A pentacyclic triterpene derivative, its preparation method and medical use

Abstract

The application belongs to the technical field of medicine, and discloses a pentacyclic triterpene derivative, a preparation method and medical application of the pentacyclic triterpene derivative, which uses alpha-hederin or oleanolic acid as raw material, and utilizes esterification reaction or amidation reaction aiming at the carboxyl of C-28 to prepare the pentacyclic triterpene derivative. The pentacyclic triterpene derivative prepared by the application has the effects of anti-inflammation, anti-oxidation and anti-apoptosis, and the synthesis method is simple, the raw material is cheap and easy to obtain, and is suitable for industrial application.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and in particular relates to a pentacyclic triterpenoid derivative, its preparation method and pharmaceutical uses. Background Technology

[0002] Inflammation is a common and prevalent disease. A normal inflammatory response is a defense mechanism of the body, a biological reaction of the immune system to harmful stimuli (such as viruses, bacterial infections, toxins, toxic compounds, and tissue damage). Currently, the primary clinical treatment for inflammation is drug therapy. Common anti-inflammatory drugs are mainly divided into steroidal anti-inflammatory drugs (SAIDs) and non-steroidal anti-inflammatory drugs (NSAIDs), with NSAIDs being the most widely used anti-inflammatory drugs worldwide. Although they possess potent anti-inflammatory, analgesic, and antipyretic activities, they also have significant toxic side effects. Overall, safe and effective anti-inflammatory drugs remain extremely scarce in clinical practice, and the development of such drugs has enormous clinical need and significant social implications.

[0003] Pulsatilla saponin B5 is a zidonicane-type triterpenoid saponin extracted from the roots of Pulsatilla chinensis. It is also a pentacyclic triterpenoid compound with the aglycone structure of hederogenin. It is one of the important components of the extract of the traditional Chinese medicine Pulsatilla chinensis. Its structural formula is shown below. Figure 1 It has been reported that Pulsatilla saponin B5 possesses good anti-inflammatory activity. However, Pulsatilla saponin B5 is a pentasaccharide saponin with a molecular weight of 1221. It is extremely water-soluble (over 150 mg / mL in water at room temperature). Due to its low oral bioavailability (below 0.1%), large oral doses are required, resulting in insignificant anti-inflammatory activity in vivo. Although injection can improve the bioavailability of B5, its common properties as a triterpenoid saponin, such as cytotoxicity, hemolysis, and irritation, lead to the misconception that it cannot be administered by injection. Similarly, α-hederonine is also an important component extracted from the roots of Pulsatilla chinensis and can be obtained by hydrolyzing the ester bond at the C-28 position of Pulsatilla saponin B5. Unlike Pulsatilla saponin B5, its C-28 position is -COOH. Studies have found that α-hederine also has good anti-inflammatory activity, but its hemolytic toxicity still exists. According to previous structure-activity relationship analysis, the sugar chain at C-3 position is an essential group for anti-inflammatory activity, while the free carboxyl group at C-28 position is crucial for its cytotoxicity and hemolytic toxicity. Therefore, modifying its structure with the free carboxyl group at C-28 position as the target functional group has become a feasible approach. Summary of the Invention

[0004] The purpose of this invention is to solve the above problems by providing a pentacyclic triterpenoid derivative, its preparation method and pharmaceutical use. The pentacyclic triterpenoid derivative obtained by this invention has good anti-inflammatory, antioxidant and anti-apoptotic effects.

[0005] The technical solution of the present invention is as follows:

[0006] A pentacyclic triterpenoid derivative has the following general structural formula:

[0007] ,

[0008] Wherein, R1 is hydrogen, hydroxyl, C 1-8 The alkoxy group or -OX, where X is a monosaccharide, disaccharide, or trisaccharide; R2 is hydrogen, alkyl, hydroxyl, haloalkyl, or oxygen-substituted alkyl; R3 is -NHR4, -NR4, or -OR4, where R4 is hydrogen, alkyl, haloalkyl, ester, C 1-8 Alkoxy, aryl-C 1-6 Alkyl, aryloxy-C 1-6 Alkyl, hydroxy-C 1-6 Alkyl, amino-C 1-6 Alkyl, cycloalkyl-C 1-6 Alkyl, aromatic heterocyclic -C 1-6 Alkyl, aliphatic heterocycles -- C 1-6 Alkyl groups, amino acids, or amino acid esters.

[0009] Preferably, the glycosyl group in the monosaccharide, disaccharide, or trisaccharide is glucose, arabinose, rhamnose, galactose, xylose, or their acetylated form.

[0010] The pentacyclic triterpenoid derivatives are selected from the following structural formulas:

[0011] .

[0012] The present invention also provides the application of the aforementioned pentacyclic triterpenoid derivatives in the preparation of anti-inflammatory drugs, antioxidant drugs, or anti-apoptotic drugs.

[0013] Preferably, the anti-inflammatory drug is an anti-enteritis drug, an anti-pneumonia drug, an anti-myocarditis drug, an anti-pericarditis drug, an anti-hepatitis drug, or an anti-nephritis drug.

[0014] The present invention also provides a pharmaceutical composition comprising the aforementioned pentacyclic triterpenoid derivative or a pharmaceutically acceptable salt thereof.

[0015] The present invention also provides a pharmaceutical formulation comprising a therapeutically effective amount of the said pentacyclic triterpenoid derivative or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0016] Preferably, the pharmaceutical preparation is a topical preparation, an oral preparation, a rectal preparation, a subcutaneous injection preparation, an intramuscular injection preparation, or an intravenous injection preparation.

[0017] The present invention also provides a method for preparing the pentacyclic triterpenoid derivative, comprising the following steps: using α-hederone or oleanolic acid as starting materials, the pentacyclic triterpenoid derivative is prepared by esterification or amidation reaction.

[0018] Preferably, the esterification reaction includes the following steps: using α-hederone or oleanolic acid as starting materials, and in the presence of potassium carbonate and solvent, reacting with a halogenated compound to generate a pentacyclic triterpenoid derivative.

[0019] The molar ratio of α-hederone or oleanolic acid to the halogenated compound is 1:(1-2), preferably 1:1.5.

[0020] The molar amount of potassium carbonate is 2-4 times that of α-hederone or oleanolic acid, preferably 3 times.

[0021] Preferably, the halogenated compound is a halogenated aliphatic hydrocarbon, a halogenated cycloalkanes, a halogenated aromatic hydrocarbon, or a halogenated heterocyclic aromatic hydrocarbon. More preferably, the halogenated compound is 2-(bromomethyl)-3,5,6-trimethylpyrazine or (bromomethyl)cyclopropane.

[0022] Preferably, the amidation reaction includes the following steps: using α-hederone or oleanolic acid as starting materials, and in the presence of auxiliary reagents and solvents, reacting with amine compounds to generate pentacyclic triterpenoid derivatives.

[0023] The molar ratio of α-hederine or oleanolic acid to amine compounds is 1:(2-3), preferably 1:2.

[0024] Preferably, the auxiliary reagent is any one or more of 1-hydroxybenzotriazole, N,N-diisopropylethylamine, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, N,N'-carbonyldiimide, dicyclohexylcarbodiimide, diisopropylcarbodiimide, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, O-benzotriazole-tetramethylurea hexafluorophosphate, 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate, 2-succinimide-1,1,3,3-tetramethylurea tetrafluoroborate, N'-tetramethylurea tetrafluoroborate, diphenyl azidophosphate, 4-dimethylaminopyridine, and 1-hydroxy-7-azabenzotriazole.

[0025] Preferably, the auxiliary reagent is 1-hydroxybenzotriazole, N,N-diisopropylethylamine, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, wherein the molar amounts of the three are 2-3 times, 3-5 times, and 1.5-2 times that of α-hederone or oleanolic acid, respectively. More preferably, the molar amounts of 1-hydroxybenzotriazole, N,N-diisopropylethylamine, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride are 2 times, 3 times, and 1.5 times that of α-hederone, respectively, and the molar amounts of 1-hydroxybenzotriazole, N,N-diisopropylethylamine, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride are 2 times, 3 times, and 2 times that of oleanolic acid, respectively.

[0026] Preferably, the amine compound is NH2R4 or NHR4.

[0027] More preferably, the amine compound is any one of 3-aminomethylpyridine, 1-(4-chlorophenyl)ethylamine, cyclopropylmethylamine, 2-phenoxybenzylamine, (1-methyl-4-piperidin-)methylamine, 4-phenoxybenzylamine, n-butylamine, 6-methoxy-A-methyl-2-naphthylmethylamine, 1-(4-phenoxyphenyl)ethylamine, 2-cyclopropylethylamine hydrochloride, 2-aminomethylpyrazine, 3-aminomethyl-tetrahydrofuran, ethylamine hydrochloride, 2,2-difluoroethylamine, DL-alanine methyl ester hydrochloride, L-cyclopropylglycine, 4-amino-2-methyl-but-2-ol, 3-methoxypropylamine, isopentylamine, and 9-aminofluorene.

[0028] Preferably, in the above-mentioned esterification and amidation reactions, the solvent is a solvent commonly used in esterification or amidation reactions; more preferably, the solvent is N,N-dimethylformamide.

[0029] Preferably, in the above-mentioned pentacyclic triterpenoid derivative structural formula, R1 and R2 correspond to the corresponding substitution positions in α-hederone or oleanolic acid, and R3 or R4 corresponds to amine compounds or halogenated compounds.

[0030] The beneficial effects of this invention are as follows:

[0031] The pentacyclic triterpenoid derivatives obtained by this invention have anti-inflammatory, antioxidant and anti-apoptotic effects, and the synthesis method is simple, the raw materials are inexpensive and readily available, making them suitable for industrial application. Attached Figure Description

[0032] Figure 1 The chemical structural formula of Pulsatilla saponin B5;

[0033] Figure 2 The therapeutic effect of the pentacyclic triterpenoid derivative prepared in this invention on LPS-induced sepsis in mice. Detailed Implementation

[0034] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0035] Unless otherwise specified, all reagents involved in the embodiments of this invention are commercially available products and can be purchased through commercial channels.

[0036] The Chinese names corresponding to the English abbreviations involved in this invention are as follows: HoBt: 1-hydroxybenzotriazole; DIPEA: N,N-diisopropylethylamine; DMF: N,N-dimethylformamide; EDCI: 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride; TLC: thin-layer chromatography.

[0037] I. Synthesis of Compounds

[0038] The structural formulas of the compounds synthesized in this invention are as follows:

[0039] .

[0040] Example 1 (Preparation of compound HSC-1)

[0041] α-Hydroxylin and eosin (100 mg, 0.13 mmol), HoBt (36 mg, 0.26 mmol), and DIPEA (52 mg, 0.39 mmol) were dissolved in DMF (2 mL). After stirring at room temperature for 5 minutes, EDCI (51 mg, 0.2 mmol) was added and stirred for 30 minutes. Then, 3-aminomethylpyridine (28.8 mg, 0.26 mmol) was added and the reaction was carried out overnight. The reaction was detected by TLC. After the reaction was completed, the reaction solution was poured into 15 mL of ice water, extracted twice with ethyl acetate (40 mL), the organic layer was washed twice with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure to obtain the crude product. Preparative liquid chromatography (methanol:water = 75:25) yielded 74 mg of a white solid powder, with a yield of 66%.

[0042] 1 H NMR (600 MHz, Methanol- d 4) δ 8.51 (d, J = 2.2 Hz, 1H), 8.41 (dd, J = 4.9, 1.6 Hz, 1H), 7.80 (dt, J = 7.9, 2.0 Hz, 1H), 7.39 (dd, J= 7.9, 4.8Hz, 1H), 5.33 (t, J = 3.7 Hz, 1H, H12), 5.16 (d, J = 1.8 Hz, 1H, H-1 of rha),4.56 (d, J = 5.3 Hz, 1H, H-1 of ara), 1.26 (s, 3H, H-6 of rha), 1.17 (s, 3H), 0.96 (s, 3H), 0.93 (s, 3H), 0.92 (s, 3H), 0.69 (s, 3H), 0.51 (s, 3H).

[0043] 13 CNMR (151 MHz, MeOD) δ 180.36, 149.82, 148.57, 145.09, 138.07,137.13, 125.11, 123.98, 104.32, 101.88, 82.19, 76.64, 73.92, 73.67, 72.13,72.01, 70.16, 69.14, 64.78, 64.55, 48.04, 47.56, 43.91, 42.94, 42.56, 41.76,40.55, 39.61, 37.56, 35.07, 34.48, 33.55, 33.24, 31.60, 28.48, 26.50 (×2),24.50, 24.02, 23.97, 20.86, 18.73, 17.97, 17.79, 16.36, 14.46, 13.71. HRMS (m / z) [M + H] + calcd for C 47 H 72 N2O 11 : 841.5208; found: 841.5208.

[0044] Example 2 (Preparation of compound HSC-2)

[0045] The preparation method of compound HSC-2 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with 1-(4-chlorophenyl)ethylamine (41.44 mg, 0.26 mmol), and 79 mg of white solid powder is obtained, with a yield of 67%.

[0046] 1HNMR (600 MHz, Methanol-d4) δ 7.32 (d, J = 4.4 Hz, 2H), 7.30 (d, J =2.2 Hz, 2H), 5.17 (s, 1H, H-1 of rha), 4.56 (s, 1H, H-1 of ara), 1.26 (s, 3H, -NHCH(CH3)-Ar-Cl), 1.25 (s, 3H, H-6 of rha), 1.17 (s, 3H), 1.16 (s, 3H), 0.96 (s, 3H), 0.96 (s, 3H), 0.92 (s, 3H), 0.89 (s, 3H).

[0047] 13 CNMR (151 MHz, MeOD) δ 179.14, 145.26, 144.61, 133.85, 129.57,129.45, 129.26, 129.01, 123.96, 104.32, 101.87, 82.21, 76.64, 73.92, 73.66,72.13, 72.00, 70.15, 69.13, 64.77, 64.56, 61.53, 49.87, 48.07, 48.05, 47.38,43.91, 42.95, 42.49, 40.52, 39.65, 37.53, 33.62, 31.57, 28.48, 26.50, 26.41,24.08 (×2), 23.97, 23.74, 22.38, 21.90, 18.72, 17.97, 17.93, 17.85, 16.39,13.72. HRMS (m / z) [M + H] + calcd for C 49 H 74 ClNO11: 888.5023; found: 888.5021.

[0048] Example 3 (Preparation of compound HSC-3)

[0049] α-Hydroxylin and eosin (100 mg, 0.13 mmol) were dissolved in DMF (2.5 mL), potassium carbonate (55 mg, 0.39 mmol) was added, followed by 2-(bromomethyl)-3,5,6-trimethylpyrazine (43 mg, 0.195 mmol). The reaction was carried out at 50 °C for 12 hours. After the reaction was completed, the reaction solution was poured into 15 mL of ice water, extracted twice with ethyl acetate (40 mL), the organic layer was washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure to obtain the crude product. Purification by preparative liquid chromatography (methanol:water = 75:25) yielded 75 mg of a white solid powder, with a yield of 64%.

[0050] 1 HNMR (600 MHz, Methanol-d4) δ 5.16 (s, 1H, H-1 of rha) 4.56 (d, J =5.2 Hz, 1H, H-1 of ara) 2.59 (s, 3H), 2.52 (s, 3H), 2.51 (s, 3H), 1.26 (s, 3H, H-6 of rha) 1.24 (s, 3H), 1.14 (s, 3H), 0.95 (s, 3H), 0.93 (s, 3H), 0.91 (s, 3H), 0.71 (s, 3H).

[0051] 13 C NMR (151 MHz, DMSO) δ 176.10, 150.63, 148.70, 148.25, 145.01,143.24, 121.95, 102.98, 99.93, 79.26, 74.19, 72.89, 71.98, 70.39, 70.32,68.11, 67.84, 64.46, 64.24, 59.77, 46.98, 46.17, 46.13, 45.39, 42.23, 41.20,40.95, 40.06, 38.68, 38.16, 35.90, 33.17, 32.71, 32.03, 30.35, 26.98, 25.54,25.28, 23.34, 21.21, 20.94, 20.78, 20.08, 17.77, 17.00, 16.34, 15.49, 14.10,13.03. HRMS (m / z) [M + H] + calcd for C 49 H 76 N2O12 : 885.5471; found: 885.5476.

[0052] Example 4 (Preparation of compound HSC-4)

[0053] The preparation method of compound HSC-4 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with cyclopropylmethylamine (19 mg, 0.26 mmol), and 76 mg of white solid powder is obtained, with a yield of 71%.

[0054] 1 H NMR (600 MHz, Methanol-d4) δ5.37 (t, J = 3.7 Hz, 1H, H12) 5.15 (s,1H, H-1 of rha), 4.55 (d, J = 5.3 Hz, 1H, H-1 of ara) 3.13 – 2.86 (m, 2H), 1.23 (s, 3H, H-6 of rha), 1.24 – 1.23 (m, 4H) 1.19 (s, 3H), 0.97 (s, 3H), 0.95 (s, 3H), 0.91 (s, 3H), 0.81 (s, 3H), 0.69 (s, 3H), 0.45 (d, J = 8.2 Hz, 2H), 0.22– 0.15 (m, 2H).

[0055] 13 C NMR (151 MHz, DMSO) δ 176.16, 144.16, 121.48, 102.98, 99.93,79.30, 74.19, 72.93, 72.05, 70.38, 68.13, 67.89, 64.46, 62.48, 59.78, 47.14,46.20, 46.08, 45.23, 43.25, 42.26, 41.36, 40.54, 40.06, 38.85, 38.23, 35.97,33.68, 32.95, 32.80, 32.00, 30.45, 26.96, 25.64, 25.32, 23.58, 22.96, 22.36,17.79, 17.10, 16.94, 15.57, 14.11, 13.03, 11.02. HRMS (m / z) [M + H] +calcd forC 45 H 73 NO 11 : 804.5256; found:804.5262.

[0056] Example 5 (Preparation of compound HSC-5)

[0057] The preparation method of compound HSC-5 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with 2-phenoxybenzylamine (62.7 mg, 0.26 mmol), and 85.6 mg of white solid powder is obtained, with a yield of 69%.

[0058] 1 H NMR (600 MHz, Methanol- d 4) δ 8.05 (dd, 1H, -CONHC-), 7.65 (dd, J =8.4, 6.8 Hz, 2H), 7.52 (d, J = 8.1 Hz, 3H), 7.37 (t, J = 1.8 Hz, 1H), 7.27(td, 1H), 7.12 (dd, 1H), 7.00 (dd, 1H), 4.58 (d, J = 5.2 Hz, 1H, H-1 of ara)1.27 (s, 3H), 1.26 (s, 3H), 1.03 (s, 3H), 0.99 (s, 3H), 0.98 (s, 3H), 0.86 (s, 3H), 0.72 (s, 3H).

[0059] 13C NMR (151 MHz, MeOD) δ 174.96, 158.65, 156.46, 144.58, 131.16,131.07, 130.22, 130.00, 126.47, 125.13, 124.84, 124.53, 120.85, 119.71,119.42, 104.34, 101.90, 82.19, 76.68, 73.93, 73.67, 72.15, 72.02, 70.18,69.14, 64.78, 64.58, 48.11, 46.70, 43.94, 43.18, 43.09, 40.74, 39.69, 37.61,37.52, 34.57, 33.67, 33.46, 33.30, 31.57, 31.53, 29.14, 26.51, 26.30, 24.57,24.16, 24.04, 23.87, 18.79, 18.13, 17.98, 16.44, 13.74. HRMS (m / z) [M + H] + calcd for C 54 H 77 NO 12 : 932.5518; found: 932.5521.

[0060] Example 6 (Preparation of compound HSC-6)

[0061] The preparation method of compound HSC-6 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with (1-methyl-4-piperidin-)methylamine (34.1 mg, 0.26 mmol), and 71 mg of white solid powder is obtained, with a yield of 62%.

[0062] 1H NMR (600 MHz, Methanol-d4) δ 5.36 (t, J = 3.7 Hz, 1H, H12), 5.16 (d, J = 1.7 Hz, 1H, H-1 of rha), 4.56 (d, J = 5.2 Hz, 1H, H-1 of ara), 3.03(ddd, J = 68.2, 13.2, 6.9 Hz, 2H,), 2.26 (s, 3H), 1.25 (d, J = 6.2 Hz, 3H, H-6of rha), 1.20 (s, 3H), 0.98 (s, 3H), 0.96 (s, 3H), 0.92 (s, 3H), 0.79 (s,3H), 0.71 (s, 3H).

[0063] 13 C-NMR (151 MHz, MeOD) δ 180.34, 145.45, 123.90, 104.33, 101.88,82.18, 76.65, 73.92, 73.67, 72.14, 72.01, 70.16, 69.15, 64.79, 64.56, 56.36,49.85, 48.92, 48.07, 47.63, 47.58, 46.36,46.04 43.93, 43.05, 42.71, 40.66,39.63, 37.61, 36.31, 35.12, 34.53, HRMS (m / z) [M + H] + calcd for C 48 H 80 N2O 11 : 861.5834; found: 861.5810.

[0064] Example 7 (Preparation of compound HSC-7)

[0065] The preparation method of compound HSC-7 is the same as that of compound HSC-1. 3-Aminomethylpyridine (28.8 mg, 0.26 mmol) is converted to 4-phenoxybenzylamine (53 mg, 0.26 mmol) to obtain 84 mg of white solid powder, with a yield of 68%.

[0066] 1 H NMR (600 MHz, MeOD) δ 7.24 (t, J = 7.7 Hz, 2H), 7.20 (d, J = 8.2Hz, 2H), 7.01 (t, J = 7.4 Hz, 1H), 6.87 (d, J = 8.0 Hz, 2H), 6.81 (d, J = 8.2Hz, 2H), 4.46 (d, J = 5.3 Hz, 1H, H-1 of ara) 5.10 (s, 1H H-1 of rha) , 1.17(s, 3H, H-6 of rha), 1.08 (s, 3H), 0.87 (s, 3H), 0.84 (s, 3H), 0.83 (s, 3H),0.61 (s, 3H), 0.45 (s, 3H).

[0067] 13 CNMR (600 MHz, MeOD) δ 179.98, 158.68, 157.63, 145.20, 135.44, 130.83(×2), 130.63(×2), 124.30, 123.87, 119.69(×2), 119.67 (×2), 104.38,101.81, 82.14, 76.54, 73.84, 73.71, 72.05, 71.92, 70.10, 69.21, 64.91, 64.47,47.99, 47.57, 47.45, 47.43, 43.88, 43.81, 43.68, 42.92, 42.54, 40.50, 39.57,37.52, 35.10, 34.35, 33.62, 33.20, 31.60 (×2), 28.41, 26.50, 24.51, 24.06,18.77, 18.00, 17.94, 16.51, 13.77. HRMS (m / z) [M + H] + calcd for C 54 H 77 NO 12 :932.5518; found: 932.5518.

[0068] Example 8 (Preparation of compound HSC-8)

[0069] The preparation method of compound HSC-8 is the same as that of compound HSC-1. 3-Aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with n-butylamine (19.5 mg, 0.26 mmol) to obtain 70 mg of white solid powder, with a yield of 66%.

[0070] 1 H NMR (600 MHz, Methanol- d 4) δ 5.35 (t, J = 3.8 Hz, 1H, H12), 5.15 (d, J = 1.8 Hz, 1H, H-1 of rha), 4.55 (d, J = 5.2 Hz, 1H, H-1 of ara), 3.24 – 3.02 (m, 2H, -NHCH2CH2CH2CH3), 1.48 – 1.43 (m, 2H), 1.36 – 1.31 (m, 2H), 1.24 (dd, J =6.7, 3.5 Hz, 3H, H-6 of rha), 1.19 (s, 3H), 0.97 (s, 3H), 0.94 (s, 3H), 0.92(s, 3H), 0.91 (s, 3H), 0.79 (s, 3H), 0.70 (s, 3H).

[0071] 13 C NMR (151 MHz, MeOD) δ 180.27, 145.46, 123.96, 104.34, 101.89,82.20, 76.66, 73.93, 73.68, 72.14, 72.02, 70.16, 69.15, 64.79, 64.56, 48.95,48.07, 47.68, 47.51, 43.93, 43.05, 42.68, 40.64, 40.58, 40.45, 39.64, 37.60,35.14, 34.32, 33.57, 33.28, 32.61, 31.61, 28.51, 26.49, 24.57, 24.01(×2),21.34, 18.77, 17.97, 17.96, 16.38, 14.19, 13.72. HRMS (m / z) [M + H] + calcd forC 45 H 75 NO11 : 806.5412; found: 806.5397.

[0072] Example 9 (Preparation of compound HSC-9)

[0073] The preparation method of compound HSC-9 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with 6-methoxy-A-methyl-2-naphthylamine (53.6 mg, 0.26 mmol), and 77 mg of white solid powder is obtained, with a yield of 62%.

[0074] 1 H NMR (600 MHz, DMSO- d 6): δ7.76 – 7.65 (m, 3H), 7.43 (d, J = 8.5 Hz,1H), 7.26 (s, 1H), 7.13 (d, J = 9.0, 2.3 Hz, 1H), 4.30 (d, J = 6.2 Hz, 1H, H-1of ara), 3.84 (s, 3H, -COCH3), 1.42 (d, J = 5.6 Hz, 3H,-NHCH(CH3)C-), 1.05 (s,3H, H-6 of rha), 0.86 (s, 6H), 1.07 (s, 3H), 0.91 (s, 3H), 0.87 (s, 3H), 0.52(s, 3H) .

[0075] 13C NMR (151 MHz, DMSO) δ 175.18, 156.97, 144.20, 140.38, 139.96,133.22, 129.09, 128.28, 126.60, 125.68, 124.12, 118.59, 118.53, 105.75,103.11, 79.31, 74.18, 74.12, 73.05, 71.99, 70.39, 70.35, 68.14, 67.97, 64.66,62.39, 55.19, 47.77, 47.53, 47.17, 46.19, 46.15, 45.20, 45.19, 42.25, 41.28,40.67, 38.82, 38.70, 35.93, 33.60, 33.10, 33.02, 32.57, 30.54, 26.99, 25.74,25.37, 23.78, 22.25, 16.83, 16.60, 15.39, 13.10. HRMS (m / z) [M + H] + calcd forC 54 H 79 NO 12 : 934.5675; found: 934.5637.

[0076] Example 10 (Preparation of compound HSC-10)

[0077] The preparation method of compound HSC-10 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with 1-(4-phenoxyphenyl)ethylamine (56.8 mg, 0.26 mmol), and 80 mg of white solid powder is obtained, with a yield of 64%.

[0078] 1 H NMR (600 MHz, MeOD) δ7.28 – 7.20 (m, 4H), 7.01 (q, J = 7.2 Hz, 1H), 6.88 (d, J = 8.1 Hz, 2H), 6.83 (t, J = 9.2 Hz, 2H), 5.09 (d, J = 2.9 Hz, 1H), 4.45 (d, J = 5.3 Hz, 1H, H-1 of ara) 1.37 (d, J= 6.9 Hz, 3H,-NHC(CH3)C-), 1.08 (s, 3H), 1.07 (s, 3H), 0.87 (s, 3H), 0.86(s, 3H), 0.83 (s, 3H),0.79 (s, 3H), 0.61 (s, 3H).

[0079] 13 C NMR (151 MHz, MeOD) δ 179.08, 158.75, 157.76, 145.31, 140.89, 130.86(×2), 130.84(×2), 124.40, 124.29, 119.83(×2), 119.72(×2), 104.42,101.86, 82.14, 76.58, 73.88, 72.07, 71.96, 70.13, 69.28, 69.25, 64.93, 64.47,49.84, 48.00, 47.61, 47.39, 47.21, 43.90, 43.00, 42.97, 40.52, 39.63, 37.52,35.08, 34.14, 33.33, 28.45, 28.42, 26.51(×2), 26.41, 26.38, 24.07, 22.63,22.23, 18.70, 18.04, 17.98, 16.60, 16.44, 13.75. HRMS (m / z) [M + H] + calcd forC55H79NO12: 946.5675; found: 946.5633.

[0080] Example 11 (Preparation of compound HSC-11)

[0081] The preparation method of compound HSC-11 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with 2-cyclopropylethylamine hydrochloride (32.4 mg, 0.26 mmol), and 74 mg of white solid powder is obtained, with a yield of 68%.

[0082] 1 HNMR (600 MHz, Methanol- d 4) δ 5.41 – 5.32 (m, 1H,H12), 5.17 (d, J =1.8 Hz, 1H, H-1 of rha), 4.57 (d, J= 5.2 Hz, 1H, H-1 of ara),3.32 (dt, J =3.4, 1.7 Hz, 2H), 1.25 (d, J = 6.3 Hz, 3H), 1.20 (s, 3H), 0.99 (s, 3H), 0.96(s, 3H), 0.92 (s, 3H), 0.80 (s, 3H), 0.72 (s, 3H), 0.50 – 0.43 (m, 2H), 0.11 –0.05 (m, 2H).

[0083] 13 C NMR (151 MHz, MeOD) δ 180.12, 145.48, 124.01, 104.32, 101.88,82.19, 76.65, 73.92, 73.67, 72.13, 72.00, 70.15, 69.14, 64.78, 64.55, 48.93,48.06, 47.70, 47.44, 43.92, 43.03, 42.69, 40.88, 40.63, 39.63, 37.59, 35.40,35.12, 34.16, 33.56, 33.21, 31.61, 28.48, 26.51, 24.56, 24.08, 24.00, 18.76,17.96, 17.91, 16.39, 13.71, 9.61, 4.82, 4.71. HRMS (m / z) [M + H] + calcd forC 46 H 75 NO 11 : 818.5412; found: 818.5418.

[0084] Example 12 (Preparation of compound HSC-12)

[0085] The preparation method of compound HSC-12 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with 2-aminomethylpyrazine (29 mg, 0.26 mmol), and 70 mg of white solid powder is obtained, with a yield of 63%.

[0086] 1 HNMR (600 MHz, Methanol- d 4) δ 8.60 (d, J= 1.5 Hz, 1H), 8.57 (dd, J =2.6, 1.5 Hz, 1H), 8.49 (d, J = 2.6 Hz, 1H), 5.38 (t, J = 3.7 Hz, 1H, H12), 5.16 (d, J = 1.8 Hz, 1H, H1 of rha), 4.56 (d, J = 5.4 Hz, 1H, H1 of ara), 1.25(d, J = 6.2 Hz, 3H, H-6 of rha), 1.19 (s, 3H), 0.97 (s, 3H), 0.93 (s, 3H), 0.92 (s, 3H), 0.69 (s, 3H), 0.54 (s, 3H).

[0087] 13 C NMR (600 MHz, DMSO) δ 176.74, 154.66, 143.87, 143.71, 143.55,142.92, 121.70, 102.96, 99.94, 79.28, 74.22, 72.83, 71.98, 70.39, 70.32,68.14, 67.80, 64.41, 62.43, 47.09, 46.16, 45.95, 45.38, 42.60, 42.24, 41.26,40.62, 38.79, 38.18, 35.93, 33.58, 32.91, 32.70, 31.90, 30.46, 26.93, 25.65,25.30, 23.57, 22.89, 22.35, 17.78, 17.06, 16.50, 15.53, 13.02. HRMS (m / z) [M +H] + calcd for C 46 H 71 N3O 11 : 842.5161; found: 842.5160.

[0088] Example 13 (Preparation of compound HSC-13)

[0089] The preparation method of compound HSC-13 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with 3-aminomethyl-tetrahydrofuran (27 mg, 0.26 mmol), and 69 mg of white solid powder is obtained, with a yield of 63%.

[0090] 1 HNMR (600 MHz, Methanol- d 4) δ 7.41 (q, J = 5.5 Hz, 1H, -CONH-), 5.37(q, J = 3.7 Hz, 1H,H12), 5.17 (d, J = 1.7 Hz, 1H, H1 of rha), 4.57 (d, J =5.2 Hz, 1H, H1 of ara), 2.50 (dt, J = 7.5, 3.7 Hz, 1H,H-3 ofTetrahydrofuran-3-yl)methanamine), 1.26 (d, J = 6.2 Hz, 3H), 1.21 (s, 3H), 0.99 (s, 3H), 0.97 (s, 3H), 0.93 (s, 3H), 0.80 (s, 3H), 0.72 (s, 3H).

[0091] 13 CNMR (151 MHz, MeOD) δ 180.49, 145.34, 123.96, 104.32, 101.86,82.19, 76.64, 73.91, 73.66, 72.42, 72.31, 72.12, 71.99, 70.14, 69.13, 68.62,64.55, 48.92, 48.06, 47.59, 43.91, 43.04, 40.61, 40.38, 40.32, 39.62, 37.60,35.11, 34.47, 33.58, 33.32, 31.60, 30.91, 30.86, 28.53, 26.50(×2), 24.56,24.03(×2), 23.94, 18.77, 18.05, 17.97, 16.39, 13.73. HRMS(m / z)[M + H] + calcdfor C 46 H75 NO 12 : 834.5362; found: 834.5364.

[0092] Example 14 (Preparation of compound HSC-14)

[0093] The preparation method of compound HSC-14 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with ethylamine hydrochloride (21.7 mg, 0.26 mmol), and 75 mg of white solid powder is obtained, with a yield of 73%.

[0094] 1 H-NMR (600 MHz, Methanol- d 4) δ 5.37 (t, J = 3.7 Hz, 1H, H12), 5.16 (d, J = 1.7 Hz, 1H, H1 of rha), 4.56 (d, J = 5.2 Hz, 1H, H1 of ara), 3.32 (p, J = 1.7Hz, 2H), 1.25 (d, J = 6.2 Hz, 3H, H-6 of rha), 1.20 (s, 3H), 1.09 (t, J = 7.2Hz, 3H,-NHCH2CH3), 0.99 (s, 3H), 0.96 (s, 3H), 0.92 (s, 3H), 0.80 (s, 3H), 0.71 (s, 3H).

[0095] 13C NMR (151 MHz, DMSO) δ 175.98, 144.14, 121.45, 102.99, 99.92,79.29, 74.19, 72.91, 72.01, 70.43, 70.34, 68.12, 67.86, 64.47, 62.44, 47.13,46.19, 46.08, 45.08, 42.26, 41.29, 40.43, 38.21, 35.98, 33.66, 33.58, 33.46,32.96, 32.76, 31.94, 30.46, 26.95, 25.67, 25.32, 23.59, 22.93, 22.29, 17.79,17.08, 16.90, 15.56, 14.88, 13.05. HRMS (m / z) [M + H] + calcd for C 43 H 71 NO 11 :778.5099; found: 778.5110.

[0096] Example 15 (Preparation of compound HSC-15)

[0097] The preparation method of compound HSC-15 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with 2,2-difluoroethylamine (21.5 mg, 0.26 mmol), and 75 mg of white solid powder is obtained, with a yield of 70%.

[0098] 1 HNMR (600 MHz, Methanol- d 4) δ 5.83 (tt, J = 56.7, 4.4 Hz, 1H), 5.33(t, J = 3.7 Hz, 1H, H12), 5.15 (d, J = 1.8 Hz, 1H, H1 of rha), 4.55 (d, J = 5.2Hz, 1H, H1 of ara), 3.90 (dd, J = 3.5, 1.7 Hz, 2H), 1.24 (d, J= 6.2 Hz, 3H, H-6 of rha), 1.19 (s, 3H), 0.97 (s, 3H), 0.94 (s, 3H), 0.91 (s, 3H), 0.77 (s,3H), 0.70 (s, 3H).

[0099] 13 C NMR (151 MHz, DMSO) δ 177.43, 143.83, 121.66, 114.42, 103.01,99.96, 79.34, 74.22, 72.94, 72.06, 70.47, 70.39, 68.15, 67.90, 64.48, 62.49,47.14, 46.24, 45.93, 45.50, 42.29, 41.31, 40.41, 38.84, 38.25, 36.00, 35.85,33.57, 32.91, 32.67, 32.00, 30.42, 26.91, 25.65, 25.34, 23.52, 22.95, 22.21,17.81, 17.10, 16.64, 15.60, 13.06. HRMS (m / z) [M + H] + calcd for C 43 H 69 F2NO 11 :814.4911; found: 814.4920.

[0100] Example 16 (Preparation of compound HSC-16)

[0101] The preparation method of compound HSC-16 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with DL-alanine methyl ester hydrochloride (37 mg, 0.26 mmol), and 80 mg of white solid powder is obtained, with a yield of 72%.

[0102] 1 H NMR (600 MHz, DMSO- d 6) δ 7.52 (d, J = 7.2 Hz, 1H), 5.19 (t, J = 3.9Hz, 1H, H12) ,5.06 (s, 1H, H1 of rha) , 4.32 (d, J= 6.1 Hz, 1H, H1 of ara) ,3.57 (s, 3H), 1.24 (s, 3H) 1.24 (d, J = 3.4 Hz, 3H, H-6 of rha), 1.09 (s, 3H), 0.88 (s, 3H), 0.86 (s, 6H), 0.65 (s, 3H), 0.56 (s, 3H).

[0103] 13 C NMR (600 MHz, DMSO) δ 176.21, 173.31, 144.12, 121.33, 102.98,99.93, 79.30, 74.19, 72.93, 72.04, 72.01, 70.46, 70.37, 68.13, 64.46, 62.48,51.71, 51.69, 47.45, 47.15, 46.20, 45.11, 42.26, 41.34, 40.31, 38.87, 38.24,35.97, 32.92, 32.29, 31.97, 30.41, 26.79, 25.60, 25.32, 23.58(×2), 23.56,22.95, 17.78, 16.89, 16.77, 16.66, 15.59, 13.03. HRMS (m / z) [M + H] + calcd forC 45 H 73 NO 13 : 836.5154; found: 836.5152.

[0104] Example 17 (Preparation of compound HSC-17)

[0105] The preparation method of compound HSC-17 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with L-cyclopropylglycine (30.6 mg, 0.26 mmol), and 79 mg of white solid powder is obtained, with a yield of 69%.

[0106] 1 H NMR (600 MHz, DMSO- d 6) δ 8.14 (d, J = 8.4 Hz, 1H), 5.30 (t, J = 3.8Hz, 1H, H12), 5.07 (d, J= 1.7 Hz, 1H, H1 of rha), 4.34 (d, J = 6.1 Hz, 1H, H1of ara), 1.19 (s, 3H), 1.08 (d, J = 6.2 Hz, 3H), 0.95 (s, 6H), 0.90 – 0.84 (m, 6H), 0.74 (s, 3H), 0.58 (s, 3H).

[0107] 13 C NMR (151 MHz, DMSO) δ 178.59, 173.55, 142.81, 128.18, 102.99,99.94, 79.27, 74.20, 72.88, 72.00, 70.40, 70.33, 68.14, 67.83, 64.44, 62.44,47.15, 47.00, 46.19, 45.46, 44.89, 42.27, 41.52, 41.23, 40.06, 38.96, 38.23,35.96, 32.90, 32.84, 32.51, 31.84, 30.32, 27.53, 25.41(×2), 25.30, 23.32(×2), 22.96, 22.60, 20.79, 17.80, 17.07, 16.92, 15.58, 13.04. HRMS (m / z) [M + H] + calcd for C 47 H 75 NO 13 : 862.5311; found: 862.5310.

[0108] Example 18 (Preparation of compound HSC-18)

[0109] The preparation method of compound HSC-18 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with 4-amino-2-methyl-but-2-ol (27.5 mg, 0.26 mmol), and 66 mg of white solid powder is obtained, with a yield of 60%.

[0110] 1 H NMR (600 MHz, DMSO- d 6) δ 7.23 (t, J= 5.3 Hz, 1H), 5.24 – 5.17 (m,1H, H12), 5.06 (d, J = 1.6 Hz, 1H, H1 of rha), 4.32 (d, J = 6.2 Hz, 1H, H1 ofara), 3.36 – 3.26 (m, 2H), 1.80 (t, J = 9.0, 3.7 Hz, 2H), 1.08 (d, J = 2.0 Hz, 6H), 1.07 (s, 6H), 0.86 (d, J = 3.5 Hz, 9H), 0.67 (s, 3H), 0.56 (s, 3H).

[0111] 13 C NMR (151 MHz, DMSO) δ 175.92, 143.93, 121.68, 102.98, 99.92,79.29, 74.18, 72.89, 72.00, 70.40, 70.34, 68.56, 68.12, 67.84, 64.45, 62.44,47.13, 46.18, 46.08, 45.12, 42.26, 42.02, 41.28, 40.61, 40.06, 38.91, 38.21,35.97, 35.43, 33.65, 32.94, 31.92, 30.47, 29.52(×2), 29.15(×2), 26.95,25.67, 25.31, 23.56, 17.79, 17.09, 16.85, 15.57, 13.03. HRMS(m / z)[M + H] + calcd for C 46 H 77 NO 12 : 836.5518; found: 836.5522.

[0112] Example 19 (Preparation of compound HSC-19)

[0113] The preparation method of compound HSC-19 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with 3-methoxypropylamine (24 mg, 0.26 mmol), and 70 mg of white solid powder is obtained, with a yield of 64%.

[0114] 1 H NMR (600 MHz, DMSO- d 6) δ 7.21 (t, J = 5.6 Hz, 1H), 5.21 (d, J = 3.7Hz, 1H, H12), 5.06 (d, J = 1.6 Hz, 1H, H1 of rha), 4.32 (d, J = 6.1 Hz, 1H, H1of ara), 3.20 (s, 3H), 3.12 – 3.05 (m, 2H), 1.59 (t, J = 6.6 Hz, 2H), 1.09 (s, 3H), 1.07 (d, J = 6.2 Hz, 3H), 0.87 (s, 3H), 0.86 (s, 6H), 0.66 (s, 3H), 0.56 (s, 3H).

[0115] 13 C NMR (151 MHz, DMSO) δ 176.19, 144.12, 121.49, 102.99, 99.93,79.30, 74.18, 72.89, 72.00, 70.40, 70.34, 70.22, 68.12, 67.85, 64.46, 59.78,57.94, 47.12, 46.19, 46.06, 45.22, 42.25, 41.30, 40.52, 38.88, 38.21, 36.41,35.97, 33.64, 32.95, 31.95, 30.46, 29.13, 26.95, 25.67(×2), 25.31, 23.57,22.93, 22.31, 17.78, 17.08, 16.89, 15.55, 13.03. HRMS (m / z) [M + H] + calcd forC 45 H 75 NO 12 : 822.5362; found: 822.5344.

[0116] Example 20 (Preparation of compound HSC-20)

[0117] The preparation method of compound HSC-20 is the same as that of compound HSC-1, except that 3-aminomethylpyridine (28.8 mg, 0.26 mmol) is replaced with isopentylamine (23 mg, 0.26 mmol), and 73 mg of white solid powder is obtained, with a yield of 67%.

[0118] 1 H NMR (600 MHz, DMSO- d 6) δ 7.15 (t, J = 5.7 Hz, 1H), 5.20 (d, J = 3.8Hz, 1H, H12), 5.06 (d, J = 1.6 Hz, 1H, H1 of rha), 4.33 (d, J = 6.1 Hz, 1H, H1of ara), 3.34 – 3.27 (m, 2H), 1.58 – 1.54 (m, 2H), 1.09 (s, 3H), 1.07 (d, J =6.2 Hz, 3H), 0.88 (s, 3H), 0.87 (s, 6H), 0.85 (d, J = 1.5 Hz, 3H), 0.84 (d, J = 1.5 Hz, 3H), 0.67 (s, 3H), 0.57 (s, 3H).

[0119] 13 C NMR (151 MHz, DMSO) δ 176.04, 144.17, 121.41, 102.97, 99.94,79.30, 74.22, 72.85, 72.00, 70.41, 70.33, 68.14, 67.82, 64.41, 59.78, 47.13,46.19, 46.05, 45.19, 42.26, 41.32, 40.49, 40.06, 38.86, 38.20, 37.04, 36.92,35.97, 33.67, 32.96, 32.75, 32.00, 30.45, 26.96, 25.64, 25.31(×2), 23.60,22.48(×2),20.78, 17.78, 16.91, 15.56, 14.10, 13.02. HRMS(m / z)[M + H] + calcdfor C 46 H77 NO 11 : 820.5569; found: 820.5550.

[0120] Example 21 (Preparation of compound HSC-21)

[0121] Oleanolic acid (200 mg, 0.44 mmol), HoBt (118 mg, 0.88 mmol), and DIPEA (169 mg, 1.32 mmol) were dissolved in DMF (2 mL). After stirring at room temperature for 5 minutes, EDCI (168 mg, 0.88 mmol) was added and stirred for 30 minutes. Then, 3-methoxypropylamine (78 mg, 0.88 mmol) was added and the reaction was carried out overnight. The reaction was detected by TLC. After the reaction was completed, the reaction solution was poured into 15 mL of ice water, extracted twice with ethyl acetate (40 mL), the organic layer was washed twice with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure to obtain the crude product. Preparative liquid chromatography (methanol:water = 80:30) yielded 194 mg of white solid powder, with a yield of 84%.

[0122] 1 H NMR (400 MHz, DMSO- d 6) δ 7.27 (t, J = 5.6 Hz, 1H), 5.28 – 5.09 (m,1H), 3.29 (t, J = 6.3 Hz, 2H), 3.21 (s, 3H), 3.09 (m, 1H), 3.03 – 2.94 (m,2H), 2.76 (dd, J = 13.5, 4.4 Hz, 1H), 1.89 (td, J = 14.5, 3.7 Hz, 1H), 1.80(dd, J = 9.2, 3.7 Hz, 2H), 1.21 – 1.15 (m, 1H), 1.08 (s, 3H), 0.89 (s, 3H), 0.88 (s, 3H), 0.86 (s, 3H), 0.84 (s, 3H), 0.67 (s, 3H), 0.66 (s, 3H).HRMS (m / z) [M + H] + calcd for C 34 H 57 NO3: 528.4411; found: 528.4408.

[0123] Example 22 (Preparation of compound HSC-22)

[0124] The preparation method of compound HSC-22 is the same as that of compound HSC-21, except that 3-methoxypropylamine (78 mg, 0.88 mmol) is replaced with 9-aminofluorene (158 mg, 0.88 mmol), yielding 214 mg of white solid powder, with a yield of 79%. HRMS (m / z) [M + H] + calcd for C 43 H 57 NO2: 620.4462; found: 620.4471.

[0125] Example 23 (Preparation of compound HSC-23)

[0126] The preparation method of compound HSC-23 is the same as that of compound HSC-21, except that 3-methoxypropylamine (78 mg, 0.88 mmol) is replaced with 3-aminomethyl-tetrahydrofuran (88 mg, 0.88 mmol), yielding 153 mg of a white solid powder, with a yield of 65%. HRMS (m / z) [M + H] + calcd for C 35 H 57 NO3: 540.4411; found: 540.4419.

[0127] Example 24 (Preparation of compound HSC-24)

[0128] Oleanolic acid (200 mg, 0.44 mmol) was dissolved in DMF (3 mL), potassium carbonate (181 mg, 1.32 mmol) was added, followed by 2-(bromomethyl)-3,5,6-trimethylpyrazine (141 mg, 0.66 mmol). The reaction was carried out at 50 °C for 12 hours. After the reaction was completed, the reaction solution was poured into 15 mL of ice water, extracted twice with ethyl acetate (40 mL), the organic layer was washed twice with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure to obtain the crude product. Purification by preparative liquid chromatography (methanol:water = 80:30) yielded 191 mg of a white solid powder, with a yield of 74%.

[0129] 1 HNMR (400 MHz, DMSO- d6) δ 5.13 (d, J = 12.3 Hz, 1H), 5.10 – 5.04 (m,2H), 2.97 (dd, J = 9.7, 5.8 Hz, 1H), 2.76 (dd, J = 13.9, 4.6 Hz, 1H), 2.46(s, 3H), 2.42 (s, 3H), 2.40 (s, 3H), 1.05 (s, 3H), 0.87 (s, 3H), 0.86 (s,6H), 0.79 (s, 3H), 0.66 (s, 3H), 0.35 (s, 3H).

[0130] 13 C NMR (151 MHz, DMSO) δ 176.08, 150.59, 148.65, 148.25, 145.00,143.22, 121.89, 76.72, 64.23, 54.73, 46.97, 46.17, 45.40, 41.16, 40.93,38.35, 38.03, 36.51, 33.17, 32.70, 32.27, 32.02, 30.35, 28.20, 26.97, 26.90,25.55, 23.32, 22.83, 22.65, 21.19, 20.95, 20.06, 17.91, 16.32, 16.03, 15.02,14.09.HRMS (m / z) [M + H] + calcd for C 38 H 58 N2O3: 591.4526; found: 591.4416.

[0131] Example 25 (Preparation of compound HSC-25)

[0132] The preparation method of compound HSC-25 is the same as that of compound HSC-21, except that 3-methoxypropylamine (78 mg, 0.88 mmol) is replaced with cyclopropylmethylamine (62 mg, 0.88 mmol), and 154 mg of white solid powder is obtained, with a yield of 69%.

[0133] 1 HNMR (400 MHz, DMSO- d 6) δ 7.17 (t, J = 5.7 Hz, 1H), 5.09 (d, J= 3.5Hz, 1H), 3.04 (s, 1H), 2.93 – 2.76 (m, 2H), 2.66 (ddd, J = 13.7, 6.6, 4.2 Hz,2H), 0.96 (s, 3H), 0.77 – 0.73 (m, 9H), 0.71 (s, 3H), 0.56 (s, 3H), 0.54 (s,3H), 0.20 (dt, J = 8.2, 2.9 Hz, 2H), 0.06 – 0.05 (m, 2H).

[0134] 13 C NMR (151 MHz, DMSO) δ 176.10, 144.14, 121.37, 76.76, 48.54, 47.12,46.07, 45.19, 45.18, 43.23, 43.10, 41.29, 40.50, 40.48, 38.87, 38.37, 38.08,36.58, 33.66, 32.94, 32.78, 32.47, 30.44, 28.24, 26.94, 25.64, 23.55, 22.95,22.30, 18.01, 16.89, 16.03, 15.10, 11.01.HRMS (m / z) [M + H] + calcd for C 34 H 55 NO2:510.4305; found: 510.4312.

[0135] Example 26 (Preparation of compound HSC-26)

[0136] The preparation method of compound HSC-26 is the same as that of compound HSC-21, except that 3-methoxypropylamine (78 mg, 0.88 mmol) is replaced with 2-cyclopropylethylamine hydrochloride (106 mg, 0.88 mmol), and 153 mg of white solid powder is obtained, with a yield of 67%.

[0137] 1 HNMR (400 MHz, Chloroform- d ) δ 6.06 (dd, J = 6.9, 4.0 Hz, 1H), 5.36(t, J = 3.6 Hz, 1H), 3.52 (dq, J= 13.5, 6.8 Hz, 1H), 3.21 (dd, J = 11.0, 4.7Hz, 1H), 3.02 (dtd, J = 13.6, 7.1, 4.0 Hz, 1H), 2.49 (dd, J = 13.2, 4.4 Hz, 1H), 1.95 (d, J = 3.9 Hz, 1H), 1.90 (dd, J = 9.1, 3.6 Hz, 2H), 1.59 (d, J =3.1 Hz, 1H), 1.15 (s, 3H), 0.98 (s, 3H), 0.90 (d, J = 1.4 Hz, 9H), 0.77 (s, 3H), 0.76 (s, 3H).

[0138] 13 C NMR (151 MHz, CDCl3) δ 177.97, 145.07, 122.63, 78.86, 55.02,47.47, 46.78, 46.15, 42.25, 42.00, 39.60, 39.28, 38.69, 38.39, 36.88, 34.07,34.06, 32.92, 32.34, 32.24, 30.66, 28.01, 27.21, 27.07, 25.70, 23.73, 23.47(×2), 18.21, 16.77, 15.48, 15.29, 8.68, 4.34, 3.99.HRMS (m / z) [M + H] + calcdfor C 35 H 57 NO2: 524.4462; found: 524.4462.

[0139] Example 27 (Preparation of compound HSC-27)

[0140] The preparation method of compound HSC-27 is the same as that of compound HSC-21, except that 3-methoxypropylamine (78 mg, 0.88 mmol) is replaced with ethylamine hydrochloride (71 mg, 0.88 mmol), and 165 mg of white solid powder is obtained, with a yield of 78%.

[0141] 1 HNMR (400 MHz, DMSO- d6) δ 7.25 (t, J = 5.6 Hz, 1H), 5.36 – 5.05 (m,1H), 3.07 (ddd, J = 12.9, 6.6, 1.5 Hz, 1H), 3.01 – 2.95 (m, 2H), 2.78 (dd, J = 13.5, 4.4 Hz, 1H), 1.08 (s, 3H), 0.95 (t, J = 7.1 Hz, 3H), 0.89 (s, 3H), 0.87 (s, 3H), 0.86 (s, 3H), 0.84 (s, 3H), 0.67 (d, J = 1.7 Hz, 6H).

[0142] 13 C NMR (151 MHz, DMSO) δ 175.92, 144.11, 121.36, 76.77, 54.81, 47.11,46.06, 45.05, 41.23, 40.39, 38.92, 38.37, 38.07, 36.58, 33.65, 33.56, 33.44,32.95, 32.75, 32.43, 30.45(×2), 28.24, 26.93, 25.68(×2), 23.56, 22.92,18.00, 16.85, 16.04, 15.09.HRMS (m / z) [M + H] + calcd for C 32 H 53 NO2: 484.4149; found: 484.4155.

[0143] Example 28 (Preparation of compound HSC-28)

[0144] The preparation method of compound HSC-28 is the same as that of compound HSC-21, except that 3-methoxypropylamine (78 mg, 0.88 mmol) is replaced with 2,2-difluoroethylamine (71 mg, 0.88 mmol), and 138 mg of white solid powder is obtained, with a yield of 61%.

[0145] 1 H NMR (600 MHz, DMSO- d 6) δ 7.72 (t, J = 5.8 Hz, 1H), 5.88 (tt, J=56.7, 4.3 Hz, 1H), 5.19 (t, J = 3.7 Hz, 1H), 3.52 – 3.39 (m, 2H), 2.80 (dd, J = 13.7, 4.6 Hz, 1H), 1.82 – 1.77 (m, 2H), 1.65 (t, J = 13.5 Hz, 1H), 1.31 (m,2H), 1.19 (dt, J = 12.8, 3.2 Hz, 1H), 1.08 (s, 3H), 0.89 (s, 3H), 0.87 (s,3H), 0.87 (s, 3H), 0.84 (s, 3H), 0.67 (s, 3H), 0.65 (s, 3H).

[0146] 13 C NMR (151 MHz, DMSO) δ 177.35, 143.78, 121.54, 114.37, 76.78,54.82, 48.55, 47.09, 45.89, 45.45, 41.23, 40.35, 38.37, 38.09, 36.58, 33.53,32.88, 32.63, 32.46, 30.39, 28.23(×2), 26.93, 25.63, 23.47(×2), 22.91,22.15, 17.99, 16.57, 16.03, 15.10.HRMS (m / z) [M + H] + calcd for C 32 H 51 F2NO2:520.3960; found: 520.3961.

[0147] Example 29 (Preparation of compound HSC-29)

[0148] The preparation method of compound HSC-29 is the same as that of compound HSC-24, except that 2-(bromomethyl)-3,5,6-trimethylpyrazine (141 mg, 0.66 mmol) is replaced with (bromomethyl)cyclopropane (118 mg, 0.88 mmol), and 167 mg of white solid powder is obtained, with a yield of 75%.

[0149] 1 HNMR (400 MHz, Chloroform- d ) δ 5.29 (t, J= 3.7 Hz, 1H), 3.85 (ddd, J = 49.9, 11.4, 7.1 Hz, 2H), 3.21 (dd, J = 11.3, 4.7 Hz, 1H), 2.87 (dd, J =14.0, 4.6 Hz, 1H), 1.13 (s, 3H), 0.98 (s, 3H), 0.92 (s, 3H), 0.89 (s, 6H), 0.77 (s, 3H), 0.73 (s, 3H), 0.55 – 0.48 (m, 2H), 0.25 (dt, J = 6.0, 4.6 Hz, 2H).

[0150] 13 C NMR (151 MHz, CDCl3) δ 177.94, 143.96, 122.46, 79.17, 68.84,55.36, 47.77, 46.82, 46.07, 41.88, 41.47, 39.46, 38.89, 38.59, 37.18, 34.07,33.27, 32.93, 32.62, 30.86, 28.25, 27.80, 27.34, 26.00, 23.78, 23.58, 23.16,18.49, 17.23, 15.72, 15.47, 9.97, 3.32, 3.22.HRMS (m / z) [M + H] + calcd forC 34 H 54 O3: 511.41457; found: 511.4159.

[0151] Example 30 (Preparation of compound HSC-30)

[0152] The preparation method of compound HSC-30 is the same as that of compound HSC-21, except that 3-methoxypropylamine (78 mg, 0.88 mmol) is replaced with 4-phenoxybenzylamine (174 mg, 0.88 mmol), yielding 220 mg of a white solid powder, with a yield of 79%. HRMS (m / z) [M + H] + calcd for C 43 H 59 NO3: 638.4567; found: 638.4571.

[0153] Example 31 (Preparation of compound HSC-31)

[0154] The preparation method of compound HSC-31 is the same as that of compound HSC-21, except that 3-methoxypropylamine (78 mg, 0.88 mmol) is replaced with 3-aminomethylpyridine (94 mg, 0.88 mmol), yielding 170 mg of a white solid powder, with a yield of 71%. HRMS (m / z) [M+ H] + calcd for C 36 H 54 N2O2: 547.4258; found: 547.4259.

[0155] II. Efficacy Test

[0156] 1. In vitro anti-inflammatory effects of the target compound

[0157] RAW264.7 cells were cultured at a rate of 4.5 × 10⁻⁶. 4 RAW 264.7 cells were seeded at a density of 400 μL / well in 48-well plates and cultured overnight in RAW 264.7 cell culture medium. Cells were then divided into a blank control group, a lipopolysaccharide (LPS) group, and three drug dose groups, with three replicates per group. The drugs used in the drug groups included senna saponin B5 (HSC) and a pentacyclic triterpenoid derivative synthesized in this invention. Each drug dose group received 400 μL of culture medium at final drug concentrations of 0 μM, 2.3 μM, 6.7 μM, 20 μM, and 60 μM, respectively, and were incubated for 2 h. After incubation, the original culture medium was removed. The blank group received fresh complete culture medium, while the other groups received 400 μL of LPS at a final concentration of 1 μg / mL. After 24 h of incubation, the cell supernatant was used for ELISA to detect the inhibitory effect of each derivative on the release of the inflammatory cytokine IL-6 induced by LPS from RAW 264.7 cells. The results are shown in Table 1 below.

[0158] Table 1. Levels of the inflammatory factor IL-6 in each group

[0159]

[0160] Table 1 shows that Pulsatilla saponin B5 (HSC) and the pentacyclic triterpenoid derivative synthesized in this invention can significantly inhibit the level of IL-6, an inflammatory factor induced by LPS in RAW264.7 cells. Among them, compared with HSC, compounds HSC-4, HSC-9, HSC-14, and HSC-20 all showed superior anti-inflammatory activity, with HSC-4 and HSC-14 being the most significant.

[0161] 2. Cytotoxic effects of the target compound

[0162] RAW264.7 cells were cultured at a concentration of 1.5 × 10⁻⁶. 4 Cells were seeded at a density of 100 μL / well in 96-well plates and cultured overnight in RAW 264.7 cell culture medium. Cells were then divided into a blank control group and drug groups at various dosages, with three replicates per group. The drug used in the drug groups was a pentacyclic triterpenoid derivative synthesized in this invention. 100 μL of culture medium was added to each well, with final drug concentrations of 0 μM, 3.9 μM, 7.8 μM, 15.6 μM, 31.3 μM, 62.5 μM, and 125 μM. The wells were incubated for 24 h. After incubation, 10 μL of CCK-8 reagent was added to each well, and the cells were incubated at 37°C in the dark for 4 h. The absorbance of each well was measured at 450 nm using a microplate reader, and the cell viability was calculated. The cytotoxicity of each derivative was also assessed. The results are shown in Table 2 below.

[0163] Table 2 Cell viability of each group

[0164]

[0165] Table 2 shows that the pentacyclic triterpenoid derivatives synthesized in this invention have no obvious toxicity at concentrations below 62.5 μM (cell survival rate >85%).

[0166] 3. Therapeutic effects of the dominant compound on LPS-induced sepsis in mice

[0167] SPF-grade, 6-8 week old, healthy male C57BL / 6 mice were acclimatized for three days and then randomly divided into 8 groups (n=9) according to body weight: control group, model group, positive control group (dexamethasone, 5 mg / kg), scutellaria baicalensis saponin B5 group (HSC, 20 mg / kg), low-dose BSC-4 group (10 mg / kg), high-dose BSC-4 group (20 mg / kg), low-dose BSC-14 group (10 mg / kg), and high-dose BSC-14 group (20 mg / kg). Scutellaria baicalensis saponin B5 and its derivative dosage groups were administered intraperitoneally 1 hour before and 1 hour after modeling, with a dosage of 10 mg / mL, for a total of two administrations. The dexamethasone group was administered intraperitoneally 1 hour after modeling, with a dosage of 10 mg / mL, for a total of one administration. Except for the blank control group, all other groups were intraperitoneally injected with 4 mg / kg LPS for modeling. Mice were sacrificed 12 hours after modeling, and blood, heart, lungs, and intestines were collected. Serum and tissue inflammatory factor IL-6 levels were detected using an ELISA kit.

[0168] Twelve hours after modeling, except for the normal control group mice which showed no significant changes, the other groups all exhibited symptoms such as poor mental state, rough fur, decreased body temperature, trembling, closed eyes, increased eye discharge and anal excretion. The condition of the mice in the drug-treated group was better than that of the model group.

[0169] Histopathological analysis results of each group are as follows: Figure 2 As shown in the HE staining (hematoxylin-eosin staining) results, after LPS intraperitoneal injection for modeling, the cardiac tissue of the model group showed loose cytoplasm of cardiomyocytes in the ventricular wall, with many small round vacuoles in the cytoplasm of cardiomyocytes, and a large number of cardiomyocyte nuclei with irregular shapes and dark cytoplasm staining. Figure 2 A); The alveolar walls of the lung tissue show extensive granulocyte infiltration, significant thickening of the alveolar walls, widening of the alveolar septa, narrowing of many alveoli, and varying alveolar sizes; the bronchiolar mucosal epithelial cells are irregularly arranged, and a large amount of eosinophilic material and desquamated epithelial cells are visible in the lumen. Figure 2 B); In the colon tissue, dilated intestinal glands with enlarged gland lumens, flattened epithelial cells, necrosis of intestinal gland epithelial cells with fragmented nuclei, abundant goblet cells, and a large number of free necrotic cell fragments can be seen in the intestinal lumen. Figure 2 C); In the positive drug dexamethasone group, HSC group, and HSC-4 and HSC-14 dosage groups, the inflammatory symptoms of the heart, lungs, and intestines were significantly relieved.

[0170] The levels of the inflammatory factor IL-6 in the heart, lung, and intestinal tissues of each group were detected, and the results are shown in Table 3. LPS modeling significantly increased the IL-6 levels in the heart, lung, and intestinal tissues (P<0.05, P<0.005). The positive control drugs dexamethasone, HSC, and HSC-4 and HSC-14 at all doses significantly reduced the IL-6 levels in the heart, lung, and intestinal tissues, and the anti-inflammatory effects of HSC-4 and HSC-14 were similar to those of dexamethasone.

[0171] Table 3. Levels of IL-6, an inflammatory factor, in the heart, lungs, and intestines of each group.

[0172]

[0173] Finally, it should be noted that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A pentacyclic triterpenoid derivative, characterized in that: The pentacyclic triterpenoid derivatives are selected from the following structural formulas: 。 2. The use of a pentacyclic triterpenoid derivative according to claim 1 in the preparation of an anti-inflammatory drug.

3. A pharmaceutical composition, characterized in that: Includes the pentacyclic triterpenoid derivative of claim 1 or a pharmaceutically acceptable salt thereof.

4. A pharmaceutical preparation, characterized in that: It includes a therapeutically effective amount of the pentacyclic triterpenoid derivative of claim 1 or a pharmaceutically acceptable salt thereof, as well as a pharmaceutically acceptable carrier or excipient.

5. A method for preparing the pentacyclic triterpenoid derivative according to claim 1, characterized in that: Includes the following steps: The pentacyclic triterpenoid derivatives were prepared by esterification or amidation reactions using α-hederone as the starting material.

6. The preparation method according to claim 5, characterized in that: The esterification reaction includes the following steps: using α-hederine as the starting material, in the presence of potassium carbonate and solvent, an esterification reaction is carried out with a halogenated compound to generate a pentacyclic triterpene derivative; The molar ratio of the α-hederone to the halogenated compound is 1:(1-2).

7. The preparation method according to claim 5, characterized in that: The amidation reaction includes the following steps: using α-hederine as the starting material, in the presence of auxiliary reagents and solvents, an amidation reaction is carried out with amine compounds to generate pentacyclic triterpenoid derivatives; The molar ratio of α-hederone to amine compounds is 1:(2-3).

8. The preparation method according to claim 7, characterized in that: The auxiliary reagent is any one or more of 1-hydroxybenzotriazole, N,N-diisopropylethylamine, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, N,N'-carbonyldiimide, dicyclohexylcarbodiimide, diisopropylcarbodiimide, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, O-benzotriazole-tetramethylurea hexafluorophosphate, 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate, 2-succinimide-1,1,3,3-tetramethylurea tetrafluoroborate, N'-tetramethylurea tetrafluoroborate, diphenyl azidophosphate, 4-dimethylaminopyridine, and 1-hydroxy-7-azabenzotriazole.

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