Novel compound or its salt and antitumor agent containing the same as an active ingredient

Abstract

Provided are: a novel compound having improved in-vivo efficacy and antitumor activity of a curcumin derivative, or a salt thereof; and an antitumor activator containing the same as an active ingredient. The present invention is a compound represented by the following formula or a salt thereof. R1-R4 are each a hydrogen atom or a substituent selected from among a C1-4 lower alkyl group, a hydroxy C1-4 lower alkyl group, a C1-4 lower alkoxy C1-4 lower alkyl group, and a C1-4 lower alkoxy C1-4 lower alkoxy C1-4 lower alkyl group, and R1-R4 may be the same as or different from each other. n is 1-6. Sugar is a monosaccharide or a disaccharide.

Description

Technical Field

[0001] The present application relates to a novel compound or a salt thereof and an antitumor agent containing the same as an active ingredient.

Background Art

[0002] Curcumin contained in spices and the like is known to have various pharmacological actions including antitumor activity. For example, the pharmacological actions of curcumin include antitumor activity, anti-inflammatory activity, anti-heart failure activity, antibacterial activity, radiation protection effect and the like. In addition, curcumin is an edible spice and is also known to have low toxicity.

[0003] The present inventors have previously disclosed in Patent Document 1 a curcumin derivative having enhanced antitumor activity while maintaining low toxicity. Specifically, Patent Document 1 discloses a novel compound, a predetermined bis(arylmethylene)acetone compound or a salt thereof, and an expression inhibitor of Ki-Ras, ErbB2, c-Myc or CyclinD1, a β-catenin degrading agent, an expression enhancer of p53, an anticancer agent or a carcinogenesis preventive agent containing these as active ingredients.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0005] As described in Patent Document 1, the present inventors have succeeded in developing a curcumin derivative having enhanced antitumor activity of curcumin. However, the curcumin derivative of Patent Document 1 has low water solubility and could not improve in vivo efficacy, so its pharmaceutical use has been limited.

[0006] Therefore, in view of the above circumstances, an object of the present disclosure is to provide a novel compound or a salt thereof having improved in vivo efficacy and antitumor activity of curcumin derivatives, and an antitumor active agent containing these as active ingredients.

Means for Solving the Problems

[0007] As one aspect for solving the above problems, the present disclosure provides a compound represented by the following general formula or a salt thereof.

[0008]

Chemical formula

[0009] Here, R 1 ~R 4 are substituents selected from a hydrogen atom, a C 1-4 lower alkyl group, a hydroxy C 1-4 lower alkyl group, a C 1-4 lower alkoxy C 1-4 lower alkyl group, and a C 1-4 lower alkoxy C 1-4 lower alkoxy C 1-4 lower alkyl group. R 1 ~R 4 may be the same or different. n is 1 to 6. Sugar is a monosaccharide or a disaccharide.

[0010] Further, the present disclosure provides an antitumor active agent containing the above compound or a salt thereof as an active ingredient. The antitumor active agent may be an antitumor active agent for gastric cancer, colorectal cancer, pancreatic cancer, malignant mesothelioma, cutaneous T-cell lymphoma, may be an antitumor active agent for pancreatic cancer, malignant mesothelioma, cutaneous T-cell lymphoma, or may be a pancreatic cancer antitumor active agent.

Effects of the Invention

[0011] Since the compound of the present disclosure or a salt thereof has improved water solubility compared to the curcumin derivative of Patent Document 1, its in vivo efficacy is enhanced. Therefore, it can be used for a wide range of pharmaceutical applications. Further, the compound of the present disclosure or a salt thereof also has improved antitumor activity compared to the curcumin derivative of Patent Document 1. Furthermore, the safety of the compound of the present disclosure or a salt thereof has also been confirmed in experiments using mice.

Brief Description of the Drawings

[0012]

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Mode for Carrying Out the Invention

[0013] The inventors of the present invention earnestly studied in order to improve the in vivo effectiveness (water solubility) of the curcumin derivative (diaryl pentanoid derivative: DPA) of Patent Document 1. As a result, SuDPA in which a saccharide (Sugar) is added to DPA via a linker was newly synthesized. Then, as a result of conducting various experiments on the synthesized SuDPA, it was found that SuDPA had significantly improved water solubility compared to DPA, and also had improved antitumor activity. Furthermore, the cancer molecular targeting property of DPA was also maintained in SuDPA. In addition, it was also found to be low in toxicity. Based on the above findings, the compound or its salt of the present disclosure was invented. Hereinafter, the compound or its salt of the present disclosure will be described.

[0014] The present disclosure provides a compound represented by the following general formula or a salt thereof.

[0015]

Chemical formula

[0016] Here, R 1 ~R 4 is a substituent selected from a hydrogen atom, a C 1-4 lower alkyl group, a hydroxy C 1-4 lower alkyl group, a C 1-4 lower alkoxy C 1-4 lower alkyl group, and a C 1-4 lower alkoxy C 1-4 lower alkoxy C 1-4 lower alkyl group. R 1 ~R4 may be the same or different. R 1 ~R 4 is C 1-4 lower alkoxy C 1-4 may be a lower alkyl group or a methoxymethyl group.

[0017] Here, the C in the above-described substituents 1-4 carbon chain may be linear or branched. Further, the carbon chain may be substituted with a halogen atom or the like.

[0018] n indicating the length of the linker moiety (vinyl alcohol moiety) connecting the DPA moiety and the saccharide moiety is 1 to 6. From the viewpoint of improving water solubility, n may be 2 to 5, may be 3 to 4, or may be 4.

[0019] Sugar is a monosaccharide or a disaccharide, and these deoxy sugars are also included. Examples of the monosaccharide include glucose, galactose, mannose, and deoxyglucose. Examples of the disaccharide include lactose and maltose. These saccharides are concepts including optical isomers. For example, Sugar may be D, L-glucose, D-galactose, D-mannose, D-lactose, D-maltose, and deoxyglucose. From the viewpoint of improving water solubility, Sugar may be D-galactose, L-glucose, D-mannose, D-lactose, D-maltose. From the viewpoint of improving antitumor activity, Sugar may be D-mannose, D-lactose, and D-maltose. From the viewpoint of synthesis yield, Sugar may be D-lactose. Here, the position of Sugar bonded to the adjacent triazole group is not particularly limited, but from the viewpoint of ease of synthesis, it may be the carbon at the 1-position.

[0020] The salts of the compounds of the present disclosure are not particularly limited, and examples thereof include sodium salts, potassium salts, calcium salts, or magnesium salts of the compounds of the present disclosure.

[0021] The compound of the present disclosure or a salt thereof has an improved water solubility and antitumor activity compared to the curcumin derivative of Patent Document 1 due to a structure in which a saccharide is added to the DPA site via a linker. The compound of the present disclosure or a salt thereof has an inhibitory effect on NF-kB, pSTAT3, β-catenin, and / or fatty acid synthase, thereby exhibiting antitumor activity. Further, the compound of the present disclosure or a salt thereof further has an angiogenesis inhibitory effect and / or a regulatory T cell inhibitory effect, thereby exhibiting antitumor activity. The compound of the present disclosure or a salt thereof has the ability to induce apoptosis. The compound of the present disclosure or a salt thereof can be administered intravenously and exhibits antitumor activity in a mouse model. Further, the compound of the present disclosure or a salt thereof has been confirmed to be safe in a mouse model.

[0022] As described above, the compound of the present disclosure or a salt thereof has improved water solubility and antitumor activity compared to the curcumin derivative of Patent Document 1, and further, its safety has also been confirmed. Therefore, the compound of the present disclosure or a salt thereof can be widely used as an antitumor agent containing these as active ingredients. The antitumor agent containing the compound of the present disclosure or a salt thereof as an active ingredient may be an antitumor agent for gastric cancer, an antitumor agent for colorectal cancer, an antitumor agent for pancreatic cancer, an antitumor agent for malignant mesothelioma, or an antitumor agent for cutaneous T cell lymphoma. Further, the antitumor agent may be an antitumor agent for gastric cancer, colorectal cancer, pancreatic cancer, malignant mesothelioma, and cutaneous T cell lymphoma, or an antitumor agent for pancreatic cancer, malignant mesothelioma, and cutaneous T cell lymphoma. Among them, the antitumor agent may be an antitumor agent for pancreatic cancer. Further, the compound of the present disclosure or a salt thereof can be used for various purposes as an active ingredient of an anti-inflammatory agent, an immunotherapy agent, a cardiac failure protective agent, or the like.

[0023] Note that the content of the active ingredient should be determined as an optimal amount in consideration of the patient's condition (general condition, disease state, presence or absence of complications), age, weight, and the like. The form of the drug is not particularly limited, and known forms such as oral preparations, injections, and inhalants can be used.

[0024] Next, a method for producing the compound of the present disclosure or a salt thereof will be described. The method for producing the compound of the present disclosure or a salt thereof is not particularly limited, but the following production method (the production method of the present disclosure) can be mentioned. In the production method of the present disclosure, the following compounds (1) to (3) are used. Compounds (1) and (3) are known and can be purchased from a reagent company. Compound (2) can be obtained by the method described in Patent Document 1.

[0025]

Chemical formula

[0026]

Chemical formula

[0027]

Chemical formula

[0028] The method for producing the compound of the present disclosure or a salt thereof includes a step S1 of acetylthiolating the hydroxyl group of compound (1), a step S2 of reacting the acetylthiolated compound (1) with compound (2) to obtain intermediate A, and a step S3 of reacting intermediate A with compound 3 to obtain the compound of the present disclosure. The reaction diagrams of each step are shown below.

[0029]

Chemical formula

[0030] Step S1 is not particularly limited as long as the hydroxyl group of compound (1) can be acetylthiolated. For example, the acetylthiolated compound (1) can be obtained by tosylating the hydroxyl group of compound (1) under basic conditions and then acetylthiolating the tosylated compound (1).

[0031] Tosylation is carried out by reacting compound (1) with a halide of p-toluenesulfonic acid under basic conditions. The solvent used for tosylation is not particularly limited as long as tosylation can proceed, and examples include ether. The method for obtaining basic conditions is not particularly limited, and for example, it can be obtained by dissolving potassium hydroxide in a solvent. Specific methods of tosylation are described in the examples below.

[0032] Acetylthiolation is carried out by reacting the tosylated compound (1) with thioacetic acid or its salt. The solvent used for acetylthiolation is not particularly limited, and examples include DMF. Specific methods of acetylthiolation are described in the examples below.

[0033] Step S2 is not particularly limited as long as the acetylthiolated compound (1) and compound (2) can react to obtain intermediate A. For example, after thiolating the acetylthiolated compound (1), intermediate A can be obtained by reacting the thiolated compound (1) with compound (2).

[0034] Thiolation proceeds by adding an alkoxide to an alcohol solution of the acetylthiolated compound (1). Specific methods of thiolation are described in the examples below.

[0035] The reaction between the thiolated compound (1) and compound (2) proceeds in the presence of a basic compound. The basic compound is not particularly limited, and examples include triethylamine. The solvent is not particularly limited, and examples include DMF. Specific methods of the reaction between the thiolated compound (1) and compound (2) are described in the examples below.

[0036] Step S3 is not particularly limited as long as it can react intermediate A with compound (3) to obtain the compound of the present disclosure. For example, it can be obtained by bonding the triple bond site of intermediate A and the azide site of compound (3) through a cyclization reaction to form a triazole skeleton.

[0037] The cyclization reaction proceeds in the presence of a copper catalyst. The copper catalyst is not particularly limited, and examples include copper 2-thiophenecarboxylate. The solvent is not particularly limited, and examples include a water-containing ether solvent. The specific method of the cyclization reaction is described in the examples below.

[0038] The production method of the present disclosure may include a step of converting the compound obtained in step S3 into a salt by a neutralization reaction. The method of converting the compound of the present disclosure into a salt by a neutralization reaction is not particularly limited, and a known method can be adopted. For example, the target salt can be obtained by dissolving the compound of the present disclosure in a basic solution containing a predetermined metal ion.

Examples

[0039] Based on the following examples, the present disclosure will be further described. However, the present disclosure is not limited thereto. Here, the compounds used in the experiments are shown in FIGS. 1 to 5.

[0040] [Synthesis of Compounds] GO-Y193, 197 to 200, and 206 were synthesized according to the following synthetic route. Also, GO-Y196 was synthesized according to a similar synthetic route.

[0041]

Chemical formula

[0042] [Synthesis of Compound 2] To a solution of 3,6,9,12-tetraoxapentadeca-14-in-1-ol (Compound 1, 3.87 g, 16.7 mmol) in diethyl ether (25 mL) was added potassium hydroxide (2.85 g, 20.0 mmol) at 0 °C. After stirring at 0 °C for 10 minutes, paratoluenesulfonyl chloride (3.82 g, 20.0 mmol) was added and stirring was continued at room temperature for 50 minutes. The reaction was stopped by adding half-saturated aqueous ammonium chloride solution (50 mL), and the mixture was extracted with ethyl acetate (160 mL + 80 mL). The combined organic layers were washed with saturated brine (80 mL) and dried over anhydrous sodium sulfate. The solution filtered through cotton was concentrated under reduced pressure to obtain a crude product (6.67 g) available for the next step.

[0043] To a solution of the crude product (crude tosylate) in N,N-dimethylformamide (20 mL) was added potassium S-acetate (2.30 g, 20.0 mmol) at 0 °C. After stirring at room temperature for 2.5 hours, the reaction was stopped by adding saturated aqueous sodium hydrogen carbonate solution (50 mL). The resulting mixture was filtered through cotton, and the filtrate was extracted with diethyl ether (4 × 100 mL). The combined organic layers were washed with saturated brine (100 mL) and dried over anhydrous sodium sulfate. The crude product (10.7 g) obtained by concentrating the solution filtered through cotton under reduced pressure was purified by flash column chromatography (silica gel 50 g, normal hexane / ethyl acetate 3:1 → ethyl acetate) to give S-3,6,9,12-tetraoxapentadeca-14-in-1-yl acetate (Compound 2, 3.54 g, two-step yield 73%) as an orange oily compound.

[0044] (Compound data of Compound 2) R f 0.63 (AcOEt); IR (neat) 3259, 2869, 2113, 1692, 1456, 1353, 1292, 1249, 1105, 1034, 954 cm -1 ; 11H NMR (400 MHz, CDCl3) δ 2.34 (s, 3H), 2.43 (t, J = 2.3 Hz, 1H), 3.09 (t, J = 6.5 Hz, 2H), 3.60 (t, J = 6.5 Hz, 2H), 3.63 - 3.72 (m, 12H), 4.21 (d, J = 2.3 Hz, 2H); 13 13C NMR (100 MHz, CDCl3) δ 28.8, 30.5, 58.4, 69.1, 69.7, 70.3, 70.4, 70.5, 70.6, 74.4, 74.5, 79.6, 195.5; HRMS (FAB) m / z [M + H] + calcd for C 13 H 23 O5S 291.1261; found 291.1238.

[0045] <Synthesis of Compound 3> To a methanol solution (10 mL) of Compound 2 (620 mg, 2.13 mmol) was added sodium methoxide (347 mg, 6.42 mmol) at 0 °C. After stirring at room temperature for 1.5 h, a cation exchange resin (Dowex 50Wx8) was added to neutralize the reaction mixture. After filtration through celite, the filtrate was concentrated under reduced pressure to obtain a crude product (734 mg) that could be used in the next step.

[0046] GO-Y030 (1.02 g, 2.14 mmol) and triethylamine (0.35 mL, 2.52 mmol) were dissolved in N,N-dimethylformamide (6.0 mL). A solution of the crude thiol in N,N-dimethylformamide (12 mL + 2 mL for washing) was slowly added at 23 °C. After stirring for 2 hours at the same temperature (23 °C), water (50 mL) was added. The resulting mixture was extracted with diethyl ether (3 × 100 mL). The combined organic layers were washed with saturated brine (150 mL) and then dried over anhydrous sodium sulfate. The solution filtered through a cotton plug was concentrated under reduced pressure to obtain a crude product (1.89 g), which was purified by flash column chromatography (50 g of silica gel, normal hexane / ethyl acetate 1:1) to give GO-Y030-SPEG4-alkyne (Compound 3, 776 mg, 50% yield) as a pale yellow oily compound.

[0047] Here, GO-YO30 used in the above reaction is (1E,4E)-1,5-bis-[3,5-bis(methoxymethoxy)phenyl]pentadiene-3-one. The synthesis method of GO-YO30 is described in Patent Document 1.

[0048] (Compound data of Compound 3) R f 0.15 (n-Hexane / AcOEt 1:1); IR (neat) 3279, 2901, 2827, 2114, 1688, 1663, 1592, 1454, 1401, 1332, 1281, 1248, 1215, 1146, 1185, 1033, 965 cm -1 ; 11H NMR (400 MHz, acetone-d6) δ 2.56 (t, J = 6.5 Hz, 2H), 2.92 (t, J = 2.5 Hz, 1H), 3.22 (dd, J = 7.3, 16.4 Hz, 1H), 3.30 (dd, J = 7.3, 16.4 Hz, 1H), 3.41 (s, 6H), 3.43 (s, 6H), 3.50 - 3.61 (m, 14H), 4.54 (d, J = 2.5 Hz, 2H), 4.53 (t, J = 7.3 Hz, 1H), 5.16 (s, 4H), 5.22 (s, 4H), 6.59 (t, J = 2.2 Hz, 1H), 6.76 (t, J = 2.3 Hz, 1H), 6.78 (d, J = 2.3 Hz, 2H), 6.82 (d, J = 15.9 Hz, 1H), 7.00 (d, J = 2.3 Hz, 2H), 7.57 (d, J = 15.9 Hz, 1H); 13 13C NMR (150 MHz, acetone-d6) δ 31.3, 45.6, 47.5, 56.1, 56.2, 58.5, 69.8, 70.9, 71.0, 71.2, 71.6, 75.7, 81.0, 95.1, 95.2, 104.1, 107.7, 110.20, 110.23, 127.8, 137.7, 143.1, 145.7, 159.3, 159.6, 197.0; HRMS (FAB) m / z [M + H] + calcd for C 36 H 51 O 13 S 723.3045; found 723.3062.

[0049] <Synthesis of GO - Y193> Compound 3 (214 mg, 296 μmol) and azide (358 mg, 1.75 mmol) were dissolved in aqueous tetrahydrofuran (8.1 mL, tetrahydrofuran:water = 100:1). After adding copper(II) 2-thiophenecarboxylate (4.1 mg, 21.5 μmol), the mixture was stirred at room temperature for 2 hours. The residue obtained by concentrating the reaction solution under reduced pressure was purified by flash column chromatography (2.4 g of silica gel, chloroform / methanol 19:1 → 9:1) to give GO-Y193 (62.0 mg, yield 23%) as a colorless amorphous solid.

[0050] (Compound data of GO-Y193) R f 0.67 (CHCl3 / MeOH 4:1); IR (neat) 3398, 1688, 1660, 1593, 1455, 1401, 1281, 1215, 1146, 1085, 1033, 924 cm -1 ; 11H NMR (600 MHz, CD3OD) δ 2.55 (t, J = 6.5 Hz, 2H), 3.18 (dd, J = 7.0, 16.1 Hz, 1H), 3.24 (dd, J = 7.9, 16.1 Hz, 1H), 3.41 (s, 6H), 3.44 (s, 6H), 3.50 - 3.64 (m, 17H), 3.72 (dd, J = 5.5, 12.2 Hz, 1H), 3.88 (dd, J = 2.0, 12.2 Hz, 1H), 3.90 (t, J = 9.0 Hz, 1H), 4.47 (dd, J = 7.0, 7.9 Hz, 1H), 4.62 (s, 2H), 5.12 (d, J = 2.2 Hz, 2H), 5.13 (d, J = 2.2 Hz, 2H), 5.18 (s, 4H), 5.60 (d, J = 9.1 Hz, 1H), 6.59 (t, J = 2.2 Hz, 1H), 6.75 (d, J = 16.1 Hz, 1H), 6.76 (t, J = 2.2 Hz, 1H), 6.76 (d, J = 2.2 Hz, 2H), 6.92 (d, J = 2.2 Hz, 2H), 7.48 (d, J = 16.1 Hz, 1H), 8.15 (s, 1H); 1313C NMR (150 MHz, CD3OD) δ 31.6 (CH2), 46.4 (CH), 48.0 (CH2), 56.3 (CH3), 56.4 (CH3), 62.4 (CH2), 64.9 (CH2), 70.8 (CH2), 70.9 (CH), 71.3 (CH2), 71.51 (CH2), 71.53 (CH2), 71.55 (CH2), 72.0 (CH2), 74.0 (CH), 78.5 (CH), 81.1 (CH), 89.5 (CH), 95.5 (CH2), 95.6 (CH2), 104.7 (CH), 108.2 (CH), 110.6 (CH), 110.7 (CH), 124.3 (CH), 127.8 (CH), 137.9 (C), 144.6 (CH), 145.8 (C), 146.1 (C), 159.7 (C), 160.0 (C), 199.5 (C); HRMS (FAB) m / z [M + H] + calcd for C 42 H 62 N3O 18 S 928.37442; found 928.3762.

[0051] <Synthesis of GO-Y197> Compound 3 (372 mg, 515 μmol), azide (164 mg, 799 μmol), aqueous tetrahydrofuran (10 mL, tetrahydrofuran:water = 100:1), copper 2-thiophenecarboxylate (5.2 mg, 27.3 μmol) were used. The residue obtained by concentrating the reaction solution under reduced pressure was purified by flash column chromatography (10 g of silica gel, ethyl acetate → ethyl acetate / methanol 9:1) to give GO-Y197 (307 mg, 64% yield) as a colorless amorphous solid.

[0052] (Compound data of GO-Y197) R f 0.10 (AcOEt / MeOH 9:1); IR (neat) 3399, 2903, 1662, 1593, 1455, 1401, 1281, 1215, 1146, 1085, 1033, 924 cm-1 ; 1 1H NMR (600 MHz, CD3OD) δ 2.53 (t, J = 6.2 Hz, 2H), 3.18 (dd, J = 6.9, 16.0 Hz, 1H), 3.23 (dd, J = 7.9, 16.0 Hz, 1H), 3.39 (s, 6H), 3.42 (s, 6H), 3.50 - 3.62 (m, 17H), 3.71 (dd, J = 5.1, 12.2 Hz, 1H), 3.87 (d, J = 12.2 Hz, 1H), 3.91 (t, J = 9.1 Hz, 1H), 4.46 (dd, J = 6.9, 7.9 Hz, 1H), 4.61 (s, 2H), 5.11 (d, J = 6.8 Hz, 2H), 5.15 (d, J = 6.8 Hz, 2H), 5.16 (s, 4H), 5.61 (d, J = 9.1 Hz, 1H), 6.58 (t, J = 2.0 Hz, 1H), 6.73 (d, J = 16.1 Hz, 1H), 6.75 (t, J = 2.1 Hz, 1H), 6.75 (d, J = 2.0 Hz, 2H), 6.91 (d, J = 2.0 Hz, 2H), 7.46 (d, J = 16.1 Hz, 1H), 8.19 (s, 1H); 1313C NMR (150 MHz, CD3OD) δ 31.6 (CH2), 46.3 (CH), 48.0 (CH2), 56.38 (CH3), 56.43 (CH3), 62.4 (CH2), 65.0 (CH2), 70.78 (CH2), 70.82 (CH), 71.2 (CH2), 71.43 (CH2), 71.46 (CH2), 71.49 (CH2), 71.9 (CH2), 74.0 (CH), 78.4 (CH), 81.1 (CH), 89.6 (CH), 95.5 (CH2), 95.6 (CH2), 104.7 (CH), 108.2 (CH), 110.6 (CH), 110.7 (CH), 124.8 (CH), 127.8 (CH), 137.9 (C), 144.5 (CH), 145.8 (C), 159.7 (C), 159.9 (C), 199.4 (C); HRMS (FAB) m / z [M + H] + calcd for C 42 H 62 N3O 18 S 928.3744; found 928.3742.

[0053] <Synthesis of GO-Y198> Compound 3 (247 mg, 342 μmol), azide (127 mg, 340 μmol), aqueous tetrahydrofuran (7.0 mL, tetrahydrofuran:water = 100:1), copper 2-thiophenecarboxylate (5.2 mg, 27.3 μmol) were used. The residue obtained by concentrating the reaction solution under reduced pressure was purified by flash column chromatography (2.3 g silica gel, chloroform / methanol 19:1 → 9:1) to give GO-Y198 (102 mg, 32% yield) as a colorless amorphous solid.

[0054] (Compound data of GO-Y198) R f0.13 (AcOEt / MeOH 9:1); IR (neat) 3399, 2903, 1685, 1654, 1593, 1456, 1400, 1332, 1281, 1215, 1146, 1084, 1033, 924 cm -1 ; 1 1H NMR (600 MHz, CD3OD) δ 2.53 (t, J = 6.5 Hz, 2H), 3.16 (dd, J = 7.0, 15.8 Hz, 1H), 3.20 (dd, J = 7.9, 15.8 Hz, 1H), 3.39 (s, 6H), 3.43 (s, 6H), 3.49 - 3.52 (m, 4H), 3.55 - 3.63 (m, 11H), 3.72 - 3.82 (m, 3H), 4.07 (dd, J = 3.3, 8.7 Hz, 1H), 4.45 (dd, J = 6.9, 7.9 Hz, 1H), 4.68 (t, J = 3.3 Hz, 1H), 4.80 (s, 2H), 5.11 (d, J = 6.8 Hz, 2H), 5.14 (d, J = 6.8 Hz, 2H), 5.16 (s, 4H), 6.02 (d, J = 2.6 Hz, 1H), 6.57 (t, J = 2.1 Hz, 1H), 6.72 (d, J = 16.1 Hz, 1H), 6.74 (t, J = 2.1 Hz, 1H), 6.74 (d, J = 2.0 Hz, 2H), 6.90 (d, J = 2.0 Hz, 2H), 7.46 (d, J = 16.1 Hz, 1H), 8.12 (s, 1H); 1313C NMR (150 MHz, CD3OD) δ 31.6 (CH2), 46.3 (CH), 48.0 (CH2), 56.37 (CH3), 56.42 (CH3), 62.5 (CH2), 64.9 (CH2), 68.5 (CH), 70.1 (CH), 70.8 (CH2), 71.2 (CH2), 71.45 (CH2), 71.48 (CH2), 71.51 (CH2), 71.9 (CH2), 72.5 (CH), 78.5 (CH), 88.3 (CH), 95.5 (CH2), 95.6 (CH2), 104.7 (CH), 108.2 (CH), 110.6 (CH), 110.7 (CH), 125.0 (CH), 127.8 (CH), 137.9 (C), 144.5 (CH), 145.8 (C), 146.3 (C), 159.7 (C), 159.9 (C), 199.3 (C); HRMS (FAB) m / z [M + H] + calcd for C 42 H 62 N3O 18 S 928.3744; found 928.3752.

[0055] <Synthesis of GO-Y199> Compound 3 (214 mg, 296 μmol), azide (109 mg, 296 μmol), aqueous tetrahydrofuran (6.0 mL, tetrahydrofuran:water = 100:1), copper(II) 2-thiophenecarboxylate (6.2 mg, 32.5 μmol) were used. The residue obtained by concentrating the reaction solution under reduced pressure was purified by flash column chromatography (twice: 3 g of silica gel, chloroform / methanol 9:1 → 4:1, 2.3 g of silica gel, chloroform / methanol 9:1 → 17:3) to give GO-Y199 (199 mg, 62% yield) as a colorless amorphous solid.

[0056] (Compound data of GO-Y199) R f0.20 (CHCl3 / MeOH 4:1); IR (neat) 3389, 1660, 1593, 1455, 1440, 1401, 1333, 1281, 1241, 1215, 1146, 1084, 1034, 924 cm -1 ; 1 1H NMR (600 MHz, CD3OD) δ 2.55 (t, J = 6.6 Hz, 2H), 3.18 (dd, J = 7.1, 15.9 Hz, 1H), 3.23 (dd, J = 7.9, 15.9 Hz, 1H), 3.41 (s, 6H), 3.45 (s, 6H), 3.47-3.64 (m, 17H), 3.74-3.83 (m, 6H), 3.89 (m, 2H), 3.97 (t, J = 9.1 Hz, 1H), 4.41 (d, J = 7.8 Hz, 1H), 4.47 (dd J = 6.6, 7.9 Hz, 1H), 4.62 (s, 2H), 5.12 (d, J = 6.9 Hz, 2H), 5.14 (d, J = 6.9 Hz, 2H), 5.18 (s, 4H), 5.63 (d, J = 9.1 Hz, 1H), 6.58 (t, J = 2.2 Hz, 1H), 6.74 (d, J = 16.2 Hz, 1H), 6.76 (t, J = 2.1 Hz, 1H), 6.76 (d, J = 2.2 Hz, 2H), 6.92 (d, J = 2.1 Hz, 2H), 7.48 (d, J = 16.2 Hz, 1H), 8.16 (s, 1H); 1313C NMR (150 MHz, CD3OD) δ 31.6 (CH2), 46.4 (CH), 48.1 (CH2), 56.3 (CH3), 56.4 (CH3), 61.6 (CH2), 62.5 (CH2), 65.0 (CH2), 70.3 (CH), 70.8 (CH2), 71.3 (CH2), 71.51 (CH2), 71.56 (CH2), 71.57 (CH2), 71.59 (CH2), 72.0 (CH2), 72.5 (CH), 73.7 (CH), 74.9 (CH), 76.9 (CH), 77.1 (CH), 79.6 (CH), 79.8 (CH), 89.3 (CH), 95.56 (CH2), 95.62 (CH2), 104.8 (CH), 105.1 (CH), 108.2 (CH), 110.65 (CH), 110.72 (CH), 124.3 (CH), 127.8 (CH), 137.9 (C), 144.6 (CH), 145.8 (C), 146.1 (C), 159.8 (C), 160.0 (C), 199.6 (C); HRMS (FAB) m / z [M + H] + calcd for C 48 H 72 N3O 23 S 1090.4272; found 1090.4265.

[0057] <Synthesis of GO-Y200> Compound 3 (226 mg, 313 μmol), azide (160 mg, 436 μmol), aqueous tetrahydrofuran (10 mL, tetrahydrofuran:water = 100:1), and copper(II) 2-thiophenecarboxylate (3.3 mg, 17.3 μmol) were used. The residue obtained by concentrating the reaction solution under reduced pressure was purified by flash column chromatography (5.5 g of silica gel, chloroform / methanol 9:1 → 17:3) to give GO-Y200 (133 mg, 35% yield) as a colorless amorphous solid.

[0058] (Compound data of GO-Y200) R f0.08 (CHCl3 / MeOH 8:1); IR (neat) 3386, 2904, 1684, 1661, 1594, 1541, 1506, 1456, 1400, 1334, 1281, 1216, 1146, 1084, 1033, 968 cm -1 ; 1 1H NMR (600 MHz, CD3OD) δ 2.56 (t, J = 6.5 Hz, 2H), 3.18 (dd, J = 6.9, 16.0 Hz, 1H), 3.24 (dd, J = 7.9, 16.0 Hz, 1H), 3.42 (s, 6H), 3.46 (s, 6H), 3.48 (m, 1H), 3.53 - 3.56 (m, 4H), 3.59 - 3.70 (m, 12H), 3.76 (t, J = 9.2 Hz, 1H), 3.82 - 3.90 (m, 4H), 3.96 (t, J = 9.2 Hz, 1H), 4.48 (dd J = 6.9, 7.9 Hz, 1H), 4.63 (s, 2H), 5.13 (d, J = 6.8 Hz, 2H), 5.15 (d, J = 6.8 Hz, 2H), 5.19 (s, 4H), 5.24 (d, J = 3.9 Hz, 1H), 5.63 (d, J = 9.2 Hz, 1H), 6.59 (t, J = 2.2 Hz, 1H), 6.75 (d, J = 16.3 Hz, 1H), 6.77 (t, J = 2.2 Hz, 1H), 6.77 (d, J = 2.2 Hz, 2H), 6.94 (d, J = 2.2 Hz, 2H), 7.49 (d, J = 16.3 Hz, 1H), 8.17 (s, 1H); 1313C NMR (150 MHz, CD3OD) δ 31.6 (CH2), 46.4 (CH), 48.1 (CH2), 56.3 (CH3), 56.4 (CH3), 61.9 (CH2), 62.8 (CH2), 65.0 (CH2), 70.8 (CH-), 71.3 (CH2), 71.53 (CH), 71.57 (CH2), 71.58 (CH2), 71.60 (CH2), 72.1 (CH2), 73.6 (CH), 74.2 (CH), 74.9 (CH), 75.1 (CH), 78.2 (CH), 79.7 (CH), 80.4 (CH), 89.4 (CH), 95.57 (CH2), 95.63 (CH2), 103.0 (CH), 104.8 (CH), 108.2 (CH), 110.66 (CH), 110.72 (CH), 124.3 (CH), 127.8 (CH), 138.0 (C), 144.6 (CH), 145.8 (C), 146.1 (C), 159.8 (C), 160.0 (C), 199.6 (C); HRMS (FAB) m / z [M + H] + calcd for C 48 H 72 N3O 23 S 1090.4272; found 1090.4303.

[0059] <Synthesis of GO-Y206> Compound 3 (148 mg, 205 μmol), azide (α:β = 1:0.7, 116 mg, 613 μmol), aqueous tetrahydrofuran (4.0 mL, tetrahydrofuran:water = 100:1), and copper(II) 2-thiophenecarboxylate (8.8 mg, 46.2 μmol) were used. The residue obtained by concentrating the reaction solution under reduced pressure was purified by flash column chromatography (silica gel 5.0 g, chloroform / methanol 1:0 → 100:3 → 10:1) to give GO-Y206 (113 mg, 61% yield) as a colorless oily substance. Note that Figure 5 shows only the compound with β-deoxyglucose attached, but GO-Y206 also includes the compound with α-deoxyglucose attached.

[0060] (Compound data of GO-Y206) R f 0.13 (AcOEt / MeOH 9:1); IR (neat) 3416, 2902, 1687, 1661, 1593, 1454, 1400, 1332, 1281, 1215, 1146, 1084, 1033, 924 cm -1 ; 1 1H NMR (600 MHz, CD3OD) δ 2.19 (q, J = 11.8 Hz, 1H), 2.43 (ddd, J = 2.0, 4.8, 11.8 Hz, 1H), 2.52 (t, J = 6.7 Hz, 2H), 3.18 (dd, J = 7.0, 16.0 Hz, 1H), 3.23 (dd, J = 7.4, 16.0 Hz, 1H), 3.39 (t, J = 9.3 Hz, 1H), 3.40 (s, 6H), 3.43 (s, 6H), 3.49 - 3.63 (m, 15H), 3.72 (dd, J = 5.4, 12.0 Hz, 1H), 3.79 (m, 1H), 3.89 (dd, J = 2.3, 12.0 Hz, 1H), 4.46 (t, J = 7.4 Hz, 1H), 4.60 (s, 2H), 5.11 (d, J = 7.0 Hz, 2H), 5.13 (d, J = 7.0 Hz, 2H), 5.17 (s, 4H), 5.90 (dd, J = 2.0, 11.8 Hz, 1H), 6.58 (t, J = 2.2 Hz, 1H), 6.74 (d, J = 16.1 Hz, 1H), 6.75 (m, 3H), 6.91 (d, J = 2.2 Hz, 2H), 7.48 (d, J = 16.1 Hz, 1H), 8.15 (s, 1H); 1313C NMR (150 MHz, CD3OD) δ 31.6 (CH2), 39.4 (CH2), 46.3 (CH), 48.0 (CH2), 56.8 (CH3), 56.43 (CH3), 62.5 (CH2), 64.9 (CH2), 70.8 (CH2), 71.2 (CH2), 71.45 (CH2), 71.49 (CH2), 71.51 (CH2), 71.53 (CH2), 71.9 (CH2), 72.3 (CH), 72.4 (CH), 81.0 (CH), 85.6 (CH), 95.5 (CH2), 95.6 (CH2), 104.7 (CH), 108.2 (CH), 110.6 (CH), 110.7 (CH), 123.8 (CH), 127.8 (CH), 137.9 (C), 144.5 (CH), 145.8 (C), 146.1 (C), 159.7 (C), 159.9 (C), 199.3 (C); HRMS (FAB) m / z [M + H] + calcd for C 42 H 62 N3O 17 S 912.3794; found 912.3803.

[0061] [Turbidity measurement] The turbidity of various compounds was measured. The method is as follows. The compound was first dissolved in DMSO (dimethyl sulfoxide), and then dissolved in distilled water or DMEM medium (Dulbecco's modified Eagle's medium) containing 10% fetal bovine serum to a final concentration of 50 μM. Curcumin was turbid, and GO-Y199 was transparent. The solubility was measured with a turbidimeter (REX, WZB-170 Portable Turbidimeter). The turbidity of the compound was corrected for concentration. The results are shown in Table 1.

[0062]

Table 1

[0063] From Table 1, it was confirmed that the turbidity of GO-Y196 to 200 was 0 or very small, and the water solubility was improved compared to GO-Y030 and curcumin.

[0064] Subsequently, the final concentrations of GO-Y206 and curcumin were adjusted to 1.25 μM and 5.00 μM, respectively, and the turbidity of each was measured. The results are shown in Table 2.

[0065]

Table 2

[0066] The turbidity of GO-Y206 with a final concentration of 1.25 μM was 0.18 NTU, and the turbidity of GO-Y206 with a final concentration of 5.00 μM was 1.82 NTU. On the other hand, the turbidity of curcumin with a final concentration of 1.25 μM was 4.96 NTU, and the turbidity of curcumin with a final concentration of 5.00 μM was 29.0 NTU. When the NTU was divided by the concentration and the average of the obtained values was calculated, GO-Y206 was 0.254 NTU / μM, while curcumin was 4.884 NTU / μM. Therefore, the turbidity of GO-Y206 was improved to 5% of that of curcumin.

[0067] [Cell growth inhibitory activity and turbidity] The cell growth inhibitory activity against the colon cancer cell line HCT116 was measured. As an index of cell growth inhibitory activity, the concentration (IC 50 ) that inhibits cell growth by 50% compared to the control was used. The method is as follows. HCT116 was seeded in a 6-well plate at 5 x 10 4 per well, and after 24 hours, compounds at various concentrations were added. After further culturing for 72 hours, the number of cells was counted. The control used the same amount of DMSO as the amount added according to the highest concentration of the compound. The number of cells at each concentration relative to the control was shown as a percentage. The results are shown in Table 2. Also, in Table 3, the turbidity of the compounds was described. Furthermore, as represented by Figure 6, the measurement results of the cell growth inhibitory activity of GO-Y190, GO-Y193, GO-Y196, and GO-Y197 were shown.

[0068]

Table 3

[0069] From Table 3, GO-Y190 to 194, 196 to 200 had higher cell growth inhibitory activities compared to GO-Y030 and curcumin. Among them, GO-Y190, 191, 198 to 200 had particularly high cell growth inhibitory activities. Furthermore, among them, GO-Y198 to 200 had excellent water solubility. Since GO-Y199 was the one that could be synthesized with the best yield among these, subsequent experiments were conducted focusing on GO-Y199.

[0070] [Cell growth inhibitory activities against various cancer cell lines] The cell growth inhibitory activities (IC 50 ) of curcumin, GO-Y030, GO-Y193, and GO-Y199 against various cancer cell lines (gastric cancer cell lines: GCIY, Kato III, SH-10-TC, colorectal cancer cell lines: DLD-1, HT29, HCT-116, breast cancer cell lines: HCC38, HCC70, HCC1395) were measured. The method was as described above. The results are shown in Table 4. Also, in Table 5, the IC 50 of GO-Y199 and that of curcumin·GO-Y030 were compared respectively. Furthermore, the results of the cell growth inhibitory activity of GO-Y199 against various cancer cell lines are shown in Figure 7.

[0071]

Table 4

[0072]

Table 5

[0073] From Table 4, both GO-Y193 and GO-Y199 had high cell growth inhibitory activities against all cancer cell lines. Also, from Table 5, GO-Y199 had antitumor activities 33.3 - 71.4 times that of curcumin and antitumor activities 0.8 - 4.2 times that of GO-Y030. From these results, it can be said that GO-Y199 has higher antitumor activities than curcumin and GO-Y030.

[0074] Cell growth inhibitory activities (IC 50 ) of curcumin, GO-Y199, and GO-Y206 against colon cancer cell lines DLD-1, HCT116, gastric cancer cell lines KATO III, H-111-TC were measured. The method was as described above. The results are shown in Table 6. Also, the results of the cell growth inhibitory activities of these compounds against various cancer cell lines are shown in Figure 8.

[0075]

Table 6

[0076] From Table 6, both GO-Y199 and GO-Y206 had high cell growth inhibitory activities against all colon cancer cell lines compared to curcumin. Also, the effect of GO-Y206 was equivalent to that of GO-Y199.

[0077] Cell growth inhibitory activities (IC 50 ) of curcumin, GO-Y022, GO-Y030, GO-Y199, and GO-Y200 against pancreatic cancer cell lines ASPC-1, Panc-1 were measured. The method was as described above. The results are shown in Table 7. Also, the results of the cell growth inhibitory activities of these compounds against various cancer cell lines are shown in Figure 9.

[0078]

Table 7

[0079] From Table 7, GO-Y199 and GO-Y200 had higher cell growth inhibitory activities against all pancreatic cancer cell lines compared to curcumin. Also, GO-Y199 and GO-Y200 had higher effects compared to GO-Y022.

[0080] The cell growth inhibitory activities (IC 50 ) of curcumin, GO-Y193, GO-Y197, GO-Y198, GO-Y199, and GO-Y200 against malignant pleural mesothelioma cell lines NCI-H226 and MSTO211H were measured. The method was as described above. The results are shown in Table 8 and Figure 10. For comparison, the IC 50 of the cytotoxic anticancer drug cisplatin (CDDP) was also included.

[0081]

Table 8

[0082] From Table 8, GO-Y193, GO-Y197, GO-Y198, GO-Y199, and GO-Y200 had higher cell growth inhibitory activities against all malignant pleural mesothelioma cell lines compared to curcumin.

[0083] The cell growth inhibitory activities (IC 50 ) of curcumin, GO-Y022, GO-Y030, GO-Y193, GO-Y197, and GO-Y199 against malignant melanoma cell line G361 were measured. The method was as described above. The results are shown in Table 9. Also, the results of the cell growth inhibitory activities of GO-Y193, GO-Y197, and GO-Y199 against malignant melanoma cell lines are shown, represented by Figure 11.

[0084]

Table 9

[0085] From Table 9, GO-Y193, GO-Y197, and GO-Y199 had higher cell growth inhibitory activity against melanoma cell lines compared to curcumin. Also, GO-Y193, GO-Y197, and GO-Y199 had a higher effect than GO-Y022 and GO-Y030.

[0086] The cell growth inhibitory activities (IC 50 ) of curcumin, GO-Y022, GO-Y030, GO-Y193, GO-Y197, GO-Y198, GO-Y199, and GO-Y200 against cutaneous T-cell lymphoma cell lines HH and HUT78 were measured. The method was as described above. The results are shown in Table 10. Also, the results of the cell growth inhibitory activities of these compounds against various cancer cell lines are shown in Figure 12.

[0087]

Table 10

[0088] From Table 10, GO-Y193, GO-Y197, GO-Y198, GO-Y199, and GO-Y200 had higher cell growth inhibitory activity against all cutaneous T-cell lymphoma cell lines compared to curcumin. Also, GO-Y193, GO-Y197, GO-Y198, GO-Y199, and GO-Y200 had a higher effect than GO-Y022.

[0089] [Cytotoxic effect] The cytotoxic effect of GO-Y199 was investigated. The method was as follows. Each cell line was seeded at 5x10 4 in a 6-well plate, and after 24 hours, 2 μM and 5 μM of GO-Y199 were added. The cell count was performed after 48 hours and 72 hours. The results are shown in Figure 13.

[0090] As shown in Figure 13, a cytotoxic effect of GO-Y199 was observed against both cancer cell lines, HCT116 and Kato III.

[0091] [Effect on NF-kB] The effect of GO-Y199 on NF-kB (p65) was examined. The method was as follows. The HCT116 cell line was seeded at 3x10 6 cells in a 10-cm plate. After 24 hours, 2 μM of GO-Y199 was added. After 24 hours, the cells were collected, embedded in iPGell (GenoStaff), and paraffin blocks were prepared after formalin fixation. Immunohistochemistry was performed with an anti-NF-kB (p65) antibody. Also, 2 μM and 5 μM of GO-Y199 were added, and after 24 hours, the lysates were collected, and the expression level of NF-kB (p65) was analyzed using an ELISA plate (NF-kB p65 (pS536) SimpleStep ELISA kit, Abcam). The protein amount was corrected with the absorbance at A280 nm. The results are shown in Figures 14 and 15. The upper part of Figure 14 is a photograph of the control group and the GO-Y199 (2 μM)-added group after 24 hours. The lower part of Figure 14 is the result of the t-test for these groups after 24 hours. Figure 15 is the comparison result of the NF-kB inhibitory effect of the GO-Y199 (2 μM, 5 μM)-added groups against the control group.

[0092] As shown in Figure 14, the proportion of NF-kB (p65)-expressing cells in the control group was 96.8 ± 1.1%, while that in the GO-Y199-added group was 69.4 ± 12.4%. Also, as shown in Figure 15, the relative expression level of NF-kB (p65) in the control group was 5.27 ± 0.95, while those in the GO-Y199-added groups (2 μM, 5 μM) were 0 and 0, respectively. From this, it was confirmed that GO-Y199 has an NF-kB inhibitory effect.

[0093] [Effect on pSTAT3] The effect of GO-Y199 on pSTAT3 was examined. The method was as described above. Immunohistochemistry was performed with an anti-pSTAT3 antibody. The results are shown in Figure 16. The upper part of Figure 16 is a photograph of the control group and the GO-Y199 (2 μM)-added group after 24 hours. The lower part of Figure 16 is the result of the t-test for these groups after 24 hours.

[0094] As shown in Fig. 16, the relative expression level of pSTAT3 in the control group was 10.2 ± 1.6%, while that in the GO-Y199 (2 μM) addition group was 0.3 ± 0.5%. Thus, since the expression level of pSTAT3 was significantly decreased by the addition of GO-Y199, it was confirmed that GO-Y199 inhibits the expression of pSTAT3.

[0095] [Effect on β-catenin] The effect of GO-Y199 on β-catenin was examined. The method was as described above. Immunohistochemistry was performed with an anti-β-catenin antibody. Protein quantification was performed using an ELISA plate (proteintech). The results are shown in Figs. 17 and 18. Fig. 17 is a photograph of the control group and the GO-Y199 (2 μM) addition group after 24 hours. The numerical values in Fig. 17 are the results of a t-test of β-catenin-inactivated cells. Fig. 18 is a comparison result of the β-catenin inhibitory effect of the GO-Y199 (2 μM, 5 μM) addition groups with respect to the control group.

[0096] As shown in Fig. 18, the relative expression level of β-catenin was 11.5 ± 2.3 in the control group, 2.8 ± 0.8 in the GO-Y199 (2 μM) addition group, and 3.9 ± 1.1 in the GO-Y199 (5 μM) addition group. The expression level of β-catenin was significantly decreased by the addition of GO-Y199. Therefore, it was confirmed that GO-Y199 has a β-catenin inhibitory effect.

[0097] [Effect on fatty acid synthase] The effect of GO-Y199 on fatty acid synthase was examined. The method was as described above. Protein quantification was performed using a Human Fatty acid synthase ELISA Kit (MyBioSource). The results are shown in Fig. 19. Fig. 19 is a comparison result of the fatty acid synthase inhibitory effect of the GO-Y199 (4 μM) addition group with respect to the control group.

[0098] As shown in Fig. 19, it was confirmed that GO-Y199 has a significant fatty acid synthase inhibitory effect.

[0099] [Effect on caspase 3] The effect of GO-Y199 on caspase 3 was examined. The method was as described above. Immunohistochemistry was performed with an anti-caspase 3 antibody. Quantification of apoptosis-related proteins was performed using M30 Apotosense ELISA (PEVIVA). The results are shown in Figs. 20 and 21. The upper part of Fig. 20 is a photograph of the control group and the GO-Y199 (2 μM)-added group after 24 hours. The lower part of Fig. 20 is the proportion of caspase 3-expressing cells after 24 hours. Fig. 21 shows the induction of apoptosis-related proteins in the GO-Y199 (2 μM, 5 μM)-added groups relative to the control group.

[0100] As shown in Fig. 20, the proportion of caspase 3-expressing cells in the control group was 1.0 ± 0.1%, while in the GO-Y199 (2 μM)-added group, it increased significantly to 57.4 ± 2.6%. Also, as shown in Fig. 21, while the control group was 360 U / L, the GO-Y199 (2 μM)-added group was 1,830 U / L, and the GO-Y199 (5 μM)-added group was 2,050 U / L, indicating the induction of apoptosis-related proteins by the addition of GO-Y199. Therefore, it was confirmed that GO-Y199 activates caspase 3 and induces apoptosis.

[0101] [Angiogenesis inhibitory activity] The angiogenesis inhibitory activities of curcumin, GO-Y022, GO-Y030, GO-Y193, GO-Y197, GO-Y198, GO-Y199, and GO-Y200 against the angiogenesis inhibitor-resistant vascular endothelial cells HUVEC-R were examined. HUVEC-R was cultured in the EGMTM-2 Bullet Kit (Takara Bio Inc., Otsu, Japan) in the presence of each compound, and the cell number was measured after 72 hours. Also, from the results, IC 50It was determined. GO-Y030 is known to have angiogenesis inhibitory activity, and a comparative study was conducted with it. The results are shown in Table 11 and Figure 22.

[0102]

Table 11

[0103] From Table 11, GO-Y193, GO-Y197, GO-Y198, GO-Y199, and GO-Y200 had higher angiogenesis inhibitory activity compared to curcumin. Also, GGO-Y193, GO-Y197, GO-Y198, GO-Y199, and GO-Y200 had a higher effect than GO-Y022.

[0104] [Regulatory T cell inhibitory activity] CD4 + CD62L hi Using the T Cell Isolation Kit (Miltenyi Biotec), naive CD4 + T cells were collected from the spleens of mice. 0.5×10 6 cells / mL of the collected T cells were added to RPMI 1640 medium supplemented with penicillin / streptomycin (5,000 units / mL), 10% fetal bovine serum, and 50 μM 2-mercaptoethanol. Subsequently, 1 μg / mL of anti-CD3 antibody (eBioscience) was added to the medium, and further 1 μg / ml of anti-CD28 antibody (eBioscience) was added. Then, the medium was cultured at 37°C for 3 days. Next, TGF-β1 (2 ng / mL) was added to the medium and cultured for 24 hours. Then, the cells were fixed using the FOXP3 Staining Buffer Kit (eBioscience), the cell membrane was treated, and nuclear FOXP3 was stained. The stained cells were measured using a BD LSRFortessaTM (BD Bioscience) flow cytometer, and the obtained data were analyzed with FlowJo (Tree-Star version). The results are shown in Figure 23.

[0105] The induction rate of FOXP3-positive regulatory T cells as the control group was 12.8%, but the induction rate of regulatory T cells with the addition of 0.3 μM GO-Y199 was suppressed to 2.22%.

[0106] [In vivo antitumor effect] The in vivo antitumor effect of GO-Y199 was investigated. The method was as follows. HCT116 was transplanted subcutaneously into nude mice. After tumor formation, a DMSO solution of 100 mM GO-Y199 was dissolved in 100 μL of PBS and injected once via the tail vein of the mice (equivalent to 1 mg of GO-Y199). As a control group, an equivalent concentration of DMSO-PBS solution was injected once via the tail vein. Four days after administration, the maximum diameter of the tumor was measured. The results are shown in Figure 24.

[0107] Figure 24 shows the degree of relative tumor growth in the control group (left) and the GO-Y199 intravenous injection group (right). The tumor growth in the control group was 113.7 ± 12.4% before administration, while in the GO-Y199 intravenous injection group, it was 95.2 ± 20.0% before administration, indicating that the addition of GO-Y199 significantly suppressed tumor growth. Therefore, it was confirmed that GO-Y199 exhibits antitumor activity in vivo.

[0108] Using an animal model, the antitumor effect of GO-Y199 on malignant mesothelioma cells was investigated. The method was as follows. Each of 2.8×10 6 MSTO-211H cells were transplanted into the abdominal cavity of 5 nude mice. From Day 5, 43 μL of a DMSO solution of 100 mM GO-Y199 was diluted to 500 μL with PBS and administered intraperitoneally to the mice (equivalent to 5 mg of GO-Y199). As a control group, an equivalent concentration of DMSO-PBS solution was administered intraperitoneally to 4 nude mice transplanted with MSTO-211H cells into the abdominal cavity in the same manner. The administration was performed 5 times at 5-day intervals. After 30 days, the mice were sacrificed, dissected, and analyzed. The results are shown in Figure 25.

[0109] In one of the GO-Y199-administered group, tumor formation was observed at the puncture site where MSTO-211H cells were intraperitoneally administered. This was not observed in the control group. Minute peritoneal dissemination lesions were observed in 4 (80%) of the GO-Y199-administered group and 3 (75%) of the control group. On the other hand, one (20%) of the GO-Y199-administered group had 1 seeded nodule exceeding 5 mm in maximum diameter, while a total of 6 were observed in 4 (100%) of the control group. Also, the size (maximum diameter) was 5 mm in the GO-Y199-administered group, while it was 6 - 13 mm (average: 10.2 mm) and huge in the control group. Also, as shown in Fig. 26, in pathological analysis, the inside of the seeded nodules in the GO-Y199-administered group had fallen into necrosis, while the control group was filled with tumor cells.

[0110] [Blood Concentration after Intravenous Injection] The blood concentration of GO-Y199 after intravenous injection was examined. The method was as follows. BALB / cSlc-nu / + mice were intravenously injected with the equivalent of 1 mg of GO-Y199 into the tail vein as described above, blood was collected from the orbital vein over time, and the serum was subjected to HPLC to measure the blood concentration of GO-Y199. The results are shown in Figs. 27 and 28. Fig. 27 is the result of HPLC analysis of blood after intravenous injection. Fig. 28 is the result of the change over time in the blood concentration of GO-Y199 after intravenous injection.

[0111] Fig. 27 is the raw data of HPLC analysis. As shown in Fig. 28, the blood concentration of GO-Y199 rapidly decreased until 60 minutes after intravenous injection, then decreased slowly, and GO-Y199 disappeared from the blood 180 minutes after intravenous injection. From this, it was confirmed that GO-Y199 can be administered intravenously, and the blood concentration is maintained for 3 hours after administration, and then it is rapidly metabolized and excreted.

[0112] [Safety and Toxicity Tests] The safety and toxicity of GO-Y199 in vivo were investigated. The methods were as follows. As described above, an equivalent of 1 mg of GO-Y199 was injected into the tail vein, and 7 days later, blood was collected from the orbital vein, and the hemoglobin value (Hb), white blood cell count (WBC), and platelet count (Plt) were measured using Cell tac MEK-5258 (manufactured by Nihon Kohden Corporation). The serum creatinine value (Cre), total bilirubin value (T-Bil), and AST value were measured externally (by SkyLight Biotech Co., Ltd.). The results are shown in Tables 12 and 13. Figure 29 shows the change in the body weight of the mice after intravenous injection. Figure 30 shows a photograph of the tails of the mice after intravenous injection.

[0113]

Table 12

[0114]

Table 13

[0115] As shown in Tables 12 and 13, no significant differences were confirmed between the control group and the GO-Y199 intravenous injection group. Also, as shown in Figure 29, no differences in body weight change were confirmed between the two groups. As shown in Figure 30, no local abnormalities were observed in the tails of all the mice after intravenous injection. From the above results, the safety of GO-Y199 for the mouse model was confirmed.

Claims

1. A compound represented by the following general formula or a salt thereof. 【Chemical Formula 1】 (R １ ~ R ４ is a hydrogen atom, C １－４ lower alkyl group, hydroxy C １－４ lower alkyl group, C １－４ lower alkoxy C １－４ lower alkyl group, and C １－４ lower alkoxy C １－４ lower alkoxy C １－４ lower alkyl group selected as a substituent, R １ ~ R ４ may be the same or different. n is 1 to 6. Sugar is a monosaccharide or a disaccharide.)

2. An antitumor agent comprising the compound according to claim 1 or a salt thereof as an active ingredient.

3. An antitumor agent for gastric cancer, colorectal cancer, pancreatic cancer, malignant mesothelioma, and cutaneous T-cell lymphoma, comprising the compound according to claim 1 or a salt thereof as an active ingredient.

4. An antitumor agent for pancreatic cancer, malignant mesothelioma, and cutaneous T-cell lymphoma, comprising the compound according to claim 1 or a salt thereof as an active ingredient.

5. An antitumor agent for pancreatic cancer, comprising the compound according to claim 1 or a salt thereof as an active ingredient.

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