Licoagroside B derivative, and preparation method, pharmaceutical composition and application thereof

Abstract

The invention belongs to the technical field of medicines, particularly discloses a class of licoagroside B derivatives and a preparation method, a pharmaceutical composition and application thereof, and particularly relates to a compound as shown in a formula (I) or pharmaceutically acceptable salt thereof and a preparation method thereof, and a composition containing the compound, the compound has the effect of remarkably reversing polarization activity of M2 macrophages, and has the effect of remarkably reversing polarization activity of M2 macrophages. Therefore, the compound can be used for preparing drugs for resisting tumors, fibrosis diseases, viruses, bacteria, infection and other related diseases. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of medicine, and in particular relates to a class of licoagroside B derivatives and application thereof in reversing M2 macrophage polarization. Background Art

[0002] When performing specific functions, macrophages will polarize under different physiological conditions and tissue microenvironments. According to the different biological functions they perform, they can be roughly divided into M1 and M2 types. They play an important role in many diseases, including tumors, infections (bacteria and viruses) and various fibrotic diseases. The main function of M1 is to activate immune response to respond to the invasion of foreign antigens and to eliminate abnormal cells in the body. M2 mainly mediates immune tolerance and damage repair.

[0003] During tumor occurrence and progression, tumor-associated macrophages (TAMs) are abundantly infiltrated in the tumor microenvironment, which has an important impact on tumor metastasis, angiogenesis, and immune escape. At the same time, the high density of TAMs in tumors of most patients is closely related to their poor prognosis. Therefore, TAMs targeted therapy is a hot strategy in anti-tumor treatment. In the tumor microenvironment, macrophages are mainly M2 type according to their phenotype. The M2 phenotype can promote tumor cell growth, while the M1 phenotype can inhibit tumor growth and stimulate anti-tumor immune response. Therefore, it is an important direction for the development of tumor immunotherapy to find drugs that inhibit M2 macrophages in the tumor microenvironment, induce them to repolarize from M2 to M1, and then enhance the inhibitory effect of macrophages on tumor cells. In addition, in non-tumor diseases, the infiltration of M2 macrophages in diseased organs is also an important cause of fibrosis caused by many organ damage, such as end-stage fibrosis of the lungs, liver, or kidneys. Therefore, reversing the increase of M2 macrophages is also one of the important means to improve fibrotic diseases. Furthermore, reversing M2 and increasing M1 macrophages is also an important means to enhance immunity and fight against infectious diseases such as bacteria, viruses, and mycoplasma.

[0004] Sophora tonkinensis is taken from the dried roots and rhizomes of the leguminous plant Sophora tonkinensis Gapnea. The Kaibao Materia Medica records that Sophora tonkinensis has the effects of "detoxifying various drugs, relieving pain, eliminating sores and swelling, treating acute jaundice, fever and cough, and killing small insects." The Bencao Qiuzhen calls Sophora tonkinensis "the first medicine for purging the heart and protecting the lungs, and reducing the reverse fire of the yin meridians, and relieving sore throat." Therefore, it is generally believed that Sophora tonkinensis has the effects of clearing heat and detoxifying, reducing swelling and relieving sore throat. It is a traditional Chinese medicine used to treat sore throat, swollen gums, hepatitis, cancer and other diseases. According to literature reports, more than 200 natural products have been found from Sophora tonkinensis, the main components of which are alkaloids and flavonoid compounds, which have anti-tumor, anti-inflammatory, liver protection and antiviral biological activities.

[0005] This patent isolated a novel class of licoagroside B derivatives from Sophora flavescens. Pharmacological experiments have demonstrated that they can inhibit the polarization of M2 macrophages and promote their polarization towards M1 macrophages, that is, they can induce macrophages in the tumor microenvironment to repolarize from M2 to M1, thereby enhancing the phagocytic ability of macrophages against tumor cells. Currently, there are no research reports on the reversal of M2 macrophage polarization by licoagroside B derivatives. Summary of the Invention

[0006] The technical problem solved by the present invention is to provide a new class of licoagroside B derivatives, their preparation methods, and pharmaceutical compositions, which can play roles in anti-tumor cell growth, anti-fibrotic diseases, anti-virus, anti-bacteria, anti-infection, etc. by reversing the polarization of M2 macrophages and promoting macrophage phagocytosis, and their applications in the preparation of drugs for treating the above diseases.

[0007] A series of licoagroside B derivatives were found in Sophora flavescens. These compounds all have the structure of licoagroside B, that is, maltol glycoside is esterified with 3-methyl-3-hydroxyglutaric acid. The other carboxylic acid of 3-methyl-3-hydroxyglutaric acid is esterified with glucose in flavonoid glycosides, lignan glycosides, etc. to form a double glycoside ester structure.

[0008] The first aspect of the technical solution of the present invention is to provide a compound of formula (Ⅰ) or a pharmaceutically acceptable salt thereof,

[0009]

[0010] wherein R1 is independently selected from flavonoids and their glycosides, isoflavonoids and their glycosides, phenylpropanoids and their glycosides, lignans and their glycosides, ionol alcohols and their glycosides; R2 is independently selected from H, apiose; is independently selected from

[0011] Preferably, wherein R1 has a fragment of formula II,

[0012]

[0013] The parent nucleus I can be independently linked to the A1, A2, or A3 position of formula II; wherein A1, A2, and A3 are independently selected from H, OH, C1-C3 alkoxy groups, or monosaccharides, and the alkoxy group is preferably a straight-chain or branched-chain alkoxy group, further preferably OCH3 or OC2H5, and the monosaccharide is preferably glucose, xylose, or apiose.

[0014] Alternatively, preferably, R1 has a fragment of formula III,

[0015]

[0016] The parent nucleus I can be independently linked to the B1, B2, B3 or B4 position of Formula III; B1, B2, B3 and B4 are independently selected from H, OH, alkoxy groups having 1 to 3 carbon atoms or monosaccharides, wherein the alkoxy group is preferably a straight-chain or branched-chain alkoxy group, more preferably OCH3 or OC2H5, and the monosaccharide is preferably glucose, xylose or apiose.

[0017] Alternatively, preferably, R1 has a fragment of Formula IV,

[0018]

[0019] The parent nucleus I can be independently linked to the C1 or C2 position of Formula IV; C1 and C2 are independently selected from H, OH, alkoxy groups having 1 to 3 carbon atoms or monosaccharides, wherein the alkoxy group is preferably a straight-chain or branched-chain alkoxy group, more preferably OCH3 or OC2H5, and the monosaccharide is preferably glucose, xylose or apiose; independently selected from

[0020] Alternatively, preferably, R1 has a fragment of Formula V,

[0021]

[0022] The parent nucleus I can be independently linked to the D1, D2, D3 or D4 position of Formula V; D1, D2, D3 and D4 are independently selected from H, OH, alkoxy groups having 1 to 3 carbon atoms or monosaccharides, wherein the alkoxy group is preferably a straight-chain or branched-chain alkoxy group, more preferably OCH3 or OC2H5, and the monosaccharide is preferably glucose, xylose or apiose; independently selected from

[0023] Alternatively, preferably, R1 has a fragment of Formula VI,

[0024]

[0025] The parent nucleus I can be independently linked to the E1, E2, E3 or E4 position of Formula VI; E1, E2, E3 and E4 are independently selected from H, OH, alkoxy groups having 1 to 3 carbon atoms or monosaccharides, wherein the alkoxy group is preferably a straight-chain or branched-chain alkoxy group, more preferably OCH3 or OC2H5, and the monosaccharide is preferably glucose, xylose or apiose; independently selected from

[0026] Alternatively, preferably, R1 has a fragment of Formula VII,

[0027]

[0028] The mother nucleus I can be independently connected to the F1, F2, F3 or F4 positions of Formula VII; F1, F2, F3 and F4 are independently selected from H, OH, alkoxy groups with 1 to 3 carbon atoms or monosaccharides, where the alkoxy groups are preferably straight-chain or branched-chain alkoxy groups, more preferably OCH3 or OC2H5, and the monosaccharides are preferably glucose, xylose or apiose; independently selected from

[0029] Alternatively, preferably, R1 has a fragment of Formula VIII,

[0030]

[0031] The mother nucleus I can be independently connected to the G1, G2, G3 or G4 positions of Formula VIII; G1, G2, G3 and G4 are independently selected from H, OH, alkoxy groups with 1 to 3 carbon atoms or monosaccharides, where the alkoxy groups are preferably straight-chain or branched-chain alkoxy groups, more preferably OCH3 or OC2H5, and the monosaccharides are preferably glucose, xylose or apiose; independently selected from

[0032] Alternatively, preferably, R1 has a fragment of Formula IX,

[0033]

[0034] The mother nucleus I can be independently connected to the K1, K2 or K3 positions of Formula IX; K1, K2 and K3 are independently selected from H, OH, alkoxy groups with 1 to 3 carbon atoms, where the alkoxy groups are preferably straight-chain or branched-chain alkoxy groups, more preferably OCH3 or OC2H5, and the monosaccharides are preferably glucose, xylose or apiose.

[0035] Furthermore, the compounds are selected from:

[0036]

[0037] The pharmaceutically acceptable salts of the above compounds are selected from the salts formed by the compounds and organic bases, inorganic bases. The organic bases include meglumine, tris(hydroxymethyl)aminomethane, N-methyl-glucosamine, methylamine, ethylamine, diethylamine, triethylamine, propylamine, butylamine, octylamine, hexamethylenediamine, ethylenediamine, propylenediamine, butylenediamine, benzylamine, phenethylamine, o-phthalylenediamine, p-phthalylenediamine, and the inorganic bases include sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide, zinc hydroxide, copper hydroxide, iron hydroxide, ammonium hydroxide, sodium bicarbonate, potassium bicarbonate, sodium carbonate, potassium carbonate.

[0038] The second aspect of the technical solution of the present invention is to provide a preparation method of the compound described in the first aspect or a pharmaceutically acceptable salt thereof, and the preparation method is as follows: The Sophora tonkinensis root medicinal material is crushed and extracted with a solvent. After the extract is concentrated, the above-mentioned compound is obtained through organic solvent extraction and column chromatography separation.

[0039] The extraction solvent described above includes water, ethanol, and ethanol aqueous solutions with any concentration.

[0040] The extraction solvent is an organic solvent immiscible with water or a mixed solvent in any proportion. The organic solvents include, but are not limited to, n-butanol, ethyl acetate, chloroform, dichloromethane, ether, petroleum ether, cyclohexane, n-hexane. The mixed solvents include, but are not limited to, mixed liquids of two or more of the above-mentioned organic solvents in any proportion and mixed liquids of two or more of the above-mentioned solvents and solvents such as methanol, ethanol, and acetone in any proportion.

[0041] The column chromatography includes normal pressure column chromatography, medium pressure column chromatography, and high performance liquid column chromatography. The fillers used include normal phase fillers, reverse phase fillers, gel fillers, and ion exchange resins. The normal phase fillers include fillers such as silica gel, diol, cyano, and amino; the reverse phase fillers include macroporous resins, microporous resins, C18, C8, phenyl, etc.; the gel fillers include HW type gels and Sephadex type gels.

[0042] The third aspect of the technical solution of the present invention provides a pharmaceutical composition, and the pharmaceutical composition includes one or more compounds in an effective dose described in the first aspect of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient. The dosage forms of the pharmaceutical composition include tablets, capsules, pills, granules, powders, dripping pills, oral liquids, or suspensions. The pharmaceutical composition can be made into conventional preparations, or can also be sustained release preparations, controlled release preparations, targeted preparations, and various particulate drug delivery systems.

[0043] In order to formulate the compounds of the present invention into tablets, various excipients well-known in the art can be widely used, including diluents, binders, wetting agents, disintegrants, lubricants, and glidants. The diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the wetting agents can be water, ethanol, isopropanol, etc.; the binders can be starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, acacia mucilage, gelatin mucilage, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; the disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitan fatty acid ester, sodium dodecyl sulfate, etc.; the lubricants and glidants can be talc, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc. The tablets can be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer tablets and multilayer tablets. In order to formulate the dosage unit into capsules, the active ingredient, the compound of the present invention, can be mixed with a diluent and a glidant, and the mixture can be directly placed into hard capsules or soft capsules. The various diluents, binders, wetting agents, disintegrants, and glidants used for preparing the tablets of the compounds of the present invention can also be used for preparing the capsules of the compounds of the present invention. In addition, if necessary, colorants, preservatives, fragrances, flavoring agents, or other additives can also be added to the pharmaceutical preparation. For the purpose of achieving the therapeutic effect and enhancing the treatment effect, the drugs or pharmaceutical compositions of the present invention can be administered by any well-known administration methods.

[0044] The fourth aspect of the technical solution of the present invention is to provide the use of the compound described in the first aspect of the present invention or its pharmaceutically acceptable salt in the preparation of drugs for treating anti-tumor, anti-fibrotic diseases, anti-viral, anti-bacterial, anti-infectious and other diseases. The described diseases that can be treated include tumors, organ fibrosis, various infections (including viruses, bacteria, and mycoplasmas), etc.

[0045] Beneficial technical effects:

[0046] A novel class of licoagroside B derivatives discovered from Sophora flavescens Ait. in the present invention has a significant effect of inhibiting the polarization of M2 macrophages, can promote the phagocytosis of tumor cells by macrophages, and thus play an anti-tumor therapeutic role. The role of reversing M2 macrophage polarization is also related to the effects of anti-fibrosis of various organs and anti-infection. Therefore, this class of components has the application prospect of being further developed into anti-tumor, anti-fibrotic disease, anti-viral, anti-bacterial, and anti-infectious drugs. Detailed implementation manners

[0047] The following examples and pharmacological activity experiments are used to further illustrate the present invention, but this does not mean any limitation to the present invention.

[0048] Example Preparation of licoagroside B derivatives from Sophora tonkinensis

[0049] The Sophora tonkinensis medicinal materials (50 kg) were crushed and refluxed with distilled water for extraction three times, 2 hours each time. The extraction solution was concentrated and subjected to macroporous resin column chromatography, eluted successively with ethanol (0 - 95%). The fraction eluted with 30% ethanol was concentrated and then passed through an MCI column, eluted successively with ethanol (0 - 75%). The fractions eluted with 10% and 20% ethanol were passed through a silica gel (60 - 100 mesh) column and eluted with a chloroform - methanol (99:1 - 1:1) gradient to obtain Fr.1 - 41 and Fr.G - 1 - 32. Fr.18 - 24 and Fr.G - 11 - 18 were combined and eluted through a Diol Flash column (CHCl3 - MeOH, 20:1 - 2:1) to obtain M7 - D1 - 26.

[0050] 1) Fraction M7 - D4 - 6 was eluted through a Diol Flash column (EtOAc - MeOH, 30:1 - 5:1) to obtain Fr.D - 1 - 22.

[0051] Fraction Fr.D - 5 was passed through a Toyopearl HW - 40F column (MeOH - H2O, 90%) to obtain 3 components Fr.D - 5 - 1 to 3. Fr.D - 5 - 1 was passed through a semi - preparative PFP chromatographic column (2.5 mL / min, 50% methanol, wavelength 205 nm) to obtain compound 7 (t R = 20.5 min) and 8 (t R = 26.0 min); Fr.D - 5 - 2 was passed through a semi - preparative PFP chromatographic column (2.5 mL / min, wavelength 205 nm) to obtain compound 6 (60% methanol, t R = 12.5 min), 9 (49% methanol, t R = 27.0 min). Fraction Fr.D - 6 was eluted through a C18 Flash column (MeOH - H2O, 20 - 60%) to obtain 3 components. Component 1 was passed through a semi - preparative MG - Ⅱ chromatographic column (2.5 mL / min, 50% methanol, wavelength 205 nm) to obtain compound 4 (t R = 20.5 min); Component 2 was passed through a semi - preparative PFP chromatographic column (2.5 mL / min, wavelength 205 nm) to obtain compound 1 (60% methanol, t R = 17.5 min), 2 (58% methanol, t R = 25.0 min), 5 (60% methanol, t R= 14.0 min); Component 3 used a semi-preparative C8 chromatographic column (2.5 mL / min, wavelength 205 nm) to obtain Compound 10 (40% methanol, t R = 45.0 min) and 11 (40% methanol, t R = 46.3 min).

[0052] Fraction Fr.D-7 used a semi-preparative PFP chromatographic column (2.5 mL / min, 54% methanol, wavelength 205 nm) to obtain Compound 3 (t R = 31.0 min).

[0053] Fraction Fr.D-8 used a semi-preparative PFP chromatographic column (2.5 mL / min, 40% methanol, wavelength 205 nm) to obtain 13 (t R = 50.0 min).

[0054] 2) From fraction M7-D7, using a PFP semi-preparative chromatographic column (2.5 mL / min, 49% methanol, wavelength 205 nm), Compound 12 (t R = 21.2 min) was obtained.

[0055] 3) From fraction M7-D9, using a semi-preparative C8 chromatographic column (35% methanol, 2.5 ml / min, wavelength 205 nm), Compound 14 (t R = 49.8 min), 15 (t R = 49.2 min) were obtained.

[0056] The structures of the compounds were determined by spectroscopic methods such as UV, IR, NMR, MS and CD.

[0057] The spectral information and NMR signal assignments of the above new compounds are as follows:

[0058] Compound 1 Maltol [5”-O-6””'-(4””,7”'-dihydroxy-3””-methoxyflavone-4””-O-β-D-glucopyranosyl)-6'-O-(3”-hydroxy-3”-methylglutaryl)]-3-O-β-D-glucopyranoside

[0059] Pale yellow powder; ECD (MeOH) λ max (Δε) 215 (-3.41), 318 (-0.80) nm; UV (MeOH) λ max (logε) 208 (4.47), 235 (4.17), 328 (4.06) nm; IR ν max3366, 2923, 2852, 1731, 1627, 1601, 1508, 1454, 1381, 1254, 1073 cm -1 ; 1 H NMR and 13 C NMR are shown in Tables 1-1 and 1-2; (-)-HRESIMS m / z 859.2313 [M-H] - (calcd for C 40 H 43 O 21 - , 859.2302).

[0060] Compound 2 Maltol [5”-O-6””'-(Kumatakenin B-4””-O-β-D-glucopyranosyl)-6'-O-(3”-hydroxy-3”-methylglutaryl)]-3-O-β-D-glucopyranoside

[0061] Pale yellow powder; ECD (MeOH) λ max (Δε) 215 (-12.44), 282 (-3.86) nm; UV (MeOH) λ max (logε) 205 (4.84), 255 (4.52), 314 (4.59) nm; IR ν max 3381, 2926, 1731, 1626, 1608, 1508, 1381, 1251, 1073, 845 cm -1 ; 1 H NMR and 13 C NMR are shown in Tables 1-1 and 1-2; (-)-HRESIMS m / z 829.2203 [M-H] - (calcd for C 39 H 41 O 20 - , 829.2197).

[0062] Compound 3 Maltol [5”-O-6””'-(Kumatakenin B-7”'-O-β-D-glucopyranosyl)-6'-O-(3”-hydroxy-3”-methylglutaryl)]-3-O-β-D-glucopyranoside

[0063] Pale yellow powder; ECD (MeOH) λ max (Δε) 203 (-8.51), 288 (-2.18) nm; UV (MeOH) λ max(logε) 205 (4.76), 230 (4.45), 255 (4.35), 309 (4.34), 327 (4.39) nm; IR ν max 3376, 2923, 1731, 1625, 1608, 1446, 1379, 1073 cm -1 ; 1 H NMR and 13 C NMR are shown in Tables 1-1 and 1-2; (-)-HRESIMS m / z 829.2204 [M-H] - (calcd for C 39 H 41 O 20 - , 829.2197).

[0064] Compound 4 Maltol [5”-O-6””'-(7”',4””-dihydroxy-8”'-methoxyisoflavone-4””-O-β-D-glucopyranosyl)-6'-O-(3”-hydroxy-3”-methylglutaryl)]-3-O-β-D-glucopyranoside

[0065] Pale yellow powder; ECD (MeOH) λ max (Δε) 231 (-7.06) nm; UV (MeOH) λ max (logε) 202 (4.79), 253 (4.74) nm; IR ν max 3381, 2925, 2854, 1731, 1644, 1608, 1510, 1451, 1253, 1074 cm -1 ; 1 H NMR and 13 C NMR are shown in Tables 1-1 and 1-2; (-)-HRESIMS m / z 859.2308 [M-H] - (calcd for C 40 H 43 O 21 - , 859.2302).

[0066] Compound 5 Maltol [5”-O-6””'-(Verbascoside-3””-O-β-D-glucopyranosyl)-6'-O-(3”-hydroxy-3”-methylglutaryl)]-3-O-β-D-glucopyranoside

[0067] Pale yellow powder; ECD (MeOH) λ max(Δε) 227 (-14.61), 283 (-3.07) nm; UV (MeOH) λ max (logε) 205 (4.72), 252 (4.66), 303 (4.19) nm; IR ν max 3390, 2924, 1731, 1644, 1623, 1515, 1457, 1265, 1199, 1073, 849 cm -1 ; 1 1H NMR and 13 13C NMR are shown in Tables 1-1 and 1-2; (-)-HRESIMS m / z 859.2308 [M-H] - (calcd for C 40 45 43 28 21 - , 859.2302).

[0068] Compound 6 Maltol [5”-O-6””'-(7”', 3””-dihydroxy-8”', 4””-dimethoxyisoflavone-3””-O-β-D-glucopyranosyl)-6'-O-(3”-hydroxy-3”-methylglutaryl)]-3-O-β-D-glucopyranoside

[0069] Pale yellow powder; ECD (MeOH) λ max (Δε) 224 (-12.71), 287 (-2.39) nm; UV (MeOH) λ max (logε) 205 (4.77), 255 (4.66) nm; IR ν max 3379, 2921, 2856, 1731, 1644, 1608, 1515, 1453, 1256, 1074 cm -1 ; 1 1H NMR and 13 13C NMR are shown in Tables 1-1 and 1-2; (-)-HRESIMS m / z 889.2415 [M-H] - (calcd for C 41 47 45 30 22 - , 889.2408).

[0070] Compound 8 (6”'S, 9”'S)-Maltol [5”-O-6””-asperuloside I-6'-O-(3”-hydroxy-3”-methylglutaryl)]-3-O-β-D-glucopyranoside

[0071] Pale yellow powder; ECD (MeOH) λ max (Δε) 242 (+19.83), 292 (-1.78) nm; UV (MeOH) λ max (logε) 242 (4.33) nm; IR ν max 3374, 2925, 2854, 1731, 1647, 1609, 1254, 1198, 1070 cm -1 ; 1 H NMR and 13 C NMR see Table 1 - 3; (-)-HRESIMS m / z 931.3440 [M - H] - (calcd for C 42 H 59 O 23 - , 931.3453).

[0072] Compound 9 (8”'R, 8””R)-maltol [5”-O-6””'-(secoisolariciresinol-4”'-O-β-D-glucopyranosyl)-6'-O-(3”-hydroxy-3”-methylglutaryl)]-3-O-β-D-glucopyranoside

[0073] Pale yellow powder; ECD (MeOH) λ max (Δε) 226 (-9.57), 273 (-4.03) nm; UV (MeOH) λ max (logε) 202 (5.08), 261 (4.17) nm; IR ν max 3361, 2920, 2852, 1732, 1645, 1602, 1515, 1258, 1073 cm -1 ; 1 H NMR and 13 C NMR see Table 1 - 4 and 1 - 5; (-)-HRESIMS m / z 937.3353 [M - H] - (calcd for C 44 H 57 O 22 - , 937.3347).

[0074] Compound 10 (7”'R, 8”'S)-maltol [5”-O-6””'-(dehydrodiconiferyl alcohol-4”'-O-β-D-glucopyranosyl)-6'-O-(3”-hydroxy-3”-methylglutaryl)]-3-O-β-D-glucopyranoside

[0075] Pale yellow powder; ECD (MeOH) λ max (Δε) 217 (-6.01), 281 (-6.33) nm; UV (MeOH) λ max (logε) 202 (4.77), 220 (4.52), 268 (4.22) nm; IR ν max 3381, 2924, 1731, 1645, 1603, 1258, 1073, 1032 cm -1 ; 1 1H NMR and 13 13C NMR are shown in Tables 1-4 and 1-5; (-)-HRESIMS m / z 979.3093 [M+HCOO] - (calcd for C 45 H 55 O 24 - , 979.3089).

[0076] Compound 11 (8”'R,7””S,8””R)-maltol [5”-O-6””'-(lariciresinol-4””-O-β-D-glucopyranosyl)-6'-O-(3”-hydroxy-3”-methylglutaryl)]-3-O-β-D-glucopyranoside

[0077] Pale yellow powder; ECD (MeOH) λ max (Δε) 220 (-27.28), 275 (-10.47) nm; UV (MeOH) λ max (logε) 206 (5.44), 259 (4.70) nm; IR ν max 3391, 2927, 1732, 1645, 1608, 1515, 1258, 1073, 1032 cm -1 ; 1 1H NMR and 13 13C NMR are shown in Tables 1-4 and 1-5; (+)-HRESIMS m / z 937.3334 [M+H] + (calcd for C 44 H 57 O 22 + , 937.3336).

[0078] Compound 12 (7”'R,8”'S,7””R,8””S)-maltol [5”-O-6””'-(pinoresinol-4”',4””-di-O-β-D-glucopyranosyl)-6'-O-(3”-hydroxy-3”-methylglutaryl)]-3-O-β-D-glucopyranoside

[0079] Pale yellow powder; ECD (MeOH) λ max (Δε) 203 (-10.50), 276 (-1.67) nm; UV (MeOH) λ max (logε) 202 (4.92), 228 (4.28), 261 (3.92) nm; IR ν max 3397, 2923, 1732, 1645, 1607, 1513, 1258, 1074 cm -1 ; 1 1H NMR and 13 13C NMR are shown in Tables 1-4 and 1-5; (+)-HRESIMS m / z 1097.3716 [M+H] + (calcd for C 50 50 65 26 27 + , 1097.3708).

[0080] Compound 13 maltol [5”-O-6””-(E)-coniferoside-6'-O-(3”-hydroxy-3”-methylglutaryl)]-3-O-β-D-glucopyranoside

[0081] Pale yellow powder; ECD (MeOH) λ max (Δε) 217 (-6.36), 261 (-2.63) nm; UV (MeOH) λ max (logε) 212 (4.42), 257 (4.31), 298 (3.62) nm; IR ν max 3381, 2922, 1732, 1645, 1608, 1512, 1256, 1199, 1073, 1023 cm -1 ; 1 1H NMR and 13 13C NMR are shown in Tables 1-4 and 1-5; (+)-HRESIMS m / z 779.2367 [M+Na] + (calcd for C 34 34 44 18 19 Na + , 779.2369).

[0082] Compound 14

[0083] White powder; 1 1H NMR and 13 13C NMR are shown in Table 1-6.

[0084] Compound 15

[0085] White powder; 1 1H NMR and 13 13C NMR are shown in Table 1-6.

[0086] Table 1-1 1H NMR data of Compounds 1-6 (CD3OD) 1

[0087]

[0088] At a 500 MHz, b 400 MHz. Measured; c,d Signals can be exchanged with each other

[0089] Table 1-2 13C NMR (126 MHz) data of Compounds 1-6 (CD3OD) 13

[0090]

[0091]

[0092] Table 1-3 1H NMR and 1 13C NMR data of Compound 8 (CD3OD) 13

[0093]

[0094]

[0095] 1 1H spectrum was measured at a 500 MHz; 13 13C spectrum was measured at b 126 MHz; c,d Signals can be interchanged; Assigned by e HMBC spectrum;

[0096] f Overlapped with the solvent peak, assigned by HSQC spectrum.

[0097] Table 1-4 1H NMR data of Compounds 9-13 (CD3OD) 1 ​​​​

[0098]

[0099]

[0100] At a 500 MHz, b 400 MHz, c measured at 600 MHz; d,g,h,I,k The signals are interchangeable; e Overlapping with the water peak

[0101] Table 1-5 13 13C NMR data of compounds 9-13 (CD3OD)

[0102]

[0103]

[0104] At a 101 MHz, b 126 MHz, c measured at 151 MHz; e-k,m,n,p-s The signals are interchangeable.

[0105] Table 1-6 1 1H NMR (700 Hz) and 13 13C NMR (151 Hz) data of compounds 14 and 15 (CD3OD)

[0106]

[0107]

[0108] Pharmacological experiments

[0109] Inhibitory effect of Example 1 on M2 macrophages

[0110] Mouse macrophages Ana-1 were cultured in 1640 medium with 10% FBS and seeded in 6-well plates. The test samples were pre-incubated with the cells at a concentration of 10 μM for 2 h, and Ana-1 cells were induced to polarize into M2 macrophages with 20 ng / mL of IL-4 and co-incubated for 24 h. The cells were harvested and centrifuged, and the cells were resuspended in 100 μL. Each group of cells was first blocked for non-specific binding sites with CD16 / 32, then APC-CD206 antibody was added, and incubated for 30 min under light protection. After washing with PBS, the proportion of M2 macrophages was measured by flow cytometry. The results (Table 2-1) showed that 5 compounds (1, 4, 5, 9, 11) had the effect of reducing the proportion of M2 macrophages at a concentration of 10.0 μM.

[0111] Table 2-1 Inhibitory effect of compound (10.0 μM) on M2 macrophages

[0112]

[0113]

[0114] **P < 0.01, compared with the model group

[0115] Effect Example 2 Dose-dependence of the inhibitory effect on M2 macrophage polarization

[0116] To study the dose-dependence of the active compound, the activities of 4 active compounds were further determined at different concentrations (1 μM, 2.5 μM, 10 μM). Mouse macrophages Ana-1 were cultured in 1640 medium with 10% FBS and seeded in 6-well plates. The test samples were pre-incubated with the cells at concentrations of 1, 2.5, and 10 μM for 2 h, and Ana-1 cells were induced to polarize into M2 type with 20 ng / mL of IL-4 and co-incubated for 24 h. The cells were collected and centrifuged, and the cells were resuspended in 100 μL. For each group of cells, non-specific binding sites were blocked with CD16 / 32 first, and then APC-CD206 antibody was added and incubated for 30 min under light protection. After washing with PBS, the proportion of M2 macrophages was measured by flow cytometry. The results are shown in Table 2-2.

[0117] Table 2-2 Inhibitory effect of compounds at different concentrations on M2 macrophages

[0118]

[0119] *P < 0.05, **P < 0.01, compared with the model group

[0120] Effect Example 3 Phagocytosis of tumor cells

[0121] Bone marrow cells were isolated from the hind leg femurs of mice and induced to be primary macrophages using CSF1. The macrophages were co-cultured with tumor cells labeled with CFSE dye. After treatment at 37 °C for 2 h, the macrophages were stained with APC-conjugated F4 / 80 and analyzed by flow cytometry. A total of 10,000 cells were analyzed in each sample. Unstained controls and single-stained cells were prepared for gating. The phagocytosis calculation method was the percentage of F4 / 80+CFSE+ cells among F4 / 80+ cells.

[0122] Table 2-3 Phagocytosis of tumor cells by macrophages

[0123]

[0124] *P < 0.05, **P < 0.01, compared with the control group

[0125] As can be seen from Effect Examples 1 and 2, the licoagroside B derivatives (1, 4, 5, 9, 11) obtained from Sophora flavescens Ait. showed varying degrees of inhibitory effects on M2 macrophages. Among them, Compound 1 had the strongest activity and had a significant reversal effect at 1.0 μM, but the dose-dependence was general; Compound 5 had a significant effect at 1.0 μM and showed dose-dependence at 2.5 μM and 10.0 μM; Compound 9 showed dose-dependence at 2.5 μM and 10.0 μM, but was ineffective at 1.0 μM; Compound 11 had a significant effect at 1.0 μM and showed dose-dependence at 1.0 μM and 2.5 μM.

[0126] As can be seen from Effect Example 3: Compounds (5, 9, 11) promoted the phagocytosis of macrophages at different concentrations. Among them, Compounds 5 and 11 had significant activities at a concentration of 2.0 μM.

[0127] The above indicates that such compounds can significantly reverse the polarization of M2 macrophages, thereby promoting macrophage phagocytosis, and have the potential to be developed into drugs for treating related diseases such as anti-tumor, anti-fibrotic diseases, anti-virus, anti-bacteria, and anti-infection.

Claims

1. A licoagroside B derivative of formula (I) or a pharmaceutically acceptable salt thereof, characterized in that, The parent nucleus of this class of compounds has the structure of general formula I, wherein R1 is independently selected from flavonoids and their glycosides, isoflavonoids and their glycosides, phenylpropanoids and their glycosides, lignans and their glycosides, ionol and their glycosides; R2 is independently selected from H, apiose; is independently selected from 2. The licoagroside B derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, wherein when R1 has a fragment of formula II, the parent nucleus I can be independently linked at the A1, A2 or A3 position; wherein A1, A2 and A3 are independently selected from H, OH, alkoxy of C1-C3 or monosaccharide; wherein when R1 has a fragment of formula III, the parent nucleus I can be independently linked at the B1, B2, B3 or B4 position of formula III; B1, B2, B3 and B4 are independently selected from H, OH, alkoxy of C1-C3 or monosaccharide; wherein when R1 has a fragment of formula IV, The mother nucleus I can be independently connected to the C1 or C2 position of Formula IV; C1 and C2 are independently selected from H, OH, alkoxy groups with 1 to 3 carbon atoms, or monosaccharides; independently selected from wherein when R1 has a fragment of formula V, The mother nucleus I can be independently linked to the D1, D2, D3 or D4 positions of formula V; D1, D2, D3 and D4 are independently selected from H, OH, alkoxy groups having 1 to 3 carbon atoms or monosaccharides; independently selected from wherein when R1 has a fragment of formula VI, The mother nucleus I can be independently connected to the E1, E2, E3 or E4 positions of formula VI; E1, E2, E3 and E4 are independently selected from H, OH, alkoxy groups having 1 to 3 carbon atoms or monosaccharides; independently selected from wherein when R1 has a fragment of formula VII, The mother nucleus I can be independently connected to the F1, F2, F3 or F4 positions of Formula VII; F1, F2, F3 and F4 are independently selected from H, OH, alkoxy groups having 1 to 3 carbon atoms or monosaccharides; independently selected from wherein when R1 has a fragment of formula VIII, The mother nucleus I can be independently linked to the positions G1, G2, G3 or G4 of formula VIII; G1, G2, G3 and G4 are independently selected from H, OH, alkoxy groups having 1 to 3 carbon atoms or monosaccharides; independently selected from wherein when R1 has a fragment of formula IX, the parent nucleus I can be independently linked at the K1, K2 or K3 position of formula IX; K1, K2 and K3 are independently selected from H, OH, alkoxy of C1-C3.

3. The licoagroside B derivative according to claims 1-2 or a pharmaceutically acceptable salt thereof, characterized in that, The alkoxy in A1, A2 and A3 is selected from OCH3 or OC2H5, and the monosaccharide is selected from glucose, xylose or apiose; the alkoxy in B1, B2, B3 and B4 is selected from OCH3 or OC2H5, and the monosaccharide is selected from glucose, xylose or apiose; the alkoxy in C1 and C2 is selected from OCH3 or OC2H5, and the monosaccharide is selected from glucose, xylose or apiose; the alkoxy in D1, D2, D3 and D4 is selected from OCH3 or OC2H5, and the monosaccharide is selected from glucose, xylose or apiose; the alkoxy in E1, E2, E3 and E4 is selected from OCH3 or OC2H5, and the monosaccharide is selected from glucose, xylose or apiose; the alkoxy in F1, F2, F3 and F4 is selected from OCH3 or OC2H5, and the monosaccharide is selected from glucose, xylose or apiose; the alkoxy in G1, G2, G3 and G4 is selected from OCH3 or OC2H5, and the monosaccharide is selected from glucose, xylose or apiose; the alkoxy in K1, K2 and K3 is selected from OCH3 or OC2H5, and the monosaccharide is selected from glucose, xylose or apiose.

4. The licoagroside B derivative or a pharmaceutically acceptable salt thereof according to claims 1-3, characterized in that, The described compound is selected from:

5. The licoagroside B derivative according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, characterized in that, The described pharmaceutically acceptable salt is selected from the salts formed by the reaction of the compound with inorganic bases and organic bases.

6. A method for preparing a licoagroside B derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, characterized in that, The described preparation method is as follows: After the Sophora subprostrata root medicinal material is extracted with water and / or ethanol, the extract is separated and purified by extraction and column chromatography to obtain the described compound.

7. A pharmaceutical composition, characterized in that, The described pharmaceutical composition comprises the licoagroside B derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1-5 and a pharmaceutically acceptable carrier or excipient.

8. The pharmaceutical composition according to claim 7, characterized in that, The dosage forms of the described pharmaceutical composition include tablets, capsules, pills, granules, powders, dripping pills, oral liquids or suspensions.

9. Use of the licoagroside B derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1-5 or the pharmaceutical composition according to any one of claims 7-8 in the preparation of a drug for treating diseases related to M2 macrophage polarization.

10. The application according to claim 9, wherein the M2 macrophage polarization-related diseases include tumor diseases, fibrosis diseases, and viral, bacterial, and other microbial infectious diseases.

Images