs by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (i) determining that the level of expression of at least two of the plurality of TCR mRNAs is downregulated in the human patient; and (j) administering the illudofulvene to the human patient, thereby treating cancer in the human patient.

Embodiment Q33. An illudofulvene for use in treating cancer in a human patient in need thereof including administering the illudofulvene to the human patient, where the human patient has been determined to be responsive to the illudofulvene according to a process including: (a) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a PTGR gene selected from the group consisting of SEQ ID NO:307-310; (b) detecting a level of expression of a PTGR mRNA by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (c) determining that the level of expression of the PTGR mRNA is NOT upregulated in the human patient; (d) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a MYC gene selected from the group consisting of SEQ ID NO:305-306; (e) detecting a level of expression of a MYC mRNA by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; and (f) determining that the level of expression of the MYC mRNA is upregulated in the human patient; (g) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a plurality of TCR genes selected from the group consisting of SEQ ID NO:275-304, and 311-322; (h) detecting a level of expression of a TCR mRNA by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (i) determining that the level of expression of at least two of the plurality of TCR mRNA is downregulated in the human patient; and (j) administering the illudofulvene to the human patient, thereby treating cancer in the human patient.

Embodiment Q34. An illudofulvene for use in treating cancer in a human patient in need thereof including administering the illudofulvene to the human patient, where the human patient has been determined to be responsive to the illudofulvene according to a process including: (a) analyzing a sample from the human patient with a means for determining the regulation of a PTGR gene, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity; (b) determining that the PTGR gene is not upregulated; (c) analyzing a sample from the human patient with a means for determining the regulation of a MYC gene, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) RT-PCR, (iv) TCR Tumor Mutation Panel and (v) primary tumor cell sensitivity; (d) determining that the MYC gene is not upregulated; (e) analyzing a sample from the human patient with a means for determining the regulation of three (3) or more TCR genes, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) RT-PCR, (iv) TCR Tumor Mutation Panel and (v) primary tumor cell sensitivity; (f) determining that at least three (3) of the three (3) or more TCR genes are downregulated; and (g) administering the illudofulvene to the human patient, thereby treating cancer in the human patient.

Embodiment Q35. The illudofulvene of Embodiment Q34, where the MYC gene determined to not be upregulated is analyzed using the TCR Tumor Mutation panel, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:669-681.

Embodiment Q36. The illudofulvene of Embodiment Q34, where the process of analysis to determine that the PTGR gene is not upregulated is quantification of gene mRNA expression by probe analysis, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:307-310.

Embodiment Q37. The illudofulvene of Embodiment Q34, where the process of analysis to determine that the PTGR gene is not upregulated is microarray analysis, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:251-261, 264-274.

Embodiment Q38. The illudofulvene of Embodiment Q34, where the process of analysis to determine that the PTGR gene is not upregulated is RT-PCR, where one or both the primers and probe selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:897-902.

Embodiment Q39. The illudofulvene of Embodiment Q34, where the process of analysis to determine that the PTGR gene is not upregulated is TCR Tumor Mutation panel, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:682-696, 715-727.

Embodiment Q40. The illudofulvene of Embodiment Q34, where the process of analysis to determine that the PTGR gene is not upregulated is primary tumor cell sensitivity.

Embodiment Q41. The illudofulvene of Embodiment Q34, where the process of analysis to determine that one or more TCR genes are downregulated is quantification of gene mRNA expression by probe analysis, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:275-304 and 311-322.

Embodiment Q42. The illudofulvene of Embodiment Q34, where the process of analysis to determine that one or more TCR genes are downregulated is microarray analysis, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:35-248.

Embodiment Q43. The illudofulvene of Embodiment Q34, where the process of analysis to determine that one or more TCR genes are downregulated is RT-PCR, where one or both the primers and probe selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:852-893 and 903-917.

Embodiment Q44. The illudofulvene of Embodiment Q34, where the process of analysis to determine that one or more TCR genes are downregulated is TCR Tumor Mutation panel, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:329-440, and 451-851.

Embodiment Q45. The illudofulvene of Embodiment Q34, where the process of analysis to determine that one or more TCR genes are downregulated is primary tumor cell sensitivity.

Embodiment Q46. An illudofulvene for use in treating cancer in a human patient in need thereof including administering the illudofulvene to the human patient, where the human patient has been determined to be responsive to the illudofulvene according to a process including: (a) contacting a sample from the human patient including one or more nucleic acid molecules with one or more methods selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity to determine the level of expression of PTGR genes; (b) determining that the level of expression of the PTGR gene is not upregulated; (c) contacting a sample from the human patient including one or more nucleic acid molecules with one or more methods selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) RT-PCR, (iv) TCR Tumor Mutation Panel and (v) primary tumor cell sensitivity to determine the level of expression of a MYC gene; (d) determining that the MYC gene is not upregulated; (e) contacting a sample from the human patient including one or more nucleic acid molecules with one or more methods selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) RT-PCR, (iv) TCR Tumor Mutation Panel and (v) primary tumor cell sensitivity to determine the level of expression of three (3) or more TCR genes; (f) determining that the level of expression of at least three (3) of the three (3) or more TCR genes are downregulated; and (g) administering the illudofulvene to the human patient, thereby treating cancer in the human patient.

Embodiment Q47. An illudofulvene for use in treating cancer in a human patient in need thereof including administering the illudofulvene to the human patient, where the human patient has been determined to be responsive to the illudofulvene according to a process including: (a) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a PTGR gene; (b) detecting a level of expression of a PTGR mRNA by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (c) determining that the level of expression of the PTGR mRNA is not upregulated in the human patient; (d) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a MYC gene; (e) detecting a level of expression of a MYC mRNA by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; and (f) determining that the level of expression of the MYC mRNA is not upregulated in the human patient; (g) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a plurality of TCR genes; (h) detecting a level of expression of three (3) or more TCR mRNAs by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (i) determining that the level of expression of at least three (3) of the three (3) or more TCR mRNAs is downregulated in the human patient; and (j) administering the illudofulvene to the human patient, thereby treating cancer in the human patient.

Embodiment Q48. An illudofulvene for use in treating cancer in a human patient in need thereof including administering the illudofulvene to the human patient, where the human patient has been determined to be responsive to the illudofulvene according to a process including: (a) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a PTGR gene selected from the group consisting of SEQ ID NO:307-310; (b) detecting a level of expression of a PTGR mRNA by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (c) determining that the level of expression of the PTGR mRNA is not upregulated in the human patient; (d) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a MYC gene selected from the group consisting of SEQ ID NO:305-306; (e) detecting a level of expression of a MYC mRNA by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; and (f) determining that the level of expression of the MYC mRNA is not upregulated in the human patient; (g) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a plurality of TCR genes selected from the group consisting of SEQ ID NO:275-304, and 311-322; (h) detecting a level of expression of three (3) or more TCR mRNAs by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (i) determining that the level of expression of at least three (3) of the three (3) or more TCR mRNAs is downregulated in the human patient; and (j) administering the illudofulvene to the human patient, thereby treating cancer in the human patient.

Embodiment Q49. An illudofulvene for use in treating cancer in a human patient in need thereof including administering the illudofulvene to the human patient, where the human patient has been determined to be responsive to the illudofulvene according to a process including: (a) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a PTGR gene; (b) detecting a level of expression of a PTGR mRNA by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (c) determining that the level of expression of the PTGR mRNA is not upregulated in the human patient; (d) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of a MYC gene; (e) detecting a level of expression of a MYC mRNA by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; (f) determining that the level of expression of the MYC mRNA is not upregulated in the human patient; (g) contacting a sample from the human patient including one or more nucleic acid molecules with a device including single-stranded nucleic acid molecules capable of specifically hybridizing with nucleotides of three (3) or more TCR genes; (h) detecting a level of expression of at least three (3) of the three (3) or more TCR mRNAs by performing microarray analysis or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) of the sample; and (i) determining that at least three (3) of the three (3) or more TCR mRNAs are upregulated in the human patient.

Embodiment Q50. An illudofulvene for use in treating cancer in a human patient in need thereof including administering the illudofulvene to the human patient, where the human patient has been determined to be responsive to the illudofulvene according to a process including: (a) analyzing a sample from the human patient with a means for determining that a PTGR gene is upregulated; (b) analyzing a sample from the human patient with a means for determining that at least one of the TCR genes is downregulated; and (c) administering the illudofulvene to the human patient, thereby treating cancer in the human patient.

Embodiment Q51. The illudofulvene of Embodiment Q19, where the MYC gene determined to be upregulated is not upregulated is RT-PCR, where one or both the primers and probe selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:894-896.

Embodiment Q52. The illudofulvene of Embodiment Q19, where the MYC gene determined to be upregulated is microarray analysis, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:224-235.

Embodiment Q53. The illudofulvene of Embodiment Q19, where the process of analysis to determine that the MYC gene is upregulated is primary tumor cell sensitivity.

Embodiment Q54. The illudofulvene of Embodiment Q19, where the MYC gene determined to be upregulated is quantification of gene mRNA expression by probe analysis, where the primers selected to hybridize with one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:305-306.

Embodiments contemplated herein further include Embodiments R1-R82 following.

Embodiment R1. An illudofulvene for use in the treatment of a cancer expressing an upregulated PTGR gene and an upregulated MYC gene, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with one or more means for determining the regulation of expression of one or more PTGR genes; (b) contacting the sample from the patient with one or more means for determining the regulation of expression of a MYC gene; (c) contacting the sample from the patient with one or more means for determining the regulation of expression of one or more TCR genes; (d) detecting a level of expression of the one or more PTGR genes that are upregulated; (e) detecting a level of expression of the MYC gene that is upregulated; and (f) detecting a deficiency in a level of expression of one or more TCR genes.

Embodiment R2. The illudofulvene of Embodiment R1, where the one or more TCR genes are selected from the group consisting of an ERCC gene, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment R3. The illudofulvene of Embodiments R1 or R2, where the one or more PTGR genes are selected from the group consisting of PTGR1 and PTGR2.

Embodiment R4. The illudofulvene of Embodiments R1-R3, where the one or more PTGR genes include PTGR1.

Embodiment R5. The illudofulvene of Embodiments R1-R4, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R6. The illudofulvene of Embodiments R1-R5, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R7. The illudofulvene of Embodiments R1-R3, where the one or more PTGR genes include PTGR2.

Embodiment R8. The illudofulvene of Embodiments R1-R3 or R7, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R9. The illudofulvene of Embodiments R1-R3 or R7-R8, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R10. An illudofulvene for use in a method of treating a patient with cancer, wherein the method includes: (a) determining whether a test sample from a patient has a level of expression of one or more PTGR genes that are upregulated, a level of expression of a MYC gene that is upregulated, and a deficiency in a level of expression of one or more TCR genes; and (b) if the test sample from the patient includes the one or more upregulated PTGR genes, the upregulated MYC gene, and the deficiency in the level of expression of one or more TCR genes, then administer an effective amount of the illudofulvene.

Embodiment R11. The illudofulvene of Embodiment R10, where the one or more TCR genes are selected from the group consisting of an ERCC gene, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment R12. The illudofulvene of Embodiments R10 or R11, where the one or more PTGR genes are selected from the group consisting of PTGR1 and PTGR2.

Embodiment R13. The illudofulvene of Embodiments R10-R12, where the means is where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R14. The illudofulvene of Embodiments R10-R13, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R15. The illudofulvene of Embodiments R10-R14, where the effective amount is administered in a dosage between: a lower limit of approximately 0.1 mg of the illudofulvene per kilogram of body weight of the patient; and an upper limit of approximately 5 mg of the illudofulvene per kilogram of body weight of the patient.

Embodiment R16. An illudofulvene for use in the treatment of a cancer expressing a PTGR gene and one or more TCR genes, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with one or more means for determining the regulation of expression of one or more PTGR genes; (b) contacting the sample from the patient with one or more means for determining the regulation of expression of one or more TCR genes; (c) detecting a level of expression of the one or more PTGR genes that are upregulated; and (d) detecting a deficiency in a level of expression of one or more TCR genes.

Embodiment R17. The illudofulvene of Embodiment R16, where the one or more TCR genes are selected from the group consisting of an ERCC gene, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment R18. The illudofulvene of Embodiments R16 or R17, where the one or more PTGR genes are selected from the group consisting of PTGR1 and PTGR2.

Embodiment R19. The illudofulvene of Embodiments R16-R8, where the one or more PTGR genes include PTGR1.

Embodiment R20. The illudofulvene of Embodiments R16-R19, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R21. The illudofulvene of Embodiments R16-R20, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R22. The illudofulvene of Embodiments R16-R18, where the one or more PTGR genes include PTGR2.

Embodiment R23. The illudofulvene of Embodiments R16-R18 or R22, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R24. The illudofulvene of Embodiments R16-R18 or R22-R23, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R25. An illudofulvene for use in the treatment of a cancer expressing a MYC gene and one or more TCR genes, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with one or more means for determining the regulation of expression of a MYC gene; (b) contacting the sample from the patient with one or more means for determining the regulation of expression of one or more TCR genes; (c) detecting a level of expression of the MYC gene that is upregulated; and (d) detecting a deficiency in a level of expression of one or more TCR genes.

Embodiment R26. The illudofulvene of Embodiment R25, where the one or more TCR genes are selected from the group consisting of an ERCC gene, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment R27. The illudofulvene of Embodiments R25 or R26, where the one or more PTGR genes are selected from the group consisting of PTGR1 and PTGR2.

Embodiment R28. The illudofulvene of Embodiments R25-R27, where the one or more PTGR genes include PTGR1.

Embodiment R29. The illudofulvene of Embodiments R25-R28, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R30. The illudofulvene of Embodiments R25-R29, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R31. The illudofulvene of Embodiments R25-R27, where the one or more PTGR genes include PTGR2.

Embodiment R32. The illudofulvene of Embodiments R25-R27 or R31, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R33. The illudofulvene of Embodiments R25-R27 or R31-R32, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R34. An illudofulvene for use in the treatment of a cancer expressing a PTGR gene, a MYC gene and one or more TCR genes, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with one or more means for determining the regulation of expression of one or more PTGR genes; (b) contacting the sample from the patient with one or more means for determining the regulation of expression of a MYC gene; (c) contacting the sample from the patient with one or more means for determining the regulation of expression of one or more TCR genes; (d) detecting either a level of expression of the one or more PTGR genes that are upregulated, or detecting a level of expression of the MYC gene that is upregulated; and (e) detecting a deficiency in a level of expression of one or more TCR genes.

Embodiment R35. The illudofulvene of Embodiment R34, where the one or more TCR genes are selected from the group consisting of one or more ERCC genes, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment R36. The illudofulvene of Embodiments R34 or R35, where the one or more PTGR genes are selected from the group consisting of PTGR1 and PTGR2.

Embodiment R37. The illudofulvene of Embodiments R34-R36, where the one or more PTGR genes include PTGR1.

Embodiment R38. The illudofulvene of Embodiments R34-R37, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R39. The illudofulvene of Embodiments R34-R38, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R40. The illudofulvene of Embodiments R34-R37, where the one or more PTGR genes include PTGR2.

Embodiment R41. The illudofulvene of Embodiments R34-R37 or R40, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R42. The illudofulvene of Embodiments R34-R37 or R40-R41, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R43. An illudofulvene for use in the treatment of a cancer expressing three or more TCR genes, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with one or more means for determining the regulation of expression of three or more TCR genes; and (b) detecting a deficiency in a level of expression of three or more TCR genes.

Embodiment R44. The illudofulvene of Embodiment R43, where the three or more TCR genes are selected from the group consisting of one or more ERCC genes, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment R45. The illudofulvene of Embodiments R43 or R44, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R46. An illudofulvene for use in the treatment of a cancer expressing a PTGR gene, a MYC gene and one or more TCR genes, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with one or more means for determining the regulation of expression of one or more PTGR genes; (b) contacting the sample from the patient with one or more means for determining the regulation of expression of a MYC gene; (c) contacting the sample from the patient with one or more means for determining the regulation of expression of one or more TCR genes; (d) detecting either a level of expression of the one or more PTGR genes that are upregulated, or detecting a level of expression of the MYC gene that is upregulated; (e) if one or more PTGR genes are upregulated then detecting a deficiency in a level of expression of one or more TCR genes; and (f) if the MYC gene is upregulated then detecting a deficiency in a level of expression of two or more TCR genes.

Embodiment R47. The illudofulvene of Embodiment R45, where the one or more TCR genes are selected from the group consisting of one or more ERCC genes, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment R48. The illudofulvene of Embodiment R46 or R47, where the one or more PTGR genes are selected from the group consisting of PTGR1 and PTGR2.

Embodiment R49. The illudofulvene of Embodiment R46 to R48, where the one or more PTGR genes include PTGR1.

Embodiment R50. The illudofulvene of Embodiment R46 to R49, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R51. The illudofulvene of Embodiment R46 to R50, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R52. The illudofulvene of Embodiment R46 to R48, where the one or more PTGR genes include PTGR2.

Embodiment R53. The illudofulvene of Embodiment R46 to R48 or R52, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R54. The illudofulvene of Embodiment R46 to R48 or R52 to 53, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R55. An illudofulvene for use in a method of treating a patient with cancer, wherein the method includes: (a) determining whether a test sample from a patient has a level of expression of one or more PTGR genes that are upregulated, a level of expression of a MYC gene that is upregulated, and a deficiency in a level of expression of one or more TCR genes; (b) if the test sample from the patient includes the one or more upregulated PTGR genes, and the deficiency in the level of expression of one or more TCR genes, then administer an effective amount of the illudofulvene; or (c) if the test sample from the patient includes the upregulated MYC gene, and the deficiency in the level of expression of two or more TCR genes, then administer an effective amount of the illudofulvene.

Embodiment R56. The illudofulvene of Embodiment R55, where the one or more TCR genes are selected from the group consisting of one or more ERCC genes, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment R57. The illudofulvene of Embodiment R55 or R56, where the one or more PTGR genes are selected from the group consisting of PTGR1 and PTGR2.

Embodiment R58. The illudofulvene of Embodiment R55 to R57, where the means is where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R59. The illudofulvene of Embodiment R55 to R58, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R60. The illudofulvene of Embodiment R55 to R59, where the effective amount is administered in a dosage between: a lower limit of approximately 0.1 mg of the illudofulvene per kilogram of body weight of the patient; and an upper limit of approximately 5 mg of the illudofulvene per kilogram of body weight of the patient.

Embodiment R61. An illudofulvene analog for use in the treatment of a cancerous tumor expressing a PTGR gene, a MYC gene or one or more TCR genes, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with one or more means for determining the regulation of expression of one or more PTGR genes, the regulation of expression of a MYC gene, and the regulation of expression of one or more TCR genes; and (b) determining if a cancerous tumor is sensitive to an illudofulvene analog based on a first relative ratio of expression of the one or more PTGR gene compared with a housekeeping gene and a second relative ratio of expression of the TCR genes compared with a housekeeping gene, or a third relative ratio of expression of the MYC gene compared with a housekeeping gene and the second relative ratio of expression.

Embodiment R62. The illudofulvene of Embodiment R61, where the one or more TCR genes are selected from the group consisting of one or more ERCC genes, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment R63. The illudofulvene of Embodiment R61 or 62, where the one or more PTGR genes are selected from the group consisting of PTGR1 and PTGR2.

Embodiment R64. The illudofulvene of Embodiment R61 to 63, where the one or more housekeeping genes are selecting from the group consisting of beta actin (ACTB), transferrin receptor (TFRC) and glyceraldehyde-3-phosphate dehydrogenase (GAPD).

Embodiment R65. The illudofulvene of Embodiment R61 to 64, where the one or more PTGR genes include PTGR1.

Embodiment R66. The illudofulvene of Embodiment R61 to 65, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R67. The illudofulvene of Embodiment R61 to 66, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R68. The illudofulvene of Embodiment R61 to 64, where the one or more PTGR genes include PTGR2.

Embodiment R69. The illudofulvene of Embodiment R61 to 44 or 68, where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R70. An illudofulvene for use in a method of treating a patient with cancer, wherein the method includes: (a) determining from a patient test sample a first relative ratio of expression of the one or more PTGR genes compared with at least one of the one or more housekeeping genes or determining a second relative ratio of expression of the MYC gene compared with at least one of the one or more housekeeping genes; (b) determining a third relative ratio of expression of the TCR genes compared with at least one of the one or more housekeeping genes; (c) if the first relative ratio of expression is high and the third relative ratio of expression is deficient or the second relative ratio of expression is high and the third relative ratio of expression is deficient, then administer an effective amount of the illudofulvene.

Embodiment R71. The illudofulvene of Embodiment R70, where the one or more TCR genes are selected from the group consisting of one or more ERCC genes, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment R72. The illudofulvene of Embodiment R70 or R71, where the one or more PTGR genes are selected from the group consisting of PTGR1 and PTGR2.

Embodiment R73. The illudofulvene of Embodiment R70 to R72, where the means is where the means is selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment R74. The illudofulvene of Embodiment R70 to R73, where one or more primers selected to hybridize with the one or more nucleic acid molecules used to quantify expression of one or more PTGR genes, the MYC gene and the one or more TCR genes are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, and SEQ ID NO:900-902.

Embodiment R75. The illudofulvene of Embodiment R70 to R74, where the effective amount is administered in a dosage between: a lower limit of approximately 0.1 mg of the illudofulvene per kilogram of body weight of the patient; and an upper limit of approximately 5 mg of the illudofulvene per kilogram of body weight of the patient.

Embodiment R76. An illudofulvene analog selected from the group consisting of illudin ring derivatives, illudin alkyl derivatives and acyl-fulvenes for use in the treatment of a cancerous tumor expressing a PTGR gene, a MYC gene or one or more TCR genes, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with one or more means for determining: (i) the regulation of expression of one or more PTGR genes; (ii) the regulation of expression of at least one TCR gene; (iii) the regulation of expression of at least one housekeeping gene; (b) determining if a cancerous tumor is sensitive to an illudofulvene analog based on a first relative ratio of expression of the one or more PTGR genes compared with the at least one housekeeping gene, when the first relative ratio of expression is high, and a second relative ratio of expression of the at least one TCR gene compared with the at least one housekeeping gene, when the second relative ratio of expression is deficient; or (c) contacting the sample comprising one or more nucleic acid molecules with one or more means for determining: (iv) the regulation of expression of a MYC gene; (v) the regulation of expression of two or more TCR genes; (vi) the regulation of expression of one or more housekeeping genes; (d) determining if the cancerous tumor is sensitive to the illudofulvene analog based on a third relative ratio of expression of the MYC gene compared with the one or more housekeeping genes, when the third relative ratio of expression is high, and a fourth relative ratio of expression of two or more TCR genes compared with the one or more housekeeping genes, when the fourth relative ratio of expression is deficient.

Embodiment R77. An illudofulvene analog selected from the group consisting of illudin ring derivatives and illudin alkyl derivatives for use in the treatment of a cancerous tumor expressing a PTGR gene, a MYC gene or one or more TCR genes, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with one or more means for determining: (i) the regulation of expression of one or more PTGR genes; (ii) the regulation of expression of at least one TCR gene; (iii) the regulation of expression of at least one housekeeping gene; (b) determining if a cancerous tumor is sensitive to an illudofulvene analog based on a first relative ratio of expression of the one or more PTGR genes compared with the at least one housekeeping gene, when the first relative ratio of expression is high, and a second relative ratio of expression of the at least one TCR gene compared with the at least one housekeeping gene, when the second relative ratio of expression is deficient; or (c) contacting the sample comprising one or more nucleic acid molecules with one or more means for determining: (iv) the regulation of expression of a MYC gene; (v) the regulation of expression of two or more TCR genes; (vi) the regulation of expression of one or more housekeeping genes; (d) determining if the cancerous tumor is sensitive to the illudofulvene analog based on a third relative ratio of expression of the MYC gene compared with the one or more housekeeping genes, when the third relative ratio of expression is high, and a fourth relative ratio of expression of two or more TCR genes compared with the one or more housekeeping genes, when the fourth relative ratio of expression is deficient.

Embodiment R78. An illudofulvene analog selected from the group consisting of illudins and acyl-fulvenes for use in the treatment of a cancerous tumor expressing a PTGR gene, a MYC gene or one or more TCR genes, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with one or more means for determining: (i) the regulation of expression of one or more PTGR genes; (ii) the regulation of expression of at least one TCR gene; (iii) the regulation of expression of at least one housekeeping gene; (b) determining if a cancerous tumor is sensitive to an illudofulvene analog based on a first relative ratio of expression of the one or more PTGR genes compared with the at least one housekeeping gene, when the first relative ratio of expression is high, and a second relative ratio of expression of the at least one TCR gene compared with the at least one housekeeping gene, when the second relative ratio of expression is deficient; or (c) contacting the sample comprising one or more nucleic acid molecules with one or more means for determining: (iv) the regulation of expression of a MYC gene; (v) the regulation of expression of two or more TCR genes; (vi) the regulation of expression of one or more housekeeping genes; (d) determining if the cancerous tumor is sensitive to the illudofulvene analog based on a third relative ratio of expression of the MYC gene compared with the one or more housekeeping genes, when the third relative ratio of expression is high, and a fourth relative ratio of expression of two or more TCR genes compared with the one or more housekeeping genes, when the fourth relative ratio of expression is deficient.

Embodiment R79. An illudofulvene analog selected from the group consisting of illudin ring derivatives, illudin alkyl derivatives and acyl-fulvenes for use in a method of treating a patient with cancer, wherein the method includes: (a) determining from a patient test sample a first relative ratio of expression based on expression of the one or more PTGR genes compared with expression of at least one housekeeping gene; (b) determining from the patient test sample a second relative ratio of expression based on expression of at least one TCR genes compared with expression of at least one housekeeping gene; (c) determining that the patient is sensitive, if the first relative ratio of expression is high and the second relative ratio of expression is deficient, then administer an effective amount of the illudofulvene; or (d) determining from a patient test sample a third relative ratio of expression based on expression of a MYC gene compared with at least one of the housekeeping genes; (e) determining from a patient test sample a fourth relative ratio of expression based on expression of two or more TCR genes compared with at least one of the housekeeping genes; (f) determining that the patient is sensitive, if the third relative ratio of expression is high and the fourth relative ratio of expression is deficient, then administer an effective amount of the illudofulvene.

Embodiment R80. An illudofulvene analog selected from the group consisting of illudin ring derivatives and illudin alkyl derivatives for use in a method of treating a patient with cancer, wherein the method includes: (a) determining from a patient test sample a first relative ratio of expression based on expression of the one or more PTGR genes compared with expression of at least one housekeeping gene; (b) determining from the patient test sample a second relative ratio of expression based on expression of at least one TCR genes compared with expression of at least one housekeeping gene; (c) determining that the patient is sensitive, if the first relative ratio of expression is high and the second relative ratio of expression is deficient, then administer an effective amount of the illudofulvene; or (d) determining from a patient test sample a third relative ratio of expression based on expression of a MYC gene compared with at least one of the housekeeping genes; (e) determining from a patient test sample a fourth relative ratio of expression based on expression of two or more TCR genes compared with at least one of the housekeeping genes; (f) determining that the patient is sensitive, if the third relative ratio of expression is high and the fourth relative ratio of expression is deficient, then administer an effective amount of the illudofulvene.

Embodiment R81. An illudofulvene analog selected from the group consisting of illudins and acyl-fulvenes for use in a method of treating a patient with cancer, wherein the method includes: (a) determining from a patient test sample a first relative ratio of expression based on expression of the one or more PTGR genes compared with expression of at least one housekeeping gene; (b) determining from the patient test sample a second relative ratio of expression based on expression of at least one TCR genes compared with expression of at least one housekeeping gene; (c) determining that the patient is sensitive, if the first relative ratio of expression is high and the second relative ratio of expression is deficient, then administer an effective amount of the illudofulvene; or (d) determining from a patient test sample a third relative ratio of expression based on expression of a MYC gene compared with at least one of the housekeeping genes; (e) determining from a patient test sample a fourth relative ratio of expression based on expression of two or more TCR genes compared with at least one of the housekeeping genes; (f) determining that the patient is sensitive, if the third relative ratio of expression is high and the fourth relative ratio of expression is deficient, then administer an effective amount of the illudofulvene.

Embodiment R82. An illudofulvene analog selected from the group consisting of illudin ring derivatives, illudin alkyl derivatives and acyl-fulvenes for in the treatment of a cancerous tumor expressing a PTGR gene, a MYC gene or one or more TCR genes, wherein said expression includes: (a) contacting a sample from a patient comprising one or more nucleic acid molecules with one or more means for determining: (i) the regulation of expression of one or more PTGR genes; (ii) the regulation of expression of at least one TCR gene; (iii) the regulation of expression of at least one housekeeping gene; (b) determining if a cancerous tumor is sensitive to an illudofulvene analog based on a first relative ratio of expression of the one or more PTGR genes compared with the at least one housekeeping gene, when the first relative ratio of expression is high, and a second relative ratio of expression of the at least one TCR gene compared with the at least one housekeeping gene, when the second relative ratio of expression is deficient; or (c) contacting the sample comprising one or more nucleic acid molecules with one or more means for determining: (iv) the regulation of expression of a MYC gene; (v) the regulation of expression of two or more TCR genes; (vi) the regulation of expression of one or more housekeeping genes; (d) determining if the cancerous tumor is sensitive to the illudofulvene analog based on a third relative ratio of expression of the MYC gene compared with the one or more housekeeping genes, when the third relative ratio of expression is high, and a fourth relative ratio of expression of two or more TCR genes compared with the one or more housekeeping genes, when the fourth relative ratio of expression is deficient; or (e) contacting the sample comprising one or more nucleic acid molecules with one or more means for determining: (vii) the regulation of expression of three or more TCR genes; (viii) the regulation of expression of one or more housekeeping genes; (f) determining if the cancerous tumor is sensitive to the illudofulvene analog based on a fifth relative ratio of expression of the three or more TCR genes compared with the one or more housekeeping genes.

Embodiments contemplated herein further include Embodiments S1-S21 following.

Embodiment S1. An illudofulvene analog for use in the treatment of a cancerous tumor expressing a PTGR gene, a MYC gene or one or more TCR genes in a patient, wherein said expression includes (a) increased expression of one or more PTGR genes compared to a housekeeping gene in the cancerous tumor; (b) increased expression of a MYC gene compared to a housekeeping gene in the cancerous tumor; and/or (c) decreased expression of one or more TCR genes compared with a housekeeping gene.

Embodiment S2. The illudofulvene analog of Embodiment S1, where the illudofulvene is selected from the group consisting of 002, 008, 012, 039, 040, 41, 42, 43, 44, 46, 75, 76, 77, 78, 79, 88, 90, 94, 103, 104, 108, 110, 112, 113, 114, 115, 116, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 130, 141, 142, 146, 149, 150, 160, 164, 165, 177, 178, 184. 189, 190, 196, 198, 199, 201, 202, 203, 207, 208, 209, 210, 217, 221, 223, 233, 234, 235, 240, 244, 254, 255, 259, 262, 263, 266, 267, 268, 269, 275, 276, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 304, 305, 306, 310, 314, 315, 317, 321, 323, 324, 332, 333, 334, 335, 336, 337, 338, 339, 340, 345, 346, 347, 348, 351, 352, 354, 356, 357, 368, 359, 361, 362, 363, 364, 365, 366, 367, 369, 370, 371, 372, 373, 374, 375, 377, 378, 379, 380, 381, 382, 383, 384, 389, 392, 393, and 394.

Embodiment S3. The illudofulvene analog of Embodiment S1 or S2, where the one or more housekeeping genes are selecting from the group consisting of beta actin (ACTB), transferrin receptor (TFRC) and glyceraldehyde-3-phosphate dehydrogenase (GAPD) and ubiquitin.

Embodiment S4. An illudofulvene analog for use in the treatment of a cancerous tumor expressing a PTGR gene, a MYC gene or one or more TCR genes in a patient, wherein said expression includes (a) messenger RNA and/or gene product (protein) of one or more PTGR genes is increased above a basal level of a housekeeping gene by at least approximately one point nine (1.9) standard deviations in the cancerous tumor; (b) messenger RNA and/or gene product (protein) of a MYC gene compared to a housekeeping gene in increased above a basal level of a housekeeping gene by at least approximately one point nine (1.9) standard deviations in the cancerous tumor; and/or (c) messenger RNA and/or gene product (protein) of one or more TCR genes is decreased below a basal level of a housekeeping gene by at least approximately one point nine (1.9) standard deviations.

Embodiment S5. The illudofulvene analog of Embodiment S4, where the one or more TCR genes are selected from the group consisting of one or more ERCC genes, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment S6. The illudofulvene analog of Embodiments S4 or S5, where the one or more PTGR genes are selected from the group consisting of PTGR1 and PTGR2.

Embodiment S7. The illudofulvene analog of Embodiments S4-S6, where the one or more housekeeping genes are selecting from the group consisting of beta actin (ACTB), transferrin receptor (TFRC) and glyceraldehyde-3-phosphate dehydrogenase (GAPD) and ubiquitin.

Embodiment S8. The illudofulvene analog of Embodiments S4-S7, where the gene expression was measured by means selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment S9. The illudofulvene analog of Embodiment S8, where the primers selected to hybridize with the one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, SEQ ID NO:900-902.

Embodiment S10. An illudofulvene analog for use in a method of treating a patient with cancer, wherein the method includes (a) determining from a patient test sample a first relative ratio of expression of messenger RNA and/or gene product (protein) of one or more PTGR genes compared with a basal level of a housekeeping gene or determining a second relative ratio of expression of messenger RNA and/or gene product (protein) of a MYC gene compared with a basal level of a housekeeping gene; and (b) determining a third relative ratio of expression of the TCR genes compared with a basal level of a housekeeping gene; wherein the patient is treated with the illudofulvene analog if the first relative ratio of expression is increased by at least approximately one point nine (1.9) standard deviations and the third relative ratio of expression is decreased by at least approximately one point nine (1.9) standard deviations or the second relative ratio of expression is increased by at least approximately one point nine (1.9) standard deviations and the third relative ratio of expression is decreased by at least approximately one point nine (1.9) standard deviations.

Embodiment S11. The illudofulvene analog for the use of Embodiment S10, where the one or more TCR genes are selected from the group consisting of one or more ERCC genes, a XPA gene, a GTF series gene, a UV gene, and a USP gene.

Embodiment S12. The illudofulvene analog for the use of Embodiment S10 or S11, where the one or more PTGR genes are selected from the group consisting of PTGR1 and PTGR2.

Embodiment S13. The illudofulvene analog for the use of Embodiment S10 to S12, where the ratio of gene expression is measured by means selected from the group consisting of (i) quantification of gene mRNA expression by probe analysis, (ii) microarray analysis, (iii) Real Time-Polymerase Chain Reaction (RT-PCR), (iv) Transcription Coupled Repair (TCR) Tumor Mutation Panel and (v) primary tumor cell sensitivity.

Embodiment S14. The illudofulvene analog for the use of Embodiment S10 to S13, where the primers selected to hybridize with the one or more nucleic acid molecules are selected from the group consisting of SEQ ID NO:251-261, SEQ ID NO:264-274, SEQ ID NO:307-308, SEQ ID NO:309-310, SEQ ID NO: 329-440, SEQ ID NO: 451-475, SEQ ID NO:682-696, SEQ ID NO: 715-727, SEQ ID NO:897-899, SEQ ID NO:900-902.

Embodiment S15. The illudofulvene analog for the use of Embodiment S10 to S14, where the effective amount is administered in a dosage between: a lower limit of approximately 0.1 mg of the illudofulvene per kilogram of body weight of the patient; and an upper limit of approximately 5 mg of the illudofulvene per kilogram of body weight of the patient.

Embodiment S16. The illudofulvene analog for the use of Embodiment S10 to S15, where the illudofulvene is selected from the group consisting of 002, 008, 012, 039, 040, 41, 42, 43, 44, 46, 75, 76, 77, 78, 79, 88, 90, 94, 103, 104, 108, 110, 112, 113, 114, 115, 116, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 130, 141, 142, 146, 149, 150, 160, 164, 165, 177, 178, 184. 189, 190, 196, 198, 199, 201, 202, 203, 207, 208, 209, 210, 217, 221, 223, 233, 234, 235, 240, 244, 254, 255, 259, 262, 263, 266, 267, 268, 269, 275, 276, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 304, 305, 306, 310, 314, 315, 317, 321, 323, 324, 332, 333, 334, 335, 336, 337, 338, 339, 340, 345, 346, 347, 348, 351, 352, 354, 356, 357, 368, 359, 361, 362, 363, 364, 365, 366, 367, 369, 370, 371, 372, 373, 374, 375, 377, 378, 379, 380, 381, 382, 383, 384, 389, 392, 393, and 394.

Embodiment S17. An illudofulvene analog for use in the treatment of a cancerous tumor expressing a PTGR gene, a MYC gene or one or more TCR genes, wherein said expression includes (a) contacting a sample from a patient comprising one or more nucleic acid molecules with one or more means for determining (i) the regulation of expression of one or more PTGR genes; (ii) the regulation of expression of at least one TCR gene; (iii) the regulation of expression of at least one housekeeping gene; (b) determining if a cancerous tumor is sensitive to an illudofulvene analog based on a first relative ratio of expression of the one or more PTGR genes compared with the at least one housekeeping gene, when the first relative ratio of expression is high, and a second relative ratio of expression of the at least one TCR gene compared with the at least one housekeeping gene, when the second relative ratio of expression is deficient; or

(c) contacting the sample comprising one or more nucleic acid molecules with one or more means for determining (iv) the regulation of expression of a MYC gene; (v) the regulation of expression of two or more TCR genes; (vi) the regulation of expression of one or more housekeeping genes; and (d) determining if the cancerous tumor is sensitive to the illudofulvene analog based on a third relative ratio of expression of the MYC gene compared with the one or more housekeeping genes, when the third relative ratio of expression is high, and a fourth relative ratio of expression of two or more TCR genes compared with the one or more housekeeping genes, when the fourth relative ratio of expression is deficient.

Embodiment S18. An illudofulvene analog for use in a method of treating a patient with cancer, wherein the method includes (a) determining from a patient test sample a first relative ratio of expression based on expression of the one or more PTGR genes compared with expression of at least one housekeeping gene; (b) determining from the patient test sample a second relative ratio of expression based on expression of at least one TCR genes compared with expression of at least one housekeeping gene; and (c) determining that the patient is sensitive, if the first relative ratio of expression is high and the second relative ratio of expression is deficient, then administer an effective amount of the illudofulvene; or (d) determining from a patient test sample a third relative ratio of expression based on expression of a MYC gene compared with at least one of the housekeeping genes; (e) determining from a patient test sample a fourth relative ratio of expression based on expression of two or more TCR genes compared with at least one of the housekeeping genes; and (f) determining that the patient is sensitive, if the third relative ratio of expression is high and the fourth relative ratio of expression is deficient, then administer an effective amount of the illudofulvene.

Embodiment S20. A method of treating a cancerous tumor in a patient including (a) determining gene expression of at least two genes compared with one or more housekeeping genes selected from the group consisting of PTGR1 gene, PTGR2 gene, MYC gene and one or more TCR genes and (b) treating with an effective amount of an illudofulvene analog if: (I) the expression of one of the one or more PTGR genes compared to a housekeeping gene in the cancerous tumor is upregulated and the expression of one of the one or more TCR genes compared with a housekeeping gene is downregulated, or (II) the expression of the MYC gene compared to a housekeeping gene in the cancerous tumor is upregulated and the expression of two of the one or more TCR genes compared with at least one housekeeping gene is downregulated.

Embodiment S21. The method of Embodiment 20, where the patient is treated with an effective amount of an illudofulvene analog when the gene expression of PTGR1 gene compared with at least one housekeeping gene is upregulated, the gene expression of PTGR2 gene compared with at least one housekeeping gene is upregulated and the gene expression of one TCR gene of the one or more TCR genes compared with at least one housekeeping gene is downregulated.

Embodiment S22. A method of treating a cancerous tumor in a patient including: (a) determining from a patient test sample a first relative ratio of expression of a MYC gene compared with a housekeeping gene, (b) determining a second relative ratio of expression of a first TCR gene of one or more TCR genes compared with at least one housekeeping gene; (c) determining a third relative ratio of expression of a second TCR gene of the one or more TCR genes compared with at least one housekeeping gene, and (d) treating with an effective amount of an illudofulvene analog if the first relative ratio of expression is upregulated, the second relative ratio of expression is downregulated, and the third relative ratio of expression is downregulated.

The foregoing description of embodiments of the methods, systems, and components of the present invention has been provided for the purposes of illustration and description. It is not intended to be exhaustive or to limit the invention to the precise forms disclosed. Many modifications and variations will be apparent to one of ordinary skill in the relevant arts. For example, steps performed in the embodiments of the invention disclosed can be performed in alternate orders, certain steps can be omitted, and additional steps can be added. The embodiments were chosen and described in order to best explain the principles of the invention and its practical application, thereby enabling others skilled in the art to understand the invention for various embodiments and with various modifications that are suited to the particular used contemplated. Other embodiments are possible and are covered by the invention. Such embodiments will be apparent to persons skilled in the relevant art(s) based on the teachings contained herein. The breadth and scope of the present invention should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.