The “administration” refers to providing the xanthone compound represented by Chemical Formula 1, the optical isomer thereof, or the pharmaceutically acceptable salt thereof to a subject by any appropriate method. At this time, the subject may specifically mean an animal, more specifically a mammal that may typically exhibit beneficial effects on prevention or treatment using the xanthone compound represented by Chemical Formula 1, the optical isomer thereof, or the pharmaceutically acceptable salt thereof, and much more specifically a primate such as a human, but is not limited thereto.

The pharmaceutical composition further includes one or more active ingredients (active agents) that exhibit the same as or similar to the xanthone compound represented by Chemical Formula 1, the optical isomer thereof, or the pharmaceutically acceptable salt thereof without departing from impairing the effect of the present invention, thereby expecting an effective synergistic effect of the active ingredients.

Another aspect of the present invention relates to a food composition for preventing or improving cancer including at least one selected from the group consisting of the xanthone compound represented by Chemical Formula 1 described above, an optical isomer thereof, and a pharmaceutically acceptable salt thereof. At this time, the aforementioned contents may be applied in the same manner to specific details about the type of cancer.

The food composition may be used as health functional food, and at this time, the “health functional food” refers to food produced and processed using raw materials or ingredients with functionality, which are useful for the human body according to the Art on Health Functional Foods No. 6727, and the “functionality” means intake for adjusting nutrients for the structures and functions of the human body or obtaining a useful effect on health applications such as physiological actions.

The food composition may further include common food additives disclosed in the art, and at this time, the suitability as the “food additive” is determined by the specifications and standards for the corresponding item in accordance with the general rules of the Food Additive Codex, general test methods, and the like approved by the Food and Drug Administration, unless otherwise specified.

The content of the xanthone compound represented by Chemical Formula 1, the optical isomer thereof, or the pharmaceutically acceptable salt thereof included in the food composition is not particularly limited, but may be included in an amount of 0.01 to 95 wt %, preferably 1 to 80 wt %, based on 100 wt % of the total composition, for the purpose of preventing or improving cancer. In addition, the food composition may be produced and processed in the form of tablets, capsules, powders, granules, liquids, pills, beverages, and the like, for the purpose of preventing or improving cancer-related diseases.

Another aspect of the present invention relates to a method for preventing or treating cancer-related diseases including administering at least one selected from the group consisting of the xanthone compound represented by Chemical Formula 1 described above, an optical isomer thereof, and a pharmaceutically acceptable salt thereof to a subject in need thereof. At this time, the cancer-related disease is not particularly limited thereto, but may be at least one selected from the group consisting of pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphocytic leukemia, basal cell cancer, ovarian cancer, ovarian epithelial cancer, ovarian germ cell cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colorectal cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, diffuse large B-cell lymphoma, ampullar of vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, sinonasal cancer, non-small cell lung cancer, non-Hodgkin lymphoma, tongue cancer, astrocytoma, small cell lung cancer, childhood brain cancer, childhood lymphoma, childhood leukemia, small intestine cancer, meningioma, esophageal cancer, glioma, neuroblastoma, renal pelvis cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, stomach cancer, gastric carcinoid, gastrointestinal cancer, Wilms cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational trophoblastic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, mesothelioma, rectal cancer, rectal carcinoid, vaginal cancer, spinal cord cancer, acoustic neuroma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsillar cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, and thymic cancer, and preferably at least one selected from the group consisting of colon cancer, stomach cancer, liver cancer, kidney cancer, prostate cancer, and breast cancer.

Advantageous Effects

According to the present invention, the present invention relates to a novel xanthone compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, and has the advantage of having an effect of preventing, treating or improving various cancer cells by exhibiting cytotoxicity against cancer cell lines to thereby inhibit cell growth of cancer cells.

Further, the method for preparing the novel xanthone compound, the optical isomer thereof, or the pharmaceutically acceptable salt thereof of the present invention has an advantage capable of preparing the xanthone compound represented by Chemical Formula 1, the optical isomer thereof, or the pharmaceutically acceptable salt thereof having an excellent effect of preventing, treating, or improving various cancer cells.

In addition, the pharmaceutical composition of the present invention includes at least one selected from the group consisting of the xanthone compound described above, the optical isomer thereof, or the pharmaceutically acceptable salt thereof, thereby having an advantage of having an excellent effect of preventing or treating cancer.

In addition, the food composition of the present invention includes at least one selected from the group consisting of the xanthone compound described above, the optical isomer thereof, or the pharmaceutically acceptable salt thereof, thereby having an advantage of having an excellent effect of preventing or improving cancer.

DESCRIPTION OF DRAWINGS

FIG. 1 is a diagram illustrating COSY and HMBC correlations of a compound represented by Chemical Formula 1-1 of the present invention.

FIG. 2 is a diagram illustrating COSY and HMBC correlations of a compound represented by Chemical Formula 1-2 of the present invention.

FIG. 3 is a diagram illustrating ¹H proton spectra of the compounds represented by Chemical Formulas 1-1 and 1-2 of the present invention.

FIG. 4 is a diagram illustrating a volume change of cancer cells in Experimental Example 3 of the present invention.

FIG. 5 is a diagram illustrating the weight of cancer cells in Experimental Example 3 of the present invention.

FIG. 6 is a photograph showing cancer cells in Experimental Example 3 of the present invention, in which (a) means a control group, (b) means Example 1, (c) means Example 2, and (d) means a comparison group.

MODES FOR THE INVENTION

Hereinafter, the present invention will be described in detail by Examples and Experimental Examples.

However, the following Examples are only illustrative of the present invention, and it will be apparent to those skilled in the art that various changes and modifications may be made within the scope and spirit of the present invention, and it is natural that these variations and modifications are within the scope of the appended claims. In the following Examples and Comparative Examples, “%” and “part” representing the content are based on a mass basis, unless otherwise specified.

Experimental Devices

- 1D (¹H and ¹³C), 2D (COSY, ROESY, HSQC, HMBC) NMR spectra: Bruker 600 MHz spectrometer
- UV spectrum: Shimadzu UV-1650PC spectrophotometer
- IR spectrum: JASCO FT/IR-4100 spectrophotometer
- Optical rotation: Rudolph Research Analytical (Autopol III) polarimeter
- High resolution (HR) ESIMS: Shimadzu LC/MS-IT-TOF mass spectrometer
- HPLC: RI-101 (Shodex) detector and PrimeLine Binary pump
- Analytical HPLC: ODS column (YMC-Pack-ODS-A, 250*4.6 mm, i.d, 5 μm)

Example: Production of Compounds

Isolation and Mass Culture of Actinomadura geliboluensis 188DD-028 Strain

A strain was isolated from a sponge sample collected from the coast of Dokdo, an island on the east coast of Korea. Based on the result of 16S rRNA gene sequence analysis (GenBank registration No. OK287077), the strain was identified as Actinomadura geliboluensis 188DD-028.

The strain Actinomadura geliboluensis 188DD-028 isolated in this way was cultured in a Bennett (BN) agar medium at 28° C. for 7 days, and thereafter, the strain was inoculated into a flask containing 50 mL of BN broth (containing 10 g glucose, 1 g yeast extract, 2 g tryptone, 1 g beef extract, 5 g glycerol, and 32 g NaCl in 1 L distilled water) and cultured at 28° C. for 7 days while stirring at 130 rpm. 50 mL of each culture medium was inoculated into a 2 L flask containing a 10% BN liquid culture medium (containing 1 g glucose, 0.1 g yeast extract, 0.2 g tryptone, 0.1 g beef extract, 0.5 g glycerol, and 32 g NaCl in 1 L distilled water) and a salt-free 100% BN liquid culture medium (containing 10 g glucose, 1 g yeast extract, 2 g tryptone, 1 g beef extract, 5 g glycerol, and 0 g NaCl in 1 L of distilled water) and cultured for 10 days under the same conditions.

The Actinomadura geliboluensis 188DD-028 was deposited on Nov. 10, 2021, and received deposit number KCTC 14764BP.

Isolation of Compounds

The 10% BN, and salt-free 100% BN culture media were extracted twice with the same amount of ethyl acetate, and concentrated under reduced pressure to obtain crude extracts. Thereafter, fractions soluble in a methylene chloride (MC) solvent were collected and subjected to chromatography, and among them, an acetone fraction was concentrated under reduced pressure. As a result, 2 mg of the compound represented by Chemical Formula 1-1 (Example 1) was obtained in the 10% BN culture medium, and 2 mg of the compound represented by Chemical Formula 1-2 (Example 2) was obtained in the salt-free 100% BN culture medium.

Compound represented by Chemical Formula 1-1 (Example 1): 8,15,16-trihydroxy-10,11-dimethoxy-3-methyl-5-((3,4,5-trimethoxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-3,4-dihydropyrano[4′,3′:6,7]naphtho[1,2-b]xanthene-1,14-dione]

Orange powder;

[
α
]

D
25

−60.0(c 0.1, CHCl₃); HRESIMS m/z 691.2027 [M+H]⁺(calcd for 691.2027, C₃₆H₃₅O₁₄).

Compound represented by Chemical Formula 1-2 (Example 2): 8,15,16-trihydroxy-10,11-dimethoxy-3-methyl-5-((3,4,5-trimethoxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-3,4,6,7-tetrahydropyrano[4′,3′:6,7]naphtho[1,2-b]xanthene-1,14-dione]

Brown powder;

[
α
]

D
25

−60.0(c 0.1, CHCl₃); HRESIMS m/z 695.2340 [M+H]⁺(calcd for 695.2340, C₃₆H₃₉O₁₄).

The ¹H NMR (CD₃OD, 600 MHz) and ¹³C NMR (CD₃OD, 125 MHz) results of each of the compounds represented by Chemical Formulas 1-1 (Example 1) and 1-2 (Example 2) were disclosed in Table 1 below.

TABLE 1

Example 1 (AG028-02)
Example 2 (AG028-03)

Position
δ_C
TYPE
δ_H(J, Hz)
δ_C
TYPE
δ_H(J, Hz)

1
170.1
C
—
170.5
C
—

3
75.9
CH
4.67, m
75.9
CH
4.55, br

4
30.3
CH₂
2.79, dd
39.5
CH₂
2.64, 3.44 br

(16.8, 5.4)

3.45, br

3.72, dd

(16.8, 3.0)

4a
131.5
C
—
131.2
C
—

5
139.7
C
—
142.4
C
—

5a
138.4
C
—
132.4
C
—

6
127.4
CH
8.55, d (9.0)
22.8
CH₂
2.15, 3.35, brs

7
124.6
CH
8.26, d (9.0)
22.8
CH₂
2.15, 3.35, brs

7a
129.6
C
—
104.6
C
—

8
178.5
C
—
160.4
C
—

8a
153.0
C
—
135.7
C
—

10
149.5
C
—
150.2
C
—

11
148.6
C
—
149.2
C
—

12
151.2
C
—
149.6
C
—

13
119.5
CH
7.37, d (9.6)
120.6
CH
7.29, d (9.6)

14
114.3
CH
7.44, d (9.6)
112.6
CH
7.16, d (9.6)

14a
120.9
C
—
115.8
C
—

15
173.0
C
—
182.1
C
—

15a
121.1
C
—
106.8
C
—

16
181.5
C
—
156.9
C
—

16a
136.6
C
—
120.1
C
—

16b
119.7
C
—
113.6
C
—

17
160.0
C
—
152.0
C
—

17a
104.5
C
—
107.3
C
—

3-CH3
20.9
CH₃
1.55, d (6.6)
20.9
CH₃
1.48, d (6.6)

11-OCH3
62.0
CH₃
4.01, s
61.8
CH₃
3.92, s

12-OCH3
56.8
CH₃
3.93, s
57.1
CH₃
3.86, s

17-OH
—
—
12.7, brs
—
—
13.1, brs

1′
105.0
CH
4.48, d (8.4)
106.9
CH
4.29, br

2′
84.4
CH
3.32, dd
84.4
CH
3.16, dd

(8.4, 9.0)

(7.8, 9.0)

3′
86.5
CH
3.16, dd
86.5
CH
3.07, dd

(8.4, 9.0)

(9.0, 9.0)

4′
85.1
CH
2.82, dd
85.2
CH
2.74, dd

(8.4, 9.0)

(9.0, 9.0)

5′
71.4
CH
3.01, m
71.1
CH
2.98, m

2′-OCH₃
61.1
CH₃
3.79, s
61.0
CH₃
3.66, s

3′-OCH₃
61.0
CH₃
3.67, s
61.0
CH₃
3.59, s

4′-OCH₃
60.7
CH₃
3.54, s
60.7
CH₃
3.48, s

5′-CH₃
17.5
CH₃
1.14, d (6.0)
17.6
CH₃
1.06, d (6.0)

8-OH
—
—
—
—
—
5.3, brs

16-OH
—
—
—
—
—
11.7, brs

Experiment Example 1: Determination of Planar Structures of Compounds

The planar structures of Examples 1 and 2 were determined by detailed analyses of 2D NMR data and ¹H-¹H COSY and HMBC spectra, respectively (see FIGS. 1 and 2).

In the ¹H NMR spectrum of Example 1, it could be seen that there were 4 aromatic protons (δ_H8.55, 8.26, 7.44, 7.37), 1 exchangeable proton (δ_H12.7), 6 oxygenated methines (δ_H4.67, 4.48, 3.32, 3.16, 3.01, 2.82), 5 methoxy groups (δ_H4.01, 3.93, 3.79, 3.67, 3.54), 1 methylene (δ_H2.79, 3.72), and 2 methyl groups (δ_H1.55, 1.14). As a result of analyzing the ¹³C NMR data and HSQC spectrum of Example 1, it was found that there were 4 carbonyl carbons, 18 aromatic carbons (4 protonated carbons), 6 methine carbons, 1 methylene carbon, and 7 methyl carbons. The planar structure of Example 1 was determined by 2D NMR (¹H-¹H COSY, HSQC, HMBC) data analysis (see FIG. 1). It was confirmed that there were 4 partial structures through COSY correlations of H-13/H-14, H-6/H-7, H-3-CH3/H-3/H-4 and H-1′/H-2′/H-3′/H-4′/H-5′/H-5′-CH₃. First, the structure of A part was determined by HMBC correlations between H-13 and C-11, C-12, H-14 and C-10, C-12, C-14a, C-15, H-11-O—CH₃and C-11, and H-12-OCH₃and C-12 (see FIG. 1). Through the H-1′/H-2′/H-3′/H-4′/H-5′/H-5′-CH₃spin system and the HMBC correlation between H-1′ and C-5′, a rhamnose structure was confirmed, and it was determined by the HMBC correlations that 2′, 3′, and 4′-OH were substituted with OCH₃. Next, the structure of B part was confirmed by using remaining two spin systems and HMBC correlation analysis of H-6 and C-5, C-7a, C-16b, H-7 and C-5a, C-8, C-16a, H-4 and C-5, C-17a, 17-OH and C-16b, C-17, C-17a, and by the HMBC correlation between H-1′ and C-5, it was confirmed that the rhamnose structure was linked to the B part structure through ester bonds. Because there were no HMBC signals related to non-protonated aromatic carbons, the linkage of the A and B part structures could not be determined clearly by HMBC correlations. In order to confirm strong signals of non-protonated aromatic carbons, glucose substituted with ¹³C was used in the cultivation of the producing strain to obtain ¹³C-(Example 1). By data analysis using ¹³C-(Example 1), C¹³-C¹³COSY correlations of C-1/C-17a, C-4/C-4a, C-8/C-8a, and C-15/C-15a/C-16/C-16a were confirmed (see FIG. 1). Based on the C¹³-C¹³COSY correlation analysis, the linkage of A and B part structures and the planar structure were confirmed, and as a result, the structure of Example 1 was determined as a novel xanthone-based compound.

The ¹H NMR spectrum of Example 2 showed two additional exchangeable protons (δ_H11.7, 5.3) and two methylenes (δ_H2.15, 3.35*2) compared to Example 1, and similar to the data of Example 1 except that two aromatic protons (δ_H8.55, 8.26) disappeared (see FIG. 3). By analyzing the ¹³C NMR data and HSQC spectrum of Example 2, it was confirmed that signals for C-6 and C-7 unsaturated carbons were replaced with two saturated carbons at δC 22.8, respectively. The disappearance of two carbonyl carbons and the appearance of two non-hydrogen aromatic carbons were also confirmed by the detailed analysis of NMR data. Accordingly, by considering chemical shift values and 2D NMR data of Example 2 that were almost the same as those of Example 1, it was revealed that the structure of Example 2 had a similar structure to Example 1. Example 2 had the same planar structure as Example 1 (see FIG. 2), except that C-6/C-7, C-8, and C-16 were reduced and substituted with two methylenes, C-8-OH, and C-16-OH, respectively.

Experiment Example 2: Cytotoxicity Test Against Cancer Cell Lines

Human cancer cell lines HCT-15 (colon cancer), NUGC-3 (stomach cancer), NCI-H23 (liver cancer), ACHN (kidney cancer), PC-3 (prostate cancer), and MDA-MB-231 (breast cancer) were purchased and used from American Type Culture Collection (Manassas, VA).

Each of the cancer cell lines was cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS) at 37° C. in a humidified atmosphere of 5% CO₂. Growth inhibition was evaluated for human cancer cell lines using a sulforhodamine B (SRB) assay. The cells were dispensed in a 96-well plate at a density of 8000 cells/well, and the next day, the cells were treated with each of Examples 1 and 2 (0.1% DMSO as vehicle control), and a positive control group was treated with Adriamycin. The cells were cultured for 48 hours, and then each culture medium was fixed with 50% trichloroacetic acid (50 μg/mL) and stained using 0.4% sulforhodamine B in 1% acetic acid. An unbound dye was washed with 1% acetic acid, and the dye bound to a protein was extracted with 10 mM Tris base (pH 10.5) to determine the optical density. Absorbance at 540 nm was determined using a VersaMax microplate reader (Molecular Devices, LLC, Sunnyvale, CA, USA). GI₅₀values were calculated using GraphPad Prism 4.0 software (GraphPad Software, Inc., San Diego, CA, USA), and the results are described in Table 2 below.

TABLE 2

GI₅₀(μM)^a

Example 1
Example 2

Cell line
(AG028-02)
(AG028-03)
Positive control^b

HCT-15
0.059
0.005
0.153

NUGC-3
0.017
0.003
0.145

NCI-H23
0.029
0.004
0.139

ACHN
0.032
0.005
0.151

PC-3
0.051
0.003
0.137

MDA-MB-231
0.036
0.004
0.146

^aGI₅₀value was a concentration value corresponding to 50% growth inhibition/

^bAdriamycin (ADR)

Referring to Table 2, it was confirmed that with respect to the two novel xanthone compounds of the present invention, the compound represented by Chemical Formula 1-1 showed GI₅₀values ranging from 0.017 to 0.059 μM, and the compound represented by Chemical Formula 1-2 showed GIso values ranging from of 0.003 to 0.005 μM. That is, it was confirmed that the novel two compounds of the present invention showed excellent activity against various types of cancer cell lines.