18beta-glycyrrhetinic acid derivative, and preparation method and application thereof

Abstract

The application discloses an 18beta-glycyrrhetinic acid derivative, a preparation method and application thereof, and belongs to the technical field of medicines, wherein a structural formula of the 18beta-GA derivative is shown in the specification. The E ring 30-COOH and the A ring 3-OH of the 18beta-GA are modified by taking the 18beta-GA as a leading compound. The modification of the C3-OH group of the 18beta-GA can change the molecular polarity of the 18beta-GA, so that the better cytotoxicity and anti-proliferation activity advantages of the compound are realized. The amidation of the C-30 carboxyl group can enhance the anti-tumor efficacy of the 18beta-GA derivative. The application introduces a polyamine compound at the 30th position, and then introduces a piperazine polyamine compound at the 30th position, and then the 3rd hydroxyl group is esterified and connected with different small molecule Boc-amino acid esters. The application obtains a compound with high efficient anti-triple negative breast cancer effect and good drug property by scientific structural modification of the 18beta-GA.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to an 18β-glycyrrhetinic acid derivative, its preparation method, and its application. Background Technology

[0002] Triple-negative breast cancer (TNBC) refers to breast cancer in which immunohistochemical examination of the cancer tissue shows negative results for estrogen receptor, progesterone receptor, and the proto-oncogene Her-2. This type of breast cancer accounts for 10.0% to 20.8% of all breast cancer pathological types. It has unique biological behavior and clinicopathological features, and its prognosis is worse than other types. As a prominent subtype of breast cancer, triple-negative breast cancer will undoubtedly become a major clinical challenge in the field of cancer medicine.

[0003] Glycyrrhizic acid (GL) is one of the most important components of licorice. It is a β-amyrin-type triterpenoid saponin composed of two molecules of glucuronic acid and one molecule of glycyrrhetinic acid (GA). The pharmacological properties of licorice are partly attributed to GL. However, GL is not directly absorbed through the gastrointestinal tract. Instead, it is hydrolyzed by intestinal bacteria into 18β-glycyrrhetinic acid (18β-GA). After intestinal absorption, 18β-GA is metabolized in the liver to 3β-monoglucuronic acid-18β-GA, and this metabolite circulates in the blood. Therefore, the oral bioavailability of GL and 18β-GA is relatively poor. In nature, two forms of GA exist, mainly based on the different spatial conformations of H-18: 18α-GA and 18β-GA. The content of 18β-GA in plants is much higher than that of 18α-GA. Therefore, modern research mainly focuses on 18β-GA.

[0004] Recent studies have shown that 18β-GA possesses a variety of significant biological activities, including anti-inflammatory, antiviral, hepatoprotective, and antitumor effects. 18β-GA exhibits significant cytotoxic activity in vitro against various cell types, including breast cancer cells, human hepatocellular carcinoma cells, non-small cell lung cancer cells, prostate cancer cells, and colorectal cancer cells. Studies have also demonstrated good anti-breast cancer efficacy for 18β-GA and its derivatives, such as the anti-breast cancer MCF-7 derivative. 18β-GA has also shown noteworthy activity in various in vivo experimental cancer models, and it is known to trigger apoptosis in tumor cell lines. The significant antitumor activity of 18β-GA has remained a research hotspot both domestically and internationally.

[0005] In recent years, researchers at home and abroad have made great progress in addressing many issues such as the structural modification, pharmacological activity, and structure-activity relationship of 18β-GA. Its structural modification can be divided into four categories: α-ring, β-ring, β-ring and polycyclic modification. (1) Introducing hydroxyl groups such as aminoalkyl, amino acid, and sugar groups into C-3 through esterification; (2) Oxidation or elimination of hydroxyl groups at C-3, introduction of functional groups at C-2, opening or increasing the number of atoms in α-ring; (3) Elimination of ketone group at C-11 on β-ring; (4) Esterification or amidation of carboxyl group at C-30 on β-ring; (5) Simultaneous esterification of hydroxyl group at C-3 and carboxyl group at C-30, disappearance of ketone group at C-11, and simultaneous esterification at C-30. However, the structural modification research of 18β-GA has the following problems: the synthetic route is too long, the synthetic steps are relatively complicated, the reaction conditions are harsh, and it may even completely destroy the pentacyclic triterpenoid structure of GA.

[0006] Researchers such as George Pak-Heng Leung discovered that 18β-GA, in combination with doxorubicin, exhibited significant cytotoxicity, apoptosis, and loss of mitochondrial membrane potential in MCF-7 breast cancer cells, achieved through the upregulation of the mitochondrial-dependent apoptosis pathway. However, because 18β-GA inhibits type 2 11β-hydroxysteroid dehydrogenase, long-term high-dose use of 18β-GA often causes pseudoaldosteronism, clinically manifested as hypertension, hypokalemia, and other adverse reactions. Research utilizing 18β-GA as a scaffold to develop novel, low-toxicity, and highly effective antitumor drugs has attracted increasing attention. Gaber O. Moustafa et al. combined 18β-GA with various amino acids to synthesize eight novel 18β-GA-amino acid peptides and peptide ester derivatives, and conducted in vitro cytotoxicity activity tests on the new compounds, which did not show excellent inhibitory effects on breast cancer. Hongke Liu et al. directly amide-linked a nitrogen-containing aromatic compound to the -COOH group at the 30 position of 18β-GA, obtaining a novel arylruthenium complex of 18β-GA derivative ligands with Ru(II). Their antitumor activity was evaluated and showed only moderate inhibitory activity. Li et al. modified the structure at the 3 and 30 positions of 18β-GA, obtaining the compound with the best anti-triple-negative breast cancer activity to date, with an IC50 value of [missing information]. 50 The value was 1.37 ± 0.18 μM.

[0007] Breast cancer is one of the most common and deadliest cancers among women worldwide. Currently, all inventions involving the modification of the 18β-GA structure have only shown inhibitory activity against common breast cancer, and no inventions have focused on the development of inhibitors for refractory triple-negative breast cancer.

[0008] Therefore, how to develop an 18β-GA derivative, its preparation method, and its application in the preparation of drugs for treating triple-negative breast cancer are problems that urgently need to be solved by those skilled in the art. Summary of the Invention

[0009] In view of this, the present invention provides an 18β-GA derivative, its preparation method and application.

[0010] To achieve the above objectives, the present invention adopts the following technical solution:

[0011] An 18β-GA derivative having the structure shown in Formula 1-5 or Formula I-II:

[0012]

[0013]

[0014] In Formula I, R1 is selected from 3-phenylpropionyl, Fmoc-L-methionine acyl, Boc-glycine acyl, Boc-L-leucine acyl, Boc-L-methionine acyl, Boc-L-isoleucine acyl, Boc-L-phenylalanine acyl, Boc-L-proline acyl, Boc-N-methyl-L-alanine acyl, Boc-L-valine acyl, 4-benzyl ester-Boc-L-aspartic acyl, Boc-L-tryptophan acyl, N-(tert-butoxycarbonyl)-N-methylglycine acyl or Boc-L-pyroglutamic acid acyl;

[0015] In Formula II, R2 is selected from Boc-glycine acyl, Boc-L-leucine acyl, Boc-L-methionine acyl, Boc-L-isoleucine acyl, Boc-L-phenylalanine acyl, Boc-L-proline acyl, Boc-N-methyl-L-alanine acyl, Boc-L-valine acyl, 4-benzyl ester-Boc-L-aspartic acid acyl, Boc-L-tryptophan acyl, N-(tert-butoxycarbonyl)-N-methylglycine acyl or Boc-L-pyroglutamic acid acyl.

[0016] The present invention also provides a method for preparing the above-mentioned 18β-GA derivative, wherein the method for preparing the compound shown in Formula 4 includes the following steps:

[0017] (1) Weigh 18β-GA, EDCI and HOBT and place them in a reaction vessel. Add anhydrous dichloromethane to dissolve them, then add TEA dropwise. Stir at room temperature to carry out the reaction. Monitor the reaction system by thin-layer chromatography. Stop stirring after the reaction is complete to obtain the reactants.

[0018] (2) Pour the reactants into ice water, transfer the mixture to a separatory funnel, extract with dichloromethane, collect the organic phase, extract with saturated sodium chloride aqueous solution, collect the organic phase, dry with anhydrous sodium sulfate, filter, concentrate under reduced pressure to obtain the target crude product, separate and purify the target crude product by silica gel column chromatography, concentrate under reduced pressure to obtain the compound shown in Formula 4.

[0019] Furthermore, in step (1), the molar ratio of 18β-GA, EDCI, HOBT and TEA is 0.43∶0.86∶0.43∶2.15.

[0020] Furthermore, the preparation method of the compounds shown in Formulas 1-3 includes the following steps:

[0021] (1.1) Weigh 18β-GA, EDCI and HOBT and place them in a reaction vessel. Dissolve them in anhydrous dichloromethane, then add TEA dropwise. Stir at room temperature, add the polyamine compound, and stir at room temperature to carry out the reaction. Monitor the reaction system by thin-layer chromatography. Stop stirring after the reaction is complete to obtain the reactants. The polyamine compound is piperazine-1-carboxylic acid methyl ester, N-methylpiperazine or N,N-dimethyldipropyltriamine.

[0022] (1.2) Pour the reactants into ice water, transfer the mixture to a separatory funnel, extract with dichloromethane, collect the organic phase, extract with saturated sodium chloride aqueous solution, collect the organic phase, dry with anhydrous sodium sulfate, filter, concentrate under reduced pressure to obtain the target crude product, separate and purify the target crude product by silica gel column chromatography, concentrate under reduced pressure to obtain the compounds shown in Formulas 1-3.

[0023] Furthermore, in step (1.1), the molar ratio of 18β-GA, EDCI, HOBT, TEA and the polyamine compound is 0.43∶0.86∶0.43∶2.15∶2.15.

[0024] Furthermore, the preparation method of the compound shown in Formula 5 includes the following steps:

[0025] (2.1) Weigh L-methionine and place it in a reaction vessel. Add acetonitrile to dissolve it. Then weigh the compound shown in Formula 4 and dissolve it in anhydrous dichloromethane. Add the dichloromethane solution of the compound shown in Formula 4 to the acetonitrile solution of L-methionine. Add TEA dropwise and stir at room temperature to carry out the reaction. Monitor the reaction system by thin-layer chromatography. Stop stirring after the reaction is completed to obtain the reactants.

[0026] (2.2) Pour the reactants into ice water, transfer the mixture to a separatory funnel, extract with dichloromethane, collect the organic phase, extract with saturated sodium chloride aqueous solution, collect the organic phase, dry with anhydrous sodium sulfate, filter, concentrate under reduced pressure to obtain the target crude product, purify the target crude product by recrystallization, concentrate under reduced pressure to obtain the compound shown in Formula 5.

[0027] Furthermore, in step (2.1), the molar ratio of L-methionine, the compound shown in Formula 4, and triethylamine is 2.00:0.40:4.00.

[0028] Furthermore, the preparation method of the compound shown in Formula I includes the following steps:

[0029] (3.1) Weigh the compound shown in Formula 1, EDCI and DMAP and place them in a reaction vessel. Add anhydrous dichloromethane to dissolve them. Then weigh 3-phenylpropionic acid, Fmoc-L-methionine, Boc-glycine, Boc-L-leucine, Boc-L-methionine, Boc-L-isoleucine, Boc-L-phenylalanine, Boc-L-proline, Boc-N-methyl-L-alanine, Boc-L-valine, Boc-L-aspartic acid-4-benzyl ester, Boc-L-tryptophan, N-(tert-butoxycarbonyl)-N-methylglycine or Boc-L-pyroglutamic acid and add them to the above mixture to dissolve them. Continue to stir the mixture at room temperature to carry out the reaction. Monitor the reaction system by thin-layer chromatography. Stop stirring after the reaction is completed to obtain the reactants.

[0030] (3.2) Pour the reactants into ice water, transfer the mixture to a separatory funnel, extract with dichloromethane, collect the organic phase, extract with saturated sodium chloride aqueous solution, collect the organic phase, dry with anhydrous sodium sulfate, filter, concentrate under reduced pressure to obtain the target crude product, separate and purify the target crude product by silica gel column chromatography, concentrate under reduced pressure to obtain the compound shown in Formula I.

[0031] In step (3.1), the molar ratio of the compound shown in Formula 1, EDCI, DMAP, 3-phenylpropionic acid, Fmoc-L-methionine, Boc-glycine, Boc-L-leucine, Boc-L-methionine, Boc-L-isoleucine, Boc-L-phenylalanine, Boc-L-proline, Boc-N-methyl-L-alanine, Boc-L-valine, Boc-L-aspartic acid-4-benzyl ester, Boc-L-tryptophan, N-(tert-butoxycarbonyl)-N-methylglycine, or Boc-L-pyroglutamic acid is 1.00∶1.20∶0.20∶1.20.

[0032] Furthermore, the preparation method of the compound shown in Formula II includes the following steps:

[0033] (4.1) Weigh the compound shown in Formula 2, EDCI and DMAP and place them in a reaction vessel. Add anhydrous dichloromethane to dissolve them. Then weigh Boc-glycine, Boc-L-leucine, Boc-L-methionine, Boc-L-isoleucine, Boc-L-phenylalanine, Boc-L-proline, Boc-N-methyl-L-alanine, Boc-L-valine, Boc-L-aspartic acid-4-benzyl ester, Boc-L-tryptophan, N-(tert-butoxycarbonyl)-N-methylglycine or Boc-L-pyroglutamic acid and add them to the above mixture to dissolve them. Continue to stir the mixture at room temperature to carry out the reaction. Monitor the reaction system by thin-layer chromatography. Stop stirring after the reaction is completed to obtain the reactants.

[0034] (4.2) Pour the reactants into ice water, transfer the mixture to a separatory funnel, extract with dichloromethane, collect the organic phase, extract with saturated sodium chloride aqueous solution, collect the organic phase, dry with anhydrous sodium sulfate, filter, concentrate under reduced pressure to obtain the target crude product, separate and purify the target crude product by silica gel column chromatography, concentrate under reduced pressure to obtain the compound shown in Formula I.

[0035] In step (4.1), the molar ratio of the compound shown in Formula 2, EDCI, DMAP, and Boc-glycine, Boc-L-leucine, Boc-L-methionine, Boc-L-isoleucine, Boc-L-phenylalanine, Boc-L-proline, Boc-N-methyl-L-alanine, Boc-L-valine, Boc-L-aspartic acid-4-benzyl ester, Boc-L-tryptophan, N-(tert-butoxycarbonyl)-N-methylglycine, or Boc-L-pyroglutamic acid is 1.00∶1.20∶0.20∶1.20.

[0036] The present invention also provides the application of the above-mentioned 18β-GA derivative in the preparation of a drug for treating triple-negative breast cancer.

[0037] The beneficial effects of this invention are:

[0038] This invention develops compounds with highly efficient anti-triple-negative breast cancer activity using a green and mild synthetic method. 18β-GA is used as a lead compound, and its E ring 30-COOH and A ring 3-OH are structurally modified. Modification of the C3-OH group of 18β-GA alters its molecular polarity, resulting in better cytotoxicity and anti-proliferative activity. The usual amidation of the C-30 carboxyl group enhances the antitumor efficacy of the 18β-GA derivatives. This invention introduces polyamine compounds (piperazines, methionine, and N,N-dimethyldipropyltriamine) at the 30-position carboxyl group to form the GA-NH series of derivatives. Then, based on the introduction of piperazine and other polyamine compounds at the 30-position, the hydroxyl group at the 3-position is esterified with different small-molecule Boc-amino acids to form the GA-NH-OO series of derivatives (with an ester bond at the C-3 position). This invention tested the antiproliferative activity of the obtained series of derivatives, the positive control drug doxorubicin, and the lead compound 18β-GA against the human triple-negative breast cancer cell line MDA-MB-231 using the MTT assay. Most of the obtained derivatives showed stronger activity than the parent compound; the compound shown in Formula 14 exhibited stronger antiproliferative activity against triple-negative breast cancer cells than the positive control drug doxorubicin, and was 122.3 times stronger than the parent compound 18β-GA, with an IC50 value of [missing value]. 50 The value was 0.37±0.01 μM. This invention obtained candidate compounds with high efficacy against triple-negative breast cancer and good drug-like properties through scientific structural modification of 18β-GA. Among them, the compound represented by the dominant formula 14 exhibits stronger anti-triple-negative breast cancer cell proliferation activity than various 18β-GA derivatives reported to date.

[0039] This invention avoids the formation of byproducts from the reaction of 18β-GA with condensing agents or its own dimerization, which are difficult to remove during post-processing. During synthesis, the water-soluble condensing agent 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) and the highly efficient catalyst 4-dimethylaminopyridine (DMAP) facilitate the esterification reaction. The lone pair of electrons carried by the nitrogen atom on the dimethylamino group in the highly efficient catalyst 4-dimethylaminopyridine can resonate with the electron cloud of the pyridine ring, enhancing the nucleophilicity of the nitrogen atom on the pyridine ring. As an acylation transfer agent, 4-dimethylaminopyridine is much faster than pyridine-catalyzed acylation reactions. Furthermore, this invention eliminates the need to convert the carboxylic acid to an acyl halide before the reaction, thus improving the reaction yield.

[0040] This patent allows for the synthesis of 18β-GA derivatives in high yield through simple operations and mild conditions. Detailed Implementation

[0041] 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.

[0042] Example 1

[0043]

[0044] Weigh 0.43 mmol of 18β-GA, 0.86 mmol of EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), and 0.43 mmol of HOBT (1-hydroxybenzotriazole-hydrate) and place them in a 25 mL three-necked reactor. Dissolve them in 10 mL of anhydrous dichloromethane, then add 2.15 mmol of TEA (triethylamine) dropwise. Stir at room temperature for 2 h to activate the reaction. Add 2.15 mmol of piperazine-1-carboxylic acid methyl ester and stir at room temperature to carry out the reaction. Monitor the reaction system by thin-layer chromatography at regular intervals. Stop stirring when a large amount of the target product is generated and 18β-GA no longer decreases. Then pour the reaction mixture into a container. The reaction was terminated by adding 15 mL of ice water. The mixture was transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane. The organic phase was collected and then extracted twice with saturated sodium chloride aqueous solution to remove water-soluble impurities. The organic phase was collected and dried with anhydrous sodium sulfate to remove excess water. After filtration and concentration under reduced pressure, the target crude product was obtained. The target crude product was separated and purified by silica gel column chromatography (elution with petroleum ether:ethyl acetate = 10:1 to 1:1). After purification, the product was concentrated under reduced pressure to obtain the compound shown in Formula 1. The physical properties and characterization data of the obtained compound are as follows: yield: 50%; melting point: 151.2-153.1℃; white powder solid. 1HNMR(400MHz,CDCl3)δ5.66(s,1H,CH-12),3.71(s,3H,COOCH3),3.65-3.57(m,4H,N(CH2)2),3.46(t,J=5.3Hz,4H,N(CH2)2),3.21(dd,J=10.8,5.4Hz,1H,CH-3),2.77(dt,J=13.4,3.6Hz,1H,CH-1),2.32(s,1H,CH-9),2.31-2.25,2.07-2.01,1.96,1.83,1.71-1.55,1.41,1.40-1.37,0.88-0.83(18H,methylene and methine oftriterpenoidstructure),1.35(s,3H,CH3-29),1.21(s,3H,CH3-27),1.12(s,3H,CH3-25),1.11(s,3H,CH3-26),0.99(s,3H,CH3-23),0.80(s,3H,CH3-24),0.79(s,3H,CH3-28),0.68(dd,J=11.8,1.9Hz,1H,CH-5); 13 CNMR(101MHz,CDCl3)δ200.09(C-11),174.22(C-30),169.35(C-13),155.81(COOCH3),128.62(C-12),78.76(C-3),61.81(C-9),54.97(C-5),52.84(COOCH3),48.10(C-18),45.29(2C,N(CH2)2),43.93(2C,N(CH2)2),43.89(C-14),43.30(C-8),39.17(C-19),39.14(C-1),37.73(C-4),37.11(C-20 / C-22,2C),33.15(C-10),32.84(C-7),31.79(C-17),28.42(C-21),28.11(C-29),27.31(C-28),27.08(C-23),26.91(C-16),26.74(C-2),26.43(C-15),23.17(C-27),18.70(C-26),17.50(C-6),16.39(C-25),15.60(C-24);IRνmax 3487,2864,2789,1656,1629,1467,1388,1327,1284,1251,1188,1176,1124,1074,1041,767cm -1HRMS(ESI)m / z calcd for C 36 H 57 N2O5 + (M+H) + 597.4267, found 597.4283.

[0045] Example 2

[0046]

[0047] Weigh 0.43 mmol of 18β-GA, 0.86 mmol of EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), and 0.43 mmol of HOBT (1-hydroxybenzotriazole-hydrate) and place them in a 25 mL three-necked reactor. Dissolve them in 10 mL of anhydrous dichloromethane, then add 2.15 mmol of triethylamine dropwise. Stir at room temperature for 2 h to activate the reaction. Add 2.15 mmol of N-methylpiperazine and stir at room temperature to carry out the reaction. Monitor the reaction system by thin-layer chromatography at regular intervals. Stop stirring when a large amount of the target product is generated and 18β-GA no longer decreases. Then pour the reaction system into a 15 mL three-necked reactor. The reaction was terminated by soaking the mixture in 15 mL of ice water. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane. The organic phase was collected and then extracted twice with saturated sodium chloride aqueous solution to remove water-soluble impurities. The organic phase was collected and dried with anhydrous sodium sulfate to remove excess water. After filtration and concentration under reduced pressure, the target crude product was obtained. The target crude product was separated and purified by silica gel column chromatography (elution with dichloromethane:methanol = 20:1 to 12:1). After purification, the product was concentrated under reduced pressure to obtain the compound shown in Formula 2. The physical properties and characterization data of the obtained compound are as follows: yield: 88%; melting point: 249.2-252.8℃; white powder solid. 1H NMR(400MHz,CDCl3)δ5.71(s,1H,CH-12),3.64(t,J=4.9Hz,4H,N(CH2)2),3.22(dd,J=11.0,5.4Hz,1H,CH-3),2.79(dt,J=13.5,3.6Hz,1H,CH-1),2.39(t,J=5.0Hz,4H,N(CH2)2),2.33(s,1H,CH-9),2.30(s,3H,N-CH3),2.11-2.01,1.97,1.83,1.73-1.55,1.53-1.45,1.42,1.41-1.37,1.25,0.98-0.92,0.89-0.84(18H,methylene and methine oftriterpenoid structure),1.35(s,3H,CH3-29),1.21(s,3H,CH3-27),1.13(s,3H,CH3-25),1.12(s,3H,CH3-26),1.00(s,3H,CH3-23),0.81(s,3H,CH3-24),0.80(s,3H,CH3-28),0.69(dd,J=11.7,1.9Hz,1H,CH-5); 13 C NMR(101MHz,CDCl3)δ200.14(C-11),173.87(C-30),169.57(C-13),128.66(C-12),78.76(C-3),61.82(C-9),55.22(2C,N(CH2)2),54.98(C-5),48.17(C-18),45.99(N-CH3),45.30(2C,N(CH2)2),43.89(C-8),43.81(C-14),43.30(C-19),39.22(C-20),39.15(C-1),37.78(C-4),37.12(C-22),33.32(C-10),32.84(C-7),31.78(C-17),28.45(C-21),28.12(C-29),27.33(C-28),27.06(C-23),26.77(C-2),26.48(C-15 / C-16,2C),23.17(C-27),18.71(C-26),17.51(C-6),16.39(C-25),15.59(C-24);IRνmax2941,2864,2789,1658,1645,1460,1292,1140,1087,1074,781cm -1 .

[0048] Example 3

[0049]

[0050] Weigh 0.43 mmol of 18β-GA, 0.86 mmol of EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), and 0.43 mmol of HOBT (1-hydroxybenzotriazole-hydrate) and place them in a 25 mL three-necked reactor. Dissolve them in 10 mL of anhydrous dichloromethane, then add 2.15 mmol of triethylamine dropwise. Stir at room temperature for 2 h to activate the reaction. Add 2.15 mmol of N,N-dimethyldipropyltriamine and stir at room temperature to carry out the reaction. Monitor the reaction system by thin-layer chromatography at regular intervals. Stop stirring when a large amount of the target product is generated and 18β-GA no longer decreases. Then, proceed with the reaction... The system was poured into 15 mL of ice water to terminate the reaction. The mixture was transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane. The organic phase was collected and then extracted twice with saturated sodium chloride aqueous solution to remove water-soluble impurities. The organic phase was collected and then dried with anhydrous sodium sulfate to remove excess water. After filtration and concentration under reduced pressure, the target crude product was obtained. The target crude product was separated and purified by silica gel column chromatography (methanol:ethyl acetate = 3:1 elution). After purification, the product was concentrated under reduced pressure to obtain the compound shown in Formula 3. The physical properties and characterization data of the obtained compound are as follows: yield 41%; melting point: 106.5-108.8℃; white powder solid. 1H NMR(400MHz,CDCl3)δ7.30(t,J=4.7Hz,1H,CONH),5.65(s,1H,CH-12),3.40-3.28(m,2H,C’H-1),3.20(dd,J=10.9,5.1Hz,1H,CH-3),2.81-2.75(m,4H,C’H-3,C’H-4),2.40(t,J=6.6Hz,2H,C’H-6),2.31(s,1H,CH-9),2.24(s,6H,N(CH3)2),2.15,2.02,1.95,1.84,1.77-1.70,1.65-1.56,1.41,1.17,0.87-0.8(23H,methylene and methine oftriterpenoid structure and Methylene group of polyamine structure),1.36(s,3H,CH3-29),1.24(s,1H,NH),1.11(s,9H,CH3-25,CH3-26,CH3-27),0.99(s,3H,CH3-23),0.79(s,6H,CH3-24,CH3-28),0.68(d,J=11.3Hz,1H,CH-5); 13<13>C NMR (101 MHz, CDCl3) δ 200.02 (C-11), 175.93 (C-30), 169.43 (C-13), 128.45 (C-12), 78.71 (C-3), 61.84 (C-9), 58.14 (C'-6), 54.99 (C-5), 48.64 (C-18), 48.29 (C'-4), 48.14 (C'-3), 45.43 (2C, N(CH3)2), 45.38 (C-8), 43.48 (C-14), 43.22 (C-19), 41.80 (C-4), 39.26 (C'-1), 39.16 (C-20), 38.94 (C-1), 37.57 (C-10), 37.09 (C-22), 32.78 (C-7), 31.88 (C-17), 31.39 (C'-2), 29.71 (C'-5), 28.53 (C16), 28.30 (C-21), 28.14 (C-29), 27.30 (C-28), 27.06 (C-23), 26.51 (C-2), 26.47 (C-15), 23.41 (C-27), 18.68 (C-26), 17.50 (C-6), 16.36 (C-25), 15.62 (C-24); IR νmax 2943, 2929, 1654, 1535, 1465, 1386, 1280, 1255, 1041, 991, 792 cm -1 ; HRMS (ESI) m / z calcd for C 38 H 66 N3O3 + (M+H) + 612.50987, found 612.50989.

[0051] Example 4

[0052]

[0053] Weigh 0.43 mmol of 18β-GA, 0.86 mmol of EDCI, and 0.43 mmol of HOBT into a 25 mL three-necked reaction flask, dissolve them in 10 mL of anhydrous dichloromethane, and then add 2.15 mmol of triethylamine dropwise. Stir the mixture at room temperature, monitoring the reaction system periodically using thin-layer chromatography. Stop stirring when a large amount of the target product is formed and 18β-GA no longer decreases. Then pour the reaction mixture into 15 mL of ice water to terminate the reaction. Transfer the mixture to a separatory funnel, extract twice with 15 mL of dichloromethane, collect the organic phase, and then... The product was extracted twice with sodium chloride aqueous solution to thoroughly remove water-soluble impurities. The organic phase was collected, dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The target crude product was then separated and purified by silica gel column chromatography (elution with dichloromethane:ethyl acetate = 10:1 to 5:1). After purification, the product was concentrated under reduced pressure to obtain the compound shown in Formula 4. The physical properties and characterization data of the obtained compound are as follows: yield: 73%; melting point: 250.2-251.4℃; white powder solid. 1HNMR(400MHz,CDCl3)δ8.09(d,J=8.5Hz,1H,phenyl),7.56(dtd,J=8.2,7.0,1.0Hz,1H,phenyl),7.44(dtd,J=8.2,7.0,1.0Hz,1H,phenyl),7.34(d,J=8.3Hz,1H,phenyl),5.71(s,1H,CH-12),3.22(dd,J=10.7,5.6Hz,1H,CH-3),2.78(dt,J=13.5,3.6Hz,1H,CH-1),2.34(s,1H,CH-9),2.29-2.25,2.18-2.12,2.07,1.94-1.83,1.72-1.59,1.55-1.50,1.46,1.25(18H,methylene and methineof triterpenoid structure),1.58(s,3H,CH3-29),1.41(s,3H,CH3-27),1.16(s,3H,CH3-25),1.13(s,3H,CH3-26),1.01(s,3H,CH3-23),0.94(s,3H,CH3-24),0.81(s,3H,CH3-28),0.71(dd,1H,CH-5);13C NMR(101MHz,CDCl3)δ199.90(C-11),172.54(C-30),167.56(C-13),143.61(phenyl),129.10(phenyl),128.85(phenyl),128.62(C-12),124.83(phenyl),120.73(phenyl),107.84(phenyl),78.77(C-3),61.89(C-9),54.97(C-5),48.25(C-18),45.41(C-14),44.40(C-20),43.20(C-8),40.93(C-19),39.15(C-1),39.13(C-4),37.80(C-22),37.13(C-10),32.79(C-7),32.02(C-17),31.22(C-21),28.57(C-29),28.12(C-28),28.06(C-23),27.31(C-2),26.40(C-15 / C-16,2C),23.52(C-27),18.72(C-26),17.50(C-6),16.37(C-25),15.59(C-24);IRνmax 3116,1791,1652,1463,1384,1110,1068,1049,866,750cm. -1 .

[0054] Example 5

[0055]

[0056] Weigh 2.00 mmol of L-methionine into a 50 mL three-necked reaction flask, add 40 mL of acetonitrile to dissolve it, then weigh 0.40 mmol of the compound shown in Formula 4 and add 5 mL of dry dichloromethane solution to dissolve it. A dichloromethane solution of the compound shown in Formula 4 was added to an acetonitrile solution of L-methionine, and 4.00 mmol of triethylamine was added dropwise. The reaction was carried out by stirring at room temperature, and the reaction system was monitored by thin-layer chromatography at regular intervals. Stirring was stopped when a large amount of the target product was generated and 18β-GA no longer decreased. The reaction system was then poured into 15 mL of ice water to terminate the reaction. The mixture was transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane. The organic phase was collected and then extracted twice with saturated sodium chloride aqueous solution to remove water-soluble impurities. The organic phase was collected and then dried with anhydrous sodium sulfate to remove excess water. After filtration and concentration under reduced pressure, the target crude product was obtained. After purification by recrystallization, the product was concentrated under reduced pressure to obtain the compound shown in Formula 5. The physical properties and characterization data of the obtained compound are as follows: yield: 40%; melting point: 225.2-226.5℃; white powder solid. 1 H NMR(400MHz,CDCl3)δ6.98(s,1H,CONH),5.79(s,1H,CH-12),4.60(s,H,NHCH),3.23(dd,J=10.4,4.8Hz,1H,CH-3),3.19-3.12(m,2H,CH2SCH3),2 .73(d,J=12.9Hz,1H,CH-1),2.55-2.42(m,2H,CHCH2),2.32(s,1H,CH-9),2.06(s,3H,SCH3),2.21,1.80,1.61,1.47-1.38,1.27(18H,methylene and methine of triterpenoid structure),1.36(s,3H,CH3-29),1.15(s,3H,CH3-25),1.11(s,3H,CH3-26),1.10(s,3H, CH3-27),0.99(s,3H,CH3-23),0.79(s,6H,CH3-24,CH3-28),0.68(d,J=11.4Hz,1H,CH-5);13 C NMR(101MHz, CDCl3)δ201.35(C-11),176.18(COOH),174.63(C-30),170.96(C-13),127.70(C-12),78.86(C-3),61.75(C-9),54. 94(C-5),51.21(NHCH),48.15(C-18),45.70(C-8),43.79(C-14),43.25(C-19),41.57(C-4),39.15(C-1 / C-10),37.30(C-20),36 .98(C-22),32.79(C-21),31.93(C-7),31.56(C-17),31.19(CHCH2),30.17(CH2SCH3),29.23(C-2),28.42(C-28),28.13(C-29), 27.04(C-16),26.56(C-15),26.30(C-27),23.24(C-23 / C-24,2C),18.62(C-6),17.45(C-26),16.26(C-25),15.66(SCH3); IRνmax 2951,2926,1712,1656,1517,1467,1386,1207,1087,1037,921cm -1 .

[0057] Example 6

[0058]

[0059] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of 3-phenylpropionic acid and add it to the above mixture to dissolve. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with petroleum ether:ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 6. The physical properties and characterization data of the obtained compound are as follows: yield: 52%; melting point: 130.0-132.0℃; white powder solid.1 HNMR(400MHz,CDCl3)δ7.29(d,J=8.0Hz,2H,H-Ar),7.22-7.14(m,3H,H-Ar),5.67(s,1H,CH-12),4.52(dd,J=11.6,4.7Hz,1H,CH-3),3.72(s,3H,COOCH3),3.67-3.57(m,4H,N(CH2)2),3.50-3.42(t,4H,N(CH2)2),2.96(t,J=7.6Hz,2H,Ph-CH2),2.78(dd,J=10.6,3.1Hz,1H,CH-1),2.64(t,J=7.8Hz,2H,CH2COO),2.35(s,1H,CH-9),2.29,2.06,2.01-1.95,1.88-1.78,1.64,1.58,1.55,1.42,1.01(18H,methylene and methine of triterpenoid structure),1.35(s,3H,CH3-29),1.23(s,3H,CH3-25),1.15(s,3H,CH3-26),1.11(s,3H,CH3-27),1.04(d,J=2.4Hz,1H,CH-5),0.85(s,3H,CH3-23),0.81(s,3H,CH3-24),0.80(s,3H,CH3-28); 13 C NMR(101MHz,CDCl3)δ199.98(C-11),174.25(Phe-COO),172.76(C-13),169.46(C-30),155.83( C OOCH3),140.53(C-Ar),128.57(C-12),128.46(C-Ar,2C),128.28(C-Ar,2C),126.22(C-Ar),80.70(C-3),61.71(C-9),55.04(C-5),52.85(COO C H3),48.15(C-18),45.31(N(CH2)2,2C),43.93(N(CH2)2,2C),43.84(C-8),43.30(C-14),38.80(C-19),38.06(C-4),37.73(C-1),36.95(C-10),36.24(C-20),35.34(C-21),33.21(Phe-CH2 CH2),32.76(C-22),31.79(C-7),31.11(C-17),30.72(Phe- C H2),28.43(C-2),27.99(C-28),27.08(C-29),26.73(C-16),26.42(C-15),23.54(C -27),23.12(C-23),18.70(C-24),17.38(C-6),16.73(C-26),16.41(C-25); IRνmax 2951,2926,1708,1656,1633,1465,1448,1251,1213,1072,1028,986,975,759,736cm -1 HRMS(ESI)m / z calcd for C 56 H 64 N3O8SNa + (M+Na) + 972.51671, found 972.51654.

[0060] Example 7

[0061]

[0062] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Fmoc-L-methionine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with petroleum ether:ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 7. The physical properties and characterization data of the obtained compound are as follows: yield: 52%; melting point: 130.0-132.0℃; white powder solid. 1H NMR(400MHz,CDCl3)δ7.77(d,J=7.5Hz,2H,H-Ar),7.60(d,J=7.5Hz,2H,H-Ar),7.40(t,J=7.4Hz,2H,H-Ar),7.32(dd,J=10.8,4.5Hz,2H,H-Ar),5.68(s,1H,CH-12),5.44(dd,J=14.5,8.2Hz,1H,CONH),4.58(dd,J=11.7,4.7Hz,1H,CH-3),4.50(t,J=6.7Hz,1H,CHNH),4.41(dd,J=6.7,3.2Hz,2H,Fmoc-L-Met-COOCH2),4.23(t,J=7.0Hz,1H,Fmoc-Cyclopentane-CH),3.72(s,3H,COOCH3),3.67-3.57(m,4H,N(CH2)2),3.47(t,J=5.3Hz,4H,N(CH2)2),2.82(d,J=13.6Hz,1H,CH-1),2.54(s,2H,CH2SCH3),2.35(s,1H,CH-9),2.10(s,3H,SCH3),2.07-2.01(m,2H,CH2CH2SCH3),2.30,2.01-1.95,1.81,1.70-1.60,1.56,1.47-1.39,1.03(19H,methylene and methine oftriterpenoid structure),1.36(s,3H,CH3-29),1.22(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-27),0.88(s,6H,CH3-23 / CH3-24,),0.81(s,3H,CH3-28); 13¹³C NMR (101 MHz, CDCl₃) δ 199.80 (C-11), 174.20 (Fmoc-L-Met-CO), 171.67 (C-13), 169.50 (C-30), 155.95 (Fmoc-L-Met-NHCO), 155.81 (COOCH₃), 143.88 (Ar-C), 143.74 (Ar-C), 141.33 (Ar-C, 2C), 128.56 (C-12), 127.74 (Ar-C, 2C), 127.08 (Ar-C, 2C), 125.10 (Ar-C), 125.08 (Ar-C), 120.01 (Ar-C), 119.99 (Ar-C), 82.49 (C-3), 67.02 (Fmoc-L-Met-COOCH₂), 61.65 (C-9), 55.03 (C-5), 53.66 (CHNH), 52.85 (COOCH₃), 48.11 (C-18), 47.19 (Fmoc-Cyclopentane-CH), 45.29 (N(CH₂)₂, 2C), 43.93 (N(CH₂)₂, 2C), 43.89 (C-8), 43.31 (C-14), 38.72 (C-19), 38.17 (C-4), 37.73 (C-20), 36.94 (C-1 / C-10), 33.14 (C-21), 32.73 (C-22), 32.27 (C-7), 31.79 (CH₂CH₂SCH₃), 29.99 (CH₂CH₂SCH₃), 28.43 (C-17), 28.22 (C-2), 27.08 (C-28), 26.72 (C-29), 26.42 (C-16), 23.58 (C-15), 23.14 (C-27), 18.70 (C-23 / C-24, 2C), 17.37 (C-6), 16.84 (C-26), 16.39 (C-25), 15.49 (SCH₃); IR νmax 2951, 2926, 1708, 1656, 1633, 1465, 1448, 1251, 1213, 1072, 1028, 986, 975, 759, 736 cm -1 ; HRMS (ESI) m / z calcd for C 56 H 75 N₃O₈SNa + (M+Na) + 972.51671, found 972.51654.

[0063] Example 8

[0064]

[0065] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Boc-glycine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with petroleum ether:ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 8. The physical properties and characterization data of the obtained compound are as follows: yield: 78%; melting point: 252.2-253.0℃; white powder solid. 1 H NMR (400MHz, CDCl3) δ5.68 (s, 1H, CH-12), 5.02 (d, J = 6.2Hz, 1H, Gly-CH2NH), 4.59 (dd, J = 11.7, 4. 7Hz,1H,CH-3),3.90(d,J=5.3Hz,2H,Gly-CH2NH),3.72(s,3H,COOCH3),3.62(m,4H,N(CH2)2),3.4 7(t,J=5.2Hz,4H,N(CH2)2),2.81(dt,J=13.6,3.7Hz,1H,CH-1),2.35(s,1H,CH-9),2.30,2.10-2 .01,2.01-1.95,1.85,1.76-1.60,1.57,1.43-1.42,1.41-1.38,1.07-0.99,0.85(19H,methylene and methine of triterpenoid structure),1.45(s,9H,Boc-CH3),1.36(s,3H,CH3-29),1.23(s,3H,CH3-25),1.16(s ,3H,CH3-26),1.12(s,3H,CH3-27),0.88(s,6H,CH3-24 / CH3-23),0.81(s,3H,CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.84 (C-11), 174.19 (Gly-COO), 170.14 (C-13), 169.47 (C-30), 155.79 (Boc-COO), 155.65 (NCOO), 128.54 (C-12), 81.94 (C-3), 79.84 (Boc-q.C), 61.66 (C-9), 55.02 (C-5), 52.82 (OCH3), 48.11 (C-18), 45.28 (N(CH2)2, 2C), 43.91 (N(CH2)2, 2C), 43.86 (C-8), 43.29 (C-14), 42.66 (Gly-CHNH), 38.74 (C-19), 38.14 (C-4), 37.72 (C-1), 36.92 (C-10), 33.14 (C-20), 32.73 (C-21), 31.77 (C7 / C-22), 28.42 (C-17), 28.33 (Boc-CH3, 3C), 28.08 (C-2), 27.06 (C-28), 26.70 (C-29), 26.40 (C-16), 23.53 (C-15), 23.12 (C-27), 18.68 (C-23 / C-24, 2C), 17.36 (C-6), 16.66 (C-26), 16.41 (C-25); IR νmax 2972, 2929, 1708, 1656, 1633, 1465, 1448, 1251, 1213, 1072, 1028, 986, 975, 759, 736 cm -1 -1; HRMS (ESI) m / z calcd for C 43 42H 67 63N3O8Na + + (M+Na) + 776.48204, found 776.48181.

[0066] Example 9

[0067]

[0068] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Boc-L-leucine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (eluting with petroleum ether:ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 9. The physical properties and characterization data of the obtained compound are as follows: yield: 50%; melting point: 249.9-252.2℃; white powder solid. 1 H NMR (400MHz, CDCl3) δ5.68 (s, 1H, CH-12), 5.01 (d, J = 8.9Hz, 1H, Leu-CHNH), 4.56 (dd, J = 11.4, 4.8 Hz,1H,CH-3),4.26(dd,J=8.7,3.8Hz,1H,Leu-CHNH),3.72(s,3H,COOCH3),3.67-3.58(m,4H,N(CH 2)2),3.51-3.43(m,4H,N(CH2)2),2.80(dt,J=13.7,3.7Hz,1H,CH-1),2.35(s,1H,CH-9),2.29,2 .07-2.02,1.98,1.88-1.82,1.76-1.70,1.68-1.59,1.57,1.40,1.02,0.92,0.86(22H,methylene and methine of triterpenoid structure and leucine),1.44(s,9H,Boc-CH3),1.36(s,3H,CH3-29),1.23(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s, 3H,CH3-27),0.98-0.94(m,6H,Leu-CH3),0.90(s,3H,CH3-24),0.88(s,3H,CH3-23),0.81(s,3H,CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.84 (C-11), 174.20 (Ile-COO), 172.01 (C-13), 169.42 (C-30), 155.79 (Boc-COO), 155.59 (COOCH3), 128.56 (C-12), 81.91 (C-3), 79.61 (Boc-q.C), 61.66 (C-9), 58.51 (Leu-CHNH), 55.07 (C-5), 52.83 (COOCH3), 48.13 (C-18), 45.29 (N(CH2)2, 2C), 43.91 (N(CH2)2, 2C), 43.85 (C-8), 43.30 (C-14), 38.74 (C-19), 38.04 (C-4), 37.97 (Leu-CH), 37.72 (C-1), 36.92 (C-10), 33.19 (C-20), 32.75 (C-21), 31.78 (C-7 / C-22), 28.42 (C-17), 28.36 (Boc-CH3, 3C), 28.08 (C-2), 27.08 (C-28), 26.72 (C-29), 26.41 (C-16), 24.73 (Leu-CH2), 23.62 (C-15), 23.12 (C-27), 18.70 (C-23 / C-24, 2C), 17.37 (C-6), 16.86 (C-26), 16.37 (C-25), 15.74 (Ile-CH3CHCH3), 11.68 (Leu-CH3CHCH3); IR νmax 2976, 2870, 1710, 1654, 1631, 1529, 1465, 1415, 1392, 1284, 1120, 1010, 981, 779 cm -1 ; HRMS (ESI) m / z calcd for 47 H 75 N3O8Na + (M+Na) + 832.54464, found 832.54437.

[0069] Example 10

[0070]

[0071] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Boc-L-methionine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with petroleum ether:ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 10. The physical properties and characterization data of the obtained compound are as follows: yield: 80%; melting point: 249.8-250.5℃; white powder solid. 1 H NMR(400MHz, CDCl3) δ5.68(s,1H,CH-12),5.11(d,J=8.0Hz,1H,Met-CHNH),4.57(dd,J=11.6,4.5Hz,1H,CH-3),4 .44-4.36(m,1H,Met-CHNH),3.72(s,3H,COOCH3),3.67-3.56(m,4H,N(CH2)2),3.47(t,J=5.2Hz,4H,N(CH2)2),2 .82(dt,J=13.6Hz,1H,CH-1),2.58-2.53(m,2H,Met-CH2S),2.35(s,1H,CH-9),2.10(s,3H,Met-CH3),2.08-2.03 (m,2H,Met-CH2CH),2.29,2.03-1.95,1.91-1.85,1.82,1.73,1.70-1.60,1.57,1.40,1.03,0.85(19H,methylene andmethine of triterpenoid structure),1.45(s,9H,Boc-CH3),1.36(s,3H,CH3-29),1.23(s,3H,CH3-25),1.16(s,3H,CH 3-26),1.12(s,3H,CH3-27),0.90(s,3H,CH3-23),0.88(s,3H,CH3-24),0.81(s,3H,CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.81 (C-11), 174.19 (Met-COO), 171.99 (C-13), 169.47 (C-30), 155.79 (Boc-COO), 155.30 (COOCH3), 128.55 (C-12), 82.17 (C-3), 79.90 (Boc-q.C), 61.65 (C-9), 55.03 (C-5), 53.26 (Met-CHNH), 52.83 (COOCH3), 48.11 (C-18), 45.28 (N(CH2)2, 2C), 43.92 (N(CH2)2, 2C), 43.86 (C-8), 43.30 (C-14), 38.71 (C-19), 38.14 (C-4), 37.72 (C-1), 36.92 (C-10), 33.14 (C-20), 32.73 (C-21), 32.47 (Met-CH2CH), 31.78 (C-22), 30.03 (C-7), 29.93 (Met-CH2S), 28.42 (C-17), 28.33 (Boc-CH3, 3C), 28.19 (C-2), 27.07 (C-28), 26.91 (C-29), 26.71 (C-16), 26.41 (C-15), 23.55 (C-27), 23.12 (C-23), 18.69 (C-24), 17.36 (C-6), 16.82 (C-26), 16.39 (C-25), 15.48 (Met-CH3); IR νmax 2953, 2947, 1712, 1656, 1633, 1448, 1390, 1253, 1238, 1168, 1093, 985, 765 cm -1 ; HRMS (ESI) m / z calcd for C 46 H 73 N3O8SNa + (M+Na) + 850.50106, found 850.50067.

[0072] Example 11

[0073]

[0074] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Boc-L-isoleucine and add it to the above mixture to dissolve. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with petroleum ether:ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 11. The physical properties and characterization data of the obtained compound are as follows: yield: 70%; melting point: 150.8-152.0℃; white powder solid. 1 H NMR (400MHz, CDCl3) δ5.68 (s, 1H, CH-12), 5.01 (d, J = 8.9Hz, 1H, Ile-CHNH), 4.56 (dd, J = 11.4, 4.8 Hz,1H,CH-3),4.26(dd,J=8.7,3.8Hz,1H,Ile-CHNH),3.72(s,3H,COOCH3),3.67-3.58(m,4H,N(CH 2)2),3.51-3.43(m,4H,N(CH2)2),2.80(dt,J=13.7,3.7Hz,1H,CH-1),2.35(s,1H,CH-9),2.29,2 .07-2.02,1.98,1.88-1.82,1.76-1.70,1.68-1.59,1.57,1.40,1.02,0.92,0.86(22H,methylene and methine oftriterpenoid structure and isoleucine),1.44(s,9H,Boc-CH3),1.36(s,3H,CH3-29),1.23(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12( s,3H,CH3-27),0.98-0.94(m,6H,Ile-CH3),0.90(s,3H,CH3-24),0.88(s,3H,CH3-23),0.81(s,3H,CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.84 (C-11), 174.20 (Ile-COO), 172.01 (C-13), 169.42 (C-30), 155.79 (Boc-COO), 155.59 (COOCH3), 128.56 (C-12), 81.91 (C-3), 79.61 (Boc-q.C), 61.66 (C-9), 58.51 (Ile-CHNH), 55.07 (C-5), 52.83 (COOCH3), 48.13 (C-18), 45.29 (N(CH2)2, 2C), 43.91 (N(CH2)2, 2C), 43.85 (C-8), 43.30 (C-14), 38.74 (C-19), 38.04 (C-4), 37.97 (Ile-CH), 37.72 (C-1), 36.92 (C-10), 33.19 (C-20), 32.75 (C-21), 31.78 (C-7 / C-22), 28.42 (C-17), 28.36 (Boc-CH3, 3C), 28.08 (C-2), 27.08 (C-28), 26.72 (C-29), 26.41 (C-16), 24.73 (Ile-CH2), 23.62 (C-15), 23.12 (C-27), 18.70 (C-23 / C-24, 2C), 17.37 (C-6), 16.86 (C-26), 16.37 (C-25), 15.74 (Ile-CHCH3), 11.68 (Ile-CH2CH3); IR νmax 2960, 2929, 1726, 1658, 1627, 1492, 1390, 1286, 1251, 1238, 1213, 1168, 1012, 983, 785 cm -1 ; HRMS (ESI) m / z calcd for C 47 H 75 N3O8Na + (M+Na) + 832.54464, found 832.54437.

[0075] Example 12

[0076]

[0077] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Boc-L-phenylalanine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with petroleum ether:ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 12. The physical properties and characterization data of the obtained compound are as follows: yield: 82%; melting point: 241.1-242.5℃; white powder solid. 1 H NMR(400MHz, CDCl3)δ7.30(d,J=6.8Hz,1H,H-Ar),7.27-7.19(m,2H,H-Ar),7.17(t,J=6.9Hz,2H ,H-Ar),5.68(s,1H,CH-12),4.96(dd,J=49.1,8.1Hz,1H,Phe-CHNH),4.62-4.42(m,2H,CH-3and Phe-CHNH),3.72(s,3H,COOCH3),3.68-3.58(m,4H,N(CH2)2),3.47(t,J=5.3Hz,4H,N(CH2)2),3.15-2.99(m,2H,Phe-CH2),2.79(d,J =13.4Hz,1H,CH-1),2.34(s,1H,CH-9),2.30,2.10-2.02,1.98,1.90-1.76,1.71,1.69-1.50,1.47,1.27,1.02,0.83(19H,methylene and methine of triterpenoid structure),1.43(s,3H,CH3-29),1.40(s,9H,Boc-CH3),1.35(s,3H,CH3-25),1.23(s ,3H,CH3-26),1.14(s,3H,CH3-27),1.11(s,3H,CH3-23),0.83(s,6H,CH3-24 / CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.85 (C-11), 174.20 (Phe-COO), 171.80 (C-13), 169.44 (C-30), 155.80 (Boc-COO), 155.06 (COOCH3), 136.11 (Phe-C), 129.38 (Phe-C, 3C), 128.56 (Phe-C / C-12, 2C), 126.97 (Phe-C), 82.16 (C-3), 79.79 (Boc-q.C), 61.67 (C-9), 55.05 (C-5), 54.67 (Phe-CHNH), 52.83 (COOCH3), 48.12 (C-18), 45.28 (N(CH2)2, 2C), 43.92 (N(CH2)2, 2C), 43.87 (C-8), 43.29 (C-14), 38.74 (C-19), 38.46 (Phe-CH2), 38.06 (C-4), 37.73 (C-1), 36.89 (C-10), 33.16 (C-20), 32.74 (C-21), 31.78 (C-22 / C-7, 2C), 28.42 (C-17), 28.32 (Boc-CH3, 3C), 28.04 (C-2), 27.08 (C-28), 26.91 (C-29), 26.72 (C-16), 26.41 (C-15), 23.44 (C-27), 23.12 (C-23), 18.69 (C-24), 17.33 (C-6), 16.70 (C-26), 16.38 (C-25); IR νmax 2972, 2953, 1712, 1656, 1635, 1465, 1450, 1390, 1284, 1170, 1078, 1049, 700 cm -1 ; HRMS (ESI) m / z calcd for C 50 H 73 N3O8Na + (M+Na) + 866.52899, found 866.52881.

[0078] Example 13

[0079]

[0080] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Boc-L-proline into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with petroleum ether:ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 13. The physical properties and characterization data of the obtained compound are as follows: yield: 76%; melting point: 253.7-254.8℃; white powder solid. 1 H NMR (400MHz, CDCl3) δ5.68 (d, J=5.8Hz, 1H, CH-12), 4.54 (m, 1H, CH-3), 4.28 (m, 1H, Pro-CH), 3 .72(s,3H,COOCH3),3.69-3.59(m,4H,N(CH2)2),3.57-3.51(m,2H,Pro-CH2),3.47(t,J=5.3Hz ,4H,N(CH2)2),2.80(dt,J=12.3,6.7Hz,1H,CH-1),2.35(s,1H,CH-9),2.20(m,1H,Pro-CHH') ,2.28,2.11-1.95,1.92-1.82,1.72,1.70-1.60,1.57,1.40,1.03,0.88,0.86(22H,methylene and methine of triterpenoid structure and proline),1.46(s,3H,CH3-29),1.42(s,9H,Boc-CH3),1.36(s,3H,CH3-25),1.23(s,3H,CH3 -26),1.16(s,3H,CH3-27),1.12(s,3H,CH3-24),0.89(s,3H,CH3-23),0.81(s,3H,CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.86 (C-11), 174.20 (Pro-COO), 172.84 (C-13), 169.48 (C-30), 155.79 (Boc-COO), 153.89 (COOCH3), 128.54 (C-12), 81.08 (C-3), 79.92 (Boc-q.C), 61.67 (C-9), 59.41 (Pro-CH), 55.03 (C-5), 52.82 (COOCH3), 48.10 (C-18), 46.45 (C-8), 46.27 (Pro-CH2), 45.28 (N(CH2)2, 2C), 43.91 (N(CH2)2, 2C), 43.29 (C-14), 38.71 (C-19), 38.13 (C-4), 37.72 (C-1), 36.92 (C-10), 33.11 (C-20), 32.73 (C-21), 31.77 (C-7 / C-22, 2C), 28.50 (Pro-CH2), 28.42 (C-17), 28.38 (Boc-CH3, 3C), 28.20 (C-2), 27.07 (C-28), 26.71 (C-29), 26.41 (C-16), 24.32 (Pro-CH2), 23.49 (C-15), 23.11 (C-27), 18.69 (C-23 / C-24), 17.35 (C-6), 16.72 (C-26), 16.39 (C-25); IR νmax 2980, 2929, 1703, 1654, 1627, 1469, 1413, 1392, 1282, 1255, 1176, 1165, 985, 767 cm -1 ; HRMS (ESI) m / z calcd for C 46 H 71 N3O8Na + (M+Na) + 816.51334, found 816.51318.

[0081] Example 14

[0082]

[0083] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Boc-N-methyl-L-alanine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with petroleum ether:ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 14. The physical properties and characterization data of the obtained compound are as follows: yield: 80%; melting point: 249.8-252.4℃; white powder solid. 1 H NMR(400MHz, CDCl3)δ5.68(s,1H,CH-12),4.62-4.46(m,2H,CH-3 / N-Methyl-Ala-C H),3.72(s,3H,COOCH3),3.67-3.58(m,4H,N(CH2)2),3.48(t,J=5.2Hz,4H,N(CH2)2 ),2.86(d,J=5.4Hz,1H,CH-1),2.83-2.77(br,3H,N-Methyl-Ala-NCH3),2.35(s,1H ,CH-9),2.30,2.10-1.95,1.89-1.82,1.77,1.72-1.54,1.40,1.03(19H,methylene and methhine of triterpenoidstructure),1.46(s,9H,Boc-CH3),1.45(s,3H,N-Methyl-Ala-CHCH3),1.36(s,3H,CH3-29),1.2 3(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-27),0.87(s,6H,CH3-23 / CH3-24),0.81(s,3H,CH3-28); 13¹³C NMR (101 MHz, CDCl₃) δ 199.88 (C-11), 174.20 (N-Methyl-Ala-COO), 172.18 (C-13), 169.46 (C-30), 155.78 (Boc-COO / COOCH₃, 2C), 128.54 (C-12), 81.45 (C-3), 79.81 (Boc-q.C), 61.66 (C-9), 54.98 (N-Methyl-Ala-CHCH₃), 53.64 (C-5), 52.82 (COOCH₃), 48.11 (C-18), 45.28 (N(CH₂)₂, 2C), 43.91 (N(CH₂)₂, 2C), 43.86 (C-8), 43.29 (C-14), 38.70 (C-19), 38.07 (C-4), 37.72 (C-1), 36.92 (C-10), 33.14 (C-20), 32.73 (C-21), 31.77 (C-7 / C-22, 2C), 30.37 (N-Methyl-Ala-NCH₃), 28.42 (C-17), 28.40 (Boc-CH₃, 3C), 28.08 (C-2), 27.06 (C-28), 26.70 (C-29), 26.40 (C-16), 23.47 (C-15), 23.12 (C-27), 18.68 (C-23 / C-24), 17.36 (C-6), 16.75 (C-26), 16.40 (C-25), 14.90 (N-Methyl-Ala-CHCH₃); IR νmax 2983, 2926, 1726, 1691, 1629, 1462, 1411, 1390, 1284, 1257, 1211, 1178, 1149, 1070, 1012, 867, 767 cm -1 ; HRMS (ESI) m / z calcd for C 45 H 71 N₃O₈Na + (M+Na) + 804.51334, found 804.51306.

[0084] Example 15

[0085]

[0086] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve them, and then weigh 1.20 mmol of Boc-L-valine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with petroleum ether: ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 15. The physical properties and characterization data of the obtained compound are as follows: yield: 78%; melting point: 107.6-109.1℃; white solid powder. 1 H NMR (400MHz, CDCl3) δ5.68 (s, 1H, CH-12), 5.00 (d, J = 9.3Hz, 1H, Boc-L-valine CHNH),4.56(dd,J=11.8,4.8Hz,1H,CH-3),3.72(s,3H,COOCH3),3.67-3.52(m,4H ,N(CH2)2),3.46(d,J=5.5Hz,4H,N(CH2)2),2.81(d,J=13.6Hz,1H,Boc-L-valine CHNH),2.35(s,1H,CH-9),2.32-2.24(m,1H,Boc-L-valine CHCH3),2.19(q,J=6.0Hz,1H,CH-18),2.05,1.65,1.51,1.40,1.27,0.86,0.79(19H,methylene and methine oftriterpenoid structure),1.45(s,9H,Boc-CH3),1.36(s,3H,CH-25),1.23(s,3H,CH-26),1.16(s,3H,CH-27),1 .12(s,3H,CH-28),0.99(d,J=7.0Hz,6H,CH-23 / CH-24),0.89(d,J=4.9Hz,9H,CH-29 / Boc-L-valine CHCH3); 13C NMR (101 MHz, CDCl3) δ 199.87 (C-11), 174.22 (L-valine-COO), 172.10 (C-13), 169.46 (C-30), 155.81 (COOCH3), 155.76 (Boc-COO), 128.57 (C-12), 81.88 (C-3), 79.63 (Boc-q.C), 61.67 (C-9), 58.91 (NHCH), 55.06 (C-5), 52.83 (COOCH3), 48.13 (C-18), 45.29 (N(CH2)2, 2C), 43.92 (N(CH2)2, 2C), 43.84 (C-8), 43.30 (C-14), 38.74 (C-19), 38.07 (C-4), 37.72 (C-1), 36.92 (C-10), 33.19 (C-20), 32.75 (C-21), 31.78 (C-7 / C-22, 2C), 31.19 (Boc-L-valine CH(CH3)2), 28.42 (C-17), 28.35 (Boc-CH3, 3C), 28.12 (C-2), 27.07 (C-28), 26.71 (C-29), 26.42 (C-16), 23.64 (C-15), 23.12 (C-27), 19.33 (Boc-L-valine CH(CH3)2, 2C), 18.70 (C-23), 17.21 (C-24), 17.21 (C-6), 16.84 (C-26), 16.37 (C-25); IR vmax 2970, 2358, 2339, 1712, 1620, 1510, 1467, 1249, 1171, 1010, 765 cm -1 ; HRMS (ESI) m / z calcd for C 46 H 73 N3O8Na + (M+Na) + 818.52899, found 818.52875.

[0087] Example 16

[0088]

[0089] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve them, and then weigh 1.20 mmol of Boc-L-aspartic acid-4-benzyl ester into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with petroleum ether:ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 16. The physical properties and characterization data of the obtained compound are as follows: yield: 42%; melting point: 152.6-154.5℃; white solid powder. 1 HNMR(400MHz, CDCl3)δ7.40-7.31(m,5H,H-Ar),5.68(s,1H,CH-12),5.52(d,J=8.9Hz,1H,CHNH),5 .17(s,1H,CHNH),5.16-5.06(m,2H,Ar-CH2),4.56(s,1H,CH-3),3.72(s,3H,COOCH3),3.69-3.53(m ,4H,N(CH2)2),3.47(s,4H,N(CH2)2),2.87(dd,J=17.1,4.7Hz,2H,CH2COO),2.34(s,1H,CH-9),2. 04(s,1H,CH-18),2.29,1.96,1.83,1.56-1.48,1.40,1.30,1.10-0.91,0.87,0.76(19H,methylene and methine oftriterpenoid structure),1.64(s,3H,CH-25),1.44(s,9H,Boc-CH3),1.36(s,3H,CH-26),1.23(s,3H, CH-27),1.15(s,3H,CH-28),1.12(s,3H,CH-29),0.83(s,3H,CH-23),0.81(s,3H,CH-24); 1313C NMR (101 MHz, CDCl3) δ 199.87 (C-11), 174.23 (COO, 2C), 170.98 (CONH), 170.78 (C-13), 169.46 (C-30), 155.81 (Boc-COO), 155.46 (COOCH3), 135.37 (Ar-C), 128.62 (Ar-C), 128.57 (Ar-C / C-12, 2C), 128.51 (Ar-C), 128.42 (Ar-C), 128.38 (Ar-C), 82.48 (C-3), 79.97 (Boc-q.C), 66.83 (Ar-CH2), 61.66 (C-9), 55.04 (COOCH3), 52.83 (NHCH), 50.20 (C-5), 48.14 (C-18), 45.29 (N(CH2)2, 2C), 43.92 (N(CH2)2, 2C), 43.86 (C-8), 43.30 (C-14), 38.70 (C-19), 38.07 (C-4), 37.73 (C-1), 36.91 (C-10), 36.74 (CH2), 33.18 (C-20), 32.74 (C-21), 31.79 (C-22 / C-7, 2C), 28.43 (C-17), 28.33 (Boc-CH3, 3C), 28.03 (C-2), 27.89 (C-29), 27.08 (C-28), 26.72 (C-16), 26.42 (C-15), 23.38 (C-27), 23.13 (C-23), 18.69 (C-24), 17.33 (C-6), 16.70 (C-26), 16.37 (C-25); IR vmax 3442, 2972, 2870, 1710, 1649, 1620, 1510, 1467, 1390, 1365, 1289, 1236, 1166, 1010, 748, 698 cm -1 ; HRMS (ESI) m / z calcd for C 52 H 75 N3O 10 Na + (M+Na) + 924.53447, found 924.53491.

[0090] Example 17

[0091]

[0092] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve them, and then weigh 1.20 mmol of Boc-L-tryptophan into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored by thin-layer chromatography at regular intervals. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane and then twice with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was separated and purified by silica gel column chromatography (elution with petroleum ether: ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 17. The physical properties and characterization data of the obtained compound are as follows: Yield: 62%, yellow solid powder; 1 H NMR (400MHz, CDCl3) δ8.29(s,1H,C=C-NH),7.60(d,J=7.8Hz,1H,H-Ar),7.34(d,J=8.2Hz,1H,H-Ar),7.27(s,2H,H-Ar),7.13- 7.09(m,1H,H-Ar),6.99(d,J=2.3Hz,1H,CH=C),5.68(s,1H,CH-12),5.10(d,J=8.2Hz,1H,CHNH),4.66(q,J=7.1,6.5Hz,1H,CH- 3),4.46(dd,J=10.4,6.1Hz,1H,CHNH),3.72(s,3H,COOCH3),3.61(dt,J=11.2,4.9Hz,4H,N(CH2)2),3.46(t,J=5.1Hz,4H,N(CH 2)2),2.32(s,1H,CH-9),2.04(s,1H,CH-18),2.01-1.79,1.59-1.46,1.40,1.29,1.26,1.16,1.01,0.78,0.72(19H,methylene and methine oftriterpenoid structure),1.69-1.59(m,3H,CH-25),1.43(s,9H,Boc-CH3),1.34(s,3H,CH-26),1.22(s ,3H,CH-27),1.11(d,J=3.0Hz,6H,CH-28 / CH-29),0.81(s,3H,CH-23),0.77(s,3H,CH-24);13 C NMR (101 MHz, CDCl3) δ 200.03 (C-11), 174.28 (COO, 2C), 172.17 (C-13), 169.67 (C-30), 155.85 (Boc-COO), 128.55 (NH-C=C / C-12, 2C), 127.89 (Ar-C), 122.81 (Ar-C), 122.10 (Ar-C), 119.49 (Ar-C), 119.06 (Ar-C), 111.14 (Ar-C), 110.35 (CH=C), 81.96 (C-3), 79.70 (Boc-q.C), 61.66 (C-9), 55.00 (COOCH3), 52.87 (NHCH), 48.12 (C-18), 45.30 (N(CH2)2, 2C), 43.94 (N(CH2)2, 2C), 43.89 (C-8), 43.31 (C-14), 38.70 (C-19), 38.02 (C-4), 37.73 (C-1), 36.89 (C-10), 33.11 (C-20), 32.71 (C-21), 31.79 (C-22 / C-7, 2C), 28.43 (C-17), 28.36 (Boc-CH3, 3C), 28.14 (C-2), 27.93 (C-29), 27.06 (C-28), 26.72 (C-16), 26.40 (C-15), 23.25 (C-27), 23.11 (C-23), 18.68 (C-24), 17.33 (C-6), 16.58 (C-25); IR vmax 3377, 2303, 1710, 1658, 1236, 1010, 985, 856, 767 cm -1 .

[0093] Example 18

[0094]

[0095] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve them, and then weigh 1.20 mmol of N-(tert-butoxycarbonyl)-N-methylglycine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored by thin-layer chromatography at regular intervals. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane and then twice with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then separated and purified by silica gel column chromatography (elution with petroleum ether: ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 18. The physical properties and characterization data of the obtained compound are as follows: Yield: 55%, white solid powder; 1 H NMR (400MHz, CDCl3) δ5.68 (d, J=3.1Hz, 1H, CH-12), 4.57 (ddd, J=11.1, 8.4, 4.9Hz, 1H, C H-3),4.00-3.82(m,2H,NCH2),3.72(s,3H,COOCH3),3.63(dq,J=15.7,8.0Hz,4H,N(CH2 )2),3.47(d,J=5.2Hz,4H,N(CH2)2),2.93(d,J=11.8Hz,3H,NCH3),2.04(s,1H,CH-18), 2.30,1.98,1.92-1.71,1.71-1.48,1.47,1.26,1.22-1.19,1.09-0.99(20H,methylene and methine oftriterpenoidstructure),1.43(s,9H,Boc-CH3),1.36(s,3H,CH3-29),1.23(s,3H,CH3-25),1.16(s ,3H,CH3-26),1.12(s,3H,CH3-27),0.88(dd,J=6.2,2.7Hz,6H,CH3-23 / CH3-24),0.81(s,3H,CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.87 (C-11), 174.22 (C-13), 169.79 (COO), 169.51 (C-30), 155.95 (Boc-COO), 155.81 (COOCH3), 128.56 (C-12), 81.59 (C-3), 80.12 (Boc-q.C), 61.67 (C-9), 55.03 (C-5), 52.82 (N(CH2)2, 2C), 51.31 (N(CH2)2, 2C), 50.73 (NCH2), 48.12 (C-18), 45.29 (NCH3), 43.91 (C-8), 43.87 (C-14), 43.29 (C-19), 38.74 (C-4), 38.08 (C-1), 37.72 (C-10), 36.92 (C-20), 33.14 (C-21), 32.73 (C-22), 31.77 (C-7), 28.42 (C-17), 28.36 (Boc-CH3, 3C), 28.31 (C-2), 28.11 (C-29), 27.06 (C-16), 26.71 (C-15), 26.40 (C-28), 23.60 (C-27), 23.12 (C-23), 18.68 (C-24), 17.37 (C-6), 16.75 (C-26), 16.41 (C-25); IR vmax 3159, 2927.94, 1656, 1408, 1323, 1211, 1049, 933 cm -1 ; HRMS (ESI) m / z calcd for C 44 H 69 N3O8Na + (M+Na) + 790.49769, found 790.49792.

[0096] Example 19

[0097]

[0098] Weigh 1.00 mmol of the compound shown in Formula 1, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve them, and then weigh 1.20 mmol of Boc-L-pyroglutamic acid into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored by thin-layer chromatography at regular intervals. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 1 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane and then twice with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was separated and purified by silica gel column chromatography (elution with petroleum ether: ethyl acetate = 10:1 to 1:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 19. The physical properties and characterization data of the obtained compound are as follows: Yield: 46%, white solid powder; 1 H NMR(400MHz,CDCl3)δ5.69(s,1H,CH-12),4.67-4.53(m,2H,CH-3 / NCH),3.72(s,3H,COOCH3),3.64(h,J=6.4Hz,4H,N(CH2)2),3.48(d,J=5.2Hz,4 H,N(CH2)2),2.54-2.27(m,4H,COCH2CH2),1.85(dd,1H,CH-18),1.80(d,1H,CH-9)2.08-1.99,1.76-1.52,1.26,1.07-1.04,1.02(19H,methylene and methineoftriterpenoid structure),1.50(s,9H,Boc-CH3),1.36(s,3H,CH3-29),1.23(s,3H,CH3-25),1.16(s,3H,CH 3-26),1.12(s,3H,CH3-27),0.89(s,3H,CH3-23),0.87(s,3H,CH3-24),0.81(s,3H,CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.80 (C-11), 174.21 (CON), 173.29 (C-13), 170.96 (COO), 169.60 (C-30), 155.80 (Boc-COO), 149.41 (COOCH3), 128.51 (C-12), 83.62 (C-3), 82.17 (Boc-q.C), 61.61 (C-9), 59.32 (COOCH3), 55.01 (C-5), 52.82 (N(CH2)2, 2C), 48.07 (C-18), 45.26 (N(CH2)2, 2C), 43.91 (CH2COO), 43.83 (C-8 / C-14, 2C), 43.29 (C-19), 38.63 (C-4), 38.12 (C-1), 37.71 (C-10), 36.89 (C-20), 33.04 (C-21), 32.68 (C-22), 31.76 (C-7), 31.19 (NCOCH2), 28.41 (C-17 / Boc-CH3, 4C), 28.16 (C-2), 27.95 (C-29), 27.04 (C-16), 26.68 (C-15), 26.39 (C-28), 23.53 (C-27), 23.10 (C-23), 21.97 (CH2), 18.67 (C-24), 17.32 (C-6), 16.72 (C-26), 16.36 (C-25); IR vmax 3294, 2976, 1708, 1627, 1456, 1321, 1217, 1070, 948 cm -1 ; HRMS (ESI) m / z calcd for C 46 H 69 N3O9Na + (M+Na) + 830.49260, found 830.49274.

[0099] Example 20

[0100]

[0101] Weigh 1.00 mmol of the compound shown in Formula 2, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve them, and then weigh 1.20 mmol of Boc-glycine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 2 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with dichloromethane:methanol = 20:1 to 12:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 20. The physical properties and characterization data of the obtained compound are as follows: yield: 72%; melting point: 231.3-233.3℃; white powder solid. 1 H NMR (400MHz, CDCl3) δ5.70 (s, 1H, CH-12), 5.02 (t, J = 5.8Hz, 1H, Gly-CH2NH), 4.57 (dd, J = 11.7 ,4.7Hz,1H,CH-3),3.89(d,J=5.6Hz,2H,Gly-CH2NH),3.65(t,J=5.1Hz,4H,N(CH2)2),2.89-2. 73(m,1H,CH-1),2.41(t,J=4.9Hz,4H,N(CH2)2),2.31(s,3H,NCH3),2.09(s,1H,CH-18),2.34 ,2.10-1.99,1.96,1.84,1.75-1.69,1.68-1.57,1.50,1.27,1.24,1.07-0.97(19H,methylene andmethine of triterpenoid structure),1.44(s,9H,Boc-CH3),1.34(s,3H,CH3-29),1.21(s,3H,CH3-25),1.14(s ,3H,CH3-26),1.11(s,3H,CH3-27),0.87(s,6H,CH3-23 / CH3-24),0.80(s,3H,CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.91 (C-11), 173.89 (Gly-COO), 170.16 (C-13), 169.68 (C-30), 155.65 (Boc-COO), 128.57 (C-12), 81.96 (C-3), 79.86 (Boc-q.C), 61.66 (C-9), 55.02 (C-5), 55.00 (N(CH2)2, 2C), 48.15 (C-18), 45.71 (NCH3), 45.29 (N(CH2)2, 2C), 43.87 (C-8), 43.81 (C-14), 43.29 (C-19), 42.67 (Gly-CHNH), 38.77 (C-4), 38.14 (C-1), 37.75 (C-10), 36.92 (C-20), 33.24 (C-21), 32.72 (C-22), 31.76 (C-7), 28.44 (C-17), 28.33 (Boc-CH3, 3C), 28.08 (C-2), 27.91 (C-29), 27.03 (C-16), 26.72 (C-15), 26.44 (C-28), 23.53 (C-27), 23.11 (C-23), 18.68 (C-24), 17.36 (C-6), 16.66 (C-26), 16.42 (C-25); IR νmax 2978, 2938, 2785, 1747, 1722, 1654, 1625, 1463, 1413, 1365, 1292, 1211, 1161, 1139, 974, 767 cm -1 ; HRMS (ESI) m / z calcd for C 42 H 68 N3O6 + (M+H) + 710.51026, found 710.51086.

[0102] Example 21

[0103]

[0104] Weigh 1.00 mmol of the compound shown in Formula 2, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Boc-L-leucine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 2 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with dichloromethane:methanol = 20:1 to 12:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 21. The physical properties and characterization data of the obtained compound are as follows: yield: 44%; melting point: 219.9-220.7℃; white powder solid. 1 H NMR (400MHz, CDCl3) δ5.71 (s, 1H, CH-12), 4.89-4.81 (d, J = 8.8Hz, 1H, Leu-CHNH), 4.58-4.50 (dd, J = 4 .7,11.7Hz,1H,CH-3),4.34-4.23(dt,J=4.5,9.0Hz,1H,Leu-CHNH),3.72-3.63(t,4H,J=5.1Hz,N(CH 2)2),2.87-2.73(m,1H,CH-1),2.52-2.40(t,J=4.9Hz,4H,N(CH2)2),2.36(s,3H,NCH3),2.04(s,1H, CH-18),2.10-1.92,1.88-1.81,1.75-1.69,1.66-1.54,1.05-0.98,0.92,0.84,0.83(22H,methylene and methine oftriterpenoid structure andleucine),1.42(s,9H,Boc-CH3),1.36(s,3H,CH3-29),1.20(s,3H,CH3-25),1.15(s,3H,CH3-26),1.11(s,3H ,CH3-27),0.96-0.93(t,J=3.3Hz,6H,Leu-CH3),0.88(s,3H,CH3-23),0.86(s,3H,CH3-24),0.80(s,3H,CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.88 (C-11), 173.96 (Leu-COO), 173.12 (C-13), 169.54 (C-30), 155.36 (Boc-COO), 128.59 (C-12), 81.60 (C-3), 79.64 (Boc-q.C), 61.66 (C-9), 55.05 (C-5), 54.54 (N(CH2)2, 2C), 52.55 (Leu-CHNH), 48.10 (C-18), 45.28 (N(CH2)2, 2C), 45.12 (NCH3), 43.88 (C-19), 43.81 (C-8), 43.28 (C-14), 42.01 (Leu-CHCH2), 38.76 (C-4), 38.14 (C-1), 37.74 (C-9), 36.93 (C-20), 33.13 (C-21), 32.73 (C-22), 31.76 (C-7), 28.43 (C-17), 28.35 (Boc-CH3, 3C), 28.12 (C-16), 26.99 (C-28), 26.71 (C-15), 26.42 (C-27), 24.85 (Leu-CH2CH), 23.52 (C-29), 23.11 (C-2), 22.97 (Leu-CH3, 2C), 18.69 (C-23 / C-24, 2C), 17.35 (C-6), 16.77 (C-26), 16.38 (C-25); IR νmax 2958, 2933, 2870, 1712, 1658, 1529, 1365, 1249, 1166, 1049, 1020, 981, 873 cm -1 ; HRMS (ESI) m / z calcd for C 46 H 76 N3O6 + (M+H) + 766.57286, found 766.57306.

[0105] Example 22

[0106]

[0107] Weigh 1.00 mmol of the compound shown in Formula 2, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Boc-L-methionine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 2 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with dichloromethane:methanol = 20:1 to 12:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 22. The physical properties and characterization data of the obtained compound are as follows: yield: 76%; melting point: 150.1-151.1℃; white powder solid. 1 H NMR (400MHz, CDCl3) δ5.66 (s, 1H, CH-12), 5.11 (d, J = 8.7Hz, 1H, Met-CHNH), 4.54 (dd, J = 11.7, 4.8Hz ,1H,CH-3),4.43-4.29(m,1H,Met-CHNH),4.24-3.89(m,4H,N(CH2)2),3.26-2.99(m,4H,N(CH2)2),2 .82(s,3H,NCH3),2.77(d,J=3.8Hz,1H,CH-1),2.56-2.51(m,2H,Met-CH2S),2.34(s,1H,CH-9),2.28 ,2.19-2.11,1.95-1.88,1.83,1.79-1.66,1.65-1.54,1.48,1.38,1.31-1.22,1.02(19H,methylene and methine oftriterpenoid structure),2.09(s,3H,Met-CH3),2.04-1.97(m,2H,Met-CH2CH),1.43(s,9H,Boc-CH3),1.34(s,3H,CH3-29),1.20(s, 3H,CH3-25),1.14(s,3H,CH3-26),1.10(s,3H,CH3-27),0.88(s,3H,CH3-23),0.86(s,3H,CH3-24),0.79(s,3H,CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.88 (C-11), 174.28 (Met-COO), 172.00 (C-13), 169.46 (C-30), 155.34 (Boc-COO), 128.58 (C-12), 82.11 (C-3), 79.91 (Boc-q.C), 61.66 (C-9), 55.00 (C-5), 53.87 (Met-CHNH), 53.25 (N(CH2)2, 2C), 47.85 (C-18), 45.28 (NCH3), 44.06 (C-8), 43.88 (N(CH2)2, 2C), 43.30 (C-14), 42.67 (C-19), 38.73 (C-4), 38.13 (C-1), 37.66 (C-10), 36.93 (C-20), 32.70 (C-21), 32.53 (C-22), 32.41 (C-7), 31.74 (C-17), 30.03 (Met-CH2CH), 28.38 (Met-CH2S), 28.33 (Boc-CH3, 3C), 28.18 (C-2), 26.91 (C-29), 26.63 (C-16), 26.33 (C-15), 23.53 (C-28), 23.10 (C-27), 18.69 (C-23 / C-24, 2C), 17.33 (C-6), 16.80 (C-26), 16.39 (C-25), 15.47 (Met-CH3); IR νmax 2974, 2931, 2791, 1712, 1658, 1525, 1452, 1390, 1365, 1290, 1166, 1049, 1001, 983, 875 cm -1 ; HRMS (ESI) m / z calcd for C 45 H 74 N3O6S + (M+H) + 784.52928, found 784.52966.

[0108] Example 23

[0109]

[0110] Weigh 1.00 mmol of the compound shown in Formula 2, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Boc-L-isoleucine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 2 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with dichloromethane:methanol = 20:1 to 12:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 23. The physical properties and characterization data of the obtained compound are as follows: yield: 76%; melting point: 214.6-216.1℃; white powder solid. 1 H NMR (400MHz, CDCl3) δ5.69(s,1H,CH-12),4.98(d,J=8.9Hz,1H,CH-3),4.54(dd,J=11.3,4.8Hz,1H,Ile-CHN H),4.24(dd,J=8.7,3.8Hz,1H,Ile-CHNH),3.68-3.63(t,J=4.8Hz,4H,N(CH2)2),2.83-2.76(dt,J=3.7,13.7 Hz,1H,CH-1),2.44-2.39(t,J=5.0Hz,4H,N(CH2)2),2.33(s,1H,CH-9),2.31(s,3H,NCH3),2.04(s,1H,CH-1 8),2.06,1.99-1.93,1.87-1.78,1.74-1.58,1.55,1.49,1.41,1.02,0.99,0.90,0.84,0.82(22H,methylene and methine of triterpenoidstructure and isoleucine),1.42(s,9H,Boc-CH3),1.34(s,3H,CH3-29),1.20(s,3H,CH3-25),1.14(s,3H,CH3-26),1.11( s,3H,CH3-27),0.96-0.91(m,6H,Ile-CH3),0.88(s,3H,CH3-23),0.87(s,3H,CH3-24),0.80(s,3H,CH3-28);13 C NMR (101 MHz, CDCl3) δ 199.88 (C-11), 173.86 (Ile-COO), 172.01 (C-13), 169.64 (C-30), 155.61 (Boc-COO), 128.58 (C-12), 81.92 (C-3), 79.60 (Boc-q.C), 61.66 (C-9), 58.50 (Ile-CHNH), 55.16 (N(CH2)2, 2C), 55.08 (C-5), 48.18 (C-18), 45.91 (NCH3), 45.29 (N(CH2)2, 2C), 43.80 (C-8), 43.29 (C-14), 38.78 (C-19), 38.04 (C-4), 37.97 (C-1), 37.76 (C-20), 36.92 (Ile-CHCH3), 33.32 (C-21), 32.74 (C-22), 31.76 (C-7), 28.45 (C-17), 28.35 (Boc-CH3, 3C), 28.09 (C-2), 27.04 (C-29), 26.74 (C-16), 26.45 (C-15), 24.73 (Ile-CH2CH3), 23.61 (C-28), 23.11 (C-27), 18.69 (C-23 / C-24, 2C), 17.37 (C-6), 16.85 (C-26), 16.37 (C-25), 15.73 (Ile-CHCH3), 11.67 (Ile-CH2CH3); IR νmax 2968, 2873, 2791, 1720, 1658, 1629, 1525, 1460, 1413, 1365, 1290, 1213, 1168, 1083, 1047, 983, 866 cm -1 ; HRMS (ESI) m / z calcd for C 46 H 76 N3O6 + (M+H) + 766.57286, found 766.57349.

[0111] Example 24

[0112]

[0113] Weigh 1.00 mmol of the compound shown in Formula 2, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Boc-L-phenylalanine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 2 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with dichloromethane:methanol = 20:1 to 12:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 24. The physical properties and characterization data of the obtained compound are as follows: yield: 75%; melting point: 132.4-135.9℃; white powder solid. 1 H NMR(400MHz, CDCl3) δ7.29(d,J=7.4Hz,2H,H-Ar),7.24(d,J=7.7Hz,1H,H-Ar),7.20-7.14(m ,2H,H-Ar),5.70(s,1H,CH-12),4.88(d,J=8.6Hz,1H,Phe-CHNH),4.66-4.48(m,2H,CH-3and Phe-CHNH),3.67(t,J=5.2Hz,4H,N(CH2)2),3.17-2.98(m,2H,Phe-CH2),2.79(d,J=13.4Hz,1H,CH-1),2.43(t,J=4.7Hz,4 H,N(CH2)2),2.33(s,3H,NCH3),2.04(s,1H,CH-18),2.28,2.14-1.96,1.82,1.69-1.41,1.01,1.25,0.88(19H,methylene and methine oftriterpenoidstructure),1.39(s,9H,Boc-CH3),1.35(s,3H,CH3-29),1.21(s,3H,CH 3-25),1.13(s,3H,CH3-26),1.11(s,3H,CH3-27),0.81(s,9H,CH3-23 / CH3-24 / CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.90 (C-11), 173.87 (Met-COO), 171.79 (C-13), 169.67 (C-30), 155.06 (Boc-COO), 136.11 (Phe-C), 129.38 (Phe-C, 3C), 128.58 (C-12), 128.54 (Phe-C), 126.96 (Phe-C), 82.16 (C-3), 79.78 (Boc-q.C), 61.66 (C-9), 55.16 (N(CH2)2, 2C), 55.05 (C-5), 54.66 (Phe-CHNH), 48.18 (C-18), 45.90 (NCH3), 45.28 (N(CH2)2, 2C), 43.85 (C-8), 43.81 (C-14), 43.29 (C-19), 38.77 (C-4), 38.44 (Phe-CH2), 38.06 (C-1), 37.76 (C-10), 36.90 (C-20), 33.31 (C-21), 32.73 (C-22), 31.77 (C-7), 28.45 (C-17), 28.32 (Boc-CH3, 3C), 28.05 (C-2), 27.05 (C-29), 26.74 (C-16), 26.45 (C-15), 23.44 (C-28), 23.11 (C-27), 18.68 (C-23 / C-24, 2C), 17.33 (C-6), 16.70 (C-26), 16.38 (C-25); IR νmax 2974, 2931, 2791, 1720, 1658, 1620, 1525, 1452, 1390, 1365, 1290, 1249, 1166, 1049, 1001, 983, 863, 766 cm -1 ; HRMS (ESI) m / z calcd for C 49 H 74 N3O6 + (M+H) + 800.55721, found 800.55771.

[0114] Example 25

[0115]

[0116] Weigh 1.00 mmol of the compound shown in Formula 2, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve them, and then weigh 1.20 mmol of Boc-L-proline into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 2 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with dichloromethane:methanol = 20:1 to 12:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 25. The physical properties and characterization data of the obtained compound are as follows: yield: 72%; melting point: 179.9-180.3℃; white powder solid. 1 H NMR (400MHz, CDCl3) δ5.69 (d, J=5.6Hz, 1H, CH-12), 4.52 (dt, J=11.7, 6.1Hz, 1H, CH-3),4.21-4.24(m,1HPro-CH),3.79-3.63(m,4H,N(CH2)2),3.48-3.42(m,2H,P ro-CH2),2.85-2.72(m,1H,CH-1),2.57(t,J=5.1Hz,4H,N(CH2)2),2.38(s,3H,NC H3),2.34(s,1H,CH-9),2.07-1.90,1.85,1.61,1.44,1.25,1.01(23H,methylene and methine of triterpenoid structure andproline),1.40(s,9H,Boc-CH3),1.34(s,3H,CH3-29),1.20(s,3H,CH3-25),1.14(s ,3H,CH3-26),1.10(s,3H,CH3-27),0.87(s,6H,CH3-23 / CH3-24),0.79(s,3H,CH3-28); 13¹³C NMR (101 MHz, CDCl₃) δ 199.79 (C-11), 173.92 (Pro-COO), 172.76 (C-13), 169.54 (C-30), 154.11 (Boc-COO), 128.53 (C-12), 81.03 (C-3), 79.86 (Boc-q.C), 79.47 (Pro-CH), 61.61 (C-9), 59.60 (C-5), 59.35 (Pro-CH₂N), 54.98 (N(CH₂)₂, 2C), 54.36 (N(CH₂)₂, 2C), 47.97 (C-18), 45.23 (NCH₃), 44.88 (C-8), 43.75 (C-14), 43.23 (C-19), 38.69 (C-4), 38.08 (C-1), 37.71 (C-10), 36.87 (C-21), 32.68 (C-20), 31.70 (C-22), 31.02 (C-7), 30.14 (C-17), 29.79 (Pro-CHCH₂), 28.39 (Boc-CH₃, 3C), 28.33 (C-16), 28.16 (C-28), 26.92 (C-15), 26.65 (C-27), 26.37 (Pro-CH₂CH₂CH₂), 23.44 (C-29), 23.06 (C-2), 18.64 (C-23 / C-24, 2C), 17.30 (C-6), 16.68 (C-26), 16.34 (C-25); IR νmax 2974, 2872, 1743, 1701, 1658, 1624, 1463, 1396, 1363, 1163, 1085, 974, 863, 771 cm -1 ; HRMS (ESI) m / z calcd for C 45 H 72 N₃O₆ + (M+H) + 750.54156, found 750.54205.

[0117] Example 26

[0118]

[0119] Weigh 1.00 mmol of the compound shown in Formula 2, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Boc-N-methyl-L-alanine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 2 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with dichloromethane:methanol = 20:1 to 12:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 26. The physical properties and characterization data of the obtained compound are as follows: yield: 68%; melting point: 126.9-127.7℃; white powder solid. 1 H NMR(400MHz, CDCl3)δ5.71-5.66(s,1H,CH-12),4.55-4.46(m,2H,CH-3 / N-Methyl-Ala-CH) ,3.66-3.57(t,J=5.1Hz,4H,N(CH2)2),2.84-2.74(br,3H,N-Methyl-Ala-NCH3),2.40-2.36 (t,J=4.9Hz,4H,N(CH2)2),2.33(s,1H,CH-9),2.28(s,3H,NCH3),2.09-2.00,1.98-1.90,1 .87-1.73,1.67-1.57,1.55-1.51,1.49,1.39-1.35,1.25-1.23,1.04-0.96(20H,methylene andmethine oftriterpenoid structure),1.44(s,9H,Boc-CH3),1.42(s,3H,N-Methyl-Ala-CHCH3),1.33(s,3H,CH3-29),1.19(s,3H ,CH3-26),1.13(s,3H,CH3-25),1.10(s,3H,CH3-27),0.85(s,6H,CH3-23 / CH3-24),0.79(s,3H,CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.90 (C-11), 173.84 (N-Methyl-Ala-COO), 172.15 (C-13), 169.68 (C-30), 155.85 (Boc-COO), 128.54 (C-12), 81.38 (C-3), 80.12 (N-Methyl-Ala-CH), 79.79 (Boc-q.C), 61.64 (C-9), 55.17 (N(CH2)2, 2C), 54.97 (C-5), 48.15 (C-18), 45.93 (NCH3), 45.27 (N(CH2)2, 2C), 43.84 (C-8), 43.78 (C-14), 43.27 (C-19), 38.72 (C-4), 38.06 (C-1), 37.74 (C-10), 36.91 (C-20), 33.26 (C-21), 32.70 (C-22), 31.74 (C-7), 31.55 (N-Methyl-Ala-N-CH3), 28.43 (C-17), 28.38 (Boc-CH3, 3C), 28.04 (C-2), 27.02 (C-29), 26.71 (C-16), 26.43 (C-15), 23.45 (C-25), 23.11 (C-27), 18.67 (C-23 / C-24, 2C), 17.35 (C-6), 16.39 (C-26), 15.49 (C-25), 14.11 (N-Methyl-Ala-CHCH3); IR νmax 2970, 2931, 2873, 1735, 1658, 1631, 1462, 1388, 1365, 1249, 1217, 1153, 1139, 1093, 1001, 985, 864, 777 cm -1 ; HRMS (ESI) m / z calcd for C 44 H 72 N3O6 + (M+H) + 738.54156, found 738.54236.

[0120] Example 27

[0121]

[0122] Weigh 1.00 mmol of the compound shown in Formula 2, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of BOC-L-valine into the above mixture and dissolve it. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 2 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (elution with dichloromethane:methanol = 20:1 to 12:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 27. The physical properties and characterization data of the obtained compound are as follows: yield: 68%; melting point: 163.2-163.9℃; white solid powder. 1 H NMR (400MHz, CDCl3) δ5.71 (s, 1H, CH-12), 5.00 (d, J = 9.2Hz, 1H, Boc-L-valine CHNH),4.56(dd,J=11.6,4.7Hz,1H,CH-3),3.65(t,J=5.0Hz,4H,NH(CH2)2),2.82(dt,J=13.6,3.6Hz,1H,Boc-L-va lineCHNH),2.41(t,J=4.9Hz,4H,NH(CH2)2),2.35(s,1H,CH-9),2.31(s,3H,NCH3),2.30-2.24(m,1H,Boc-L-valine CHCH3),2.19(dt,J=7.2,3.7Hz,1H,CH-18),2.05,1.98,1.91-1.76,1.76-1.70,1.70-1.59,1.54,1.39,1.39,1.26,0.82(19H,methylene and methineoftriterpenoid structure),1.45(s,9H,Boc-CH3),1.36(s,3H,CH-25),1.22(s,3H,CH-26),1.16(s,3H,CH-27),1.12 (s,3H,CH-28),0.99(d,J=6.9Hz,6H,CH-23 / CH-24),0.89(dd,J=6.1,2.9Hz,9H,CH-29 / Boc-L-valine CHCH3); 1313C NMR (101 MHz, CDCl3) δ 199.90 (C-11), 173.88 (L-valine-COO), 172.10 (C-13), 169.65 (C-30), 155.74 (Boc-COO), 128.60 (C-12), 81.89 (C-3), 79.62 (Boc-q.C), 61.66 (C-9), 58.90 (NHCH), 55.17 (C-5), 55.07 (NCH3), 48.18 (C-18), 45.92 (N(CH2)2, 2C), 45.29 (N(CH2)2, 2C), 43.80 (C-8), 43.29 (C-14), 38.77 (C-19), 38.07 (C-4), 37.76 (C-1), 36.93 (C-10), 33.33 (C-20), 32.74 (C-21), 31.77 (C-7 / C-22, 2C), 31.18 (Boc-L-valine CH(CH3)2), 28.45 (C-17), 28.35 (Boc-CH3, 3C), 28.12 (C-2), 27.05 (C-28), 26.74 (C-29), 26.45 (C-16), 23.63 (C-15), 23.12 (C-27), 19.33 (Boc-L-valine CH(CH3)2, 2C), 18.69 (C-23), 17.37 (C-24), 17.21 (C-6), 16.83 (C-26), 16.37 (C-25); IR vmax 2972, 2873, 2358, 1729, 1647, 1612, 1500, 1161, 987, 879 cm -1 ; HRMS (ESI) m / z calcd for C 45 H 73 N3O6Na + (M+Na) + 774.53916, found 774.53857.

[0123] Example 28

[0124]

[0125] Weigh 1.00 mmol of the compound shown in Formula 2, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of BOC-L-aspartic acid-4-benzyl ester and add it to the above mixture to dissolve. The mixture was stirred at room temperature, and the reaction was monitored at regular intervals using thin-layer chromatography. Stirring was stopped when a large amount of the target product was generated and the amount of the compound shown in Formula 2 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, followed by two extractions with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then purified by silica gel column chromatography (dichloromethane:methanol = 20:1 to 12:1 elution of petroleum ether:ethyl acetate:methanol = 4:1:0.5). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 28. The physical properties and characterization data of the obtained compound are as follows: yield: 47%; melting point: 118.3-121.1℃, yellow solid powder. 1 H NMR (400MHz, CDCl3) δ7.40-7.29(m,5H,H-Ar),5.71(s,1H,CH-12),5.52(d,J=8.9Hz,1H,NHCH),5.11( s,2H,Ar-CH2),4.57-4.53(m,1H,CH-3),3.65(d,J=5.1Hz,4H,N(CH2)2),2.92-2.74(m,2H),2.41(t,J= 5.0Hz,4H,N(CH2)2),2.32(s,3H,NCH3),2.06(td,J=9.2,3.4Hz,2H,CH2COO),1.96(t,J=3.3Hz,1H,CH- 9),1.83(d,J=4.6Hz,1H,CH-18),1.67,1.57-1.50,1.40,1.26,1.08-0.95,0.88,0.83(19H,methylene and methineof triterpenoid structure),1.66-1.57(s,3H,CH-25),1.44(s,9H,Boc-CH3),1.36(s,3H,CH-26),1.22(s ,3H,CH-27),1.15(s,3H,CH-28),1.12(s,3H,CH-29),0.81(d,J=2.8Hz,6H,CH-23 / CH-24); 13¹³C NMR (101 MHz, CDCl₃) δ 199.90 (C-11), 173.88 (COO, 2C), 170.98 (CONH), 170.78 (C-13), 169.68 (C-30), 155.45 (Boc-COO), 135.36 (Ar-C), 128.61 (Ar-C), 128.59 (Ar-C), 128.47 (Ar-C), 128.42 (Ar-C / C-12, 2C), 128.37 (Ar-C), 82.49 (C-3), 79.97 (Boc-q.C), 66.82 (Ar-CH₂), 61.65 (C-9), 55.18 (NCH₃), 55.05 (NHCH), 50.20 (C-5), 48.18 (C-18), 45.94, 45.29 (N(CH₂)₂, 2C), 43.85 (N(CH₂)₂, 2C), 43.81 (C-8), 43.30 (C-14), 38.74 (C-19), 38.07 (C-4), 37.77 (C-1), 36.91 (C-10), 36.74 (CH₂), 33.33 (C-20), 32.73 (C-21), 31.77 (C-22 / C-7, 2C), 28.45 (C-17), 28.33 (Boc-CH₃, 3C), 27.89 (C-2), 27.05 (C-29), 26.75 (C-28), 26.46 (C-16), 23.38 (C-27), 23.13 (C-23), 18.69 (C-24), 17.33 (C-6), 16.70 (C-26), 16.37 (C-25); IR vmax 2974, 2873, 2358, 2341, 1743, 1622, 1516, 1454, 1423, 1364, 1290, 1165, 1139, 1001, 748, 696 cm -1 ; HRMS (ESI) m / z calcd for C 51 H 75 N₃O₈Na + (M+Na) + 880.54464, found 880.54456.

[0126] Example 29

[0127]

[0128] Weigh 1.00 mmol of the compound shown in Formula 2, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of BOC-tryptophan and add it to the above mixture to dissolve. The mixture was stirred at room temperature, and the reaction was monitored by thin-layer chromatography at regular intervals. Stirring was stopped when a large amount of the target product was generated and the compound shown in Formula 2 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane and then twice with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was then separated and purified by silica gel column chromatography (elution with dichloromethane:methanol = 20:1 to 12:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 29. The physical properties and characterization data of the obtained compound are as follows: Yield: 63%, white solid powder; 1 H NMR (400MHz, CDCl3) δ8.42-8.34(m,1H,C=C-NH),7.60(d,J=7.9Hz,1H,H-Ar),7.33(d,J=8.0 Hz,1H,H-Ar),7.18-7.09(m,2H,H-Ar),6.99(d,J=2.3Hz,1H,CH=C),5.71(s,1H,CH-12),5.1 2(d,J=8.2Hz,1H,CHNH),4.65(q,J=6.4Hz,1H,CH3),4.12(q,J=7.1Hz,1H,CHNH),3.63(s,4H ,N(CH2)2),2.84-2.68,1.97,1.63,1.52,1.39,1.26,1.17-1.13,0.77,0.73(19H,methylene and methhine of triterpenoidstructure),2.40(s,4H,N(CH2)2),2.30(s,3H,CH3),2.08(s,1H,CH-18),2.04(s,3H),1.43(s,9H,Boc-CH3),1.34(s,3H,CH-26),1.26(t,J =7.1Hz,2H),1.21(s,3H,CH-27),1.17-1.13(m,1H),1.11(d,J=2.3Hz,6H,CH -28 / CH-29),1.01(d,J=13.4Hz,2H),0.81(s,3H,CH-23),0.78(s,3H,CH-24); 13¹³C NMR (101 MHz, CDCl₃) δ 200.08 (C-11), 173.93 (COO), 172.15 (C-13), 169.89 (C-30), 155.29 (Boc-COO), 136.19 (NH-C=C), 128.57 (C-12), 127.88 (Ar-C), 122.81 (Ar-C), 122.06 (Ar-C), 119.45 (Ar-C), 119.04 (Ar-C), 111.14 (Ar-C), 110.33 (CH=C), 81.94 (C-3), 61.68 (Boc-q.C), 60.41 (C-9), 55.11 (C-5), 55.01 (NCH₃), 54.41 (NHCH), 48.20 (C-18), 45.85 (N(CH₂)₂, 2C), 45.30 (N(CH₂)₂, 2C), 43.82 (C-8), 43.31 (C-14), 38.74 (C-19), 38.03 (C-4), 37.77 (C-1), 36.90 (C-10), 33.30 (C-20), 32.71 (C-21), 31.77 (C-22 / C-7, 2C), 28.46 (C-17), 28.36 (Boc-CH₃, 3C), 28.13 (C-2), 27.92 (C-29), 27.05 (C-28), 26.73 (C-16), 26.44 (C-15), 23.24 (C-27), 23.12 (C-23), 18.68 (C-24), 17.33 (C-6), 16.58 (C-25); IR vmax 3264, 2873, 1712, 1492, 1458, 1215, 1049, 860, 740, 673 cm -1 .

[0129] Example 30

[0130]

[0131] Weigh out 1.00 mmol of the compound shown in Formula 2, 1.20 mmol of EDCI, and DMAP. 0.20 mmol was placed in a 20 mL reaction tube and dissolved in 10 mL of dry dichloromethane. Then, 1.20 mmol of (N-(tert-butyloxycarbonyl)-N-methylglycine) was weighed and added to the above mixture to dissolve it. The mixture was stirred at room temperature, and the reaction was monitored by thin-layer chromatography at regular intervals. Stirring was stopped when a large amount of the target product was generated and the compound shown in Formula 2 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane, and then twice with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane:methanol = 20:1 to 12:1 elution). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 30. The physical properties and characterization data of the obtained compound are as follows: Yield: 63%, white solid powder; 1 H NMR (400MHz, CDCl3) δ5.71 (d, J=3.0Hz, 1H, CH-12), 4.57 (ddd, J=11.1, 8.5, 5. 0Hz,1H,CH-3),4.01-3.85(m,2H,NCH2),3.68(t,J=4.9Hz,4H,N(CH2)2),2.91( d,J=2.8Hz,3H,NCH3),2.47(t,J=4.9Hz,4H,N(CH2)2),2.34(s,3H,NCH3),2.04 (s,1H,CH-18),1.96,1.84,1.79-1.49,1.47,1.38,1.26,0.99(19H,methylene and methine oftriterpenoid structure),1.43(s,9H,Boc-CH3),1.36(s,3H CH3-29),1.22(s,3H,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-27),0.88(dd,J=6.2,2.7Hz,6H,CH3-23 / CH3-24),0.82(s,3H,CH3-28); 1313C NMR (101 MHz, CDCl3) δ 199.90 (C-11), 173.91 (C-13), 169.77 (COO), 169.68 (C-30), 155.94 (Boc-COO), 128.57 (C-12), 81.59 (C-3), 80.10 (Boc-q.C), 61.65 (C-9), 55.02 (C-5), 54.85 (N(CH2)2, 2C), 51.29 (NCH3), 50.72 (NCH2), 48.13 (C-18), 45.54 (NCH3), 45.28 (N(CH2)2, 2C), 43.88 (C-8), 43.79 (C-14), 43.28 (C-19), 38.76 (C-4), 38.07 (C-1), 37.74 (C-10), 36.91 (C-20), 33.19 (C-21), 32.71 (C-22), 31.75 (C-7), 28.43 (C-17), 28.34 (Boc-CH3, 3C), 28.29 (C-2), 28.10 (C-29), 27.00 (C-16), 26.71 (C-15), 26.42 (C-28), 23.58 (C-27), 23.10 (C-23), 18.67 (C-24), 17.36 (C-6), 16.66 (C-26), 16.40 (C-25); IR vmax 3435, 2976, 2875, 1707, 1570, 1429, 1095, 923, 833 cm -1 ; HRMS (ESI) m / z calcd for C 43 H 69 N3O9Na + (M+Na) + 830.49260, found 830.49274.

[0132] Example 31

[0133]

[0134] Weigh 1.00 mmol of the compound shown in Formula 2, 1.20 mmol of EDCI and 0.20 mmol of DMAP into a 20 mL reaction tube, add 10 mL of dry dichloromethane to dissolve, and then weigh 1.20 mmol of Boc-L-pyroglutamic acid and add it to the above mixture to dissolve. The mixture was stirred at room temperature, and the reaction was monitored by thin-layer chromatography at regular intervals. Stirring was stopped when a large amount of the target product was generated and the compound shown in Formula 2 no longer decreased. The mixture was then poured into 15 mL of ice water to terminate the reaction. The mixture was then transferred to a separatory funnel and extracted twice with 15 mL of dichloromethane and then twice with saturated sodium chloride aqueous solution to remove water-soluble impurities. The mixture was then dried with anhydrous sodium sulfate to remove excess water, filtered, and concentrated under reduced pressure to obtain the target crude product. The crude product was separated and purified by silica gel column chromatography (elution with dichloromethane:methanol = 20:1 to 12:1). After purification, the mixture was concentrated under reduced pressure to obtain the compound shown in Formula 31. The physical properties and characterization data of the obtained compound are as follows: Yield: 50%, white solid powder; 1 H NMR(400MHz,CDCl3)δ5.72(s,1H,CH-12),4.68-4.45(m,2H,CH-3 / NCH),3.68(dd,J=6.1,3.3Hz,4H,N(CH2)2),2.48-2.42(m,4H,N(CH2)2),2 .34(s,4H,COCH2CH2),2.11-2.04(m,3H,NCH3),2.04-1.89,1.70-1.61,1.61-1.52,1.48,1.47-1.38,1.03,0.97-0.89,0.80(19H,methylene and methhine of triterpenoidstructure),1.89-1.70(m,2H,CH-18 / CH-9),1.50(s,9H,Boc-CH3),1.36(s,3H,CH3-29),1.22(s,3H ,CH3-25),1.16(s,3H,CH3-26),1.12(s,3H,CH3-27),0.88(d,J=5.7Hz,6H,CH3-23 / CH3-24),0.82(s,3H,CH3-28); 13C NMR(101MHz,CDCl3)δ199.84(C-11),173.89(CON),173.29(C-13),170.96(CO O),169.78(C-30),149.41(Boc-COO),128.54(C-12),83.62(C-3),82.18(Boc -qC),61.61(C-9),59.32(N(CH2)2,2C),55.02(C-5),48.10(C-18),45.73(NC H3),45.25(N(CH2)2,2C),43.91(CH2COO),43.80(C-8 / C-14,2C),43.28(C-19) ,38.67(C-4),38.12(C-1),37.74(C-10),36.89(C-20),33.14(C-21),32.67( C-22),31.74(C-7),31.19(NCOCH2),28.43(C-17 / Boc-CH3,4C),28.16(C-2),2 7.94(C-29),27.00(C-16),26.70(C-15),26.42(C-28),23.52(C-27),23.09( C-23),21.97(CH2),18.66(C-24),17.32(C-6),16.72(C-26),16.37(C-25); IR vmax 3157,2791,1764,1639,1570,1369,1251,1101,921,881cm -1 HRMS(ESI)m / z calcd for C 45 H 69 N3O7Na + (M+Na) + 786.50277, found 786.50232.

[0135] Effect Experiment

[0136] Cytotoxicity test of the compounds shown in Formula 1-31 against the MDA-MB-231 cell line

[0137] 1.1 Experimental Materials

[0138] 1.1.1 Cell line: Human breast cancer cells MDA-MB-231 were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences and cultured in high glucose DMEM medium (Gibco), 10% FBS serum (Gibco), 5% CO2, and a constant temperature incubator at 37°C. The cells were passaged and related experimental procedures were performed when the confluence reached about 70-80%.

[0139] 1.1.2 Materials and reagents: fetal bovine serum (BI, USA), DMEM high glucose medium (Gibco, USA), penicillin-streptomycin mixture (Solepro, Shanghai), PBS buffer solution (Gibco, USA), isopropanol (analytical grade), dimethyl sulfoxide (DMSO) (Solepro, Shanghai), MTT powder (Solepro, Shanghai).

[0140] 1.2 Experimental Methods:

[0141] 1.2.1 Assay on the anti-proliferative activity of 18β-GA derivatives against triple-negative breast cancer

[0142] Sample solution: Accurately weigh the sample using a balance of 0.01%, then dissolve it in biological grade DMSO to a suitable concentration. Store the stock solution at -20℃. Before use, dilute it to the required concentration with the required incomplete culture medium.

[0143] Incomplete culture medium: 99% high glucose DMEM medium, add 1% penicillin (100U / mL)-streptomycin (100μg / mL) (double antibiotics), and mix well for later use.

[0144] MDA-MB-231 cell complete culture medium: 90% high glucose DMEM medium, with 1% penicillin antibiotics and 10% fetal bovine serum added, and mixed thoroughly for later use.

[0145] Preparation of MTT solution: Accurately weigh 0.10 g of MTT powder using a 0.01% balance, dissolve it in 20 mL of phosphate buffered saline (PBS) solution to a final concentration of 5 mg / mL, filter through a 0.22 μm microporous membrane, and store at 4°C protected from light. During preparation and storage, wrap the container with aluminum foil.

[0146] Take the cryopreserved MDA-MB-231 cells and quickly transfer them to a 37°C water bath, shaking the cryovial continuously to rapidly thaw the cells (within 1-2 minutes). After the culture medium has thawed, mix the cell suspension well, transfer it to a 15mL centrifuge tube, centrifuge at 1000rpm for 5 minutes, and remove the cryopreservation solution. Then, resuspend the cells in complete cell culture medium, transfer them to a culture flask, mix thoroughly, observe the cell state, and record the results. Incubate at 37°C in a 5% CO2 incubator.

[0147] After 24 hours of culture, observe cell density and growth status. When the density is 70-90% and the cells are in good condition, they can be passaged (1:2). Take cells in the logarithmic growth phase, discard the culture medium, add PBS buffer along the cell wall, and wash thoroughly to ensure the cells adhered to the bottom. Discard the PBS buffer. Then add trypsin digestion solution to digest the cells. After the cells have fully contacted the trypsin, discard the trypsin and let stand for 1-3 minutes. When cells are observed to slip off visually, add complete culture medium to stop the digestion. Then, transfer 0.5 mL to a new culture flask, add 3-4 mL of complete culture medium to the remaining 0.5 mL of cell suspension, mix well, and continue culturing.

[0148] (1)Planning

[0149] Cells in the logarithmic growth phase were passaged until trypsin digestion was complete. 1 mL of fresh complete culture medium was added, and the cells were thoroughly resuspended by pipetting. 0.5 mL of this resuspended cell suspension was transferred to a centrifuge tube. 3-4 mL of complete culture medium was added to the remaining 0.5 mL of cell suspension, and the tube was incubated at 37°C with 5% CO2. The cell suspension in the centrifuge tube was then diluted with an appropriate amount of complete culture medium. 10 μL of the cell suspension was diluted with 10 μL of PBS buffer, and after thorough mixing, 10 μL was transferred to a hemocytometer. The cells were observed and counted under an upright microscope. Cells were counted at a rate of 1 × 10⁻⁶. 4 Cell density was calculated per well. Cells were seeded into 96-well plates, with 100 μL of cell suspension added to each well. PBS buffer was added around the perimeter of the 96-well plate. The plates were then incubated at 37°C with 5% CO2 for 24 hours before drug administration.

[0150] (2) Administration

[0151] 24 hours after cell seeding, the 96-well plates were removed, and the remaining complete culture medium was aspirated. Five concentration gradients were selected for each drug, with four replicates per concentration. Control groups (no compound), blank groups (no cells), and positive control groups (cells and positive control drug) were set up. 100 μL of the prepared compound at different concentrations was added to each well of the 96-well plate. After drug administration, the plates were incubated at 37°C with 5% CO2 for 48 hours, after which the MTT assay was performed.

[0152] (3) MTT test

[0153] After culturing cells for 48 hours following drug administration, the incomplete culture medium containing the compound was aspirated in the dark. 110 μL of pre-prepared MTT mixture (MTT solution: incomplete culture medium = 1:10) was added to each well, and the cells were incubated for another 4 hours. The culture medium in the wells was then discarded, and 200 μL of dimethyl sulfoxide was added to each well. The cells were then incubated on a microplate shaker for 10-15 minutes. The absorbance at 490 nm was then measured using a microplate reader, and the data was saved. The inhibition rate % was calculated using the formula: Inhibition rate % = [1 - (OD200%) / ( ... 给药组 -OD 空白组 ) / (OD 对照组 -OD 空白组 The corresponding inhibition rate was calculated by multiplying the value by 100%, and the IC50 of the drug was simulated using the software GraphPadPrism 8.0. 50 value.

[0154] 1.3 Experimental Results

[0155] Table 1 shows the IC50 of the compounds in Formula 1-31 against MDA-MB-231 breast cancer cells. 50 value

[0156] 1 7.02±1.27 18 13.60±2.90 2 7.43±0.86 19 6.81±0.28 3 5.04±0.05 20 0.75±0.05 4 3.26±0.82 21 1.16±0.31 5 26.68±1.30 22 5.84±3.55 6 3.92±2.61 23 2.84±0.85 7 15.23±2.90 24 2.56±1.36 8 2.17±1.30 25 1.73±0.22 9 13.94±5.99 26 1.57±0.28 10 ＞50 27 ＞50 11 1.85±1.06 28 10.70±0.58 12 ＞50 29 4.51±0.17 13 1.51±0.97 30 6.81±0.28 14 0.37±0.01 31 5.85±0.09 15 13.51 Doxorubicin 1.76±0.47 16 ＞50 18β-GA 45.26±0.75 17 ＞50 - -

[0157] The cytotoxic effects of 31 GA derivatives, the positive control drug doxorubicin, and the lead compound 18β-GA on the human triple-negative breast cancer cell line MDA-MB-231 were examined using the MTT assay. Most of the obtained derivatives showed stronger antiproliferative activity than the parent compounds. Compound 14 exhibited stronger antiproliferative activity against triple-negative breast cancer cells than doxorubicin, and was 122.3 times more potent than the parent compound 18β-GA, with an IC50 value of [missing value]. 50 The value was 0.37±0.01 μM. This invention obtained candidate compounds with high efficacy against triple-negative breast cancer and good drug-like properties through scientific structural modification of 18β-GA. Among them, the superior compound 14 exhibits stronger anti-triple-negative breast cancer cell proliferation activity than various 18β-GA derivatives reported to date.

[0158] The description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A type of 18 β -Glycyrrhetinic acid derivative, characterized in that... Having the following formulas: Formula 1, Formula 3, Formula I, or Formula II The structure shown is as follows: 、 Formula 1 、 Formula 3 ； Formula I or ; Formula II In Formula I, R1 is selected from 3-phenylpropionyl, Fmoc-L-methionine acyl, Boc-glycine acyl, Boc-L-leucine acyl, Boc-L-isoleucine acyl, Boc-L-proline acyl, Boc-N-methyl-L-alanine acyl, Boc-L-valine acyl, N-(tert-butoxycarbonyl)-N-methylglycine acyl or Boc-L-pyroglutamic acid acyl; In Formula II, R2 is selected from Boc-glycine acyl, Boc-L-leucine acyl, Boc-L-methionine acyl, Boc-L-isoleucine acyl, Boc-L-phenylalanine acyl, Boc-L-proline acyl, Boc-N-methyl-L-alanine acyl, 4-benzyl ester-Boc-L-aspartic acyl, Boc-L-tryptophan acyl, N-(tert-butoxycarbonyl)-N-methylglycine acyl, or Boc-L-pyroglutamic acid acyl.

2. The 18-inch steel according to claim 1 β The method for preparing glycyrrhetinic acid derivatives is characterized by, The preparation method of the compound shown in Formula 1 or Formula 3 includes the following steps: (1.1) Weigh 18 β - GA, EDCI and HOBT were placed in a reaction vessel and dissolved in anhydrous dichloromethane. Then TEA was added dropwise, and the mixture was stirred at room temperature. A polyamine compound was then added, and the mixture was stirred at room temperature to carry out the reaction. The reaction system was monitored by thin-layer chromatography. After the reaction was completed, stirring was stopped to obtain the reactants. The polyamine compound was piperazine-1-carboxylic acid methyl ester or N,N-dimethyldipropyltriamine. (1.2) Pour the reactants into ice water, transfer the mixture to a separatory funnel, extract with dichloromethane, collect the organic phase, extract with saturated sodium chloride aqueous solution, collect the organic phase, dry with anhydrous sodium sulfate, filter, concentrate under reduced pressure to obtain the target crude product, separate and purify the target crude product by silica gel column chromatography, concentrate under reduced pressure to obtain the compounds shown in Formula 1 and Formula 3.

3. The 18-inch steel according to claim 2 β The method for preparing glycyrrhetinic acid derivatives is characterized by, In step (1.1), 18 β The molar ratio of GA, EDCI, HOBT, TEA and polyamine compounds is 0.43:0.86:0.43:2.15:2.

15.

4. The 18-inch steel according to claim 1 β The method for preparing glycyrrhetinic acid derivatives is characterized by, The preparation method of the compound shown in Formula I includes the following steps: (3.1) Weigh the compound shown in Formula 1, EDCI and DMAP into a reaction vessel, add anhydrous dichloromethane to dissolve them, and then weigh 3-phenylpropionic acid, Fmoc-L-methionine, Boc-glycine, Boc-L-leucine, Boc-L-isoleucine, Boc-L-proline, Boc-N-methyl-L-alanine, Boc-L-valine, Boc-L-tryptophan, N-(tert-butoxycarbonyl)-N-methylglycine or Boc-L-pyroglutamic acid and add them to the above mixture to dissolve them. Continue to stir the mixture at room temperature to carry out the reaction. Monitor the reaction system by thin-layer chromatography. Stop stirring after the reaction is completed to obtain the reactants. (3.2) Pour the reactants into ice water, transfer the mixture to a separatory funnel, extract with dichloromethane, collect the organic phase, extract with saturated sodium chloride aqueous solution, collect the organic phase, dry with anhydrous sodium sulfate, filter, concentrate under reduced pressure to obtain the target crude product, separate and purify the target crude product by silica gel column chromatography, concentrate under reduced pressure to obtain the compound shown in Formula I.

5. The 18-inch steel according to claim 1 β The method for preparing glycyrrhetinic acid derivatives is characterized by, The preparation method of the compound shown in Formula II includes the following steps: (4.1) Weigh the compound shown in Formula 2, EDCI and DMAP and place them in a reaction vessel. Add anhydrous dichloromethane to dissolve them. Then weigh Boc-glycine, Boc-L-leucine, Boc-L-methionine, Boc-L-isoleucine, Boc-L-phenylalanine, Boc-L-proline, Boc-N-methyl-L-alanine, 4-benzyl ester-Boc-L-aspartic acid acyl, Boc-L-tryptophan, N-(tert-butoxycarbonyl)-N-methylglycine or Boc-L-pyroglutamic acid and add them to the above mixture to dissolve them. Continue to stir the mixture at room temperature to carry out the reaction. Monitor the reaction system by thin-layer chromatography. Stop stirring after the reaction is completed to obtain the reactants. ； Formula 2 (4.2) Pour the reactants into ice water, transfer the mixture to a separatory funnel, extract with dichloromethane, collect the organic phase, extract with saturated sodium chloride aqueous solution, collect the organic phase, dry with anhydrous sodium sulfate, filter, concentrate under reduced pressure to obtain the target crude product, separate and purify the target crude product by silica gel column chromatography, concentrate under reduced pressure to obtain the compound shown in formula II.

6. The 18 as claimed in claim 1 β - Application of glycyrrhetinic acid derivatives in the preparation of drugs for treating triple-negative breast cancer.

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