he="65.11mm" id="EMI-C00024" img-content="table" img-format="tif" wi="64.77mm" />

(E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5- carboxamido)-8-((1-methylazetidin-3-yl)methyl)- 8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a- tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12- dicarboxamide (Compound 14)

15

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(E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5- carboxamido)-8-((1-methylpiperidin-4-yl)methyl)- 8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a- tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12- dicarboxamide (Compound 15)

16

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(E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5- carboxamido)-8-(morpholinomethyl)-8,9,16,19-tetrahydro- 7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 16)

17

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(E)-15-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1- (1-(2-fluoroethyl)-3-methyl-1H-pyrazole-5-carboxamido)- 8-morpholino-8,9,16,19-tetrahydro-7H-6,10-dioxa- 2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10- c′d′]diindene-4,12-dicarboxamide (Compound 17)

18

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(E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5- carboxamido)-8-((4-methylpiperazin-1-yl)methyl)- 8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a- tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12- dicarboxamide (Compound 18)

19

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(E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5- carboxamido)-8-(1H-imidazol-1-yl)-8,9,16,19-tetrahydro- 7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 19)

20

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(33R,35R,E)-12,62-bis(1-ethyl-3-methyl-1H-pyrazole-5- carboxamido)-11H,61H-2,5-dioxa-1,6(7,1)- dibenzo[d]imidazola-3(3,5)-pyrrolidinacyclodecaphan-8- ene-15,65-dicarboxamide (Compound 20)

21

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(33R,35R,E)-12,62-bis(1-ethyl-3-methyl-1H-pyrazole-5- carboxamido)-31-methyl-11H,61H-2,5-dioxa-1,6(7,1)- dibenzo[d]imidazola-3(3,5)-pyrrolidinacyclodecaphan-8- ene-15,65-dicarboxamide (Compound 21)

22

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(E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5- carboxamido)-8-(4-hydroxypiperidin-1-yl)-8,9,16,19- tetrahydro-7H-6,10-dioxa-2,14,15a,19a- tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12- dicarboxamide (Compound 22)

23

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(E)-8-(4-aminopiperidin-1-yl)-1,15-bis(1-ethyl-3-methyl- 1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H- 6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 23)

24

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(S,E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H- pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10- dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diinden-8-yl)pyrrolidin-3-yl dihydrogen phosphate (Compound 24)

25

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(E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H- pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10- dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diinden-8-yl)piperidin-4-yl dihydrogen phosphate (Compound 25)

26

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(E)-8-(3-cyanopyrrolidin-1-yl)-1,15-bis(1-ethyl-3-methyl- 1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H- 6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 26)

27

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(E)-8-(3-aminopyrrolidin-1-yl)-1,15-bis(1-ethyl-3-methyl- 1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H- 6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 27)

28

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(R,E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H- pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10- dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diinden-8-yl)pyrrolidin-3-yl dihydrogen phosphate (Compound 28)

29

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(E)-8-(4-cyanopiperidin-1-yl)-1,15-bis(1-ethyl-3-methyl- 1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H- 6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 29)

30

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(E)-8-(azetidin-3-yl)-1,15-bis(1-ethyl-3-methyl-1H- pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10- dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 30)

31

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(E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5- carboxamido)-8-(piperidin-4-yl)-8,9,16,19-tetrahydro-7H- 6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 31)

32

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(E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H- pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10- dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diinden-8-yl)pyrrolidine-3-carboxylic acid (Compound 32)

33

[Image Omitted]

(E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H- pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10- dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diinden-8-yl)piperidine-4-carboxylic acid (Compound 33)

34

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(E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5- carboxamido)-8-(1-methylazetidin-3-yl)-8,9,16,19- tetrahydro-7H-6,10-dioxa-2,14,15a,19a- tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12- dicarboxamide (Compound 34)

35

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(R,E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5- carboxamido)-8-(3-methoxypyrrolidin-1-yl)-8,9,16,19- tetrahydro-7H-6,10-dioxa-2,14,15a,19a- tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12- dicarboxamide (Compound 35)

36

[Image Omitted]

(E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5- carboxamido)-8-(pyridin-2-yl)-8,9,16,19-tetrahydro-7H- 6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1- cd:8,9,10-c′d′]diindene-4,12-dicarboxamidexamide (Compound 36)

In a further embodiment, the present invention provides a pharmaceutical composition comprising the compound of Formula (I), Formula (Ia), Formula (Ib) or pharmaceutically acceptable salts thereof and at least one or more pharmaceutically acceptable excipient.

In a further embodiment, the present invention provides a compound of Formula (I), Formula (Ia), Formula (Ib) or pharmaceutically acceptable salts thereof for use in the treatment of a disease or condition in which activation of STING is beneficial.

In a further embodiment, the present invention provides the use of a compound or pharmaceutical composition of Formula (I), Formula (Ia), Formula (Tb) or pharmaceutically acceptable salts thereof in the manufacture of a medicament for the treatment of a disease or condition in which activation of STING is beneficial.

In a further embodiment, the present invention provides a method of treatment of a disease or condition in which activation of STING is beneficial in a subject comprising administering a therapeutically effective amount of a compound of Formula (I), Formula (Ia), Formula (Ib) or its pharmaceutically acceptable salt thereof.

In a further embodiment, the invention provides a method of treatment of disease or condition selected from cancer and infectious diseases, in a mammal in need thereof, which comprises administering to such mammal a therapeutically effective amount of a compound of Formula (I), Formula (Ia), Formula (Ib) or its pharmaceutically acceptable salt thereof.

In a further embodiment, the invention provides a method of treatment of cancer such as solid tumors, leukemias and lymphomas.

In a further embodiment, the invention provides a method of treatment of infectious diseases such as viral infection or bacterial infection.

In a further embodiment, the invention provides a composition comprising compound of Formula (I), Formula (Ia), Formula (Tb) or pharmaceutically acceptable salts thereof, and one or more additional therapies.

In a further embodiment, the invention provides a vaccine adjuvant comprising a compound of Formula (I), Formula (Ia), Formula (Tb) or pharmaceutically acceptable salts thereof.

In a further embodiment, the invention provides a vaccine composition comprising compound of Formula (I), Formula (Ia), Formula (Tb) or pharmaceutically acceptable salts thereof, and an antigen or antigen composition.

DETAILED DESCRIPTION OF THE INVENTION

General terms used in formula can be defined as follows; however, the meaning stated should not be interpreted as limiting the scope of the term per se.

The term “alkyl” refers to an alkane derived hydrocarbon radical that includes solely carbon and hydrogen atoms in the backbone, contains no unsaturation, has from one to six carbon atoms, and is attached to the remainder of the molecule by a single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl) and the like. Unless set forth or recited to the contrary, all alkyl groups described or claimed herein may be straight chain or branched, substituted or unsubstituted.

The term ‘perhaloalkyl’, as used herein, means an alkyl group as defined hereinabove wherein all the hydrogen atoms of the said alkyl group are substituted with halogen. The perhaloalkyl group is exemplified by trifluoromethyl, pentafluoroethyl, and the like.

The term “aromatic ring” as used herein, refers to a monocyclic aromatic hydrocarbon ring system.

The term “heteroaromatic ring” as used herein, refers to a 5-6 membered monocyclic aromatic ring system having 1-2 ring heteroatoms selected from O, N, or S.

The term “cycloalkyl” or ‘carbocycle’ refers to a non-aromatic mono or multicyclic ring system having 3 to 12 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like. Unless set forth or recited to the contrary, all cycloalkyl groups described or claimed herein may be substituted or unsubstituted.

The term “C₃-C₅monocyclic cycloalkyl” refers to a substituted or unsubstituted non-aromatic monocyclic ring system having 3 to 5 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, and the like.

The term “aromatic carbocyclic ring”, as used herein, refers to aromatic hydrocarbon ring system. Examples include benzene ring, and the like.

The term ‘aryl’, as used herein, refers to a monovalent monocyclic, bicyclic or tricyclic aromatic hydrocarbon ring system. Examples of aryl groups include phenyl, naphthyl, anthracenyl, fluorenyl, indenyl, azulenyl, and the like.

The term ‘heteroaryl’, as used herein, refers to a 5-14 membered monocyclic, bicyclic, or tricyclic ring system having 1-4 ring heteroatoms selected from O, N, or S, and the remainder ring atoms being carbon (with appropriate hydrogen atoms unless otherwise indicated), wherein at least one ring in the ring system is aromatic.

The term “heterocycle” or “heterocyclyl”, unless otherwise specified, refers to substituted or unsubstituted non-aromatic, monocyclic, bicyclic, tricyclic or bridged/fused/spiro ring system having 3- to 15-membered ring which consists of carbon atoms and with one or more (e.g., 2 or 3) heteroatom(s) independently selected from N, O, S, P(O)(OR⁴), P(O)(R^4a) or S(O)₂. The point of attachment may be from any suitable carbon or nitrogen.

The term ‘oxo’ means a divalent oxygen (═O) attached to the parent group. For example oxo attached to carbon forms a carbonyl, oxo substituted on cyclohexane forms a cyclohexanone, and the like.

The term “tautomer” or “tautomeric form” refers to structural isomers of different energies which are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol isomerizations. Valence tautomers include interconversions by reorganization of some of the bonding electrons.

The term “geometric isomer” refers to E or Z geometric isomers (for example, cis or trans) of double bond.

All tautomeric forms and their possible geometrical isomers, including, but not limiting to Formula (s), Formula (r), and Formula (t), of the formulas and compounds described herein are intended to be encompassed within the scope of the present invention.

[Image Omitted]

The compounds of the present invention may have one or more chiral centers. The absolute stereochemistry at each chiral center may be ‘R’ or ‘S’. The compounds of the invention include all diastereomers and enantiomers and mixtures thereof. Unless specifically mentioned otherwise, reference to one stereoisomer applies to any of the possible stereoisomers. Whenever the stereoisomeric composition is unspecified, it is to be understood that all possible stereoisomers are included.

The term “stereoisomer” refers to a compound made up of the same atoms bonded by the same bonds but having different three-dimensional structures which are not interchangeable. The three-dimensional structures are called configurations. As used herein, the term “enantiomer” refers to two stereoisomers whose molecules are nonsuperimposable mirror images of one another. The term “chiral center” refers to a carbon atom to which four different groups are attached. As used herein, the term “diastereomers” refers to stereoisomers which are not enantiomers. The terms “racemate” or “racemic mixture” refer to a mixture of equal parts of enantiomers.

The term “treating” or “treatment” of a state, disease, disorder, condition or syndrome includes: (a) preventing or delaying the appearance of clinical symptoms of the state, disease, disorder, condition or syndrome developing in a subject that may be afflicted with or predisposed to the state, disease, disorder, condition or syndrome but does not yet experience or display clinical or subclinical symptoms of the state, disease, disorder, condition or syndrome; (b) inhibiting the state, disease, disorder, condition or syndrome, i.e., arresting or reducing the development of the disease or at least one clinical or subclinical symptom thereof; c) lessening the severity of a disease disorder or condition or at least one of its clinical or subclinical symptoms thereof; and/or (d) relieving the disease, i.e., causing regression of the state, disorder or condition or at least one of its clinical or subclinical symptoms.

The term “subject” includes mammals, preferably humans and other animals, such as domestic animals; e.g., household pets including cats and dogs.

A “therapeutically effective amount” refers to the amount of a compound that, when administered to a subject in need thereof, is sufficient to cause a desired effect. The “therapeutically effective amount” will vary depending on the compound, the disease and its severity, age, weight, physical condition and responsiveness of the subject to be treated.

Compounds disclosed herein and their tautomeric forms, stereoisomers, prodrugs may be prepared, for example, by techniques well known in the organic synthesis and familiar to a practitioner ordinarily skilled in art of this invention. In addition, the processes described herein may enable the synthesis of the compounds of the present invention. However, these may not be the only means by which the compounds described in the invention may be synthesized. Further, the various synthetic steps described herein may be performed in alternate sequences in order to furnish the desired compounds.

In a further aspect, of the present invention, there is provided a compound of Formula (I), Formula (Ia), and Formula (Ib), or a pharmaceutically acceptable salt thereof for use in the treatment of a disease or condition in which activation of STING is beneficial.

In a further aspect of the present invention, there is provided the use of a compound or pharmaceutical composition of Formula (I), Formula (Ia), and Formula (Ib), or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for use in the treatment of a disease or condition in which activation of STING is beneficial.

In a further aspect of the present invention, there is provided a method of the treatment of a disease or condition in which activation of STING is beneficial in a subject comprising administering a therapeutically effective amount of a compound of Formula (I), Formula (Ia), and Formula (Ib), or its pharmaceutically acceptable salt thereof.

In a further aspect the invention provides a method of treatment of disease or condition selected from cancer and infectious diseases, in a mammal in need thereof, which comprises administering to such mammal a therapeutically effective amount of a compound of Formula (I), Formula (Ia), and Formula (Ib), or its pharmaceutically acceptable salt thereof.

In a further aspect the invention provides a method of treatment cancer such as solid tumors, leukemias and lymphomas.

In a further aspect the invention provides a method of treatment of infectious diseases such as viral infection or bacterial infection. Examples of solid tumors which may be treated with the compounds of present invention include, but are not limited to, breast cancer, pancreatic cancer, lung cancer, colon cancer, coloretal cancer, brain cancer, renal cancer, testicular cancer, cancer of urethra, rectal cancer, cancer of fallopian tubes, penile cancer, vaginal cancer, stomach cancer, skin cancer, melanoma, liver cancer, gastrointestinal stromal tumors, urothelial cancer, thyroid cancer, parathyroid gland cancer, adrenal cancer, bone cancer, oral cancer, ovarian cancer, uterine cancer, head and neck sqamous cell carcinoma, endometrial cancer, gall bladder cancer, bladder cancer, orophyrangeal cancer, lymph node cancer, glioblastoma, astrocytoma, glioblastoma multiforme or sarcomas of soft tissue, fibrosarcoma, chondrosarcoma, hemangioma, teratoma, lipoma, myxoma, fibroma, rhabdomyoma, teratoma, cholangiocarcinoma, Ewing's sarcoma. Examples of leukemia, which may be treated with the compounds of present invention include, but are not limited to Lymphoblastic T cell leukemia, Chronic myelogenous leukemia, Acute lymphoblastic T cell leukemia, Acute myelobastic leukemia, Hairy-cell leukemia, Chronic neutrophilic leukemia, Mantle cell leukemia, Acute megakaryocytic leukemia, Multiple myeloma, Megakaryoblastic leukemia, Erythroleukemia, Plasmacytoma, Promyelocytic leukemia, Chronic myelomonocytic leukemia, Myelodysplastic syndrome, Myelofibrosis, Chronic myelogenous leukemia, Polycythemia vera, Thrombocythemia, Chronic lymphocytic leukemia, Prolymphocytic leukemia, Hairy cell leukemia, Waldenstrom's macroglobulinemia, Castleman's disease, Chronic neutrophilic leukemia, Immunoblastic large cell leukemia, Plasmacytoma, and Leukemias in any other parts of body. Examples of lymphoma, which may be treated with the compounds of present invention include, but are not limited to, Hodgkin's disease, non-Hodgkin's lymphoma, Follicular lymphoma, Mantle cell lymphoma, Burkitt's lymphoma, Lymphoblastic T-cell lymphoma, Marginal zone lymphoma, Cutaneous T cell lymphoma, CNS lymphoma, Small lymphocytic lymphoma, Lymphoplasmacytic lymphoma, Diffuse large B-cell lymphoma (DLBCL), Peripheral T-cell lymphoma, Anaplastic large cell lymphoma, Primary mediastinal lymphoma, Mycosis fungoides, Small non-cleaved cell lymphoma, Lymphoblastic lymphoma, Immunoblastic lymphoma, Primary effusion lymphoma and HIV associated (or AIDS related) lymphomas. Examples of viral infection which may be treated with the compounds of present invention include, but are not limited to, human immune deficiency virus (HIV), Human papillomavirus (HPV), hepatitis C virus (HCV), hepatitis B virus (HBV), Influenza (Orthomyxoviridae), Alphavirus, Rotavirus, Sendai, vaccinia, respiratory synctical virus, Lassa virus (Arenaviridae), Rabies virus (Rhabdoviridae), West nile virus, Dengue virus, Japanese encephalitis virus, and other Flaviviridae, RNA virus, DNA virus, virus belonging to the family of Alphaflexiviridae, Astroviridae, Alphatetraviridae, Alvernaviridae, Asfarviridae, Ampullaviridae, Adenoviridae, Ascoviridae, Betaflexiviridae, Bromoviridae, Barnaviridae, Bicaudaviridae. Baculoviridae Closteroviridae, Caliciviridae, Carmotetraviridae, Clavaviridae, Corticoviridae, Dicistroviridae, Endornaviridae, Filoviridae, Globuloviridae, Guttaviridae, Geminiviridae, Hytrosaviridae, Leviviridae, Luteoviridae, Lipothrixviridae, Mesoniviridae, Marnaviridae, Metaviridae, Malacoherpesviridae, Nodaviridae, Nyamiviridae, Nimaviridae, Nanoviridae, Piconaviridae, Partitiviridae, Picobirnaviridae, Paramyxoviridae, Poxviridae, Pandoraviridae, Polymaviridae, Phycodnaviridae, Papillomaviridae, Polydnaviruses, Polymaviridae, Permutotetraviridae, Potyviridae, Retroviridae, Siphoviridae, Sphaerolipoviridae, Virgaviridae, Togaviridae, Turriviridae, Tectiviridae. Examples of bacterial infection which may be treated with the compounds of present invention include, but are not limited to, infections caused by bacteria belonging to Brucella, Clostridium, Clostrodium, Campylobacter, Enterococcus, Fransicella, Listeria, Legionella, Mycobacteria, Pseudomonas, Salmonella, Staphylococcus, Yersinia genus. In a further aspect, the invention provides a composition comprising compound of Formula (I), Formula (Ia), and Formula (Ib), or its pharmaceutically acceptable salt thereof, and one or more additional therapies.

In a further aspect, the invention provides a composition comprising compound of Formula (I), Formula (Ia), and Formula (Ib), or a pharmaceutically acceptable salt thereof, and one or more additional therapies such as chemotherapy, immunotherapy or radiotherapy.

Chemotherapy comprises administering one or more additional chemotherapeutic agents that may be used in combination with the compounds of Formula (I), Formula (Ia), and Formula (Ib), or a pharmaceutically acceptable salt thereof. chemotherapeutic agents that may be used in combination includes topoisomerase II inhibitors, anti-tumor antibiotics, anti-metabolites, retinoids, antiviral agents, abiraterone acetate, altretamine, anhydrovinblastine, auristatin, bexarotene, bicalutamide, BMS 184476, 2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl)benzene sulfonamide, bleomycin, N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-proly-1-Lproline-tbutylamide, cachectin, cemadotin, chlorambucil, cyclophosphamide, 3′,4′-didehydro-4′deoxy-8′-norvin-caleukoblastine, docetaxol, doxetaxel, cyclophosphamide, carboplatin, carmustine, cisplatin, cryptophycin, cyclophosphamide, cytarabine, dacarbazine (DTIC), dactinomycin, daunorubicin, decitabine dolastatin, doxorubicin (adriamycin), etoposide, 5-fluorouracil, finasteride, flutamide, hydroxyurea and hydroxyureataxanes, ifosfamide, liarozole, lonidamine, lomustine (CCNU), MDV3100, mechlorethamine (nitrogen mustard), melphalan, mivobulin isethionate, rhizoxin, sertenef, streptozocin, mitomycin, methotrexate, taxanes, nilutamide, onapristone, paclitaxel, prednimustine, procarbazine, RPR109881, stramustine phosphate, tamoxifen, tasonermin, taxol, tretinoin, vinblastine, vincristine, vindesine sulfate, and vinflunine.

Immunotherapy comprises administering one or more additional immunostimulatory agents that may be used in combination with the compound of Formula (I), Formula (Ia), and Formula (Ib), or a pharmaceutically acceptable salt thereof. Immunostimulatory agents that may be used in combination herein includes vaccine adjuvants, such as Toll-like receptor agonists, T-cell checkpoint blockers, CTLA4, PD-1, PD-L1, TIM3, OX40, LAG3, B7-H3, GITR, 4-1BB, ICOS, CD40 and KIR antibody. Examples of CTLA-4 and PD-1 antagonists include, but are not limited to, ipilimumab, tremelimumab, nivolumab, pembrolizumab, CT-011, AMP-224, and MDX-1106.

In a further aspect the invention provides a vaccine adjuvant comprising a compound of Formula (I), Formula (Ia), and Formula (Ib), or a pharmaceutically acceptable salt thereof.

In a further aspect the invention provides a vaccine composition comprising compound of Formula (I), Formula (Ia), and Formula (Ib), or a pharmaceutically acceptable salt thereof, and an antigen or antigen composition.

Antigens and adjuvants that may be used in combination with the compound of Formula (I), Formula (Ia), and Formula (Ib), or a pharmaceutically acceptable salt thereof disclosed herein include B7 costimulatory molecule, interleukin-2, interferon-α, interferon-γ, GM-CSF, CTLA-4 antagonists, OX-40 agonist, CD40 agonist, sargramostim, levamisol, vaccinia virus, Bacille Calmette-Guerin (BCG), liposomes, alum, detoxified endotoxins, mineral oils, surface active substances such as lipolecithin, pluronic polyols, polyanions, peptides, and oil or hydrocarbon emulsions. Adjuvants, such as aluminum hydroxide or aluminum phosphate, can be added to increase the ability of the vaccine to trigger, enhance, or prolong an immune response. Additional materials, such as cytokines, chemokines, and bacterial nucleic acid sequences, like CpG, a toll-like receptor (TLR) 9 agonist as well as additional agonists for TLR 2, TLR 4, TLR 5, TLR 7, TLR 8, TLR9, including lipoprotein, LPS, monophosphoryllipid A, lipoteichoic acid, imiquimod, resiquimod, and in addition retinoic acid-inducible gene I (RIG-T) agonists such as poly I:C, used separately or in combination with the described compositions are also potential adjuvants.

The pharmaceutical compositions may be administered by a variety of means including non-parenterally, parenterally, by inhalation spray, topically, or rectally in formulations containing pharmaceutically acceptable carriers, adjuvants and vehicles. Intra-tumoral (directly into the tumor mass) or peri-tumoral (around the tumor mass) administration of the compounds of the present invention.

General Methods of Preparation

The compound of formula described herein may be prepared by techniques known in the art. In addition, the compound of formula described herein may be prepared by following the reaction sequence as depicted in Scheme-1 and Scheme-2. Further, in the following schemes, where specific bases, acids, reagents, solvents, coupling agents, etc., are mentioned, it is understood that other bases, acids, reagents, solvents, coupling agents etc., known in the art may also be used and are therefore included within the scope of the present invention. Variations in reaction conditions, for example, temperature and/or duration of the reaction, which may be used as known in the art, are also within the scope of the present invention. All the isomers of the compound of formula in described in these schemes, unless otherwise specified, are also encompassed within the scope of this invention.

The processes described herein may enable the synthesis of the compounds of the present invention. However, these may not be the only means by which the compounds described in the invention may be synthesized. Further, the various synthetic steps described herein may be performed in alternate sequences to afford the desired compounds.

[Image Omitted]

Scheme 1 shows a method of preparation of the compound of the formula (I-A) [wherein ring C, m, G₁, G₂, p and R²are as defined earlier for Formula 1] from the compound of formula (2).

The compound of the formula (2), is reacted with 1-(chloromethyl)-4-methoxybenzene in presence of base such as cesium carbonate, potassium carbonate, in solvents such as dimethyl formamide, acetonitrile, THF or acetone to obtain to obtain the compound of formula (3).

The compound of the formula (3) as obtained in the previous step is reacted with compounds of formula (10) in presence of base such as DIPEA in solvents like ethanol, methanol, THF or DMSO to obtain the compounds of formula (4).

The compounds of the formula (4) is reacted with sodium dithionite in solvents such as methanol, ethanol or THF to obtain the compounds of formula (5). Coupling of compounds of the formula (5) with compounds of formula (8), by using EDC and base such as triethyl amine or diisopropyl amine in solvents like dimethyl formamide or tetrahydrofuran to give compounds of formula (6).

The compounds of the formula (6) as obtained in the previous step was treated with acid like trifluoracetic acid or hydrochloric acid to deprotect the PMB group and so as to obtain the compounds of formula (7)

The compounds of the formula (7) as obtained is reacted with compounds of formula (9), where, L is halogen (Chloro/bromo) or -OMs or -OTs in presence of base such as sodium hydroxide, sodium hydride, potassium carbonate or caesium carbonate in solvents like tetrahydrofuran or dimethyl formamide at a temperature of about 30-80° C. or higher to afford the compounds of formula (I-A).

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Scheme 2 shows a method of preparation of the compound of the formula (I-A) [wherein ring C, m, G₁, G₂, p and R²as defined earlier for Formula 1] from the compound of formula (11).

The compound of formula (11) was prepared according to the procedure described in WO2017/100594. The compound of the formula (11), is reacted with compounds of formula (9-A) under Mitsunobu condition in presence of reagents like DIAD or DEAD and TPP in solvents such as dimethyl formamide, acetonitrile or THF to obtain the compounds of formula (12).

The compound of the formula (11) is alternatively can be reacted with compounds of formula (9), where, L is halogen (Chloro/bromo) or -OMs or -OTs in presence of base such as sodium hydride, potassium carbonate or caesium carbonate in solvents like tetrahydrofuran or dimethyl formamide at a temperature of about 30-80° C. or higher to afford the compounds of formula (12).

The compounds of the formula (12) as obtained in the previous step is reacted with compounds of formula (10) in presence of base such as DIPEA in solvents like ethanol, methanol, THF or DMSO to obtain the compounds of formula (13).

The compounds of the formula (13) is reacted with sodium dithionite in solvents such as methanol ethanol or THF to obtain the compounds of formula (14). Coupling of compounds of the formula (14) with compounds of formula (8), by using EDC and base such as triethyl amine or diisopropyl amine in solvents like dimethyl formamide or tetrahydrofuran to give compounds of formula (15).

The compounds of the formula (15) as obtained in the previous step was treated with base like lithium hydroxide or sodium hydroxide for hydrolysis to obtain the compounds of formula (16).

The compounds of the formula (16) is reacted with coupling reagent like TBTU or EDC and ammonium chloride in solvents like tetrahydrofuran, dimethyl formamide to afford the compounds of formula (I-A).

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Scheme 3 shows a method of preparation of the compound of the formula (I-A) [wherein ring C, m, G₁, G₂, p and R²as defined earlier for Formula 1] from the compound of formula (17).

The compound of the formula (17), is reacted with compounds of formula (21) under Mitsunobu condition in presence of reagents like DIAD or DEAD and TPP in solvents such as dimethyl formamide, acetonitrile or THF to obtain the compounds of formula (18).

The compounds of the formula (18) is reacted with HCl in dioxane in solvents such as chlorinated solvent or tehtrahydrofuran to deprotect the methoxy methyl (MOM) group which upon further treatment with trifluoromethanesulfonic anhydride in presence of base such as 2,6-lutidine to give the compounds of formula (19).

The compounds of the formula (19) is reacted with ring A in presence of base such as triethyl amine in solvents like tehtrahydrofuran or chlorinated solvent to afford the compounds of formula (20).

The compounds of the formula (20) can be converted to the compounds of formula (I-A) by the procedures analogously depicted in the Scheme 2 for the conversion of compounds of formula (12) to the compounds of formula (I-A).

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Scheme 4 shows a method of preparation of the compound of the formula (I-A) [wherein ring C, m, G₁, G₂, p and R²as defined earlier for Formula 1] from the compound of formula (3).

The compound of the formula (3) is reacted with compound of formula (28) in presence of base such as DIPEA in solvents like ethanol, methanol, THF or DMSO to obtain the compounds of formula (22).

The compounds of the formula (22) is reacted with sodium dithionite in solvents such as methanol, ethanol or THF to obtain the compounds of formula (23). Coupling of compounds of the formula (23) with compounds of formula (8), by using EDC and base such as triethyl amine or diisoproyl amine in solvents like dimethyl formamide or tetrahydrofuran to give compounds of formula (24), which on Boc deprotection using HCl in dioxane to give compounds of formula (25).

The compounds of the formula (25) as obtained in the previous step is reacted with compound of formula (3) in presence of base such as DIPEA in solvents like ethanol, methanol, THF or DMSO to obtain the compounds of formula (26).

The compounds of the formula (26) is reacted with sodium dithionite in solvents such as methanol, ethanol or THF to obtain the compounds of formula (27). Coupling of compounds of the formula (27) with compounds of formula (8), by using EDC and base such as triethyl amine or diisopropyl amine in solvents like dimethyl formamide or tetrahydrofuran to give compounds of formula (6).

The compounds of the formula (6) as obtained in the previous step was treated with acid like trifluoracetic acid or hydrochloric acid to deprotect the PMB group to obtain the compounds of formula (7)

The compounds of the formula (7) as obtained is reacted with compounds of formula (9), where, L is halogen (Chloro/bromo) or -OMs or -OTs in presence of base such as sodium hydroxide, sodium hydride, potassium carbonate or caesium carbonate in solvents like tetrahydrofuran, dimethyl formamide at a temperature of about 30-80° C. or higher to afford the compounds of formula (I-A).

[Image Omitted]

Scheme 5 shows a method of preparation of the compounds of the formula (I-D) [wherein ring C, m, G₂, p and R²as defined earlier for Formula 1] from the compounds of formula (I-B).

The compounds of the formula (I-B), is reacted with HCl in methanol to obtain the compounds of formula (I-C), which was treated with base like lithium hydroxide or sodium hydroxide for hydrolysis to obtain the compounds of formula (I-D).

ABBREVIATIONS

- DMF: dimethylformamide
- DMSO: dimethyl sulfoxide
- THF: tetrahydrofuran
- CDI: 1,1′-Carbonyldiimidazole
- EDC: 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide
- TBTU: 2-(1H-Benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate
- HATU: 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide
- hexafluorophosphate
- DIPEA: diisopropyl amine
- NMP: N-Methyl-2-pyrrolidone
- TEA: triethyl amine
- KI: potassium iodide
- DCM: dichloromethane
- MeOH: methanol
- MOM: methoxy methyl
- Ms: methanesulfonyl
- Ts: 4-methylbenzenesulfonyl
- PMB: 4-methoxy benzyl
- DEAD: diethyl azodicarboxylate
- DIAD: diisopropyl azodicarboxylate
- TPP: triphenyl phosphine

Some of the representative examples of the present invention were prepared by following one or more reaction schemes as described above.

The invention is further illustrated by the following examples which are provided merely to be exemplary of the invention and do not limit the scope of the invention. The examples set forth below demonstrate the synthetic procedures for the preparation of the relative compounds. Certain modifications and equivalents will be apparent to those skilled in the art and are intended to be included within the scope of the invention.

Nomenclature of the compounds of the invention is according to ChemDraw Professional 16.0. Structures of the intermediates as well as the final compounds were confirmed by spectral data.

EXPERIMENTAL

Synthesis of Intermediates:

Intermediate 1:

Synthesis of 4-chloro-3-hydroxy-5-nitrobenzamide

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Step 1: Synthesis of 4-chloro-3-methoxy-5-nitrobenzamide

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To a suspension of 4-chloro-3-methoxy-5-nitrobenzoic acid (Prepared by following procedure as reported in WO2017/100594, 20.0 g, 86 mmol) and N,N-dimethyl formamide (0.669 ml, 8.64 mmol) in dichloromethane (400 ml) was added oxalyl chloride (8.32 ml, 95 mmol) dropwise at 0° C. and the reaction mixture was stirred for 30 minutes at 40° C. Upon disappearance of starting material, reaction mixture was concentrated in vacuum and dissolved in 500 ml dichloromethane and cooled to 0° C. Ammonium hydroxide (135 ml, 864 mmol, 25% aqueous solution) was added dropwise and the reaction mixture was warmed to room temperature and stirred for 1 hr. Progress of the reaction was monitored by TLC. Upon completion, the reaction mass was concentrated under reduced pressure. The obtained solid was filtered, washed with water and dried under vacuum to afford the title compound (19.0 g, 95% yield).

LCMS (ESI): m/z 230.1 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 8.31 (s, 1H), 8.06 (d, J=1.8 Hz, 1H), 7.89 (d, J=1.9 Hz, 1H), 7.79 (s, 1H), 4.02 (s, 3H).

Step 2: Synthesis of 4-chloro-3-hydroxy-5-nitrobenzamide (Intermediate 1)

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4-chloro-3-methoxy-5-nitrobenzamide (Step 1, 18.5 g, 80 mmol) was co evaporated with toluene (4×10 ml) till complete dryness, and then was suspended in dichloromethane (250 ml) and stirred at room temperature. To the reaction was added 1M solution in dichloromethane of boron tribromide (481 ml, 481 mmol) in dropwise manner. The slurry was stirred for 60 hr at 40° C. under nitrogen. Reaction was monitored by 1H NMR. Upon completion, the volatiles were removed under reduced pressure till complete dryness. The reaction mass was cooled to −78° C. and wet ice was added slowly to the reaction mixture (till fuming stops). The reaction mixture was stirred at room temperature for 15 minutes. The precipitate was filtered and washed with water and dried thoroughly under vacuum to obtain the title compound as off-white solid (16.0 g, 92% yield).

LCMS (ESI): m/z 216.2 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 11.50 (s, 1H), 8.16 (s, 1H), 7.92 (d, J=2.0 Hz, 1H), 7.72 (d, J=2.0 Hz, 1H), 7.63 (s, 1H).

Intermediate 2:

Synthesis of (1R,5S)-3-(1,3-dichloropropan-2-yl)-8-oxa-3-azabicyclo[3.2.1]octane

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Step-1: Synthesis of Diethyl 2-((1R,5S)-8-oxa-3-azabicyclo[3.2.1]octan-3-yl)malonate

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To a stirred solution of diethyl 2-bromomalonate (5.0 g, 20.91 mmol), in chloroform (50 ml) was added 8-oxa-3-azabicyclo[3.2.1]octane hydrochloride (4.69 g, 31.4 mmol) and TEA (14.58 ml, 105 mmol) at rt and heated the reaction mixture at 60° C. for 12 h. Reaction was monitored by using TLC. The reaction mixture was concentrated to get crude product which was purified by combi flash using 15-20% ethyl acetate in hexanes to get the title compound (3.5 g, 61.7% yield).

LCMS (ESI): m/z 271.08 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 4.20 (d, J=2.6 Hz, 3H), 4.15 (m, 4H), 2.70 (m, 2H), 2.61 (m, 2H), 1.92-1.74 (m, 2H), 1.74-1.64 (m, 2H), 1.20 (t, J=7.1 Hz, 6H).

Step-2: Synthesis of 2-((1R,5S)-8-oxa-3-azabicyclo[3.2.1]octan-3-yl)propane-1,3-diol

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Diethyl 2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)malonate (Step 1 Product, 3.50 g, 12.90 mmol) in THF (20 ml) was added to lithium aluminium hydride (2.448 g, 64.5 mmol) solution in THF (20 ml) at 0° C. The resulting solution was stirred at 25° C. for 5 h. The mixture was quenched by adding ethyl acetate 25 ml and sat. aq. NaCl solution dropwise. The solid obtained was filtered through celite pad and the filtrate was concentrated to get crude product which was purified by column chromatography using 10% methanol in dichloromethane to afford title compound (1.5 g, 62.1% yield).

LCMS (ESI): m/z 188.14 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 4.16 (m, 2H), 3.44 (m, 4H), 2.69 (m, 2H), 2.44 (m, 2H), 2.36 (m, 1H), 1.86-1.77 (m, 2H), 1.72-1.61 (m, 2H).

Step-3: Synthesis of (1R,5S)-3-(1,3-dichloropropan-2-yl)-8-oxa-3-azabicyclo[3.2.1]octane (Intermediate 2)

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To a stirred solution of 2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)propane-1,3-diol (Step 2 Product, 0.50 g, 2.67 mmol) in THF (10 ml) was added thionyl chloride (0.780 ml, 10.68 mmol) dropwise at 0° C. and stirred the reaction mixture at 25° C. for 16 h. Reaction was monitored by LCMS.

Reaction mixture was concentrated to dryness and washed with diethyl ether to get salt. This salt was neutralized by adding 10% sodium bicarbonate and extracted with ethyl acetate. Ethyl acetate was concentrated to get free base which was purified by silica gel column chromatography using 5-10% ethyl acetate in hexanes to get the title compound (0.20 g, 33.4% yield).

LCMS (ESI): m/z 224.89 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 4.20 (m, 2H), 3.78 (m, 2H), 3.69 (m, 2H), 2.91 (q, J=6.3 Hz, 1H), 2.73 (dd, J=11.0, 2.0 Hz, 2H), 2.54 (d, J=1.9 Hz, 2H), 1.90-1.77 (m, 2H), 1.77-1.61 (m, 2H).

Intermediate 3:

Synthesis of (1R,5S)-3-(1,3-dichloropropan-2-yl)-8-oxa-3-azabicyclo[3.2.1]octane
Step-1: benzyl 1-(2-fluoroethyl)-3-methyl-1H-pyrazole-5-carboxylate

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In a round bottom flask, benzyl 3-methyl-1H-pyrazole-5-carboxylate (20.0 g, 92 mmol), 2-fluoroethan-1-ol (6.52 g, 102 mmol) and triphenylphospine (31.5 g, 120 mmol) were dissolved into tetrahydrofuran (200 ml) followed by the dropwise addition of diisopropyl azadicarboxylate (23.38 ml, 120 mmol) into the reaction mixture. Reaction mixture was stirred at 25° C. for 60 min. Reaction was monitored by using TLC. Upon completion, the reaction was quenched by addition of water (50 ml) and extracted with ethyl acetate (250 ml×2). Organic layers were combined and washed with brine, dried over sodium sulphate and concentrated under reduced pressure to get crude, which was purified by combi flash chromatography (20% ethyl acetate in hexane) to yield the title compound as white solid (18 g, 74.2% yield).

LCMS (ESI): m/z 262.96 (M+H)⁺

¹H NMR (400 MHz, Chloroform-d) δ 7.47-7.32 (m, 5H), 6.71 (s, 1H), 5.33 (s, 2H), 4.93-4.79 (m, 3H), 4.71 (t, J=5.0 Hz, 1H), 2.30 (s, 3H).

Step-2: 1-(2-fluoroethyl)-3-methyl-1H-pyrazole-5-carboxylic acid

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In a round bottom flask, benzyl 1-(2-fluoroethyl)-3-methyl-1H-pyrazole-5-carboxylate (step 1, 17.5 g, 66.7 mmol) was dissolved into tetrahydrofuran (50 ml) followed by the dropwise addition of lithium hydroxide (2.397 g, 100 mmol). Reaction mixture was stirred at 25° C. for 15 hrs. Reaction was monitored by using TLC. The reaction mixture was concentrated under reduced pressure and a sticky solid which was dissolved in water. Ether (100 ml) wash was given ti the aqueous solution and after separation, the aqueous layer PH was adjusted ~2 by 1N HCl. Solid was precipitated out, stirred for 15 min and filtered through Buchner funnel and washed with water. Obtained solid was dried under vacuum to get title compound as white solid (10 g, 87% yield).

LCMS (ESI): m/z 173.01 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 13.32 (s, 1H), 6.64 (s, 1H), 4.83-4.73 (m, 2H), 4.73-4.63 (m, 2H), 2.19 (s, 3H).

Step 3: Synthesis of 1-(2-fluoroethyl)-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (Intermediate 3)

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To a stirred solution of 1-(2-fluoroethyl)-3-methyl-1H-pyrazole-5-carboxylic acid (step 2,14.0 g, 81 mmol) in DCM (100 ml), were added DMF (6.30 μl, 0.081 mmol) and oxalyl chloride (7.83 ml, 89 mmol). During addition, the bubbling was observed. After 1 hr stirring the reaction mixture was concentrated under reduced pressure and the crude product (acid chloride) was proceeded further for the next reaction. In another RBF, potassium thiocyanate (7.90 g, 81 mmol) was dissolved in acetone (10 ml). The reaction mixture was cooled at 0-5° C. for 2 hrs. The above intermediate (acid chloride) was dissolved in acetone (10 ml) and added slowly to the reaction mixture. Upon completion, hexane (50 ml) was added to the reaction mixture and the reaction mixture was concentrated under vacuum. Repeated the same procedure for 3 times. After the concentration, hexane (50 ml) was added and filtered the solid compound. The resulting clear yellow filtrate was concentrated under vacuum. A crude product was purified by comb flash using ethyl acetate in hexane. The desired product was eluted by 0-5% of ethyl acetate, (10 gm, 58% yield).

¹H NMR (400 MHz, Chloroform-d) δ 6.79 (s, 1H), 4.86-4.73 (m, 2H), 4.71-4.67 (m, 2H), 2.31 (s, 3H).

Example 1: (E)-8-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 1)

[Image Omitted]

Step-1: Synthesis of 4-chloro-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide

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To a solution of 4-chloro-3-hydroxy-5-nitrobenzamide (Intermediate 1, 15.0 g, 69.3 mmol) in dimethylformaamide (75 ml) was added cesium carbonate (24.82 g, 76.0 mmol) and the mixture was stirred for 10 minutes at room temperature. 1-(chloromethyl)-4-methoxybenzene (11.39 g, 72.7 mmol) was added slowly at same temperature and the reaction mixture was stirred at 25° C. for 16 h. Reaction was monitored by TLC and LCMS. With vigorous stirring, water (300 mL) was added dropwise and the resulting solid was stirred for 15 minutes. The obtained solid was filtered and washed with water (200 ml). The compound was triturated with pentane to afford the title compound (14 g, 60.0% yield).

¹H NMR (400 MHz, DMSO-d₆) δ 8.30 (s, 1H), 8.04 (m, 2H), 7.80 (s, 1H), 7.48-7.40 (m, 2H), 7.03-6.95 (m, 2H), 5.29 (s, 2H), 3.77 (s, 3H).

Step-2: Synthesis of (E)-4,4′-(but-2-ene-1,4-diylbis(azanediyl))bis(3-((4-methoxybenzyl) oxy)-5-nitrobenzamide)

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To a suspension of 4-chloro-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide (Step 1 Product, 14.0 g, 41.6 mmol) in ethanol (100 ml) was added (E)-but-2-ene-1,4-diamine dihydrochloride (synthesized by methods known in the literature, 3.31 g, 20.79 mmol) and DIPEA (21.49 g, 166 mmol) and the mixture was allowed to stir at 100° C. for 48 hr in a sealed tube. Progress of the reaction was monitored by TLC and LCMS. Upon completion, the reaction mixture was concentrated under reduced pressure. The crude sticky solid was stirred with methanol (25 ml) for 15 minutes and the solid was filtered through Buchner funnel with bed wash of methanol (5 ml) to afford the title product (14.1 gm, 49.4% yield).

LCMS (ESI): m/z 687.28 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 8.20 (d, J=1.9 Hz, 3H), 8.03 (s, 3H), 7.77-7.65 (m, 4H), 7.38-7.29 (m, 4H), 6.94-6.84 (m, 4H), 5.40 (t, J=3.0 Hz, 2H), 5.02 (s, 4H), 3.99 (s, 4H), 3.73 (s, 6H).

Step-3: Synthesis of (E)-4,4′-(but-2-ene-1,4-diylbis(azanediyl))bis(3-amino-5-((4-methoxybenzyl)oxy)benzamide)

[Image Omitted]

To a stirred suspension of (E)-4,4′-(but-2-ene-1,4-diylbis(azanediyl))bis(3-((4-methoxybenzyl)oxy)-5-nitrobenzamide) (Step 2 Product, 14.0 g, 20.39 mmol) in tetrahydrofuran (35 ml) and methanol (75 ml) was added Sodium dithionite (35.5 g solution in 38 ml of water, 204 mmol) at 0° C. Ammonium hydroxide (79.0 ml, 510 mmol) was added at same temperature. Reaction mixture was warmed to 25° C. and stirred at 25° C. for 15 hr. Progress of reaction was monitored by LCMS. Organic solvents were removed from the reaction mixture under vacuum. The crude product was dissolved in 15% of methanol in methylene dichloride (250 ml). A solid compound was precipitated out, stirred for 15 min. The solid was filtered out and the filtrate organic layer was concentrated udder reduced pressure. The compound was co-evaporated with toluene (100 ml) twice to remove the traces water to obtain the title compound (6.2 gm, 48.5% yield).

LCMS (ESI): m/z 627.46 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 7.63 (s, 1H), 7.47-7.31 (m, 5H), 7.21 (m, 3H), 7.01-6.80 (m, 9H), 5.61 (t, J=6.7 Hz, 2H), 4.97 (d, J=9.6 Hz, 4H), 4.65 (s, 1H), 3.81-3.73 (m, 3H), 3.72 (s, 6H), 3.48 (d, J=3.5 Hz, 4H).

Step-4: Synthesis of (E)-1,1′-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide)

[Image Omitted]

(E)-4,4′-(but-2-ene-1,4-diylbis(azanediyl))bis(3-amino-5-((4-methoxybenzyl)oxy)benzamide) (Step 3 Product, 6.0 g, 9.57 mmol) was dissolved in 50 mL DMF and the solution cooled to 0° C. and a solution of 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (Synthesized by methods reported in the literature, 4.67 g, 23.93 mmol) in 5 mL DMF was added to it. The reaction mixture was stirred at 0° C. for 30 minutes after which 3-(ethyliminomethylideneamino)-N,N-dimethylpropan-1-amine; hydrochloride (5.51 g, 28.7 mmol) was added to the reaction mixture followed by the addition of diisopropylethylamine (10.02 mL, 57.4 mmol). The reaction mixture was allowed to warm to RT and stirred overnight. The progress of the reaction was monitored by LCMS and upon completion, the reaction mixture was concentrated under reduced pressure and water was added. The solid precipitate obtained was filtered to get the product (6.2 g, 68.2% yield).

LCMS (ESI): m/z 949.82 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.92 (s, 2H), 7.99 (s, 2H), 7.74 (s, 2H), 7.41 (m, 4H), 7.34-7.00 (m, 8H), 6.67 (d, J=8.4 Hz, 2H), 6.52 (s, 2H), 5.48 (s, 2H), 4.80 (m, 6H), 4.53 (d, J=7.5 Hz, 4H), 3.61 (s, 4H), 2.24-2.00 (m, 6H), 1.27 (m, 8H).

Step-5: Synthesis of (E)-1,1′-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazole-5-carboxamide)

[Image Omitted]

To a stirred solution of (E)-1,1′-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide) (Step 4 Product, 5.80 g, 6.11 mmol) in 50 mL dichloromethane was added trifluoroacetic acid (50 ml, 306 mmol) at room temperature. The reaction mixture was stirred for 1 h and after the completion of the reaction as judged from TLC, the volatiles were concentrated, and water was added to the residue resulting in the precipitation of the crude product. The solid was filtered and washed with water and dried by azeotropic distillation with toluene. The dried crude product was purified by trituration successively with diethyl ether, dichloromethane and acetonitrile and filtered to obtain the product (4.0 gm, 92% yield).

LCMS (ESI): m/z 709.32 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.77 (s, 2H), 10.45 (m, 2H), 7.82 (s, 2H), 7.44 (m, 3H), 7.18 (m, 3H), 6.50 (s, 2H), 5.95 (s, 2H), 4.95 (s, 4H), 4.51 (d, J=7.0 Hz, 4H), 2.09 (m, 6H), 1.24 (t, J=7.1 Hz, 6H).

Step 6: Synthesis of (E)-8-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 1)

[Image Omitted]

To a stirred solution of (E)-1,1′-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazole-5-carboxamide) (Step 5 Product, 200 mg, 0.282 mmol) in DMF (4 ml) was added 3-(1,3-dichloropropan-2-yl)-8-oxa-3-azabicyclo[3.2.1]octane (Intermediate 2, 95 mg, 0.423 mmol), sodium hydroxide (45.1 mg, 1.129 mmol) and potassium iodide (23.42 mg, 0.141 mmol). Then the reaction mixture was heated at 60° C. for 10 h. Reaction was monitored by LCMS. Reaction mixture was concentrated to dryness and washed with acetonitrile and purified by preparative HPLC to obtain the title compound (18 mg, 7.04% yield).

LCMS (ESI): m/z 860.68 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.88 (brs, 2H), 8.00 (brs, 2H), 7.69 (d, J=1.2 Hz, 2H), 7.58 (d, J=1.4 Hz, 2H), 7.41 (brs, 2H), 6.52 (s, 2H), 5.49 (s, 2H), 5.02-4.76 (m, 4H), 4.59 (dd, J=10.4, 4.2 Hz, 2H), 4.53 (q, J=7.1 Hz, 4H), 4.35-4.25 (m, 2H), 4.19 (dd, J=10.4, 6.4 Hz, 2H), 3.08 (s, 2H), 2.92-2.83 (m, 2H), 2.76-2.65 (m, 2H), 2.08-2.02 (m, 7H), 1.82-1.70 (m, 2H), 1.27 (t, J=7.1 Hz, 6H).

Example 2: (E)-8-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 2)

[Image Omitted]

Step 1: Synthesis of diethyl 5,5′-((2-(methoxymethoxy)propane-1,3-diyl)bis(oxy))bis(4-chloro-3-nitrobenzoate)

[Image Omitted]

To a cooled solution of ethyl 4-chloro-3-hydroxy-5-nitrobenzoate (32.5 g, 132.3 mmol), 2-(methoxymethoxy)propane-1,3-diol (9.0 g, 66.2 mmol), triphenylphosphine (69.4 g, 264.6 mmol)) in THF (200 ml) was added DIAD (51.5 ml, 264.6 mmol) in dropwise manner at room temperature and reaction mixture was stirred at 60° C. for 16 h. The progress of the reaction was monitored by TLC. Reaction mixture was concentrated to get crude material, to this IPA (200 ml) was added and stirred for 30 min at room temperature and then at −78° C. for 30 min, filtered solid, dried well to afford the title compound, (23.0 g 59% yield).

¹H NMR (400 MHz, Chloroform-d) δ 8.08 (d, J=1.7 Hz, 2H), 7.84 (d, J=1.8 Hz, 2H), 4.91 (s, 2H), 4.58-4.40 (m, 9H), 3.48 (s, 3H), 1.44 (t, J=7.1 Hz, 6H).

Step 2: Synthesis of diethyl 5,5′-((2-hydroxypropane-1,3-diyl)bis(oxy))bis(4-chloro-3-nitrobenzoate)

[Image Omitted]

To a cooled solution of diethyl 5,5′-((2-(methoxymethoxy)propane-1,3-diyl)bis(oxy))bis(4-chloro-3-nitrobenzoate) (Step 1, 23.0 g, 38.9 mmol) in DCM (150 ml) and Ethanol (150 ml) was added HCl in dioxane (100 ml, 388.9 mmol), and reaction mixture was stirred at 25° C. for 12 h. The progress of the reaction was monitored by TLC. Reaction mixture was concentrated and dissolved in DCM (50 ml) and washed by 8% sodium bicarbonate solution and by water, organic phase was dried over sodium sulphate and distilled under reduced pressure to get the title compound, (21.0 g, 97% yield).

¹H NMR (400 MHz, Chloroform-d) δ 8.10 (d, J=1.7 Hz, 2H), 7.85 (d, J=1.8 Hz, 2H), 4.61 (p, J=5.1 Hz, 1H), 4.50-4.40 (m, 6H), 3.72 (s, 3H), 1.44 (t, J=7.1 Hz, 6H).

Step 3: Synthesis of diethyl 5,5′-((2-(((trifluoromethyl)sulfonyl)oxy)propane-1,3-diyl)bis(oxy))bis(4-chloro-3-nitrobenzoate)

[Image Omitted]

To a cooled solution of diethyl 5,5′-((2-hydroxypropane-1,3-diyl)bis(oxy))bis(4-chloro-3-nitrobenzoate) (Step 2, 21.0 g, 38.4 mmol), in DCM (200 ml) 2,6-lutidine (17.9 ml, 153.5 mmol), trifluoromethanesulfonic anhydride (16.2 g, 57.6 mmol)) was added dropwise and reaction mixture was stirred at 0° C. for 1 h. The progress of the reaction was monitored by TLC.

Reaction mixture was quenched with 2N aqueous HCl and extracted by DCM and washed by water, dried over sodium sulphate and concentrated under reduced pressure to get the title compound, (24.0 g, 92% yield).

¹H NMR (400 MHz, Chloroform-d) δ 8.15 (d, J=1.7 Hz, 2H), 7.83 (d, J=1.7 Hz, 2H), 5.64 (p, J=4.8 Hz, 1H), 4.70-4.66 (m, 3H), 4.47 (q, J=7.1 Hz, 4H), 3.73 (s, 1H), 1.45 (t, J=7.1 Hz, 6H).

Step 4: Synthesis of diethyl 5,5′-((2-((1R,5S)-3-oxa-8-azabicyclo[3.2.1]octan-8-yl)propane-1,3-diyl)bis(oxy))bis(4-chloro-3-nitrobenzoate)

[Image Omitted]

To a cooled solution of (1R,5S)-3-oxa-8-azabicyclo[3.2.1]octane hydrochloride (10.1 g, 67.7 mmol) in THF (30 ml) was added, TEA (18.9 ml, 135.4 mmol) and stirred for 30 min at 0° C. and then added diethyl 5,5′-((2-(((trifluoromethyl)sulfonyl)oxy)propane-1,3-diyl)bis(oxy))bis(4-chloro-3-nitrobenzoate) (Step 3, 23.0 g, 33.9 mmol), stirred at 25° C. for 12 h. The progress of the reaction was monitored by TLC. Reaction mixture was concentrated and purified by column chromatography to get the title compound, (18.0 g, 83% yield).

LCMS (ESI): m/z 641.47 [M+1]

¹H NMR (400 MHz, Chloroform-d) δ 8.07 (d, J=1.7 Hz, 2H), 7.94-7.78 (m, 2H), 4.52-4.29 (m, 8H), 3.77 (d, J=10.5 Hz, 2H), 3.60 (d, J=10.1 Hz, 2H), 3.50-3.33 (m, 2H), 3.33-3.21 (m, 1H), 2.11-1.93 (m, 4H), 1.44 (t, J=7.1 Hz, 6H).

Step 5: Synthesis of diethyl (E)-7-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1,13-dinitro-7,8,14,15,18,19-hexahydro-6H-dibenzo[b,j][1,12]dioxa[4,9]diazacyclopentadecine-3,11-dicarboxylate

[Image Omitted]

To a solution of diethyl 5,5′-((2-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)propane-1,3-diyl)bis(oxy))bis(4-chloro-3-nitrobenzoate) (Step 4, 17.9 g, 27.9 mmol) in ethanol (300 ml) in 1 lit autoclave was added (E)-but-2-ene-1,4-diamine dihydrochloride (5.7 g, 36.2 mmol) and DIPEA (38.9 ml, 22.29 mmol) and stirred the reaction mixture at 110° C. for 24 h. The progress of the reaction was monitored by TLC. Reaction mixture was cooled to room temperature and the solid obtained was filtered and dried to get the title compound, (15.5 g, 85% yield).

LCMS (ESI): m/z 656.22 [M+1]

¹H NMR (400 MHz, Chloroform-d) δ 8.57 (d, J=1.9 Hz, 2H), 8.46 (m, 2H), 7.64 (d, J=1.9 Hz, 2H), 5.76 (s, 2H), 4.47 (dd, J=16.4, 6.8 Hz, 2H), 4.39 (q, J=7.1 Hz, 4H), 4.26 (dd, J=9.7, 5.1 Hz, 2H), 4.06 (dd, J=16.0, 7.0 Hz, 2H), 3.88-3.78 (m, 4H), 3.63 (d, J=10.5 Hz, 2H), 3.46-3.40 (m, 2H), 3.16 (t, J=5.3 Hz, 1H), 2.04 (s, 4H), 1.41 (t, J=7.1 Hz, 6H).

Step 6: Synthesis of diethyl (E)-1,13-diamino-7-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-7,8,14,15,18,19-hexahydro-6H-dibenzo[b,j][1,12]dioxa[4,9]diazacyclopentadecine-3,11-dicarboxylate

[Image Omitted]

To a stirred suspension of diethyl (E)-7-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1,13-dinitro-7,8,14,15,18,19-hexahydro-6H-dibenzo[b,j][1,12]dioxa[4,9]diazacyclopentadecine-3,11-dicarboxylate (Step 5, 15.0 g, 22.9 mmol) in THF (30 ml) and ethanol (30 ml) was added Sodium dithionite (39.8 g, 228.8 mmol) (solution in 5.0 ml of water) at 0° C. Ammonium hydroxide (71.3 ml, 457.6 mmol) was added at same temperature. Reaction mixture was warmed to 0-25° C. and stirred at 25° C. for 3 h. Progress of reaction was monitored by LCMS. Solid was filtered off and organic solvents were removed under vacuum. Water was added to the reaction mixture and extracted with DCM (4×50 ml). The organic layer was dried over sodium sulphate and concentrated to get crude residue. Solid was washed with ether to get the title compound, (13.0 g, 95% yield).

LCMS (ESI): m/z 596.08 [M+1]

Step 7: Synthesis of diethyl (E)-8-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxylate

[Image Omitted]

To a stirred solution of diethyl (E)-1,13-diamino-7-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-7,8,14,15,18,19-hexahydro-6H-dibenzo[b,j][1,12]dioxa[4,9]diazacyclopentadecine-3,11-dicarboxylate (Step 6, 14.0 g, 23.5 mmol) in DMF (200 ml), 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (8.3 g, 42.3 mmol) in 1 ml of DMF was added at 0° C. and stirred for 30 min followed by addition of EDC (18.0 g, 94.0 mmol) and TEA (26.2 ml, 188.0 mmol) were added at 0° C. and stirred for 18 hr. Reaction was monitored by LCMS. After completion, the mixture was concentrated under reduced pressure, water was added solid was filtered and dried to get crude material, which was purified by column chromatography (5% MeOH/DCM) to get the title compound, (14.0 g, 65% yield).

LCMS (ESI): m/z 918.07 [M+1]

¹H NMR (400 MHz, DMSO-d₆) δ 12.91 (s, 2H), 7.83 (d, J=1.3 Hz, 2H), 7.61 (d, J=1.4 Hz, 2H), 6.50 (s, 2H), 5.39 (s, 2H), 4.95 (d, J=16.0 Hz, 2H), 4.81 (d, J=16.4 Hz, 2H), 4.58 (dd, J=10.7, 4.4 Hz, 2H), 4.55-4.46 (m, 4H), 4.38-4.28 (m, 4H), 3.99-3.90 (m, 2H), 3.69 (d, J=10.1 Hz, 2H), 3.58 (d, J=4.2 Hz, 2H), 3.51 (d, J=10.2 Hz, 2H), 2.93 (t, J=5.1 Hz, 1H), 2.04 (s, 6H), 1.99-1.90 (m, 2H), 1.87-1.77 (m, 2H), 1.33 (t, J=7.1 Hz, 6H), 1.25 (t, J=7.1 Hz, 6H).

Step 8: Synthesis of (E)-8-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxylic acid

[Image Omitted]

To the stirred solution of diethyl (E)-8-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxylate (Step 7, 14.0 g, 15.3 mmol) in ethanol (70 ml) and THF (70 ml) was added lithium hydroxide hydrate (6.4 g, 152.5 mmol) by dissolving it in water (10 ml) at room temperature and stirred for 16 h at 50° C. The reaction was monitored by LCMS, after completion it was concentrated under reduced pressure, water was added and pH was adjusted to acidic by using 1N HCl. Solid was precipitate out, which was filtered and dried to get the title compound, (12.0 g, 91% yield).

LCMS (ESI): m/z 862.05 [M+1]

¹H NMR (400 MHz, DMSO-d₆) δ 7.94-7.81 (m, 2H), 7.70 (s, 2H), 5.55 (s, 2H), 5.20-4.97 (m, 4H), 4.86 (t, J=16.4 Hz, 2H), 4.64-4.39 (m, 8H), 3.81-3.63 (m, 5H), 2.41-2.32 (m, 2H), 2.15 (d, J=8.4 Hz, 2H), 2.06 (s, 6H), 1.26 (t, J=7.1 Hz, 6H).

Step 9: Synthesis of (E)-8-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 2)

[Image Omitted]

(E)-8-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxylic acid (Step 8, 10.0 g, 11.6 mmol) was dissolved in DMSO (80 ml). To this stirred solution were added HOBT (8.9 g, 58.0 mmol), EDC (11.1 g, 58.0 mmol) and DIPEA (16.2 ml, 92.8 mmol) in dropwise manner followed by addition of ammonium chloride (21.7 g, 406.1 mmol) at 0° C. Reaction mixture was allowed to stir at room temperature for overnight. The reaction was monitored by LCMS. After completion, the reaction mixture was poured into cold water, solid was precipitated out which was filtered & washed with water. Solid was dried well to get crude material. Crude product was purified by column chromatography, (DCM:MeOH) 20% and obtained solid was washed with acetonitrile and filtered to get the title compound, (6.0 g 60% yield).

LCMS (ESI): m/z 860.68 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.88 (s, 2H), 8.01 (s, 2H), 7.73 (s, 2H), 7.63 (s, 2H), 7.38 (s, 2H), 6.50 (s, 2H), 5.40 (s, 2H), 4.98 (d, J=16.4 Hz, 2H), 4.79 (d, J=16.5 Hz, 2H), 4.60 (dd, J=10.7, 4.6 Hz, 2H), 4.50 (q, J=7.2 Hz, 4H), 3.97 (dd, J=10.8, 5.5 Hz, 2H), 3.68 (d, J=10.1 Hz, 2H), 3.60 (s, 2H), 3.53 (d, J=10.0 Hz, 2H), 2.96-2.84 (m, 1H), 2.04 (s, 6H), 1.95 (s, 2H), 1.84-1.74 (m, 2H), 1.28-1.21 (m, 6H).

Example 3: (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-morpholino-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 3)

[Image Omitted]

The title compound was prepared by following the procedure outlined for Example 1 using (E)-1,1′-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazole-5-carboxamide) and 4-(1,3-dichloropropan-2-yl)morpholine and converting the product to the corresponding hydrochloride salt.

LCMS (ESI): m/z 834.67 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.24 (brs, 2H), 8.12 (brs, 2H), 7.95 (s, 2H), 7.81 (d, J=1.2 Hz, 2H), 7.51 (brs, 2H), 6.52 (s, 2H), 5.29 (s, 2H), 5.16 (d, J=9.0 Hz, 2H), 5.00-4.77 (m, 4H), 4.49 (q, J=7.1 Hz, 4H), 4.40 (dd, J=11.8, 6.0 Hz, 2H), 4.14-4.02 (m, 5H), 3.73-3.50 (m, 4H), 2.03 (s, 6H), 1.24 (t, J=7.1 Hz, 6H).

Example 4: (E)-8-(4,4-difluoropiperidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 4)

[Image Omitted]

The title compound was prepared by following the procedure outlined for Example 1 using (E)-1,1′-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazole-5-carboxamide) and 1-(1,3-dichloropropan-2-yl)-4,4-difluoropiperidine.

LCMS (ESI): m/z 868.67 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.86 (s, 2H), 8.32 (s, 1H), 8.01 (s, 2H), 7.67 (d, J=2.7 Hz, 4H), 7.43 (s, 2H), 6.55 (s, 2H), 5.58 (s, 2H), 4.90 (s, 4H), 4.66-4.46 (m, 7H), 4.26 (dd, J=10.6, 6.5 Hz, 2H), 2.94 (d, J=5.5 Hz, 4H), 2.08 (m, 10H), 1.27 (m, 6H).

Example 5: (E)-8-((2S,6R)-2,6-dimethylmorpholino)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 5)

[Image Omitted]

The title compound was prepared by following the procedure outlined for Example 1 using (E)-1,1′-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazole-5-carboxamide) and (2S,6R)-4-(1,3-dichloropropan-2-yl)-2,6-dimethylmorpholine and converting the product to the corresponding hydrochloride salt.

LCMS (ESI): m/z 862.80 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 8.14 (s, 2H), 7.97 (s, 2H), 7.80 (d, J=1.1 Hz, 2H), 7.49 (s, 2H), 6.52 (s, 2H), 4.92 (s, 3H), 4.50 (t, J=7.0 Hz, 4H), 4.39-4.28 (m, 4H), 3.80-3.68 (m, 4H), 3.38 (qd, J=7.1, 5.5 Hz, 6H), 3.17 (s, 3H), 2.03 (s, 6H), 1.22 (t, J=7.0 Hz, 6H), 1.09 (d, J=7.0 Hz, 6H).

Example 6: (S,E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(3-methoxypyrrolidin-1-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 6)

[Image Omitted]

The title compound was prepared by following the procedure outlined for Example 1 using (E)-1,1′-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazole-5-carboxamide) and (S)-1-(1,3-dichloropropan-2-yl)-3-methoxypyrrolidine.

LCMS (ESI): m/z 848.18 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.86 (s, 2H), 8.02 (s, 2H), 7.73 (s, 2H), 7.64 (s, 2H), 7.39 (s, 2H), 6.52 (s, 2H), 5.44 (s, 2H), 4.98-4.79 (m, 4H), 4.64 (d, J=10.6 Hz, 3H), 4.51 (d, J=7.8 Hz, 4H), 4.13 (s, 2H), 3.95 (s, 1H), 3.23 (s, 3H), 3.07 (d, J=11.4 Hz, 2H), 2.85 (m, 2H), 2.05 (s, 6H), 1.76 (s, 2H), 1.28 (t, J=7.1 Hz, 6H).

Example 7: (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(piperidin-1-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 7)

[Image Omitted]

LCMS (ESI): m/z 832.52 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.89 (s, 2H), 8.00 (s, 2H), 7.70 (s, 2H), 7.62 (d, J=5.2 Hz, 2H), 7.39 (s, 2H), 6.54 (s, 2H), 5.54 (s, 2H), 4.90 (m, 4H), 4.62 (dd, J=10.5, 4.3 Hz, 2H), 4.53 (q, J=7.4 Hz, 3H), 4.20 (dd, J=10.5, 6.4 Hz, 2H), 2.78 (t, J=5.2 Hz, 4H), 2.07 (s, 6H), 1.60 (s, 4H), 1.47 (s, 2H), 1.27 (t, J=6.8 Hz, 6H).

Example 8: (E)-8-(azetidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 8)

[Image Omitted]

The title compound was prepared by following the procedure outlined for Example 1 using (E)-1,1′-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazole-5-carboxamide) and 1-(1,3-dichloropropan-2-yl)azetidine hydrochloride.

LCMS (ESI): m/z 804.67 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.86 (brs, 2H), 7.98 (d, J=1.6 Hz, 2H), 7.66 (d, J=1.8 Hz, 2H), 7.35 (m, 4H), 6.45 (m, 2H), 6.12 (m, 2H), 5.12 (m, 1H), 5.05-4.88 (m, 3H), 4.65-4.34 (m, 6H), 4.28 (t, J=7.8 Hz, 1H), 3.65 (t, J=9.9 Hz, 1H), 2.08 (m, 2H), 1.89 (d, J=8.0 Hz, 6H), 1.80-1.69 (m, 1H), 1.55 (d, J=6.7 Hz, 1H), 1.34-1.18 (m, 6H).

Example 9: (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(1-methylpiperidin-4-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 9)

[Image Omitted]

The title compound as its formate salt was prepared by following the procedure outlined for Example 1 using (E)-1,1′-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazole-5-carboxamide) and 4-(1,3-dichloropropan-2-yl)-1-methylpiperidine.

LCMS (ESI): m/z 846.68 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.95 (s, 3H), 8.48 (s, 1H), 8.60-8.42 (m, 2H), 8.02-7.95 (m, 1H), 7.81 (s, 1H), 7.66 (s, 1H), 7.40 (d, J=7.4 Hz, 3H), 7.33 (s, 1H), 7.25 (s, 1H), 6.56 (s, 1H), 6.44 (s, 1H), 5.82 (m, 2H), 4.92 (m, 4H), 4.52 (m, 4H), 4.05 (d, J=7.6 Hz, 2H), 2.89 (s, 4H), 2.55-2.54 (s, 3H), 2.10 (d, J=21.4 Hz, 6H), 2.00 (s, 2H), 1.84 (d, J=7.5 Hz, 1H), 1.59 (s, 1H), 1.38-1.15 (m, 8H).

Example 10: (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(piperazin-1-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 10)

[Image Omitted]

The title compound as its dihydrochloride salt was prepared by following the procedure outlined for Example 1 using (E)-1,1′-(but-2-ene-1,4-diyl)bis(2-(I-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazole-5-carboxamide) and tert-butyl 4-(1,3-dichloropropan-2-yl)piperazine-1-carboxylate followed by Boc deprotection.

LCMS (ESI): m/z 833.80 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 8.07 (s, 2H), 7.79 (d, J=18.4 Hz, 4H), 7.45 (s, 2H), 6.53 (s, 2H), 5.43 (s, 1H), 4.91 (s, 6H), 4.51 (d, J=7.4 Hz, 5H), 3.46-3.30 (m, 9H), 2.04 (s, 6H), 1.27 (d, J=7.0 Hz, 6H).

Example 11: (S,E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(3-hydroxypyrrolidin-1-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 11)

[Image Omitted]

Step-1: Synthesis of diethyl (S)-2-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)malonate

[Image Omitted]

Diethyl (S)-2-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)malonate synthesized from diethyl 2-bromomalonate and (S)-3-((tert-butyldimethylsilyl)oxy)pyrrolidine hydrochloride by using the procedure disclosed in Step-1 of Intermediate 2.

LCMS (ESI): m/z 360.41 (M+H)⁺

¹H NMR (400 MHz, CDCl₃) δ 4.49-4.38 (m, 1H), 4.26 (q, J=7.0 Hz, 4H), 3.26 (dd, J=9.4, 6.3 Hz, 1H), 3.03-2.82 (m, 2H), 2.63 (dd, J=9.4, 4.8 Hz, 1H), 2.19-2.01 (m, 1H), 1.81-1.65 (m, 2H), 1.31 (t, 6H), 0.89 (s, 9H), 0.06 (s, 6H).

Step-2: Synthesis of (S)-2-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)propane-1,3-diol

[Image Omitted]

(S)-2-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)propane-1,3-diol synthesized from diethyl (S)-2-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)malonate (Step 1 product) by using the procedure disclosed in Step-2 of Intermediate 2.

LCMS (ESI): m/z 276.21 (M+H)⁺

¹H NMR (400 MHz, CDCl₃) δ 4.42 (m, 1H), 3.94-3.82 (m, 1H), 3.81-3.73 (m, 4H), 3.19 (dd, J=10.6, 5.3 Hz, 1H), 3.01-2.95 (m, 1H), 2.85-2.73 (m, 2H), 2.14-2.01 (m, 1H), 1.91-1.74 (m, 1H), 0.91 (s, 9H), 0.10 (s, 6H).

Step 3: Synthesis of dimethyl 5,5′-((2-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)propane-1,3-diyl)bis(oxy))(S)-bis(4-chloro-3-nitrobenzoate)

[Image Omitted]

To the stirred solution of methyl 4-chloro-3-hydroxy-5-nitrobenzoate (Synthesized by methods reported in the literature, 3.87 g, 16.70 mmol), (S)-2-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)propane-1,3-diol (Step-2 product, 2.3 g, 8.35 mmol) and triphenylphosphine (8.76 g, 33.4 mmol) in THF (10 ml) was added DIAD (6.49 ml, 33.4 mmol) in dropwise manner at 0-5° C. After the addition reaction mixture was stirred at 50° C. for 8 hr. After the completion of reaction as it was monitored by TLC, the volatiles were concentrated under reduced pressure and the crude purified by column chromatography to afford the title product as an off-white solid (2.3 g, 40%).

LCMS (ESI): m/z 702.58 (M+H)⁺

¹H NMR (400 MHz, CDCl₃) δ 8.13-8.03 (m, 2H), 7.89-7.76 (m, 2H), 4.48 (m, 5H), 3.99 (s, 6H), 3.67-3.43 (m, 1H), 3.16 (m, 3H), 2.79 (s, 1H), 2.21-1.89 (m, 2H), 0.89 (s, 9H), 0.08 (s, 6H).

Step 4: Synthesis of dimethyl (S,E)-7-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)-1,13-dinitro-7,8,14,15,18,19-hexahydro-6H-dibenzo[b,j][1,12]dioxa[4,9]diazacyclopentadecine-3,11-dicarboxylate

[Image Omitted]

To the stirred solution of dimethyl 5,5′-((2-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)propane-1,3-diyl)bis(oxy))(S)-bis(4-chloro-3-nitrobenzoate) (Step 3, 1.2 g, 1.71 mmol) in 30 ml ethanol were added (E)-but-2-ene-1,4-diamine dihydrochloride (0.27 g, 1.71 mmol) and DIPEA (1.79 ml, 10.25 mmol). The reaction mixture was stirred at 120° C. for 16 hr in a sealed tube. The progress of the reaction was monitored by LCMS and TLC. After the completion, the reaction mass was concentrated under reduced pressure and the crude purified by column chromatography to afford the title product as a red coloured solid. (0.76 g, 62%).

LCMS (ESI): m/z 716.77 (M+H)⁺

¹H NMR (400 MHz, Chloroform-d) δ 8.56 (d, J=2.1 Hz, 2H), 8.45 (t, J=6.6 Hz, 2H), 7.62 (d, J=2.0 Hz, 2H), 5.79 (d, J=1.5 Hz, 2H), 4.56-4.44 (m, 3H), 4.37 (q, J=7.1 Hz, 3H), 4.11-4.01 (m, 3H), 3.91 (s, 6H), 3.89-3.85 (m, 2H), 1.81-1.75 (m, 2H), 1.45-1.38 (m, 3H), 0.92 (s, 9H), 0.11 (d, J=1.3 Hz, 6H).

Step 5: Synthesis of dimethyl (S,E)-1,13-diamino-7-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)-7,8,14,15,18,19-hexahydro-6H-dibenzo[b,j][1,12]dioxa[4,9]diazacyclopentadecine-3,11-dicarboxylate

[Image Omitted]

To the solution of dimethyl (S,E)-7-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)-1,13-dinitro-7,8,14,15,18,19-hexahydro-6H-dibenzo[b,j][1,12]dioxa[4,9]diazacyclopentadecine-3,11-dicarboxylate (Step 4, 0.750 g, 1.048 mmol) in methanol:THF (15 ml:15 ml) was added solution of sodium dithionite (2.159 g, 10.48 mmol) in 10 ml water followed by the addition of ammonium hydroxide (4.08 ml, 26.2 mmol) at 0° C. Reaction was monitored by TLC and LCMS. Reaction mass was added to water (20 ml) and was extracted with ethyl acetate and dried over sodium sulphate and concentrated under reduced pressure to get crude which was purified by column chromatography to afford the title product (0.6 g, 87%).

LCMS (ESI): m/z 656.70 (M+H)⁺

Step 6: Synthesis of Dimethyl (S,E)-8-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxylate

[Image Omitted]

To a stirred solution of dimethyl (S,E)-1,13-diamino-7-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)-7,8,14,15,18,19-hexahydro-6H-dibenzo[b,j][1,12]dioxa[4,9]diazacyclopentadecine-3,11-dicarboxylate (Step 5, 0.65 g, 0.99 mmol) in 20 ml DMF was added 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (0.35 g, 1.78 mmol) at 0° C. The reaction mixture was stirred for 30 min followed by the addition of EDC (0.57 g, 2.97 mmol) and Et₃N (0.69 ml, 4.96 mmol) and the reaction mixture allowed to warm at room temperature for overnight. After the completion of the reaction, the mixture was concentrated under reduced pressure and water was added resulting in the precipitation of the product as a white solid which was filtered and dried. (800 mg, 83%).

LCMS (ESI): m/z 977.58 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.94 (brs, 2H), 7.83 (d, J=5.8 Hz, 2H), 7.62 (d, J=4.9 Hz, 2H), 6.53 (s, 2H), 5.49 (d, J=10.3 Hz, 2H), 4.92 (m, 4H), 4.71-4.47 (m, 6H), 4.43 (s, 2H), 4.33 (q, J=7.0 Hz, 2H), 4.21 (m, 2H), 3.87 (s, 6H), 3.13 (m, 1H), 2.99 (s, 1H), 2.88 (d, J=10.6 Hz, 1H), 2.78-2.65 (m, 2H), 2.05 (s, 6H), 1.66 (s, 1H), 1.26 (t, J=6.9 Hz, 6H), 0.88 (s, 9H), 0.08 (d, J=1.8 Hz, 6H).

Step 7: Synthesis of (S,E)-8-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxylic acid

[Image Omitted]

To a solution of Dimethyl (S,E)-8-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxylate (Step 6, 800 mg, 0.82 mmol) in 15 ml of a mixture of THF, methanol and water (1:1:1) was added lithium hydroxide (19.58 mg, 0.82 mmol) & stirred for 12 hr at room temperature. Reaction was monitored by LCMS. After the completion of reaction, the mixture was concentrated under reduced pressure to get a residue which was diluted with water. A solution of 1 N HCl was added until the pH 5 and the resulting white precipitate was filtered and dried to afford the title compound (640 mg, 84% yield).

LCMS (ESI): m/z 950.82 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.96 (brs, 4H), 7.86 (d, J=8.2 Hz, 2H), 7.83-7.62 (m, 2H), 6.54 (s, 2H), 5.47 (s, 2H), 5.12-4.76 (m, 8H), 4.51 (d, J=7.6 Hz, 8H), 4.30-3.98 (m, 1H), 2.80-2.63 (m, 2H), 2.17 (d, J=2.6 Hz, 1H), 2.05 (s, 6H), 1.29-1.25 (m, 6H), 0.87 (s, 9H), 0.10 (s, 6H).

Step 8: Synthesis of (S,E)-8-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide

[Image Omitted]

(S,E)-8-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxylic acid (Step 7, 650 mg, 0.684 mmol) was dissolved in 50 ml DMF. Ammonium chloride (1.1 g, 20.52 mmol), TBTU (661 mg, 2.052 mmol) followed by DIPEA (0.717 ml, 4.10 mmol) was added and the reaction mixture was allowed to stir at room temperature overnight. The reaction was monitored by LCMS, and after the completion of reaction, the mixture was concentrated and poured into crushed ice. The diamide as a white solid was obtained which was filtered and dried (600 mg, 92% yield).

LCMS (ESI): m/z 949.07 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.95 (brs, 2H), 7.78 (brs, 2H), 7.32 (brs, 2H), 7.19 (s, 2H), 7.06 (s, 2H), 6.52 (s, 2H), 5.34 (d, J=16.4 Hz, 2H), 5.11-4.76 (m, 8H), 4.50 (d, J=7.2 Hz, 6H), 3.71-3.50 (m, 2H), 3.13 (dd, J=7.3, 4.2 Hz, 2H), 2.16 (s, 2H), 2.04 (s, 6H), 1.30-1.20 (m, 6H), 0.87 (s, 9H), 0.10 (s, 6H).

Step 9: Synthesis of (S,E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(3-hydroxypyrrolidin-1-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide hydrochloride salt (Compound 11)

[Image Omitted]

(S,E)-8-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Step 8, 600 mg, 0.633 mmol) was stirred in 3 M HCl in methanol for 16 hrs at room temperature. The progress of the reaction was monitored by LCMS. After completion of the reaction, the volatiles were removed under reduced pressure to get the crude, which was triturated with ether to obtain the title compound which was further purified by preparative HPLC. (150 mg, 28% yield).

LCMS (ESI): m/z 834.80 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 8.19-8.00 (m, 2H), 7.89-7.76 (m, 4H), 7.48 (s, 2H), 6.54 (d, J=2.0 Hz, 2H), 5.34 (d, J=3.9 Hz, 2H), 4.94 (m, 6H), 4.50 (m, 10H), 4.06-3.37 (m, 2H), 2.55 (d, J=2.3 Hz, 2H), 2.04 (m, 6H), 1.26-1.23 (m, 6H).

Example 12: (R,E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(3-hydroxypyrrolidin-1-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 12)

[Image Omitted]

The title compound as its hydrochloride salt was prepared by following the procedure outlined for Example 11 using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and (R)-2-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)propane-1,3-diol

LCMS (ESI): m/z 834.80 (M+H)⁺

Example 13: (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(2-morpholinoethyl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 13)

[Image Omitted]

The title compound as its hydrochloride salt was prepared by following the procedure outlined for Example 1 using (E)-1,1′-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazole-5-carboxamide) and 4-(4-chloro-3-(chloromethyl)butyl)morpholine.

LCMS (ESI): m/z 862.51 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.90 (brs, 2H), 10.57 (brs, 1H), 7.98 (brs, 1H), 7.88 (brs, 1H), 7.67 (s, 1H), 7.44 (brs, 1H), 7.38 (s, 1H), 7.33 (s, 1H), 7.29 (brs, 1H), 7.19 (s, 1H), 6.55 (s, 1H), 6.48 (s, 1H), 5.87-5.74 (m, 2H), 5.05-4.80 (m, 4H), 4.63-4.42 (m, 4H), 4.05 (d, J=5.4 Hz, 4H), 3.87-3.31 (m, 6H), 2.68-2.60 (m, 4H), 2.35-2.25 (m, 3H), 2.13 (s, 3H), 2.09 (s, 3H), 1.29 (t, J=7.1 Hz, 6H).

Example 14: (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-((1-methylazetidin-3-yl)methyl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 14)

[Image Omitted]

LCMS (ESI): m/z 832.52 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.83 (s, 3H), 8.27 (s, 1H), 8.16-7.91 (m, 1H), 7.73 (s, 1H), 7.72-7.63 (m, 1H), 7.61 (s, 1H), 7.49-7.25 (m, 4H), 6.53 (s, 2H), 5.39 (s, 2H), 5.05-4.81 (m, 4H), 4.65-4.44 (m, 4H), 4.35 (m, 2H), 4.14 (dd, J=9.7, 6.7 Hz, 2H), 3.57 (t, J=7.3 Hz, 2H), 2.99 (s, 2H), 2.55 (s, 3H), 2.47-2.39 (m, 2H), 2.25-1.98 (m, 8H), 1.32-1.14 (m, 6H).

Example 15: (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-((1-methylpiperidin-4-yl)methyl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 15)

[Image Omitted]

LCMS (ESI): m/z 860.53 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.92 (brs, 2H), 8.31 (s, 1H), 8.02 (brs, 2H), 7.73 (d, J=1.2 Hz, 2H), 7.59 (s, 2H), 7.40 (brs, 2H), 6.53 (s, 2H), 5.39 (s, 2H), 4.98-4.79 (m, 4H), 4.51 (q, J=7.1 Hz, 4H), 4.37 (m, 2H), 4.16 (m, 2H), 2.79 (m, 2H), 2.46 (m, 2H), 2.17 (s, 3H), 2.06 (s, 6H), 1.94 (m, 2H), 1.75 (d, J=12.4 Hz, 2H), 1.50 (t, J=7.0 Hz, 2H), 1.25-1.23 (m, 8H).

Example 16: (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(morpholinomethyl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 16)

[Image Omitted]

LCMS (ESI): m/z 848.18 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.91 (brs, 2H), 8.24 (s, 1H), 8.01 (brs, 2H), 7.70 (d, J=1.2 Hz, 2H), 7.62 (s, 2H), 7.39 (brs, 2H), 6.53 (s, 2H), 5.39 (s, 2H), 4.88 (s, 4H), 4.61-4.38 (m, 6H), 4.23 (t, J=4 Hz, 2H), 3.68 (t, J=4.5 Hz, 4H), 2.06 (s, 8H), 1.28-1.22 (m, 6H).

Example 17: (E)-15-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1-(1-(2-fluoroethyl)-3-methyl-1H-pyrazole-5-carboxamido)-8-morpholino-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 17)

[Image Omitted]

Step-1: Tert-butyl (E)-(4-((4-carbamoyl-2-((4-methoxybenzyl)oxy)-6-nitrophenyl)amino) but-2-en-1-yl)carbamate

[Image Omitted]

In a 250 ml sealed tube, to a solution of 4-chloro-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide (Step 1 product of example 1, 24.00 g, 71.3 mmol) in ethanol (100 ml), tert-butyl (E)-(4-aminobut-2-en-1-yl)carbamate (19.91 g, 107 mmol) was added followed by addition of diisopropylethylamine (62.2 ml, 356 mmol) and heated at 120° C. for 16 hr. LCMS of the aliquot showed starting amide unreacted, so added another 0.6 eq of tert-butyl (E)-(4-aminobut-2-en-1-yl)carbamate and reaction continued for another 16 hr. Progress of the reaction was monitored by TLC. The mixture was cooled, the red solid was filtered and washed with ethanol. The compound was purified by combiflash chromatography using methanol in DCM. The desired product was eluted at 4-5% of methanol in dichloromethane to obtain the title compound (14.0 g, 40.4% yield).

LCMS (ESI): m/z 487.36 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 8.22 (d, J=2.0 Hz, 1H), 8.02 (s, 1H), 7.82-7.67 (m, 2H), 7.48-7.40 (m, 2H), 7.34 (s, 1H), 7.03-6.89 (m, 3H), 5.55-5.37 (m, 2H), 5.12 (s, 2H), 4.06 (t, J=5.6 Hz, 2H), 3.77 (s, 3H), 3.45 (t, J=5.3 Hz, 2H), 1.36 (s, 9H).

Step-2: Tert-butyl (E)-(4-((2-amino-4-carbamoyl-6-((4-methoxybenzyl)oxy)phenyl)amino) but-2-en-1-yl)carbamate

[Image Omitted]

In a round bottom flask, to a stirred solution of tert-butyl (E)-(4-((4-carbamoyl-2-((4-methoxybenzyl)oxy)-6-nitrophenyl)amino)but-2-en-1-yl)carbamate (Step 1, 14.00 g, 28.8 mmol) in methanol (500 ml) at 0° C., was added sodium dithionite (29.7 g, 144 mmol) in methanol:THF (50 ml, 1:1) followed by addition of ammonium hydroxide (53.8 ml, 345 mmol). The reaction mixture was allowed to warm at room temperature for 30 min. Reaction was monitored by TLC. The mixture was diluted with water (100 ml) and ethyl acetate (300 ml). Organic layer was separated and aqueous layer was extracted with ethyl acetate (300×3). The combined organic layer was dried over sodium sulphate and concentrated under vacuum to give the title compound as brown solid (11.5 g, 88% yield).

LCMS (ESI): m/z 457.50 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 7.62 (s, 1H), 7.40 (m, 2H), 7.06-6.80 (m, 5H), 5.67-5.41 (m, 2H), 5.01 (d, J=8.2 Hz, 2H), 4.70 (s, 1H), 3.77 (s, 3H), 3.53 (s, 3H), 3.34 (s, 2H), 1.37 (s, 9H).

Step-3: Tert-butyl (E)-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate

[Image Omitted]

In a round bottom flask, to a stirred solution of tert-butyl (E)-(4-((2-amino-4-carbamoyl-6-((4-methoxybenzyl)oxy)phenyl)amino)but-2-en-1-yl)carbamate (step 2, 11 g, 24.09 mmol) in dimethylformamide (100 ml), 1-ethyl-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (4.70 g, 24.09 mmol) in 1 ml of dimethylformamide was added at 0° C. and stirred for 30 min. To this reaction mixture, EDC (6.93 g, 36.1 mmol) and triethylamine (10.08 ml, 72.3 mmol) were added at 0° C. and stirred for 18 hr. After completion the mixture was concentrated under reduced pressure. Water was added, a resulting solid was filtered and dried to get the title compound (11.5 g, 18.62 mmol, 77% yield).

LCMS (ESI): m/z 618.46 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.86 (s, 1H), 8.00 (s, 1H), 7.67 (d, J=1.2 Hz, 1H), 7.53 (d, J=1.4 Hz, 1H), 7.50-7.46 (m, 2H), 7.38 (d, J=7.5 Hz, 1H), 7.08-6.87 (m, 5H), 6.61 (s, 1H), 5.67 (d, J=15.3 Hz, 1H), 5.38 (m, 1H), 5.22 (s, 2H), 4.85 (d, J=6.8 Hz, 2H), 4.60 (q, J=7.0 Hz, 2H), 3.78 (d, J=8.1 Hz, 5H), 3.44 (m, 2H), 2.17 (s, 4H), 1.32 (s, 9H).

Step 4: (E)-1-(4-aminobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide hydrochloride

[Image Omitted]

In a round bottom flask, tert-butyl (E)-(4-(5-carbamoyl-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)carbamate (step 3, 11.45 g, 18.54 mmol) was dissolved in dichloromethane (150.00 ml) and a methanol (~15 ml) was added to it. To the solution, was added hydrochloric acid (4 M in dioxane) (46.3 ml, 185 mmol) slowly. The reaction mixture was stirred at room temperature for 2 hr. Upon completion, the reaction mixture was concentrated and the solid obtained was triturated with diethyl ether to get the title compound (10.0 g, 97% yield).

LCMS (ESI): m/z 518.36 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 7.69 (d, J=1.2 Hz, 2H), 7.59 (d, J=1.4 Hz, 2H), 7.56-7.45 (m, 3H), 7.43-7.33 (m, 2H), 7.09-6.99 (m, 3H), 6.71 (m, 1H), 6.02-5.92 (m, 1H), 5.56-5.43 (m, 1H), 5.27 (s, 2H), 4.94 (d, J=5.6 Hz, 2H), 4.60 (q, J=6.8 Hz, 2H), 3.77 (m, 4H), 3.43-3.31 (m, 2H), 2.19 (d, J=4.4 Hz, 2H), 1.36 (m, 3H).

Step 5: (E)-1-(4-((4-carbamoyl-2-((4-methoxybenzyl)oxy)-6-nitrophenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide

[Image Omitted]

In a round bottom flask, (E)-1-(4-aminobut-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide hydrochloride (9.95 g, 17.96 mmol) and 4-chloro-3-((4-methoxybenzyl)oxy)-5-nitrobenzamide (step 4, 6.05 g, 17.96 mmol) was suspended in isopropyl alcohol (100 ml) and diisopropylethylamine (25.09 ml, 144 mmol) was added and the mixture heated at 115° C. in a sealed tube for 36 hrs. The progress of the reaction was monitored by LCMS. Upon completion (~10% of starting observed) the reaction mixture was concentrated under reduced pressure and was purified by column chromatography using ethyl acetate in hexane, to remove the starting compound. Then the desired compound was eluted at ~10-15% of methanol in dichloromethane to obtained the title compound (10.5 g, 71.5% yield).

LCMS (ESI): m/z 818.61 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 9.34 (br s, 2H), 8.05 (s, 1H), 7.34 (m, 3H), 7.17 (d, J=8.6 Hz, 1H), 6.89-6.81 (m, 1H), 6.76 (d, J=8.7 Hz, 1H), 4.92-4.72 (m, 2H), 4.55 (q, J=7.2 Hz, 2H), 4.00 (s, 2H), 3.77-3.68 (m, 3H), 3.59 (m, 8H), 3.35 (m, 8H), 3.12 (m, 6H), 2.16 (m, 2H).

Step 6: (E)-1-(4-((2-amino-4-carbamoyl-6-((4-methoxybenzyl)oxy)phenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide

[Image Omitted]

(E)-1-(4-((4-carbamoyl-2-((4-methoxybenzyl)oxy)-6-nitrophenyl)amino)but-2-en-1-yl)-2-(l-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide (step 7, 10 g, 12.23 mmol) was dissolved in 50 mL methanol. The solution was cooled to 0° C. and a solution of sodium dithionite (12.60 g, 61.1 mmol) in 25 ml water was added to it followed by 28% aqueous solution of ammonium hydroxide (22.85 ml, 147 mmol). The reaction mixture was allowed to warm to room temperature and stirred for 2 h after which TLC indicated completion of reaction. The volatiles were then concentrated on rotavapour and the crude product precipitated out. The solid obtained was stirred with 20% MeOH-DCM and then filtered to obtain the product (7.5 g, 78% yield).

LCMS (ESI): m/z 788.73 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 8.05 (s, 2H), 7.65 (m, 3H), 7.51 (t, J=1.6 Hz, 2H), 7.43-7.35 (m, 4H), 7.21 (m, 2H), 6.99 (m, 2H), 6.94-6.86 (m, 4H), 6.85-6.76 (m, 2H), 6.61 (dq, J=8.4, 4.1 Hz, 1H), 5.71 (m, 1H), 5.19-5.04 (m, 3H), 4.87-4.76 (m, 4H), 4.70-4.47 (m, 4H), 3.53 (m, 5H), 2.23-2.10 (m, 4H), 1.32-1.27 (m, 4H).

Step 7: (E)-1-(4-(5-carbamoyl-2-(1-(2-fluoroethyl)-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide

[Image Omitted]

(E)-1-(4-((2-amino-4-carbamoyl-6-((4-methoxybenzyl)oxy)phenyl)amino)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide (4.5 g, 5.71 mmol) was dissolved in 50 mL DMF and the solution cooled to 0° C. and a solution of 1-(2-fluoroethyl)-3-methyl-1H-pyrazole-5-carbonyl isothiocyanate (Intermediate 3, 1.22 g, 5.71 mmol) in 5 mL DMF was added to it. The reaction mixture was stirred at 0° C. for 30 minutes after which 3-(ethyliminomethylideneamino)-N,N-dimethylpropan-1-amine; hydrochloride (1.64 g, 8.57 mmol) was added to the reaction mixture followed by the addition of triethylamine (2.39 mL, 17.13 mmol). The reaction mixture was allowed to warm to RT and stirred overnight. The progress of the reaction was monitored by LCMS and upon completion, the reaction mixture was concentrated under reduced pressure and water was added. The solid precipitate obtained was filtered to get the crude product (2.3 g, 41.6% yield).

LCMS (ESI): m/z 967.70 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.91 (s, 2H), 7.98 (s, 2H), 7.74 (s, 2H), 7.40 (m, 4H), 7.14-6.97 (m, 3H), 6.82-6.64 (m, 4H), 6.55 (m, 2H), 5.48 (s, 2H), 4.99-4.73 (m, 10H), 4.70-4.45 (m, 3H), 3.61 (s, 6H), 3.33 (s, 4H), 2.12 (d, J=5.6 Hz, 6H), 1.35-1.23 (m, 4H)

Step 8: (E)-1-(4-(5-carbamoyl-2-(1-(2-fluoroethyl)-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazole-5-carboxamide

[Image Omitted]

(E)-1-(4-(5-carbamoyl-2-(1-(2-fluoroethyl)-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-((4-methoxybenzyl)oxy)-1H-benzo[d]imidazole-5-carboxamide (Product from Step 9, 2.2 g, 2.28 mmol) was dissolved in 50 mL DCM and trifluoroacetic acid (5.19 g, 45.5 mmol) was added and the reaction mixture stirred at room temperature for 16 h. The progress of the reaction was monitored by LCMS and upon completion, the reaction mixture was concentrated under reduced pressure and diethyl ether was added resulting in the precipitation of the product as a solid which was filtered and washed with ether (1.6 g, 97% yield).

LCMS (ESI): m/z 727.53 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 10.47 (s, 2H), 7.91-7.74 (m, 2H), 7.50-7.41 (m, 3H), 7.23 (s, 2H), 7.17-7.08 (m, 2H), 6.53 (m, 2H), 5.95 (m, 2H), 4.96 (s, 4H), 4.92-4.72 (m, 4H), 4.66 (t, J=5.1 Hz, 1H), 4.51 (q, J=7.1 Hz, 2H), 2.11 (m, 6H), 1.25 (t, J=7.1 Hz, 4H).

Step 9: (E)-15-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-1-(1-(2-fluoroethyl)-3-methyl-1H-pyrazole-5-carboxamido)-8-morpholino-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide formic acid salt (Compound 17)

[Image Omitted]

The title compound was prepared by following the procedure outlined for step 6 of Example 1 using (E)-1-(4-(5-carbamoyl-2-(1-(2-fluoroethyl)-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazol-1-yl)but-2-en-1-yl)-2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazole-5-carboxamide (step 8 product) and 4-(1,3-dichloropropan-2-yl)morpholine.

LCMS (ESI): m/z 852.5 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 8.01 (s, 2H), 7.77-7.71 (m, 2H), 7.66 (s, 2H), 7.41 (s, 2H), 6.57 (d, J=21.9 Hz, 2H), 5.53 (s, 2H), 4.99-4.86 (m, 4H), 4.86-4.74 (m, 2H), 4.72-4.59 (m, 3H), 4.53 (q, J=7.1 Hz, 2H), 4.21 (dd, J=10.4, 6.2 Hz, 2H), 3.76-3.62 (m, 4H), 3.15 (q, J=5.4 Hz, 1H), 2.83 (q, J=6.1, 4.7 Hz, 4H), 2.09-2.04 (m, 6H), 1.28-1.22 (m, 3H).

Example 18: (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-((4-methylpiperazin-1-yl)methyl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 18)

[Image Omitted]

The title compound as its dihydrochloride salt was prepared by following the procedure outlined for Example 1 using (E)-1,1′-(but-2-ene-1,4-diyl)bis(2-(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-7-hydroxy-1H-benzo[d]imidazole-5-carboxamide) and 1-(3-chloro-2-(chloromethyl)propyl)-4-methylpiperazine dihydrochloride.

LCMS (ESI): m/z 861.68 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 8.04 (brs, 2H), 7.95 (brs, 2H), 7.77 (s, 2H), 7.46 (brs, 2H), 6.57-6.55 (s, 2H), 5.42 (s, 2H), 4.92 (s, 6H), 4.51 (q, J=7.1 Hz, 6H), 4.38-4.36 (m, 2H), 2.90-2.88 (s, 3H), 2.60-2.53 (m, 2H), 2.50 (s, 3H), 2.06 (s, 6H), 1.55-1.53 (m, 1H), 1.26-1.23 (m, 6H).

Example 19: (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(1H-imidazol-1-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 19)

[Image Omitted]

LCMS (ESI): m/z 815.67 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 7.96 (s, 2H), 7.81 (s, 1H), 7.68 (s, 1H), 7.50-7.39 (m, 4H), 7.22 (s, 1H), 7.14 (s, 1H), 6.99 (s, 1H), 6.47 (s, 2H), 5.75-5.58 (m, 2H), 5.35 (s, 1H), 5.12 (d, J=9.8 Hz, 3H), 4.94-4.82 (m, 2H), 4.69-4.61 (m, 2H), 4.56-4.40 (m, 5H), 2.09 (s, 6H), 1.25-1.17 (m, 6H).

Example 20: (33R,35R,E)-12,62-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-11H,61H-2,5-dioxa-1,6(7,1)-dibenzo[d]imidazola-3(3,5)-pyrrolidinacyclodecaphan-8-ene-15,65-dicarboxamide (Compound 20)

[Image Omitted]

LCMS (ESI): m/z 789.91 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 10.44 (s, 1H), 9.14 (s, 1H), 8.13 (brs, 2H), 7.76 (s, 1H), 7.74 (s, 1H), 7.62 (brs, 1H), 7.47-7.40 (m, 3H), 6.59-6.57 (m, 2H), 6.00 (s, 1H), 5.81 (s, 1H), 5.58 (s, 1H), 5.26-5.07 (m, 2H), 4.90 (s, 2H), 4.61-4.51 (m, 6H), 4.32-4.22 (m, 1H), 3.94-3.84 (m, 1H), 2.98-2.87 (m, 1H), 2.06 (s, 3H), 2.01 (s, 3H), 1.29 (t, J=7.1 Hz, 6H).

Example 21: (33R,35R,E)-12,62-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-31-methyl-11H,61H-2,5-dioxa-1,6(7,1)-dibenzo[d]imidazola-3(3,5)-pyrrolidinacyclodecaphan-8-ene-15,65-dicarboxamide (Compound 21)

[Image Omitted]

To a stirred solution of (33S,35R,E)-12,62-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-11H,61H-2,5-dioxa-1,6(7,1)-dibenzo[d]imidazola-3(3,5)-pyrrolidinacyclodecaphan-8-ene-15,65-dicarboxamide (0.015 g, 0.019 mmol) [Example 20] in 3 mL of MeOH-DCM (1:2) was added formaldehyde (0.035 g, 0.38 mmol) and stirred for 2 h at 25° C. after which sodium triacetoxyborohydride (8.05 mg, 0.038 mmol) was added and stirred for 1 h at 25° C. The reaction was monitored by LCMS and after completion the volatiles were conc. under reduced pressure and water was added. The solid precipitated was filtered and dried to get the crude product which was purified by prep HPLC and converted to the hydrochloride salt.

LCMS (ESI): m/z 804.6 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 8.13 (s, 2H), 7.76 (m, 3H), 7.45 (s, 3H), 6.59 (s, 2H), 6.16-5.99 (m, 1H), 5.99-5.81 (m, 1H), 5.78-5.56 (m, 1H), 5.17-4.79 (m, 4H), 4.66-4.48 (m, 6H), 3.10 (s, 3H), 2.07 (s, 3H), 2.03 (s, 3H), 1.32-1.27 (m, 6H).

Example 22: (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(4-hydroxypiperidin-1-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 22)

[Image Omitted]

The title compound as its hydrochloride salt was prepared by following the procedure outlined for Example 11 using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and 2-(4-((tert-butyldimethylsilyl)oxy)piperidin-1-yl)propane-1,3-diol followed by deprotection of the TBS group and conversion to its salt.

LCMS (ESI): m/z 848.18 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 11.25 (s, 1H), 8.14 (s, 2H), 7.95 (d, J=6.8 Hz, 2H), 7.82-7.76 (m, 2H), 7.49 (s, 2H), 6.52 (s, 2H), 5.43-5.23 (m, 2H), 5.13 (d, J=11.2 Hz, 2H), 4.92-4.87 (m, 4H), 4.50-4.43 (m, 4H), 4.45-4.31 (m, 2H), 3.68-3.63 (m, 2H), 3.57-3.43 (m, 2H), 2.40-2.48 (m, 4H), 2.03 (s, 6H), 1.25 (t, J=6.9 Hz, 6H).

Example 23: (E)-8-(4-aminopiperidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 23)

[Image Omitted]

The title compound as its dihydrochloride salt was prepared by following the procedure outlined for Example 11 using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and tert-butyl (1-(1,3-dihydroxypropan-2-yl)piperidin-4-yl)carbamate followed by deprotection of the Boc group and conversion to its salt.

LCMS (ESI): m/z 847.30 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 11.85 (s, 2H), 8.41-8.11 (m, 6H), 7.82 (d, J=9.4 Hz, 2H), 7.47 (s, 2H), 6.52 (s, 2H), 5.26 (s, 2H), 4.91 (s, 4H), 4.49 (q, J=7.1 Hz, 4H), 4.38 (d, J=7.7 Hz, 2H), 4.08 (m, 2H), 3.91-3.52 (m, 6H), 2.19 (m, 4H), 2.03 (s, 6H), 1.23 (m, 6H).

Example 24: (S,E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diinden-8-yl)pyrrolidin-3-yl dihydrogen phosphate (Compound 24)

[Image Omitted]

Step 1: Synthesis of (S,E)-di-tert-butyl (1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diinden-8-yl)pyrrolidin-3-yl) phosphate

[Image Omitted]

To (S,E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(3-hydroxypyrrolidin-1-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 11, 150 mg, 0.180 mmol) in 2 mL DMF were added 1H-tetrazole (315 mg, 4.50 mmol) and di-tert-butyl diisopropylphosphoramidite (998 mg, 3.60 mmol) at 25° C. The mixture was allowed to stir at 25° C. for 2 hr. Progress of the reaction was monitored by LCMS. Upon complete consumption of the starting material, H₂O₂(30% aqueous solution 1.5 ml, 4.50 mmol) was added at room temperature and stirred for 30 min after which the reaction mixture was poured into 10 ml solution of water containing sodium bicarbonate and sodium thiosulphate. The product precipitated out which was then filtered and purified by preparative HPLC to afford the title compound (20 mg, 11% yield).

LCMS (ESI): m/z 1026.84 (M+H)⁺

Step 2: Synthesis of (S,E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diinden-8-yl)pyrrolidin-3-yl dihydrogen phosphate

[Image Omitted]

(S,E)-di-tert-butyl (1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diinden-8-yl)pyrrolidin-3-yl) phosphate (20 mg, 0.019 mmol) obtained from the previous step, was dissolved in 5 ml dichloromethane and 4 N HCl in dioxane (0.236 ml, 0.945 mmol) at room temperature and stirred for 2 h. The progress of the reaction was monitored by LCMS. After completion of the deprotection, the volatiles were concentrated under reduced pressure to get a solid which was triturated with diethyl ether to the hydrochloride salt of the title compound (15 mg, 80% yield).

LCMS (ESI): m/z 913.32 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) d 8.16 (s, 2H), 7.81 (d, J=5.8 Hz, 4H), 7.46 (s, 2H), 6.53 (s, 2H), 5.33 (d, J=10.2 Hz, 3H), 5.04-4.84 (m, 6H), 4.57-4.33 (m, 8H), 3.41-3.31 (m, 3H), 2.46-2.40 (m, 2H), 2.04 (s, 6H), 1.27-1.23 (m, 6H).

Example 25: (E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diinden-8-yl)piperidin-4-yl dihydrogen phosphate (Compound 25)

[Image Omitted]

The title compound as its hydrochloride salt was prepared by following the procedure outlined for Example 24 using (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(4-hydroxypiperidin-1-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 22)

LCMS (ESI): m/z 928.32 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.98 (s, 2H), 11.50 (s, 1H), 8.11 (s, 2H), 7.90 (d, J=26.8 Hz, 2H), 7.79 (d, J=4.3 Hz, 2H), 7.49 (s, 2H), 6.52 (s, 2H), 5.36-5.25 (m, 2H), 5.19-5.06 (m, 2H), 4.97-4.87 (m, 3H), 4.63-4.23 (m, 11H), 3.60 (s, 2H), 2.28-2.18 (m, 2H), 2.03 (s, 6H), 1.27-1.22 (m, 6H).

Example 26: (E)-8-(3-cyanopyrrolidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 26)

[Image Omitted]

Step 1: Synthesis of 1-(1,3-dichloropropan-2-yl)pyrrolidine-3-carbonitrile

[Image Omitted]

A mixture of 1,3-dichloropropan-2-one (2.62 g, 20.60 mmol) and pyrrolidine-3-carbonitrile (1.8 g, 18.72 mmol) in 30 ml THF was stirred for 2 hr at room temp. after which sodium triacetoxyborohydride (5.16 g, 24.34 mmol) was added and reaction mixture was stirred at room temperature for 12 h. After the completion of the reaction, it was quenched by the addition of water and extracted with DCM. The organic layer was washed with brine, dried over sodium sulphate and concentrated under reduced pressure to get crude product which was purified by flash column chromatography to get the product.

LCMS (ESI): m/z 207.01 (M+H)⁺

¹H NMR (400 MHz, Chloroform-d) δ 3.88-3.78 (m, 2H), 3.78-3.69 (m, 2H), 3.32-3.19 (m, 1H), 3.13-3.02 (m, 1H), 3.02-2.91 (m, 3H), 2.90-2.81 (m, 1H), 2.36-2.23 (m, 1H), 2.22-2.13 (m, 1H).

Step 2: Synthesis of dimethyl 5,5′-((2-(3-cyanopyrrolidin-1-yl)propane-1,3-diyl)bis(oxy))bis(4-chloro-3-nitrobenzoate)

[Image Omitted]

Methyl 4-chloro-3-hydroxy-5-nitrobenzoate (2.24 g, 9.66 mmol) and 1-(1,3-dichloropropan-2-yl)pyrrolidine-3-carbonitrile (1.00 g, 4.83 mmol) was dissolved in 50 ml of acetonitrile. DIPEA (5.06 ml, 29.0 mmol) and KI (0.802 g, 4.83 mmol) were added and the reaction mixture stirred at 80° C. for 12 h. The reaction mass was then concentrated under reduced pressure and purified by column chromatography to get dimethyl 5,5′-((2-(3-cyanopyrrolidin-1-yl)propane-1,3-diyl)bis(oxy))bis(4-chloro-3-nitrobenzoate) (1.3 g, 45.1% yield).

LCMS (ESI): m/z 596.83 (M+H)⁺

¹H NMR (400 MHz, Chloroform-_d) δ 8.10-8.06 (m, 2H), 7.85-7.80 (m, 2H), 4.55-4.44 (m, 4H), 4.00 (s, 6H), 3.70-3.65 (m, 1H), 3.37-3.28 (m, 1H), 3.25-3.19 (m, 1H), 3.14-3.03 (m, 3H), 2.40-2.19 (m, 2H).

Step 3: dimethyl 5,5′-((2-(3-cyanopyrrolidin-1-yl)propane-1,3-diyl)bis(oxy))bis(4-chloro-3-nitrobenzoate) obtained above was converted to (E)-8-(3-cyanopyrrolidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide hydrochloride salt (Compound 26) by following the procedure outlined in Step 4 to Step 8 of Example 11

[Image Omitted]

LCMS (ESI): m/z 843.18 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 8.10 (s, 2H), 7.85 (s, 2H), 7.80 (d, J=1.4 Hz, 2H), 7.49 (s, 2H), 6.54 (s, 2H), 5.11-4.73 (m, 6H), 4.56-4.42 (m, 6H), 4.35-4.21 (m, 8H), 2.48-2.44 (m, 2H), 2.04 (s, 6H), 1.26-1.23 (m, 6H).

Example 27: (E)-8-(3-aminopyrrolidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 27)

[Image Omitted]

The title compound as its dihydrochloride salt was prepared by following the procedure outlined for Example 26 using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and tert-butyl (1-(1,3-dichloropropan-2-yl)pyrrolidin-3-yl)carbamate followed by deprotection of the Boc group and conversion to its salt.

LCMS (ESI): m/z 833.30 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 11.85 (s, 2H), 8.41-8.11 (m, 6H), 7.82 (m, 2H), 7.47 (s, 2H), 6.52 (s, 2H), 5.26 (s, 2H), 4.91 (s, 4H), 4.49 (q, J=7.1 Hz, 4H), 4.38 (d, J=7.7 Hz, 2H), 4.08 (m, 2H), 3.91-3.52 (m, 6H), 2.19 (m, 4H), 2.03 (s, 6H), 1.23 (q, J=8.8, 7.8 Hz, 6H).

Example 28: (R,E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diinden-8-yl)pyrrolidin-3-yl dihydrogen phosphate (Compound 28)

[Image Omitted]

The title compound as its hydrochloride salt was prepared by following the procedure outlined for Example 24 using (R,E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(3-hydroxypyrrolidin-1-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 12)

LCMS (ESI): m/z 913.19 (M+H)⁺

1H NMR (400 MHz, DMSO-d6) δ 12.99 (s, 2H), 8.14 (m, 2H), 7.81 (d, J=6.7 Hz, 4H), 7.47 (s, 2H), 6.53 (s, 2H), 5.33 (d, J=10.8 Hz, 2H), 4.96 (m, 6H), 4.50 (q, J=7.0 Hz, 5H), 4.28-4.20 (m, 5H), 2.59-2.54 (m, 4H), 2.04 (s, 6H), 1.24 (d, J=1.6 Hz, 6H).

Example 29: (E)-8-(4-cyanopiperidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 29)

[Image Omitted]

The title compound as its hydrochloride salt was prepared by following the procedure outlined for Example 26 using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and 1-(1,3-dichloropropan-2-yl)piperidine-4-carbonitrile.

LCMS (ESI): m/z 857.05 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 8.00 (s, 2H), 7.63 (d, J=28.0 Hz, 4H), 7.37 (s, 2H), 6.52 (s, 2H), 5.51 (s, 2H), 4.90 (m, 4H), 4.54 (m, 6H), 4.20 (s, 2H), 3.19 (s, 1H), 2.92 (m, 4H), 2.76 (m, 2H), 2.07 (s, 6H), 1.97 (s, 2H), 1.88-1.74 (m, 2H), 1.25 (d, J=8.3 Hz, 6H).

Example 30: (E)-8-(azetidin-3-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 30)

[Image Omitted]

The title compound as its hydrochloride salt was prepared by following the procedure outlined for Example 11 using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and tert-butyl 3-(1,3-dihydroxypropan-2-yl)azetidine-1-carboxylate followed by deprotection of the Boc group and conversion to its salt.

LC-MS (ESI): m/z 804.29 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 9.22 (s, 1H), 9.07 (s, 1H), 8.09 (s, 2H), 7.75 (d, J=1.2 Hz, 2H), 7.68 (s, 2H), 7.43 (s, 2H), 6.54 (s, 2H), 5.42 (s, 2H), 4.91 (s, 4H), 4.51 (q, J=7.1 Hz, 4H), 4.43 (m, 2H), 4.23-4.05 (m, 6H), 3.18-3.06 (m, 1H), 2.18 (d, J=7.1 Hz, 1H), 2.05 (s, 6H), 1.25 (t, J=6.9 Hz, 6H).

Example 31: (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(piperidin-4-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 31)

[Image Omitted]

The title compound as its hydrochloride salt was prepared by following the procedure outlined for Example 11 using methyl 4-chloro-3-hydroxy-5-nitrobenzoate and tert-butyl 4-(1,3-dihydroxypropan-2-yl)piperidine-1-carboxylate followed by deprotection of the Boc group and conversion to its salt.

LCMS (ESI): m/z 832.17 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 8.84 (s, 1H), 8.71 (m, 1H), 8.07 (s, 2H), 7.73 (d, J=1.2 Hz, 2H), 7.65 (d, J=1.4 Hz, 3H), 7.42 (s, 2H), 6.54 (s, 2H), 5.44 (s, 2H), 4.92 (m, 4H), 4.51 (q, J=7.3 Hz, 6H), 4.23-4.12 (m, 2H), 3.37 (d, J=7.0 Hz, 2H), 2.96-2.86 (m, 2H), 2.25-2.12 (m, 2H), 2.06 (s, 8H), 1.81 (m, 2H), 1.25 (t, J=7.0 Hz, 6H).

Example 32: (E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diinden-8-yl)pyrrolidine-3-carboxylic acid (Compound 32)

[Image Omitted]

Step 1: Synthesis of methyl (E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diinden-8-yl)pyrrolidine-3-carboxylate

[Image Omitted]

(E)-8-(3-cyanopyrrolidin-1-yl)-1,15-bis(I-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (50 mg, 0.06 mmol) [Compound 26] was dissolved in 5 mL MeOH and 4 N HCl in Dioxane (1.48 ml, 5.93 mmol) was added to it and stirred for 4 h at room temperature. The reaction was monitored by LCMS, and after conversion to the ester, the reaction mixture was concentrated under reduced pressure and washed with diethyl ether to obtain the title compound. (40 mg, 77% yield)

LCMS (ESI): m/z 876.18 (M+H)⁺

Step 2: (E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diinden-8-yl)pyrrolidine-3-carboxylic acid (Compound 32)

[Image Omitted]

methyl (E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diinden-8-yl)pyrrolidine-3-carboxylate hydrochloride (Step 1, 30 mg, 0.033 mmol) was dissolved in a 6 mL mixture of MeOH, THF and Water (1:1:1). Lithium hydroxide hydrate (6.90 mg, 0.164 mmol) was added and the reaction mixture stirred at RT for 15 h. The reaction was monitored by LCMS, and after complete hydrolysis the reaction mixture was conc. under reduced pressure and water was added. The pH of the mixture was adjusted to acidic by the addition of 2 N HCl, and the solid precipitated out was filtered and dried to get the crude product which was purified by prep HPLC to obtain the title product. (10 mg, 34% yield)

LCMS (ESI): m/z 862.18 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.99 (brs, 1H), 11.55 (brs, 1H), 11.24 (brs, 1H), 8.06 (brs, 2H), 7.80 (d, J=2.8 Hz, 4H), 7.49 (brs, 2H), 6.54 (s, 2H), 5.33 (s, 2H), 5.07-4.80 (m, 6H), 4.61-4.35 (m, 6H), 4.27-4.13 (m, 1H), 4.13-3.99 (m, 1H), 3.95-3.82 (m, 2H), 3.45-3.28 (m, 2H), 2.04 (s, 6H), 1.25 (t, J=7.6 Hz, 6H).

Example 33: (E)-1-(4,12-dicarbamoyl-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diinden-8-yl)piperidine-4-carboxylic acid (Compound 33)

[Image Omitted]

The title compound was prepared from (E)-8-(4-cyanopiperidin-1-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 29) by following the procedure outlined in Example 32.

LCMS (ESI): m/z 876.31 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 7.90 (brs, 2H), 7.49 (d, J=1.2 Hz, 2H), 7.35 (s, 2H), 7.11 (brs, 2H), 6.38 (s, 2H), 5.33 (s, 2H), 5.00 (m, 2H), 4.83 (d, J=15.7 Hz, 2H), 4.53 (d, J=7.2 Hz, 6H), 4.05-3.99 (m, 4H), 3.00 (d, J=1.9 Hz, 2H), 2.06 (s, 6H), 1.85-1.74 (m, 2H), 1.64-1.56 (m, 2H), 1.22-1.15 (m, 6H).

Example 34: (E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(1-methylazetidin-3-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 34)

[Image Omitted]

The title compound as its formate salt was prepared by following the reductive amination procedure outlined in Example 21 using (E)-8-(azetidin-3-yl)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 30) and purification of the crude product by prep HPLC.

LCMS (ESI): m/z 818.17 (M+H)⁺

¹H NMR (400 MHz, DMSO-d₆) δ 12.90 (brs, 2H), 8.02 (brs, 2H), 7.73 (d, J=1.3 Hz, 2H), 7.56 (d, J=1.4 Hz, 2H), 7.40 (s, 2H), 6.53 (s, 2H), 5.41 (s, 2H), 4.89 (s, 4H), 4.51 (q, J=7.1 Hz, 4H), 4.32 (m, 2H), 4.14 (m, 2H), 3.60 (t, J=7.3 Hz, 2H), 3.21 (t, J=7.3 Hz, 2H), 2.33 (s, 3H), 2.06 (s, 6H), 1.27-1.23 (m, 6H).

Example 35: (R,E)-1,15-bis(1-ethyl-3-methyl-1H-pyrazole-5-carboxamido)-8-(3-methoxypyrrolidin-1-yl)-8,9,16,19-tetrahydro-7H-6,10-dioxa-2,14,15a,19a-tetraazacyclopentadeca[3,2,1-cd:8,9,10-c′d′]diindene-4,12-dicarboxamide (Compound 35)