CN108350023B - Compound with anticancer effect and preparation method and application thereof - Google Patents

Abstract

The invention provides a compound with a structure (I), a pharmaceutically acceptable salt, a stereoisomer, a solvate, a polymorph or a prodrug thereof, a preparation method thereof, and application of a pharmaceutical composition containing the compound in preparation of an anticancer drug.

Description

Compound with anticancer effect and preparation method and application thereof

Technical Field

The invention relates to a compound and a preparation method and application thereof, in particular to a compound with selective anticancer activity and a preparation method and application thereof.

Background

The current clinical tragedies of 'death and death of cancer' often appear in the treatment of cancer, especially in various malignant canceration such as liver cancer, colon cancer, cervical cancer and the like, no specific medicine is available at present, the reason for this is that chemotherapy is one of the main modes of cancer at present, and first-line chemotherapeutic drugs often cause serious toxic and side effects to normal organisms while killing cancer cells, for example: nephrotoxicity, hepatotoxicity, neurotoxicity, and the like. The aim of obtaining the anti-Cancer medicine with high efficiency, low toxicity and strong selectivity is achieved by cumin of pharmaceutical workers, and the compound with strong cytotoxicity selectivity discovered by recent research at home and abroad has good druggability and accords with the development trend of modern precise medicine (Cancer Cell, 2015, 28, 240-252; int.J. mol.Sci.2015, 16(7), 16401-16413).

The compound is synthesized by chemical synthesis by taking cholic acid components with biological activities of resisting tumors and liver diseases, which mainly comprise Cholic Acid (CA), deoxycholic acid (DCA), ursodeoxycholic acid (UCA), hyodeoxycholic acid (HCA), chenodeoxycholic acid (CDCA), lithocholic acid (LCA) and a natural medicine ligustrazine (TMP) as starting raw materials. The activity evaluation of the compounds mainly surrounds the aspects of anti-tumor (especially liver cancer, intestinal cancer and the like), and the cytotoxic activity of the analogues on 4 cancer cell lines (HepG-2, HT-29, Hela, MCF-7), 9 primary liver cancer cell lines (HCC3787C1, HCC4172A2, HCC4477B2, HCC4484B1, HCC5850A1, HCC5864D1, HCC5953B1, HCC633A5 and HCC061A2) and non-cancer cell lines (NGF-induced PC12 cells, kidney epithelial cells MDCK and mouse embryo fibroblast 3T3) is tested respectively.

Disclosure of Invention

The first object of the present invention is to provide a compound having the structure of formula 1 and a method for preparing the same.

The second purpose of the invention is to provide the application of the compound in the formula 1 in preparing anti-cancer drugs.

The third purpose of the invention is to provide a medicament box with anticancer effect.

The purpose of the invention is realized by the following technical scheme:

a compound having the structure of formula 1, a pharmaceutically acceptable salt, stereoisomer, isotopic label, solvate, polymorph or prodrug thereof:

wherein R is₁-R₄Are independently-OH, -H or

Any one of the substituents;

r5 is-COOH,

Any one of the substituents;

and R is₁-R₅At least one of them

And (4) a substituent.

The present application further provides a pharmaceutical composition comprising a compound described herein above, a pharmaceutically acceptable salt, stereoisomer, isotopic label, solvate, polymorph or prodrug thereof, and a pharmaceutically acceptable carrier. The pharmaceutical composition includes, but is not limited to, oral dosage forms, parenteral dosage forms, topical dosage forms, and rectal dosage forms. In some embodiments, the pharmaceutical composition may be a tablet, capsule, pill, powder, sustained release formulation, solution or suspension for oral administration, a sterile solution, suspension or emulsion for parenteral injection, an ointment or cream for topical application, or a suppository for rectal administration.

The preparation method of the compound comprises the following steps:

scheme 1: dissolving cholic acid components in an organic solvent, and reacting with 2-bromomethyl-3, 5, 6-trimethylpyrazine (an intermediate compound 1) under an alkaline condition to obtain a compound I;

the compound I further reacts with ligustrazine acid (an intermediate compound 2) under the action of a catalyst and a condensing agent to obtain a compound II.

The reaction route is as follows:

reaction conditions and reagents: (a) benzene, reflux, 10 h; (b) CCl₄，NBS，hv，reflux， 12h；(c)DMF，K₂CO₃，N₂，85℃，1.5h；(d)H₂O，KMnO₄，37℃，6h；(e)dry CH₂Cl₂，EDCI，DMAP，12h。

Wherein, R in the compound I₁-R₄Are each-OH or-H, and R₁、R₂、R₄At least one-OH group;

in compound II R₃is-OH or-H, R₁、R₂、R₄Are independently-OH, -H or

And R is₁、R₂、R₄At least one of them

The specific structure of each compound is shown in Table 1.

TABLE 1 Compound I and II structural formulas

Scheme 2: dissolving cholic acid components in an organic solvent, and reacting with benzyl bromide under an alkaline condition to obtain an intermediate compound 3; the intermediate compound 3 further reacts with ligustrazine acid (intermediate compound 2) under the action of a catalyst and a condensing agent to obtain a compound III; the compound III is further reduced by hydrogen to obtain the compound IV of the invention.

The reaction route is as follows:

a，THF，Reflux，K₂CO₃；b，CH₂Cl₂，EDCI/DMAP，c，THF，Pd/C，Pd(OH)₂/C

wherein, R in the compounds III and IV₃is-OH or-H, R₁、R₂、R₄Are independently-OH, -H or

And R is₁、R₂、R₄At least one of them

The specific structure of each compound is shown in Table 2.

TABLE 2 Compound III and IV structural formulas

Further, the above reaction is carried out at-20 ℃ to 250 ℃; the organic solvent is ether, alcohol, alkane, aromatic hydrocarbon, ketone, alkyl halide, amide, nitrile, ester or their mixture in various proportion containing 1-20 carbon atoms; the alkali is triethylamine or potassium carbonate; the catalyst is 1-Hydroxybenzotriazole (HOBT); the condensing agent is any one or more of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI), 1, 3-Dicyclohexylcarbodiimide (DCC) or 4-Dimethylaminopyridine (DMAP).

Further, in the preparation method, the molar ratio of the cholic acid component to the 2-bromomethyl-3, 5, 6-trimethylpyrazine (intermediate compound 1) is 1: 0.1-1: 10; the molar ratio of the cholic acid component to the alkali is 1: 0.1-1: 10; the mol ratio of the compound II to the ligustrazine acid (the intermediate compound 2) is 1: 0.1-1: 10; the molar ratio of the compound II to the condensing agent is 1: 0.1-1: 10; the molar ratio of the catalyst to the catalyst is 1: 0.1-1: 10.

The invention also provides application of the compound shown in the formula 1, pharmaceutically acceptable salt, stereoisomer, isotopic marker, solvate, polymorph or prodrug thereof in preparing an anticancer drug.

Further, the cancer is liver cancer, colon cancer, cervical cancer, breast cancer.

The present invention also provides a method of treating cancer, comprising administering to a subject in need thereof an effective amount of a compound of formula 1, or a pharmaceutically acceptable salt, stereoisomer, isotopic label, solvate, polymorph or prodrug thereof. The subject may be a mammal, such as a human.

Further, the effective amount of the compound of formula 1 or a pharmaceutically acceptable salt, stereoisomer, isotopic label, solvate, polymorph or prodrug thereof is 1-10 mg/kg-day.

Further, the cancer is liver cancer, colon cancer, cervical cancer, breast cancer.

In some embodiments, the pharmaceutical composition and the at least one therapeutic agent are combined into a combination product, such as a kit, in separate dosage forms. The combination product comprises 1-10 mg/kg-day of effective dose of therapeutically effective dose of compound shown in formula 1 or pharmaceutically acceptable salt, stereoisomer, isotope label, solvate, polymorph or prodrug thereof.

Further, the anticancer drug is one or more of cyclophosphamide, 5-fluorouracil, paclitaxel, adriamycin, etoposide, irinotecan, oxaliplatin, cisplatin or jiaojian.

The term "pharmaceutically acceptable" as used herein refers to molecular entities and compositions that are physiologically tolerable and do not typically produce allergic or similar untoward reactions when used in humans. The "therapeutically effective amount" refers to an amount of drug administered sufficient to cause an improvement in a clinically significant condition/symptom in the host. The term "solvate" refers to a combination of a compound of the present application and a solvent molecule formed by solvation. The "polymorphs" refer to the compounds of the present application in different crystal lattice forms. The term "isotopic label" refers to a compound of the present application which is labeled with an isotope. The term "pharmaceutically acceptable prodrug" refers to any pharmaceutically acceptable salt, ester, salt of an ester, or other derivative of a compound of the present application that, upon administration to a recipient, is capable of providing, directly or indirectly, a compound of the present application or a pharmaceutically active metabolite or residue thereof. The term "stereoisomer" refers to an isomer resulting from the difference in the arrangement of atoms in a molecule.

The compound has an inhibiting effect on various cancer cells, the tumor inhibition rate of 30mg/kg by intraperitoneal injection on H22 tumor-bearing mice reaches 60.16 percent, and the tumor inhibition rate is equivalent to that of a positive medicament cyclophosphamide; has little toxicity to normal cells such as nerve cells, kidney cells, mouse embryo fibroblasts and the like, and normal mice are injected with LD in the abdominal cavity₅₀More than 900mg/kg, indicating that the medicine has high safetyIt can be used for preventing and treating cancers such as hepatocarcinoma, intestinal cancer, and cervical cancer.

Experimental example 1 AlamarBlue method for observing the effect of the composition of the present invention on the proliferation of cancer cells and non-cancer cell lines

1. Apparatus and materials

Thermo 311 type CO₂An incubator; a Haier biosafety cabinet; molecular Devices microplate reader; hunan instrumental L530 type desk type low-speed centrifuge; olympus IX51 inverted fluorescence microscope, RPMI-1640 medium, fetal bovine serum, 0.25% trypsin solution, phosphate buffer (seimer heisha shanghai ltd); sigma dimethyl sulfoxide (DMSO), resazurin;

human breast cancer cell line MCF-7; a human hepatoma cell line HepG-2; mouse embryonic fibroblast 3T 3; human colon cancer cell lines HT-29; human cervical cancer cell line Hela; canine kidney epithelial cells MDCK; mouse adrenal chromophagous tumor cell PC 12;

experimental drugs: compounds (1-30) of the present invention (prepared as in examples 3-32); ligustrazine (TMP), Cholic Acid (CA), deoxycholic acid (DCA), ursodeoxycholic acid (UCA), hyodeoxycholic acid (HCA), chenodeoxycholic acid (CDCA), lithocholic acid (LCA); the positive drug, Doxorubicin (Doxorubicin) (HY-15142; Haoyuan, Shanghai, Biomedicine technology, Inc.).

2. Method of producing a composite material

2.1 culture of different cell lines

MCF-7, HepG2, 3T3, HT-29, Hela, MDCK and NGF-PC12 cells are cultured in 1640 culture solution containing 10% fetal calf serum and placed at 37 ℃ in 5% CO₂Incubation in an incubator. The cells all grow in an adherent state, the growth condition is observed under an inverted microscope, and subculture is carried out when the number of the cells is proper.

2.2 inhibition of tumor cell proliferation and Normal cell proliferation

Cell testing: MCF-7, HepG2, HT-29, Hela, 3T3, MDCK and NGF-PC12 cells in a logarithmic growth phase are inoculated into a 96-well culture plate in the number of 3 x 103/well, 10 mu L of a compound to be tested or a positive drug is added into each well after the cells are cultured for 4 hours at 37 ℃ in a humidified incubator containing 5% CO2, and the concentration of 9 drugs is tested, so that the final drug concentration is respectively 100, 31.6, 10, 3.16, 1, 0.31, 0.1, 0.03 and 0.01 mu M. Setting a positive control group (100% inhibition) and a negative control group (0% inhibition), repeating the concentration of the drug group for 2 holes, repeating the concentration of the positive control group and the concentration of the negative control group for 6 holes, and carrying out subsequent AlamarBlue test operation after continuously culturing in an incubator for 6 days;

AlamarBlue test procedure: adding 10 mu L of AlamarBlue reagent into each well, incubating for 1-4h, oscillating for 1-2min, and performing enzyme labeling on MD (enzyme labeling) instrument EX: 560nm, EM: measuring fluorescence value at 590nm wavelength, recording the result, calculating the cell inhibition rate (%) of the drug (A0% inhibition-A administration)/(A0% inhibition-A100% inhibition). times.100%, and drawing by using GraphPad Prism 5.0 or MATILAB software and adopting a nonlinear regression method (usually adopting a four-parameter fitting curve equation) to obtain a drug dose response curve, thereby obtaining the IC of the compound₅₀Values and other relevant parameters.

3. Results

3.1 the IC50 values of the compounds of the invention on 5 tumor cell lines (MCF-7, HepG2, HT-29, Hela) and normal cells (3T3, MDCK, NGF-induced PC12) are shown in Table 3.

TABLE 3 IC of different drugs on tumor cell lines and normal cells₅₀Value of

Note that: "-" indicates: according to the result of the cytotoxicity test, when the effect on cancer cells is not obvious, the activity is considered to be low, and the cytotoxicity test on non-cancer cells is not carried out.

As can be seen from Table 3, the compound of the present invention shows proliferation inhibitory activity on various cancer cells, especially compound 10, which has antitumor activity superior to that of the raw material, and antiproliferative activity superior to that of the positive drug adriamycin on human colon cancer cell line HT-29 and human cervical cancer cell line Hela; and the cytotoxicity to NGF-induced PC12 cells, canine kidney epithelial cells MDCK and mouse embryonic fibroblasts 3T3 is obviously lower than that of the positive drug adriamycin.

4. Conclusion

The compound of the invention has better inhibitory activity (IC) on various cancer cells₅₀< 5.7. mu.M), but without any killing effect (IC) on a variety of normal cells at higher doses₅₀More than 80 mu M), has good cytotoxicity selectivity effect.

Experimental example 2 AlamarBlue method for observing the effect of the present invention on proliferation of hepatoma cells and evaluation of response rate in hepatoma

1. Apparatus and materials

Thermo 311 type CO₂An incubator; a Haier biosafety cabinet; molecular Devices microplate reader; hunan instrumental L530 type desk type low-speed centrifuge; olympus IX51 inverted fluorescence microscope, DMEM/F12 medium, fetal bovine serum, 0.25% trypsin solution, phosphate buffer (seimer heishi shanghai ltd); sigma dimethyl sulfoxide (DMSO), resazurin;

liver cancer tissues from independent 9 liver cancer patients were prepared into 9 primary liver cancer cell lines (human liver cancer cell lines HCC3787C1, HCC4172a2, HCC4477B2, HCC4484B1, HCC5850a1, HCC5864D1, HCC5953B1, HCC633a5, and HCC061a2) as follows: fresh liver cancer tissues after surgical resection are taken and cleaned in PBS (GIBCO, 10010-23), are cut into small pieces of 0.5-1mm3 by using instruments such as ophthalmological forceps, scissors and the like in a100 mm culture dish and are laid at the bottom of the dish, 10ml of DMEM/F12 cell culture medium (GIBCO, 11320-containing 033) containing 10% FBS (GIBCO, 10099-containing 141) and 1% Pen-Strep antibiotic (GIBCO, 15140-containing 122) is added into the culture dish, and the culture is carried out in a culture box of 5% CO2 at 37 ℃. The next day, cells were observed under an inverted microscope to migrate from around the tissue mass and then gradually extended to develop into a halo of growth that was visible to the naked eye. After 5-7 days, the histiocytes in the center of the tissue block gradually die and fall off and float, the floating small block can be discarded along with the change of the liquid, and the adherent cells extending from the periphery of the tissue block also gradually form a layer slice. Using the difference in the digestion capacity between fibroblasts and tumor cells, the cells were digested with 0.25% trypsin solution (GIBCO, 25200056) and subcultured to remove fibroblasts continuously. After multiple passages, fibroblasts cannot be observed in the culture dish by naked eyes, and can be used for subsequent experiments when the cells can continuously grow and passages.

Experimental drugs: the compound of the invention (prepared as in example 10); the chemotherapeutic drug Doxorubicin (Doxorubicin) (HY-15142; Shanghai Haoyuan biomedical science, Inc.).

2. Method of producing a composite material

2.1 culture of different cell lines

9 liver cancer cell lines HCC3787C1, HCC4172A2, HCC4477B2, HCC4484B1, HCC5850A1, HCC5864D1, HCC5953B1, HCC633A5 and HCC061A2 are cultured in DMEM/F12 culture solution containing 10% fetal calf serum, placed at 37 ℃ and 5% CO₂Incubation in an incubator. The cells grow in an adherent state, the growth condition is observed under an inverted microscope, and subculture is carried out when the cell confluence rate reaches 80% -90%. The subculture proportion and the number of the hepatoma cell lines are determined according to experimental requirements, and the subculture proportion of the hepatoma cell lines is generally 1: 2-1: 3.

2.2 proliferation inhibition of different hepatoma tumor cell lines

Cell testing: taking 9 liver cancer cell lines HCC3787C1, HCC4172A2, HCC4477B2, HCC4484B1, HCC5850A1, HCC5864D1, HCC5953B1, HCC633A5 and HCC061A2 in logarithmic growth phase, and carrying out reaction at a speed of 1-3 × 10³Per well (optimal density of inoculations per hepatoma cell line determined in preliminary experiments) were plated in 96-well plates in 5% CO₂After 4 hours of culture at 37 ℃ in a humidified incubator, 10. mu.L of the compound of the invention was added to each well, 9 drug concentrations were tested, and the final drug concentrations were made to be 100, 31.6, 10, 3.1, 1, 0.3, 0.1, 0.03, and 0.01. mu.M, respectively, and a QC reference compound, Doxorubicin, was added simultaneously at the time of testing each hepatoma cell line, and the final drug concentrations were 10, 3.16, 1, 0.31, 0.1, 0.03, 0.01, 0.003, and 0.001. mu.M, respectively. In addition, a positive control group (100% inhibition) and a negative control group (0% inhibition) are simultaneously arranged, 2 holes are repeated for each concentration of the medicine group, 6 holes are repeated for the positive control group and the negative control group, and after the positive control group and the negative control group are continuously cultured in an incubator for 6 days, the subsequent AlamarBlue test operation is carried out;

AlamarBlue test procedure: adding 10 mu L of AlamarBlue reagent into each well, incubating for 1-4h, oscillating for 1-2min, and performing enzyme labeling on MD (enzyme labeling) instrument EX: 560nm, EM: the fluorescence value was measured at 590nm wavelength, and the results were recorded, and the cell inhibition ratio (%) of the compound of the present invention was calculated as (A)_{0% inhibition}-A_{Administration of drugs})/(A_{0% inhibition}-A_{100% inhibition}) X 100%, and then drawing by using GraphPad Prism 5.0 or MATILAB software and adopting a nonlinear regression method (usually adopting a four-parameter fitting curve equation) to obtain a drug dose response curve, thereby obtaining the IC of the compound acting on 9 hepatoma cell lines₅₀The value is obtained.

3. Results

3.1 IC of Compound 10 of the present invention against 9 liver cancer cell lines (HCC3787C1, HCC4172A2, HCC4477B2, HCC4484B1, HCC5850A1, HCC5864D1, HCC5953B1, HCC633A5 and HCC061A2)₅₀The values are shown in Table 4.

TABLE 4 IC of Compound 10 of the present invention against 9 hepatoma cell lines₅₀Value of

As can be seen from Table 4, Compound 10 of the present invention has a better inhibitory activity (IC) against 7 liver cancer cell lines (HCC4477B2, HCC4484B1, HCC5850A1, HCC5864D1, HCC5953B1, HCC633A5 and HCC061A2)₅₀< 10. mu.M), inhibitory activity IC against 2 liver cancer cell lines HCC4172A2 and HCC3787C1₅₀About 10 μ M. Therefore, the compound has good effect of inhibiting the proliferation of the liver cancer cells and has good response rate in the liver cancer cells

Experimental example 3 anti-tumor Effect of Compound 10 of the present invention on H22 sarcoma mouse

1. Material

1.1 Experimental animals and tumor strains:

healthy male ICR mice, weighing 18-22g, were purchased from the laboratory animal center of Wintolite laboratory animal technology corporation (certification code SCXK (Jing) 2011-; mouse sarcoma cell H22 was obtained as a gift from the university of Western Ann traffic. Tumor-bearing H22 mice, passaged every 5 days.

1.2 Experimental drugs:

the purity of the compound 10 is determined by liquid chromatography (HPLC) analysis, and is more than or equal to 98 percent, thereby meeting the experimental requirements. The powder was stored sealed at 4 ℃.

Cyclophosphamide for injection: shanxi Pude pharmaceutical Co., Ltd (production batch: 04140202)

2. Method of producing a composite material

2.1 model animal construction

Killing inoculated 7-day H22 mouse by breaking neck, respectively taking ascites under aseptic condition, washing with RPMI1640 culture medium for 2 times, and making into 2 × 10 with sterile physiological saline⁷The cell suspension/ml, H22 cell suspension was inoculated under the skin of the right axilla of mice, 0.1ml each, 72 were inoculated, and solid tumor models were prepared.

2.2 Experimental groups

The 72 mice were randomly divided into 6 groups by weight, namely a normal control group, a positive drug (cyclophosphamide) group, a model group, a low dose group, a medium dose group and a high dose group of the compound of the present invention; each group had 12.

2.3 methods of administration

The compound 10 is prepared from soybean oil for injection, the dosage of a low-dose group is 10mg/kg, the dosage of a medium-dose group is 30mg/kg, the dosage of a high-dose group is 90mg/kg, a negative control group is injected with soybean oil for injection with equal volume, a positive control group is injected with cyclophosphamide injection 30mg/kg, every other day is injected with abdominal cavity, and the continuous administration is carried out for 12 days. During the period, the mice were observed daily for general activities, fur, feces, body weight, and the like. 24H after the last dose, mice vaccinated subcutaneously with H22 were sacrificed by cervical amputation and tumor, liver, spleen and kidney were removed.

2.4 calculation of visceral index

After the administration of all the solid tumor groups, the mice were weighed and then sacrificed, and the spleen, kidney and liver were weighed using an electronic balance, respectively. The spleen index is the weight of spleen (mg)/weight of mouse (g) of each group of mice, the liver index is the weight of liver (100 g)/weight of mouse (g), and the kidney index is the weight of kidney (mg)/weight of mouse (g).

2.5 calculation of tumor inhibition Rate

The solid tumor negative control group is weighed the next day after drug withdrawal, (eyeball bleeding is removed, serum is collected for subsequent experiments) the mice are killed after being taken out of the neck, tumor tissues are dissected, weighing is carried out by an electronic balance, and the tumor inhibition rate is calculated.

2.6 statistical treatment

All data are statistically analyzed by SPSS11.0 software package, and the comparison of counting data is performed by x²And (6) checking. P is less than 0.05, which has statistical significance.

3. Results

3.1 in vivo anticancer Activity of Compound 10 of the present invention

The results are shown in Table 5.

TABLE 5 inhibitory Effect of Compound 10 of the present invention on H22 tumor-bearing mice (x. + -. s)

Note: comparison with normal controls: p < 0.05, P < 0.01

As shown in Table 5, the compound 10 has no influence on the weight gain of H22 sarcoma mice, and has obvious tumor inhibiting effect, the tumor growth inhibiting rates of the low, medium and high dose groups to H22 sarcoma mice are 46.09%, 60.16% and 50.78%, respectively, and the tumor inhibiting effect of the high dose group is equivalent to that of the positive drug cyclophosphamide.

3.2 changes in liver, spleen and Kidney indices of various groups of H22 mice

The results are shown in Table 6.

TABLE 6 Effect of Compound 10 of the present invention on the liver, spleen and Kidney indices (x + s) of H22 mice

Note: comparison with blank control group: p < 0.05, P < 0.01

As shown in Table 6, compared with the normal control group, the liver-spleen index of the model group is increased, the spleen index change is particularly obvious (P is less than 0.05), and the kidney index is reduced. After treatment, cyclophosphamide and the compound can reduce the liver and spleen indexes of H22 sarcoma mice to be close to normal levels.

4. Conclusion

4.1 Compound 10 of the present invention significantly inhibited the growth of H22 mouse sarcoma.

4.2 Compound 10 of the present invention modulates the liver and spleen index of H22 sarcoma mice.

Experimental example 4 acute toxicity test of Compound 10 of the present invention

1. Material

1.1 Experimental animals

Healthy ICR mice, each male and female 12, with a weight of 18-22g, purchased from the laboratory animal center of Wintonlihua laboratory animal technology company (certificate number SCXK (Jing) 2006-0009)

1.2 Experimental drugs

The purity of the compound 10 is determined by liquid chromatography (HPLC) analysis, and is more than or equal to 98 percent, thereby meeting the experimental requirements. The powder was stored sealed at 4 ℃. Dissolving 450mg of the compound in appropriate amount of ethyl acetate, mixing with 5.0ml of soybean oil for injection, and ultrasonic suspending, and recovering ethyl acetate as solvent until it is odorless. It is prepared as before use.

2. Method of producing a composite material

40 mice were randomly divided into 4 groups of 10 mice each based on body weight, the experimental group was administered by intraperitoneal injection of mice, and the control group was administered with the same amount of soybean oil for injection. Mice were fasted overnight (12 hours continuously) prior to dosing, and at 9 am of the day: 00 (900mg/kg) was started and was fed after 6 hours of continuous observation for 14 days.

3. Results

After the intraperitoneal injection administration (900mg/kg), slight writhing phenomenon appears in individual mice, symptoms disappear within 1 hour approximately, death is not seen within 12 hours after the administration, and no obvious toxic and side effects are seen in blank groups. After 14 days, the state of each group of mice is good, no death is seen, and no abnormal change is seen when visceral organs such as heart, liver, spleen, lung, kidney, adrenal gland, thyroid gland and the like are observed after the cervical vertebra dislocation of the mice is died. Acute toxicity test of compound 26 injected in abdominal cavity of mouse shows that the medicine LD₅₀＞900mg/kg。

4. Conclusion

Compound 10 of the present invention is injected into normal mouse abdominal cavity with LD₅₀More than 900mg/kg, and higher safety.

Detailed Description

EXAMPLE 12 preparation of bromomethyl-3, 5, 6-trimethylpyrazine (intermediate Compound 1)

According to optimization of a synthesis process of an intermediate 2-bromomethyl-3, 5, 6-trimethyl pyrazine, introduction, Rongpenlong, Hanqijun, and the like, Anhui medicine, 2013, 17 (9): 1467-1470' method. Weighing 20.00g (0.15mol) of anhydrous ligustrazine and 23.54g of NBS N-bromosuccinimide (0.15mol, ground before use) in a 250mL three-necked flask, 100mL CCl₄As a reaction solvent, 4 incandescent lamps of 85W were used for irradiation, and the reaction was refluxed at 95 ℃ for 1 hour. TLC (3: 1 ratio of V (petroleum ether) to V (acetone))]The detection reaction is substantially complete; cooling, filtering, collecting filtrate, and recovering solvent under reduced pressure to obtain mauve viscous liquid (content is 60%). HRMS (ESI) m/z: 216.00135[ M + H]⁺，calcd.for C₈H₁₁BrN₂ 216.00851。

Example 23 preparation of 5, 6-trimethylpyrazine-2-carboxylic acid (intermediate Compound 2)

According to "the synthesis of novel ligustrazine derivatives and the study of their anticancer activity", journal of northwest pharmacology, 29 (1): 58-64' method. 10.0g (73.53mmol) of ligustrazine is suspended in 100.0mL of distilled water, 11.62g (73.53mmol) of potassium permanganate is weighed and added in 3 batches, and the mixture is magnetically stirred at 37 ℃ for 24 hours. Cooling the mixed solution, performing suction filtration, adjusting the pH of the filtrate to 1-2 by using 360 mg.L < -1 > hydrochloric acid, extracting for 3 times by using ethyl acetate, collecting an ethyl acetate layer, drying by using anhydrous sodium sulfate, concentrating under reduced pressure to dryness, and recrystallizing by using acetone to obtain a white solid, wherein the yield is as follows: 47.8%, m.p.: 162 ℃ and 163 ℃.

EXAMPLE 3 preparation of Compound 1

Weighing 2.19mmol of 2-bromomethyl-3, 5, 6-trimethyl pyrazine and 2.19mmol of deoxycholic acid, placing the materials in a 50ml round-bottom flask, adding 25ml of DMF, adding 3mmol of potassium carbonate after the mixture is dissolved, stirring at 85 ℃ for 2h, monitoring by TLC that the reaction raw materials basically disappear, stopping the reaction, adding a large amount of saturated NaCl solution into the reaction solution for dispersion, extracting by 300ml of ethyl acetate twice, and using anhydrous Na for an ethyl acetate layer₂SO₄Drying, concentrating under reduced pressure, dissolving the residue with 3ml chloroform, adding 2.8g silica gel, evaporating under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 8: 1 to obtain white solid 0.594 g. m.p.: 83.6-84.3 ℃, yield 58.4%. HRMS (ESI) m/z: 527.5201[ M + H]⁺，calcd.for C₃₂H₅₁N₂O₄ 527.3849.

EXAMPLE 4 preparation of Compound 2

Weighing 11.2mmol of compound and 1.8mmol of ligustrazine acid, placing in a 50ml round bottom flask, adding 25ml of CH₂Cl₂After the mixture is dissolved, EDCI 1.8mmol and DMAP 0.18mmol are respectively added, the mixture is stirred for 12h at room temperature, TLC monitors that the reaction raw materials basically disappear, the reaction is stopped, the mixture is concentrated under reduced pressure, 2.2g of silica gel is added, the mixture is evaporated to dryness under reduced pressure and is mixed with a sample, and an eluant is petroleum ether and acetone which are 7: 1 to elute, so that 0.426g of white solid is obtained. White Amorphous solid, m.p.: 60.5-61.2 ℃ and yield 52.7 percent.¹H-NMR(CDCl₃)(ppm)：0.67(s，3H，H-18)，0.96(s，3H，H-19)，0.97(d，J＝ 7.0Hz，3H，H-21)，3.99(m，1H，H-12)，5.04(m，1H，H-3)，5.19(t，J＝12.5 Hz，2H，CH₂-2′)，2.72(s，3H，CH₃-6″)，2.57(s，3H，CH₃-5″)，2.56(s，3H， CH₃-3″)，2.54(s，3H，CH₃-6′)，2.52(s，3H，CH₃-5′)，2.51(s，3H，CH₃-3′)， 1.00-2.50(27H，methyl-and methylene-of steroid structure).¹³C-NMR (CDCl3)(ppm)：35.0(C1)，26.5(C2)，76.2(C3)，32.1(C4)，42.0(C5)，27.0 (C6)，26.0(C7)，36.0(C8)，33.7(C9)，34.2(C10)，28.7(C11)，73.1(C12)， 46.5(C13)，48.2(C14)，23.6(C15)，27.4(C16)，47.3(C17)，12.7(C18)，23.1 (C19)，35.1(C20)，17.3(C21)，31.1(C22)，30.9(C23)，173.8(C24)；pyrazine ring：64.9(2′-CH₂)，151.2(C2′)，144.9(C3′)，148.9(C5′)，149.1(C6′)，166.8 (2″-C＝O)，154.5(C2″)，140.4(C3″)，149.3(C5″)，150.5(C6″)，21.6(6′-CH₃)， 21.4(5′-CH₃)，20.4(3′-CH₃)，22.5(6″-CH₃)，22.1(5″-CH₃)，21.6(3″-CH₃). HRMS(ESI)m/z：675.44635[M+H]⁺，calcd.for C₄₀H₅₈N₄O₅ 674.44072.

Preparation of intermediate deoxycholic acid benzyl ester: weighing benzyl bromide 5.0mmol and deoxycholic acid 5.0mmol, placing in 50ml round bottom flask, adding 25ml DMF, dissolving the mixture, adding 5mmol potassium carbonate, stirring at 85 deg.C for 2h, monitoring by TLC that the reaction raw materials basically disappear, stopping the reaction, adding a large amount of saturated NaCl solution into the reaction solution for dispersion, extracting twice with 400ml ethyl acetate, and using anhydrous Na for ethyl acetate layer₂SO₄Drying, concentrating under reduced pressure, dissolving the residue with 5ml chloroform, adding 3.8g silica gel, evaporating under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 5: 1 to obtain white solid 2.212 g. White powder, m.p.: 136.5-137.3 ℃ and yield 91.8 percent.

EXAMPLE 5 preparation of Compound 3

Weighing deoxycholic acid benzyl ester 3.0mmol, 4.0mmol of ligustrazine acid are placed in a 50ml round bottom flask, 30ml of CH is added₂Cl₂After the mixture is dissolved, respectively adding EDCI 4.0mmol and DMAP 0.4mmol, stirring at room temperature for 12h, monitoring by TLC that the reaction raw material basically disappears, stopping the reaction, washing the reaction solution twice by 60ml of saturated NaCl solution, and obtaining CH₂Cl₂The layer was dried over anhydrous Na2SO4, concentrated under reduced pressure, added with 3.2g silica gel, evaporated to dryness under reduced pressure and mixed with the eluent petroleum ether and acetone at 7: 1 to give 0.924g white solid. White Amorphous solid, m.p.: 47.1-48.0 ℃ and yield 48.9 percent.¹H-NMR(CDCl₃)(ppm)：0.67(s，3H，H-18)，0.97(s，3H，H-19)，0.98 (d，J＝7.5Hz，3H，H-21)，3.99(m，1H，H-12)，5.05(m，1H，H-3)，5.14，5.11 (ea，d，J＝12.5Hz，1H，CH₂-1′)，7.37(m，5H，H-2′，3′，4′，5′，6′)，2.72(s，3H， CH₃-6″)，2.58(s，3H，CH₃-5″)，2.57(s，3H，CH₃-3″)，1.00-2.50(27H，methyl- and methylene-of steroid structure).¹³C-NMR(CDCl₃)(ppm)：35.0(C1)， 26.5(C2)，76.2(C3)，32.3(C4)，42.1(C5)，27.0(C6)，26.1(C7)，36.0(C8)， 33.7(C9)，34.2(C10)，28.7(C11)，73.1(C12)，46.5(C13)，48.2(C14)，23.6 (C15)，27.4(C16)，47.5(C17)，12.7(C18)，23.1(C19)，35.0(C20)，17.3 (C21)，31.3(C22)，30.9(C23)，174.0(C24)，66.1(1′-CH₂)，136.1(C1′)，128.2 (C2′，6′)，128.5(C3′，5′)，128.2(C4′)；pyrazine ring：165.8(2″-C＝O)，153.9 (C2″)，140.5(C3″)，149.2(C5″)，150.4(C6″)，22.5(6″-CH₃)，22.1(5″-CH₃)， 21.6(3″-CH₃).HRMS(ESI)m/z：653.39187[M+Na]⁺，calcd.for C₃₉H₅₄N₂O₅630.40327.

EXAMPLE 6 preparation of Compound 4

Weighing 31.2mmol of compound, placing in a 50ml round bottom flask, adding 30ml THF, after the mixture is dissolved, adding 75mg of Pd/C, reducing by hydrogen, stirring at room temperature for 8h, monitoring by TLC that the reaction raw materials are basically disappeared, stopping the reaction, filtering to remove Pd/C, concentrating the reaction solution under reduced pressure, and using 3ml of CHCl for the residue₃Redissolving, adding 2.2g of silica gel, evaporating to dryness under reduced pressure, mixing the sample, and eluting with petroleum ether and acetone at a ratio of 5: 1 to obtain 0.673g of white solid. White Amorphous solid, m.p.: 182.8-183.5 ℃ and yield 58.4 percent.¹H-NMR(CDCl₃)(ppm)：0.69(s，3H，H-18)，0.96(s，3H，H-19)， 0.98(d，J＝6.0Hz，3H，H-21)，4.01(m，1H，H-12)，5.04(m，1H，H-3)，2.73(s， 3H，CH₃-6′)，2.57(s，3H，CH₃-5′)，2.57(s，3H，CH₃-3′)，1.00-2.50(28H， methyl-and methylene-of steroid structure).¹³C-NMR(CDCl₃)(ppm)：35.0 (C1)，26.5(C2)，76.2(C3)，32.1(C4)，42.1(C5)，27.0(C6)，26.0(C7)，36.0 (C8)，33.7(C9)，34.2(C10)，28.7(C11)，73.2(C12)，46.5(C13)，48.2(C14)， 23.6(C15)，27.5(C16)，47.3(C17)，12.7(C18)，23.1(C19)，35.1(C20)，17.3 (C21)，31.0(C22)，30.7(C23)，179.0(C24).pyrazine ring：165.6(2′-C＝O)， 154.1(C2′)，140.3(C3′)，149.3(C5′)，150.6(C6′)，22.4(6′-CH₃)，22.0 (5′-CH₃)，21.4(3′-CH₃).HRMS(ESI)m/z：541.36188[M+H]⁺，calcd.for C₃₂H₄₈N₂O₅ 540.35632.

EXAMPLE 7 preparation of Compound 5

Weighing 2.19mmol of 2-bromomethyl-3, 5, 6-trimethyl pyrazine and 2.19mmol of cholic acid, placing in a 50ml round bottom flask, adding 25ml of DMF, dissolving the mixture, adding 3mmol of potassium carbonate, stirring at 85 ℃ for 2h, monitoring by TLC that the reaction raw materials basically disappear, stopping the reaction, adding a large amount of saturated NaCl solution into the reaction solution for dispersion, extracting twice with 300ml of ethyl acetate, drying the ethyl acetate layer with anhydrous Na2SO4, concentrating under reduced pressure, dissolving the residue with 3ml of chloroform, adding 2.8g of silica gelEvaporating to dryness under reduced pressure, and mixing with eluent petroleum ether and acetone at a ratio of 8: 1 to obtain white solid 0.594 g. m.p.: 95.4-96.3 ℃, yield 55.4%. HRMS (ESI) m/z: 543.6199[ M + H]⁺，calcd.for C₃₂H₅₁N₂O₅ 543.3798.

EXAMPLE 8 preparation of Compound 6

Weighing 51.2mmol of compound and 1.8mmol of ligustrazine acid, placing in a 50ml round bottom flask, adding 25ml of CH₂Cl₂After the mixture is dissolved, EDCI 1.8mmol and DMAP 0.18mmol are respectively added, the mixture is stirred for 12h at room temperature, TLC monitors that the reaction raw materials basically disappear, the reaction is stopped, the mixture is concentrated under reduced pressure, 2.2g of silica gel is added, the mixture is evaporated to dryness under reduced pressure and is mixed, and an eluent is petroleum ether and acetone which are 7: 1 to elute, so that 0.396g of white solid is obtained. White Amorphous solid, m.p.: 85.7-86.5 ℃ and yield 47.8 percent.¹H-NMR(CDCl3)(ppm)：0.68(s，3H，H-18)，0.93(s，3H，H-19)，0.96(d，J＝ 5.5Hz，3H，H-21)，4.90(m，1H，H-3)，3.87(m，1H，H-7)，3.98(m，1H，H-12)， 5.19(brs，2H，CH₂-2′)，2.71(s，3H，CH₃-6″)，2.56(s，3H，CH₃-5″)，2.56(s，3H， CH₃-3″)，2.54(s，3H，CH₃-6′)，2.52(s，3H，CH₃-5′)，2.51(s，3H，CH₃-3′)， 1.00-2.50(26H，methyl-and methylene-of steroid structure).¹³C-NMR (CDCl₃)(ppm)：35.1(C1)，26.7(C2)，76.2(C3)，34.9(C4)，41.3(C5)，34.8 (C6)，68.2(C7)，39.5(C8)，26.6(C9)，34.4(C10)，28.4(C11)，72.9(C12)， 46.5(C13)，42.0(C14)，23.1(C15)，27.4(C16)，47.1(C17)，12.5(C18)，22.5 (C19)，35.2(C20)，17.3(C21)，31.0(C22)，30.8(C23)，173.9(C24)；pyrazine ring：64.9(2′-CH₂)，151.2(C2′)，144.9(C3′)，148.9(C5′)，149.1(C6′)，165.8 (2″-C＝O)，153.9(C2″)，140.5(C3″)，149.2(C5″)，150.4(C6″)，21.5(6′-CH₃)， 21.4(5′-CH₃)，20.4(3′-CH₃)，22.6(6″-CH₃)，22.1(5″-CH₃)，21.6(3″-CH₃). HRMS(ESI)m/z：691.44092[M+H]⁺，calcd.for C₄₀H₅₈N₄O₆690.43564.

Preparation of intermediate benzyl cholate: weighing benzyl bromide 5.0mmol and cholic acid 5.0mmol, placing in 50ml round bottom flask, adding 25ml DMF, dissolving the mixture, adding 5mmol potassium carbonate, stirring at 85 deg.C for 2 hr, monitoring by TLC that reaction raw materials basically disappear, stopping reaction, adding a large amount of saturated NaCl solution to the reaction solution for dispersion, extracting with 400ml ethyl acetate twice, and using anhydrous Na for ethyl acetate layer₂SO₄Drying, concentrating under reduced pressure, dissolving the residue with 5ml chloroform, adding 3.8g silica gel, evaporating under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 5: 1 to obtain 2.243g white solid. White powder, m.p.: 177.8-178.6 ℃ and yield 90.1 percent.

EXAMPLE 9 preparation of Compound 7

Weighing 3.0mmol of intermediate benzyl cholic acid and 4.0mmol of ligustrazine acid, placing in a 50ml round bottom flask, adding 30ml CH₂Cl₂After the mixture is dissolved, respectively adding EDCI 4.0mmol and DMAP 0.4mmol, stirring at room temperature for 12h, monitoring by TLC that the reaction raw material basically disappears, stopping the reaction, washing the reaction solution twice by 60ml of saturated NaCl solution, and obtaining CH₂Cl₂Anhydrous Na for layer₂SO₄Drying, concentrating under reduced pressure, adding 3.2g silica gel, evaporating to dryness under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 7: 1 to obtain white solid 0.928 g. White Amorphous solid, m.p.: 169.0-169.6 ℃ and yield 47.9 percent.¹H-NMR(CDCl₃)(ppm)：0.69(s，3H，H-18)，0.95(s，3H，H-19)，0.98 (d，J＝6.0Hz，3H，H-21)，4.91(m，1H，H-3)，3.88(m，1H，H-7)，4.00(m，1H， H-12)，5.15，5.11(ea，d，J＝12.5Hz，1H，CH₂-1′)，7.37(m，5H，H-2′，3′，4′，5′， 6′)，2.72(s，3H，CH₃-6″)，2.57(s，3H，CH₃-5″)，2.57(s，3H，CH₃-3″)， 1.00-2.50(26H，methyl-and methylene-of steroid structure).¹³C-NMR (CDCl3)(ppm)：35.1(C1)，26.8(C2)，76.2(C3)，34.9(C4)，41.3(C5)，34.8 (C6)，68.2(C7)，39.6(C8)，26.7(C9)，34.4(C10)，28.4(C11)，72.9(C12)， 46.5(C13)，42.0(C14)，23.1(C15)，27.4(C16)，47.2(C17)，12.5(C18)，22.5 (C19)，35.1(C20)，17.3(C21)，31.3(C22)，30.8(C23)，174.0(C24)，66.1 (1′-CH2)，136.1(C1′)，128.2(C2′，6′)，128.5(C3′，5′)，128.2(C4′)；pyrazine ring：165.8(2″-C＝O)，153.9(C2″)，140.6(C3″)，149.3(C5″)，150.4(C6″)， 22.6(6″-CH₃)，22.1(5″-CH₃)，21.6(3″-CH₃).HRMS(ESI)m/z：647.40668 [M+H]⁺，calcd.for C₃₉H₅₄N₂O₆ 646.39819.

EXAMPLE 10 preparation of Compound 8

Weighing 71.2mmol of compound, placing in 50ml round bottom flask, adding 30ml THF, dissolving the mixture, adding 78mg Pd/C, reducing with hydrogen, stirring at room temperature for 10h, monitoring by TLC that the reaction material is substantially disappeared, stopping the reaction, filtering to remove Pd/C, concentrating the reaction solution under reduced pressure, and adding 3ml CHCl into the residue₃Redissolving, adding 2.2g of silica gel, evaporating to dryness under reduced pressure, mixing the sample, and eluting with petroleum ether and acetone at a ratio of 5: 1 to obtain 0.590g of white solid. White Amorphous solid, m.p.: 132.6-133.5 deg.C, yield 88.4%. 1H-NMR (CDCl)₃)(ppm)：0.70(s，3H，H-18)，0.93(s，3H，H-19)， 0.98(d，J＝5.5Hz，3H，H-21)，4.89(m，1H，H-3)，3.88(m，1H，H-7)，4.00(m， 1H，H-12)，2.73(s，3H，CH₃-6′)，2.57(s，3H，CH₃-3′，5′)，1.00-2.50(27H， methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：35.1 (C1)，26.7(C2)，76.3(C3)，34.9(C4)，41.3(C5)，34.8(C6)，68.3(C7)，39.5 (C8)，26.6(C9)，34.4(C10)，28.3(C11)，73.0(C12)，46.5(C13)，41.9(C14)，23.2(C15)，27.5(C16)，47.0(C17)，12.5(C18)，22.5(C19)，35.2(C20)，17.3 (C21)，30.9(C22)，30.7(C23)，178.3(C24)；pyrazine ring：165.6(2′-C＝O)， 154.0(C2′)，140.3(C3′)，149.2(C5′)，150.6(C6′)，22.5(6′-CH₃)，22.0 (5′-CH₃)，21.3(3′-CH₃).HRMS(ESI)m/z：555.34312[M-H]^-，calcd.for C₃₂H₄₈N₂O₆ 556.35124.

EXAMPLE 11 preparation of Compound 9

Weighing 2.19mmol of 2-bromomethyl-3, 5, 6-trimethylpyrazine and 2.19mmol of hyodeoxycholic acid, placing the mixture in a 50ml round-bottom flask, adding 25ml of DMF, adding 3mmol of potassium carbonate after the mixture is dissolved, stirring at 85 ℃ for 2h, monitoring by TLC that reaction raw materials basically disappear, stopping reaction, adding a large amount of saturated NaCl solution into a reaction solution for dispersion, extracting by 300ml of ethyl acetate twice, and using anhydrous Na for an ethyl acetate layer₂SO₄Drying, concentrating under reduced pressure, dissolving the residue with 3ml chloroform, adding 2.8g silica gel, evaporating under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 8: 1 to obtain white solid 0.594 g. m.p.: 65.3-66.1 deg.C, yield 51.6%. 1H-NMR (CDCl)₃)(ppm)：0.62(s，3H， H-18)，0.90(s，3H，H-19)，0.91(d，J＝6.0Hz，3H，H-21)，3.61(m，1H，H-3)， 4.05(m，1H，H-6)，5.19(t，J＝12.5Hz，2H，CH₂-2′)，2.53(s，3H，CH₃-6′)， 2.51(s，3H，CH₃-5′)，2.51(s，3H，CH₃-3′)，1.00-2.50(28H，methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：35.6(C1)，29.2 (C2)，71.5(C3)，28.1(C4)，48.5(C5)，68.0(C6)，35.0(C7)，34.8(C8)，39.8 (C9)，35.9(C10)，20.7(C11)，39.9(C12)，42.8(C13)，56.1(C14)，24.2(C15)， 30.2(C16)，55.9(C17)，12.0(C18)，23.5(C19)，35.3(C20)，18.2(C21)，31.1 (C22)，30.9(C23)，173.9(C24)；pyrazine ring：64.9(2′-CH₂)，151.2(C2′)， 144.9(C3′)，149.0(C5′)，149.0(C6′)，21.6(6′-CH₃)，21.4(5′-CH₃)，20.5 (3′-CH₃).HRMS(ESI)m/z：527.38214[M+H]⁺，calcd.for C₃₂H₅₀N₂O₄ 526.37706.

EXAMPLE 12 preparation of Compound 10

Weighing about 1.2mmol of compound 9 and 1.8mmol of ligustrazine acid, placing in a 50ml round bottom flask, adding 25ml of CH₂Cl₂After the mixture is dissolved, respectively adding EDCI 1.8mmol and DMAP 0.18mmol, stirring at room temperature for 12h, monitoring by TLC that the reaction raw material basically disappears, stopping the reaction, concentrating under reduced pressure, adding 2.2g of silica gel, evaporating to dryness under reduced pressure, mixing with a sample, and eluting with petroleum ether and acetone which are 8: 1, and a white semisolid which is 0.181 g.m.p.: 72.3-73.1 deg.C, yield 18.4%. 1H-NMR (CDCl)₃) (ppm)：0.67(s，3H，H-18)，1.01(s，3H，H-19)，0.92(d，J＝6.5Hz，3H，H-21)， 5.04(m，1H，H-3)，5.51(m，1H，H-6)，5.20(t，J＝12.5Hz，2H，CH2-2′)， 2.48-2.71(s，each，3H，9×CH₃-pyrazine ring)，1.00-2.50(25H，methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：35.3(C1)，26.5 (C2)，75.5(C3)，26.4(C4)，45.6(C5)，72.9(C6)，31.3(C7)，34.8(C8)，39.9 (C9)，36.3(C10)，20.7(C11)，39.9(C12)，42.9(C13)，56.1(C14)，24.1(C15)， 28.1(C16)，55.9(C17)，12.0(C18)，23.3(C19)，35.1(C20)，18.2(C21)，31.1 (C22)，30.9(C23)，173.8(C24)；pyrazine ring：65.0(2′-CH2)，151.2(C2′)， 144.9(C3′)，149.0(C5′)，149.0(C6′)，165.5(2″-C＝O)，154.1(C2″)，140.4 (C3″)，149.3(C5″)，150.2(C6″)，165.7(2″′-C＝O)，154.0(C2″′)，140.3(C3″′)， 149.2(C5″′)，150.5(C6″)，21.6(6′-CH₃)，21.5(5′-CH₃)，20.5(3′-CH₃)，22.6 (6″-CH₃)，22.1(5″-CH₃)，21.7(3″-CH₃)，22.5(6″′-CH₃)，22.1(5″′-CH₃)，21.6 (3″′-CH₃).HRMS(ESI)m/z：823.51044[M+H]⁺，calcd.for C₄₈H₆₆N₆O₆ 822.50438.

EXAMPLE 13 preparation of Compound 11

This structure was isolated in the preparation of compound 10 as a white semi-solid, 0.149 g. Semi solid, yield 22.1%. 1H-NMR (CDCl)₃)(ppm)：0.66(s，3H，H-18)，1.04(s， 3H，H-19)，0.92(d，J＝6.5Hz，3H，H-21)，3.61(m，1H，H-3)，5.47(m，1H， H-6)，5.20(t，J＝12.5Hz，2H，CH₂-2′)，2.71(s，3H，CH₃-6″)，2.56(brs，6H， CH₃-3″，5″)，2.54(s，3H，CH₃-6′)，2.52(s，3H，CH₃-5′)，2.52(s，3H，CH₃-3′)， 1.00-2.50(27H，methyl-and methylene-of steroid structure).13C-NMR (CDCl₃)(ppm)：35.4(C1)，30.2(C2)，71.2(C3)，28.1(C4)，45.5(C5)，73.3 (C6)，31.3(C7)，34.8(C8)，39.9(C9)，36.2(C10)，20.7(C11)，39.9(C12)， 42.9(C13)，56.1(C14)，24.1(C15)，30.3(C16)，55.9(C17)，12.0(C18)，23.3 (C19)，35.3(C20)，18.2(C21)，31.1(C22)，30.9(C23)，173.8(C24)；pyrazine ring：64.9(2′-CH2)，151.2(C2′)，144.9(C3′)，149.0(C5′)，149.0(C6′)，165.7 (2″-C＝O)，154.0(C2″)，140.4(C3″)，149.3(C5″)，150.2(C6″)，21.6(6′-CH₃)， 21.4(5′-CH₃)，20.5(3′-CH₃)，22.5(6″-CH₃)，22.1(5″-CH₃)，21.6(3″-CH₃). HRMS(ESI)m/z：675.44873[M+H]⁺，calcd.for C₄₀H₅₈N₄O₅ 674.44072.

Preparation of hyodeoxycholic acid benzyl ester: placing benzyl bromide 5.0mmol and hyodeoxycholic acid 5.0mmol in 50ml round bottom flask, adding 25ml DMF, dissolving the mixture, adding 5mmol potassium carbonate, stirring at 85 deg.C for 2 hr, monitoring by TLC that reaction raw materials basically disappear, stopping reaction, adding a large amount of saturated NaCl solution to the reaction solution for dispersion, extracting with 400ml ethyl acetate twice, and using anhydrous Na for ethyl acetate layer₂SO₄Drying, concentrating under reduced pressure, dissolving the residue with 5ml chloroform, adding 3.8g silica gel, evaporating under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 5: 1 to obtain white solid 2.234 g. White powder, m.p.: 132.4-133.3 ℃ and yield 92.7 percent.

EXAMPLE 14 preparation of Compound 12

3.0mmol of hyodeoxycholic acid benzyl ester and 4.0mmol of ligustrazine acid are weighed and placed in a 50ml round bottom flask, and 30ml of CH is added₂Cl₂After the mixture is dissolved, respectively adding EDCI 4.0mmol and DMAP 0.4mmol, stirring at room temperature for 12h, monitoring by TLC that the reaction raw material basically disappears, stopping the reaction, washing the reaction solution twice by 60ml of saturated NaCl solution, and obtaining CH₂Cl₂Anhydrous Na for layer₂SO₄Drying, concentrating under reduced pressure, adding 3.2g silica gel, evaporating to dryness under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 7: 1 to obtain white solid 0.391 g. M.p.: 55.3-56.1 deg.C, yield 20.7%. 1H-NMR (CDCl)₃) (ppm)：0.66(s，3H，H-18)，1.04(s，3H，H-19)，0.92(d，J＝6.5Hz，3H，H-21)， 3.64(m，1H，H-3)，5.48(m，1H，H-6)，5.14，5.11(ea，d，J＝12.5Hz，1H， CH₂-1′)，7.37(m，5H，H-2′，3′，4′，5′，6′)，2.71(s，3H，CH3-6″)，2.57(brs，6H， CH₃-3″，5″)，1.00-2.50(26H，methyl-and methylene-of steroid structure). 13C-NMR(CDCl₃)(ppm)：35.4(C1)，30.2(C2)，71.2(C3)，28.1(C4)，45.5 (C5)，73.3(C6)，31.3(C7)，34.8(C8)，39.9(C9)，36.2(C10)，20.7(C11)，39.9 (C12)，42.9(C13)，56.1(C14)，24.2(C15)，30.2(C16)，55.9(C17)，12.0 (C18)，23.4(C19)，35.3(C20)，18.2(C21)，31.0(C22)，30.9(C23)，174.1 (C24)，66.1(1′-CH₂)，136.1(C1′)，128.3(C2′，6′)，128.6(C3′，5′)，128.2(C4′)； pyrazine ring：165.7(2″-C＝O)，154.0(C2″)，140.4(C3″)，149.4(C5″)，150.2 (C6″)，22.6(6″-CH₃)，22.1(5″-CH₃)，21.7(3″-CH₃).HRMS(ESI)m/z：631.40991[M+H]⁺，calcd.for C₃₉H₅₄N₂O₅ 630.40327.

EXAMPLE 15 preparation of Compound 13

This structure was isolated in the preparation of compound 12 as a white solid, 0.448 g.m.p.: 61.3-62.1 deg.C, yield 19.2%. 1H-NMR (CDCl)₃)(ppm)：0.66(s，3H，H-18)， 1.09(s，3H，H-19)，0.92(d，J＝6.5Hz，3H，H-21)，5.04(m，1H，H-3)，5.50(m， 1H，H-6)，5.14，5.11(ea，d，J＝12.0Hz，2H，CH₂-1′)，7.37(m，5H，H-2′，3′，4′， 5′，6′)，2.55-2.72(s，each，3H，6×CH₃-pyrazine ring)，1.00-2.50(26H， methyl-and methylene-of steroid structure).13C-NMR(CDCl3)(ppm)：35.3 (C1)，26.5(C2)，75.6(C3)，26.4(C4)，45.6(C5)，72.9(C6)，31.3(C7)，34.8 (C8)，39.8(C9)，36.3(C10)，20.7(C11)，39.9(C12)，42.9(C13)，56.1(C14)， 24.2(C15)，28.1(C16)，55.9(C17)，12.0(C18)，23.3(C19)，35.1(C20)，18.3 (C21)，31.0(C22)，30.9(C23)，174.0(C24)，66.1(1′-CH2)，136.1(C1′)，128.3 (C2′，6′)，128.6(C3′，5′)，128.2(C4′)；pyrazine ring：165.7(2″-C＝O)，154.1 (C2″)，140.4(C3″)，149.3(C5″)，150.6(C6″)，165.6(2″′-C＝O)，154.1(C2″′)， 140.3(C3″′)，149.3(C5″′)，150.2(C6″)，22.6(6″-CH₃)，22.2(5″-CH₃)，21.7 (3″-CH₃)，22.5(6″′-CH₃)，22.1(5″′-CH₃)，21.6(3″′-CH₃).HRMS(ESI)m/z： 779.47357[M+H]⁺，calcd.for C₄₇H₆₂N₄O₆ 778.46694.

EXAMPLE 16 preparation of Compound 14

Weighing about 0.3mmol of compound 12, placing in 50ml round bottom flask, adding 30ml THF, dissolving the mixture, adding Pd/C40 mg, reducing with hydrogen, stirring at room temperature for 10 hr, TLC monitoring reaction material disappearance, stopping reaction, filtering to remove Pd/C, concentrating the reaction solution under reduced pressure, and collecting the residue with 3ml CHCl₃Redissolving, adding 2.2g of silica gel, evaporating to dryness under reduced pressure, mixing the sample, and eluting with petroleum ether and acetone at a ratio of 5: 1 to obtain a white solid of 0.147 g.m.p.: 39.1-39.9 deg.C, yield 90.6%. 1H-NMR (CDCl)₃)(ppm)：0.69(s，3H，H-18)，1.05(s，3H，H-19)，0.95(d，J＝6.5Hz， 3H，H-21)，3.64(m，1H，H-3)，5.48(m，1H，H-6)，2.72(s，3H，CH₃-6″)，2.57 (brs，6H，CH₃-3″，5″)，1.00-2.50(28H，methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：35.4(C1)，30.1(C2)，71.3(C3)，28.1 (C4)，45.5(C5)，73.4(C6)，31.3(C7)，34.8(C8)，39.9(C9)，36.2(C10)，20.7 (C11)，39.9(C12)，43.0(C13)，56.0(C14)，24.2(C15)，30.2(C16)，56.0 (C17)，12.1(C18)，23.3(C19)，35.4(C20)，18.3(C21)，31.0(C22)，30.8 (C23)，178.8(C24)；pyrazine ring：165.6(2′-C＝O)，154.1(C2′)，140.3(C3′)， 149.3(C5′)，150.4(C6′)，22.5(6′-CH₃)，22.1(5′-CH₃)，21.5(3′-CH₃).HRMS (ESI)m/z：541.36359[M+H]⁺，calcd.for C₃₂H₄₈N₂O₅ 540.35632.

EXAMPLE 17 preparation of Compound 15

Weighing compound 13 about 0.3mmol, placing in 50ml round bottom flask, adding THF 30ml, dissolving the mixture, adding Pd/C40 mg, reducing with hydrogen, stirring at room temperature for 10 hr, TLC monitoring reaction raw material disappearance, stopping reaction, filtering to remove Pd/C, concentrating the reaction solution under reduced pressure, and collecting residue with 3ml CHCl₃Redissolving, adding 2.2g of silica gel, evaporating to dryness under reduced pressure, mixing the sample, and eluting with petroleum ether and acetone at a ratio of 5: 1 to obtain 0.147g of white solid. m.p.: 88.3-89.0 ℃, yield 91.5%. 1H-NMR (CDCl)₃)(ppm)：0.70(s，3H，H-18)，1.09(s，3H，H-19)，0.95(d，J＝6.5Hz， 3H，H-21)，5.04(m，1H，H-3)，5.51(m，1H，H-6)，2.55-2.73(s，each，3H， 6×CH3-pyrazine ring)，1.00-2.50(27H，methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：35.3(C1)，26.4(C2)，75.6(C3)，26.4 (C4)，45.6(C5)，73.0(C6)，31.3(C7)，34.8(C8)，39.9(C9)，36.3(C10)，20.7 (C11)，40.0(C12)，43.0(C13)，56.2(C14)，24.2(C15)，28.1(C16)，56.0 (C17)，12.0(C18)，23.3(C19)，35.2(C20)，18.3(C21)，31.0(C22)，30.8 (C23)，178.7(C24)；pyrazine ring：165.6(2′-C＝O)，154.1(C2′)，140.3(C3′)， 149.3(C5′)，150.6(C6′)，165.5(2″-C＝O)，154.1(C2″)，140.3(C3″)，149.3 (C5″)，150.3(C6′)，22.5(6′-CH₃)，22.1(5′-CH₃)，21.5(3′-CH₃)，22.4(6″-CH₃)， 22.0(5″-CH₃)，21.5(3″-CH₃).HRMS(ESI)m/z：689.42712[M+H]⁺，calcd. for C₄₀H₅₆N₄O₆ 688.41999.

EXAMPLE 18 preparation of Compound 16

Weighing 2.19mmol of 2-bromomethyl-3, 5, 6-trimethyl pyrazine and ursodesoxyPlacing cholic acid 2.19mmol in 50ml round bottom flask, adding 25ml DMF, dissolving the mixture, adding 3mmol potassium carbonate, stirring at 85 deg.C for 2 hr, TLC monitoring reaction raw material disappearance, stopping reaction, dispersing the reaction solution with large amount of saturated NaCl solution, extracting with 300ml ethyl acetate twice, separating ethyl acetate layer with anhydrous Na₂SO₄Drying, concentrating under reduced pressure, dissolving the residue with 3ml chloroform, adding 2.8g silica gel, evaporating under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 8: 1 to obtain white solid 0.596 g. m.p.: 46.2-46.9 ℃, yield 51.7%. 1H-NMR (CDCl)₃)(ppm)：0.65(s，3H， H-18)，0.94(s，3H，H-19)，0.91(d，J＝6.0Hz，3H，H-21)，3.58(brs，2H，H-3， 7)，5.19(t，J＝12.5Hz，2H，CH₂-2′)，2.52(s，3H，CH₃-6′)，2.51(s，3H， CH₃-5′)，2.50(s，3H，CH₃-3′)，1.00-2.50(28H，methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：35.0(C1)，30.3(C2)，71.33 (C3)，37.3(C4)，42.5(C5)，36.9(C6)，71.29(C7)，43.7(C8)，39.2(C9)，34.1 (C10)，21.2(C11)，40.1(C12)，43.8(C13)，55.8(C14)，26.9(C15)，28.6 (C16)，54.9(C17)，12.1(C18)，23.4(C19)，35.2(C20)，18.4(C21)，31.1 (C22)，31.0(C23)，173.8(C24)；pyrazine ring：64.9(2′-CH₂)，151.2(C2′)， 144.9(C3′)，149.0(C5′)，149.0(C6′)，21.6(6′-CH₃)，21.4(5′-CH₃)，20.4 (3′-CH₃).HRMS(ESI)m/z：527.38373[M+H]⁺，calcd.for C₃₂H₅₀N₂O₄ 526.37706.

EXAMPLE 19 preparation of Compound 17

Weighing about 1.2mmol of compound 16 and 1.8mmol of ligustrazine acid, placing in a 50ml round bottom flask, adding 25ml of CH₂Cl₂After the mixture is dissolved, respectively adding EDCI 1.8mmol and DMAP 0.18mmol, stirring at room temperature for 12h, monitoring the reaction raw material by TLC to basically disappear, stopping the reaction, concentrating under reduced pressure,adding 2.2g of silica gel, evaporating to dryness under reduced pressure, mixing the mixture with a sample, and eluting with petroleum ether and acetone at a ratio of 8: 1 to obtain 0.231g of white solid respectively. White Amorphous solid, m.p.: 59.1-59.9 deg.C, yield 28.6%. 1H-NMR (CDCl)₃)(ppm)：0.68(s，3H，H-18)，1.00(s，3H，H-19)，0.92 (d，J＝6.0Hz，3H，H-21)，5.00(m，1H，H-3)，3.60(m，1H，H-7)，5.20(t，J＝ 12.5Hz，2H，CH₂-2′)，2.72(s，3H，CH₃-6″)，2.58(s，3H，CH₃-5″)，2.57(s，3H， CH₃-3″)，2.54(s，3H，CH₃-6′)，2.52(s，3H，CH₃-5′)，2.51(s，3H，CH₃-3′)， 1.00-2.50(27H，methyl-and methylene-of steroid structure).13C-NMR (CDCl₃)(ppm)：34.7(C1)，26.5(C2)，75.7(C3)，33.1(C4)，42.4(C5)，36.7 (C6)，71.2(C7)，43.7(C8)，39.1(C9)，34.2(C10)，21.5(C11)，40.1(C12)， 43.7(C13)，55.7(C14)，26.9(C15)，28.6(C16)，54.9(C17)，12.1(C18)，23.3 (C19)，35.2(C20)，18.4(C21)，31.1(C22)，31.0(C23)，173.9(C24)；pyrazine ring：65.0(2′-CH2)，151.2(C2′)，144.9(C3′)，149.0(C5′)，149.0(C6′)，165.7 (2″-C＝O)，154.1(C2″)，140.3(C3″)，149.3(C5″)，150.6(C6″)，21.5(6′-CH₃)， 21.2(5′-CH₃)，20.5(3′-CH₃)，22.6(6″-CH₃)，22.1(5″-CH₃)，21.7(3″-CH₃). HRMS(ESI)m/z：675.44788[M+H]⁺，calcd.for C₄₀H₅₈N₄O₅674.44072.

EXAMPLE 20 preparation of Compound 18

This structure was isolated in the preparation of compound 17 as a colorless oil, 0.155 g. yield 15.7%. 1H-NMR (CDCl)₃)(ppm)：0.68(s，3H，H-18)，1.01(s，3H，H-19)，0.91 (d，J＝6.5Hz，3H，H-21)，5.02(m，1H，H-3)，5.11(m，1H，H-7)，5.17(t，J＝ 12.5Hz，2H，CH₂-2′)，2.48-2.71(s，each，3H，9×CH₃-pyrazine ring)， 1.00-2.50(26H，methyl-and methylene-of steroid structure).13C-NMR (CDCl₃)(ppm)：34.6(C1)，26.4(C2)，75.5(C3)，32.8(C4)，40.0(C5)，32.9 (C6)，75.4(C7)，42.4(C8)，39.6(C9)，34.2(C10)，21.6(C11)，39.9(C12)， 43.6(C13)，55.3(C14)，25.9(C15)，28.4(C16)，55.0(C17)，12.1(C18)，23.3 (C19)，35.2(C20)，18.3(C21)，31.0(C22)，30.9(C23)，173.8(C24)；pyrazine ring：64.9(2′-CH₂)，151.2(C2′)，144.9(C3′)，148.9(C5′)，149.0(C6′)，165.7 (2″-C＝O)，154.1(C2″)，140.2(C3″)，149.3(C5″)，150.6(C6″)，165.7 (2″′-C＝O)，153.9(C2″′)，140.2(C3″′)，149.2(C5″′)，150.7(C6″)，21.4 (6′-CH₃)，21.3(5′-CH₃)，20.5(3′-CH₃)，22.6(6″-CH₃)，22.1(5″-CH₃)，21.7 (3″-CH₃)，22.6(6″′-CH₃)，22.2(5″′-CH₃)，21.6(3″′-CH₃).HRMS(ESI)m/z： 823.51196[M+H]⁺，calcd.for C₄₈H₆₆N₆O₆ 822.50438.

Preparation of ursodeoxycholic acid benzyl ester: weighing benzyl bromide 5.0mmol and ursodeoxycholic acid 5.0mmol, placing in 50ml round bottom flask, adding DMF 25ml, dissolving the mixture, adding potassium carbonate 5mmol, stirring at 85 deg.C for 2 hr, monitoring by TLC that the reaction raw materials basically disappear, stopping reaction, adding a large amount of saturated NaCl solution into the reaction solution for dispersion, extracting with ethyl acetate 400ml twice, and using anhydrous Na for ethyl acetate layer₂SO₄Drying, concentrating under reduced pressure, dissolving the residue with 5ml chloroform, adding 3.8g silica gel, evaporating under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 5: 1 to obtain 2.181g white solid. White powder, m.p.: 142.5-143.2 ℃ and yield 90.5 percent.

EXAMPLE 21 preparation of Compound 19

Weighing 3.0mmol of ursodeoxycholic acid benzyl ester and 4.0mmol of ligustrazine acid, placing in a 50ml round bottom flask, adding 30ml of CH₂Cl₂After the mixture is dissolved, respectively adding EDCI 4.0mmol and DMAP 0.4mmol, stirring at room temperature for 12h, monitoring by TLC that the reaction raw material basically disappears, stopping the reaction, washing the reaction solution twice by 60ml of saturated NaCl solution, and obtaining CH₂Cl₂Anhydrous Na for layer₂SO₄Drying, concentrating under reduced pressure, adding 3.2g silica gel, evaporating to dryness under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 7: 1 to obtain white solid 0.486 g. White Amorphous solid, m.p.: 58.4-59.2 deg.C, yield 25.7%. 1H-NMR (CDCl)₃)(ppm)：0.68(s，3H，H-18)，1.00(s，3H，H-19)，0.93(d，J＝ 6.5Hz，3H，H-21)，5.00(m，1H，H-3)，3.61(m，1H，H-7)，5.14，5.11(ea，d，J ＝12.0Hz，1H，CH2-1′)，7.37(m，5H，H-2′，3′，4′，5′，6′)，2.73(s，3H，CH₃-6″)， 2.58(s，3H，CH₃-5″)，2.57(s，3H，CH₃-3″)，1.00-2.50(27H，methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：34.7(C1)，26.5 (C2)，75.7(C3)，33.1(C4)，42.5(C5)，36.7(C6)，71.2(C7)，43.8(C8)，39.2 (C9)，34.2(C10)，21.2(C11)，40.1(C12)，43.8(C13)，55.7(C14)，26.9(C15)， 28.6(C16)，55.0(C17)，12.1(C18)，23.3(C19)，35.2(C20)，18.4(C21)，31.3 (C22)，31.0(C23)，174.0(C24)，66.1(1′-CH2)，136.1(C1′)，128.2(C2′，6′)， 128.5(C3′，5′)，128.2(C4′)；pyrazine ring：165.7(2″-C＝O)，154.1(C2″)， 140.3(C3″)，149.2(C5″)，150.5(C6″)，22.5(6″-CH₃)，22.1(5″-CH₃)，21.6 (3″-CH₃).HRMS(ESI)m/z：631.41046[M+H]⁺，calcd.for C₃₉H₅₄N₂O₅630.40327.

EXAMPLE 22 preparation of Compound 20

This structure was isolated in the preparation of compound 19, 0.334 g. White solid, m.p.: 147.2-148.0 deg.C, yield 14.3%. 1H-NMR (CDCl)₃)(ppm)：0.68(s，3H，H-18)， 1.07(s，3H，H-19)，0.91(d，J＝6.0Hz，3H，H-21)，5.02(m，2H，H-3，7)，5.10 (brs，2H，CH₂-1′)，7.37(m，5H，H-2′，3′，4′，5′，6′)，2.56-2.75(s，each，3H， 6×CH₃-pyrazine ring)，1.00-2.50(26H，methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：34.7(C1)，26.5(C2)，75.5(C3)，32.9 (C4)，40.1(C5)，32.9(C6)，75.3(C7)，42.4(C8)，39.7(C9)，34.2(C10)，21.3 (C11)，40.0(C12)，43.7(C13)，55.4(C14)，25.9(C15)，28.3(C16)，55.1 (C17)，12.1(C18)，23.3(C19)，35.1(C20)，18.4(C21)，31.3(C22)，31.0 (C23)，173.9(C24)，66.0(1′-CH2)，136.2(C1′)，128.1(C2′，6′)，128.5(C3′，5′)， 128.1(C4′)；pyrazine ring：165.7(2″-C＝O)，153.9(C2″)，140.3(C3″)，149.1 (C5″)，150.5(C6″)，165.7(2″′-C＝O)，153.7(C2″′)，140.3(C3″′)，149.1(C5″′)， 150.5(C6″)，22.4(6″-CH₃)，22.0(5″-CH₃)，21.5(3″-CH₃)，22.4(6″′-CH₃)， 22.0(5″′-CH₃)，21.5(3″′-CH₃).HRMS(ESI)m/z：779.47418[M+H]⁺，calcd. for C₄₇H₆₂N₄O₆ 778.46694.

EXAMPLE 23 preparation of Compound 21

Weighing about 0.6mmol of compound 19, placing in a 50ml round-bottom flask, adding 30ml THF, adding Pd (OH) after the mixture is dissolved₂40mg of/C, hydrogen reduction, stirring at room temperature for 10 hours, monitoring by TLC that the reaction material is substantially disappeared, stopping the reaction, filtering to remove Pd (OH)₂C, concentrating the reaction solution under reduced pressure, and adding 3ml of CHCl into the residue₃Redissolving, adding 2.2g of silica gel, evaporating to dryness under reduced pressure, mixing the sample, and eluting with petroleum ether and acetone at a ratio of 5: 1 to obtain 0.301g of white solid. White Amorphous solid, m.p.: 101.9-102.7 ℃, yield 92.8%. 1H-NMR (CDCl)₃)(ppm)：0.71(s，3H，H-18)， 1.01(s，3H，H-19)，0.96(d，J＝6.5Hz，3H，H-21)，5.01(m，1H，H-3)，3.62(m， 1H，H-7)，2.73(s，3H，CH₃-6′)，2.58(s，3H，CH₃-5′)，2.58(s，3H，CH₃-3′)， 1.00-2.50(28H，methyl-and methylene-of steroid structure).13C-NMR (CDCl₃)(ppm)：34.7(C1)，26.5(C2)，75.7(C3)，33.1(C4)，42.4(C5)，36.7 (C6)，71.3(C7)，43.7(C8)，39.2(C9)，34.2(C10)，21.2(C11)，40.1(C12)， 43.8(C13)，55.7(C14)，26.9(C15)，28.6(C16)，55.0(C17)，12.1(C18)，23.3 (C19)，35.2(C20)，18.4(C21)，31.0(C22)，30.8(C23)，178.9(C24)；pyrazine ring：165.6(2′-C＝O)，154.2(C2′)，140.3(C3′)，149.3(C5″)，150.6(C6′)，22.5 (6′-CH₃)，22.1(5′-CH₃)，21.6(3′-CH₃).HRMS(ESI)m/z：541.36322[M+H]⁺， calcd.for C₃₂H₄₈N₂O₅ 540.35632.

EXAMPLE 24 preparation of Compound 22

Weighing about 0.3mmol of compound 20, placing in a 50ml round bottom flask, adding 30ml THF, adding Pd (OH) after the mixture is dissolved₂25 mg/C, hydrogen reduction, stirring at room temperature for 10h, monitoring by TLC that the reaction material is substantially disappeared, stopping the reaction, filtering to remove Pd (OH)₂C, concentrating the reaction solution under reduced pressure, and adding 3ml of CHCl into the residue₃Redissolving, adding 2.2g silica gel, drying by distillation under reduced pressure, mixing the sample, and eluting with petroleum ether and acetone at a ratio of 5: 1 to obtain 0.193g white solid. m.p.: 147.2-148.0 deg.C, yield 93.4%. 1H-NMR (CDCl)₃)(ppm)：0.71(s，3H，H-18)，1.07(s，3H，H-19)，0.93 (d，J＝6.0Hz，3H，H-21)，5.02(m，1H，H-3)，5.12(m，1H，H-7)，2.57-2.80(s， each，3H，6×CH₃-pyrazine ring)，1.00-2.50(27H，methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：34.6(C1)，26.4(C2)，75.5(C3)， 32.8(C4)，40.0(C5)，32.9(C6)，75.4(C7)，42.4(C8)，39.6(C9)，34.2(C10)， 21.3(C11)，39.9(C12)，43.7(C13)，55.4(C14)，25.9(C15)，28.4(C16)，55.0 (C17)，12.1(C18)，23.3(C19)，35.2(C20)，18.3(C21)，30.8(C22)，30.9 (C23)，178.6(C24)；pyrazine ring：165.6(2′-C＝O)，154.1(C2′)，140.2(C3′)， 149.3(C5′)，150.6(C6′)，165.6(2″-C＝O)，153.9(C2″)，140.2(C3″)，149.2 (C5″)，150.6(C6′)，22.5(6′-CH₃)，22.0(5′-CH₃)，21.5(3′-CH₃)，22.4(6″-CH3)， 22.0(5″-CH₃)，21.5(3″-CH₃).HRMS(ESI)m/z：689.42834[M+H]⁺，calcd. for C₄₀H₅₆N₄O₆ 688.41999.

EXAMPLE 25 preparation of Compound 23

Weighing 2.19mmol of 2-bromomethyl-3, 5, 6-trimethylpyrazine and 2.19mmol of chenodeoxycholic acid, placing the materials in a 50ml round-bottom flask, adding 25ml of DMF, adding 3mmol of potassium carbonate after the mixture is dissolved, stirring for 2h at 85 ℃, monitoring the basic disappearance of reaction raw materials by TLC, stopping the reaction, adding a large amount of saturated NaCl solution into the reaction solution for dispersion, extracting twice by 300ml of ethyl acetate, and using anhydrous Na for an ethyl acetate layer₂SO₄Drying, concentrating under reduced pressure, dissolving the residue with 3ml chloroform, adding 2.8g silica gel, evaporating under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 8: 1 to obtain white solid 0.638 g. m.p.: 64.8-64.6 ℃, yield 55.4%. 1H-NMR (CDCl)₃)(ppm)：0.64(s，3H， H-18)，0.90(s，3H，H-19)，0.91(d，J＝6.5Hz，3H，H-21)，3.44(m，1H，H-3)， 3.84(brs，1H，H-7)，5.19(t，J＝12.5Hz，2H，CH₂-2′)，2.53(s，3H，CH₃-6′)， 2.51(s，3H，CH₃-5′)，2.50(s，3H，CH₃-3′)，1.00-2.50(28H，methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：35.3(C1)，30.7 (C2)，71.9(C3)，39.8(C4)，41.5(C5)，34.7(C6)，68.3(C7)，39.6(C8)，32.8 (C9)，35.0(C10)，20.6(C11)，39.4(C12)，42.6(C13)，50.4(C14)，23.6(C15)， 28.1(C16)，55.8(C17)，11.7(C18)，22.8(C19)，35.4(C20)，18.2(C21)，31.0 (C22)，31.0(C23)，173.9(C24)；pyrazine ring：64.9(2′-CH₂)，151.2(C2′)， 144.9(C3′)，148.9(C5′)，149.0(C6′)，21.6(6′-CH₃)，21.4(5′-CH₃)，20.5 (3′-CH₃).HRMS(ESI)m/z：527.38440[M+H]+，calcd.for C₃₂H₅₀N₂O₄526.37706.

EXAMPLE 26 preparation of Compound 24

Weighing about 1.2mmol of compound 23 and 1.8mmol of ligustrazine acid, placing in a 50ml round bottom flask, adding 25ml of CH₂Cl₂After the mixture is dissolved, EDCI 1.8mmol and DMAP 0.18mmol are respectively added, the mixture is stirred for 12h at room temperature, TLC monitors that the reaction raw materials basically disappear, the reaction is stopped, the mixture is concentrated under reduced pressure, 2.2g of silica gel is added, the mixture is evaporated to dryness under reduced pressure and is mixed, and eluent is petroleum ether and acetone which are 7: 1 to elute, so that 0.373g of white oily matter is obtained. Oil, yield 46.1%. 1H-NMR (CDCl)₃)(ppm)：0.66(s， 3H，H-18)，0.95(s，3H，H-19)，0.92(d，J＝6.0Hz，3H，H-21)，4.91(m，1H， H-3)，3.87(brs，1H，H-7)，5.19(t，J＝12.5Hz，2H，CH₂-2′)，2.71(s，3H， CH₃-6″)，2.56(s，3H，CH₃-5″)，2.55(s，3H，CH₃-3″)，2.53(s，3H，CH₃-6′)，2.51(s，3H，CH3-5′)，2.51(s，3H，CH3-3′)，1.00-2.50(27H，methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：35.2(C1)，26.7 (C2)，76.3(C3)，35.4(C4)，41.4(C5)，34.5(C6)，68.4(C7)，39.6(C8)，32.8 (C9)，35.1(C10)，20.6(C11)，39.4(C12)，42.7(C13)，50.4(C14)，23.7(C15)， 28.1(C16)，55.8(C17)，11.8(C18)，22.7(C19)，35.3(C20)，18.2(C21)，31.0 (C22)，31.0(C23)，173.9(C24)；pyrazine ring：64.9(2′-CH₂)，151.2(C2′)， 144.9(C3′)，149.0(C5′)，149.0(C6′)，165.8(2″-C＝O)，153.9(C2″)，140.5 (C3″)，149.2(C5″)，150.4(C6″)，21.5(6′-CH₃)，21.4(5′-CH₃)，20.5(3′-CH₃)，22.6(6″-CH₃)，22.1(5″-CH₃)，21.6(3″-CH₃).HRMS(ESI)m/z：675.44879 [M+H]⁺，calcd.for C₄₀H₅₈N₄O₅ 674.44072.

Preparing chenodeoxycholic acid benzyl ester: weighing benzyl bromide 5.0mmol and chenodeoxycholic acid 5.0mmol, placing in 50ml round bottom flask, adding 25ml DMF, dissolving the mixture, adding 5mmol potassium carbonate, stirring at 85 deg.C for 2h, monitoring by TLC that the reaction raw materials basically disappear, stopping the reaction, adding a large amount of saturated NaCl solution into the reaction solution for dispersion, extracting twice with 400ml ethyl acetate, using anhydrous Na for ethyl acetate layer₂SO₄Drying, concentrating under reduced pressure, dissolving the residue with 5ml chloroform, adding 3.8g silica gel, evaporating under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 5: 1 to obtain white solid 2.157 g. White powder, m.p.: 146.9-147.7 ℃ and yield 89.5 percent.

EXAMPLE 27 preparation of Compound 25

3.0mmol of chenodeoxycholic acid benzyl ester and 4.0mmol of ligustrazine acid are weighed and placed in a 50ml round-bottom flask, and 30ml of CH is added₂Cl₂After the mixture is dissolved, respectively adding EDCI 4.0mmol and DMAP 0.4mmol, stirring at room temperature for 12h, monitoring by TLC that the reaction raw material basically disappears, stopping the reaction, washing the reaction solution twice by 60ml of saturated NaCl solution, and obtaining CH₂Cl₂Anhydrous Na for layer2SO4, concentrating under reduced pressure, adding 3.2g silica gel, evaporating to dryness under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 7: 1 to obtain white solid 0.911 g. m.p.: 64.8-64.6 ℃, yield 48.2%. 1H-NMR (CDCl)₃) (ppm)：0.65(s，3H，H-18)，0.96(s，3H，H-19)，0.92(d，J＝6.0Hz，3H，H-21)， 4.92(m，1H，H-3)，3.87(brs，1H，H-7)，5.14，5.10(ea，d，J＝12.5Hz，1H， CH2-1′)，7.36(m，5H，H-2′，3′，4′，5′，6′)，2.72(s，3H，CH₃-6″)，2.56(s，3H，CH₃-5″)，2.56(s，3H，CH₃-3″)，1.00-2.50(27H，methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：35.2(C1)，26.8(C2)，76.3(C3)， 35.2(C4)，41.4(C5)，34.5(C6)，68.4(C7)，39.6(C8)，32.8(C9)，35.1(C10)， 20.6(C11)，39.4(C12)，42.7(C13)，50.4(C14)，23.7(C15)，28.1(C16)，55.8 (C17)，11.8(C18)，22.7(C19)，35.3(C20)，18.3(C21)，31.3(C22)，31.0 (C23)，174.0(C24)，66.1(1′-CH₂)，136.1(C1′)，128.2(C2′，6′)，128.5(C3′，5′)， 128.1(C4′)；pyrazine ring：165.8(2″-C＝O)，153.8(C2″)，140.6(C3″)，149.2 (C5″)，150.3(C6″)，22.5(6″-CH₃)，22.1(5″-CH₃)，21.5(3″-CH₃).HRMS(ESI) m/z：631.40985[M+H]⁺，calcd.for C₃₉H₅₄N₂O₅ 630.40327.

EXAMPLE 28 preparation of Compound 26

Weighing compound 25 about 1.2mmol, placing in 50ml round bottom flask, adding THF 30ml, dissolving the mixture, adding Pd/C78 mg, reducing with hydrogen, stirring at room temperature for 10 hr, TLC monitoring reaction material disappearance, stopping reaction, filtering to remove Pd/C, concentrating the reaction solution under reduced pressure, and collecting residue with 3ml CHCl₃Redissolving, adding 2.2g of silica gel, evaporating to dryness under reduced pressure, mixing the sample, and eluting with petroleum ether and acetone at a ratio of 5: 1 to obtain 0.588g of white solid. m.p.: 130.0-130.8 ℃, yield 90.7%. 1H-NMR (C)DCl₃)(ppm)：0.68(s，3H，H-18)，0.96(s，3H，H-19)，0.95(d，J＝ 7.5Hz，3H，H-21)，4.92(m，1H，H-3)，3.87(brs，1H，H-7)，2.72(s，3H， CH₃-6′)，2.65(s，6H，CH₃-3′，5′)，1.00-2.50(28H，methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：35.2(C1)，26.7(C2)，76.3(C3)， 35.2(C4)，41.4(C5)，34.5(C6)，68.5(C7)，39.6(C8)，32.8(C9)，35.1(C10)， 20.6(C11)，39.4(C12)，42.7(C13)，50.4(C14)，23.7(C15)，28.1(C16)，55.8 (C17)，11.8(C18)，22.7(C19)，35.4(C20)，18.3(C21)，31.0(C22)，30.8 (C23)，179.1(C24)；pyrazine ring：165.7(2′-C＝O)，153.9(C2′)，140.5(C3′)，149.3(C5′)，150.4(C6′)，22.5(6′-CH₃)，22.0(5′-CH₃)，21.4(3′-CH₃).HRMS (ESI)m/z：541.36346[M+H]⁺，calcd.for C₃₂H₄₈N₂O₅ 540.35632.

EXAMPLE 29 preparation of Compound 27

Weighing 2.19mmol of 2-bromomethyl-3, 5, 6-trimethyl pyrazine and 2.19mmol of lithocholic acid, placing in a 50ml round bottom flask, adding 25ml DMF, dissolving the mixture, adding 3mmol of potassium carbonate, stirring at 85 deg.C for 2h, monitoring by TLC that the reaction raw materials basically disappear, stopping the reaction, adding a large amount of saturated NaCl solution into the reaction solution for dispersion, extracting twice with 300ml ethyl acetate, and extracting the ethyl acetate layer with anhydrous Na₂SO₄Drying, concentrating under reduced pressure, dissolving the residue with 3ml chloroform, adding 2.8g silica gel, evaporating under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 6: 1 to obtain white solid 0.589 g. m.p.: 119.3-119.9 deg.C, yield 52.7%. 1H-NMR (CDCl)₃)(ppm)：0.60(s，3H，H-18)， 0.89(s，3H，H-19)，0.88(d，J＝6.0Hz，3H，H-21)，3.60(m，H，H-3)，5.17(t，J ＝12.5Hz，2H，CH₂-2′)，2.51(s，3H，CH₃-6′)，2.49(s，3H，CH₃-5′)，2.48(s，3H， CH₃-3′)，1.00-2.50(29H，methyl-and methylene-of steroid structure). 13C-NMR(CDCl₃)(ppm)：35.8(C1)，30.5(C2)，71.7(C3)，36.4(C4)，42.1 (C5)，27.2(C6)，26.4(C7)，40.2(C8)，34.6(C9)，35.3(C10)，20.8(C11)，40.4 (C12)，42.7(C13)，56.5(C14)，24.2(C15)，28.1(C16)，56.0(C17)，12.0 (C18)，23.4(C19)，35.4(C20)，18.2(C21)，31.1(C22)，31.0(C23)，173.8 (C24)；pyrazine ring：64.9(2′-CH₂)，151.2(C2′)，144.9(C3′)，148.9(C5′)， 149.0(C6′)，21.6(6′-CH₃)，21.4(5′-CH₃)，20.4(3′-CH₃).HRMS(ESI)m/z： 511.38754[M+H]⁺，calcd.for C₃₂H₅₀N₂O₃ 510.38214.

EXAMPLE 30 preparation of Compound 28

Weighing about 1.2mmol of compound 27 and 1.2mmol of ligustrazine acid, placing in a 50ml round bottom flask, adding 25ml of CH₂Cl₂After the mixture is dissolved, EDCI 1.2mmol and DMAP 0.12mmol are respectively added, the mixture is stirred for 12h at room temperature, TLC monitors that the reaction raw materials basically disappear, the reaction is stopped, the mixture is concentrated under reduced pressure, 2.2g of silica gel is added, the mixture is evaporated to dryness under reduced pressure and mixed, and the eluent is petroleum ether and acetone which are 5: 1 for elution, so that 0.386g of oily matter is respectively obtained. m.p.: 85.3-85.9 deg.C, yield 48.9%. 1H-NMR (CDCl)₃) (ppm)：0.65(s，3H，H-18)，0.97(s，3H，H-19)，0.91(d，J＝6.0Hz，3H，H-21)， 5.01(m，H，H-3)，5.20(t，J＝12.5Hz，2H，CH₂-2′)，2.73(s，3H，CH₃-6″)，2.58 (s，3H，CH₃-5″)，2.57(s，3H，CH₃-3″)，2.54(s，3H，CH₃-6′)，2.53(s，3H， CH₃-5′)，2.52(s，3H，CH₃-3′)，1.00-2.50(28H，methyl-and methylene-of steroid structure).13C-NMR(CDCl₃)(ppm)：35.8(C1)，27.1(C2)，76.3(C3)， 32.2(C4)，42.1(C5)，26.6(C6)，26.3(C7)，40.1(C8)，34.7(C9)，35.1(C10)， 20.8(C11)，40.4(C12)，42.7(C13)，56.5(C14)，24.2(C15)，28.2(C16)，56.0 (C17)，12.0(C18)，23.3(C19)，35.3(C20)，18.3(C21)，31.1(C22)，31.0 (C23)，173.9(C24)；pyrazine ring：65.0(2′-CH2)，151.2(C2′)，144.9(C3′)， 149.0(C5′)，149.0(C6′)，165.5(2″-C＝O)，154.0(C2″)，140.5(C3″)，149.2 (C5″)，150.4(C6″)，21.7(6′-CH₃)，21.5(5′-CH₃)，20.5(3′-CH₃)，22.6(6″-CH₃)， 22.1(5″-CH₃)，21.7(3″-CH₃).HRMS(ESI)m/z：659.45245[M+H]⁺，calcd. for C₄₀H₅₈N₄O₄ 658.44581.

Preparation of lithocholic acid benzyl ester: weighing benzyl bromide 5.0mmol and lithocholic acid 5.0mmol, placing in 50ml round bottom flask, adding 25ml DMF, dissolving the mixture, adding 2.0mmol potassium carbonate, adding 5mmol potassium carbonate, stirring at 85 deg.C for 2 hr, monitoring by TLC that the reaction raw material basically disappears, stopping reaction, adding a large amount of saturated NaCl solution to the reaction solution for dispersion, extracting with 400ml ethyl acetate twice, extracting the ethyl acetate layer with anhydrous Na₂SO₄Drying, concentrating under reduced pressure, dissolving the residue with 5ml chloroform, adding 3.8g silica gel, evaporating under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 5: 1 to obtain white solid 2.125 g. White powder, m.p.: 129.1-129.8 ℃ and yield 91.2 percent.

EXAMPLE 31 preparation of Compound 29

Weighing lithocholic acid benzyl ester 3.0mmol and ligustrazine acid 3.0mmol, placing in 50ml round bottom flask, adding 30ml CH₂Cl₂After the mixture is dissolved, respectively adding EDCI 3.0mmol and DMAP 0.3mmol, stirring at room temperature for 12h, monitoring by TLC that the reaction raw material basically disappears, stopping the reaction, washing the reaction solution twice by 60ml of saturated NaCl solution, and obtaining CH₂Cl₂Anhydrous Na for layer₂SO₄Drying, concentrating under reduced pressure, adding 3.2g silica gel, evaporating to dryness under reduced pressure, mixing, eluting with petroleum ether and acetone at ratio of 7: 1 to obtain white solid 0.938 g. m.p.: 102.6-103.3 deg.C, yield 50.9%. 1H-NMR (CDCl)₃) (ppm)：0.65(s，3H，H-18)，0.98(s，3H，H-19)，0.92(d，J＝6.5Hz，3H，H-21)， 5.06(m，H，H-3)，5.15，5.12(ea，d，J＝12.5Hz，1H，CH₂-1′)，7.38(m，5H， H-2′，3′，4′，5′，6′)，2.74(s，3H，CH₃-6″)，2.59(s，3H，CH₃-5″)，2.58(s，3H， CH₃-3″)，1.00-2.50(28H，methyl-and methylene-of steroid structure). 13C-NMR(CDCl₃)(ppm)：35.8(C1)，27.1(C2)，76.3(C3)，32.3(C4)，42.1 (C5)，26.6(C6)，26.4(C7)，40.1(C8)，34.7(C9)，35.1(C10)，20.9(C11)，40.4 (C12)，42.7(C13)，56.5(C14)，24.2(C15)，28.2(C16)，56.0(C17)，12.0 (C18)，23.3(C19)，35.3(C20)，18.3(C21)，31.3(C22)，31.0(C23)，174.1 (C24)，66.1(1′-CH2)，136.1(C1′)，128.2(C2′，6′)，128.5(C3′，5′)，128.2(C4′)；pyrazine ring：165.8(2″-C＝O)，154.0(C2″)，140.6(C3″)，149.3(C5″)，150.4 (C6″)，22.6(6″-CH₃)，22.1(5″-CH₃)，21.7(3″-CH₃).HRMS(ESI)m/z： 637.39772[M+Na]⁺，calcd.for C₃₉H₅₄N₂O₄ 614.40836.

EXAMPLE 32 preparation of Compound 30

Weighing 291.2mmol of compound, placing in 50mL round-bottom flask, adding 30mL THF, dissolving the mixture, adding 40mg Pd/C, reducing with hydrogen, stirring at room temperature for 10h, monitoring by TLC that the reaction material is substantially disappeared, stopping the reaction, filtering to remove Pd/C, concentrating the reaction solution under reduced pressure, and adding 3ml CHCl into the residue₃Redissolving, adding 2.2g of silica gel, drying by distillation under reduced pressure, mixing the sample, and eluting by petroleum ether and acetone of 5: 1 to obtain 0.583g of white solid. m.p.: 184.5-185.2 deg.C，yield 92.7％.¹H-NMR (CDCl₃)(ppm)：0.68(s，3H，H-18)，0.99(s，3H，H-19)，0.95(d，J＝6.5Hz， 3H，H-21)，5.07(m，H，H-3)，2.74(s，3H，CH ₃-6″)，2.59(s，3H，CH ₃-5″)，2.59 (s，3H，CH ₃-3″)，1.00-2.50(29H，methyl-and methylene-of steroid structure). ¹³C-NMR(CDCl₃)(ppm)：35.8(C1)，27.1(C2)，76.3(C3)，32.3(C4)，42.1 (C5)，26.6(C6)，26.4(C7)，40.1(C8)，34.7(C9)，35.1(C10)，20.9(C11)，40.4 (C12)，42.7(C13)，56.5(C14)，24.2(C15)，28.2(C16)，56.0(C17)，12.0 (C18)，23.3(C19)，35.3(C20)，18.3(C21)，31.3(C22)，31.0(C23)，174.1 (C24)，66.1(1′-CH₂)，136.1(C1′)，128.2(C2′，6′)，128.5(C3′，5′)，128.2(C4′)； pyrazine ring：165.8(2″-C＝O)，154.0(C2″)，140.6(C3″)，149.3(C5″)，150.4 (C6″)，22.6(6″-CH₃)，22.1(5″-CH₃)，21.7(3″-CH₃).HRMS(ESI)m/z： 525.36871[M+H]⁺，calcd.for C₃₂H₄₈N₂O₄524.36141.

Example 33

Taking 10g of the compound prepared in any one of the embodiments 3-32, adding appropriate auxiliary materials into injection (including freeze-dried powder injection and sterile subpackaged powder injection), and preparing the anti-tumor drug injection according to the injection (including freeze-dried powder injection and sterile subpackaged powder injection) process.

Example 34

Taking 10g of the compound prepared in any one of the embodiments 3-32, adding appropriate auxiliary materials into tablets (including sustained release tablets, matrix tablets, coated tablets, dispersible tablets and the like), and preparing the compound into the anti-tumor tablets according to the process of the tablets (including sustained release tablets, matrix tablets, coated tablets, dispersible tablets and the like).

Example 35

Taking 10g of the compound prepared in any one of the embodiments 3-32, adding appropriate capsule auxiliary materials, and preparing the compound into an anti-tumor capsule according to a capsule process.

Example 36

Taking 10g of the compound prepared in any one of the embodiments 3-32, adding appropriate auxiliary materials of an emulsion (including microemulsion, nanoemulsion and the like), and preparing the anti-tumor emulsion according to the process of the emulsion (including microemulsion, nanoemulsion and the like).

Example 37

Taking 10g of the compound prepared in any one of the embodiments 3-32, adding proper auxiliary materials of granules, and preparing the antitumor granules according to a granule process.

Example 38

Taking 10g of the compound prepared in any one of the embodiments 3-32, adding a proper auxiliary material of the sustained-release controlled-release agent, and preparing the sustained-release controlled-release agent of the antitumor drug according to the sustained-release controlled-release agent process.

Example 39

Taking 10g of the compound prepared in any one of the embodiments 3-32, adding proper auxiliary materials of the oral liquid, and preparing the anti-tumor oral liquid according to an oral liquid process.

Example 40

Taking 10g of the compound prepared in any one of the embodiments 3-32, adding proper auxiliary materials of a liposome formulation, and preparing the antitumor liposome formulation according to a liposome process.

Claims (8)

1. A compound having the structure of formula 1 or a pharmaceutically acceptable salt thereof:

2. a pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

3. A process for preparing a compound according to claim 1, comprising:

the hyodeoxycholic acid and 2-bromomethyl-3, 5, 6-trimethylpyrazine react under alkaline conditions, and products obtained by the reaction further react with ligustrazine acid under the action of a catalyst and a condensing agent.

4. Use of a compound according to claim 1 or a pharmaceutically acceptable salt thereof in the manufacture of an anti-cancer medicament.

5. The use of claim 4, wherein the cancer is liver cancer, colon cancer, cervical cancer, breast cancer.

6. A kit comprising a therapeutically effective amount of a compound of formula 1 according to claim 1 or a pharmaceutically acceptable salt thereof and an anti-cancer agent; wherein the therapeutically effective amount of the compound of formula 1 or a pharmaceutically acceptable salt thereof is 1-10 mg/kg-day.

7. The kit of claim 6, wherein said anticancer agent is any one or more of cyclophosphamide, 5-fluorouracil, paclitaxel, doxorubicin, etoposide, irinotecan, oxaliplatin, cisplatin, or gemcitabine.

8. Use of a kit according to claim 6 or 7 for the manufacture of an anti-cancer medicament.