A compound in salvia miltiorrhiza and a pharmaceutical composition and application of tumor immunotherapy thereof

Abstract

The application belongs to the field of medicine, and relates to a kind of salvia miltiorrhiza extract and pharmaceutical composition and application, specifically a kind of novel structure compound in salvia miltiorrhiza and preparation method and the application of the compound in tumor immunotherapy. Pharmacological test proves that the compound has significant inhibitory effect on AhR, and can be used for preparing a drug for tumor immunotherapy.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, and particularly relates to a kind of compounds isolated from traditional Chinese medicine Danshen and a pharmaceutical composition thereof in the application of tumor immunotherapy. BACKGROUND

[0002] Aryl hydrocarbon receptor (AhR) is a member of basic helix-loop-helix (bHLH) transcription factor family, and is a kind of ligand-dependent receptor. Different ligands have different affinities when combined with AhR. AhR ligands are diverse in types and sources, and can play an important role in cell cycle, body growth and development, immune cell differentiation, etc., especially in the regulation of tumor immunity.

[0003] Studies have shown that aryl hydrocarbon receptor can affect the growth, life, migration and invasion of tumors by participating in cell proliferation and apoptosis, immune metabolism, etc. When the expression of AhR is inhibited, the immune escape of tumor stem cells through the Kyn-AhR-PD-1 pathway can be blocked, thereby playing a killing role of T cells and drugs. Therefore, finding AhR inhibitors is a means to find anti-tumor drugs.

[0004] One of the most effective ways to find active compounds from traditional Chinese medicines as lead compounds to create new drugs is to find active compounds from traditional Chinese medicines as lead compounds to create new drugs. Danshen is the dried root and rhizome of Salvia miltiorrhiza Bunge of Lamiaceae (Lamiaceae) and is a perennial herb distributed in most areas of China. It has the traditional functions of activating blood and removing blood stasis, dredging channels and relieving pain, clearing heart and relieving restlessness, and cooling blood and resolving abscesses. Modern studies have shown that Danshen mainly contains liposoluble components such as tanshinone-type diterpenes and water-soluble components such as phenolic acids, and shows good activity in anti-tumor and neuroprotection. SUMMARY

[0005] Applicants have found a class of structurally novel compounds, Tanshinolic ester A-F, isolated from Danshen, which have the following chemical structures,

[0006]

[0007] The screening of AhR inhibitory activity of compound 3-4 showed that it had significant AhR inhibitory activity, with IC 50 of 1.25 μM and 1.51 μM. Currently, there is no research report on the use of such compounds as AhR inhibitors, preparation methods and patent documents.

[0008] To solve the technical problems of the present application, the present application provides the following technical solutions:

[0009] The first aspect of the technical scheme of the present application is to provide a kind of compound or its pharmaceutically acceptable salt, characterized in that the structure of the compound is as follows:

[0010]

[0011] The pharmaceutically acceptable salt described above is selected from the salt of compound and inorganic base, organic base. The organic base includes methylamine, ethylamine, diethylamine, triethylamine, propylamine, butylamine, octylamine, hexanediamine, ethylenediamine, propylenediamine, butylenediamine, benzylamine, phenethylamine, o-xylylamine, p-xylylamine, and the inorganic base includes sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide, zinc hydroxide, copper hydroxide, iron hydroxide, ammonium hydroxide, sodium bicarbonate and potassium bicarbonate.

[0012] Compound 1-6 are respectively extracted from Danshen, and Tanshinolic ester A is compound 1, Tanshinolic ester B is compound 2, Tanshinolic ester C is compound 3, Tanshinolic ester D is compound 4, Tanshinolic ester E is compound 5, and Tanshinolic ester F is compound 6; the basic skeleton structures of the six compounds are novel, and no literature and patent report the type of compound.

[0013] The second aspect of the technical scheme of the present application is to provide a preparation method of the compound in the first aspect, and the preparation method is as follows: 70 kg of Danshen medicinal materials are extracted by reflux extraction with 80% ethanol, the extract is concentrated, and then extracted with ethyl acetate; the solvent of the ethyl acetate part is recovered to obtain an extract, the extract is further subjected to silica gel column chromatography, eluted with a petroleum ether-acetone solvent system, and all eluted components are concentrated under reduced pressure to obtain 13 parts (Fr.1-Fr.13); Fra.9 is selected for further silica gel column chromatography, eluted with a petroleum ether-ethyl acetate solvent system to obtain 8 subparts (Fra 9.1-Fra 9.8); Fra 9.6 and Fra 9.7 are subjected to Sephadex LH-20 column chromatography to obtain fractions containing compounds Tanshinolic ester A-F, and the fractions are subjected to preparative high performance liquid purification; the structures of the fractions are analyzed and identified by UV, IR, NMR, MS and CD spectrum analysis, and the fractions are a type of quinone diterpene and styryl alcohol caffeic acid ester adduct; and currently, no such substance is reported.

[0014] The third aspect of the technical scheme of the present application provides a pharmaceutical composition, which comprises the compound of the first aspect of the present application or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient. The dosage form of the pharmaceutical composition includes tablets, capsules, pills, granules, oral liquids or suspensions.

[0015] The fourth aspect of the technical scheme of the present application provides the use of the compound of the first aspect of the present application in tumor immunotherapy.

[0016] Beneficial technical effects

[0017] 1. The novel compounds of Salvia miltiorrhiza in the present application have obvious AhR inhibitory activity and can be used for preparing drugs for tumor immunotherapy.

[0018] 2. The novel compounds of Salvia miltiorrhiza in the present application have novel structures and have not been reported in the literature. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 is Inhibitory activity of compounds 3 and 4 on AhR. DETAILED DESCRIPTION

[0020] The following examples and pharmacological activity experiments are used to further illustrate the present application, but this does not mean any limitation to the present application.

[0021] Example 1 Preparation and identification of monomer compounds Tanshinolic ester A-F in Salvia miltiorrhiza

[0022] 70 kg of Salvia miltiorrhiza Bge. was extracted with 80% ethanol for 3 times, 2 hours each time. The extract was concentrated and extracted with ethyl acetate for 3 times. The ethyl acetate fraction was recovered to obtain the extract, which was further purified by silica gel column chromatography, eluted with petroleum ether-acetone solvent system, with the elution ratio of 100:0, 80:1, 50:1, 40:1, 30:1, 20:1, 10:1, 5:1, 3:1, 2:1, 1:1, 0:100, and finally eluted with pure methanol. All the eluted fractions were concentrated under reduced pressure to obtain 13 fractions (Fr.1-Fr.13). Fra.9 was further purified by silica gel column chromatography, eluted with petroleum ether-ethyl acetate solvent system, with the elution ratio of 20:1, 15:1, 10:1, 5:1, 3:1, 2:1, 1:1, 0:1 to obtain 8 sub-fractions (Fra 9.1-Fra 9.8). Fra 9.6 was purified by Sephadex LH-20 column chromatography to obtain a fraction containing compounds Tanshinolic ester A-D, and Fra 9.7 was purified by Sephadex LH-20 column chromatography to obtain a fraction containing compounds Tanshinolic ester E-F, which were purified by preparative high performance liquid chromatography, respectively, with the following conditions: (1) column: YMC-Triart C18 (250 mm x 20 mm, 5 μm); (2) mobile phase: 0.2% acetic acid in water / methanol (60 / 40); (3) flow rate: 5.0 mL / min; (4) column temperature: 25 °C; (5) detection wavelength: 254 nm. The above compounds Tanshinolic ester A-F were obtained, which were analyzed and identified by UV, IR, NMR, MS and CD spectral methods to be a novel type of quinonoid diterpene and phenylethanoid caffeic acid ester adducts.

[0023] The spectral information and NMR signal assignment of the above novel compounds are as follows:

[0024] Tanshinolic ester A (Compound 1): white amorphous powder; (c 0.10, MeOH); UV (MeOH) λmax(log ε) 201 (3.99), 220 (3.68), 257 (3.93), 281 (3.51), 301 (3.51), 313 (3.51), 339 (3.36) nm; IR v max 3397, 2957, 2924, 1728, 1598, 1409, 1260, 1119, 1072, 1033, 802 cm -1 ; CD (MeOH) 226 (Δε -0.027), 250 (Δε 0.75), 285 (Δε 0.35), 329 (Δε -0.39) nm;1 H NMR (500 MHz, CD3OD) δ H : 9.57 (1H, dd, J = 3.0 Hz, 7.0 Hz, H-1), 7.87 (1H, d, J = 9.0 Hz, H-6), 7.71 (1H, d, J = 9.0 Hz, H-7), 7.62 (1H, d, J = 16.0 Hz, H-25), 7.42 (1H, s, H-14), 7.40 (1H, overlap, H-3), 7.39 (1H, overlap, H-2), 7.01 (1H, J = 2.0 Hz, H-20), 7.00 (1H, J = 2.0 Hz, H-31), 6.92 (dd, J = 2.0 Hz, 8.0 Hz, H-24), 6.89 (dd, J = 2.0 Hz, 8.0 Hz, H-35), 6.77 (1H, d, J = 8.0 Hz, H-34), 6.74 (1H, d, J = 8.0 Hz, H-23), 6.71 (1H, d, J = 4.0 Hz, H-28), 6.28 (1H, d, J = 16.0 Hz, H-26), 5.39 (1H, d, J = 4.0 Hz, H-29), 3.37 (1H, m, H-15), 2.73 (3H, s, H-18), 1.32 (3H, d, J = 7.0 Hz, H-16), 1.31 (3H, d, J = 7.0 Hz, H-17); 13 C NMR (125 MHz, CD3OD) δ C : 167.2 (C-27), 150.1 (C-22), 148.9 (C-25), 147.0 (C-33), 146.8 (C-21), 146.6 (C-32), 141.7 (C-11), 138.3 (C-12), 138.3 (C-13), 134.9 (C-4), 132.6 (C-5), 131.1 (C-10), 129.9 (C-8), 128.8 (C-30), 128.2 (C-3), 128.0 (C-7), 127.7 (C-1), 127.4 (C-19), 126.5 (C-2), 123.5 (C-24), 122.6 (C-6), 120.6 (C-9), 119.5 (C-14), 116.5 (C-34), 116.4 (C-23), 115.3 (C-20), 114.9 (C-31), 113.7 (C-26), 91.1 (C-28), 76.6 (C-29), 28.6 (C-15), 23.0 (C-17), 22.7 (C-16), 20.6 (C-18); HR-ESI-MS m / z 578.1941 [M+H] + (calcd for C35 H 30 O8,578.6170).

[0025] Tanshinolic ester B (Compound 2): white amorphous powder; (c 0.10, MeOH); UV (MeOH) λmax(log ε) 201 (3.99), 220 (3.68), 257 (3.93), 281 (3.51), 301 (3.51), 313 (3.51), 339 (3.36) nm; IR v max 3397, 2957, 2924, 1728, 1598, 1409, 1260, 1119, 1072, 1033, 802 cm -1 ; CD (MeOH) 221 (Δε 0.26), 249 (Δε -0.48), 281 (Δε -0.083), 323 (Δε 0.39) nm; 1 H NMR (500 MHz, CD3OD) δ H : 9.57 (1H, dd, J = 3.0 Hz, 7.0 Hz, H-1), 7.87 (1H, d, J = 9.0 Hz, H-6), 7.71 (1H, d, J = 9.0 Hz, H-7), 7.62 (1H, d, J = 16.0 Hz, H-25), 7.42 (1H, s, H-14), 7.40 (1H, overlap, H-3), 7.39 (1H, overlap, H-2), 7.01 (1H, J = 2.0 Hz, H-20), 7.00 (1H, J = 2.0 Hz, H-31), 6.92 (dd, J = 2.0 Hz, 8.0 Hz, H-24), 6.89 (dd, J = 2.0 Hz, 8.0 Hz, H-35), 6.77 (1H, d, J = 8.0 Hz, H-34), 6.74 (1H, d, J = 8.0 Hz, H-23), 6.71 (1H, d, J = 4.0 Hz, H-28), 6.28 (1H, d, J = 16.0 Hz, H-26), 5.39 (1H, d, J = 4.0 Hz, H-29), 3.37 (1H, m, H-15), 2.73 (3H, s, H-18), 1.32 (3H, d, J = 7.0 Hz, H-16), 1.31 (3H, d, J = 7.0 Hz, H-17); 13 C NMR (125 MHz, CD3OD) δ C:167.2 (C-27), 150.1 (C-22), 148.9 (C-25), 147.0 (C-33), 146.8 (C-21), 146.6 (C-32), 141.7 (C-11), 138.3 (C-12), 138.3 (C-13), 134.9 (C-4), 132.6 (C-5), 131.1 (C-10), 129.9 (C-8), 128.8 (C-30), 128.2 (C-3), 128.0 (C-7), 127.7 (C-1), 127.4 (C-19), 126.5 (C-2), 123.5 (C-24), 122.6 (C-6), 120.6 (C-9), 119.5 (C-14), 116.5 (C-34), 116.4 (C-23), 115.3 (C-20), 114.9 (C-31), 113.7 (C-26), 91.1 (C-28), 76.6 (C-29), 28.6 (C-15), 23.0 (C-17), 22.7 (C-16), 20.6 (C-18); HR-ESI-MS m / z 578.1941 [M+H] + (calcd for C 35 H 30 O8,578.6170).

[0026] Tanshinolic ester C (Compound 3): white amorphous powder; (c 0.10, MeOH); UV (MeOH) λmax(log ε) 201 (4.01), 220 (3.68), 257 (3.90), 281 (3.49), 301 (3.45), 313 (3.49), 339 (3.33) nm; IR v max 3412, 2958, 2852, 1728, 1593, 1410, 1260, 1098, 1032, 801 cm -1 ; CD (MeOH) 217 (Δε 5.34), 233 (Δε -10.15), 255 (Δε 51.24), 299 (Δε 13.93) nm, 332 (Δε -8.53) nm; 1 H NMR (500 MHz, CD3OD) δ H9.37 (1H, dd, J = 2.0 Hz, 8.0 Hz, H-1), 7.84 (1H, d, J = 9.0 Hz, H-6), 7.70 (1H, d, J = 9.0 Hz, H-7), 7.63 (1H, d, J = 16.0 Hz, H-25), 7.47 (1H, s, H-14), 7.39 (1H, overlap, H-2), 7.38 (1H, overlap, H-3), 7.02 (1H, J = 2.0 Hz, H-20), 6.97 (1H, J = 2.0 Hz, H-31), 6.92 (dd, J = 2.0 Hz, 8.0 Hz, H-24), 6.88 (dd, J = 2.0 Hz, 8.0 Hz, H-35), 6.78 (1H, d, J = 8.0 Hz, H-34), 6.74 (1H, d, J = 8.0 Hz, H-23), 6.69 (1H, d, J = 4.0 Hz, H-28), 6.28 (1H, d, J = 16.0 Hz, H-26), 5.23 (1H, d, J = 4.0 Hz, H-29), 3.50 (1H, m, H-15), 2.70 (3H, s, H-18), 1.39 (3H, d, J = 7.0 Hz, H-16), 1.36 (3H, d, J = 7.0 Hz, H-17); 13 C NMR (125 MHz, CD3OD) δ C : 167.2 (C-27), 150.1 (C-22), 149.0 (C-25), 147.1 (C-33), 146.8 (C-21), 146.6 (C-32), 140.4 (C-12), 139.5 (C-11), 138.2 (C-13), 134.8 (C-4), 132.6 (C-5), 130.7 (C-10), 129.5 (C-8), 128.9 (C-30), 128.2 (C-3), 127.9 (C-7), 127.6 (C-1), 127.4 (C-19), 126.5 (C-2), 123.5 (C-24), 122.4 (C-6), 120.7 (C-9), 120.0 (C-14), 116.5 (C-23), 116.4 (C-34), 115.3 (C-20), 115.1 (C-31), 113.7 (C-26), 91.4 (C-28), 76.6 (C-29), 28.6 (C-15), 23.0 (C-17), 22.7 (C-16), 20.6 (C-18); HR-ESI-MS m / z 578.1941 [M+H] + (calcd for C 35 H 30 O8, 578.6170).

[0027] Tanshinolic ester D (Compound 4): white amorphous powder; (c 0.10, MeOH); UV (MeOH) λmax(log ε) 201 (4.01), 220 (3.68), 257 (3.90), 281 (3.49), 301 (3.45), 313 (3.49), 339 (3.33) nm; IR v max 3412, 2958, 2852, 1728, 1593, 1410, 1260, 1098, 1032, 801 cm -1 ; CD (MeOH) 217 (Δε -2.35), 233 (Δε 10.82), 255 (Δε -46.33), 299 (Δε -11.72) nm, 332 (Δε 9.08) nm; 1 H NMR (500 MHz, CD3OD) δ H : 9.37 (1H, dd, J = 2.0 Hz, 8.0 Hz, H-1), 7.84 (1H, d, J = 9.0 Hz, H-6), 7.70 (1H, d, J = 9.0 Hz, H-7), 7.63 (1H, d, J = 16.0 Hz, H-25), 7.47 (1H, s, H-14), 7.39 (1H, overlap, H-2), 7.38 (1H, overlap, H-3), 7.02 (1H, J = 2.0 Hz, H-20), 6.97 (1H, J = 2.0 Hz, H-31), 6.92 (dd, J = 2.0 Hz, 8.2 Hz, H-24), 6.88 (dd, J = 2.0 Hz, 8.0 Hz, H-35), 6.78 (1H, d, J = 8.0 Hz, H-34), 6.74 (1H, d, J = 8.0 Hz, H-23), 6.69 (1H, d, J = 4.0 Hz, H-28), 6.28 (1H, d, J = 16.0 Hz, H-26), 5.23 (1H, d, J = 4.0 Hz, H-29), 3.50 (1H, m, H-15), 2.70 (3H, s, H-18), 1.39 (3H, d, J = 7.0 Hz, H-16), 1.36 (3H, d, J = 7.0 Hz, H-17); 13 C NMR (125 MHz, CD3OD) δ C:167.2 (C-27), 150.1 (C-22), 149.0 (C-25), 147.1 (C-33), 146.8 (C-21), 146.6 (C-32), 140.4 (C-12), 139.5 (C-11), 138.2 (C-13), 134.8 (C-4), 132.6 (C-5), 130.7 (C-10), 129.5 (C-8), 128.9 (C-30), 128.2 (C-3), 127.9 (C-7), 127.6 (C-1), 127.4 (C-19), 126.5 (C-2), 123.5 (C-24), 122.4 (C-6), 120.7 (C-9), 120.0 (C-14), 116.5 (C-23), 116.4 (C-34), 115.3 (C-20), 115.1 (C-31), 113.7 (C-26), 91.4 (C-28), 76.6 (C-29), 28.6 (C-15), 23.0 (C-17), 22.7 (C-16), 20.6 (C-18); HR-ESI-MS m / z 578.1941 [M+H] + (calcd for C 35 H 30 O8,578.6170).

[0028] Tanshinolic ester E (Compound 5): white amorphous powder; (c 0.10, MeOH); UV (MeOH) λmax(log ε) 201 (3.94), 220 (3.69), 257 (3.91), 281 (3.53), 301 (3.49), 313 (3.51), 339 (3.37) nm; IR v max 3403, 2958, 2826, 1722, 1627, 1598, 1409, 1263, 1118, 1018, 802 cm -1 ; CD (MeOH) 217 (Δε -2.45), 235 (Δε -3.20), 257 (Δε 9.85), 304 (Δε 3.49) nm, 337 (Δε -0.39) nm; 1 H NMR (500 MHz, CD3OD) δ H9.37 (1H, dd, J = 2.5 Hz, 7.5 Hz, H-1), 7.86 (1H, d, J = 9.0 Hz, H-6), 7.71 (1H, d, J = 9.0 Hz, H-7), 7.63 (1H, d, J = 16.0 Hz, H-25), 7.50 (1H, s, H-14), 7.39 (1H, overlap, H-2), 7.39 (1H, overlap, H-3), 7.02 (1H, J = 2.0 Hz, H-20), 6.97 (1H, J = 2.0 Hz, H-31), 6.94 (dd, J = 2.0 Hz, 8.0 Hz, H-24), 6.89 (dd, J = 2.1 Hz, 8.0 Hz, H-35), 6.79 (1H, d, J = 8.0 Hz, H-34), 6.75 (1H, d, J = 8.0 Hz, H-23), 6.71 (1H, d, J = 4.0 Hz, H-28), 6.27 (1H, d, J = 16.0 Hz, H-26), 5.25 (1H, d, J = 4.2 Hz, H-29), 3.95 (1H, d, J = 11.0 Hz, J = 7.5 Hz, H-16a), 3.71 (3H, d, J = 11.0 Hz, J = 6.0 Hz, H-16b), 3.57 (1H, m, H-15), 2.71 (3H, s, H-18), 1.41 (3H, d, J = 7.0 Hz, H-17); 13 C NMR (125 MHz, CD3OD) δ C : 167.1 (C-27), 150.1 (C-22), 149.0 (C-25), 147.2 (C-33), 146.8 (C-21), 146.7 (C-32), 140.7 (C-12), 139.7 (C-11), 134.9 (C-4), 133.9 (C-13), 132.7 (C-5), 130.7 (C-10), 129.5 (C-8), 128.8 (C-30), 128.4 (C-3), 127.8 (C-7), 127.6 (C-1), 127.4 (C-19), 126.6 (C-2), 123.5 (C-24), 122.5 (C-6), 121.6 (C-14), 121.1 (C-9), 116.5 (C-34), 116.4 (C-23), 115.3 (C-31), 115.1 (C-20), 113.7 (C-26), 91.5 (C-28), 76.7 (C-29), 67.6 (C-16), 37.0 (C-15), 20.5 (C-18); 17.3 (C-17), HR-ESI-MS m / z 594.1890 [M+H] + (calcd for C35 H 30 O9,594.6160).

[0029] Tanshinolic ester F (Compound 6): white amorphous powder; (c 0.10, MeOH); UV (MeOH) λmax(log ε) 201 (3.94), 220 (3.69), 257 (3.92), 281 (3.53), 301 (3.49), 313 (3.52), 339 (3.37) nm; IR v max 3403, 2959, 2927, 2858, 1727, 1656, 1599, 1409, 1262, 1119, 1070, 802 cm -1 ; CD (MeOH) 217 (Δε -2.45), 235 (Δε -3.20), 257 (Δε 9.85), 304 (Δε 3.49) nm, 337 (Δε -0.39) nm; 1 H NMR (500 MHz, CD3OD) δ H : 9.37 (1H, dd, J = 3.0 Hz, 7.0 Hz, H-1), 7.86 (1H, d, J = 9.0 Hz, H-6), 7.71 (1H, d, J = 9.0 Hz, H-7), 7.63 (1H, d, J = 16.0 Hz, H-25), 7.50 (1H, s, H-14), 7.39 (1H, overlap, H-2), 7.39 (1H, overlap, H-3), 7.02 (1H, J = 2.0 Hz, H-20), 6.97 (1H, J = 2.0 Hz, H-31), 6.94 (dd, J = 2.0 Hz, 8.0 Hz, H-24), 6.89 (dd, J = 2.0 Hz, 8.0 Hz, H-35), 6.79 (1H, d, J = 8.0 Hz, H-34), 6.754 (1H, d, J = 8.0 Hz, H-23), 6.71 (1H, d, J = 4.0 Hz, H-28), 6.27 (1H, d, J = 16.0 Hz, H-26), 5.25 (1H, d, J = 4.0 Hz, H-29), 3.95 (1H, d, J = 11.0 Hz, J = 7.5 Hz, H-16a), 3.71 (3H, d, J = 11.0 Hz, J = 6.0 Hz, H-16b), 3.58 (1H, m, H-15), 2.71 (3H, s, H-18), 1.41 (3H, d, J = 7.0 Hz, H-17); 13 C NMR (125 MHz, CD3OD) δ C:167.2(C-27), 150.1(C-22), 149.0(C-25), 147.2(C-33), 146.8(C-21), 146.6(C-32), 140.7(C-12), 139.6(C-11), 134.9(C-4), 133.8(C-13), 132.7(C-5), 130.7(C-10), 129.5(C-8), 128.8(C-30), 128.4(C-3), 127.8(C-7), 127.6(C-1), 127.5(C-19), 126.6(C-2), 123.5(C-24), 122.5(C-6), 121.7(C-14), 121.1(C-9), 116.5(C-34), 116.4(C-23), 115.4(C-31), 115.2(C-20), 113.7(C-26), 91.4(C-28), 76.7(C-29), 67.6(C-16), 37.0(C-15), 20.5(C-18); 17.3(C-17), HR-ESI-MS m / z 594.1890 [M+H] + (calcd for C 35 H 30 O9, 594.6160).

[0030] AhR inhibitory activity assay of compounds 3 and 4 in Example 2

[0031] The dioxin-responsive element (DRE) and CYP1A1 core promoter sequence were constructed into the reporter gene plasmid (pGL3-basic) to obtain the recombinant vector pCL-CR2. Hepa 1 cells were transfected with pCL-CR2, and the stably transfected cell line CBG2.8D was obtained by G418 resistance screening.

[0032] CBG2.8D stable cell line was inoculated in 96-well white plates at a concentration of 4 x 10 5 After 24 hours of culture, the supernatant was aspirated and replaced with medium containing different concentrations of samples, with 3 wells in each group as controls. After 24 hours of continuous culture, the culture solution was discarded. Each well was washed once with 100 μL of PBS, and after the PBS was removed, 50 ul of luciferase lysis solution was added, shaken at room temperature for 20 min, and then detected for luciferase intensity.

[0033] Compounds 3 and 4 were screened for AhR inhibitory activity, and the results of the half-effective concentration are shown in Table 1. Figure 1.

[0034]

Claims

1. A class of compounds or pharmaceutically acceptable salts thereof, characterized in that, The structure of the compound is as follows:

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, The pharmaceutically acceptable salt is selected from the salt of the compound and inorganic base or organic base.

3. The compound according to claim 2, or a pharmaceutically acceptable salt thereof, characterized in that, The organic base is methylamine, ethylamine, diethylamine, triethylamine, propylamine, butylamine, octylamine, hexanediamine, ethylenediamine, propylenediamine, butylenediamine, benzylamine, phenethylamine, o-xylylamine, p-xylylamine, and the inorganic base includes sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide, zinc hydroxide, copper hydroxide, iron hydroxide, ammonium hydroxide, sodium bicarbonate and potassium bicarbonate.

4. Process for the preparation of a compound according to claim 1, characterized in that, The preparation method is as follows: 80% ethanol reflux extraction of Danshen medicinal materials, concentration, and then extract by organic solvent extraction, silica gel column chromatography, reverse phase silica gel column chromatography and preparative HPLC separation and purification to obtain the compound 1-4 in claim 1.

5. A pharmaceutical composition, characterized by, The pharmaceutical composition comprises the compound or its pharmaceutically acceptable salt in any one of claims 1-3 and pharmaceutically acceptable carrier or excipient.

6. Pharmaceutical composition according to claim 5, characterized in that The dosage form of the pharmaceutical composition includes tablets, capsules, pills, granules, oral liquid or suspension.

7. Use of the compound or its pharmaceutically acceptable salt in any one of claims 1-3 in the preparation of tumor immunological drugs.

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