CN110746479A

CN110746479A - Compound and preparation method and application thereof

Info

Publication number: CN110746479A
Application number: CN201910968512.XA
Authority: CN
Inventors: 刘玉法; 高修正; 吕绪涛; 高翔宇; 孙彬
Original assignee: Shandong Normal University
Current assignee: Shandong Normal University
Priority date: 2019-10-12
Filing date: 2019-10-12
Publication date: 2020-02-04
Anticipated expiration: 2039-10-12
Also published as: CN110746479B

Abstract

The invention provides a compound, a preparation method and application thereof, wherein the compound has a structure shown in a formula (I):

Description

Compound and preparation method and application thereof

Technical Field

The invention relates to the field of biological medicines, and particularly relates to a compound and a preparation method and application thereof.

Background

The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.

Cholangiocarcinoma is a common biliary tract malignant tumor, with high malignancy, very poor prognosis, and increasing morbidity and mortality year by year (wangxian, et al, the clinical treatment progress of hepatoportal cholangiocarcinoma [ J ], electronic journal of Chinese liver surgery, 2018, 7(4): 253-containing 257). The common chemotherapy drugs in the clinic of bile duct cancer are 5-fluorouracil and cisplatin. When the 5-fluorouracil reaches 20ug/mL, the traditional Chinese medicine composition has a good effect of inhibiting the growth of human bile duct cancer QBC939 cells, and has side effects of bone marrow inhibition such as leukopenia and platelet drop, and gastrointestinal reactions such as inappetence, nausea, vomiting, stomatitis, gastritis, abdominal pain and diarrhea. When the cisplatin reaches 3.0 mu g/mL, the cisplatin has a good effect of inhibiting cell growth of human bile duct cancer QBC939, and mainly has severe digestive tract reactions such as nausea and vomiting, renal toxicity such as renal tubular injury, and more common neurotoxicity such as tinnitus and hearing loss caused by auditory nerve damage. Because biliary duct cancer has hidden onset and difficult early diagnosis and has late clinical symptoms, epidemiological studies show that only 10-15% of patients are likely to be cured by operation, and many patients find that the patients are in late stage, but the postoperative recurrence rate is still 50-60%, and the five-year survival rate is only 30%. At present, the treatment principle of the bile duct cancer still adopts a comprehensive treatment scheme mainly based on surgical resection, but only patients with deficiency of 1/3 can be surgically resected when the diagnosis is confirmed, the surgical prognosis is poor, and the combined chemotherapy has no ideal effect on locally advanced or metastatic unresectable bile duct cancer. Therefore, the development of new therapeutic drugs is urgently required.

Pancreatic cancer is a common pancreatic tumor, is a digestive tract malignant tumor with high malignancy degree and difficult diagnosis and treatment, and has 5-year survival rate of less than 1 percent. The early diagnosis rate of pancreatic cancer is low, the operative mortality rate is high, and the early diagnosis rate is one of the worst malignant tumors. At present, the main therapeutic drugs comprise fluorouracil, doxorubicin, ifosfamide, semustine, paclitaxel, gemcitabine, cisplatin and the like, but the clinical cure rate is very low, so that research and development of anti-pancreatic cancer drugs with better curative effect are very necessary.

Diosgenin (CAS number 512-06-1) is a steroid compound obtained by hydrolyzing and deglycosylating dioscin contained in rhizome of Dioscoreaceae plants such as Dioscorea zingiberensis, Dioscorea nipponica, Dioscorea panthaica and the like, and is an important raw material for synthesizing steroid hormone drugs and steroid contraceptives. Dioscin has certain anticancer activity (application of protodioscin in preparation of drug-resistant osteosarcoma medicine CN201810091212.3), has activity on lung cancer cells A549(Wei Y, et al. Anti-cancer effects of Dioscin on third genes of human lung cancer cells line identification DNA damage and activating mitotic cancer therapy, 2013, 59), stomach cancer cells MGC-803 (Zhuao X, et al. patent effects of diosgenin targeting cancer in vitro and vivo. phytochrome therapy, 2016, 23), liver cancer cells Hu 7(Hsieh M J, et al. autoinjection therapy, apoptosis induced breast cancer cells 7. basic, and cancer cells ROS, and has activity on lung cancer cells A549 (cancer cells ROS, tissue culture cell ROS, see 3, 2016. 3, 23), liver cancer cells Hu-7 (tumor diagnosis cells, mouse tissue culture medium J, et al. apoptosis inducing hormone, cancer cells, calcium cancer, diosgenin also has certain activity on sarcoma-180, hepatoma ascites type, mouse cervical cancer and ehrlich ascites carcinoma (new medicinal use of diosgenin CN01129317.9), and diosgenin 3-site amide nitrogen-containing derivative has certain antitumor activity (a diosgenin antitumor derivative and its synthesis method CN 201711083013; Geng Qian, et al. Huaxi J. Pharmacology, 2009,24, 475; Jianhongping, et al. Pharmacology, 2011,46, 539; Yingchun Yao, et al. Huaxi J. Pharmacology, 2011,26, 423; Dingyong, et al. Pharmacology, 2012,46, 539; Wuyaki, et al. Huaxi J. Pharmacology, 2012,27,22), and diosgenin 3-site derivative can inhibit ocular abnormal angiogenesis (use of diosgenin-3-site derivative CN201310325890.9), and amino acid diosgenin ester has anti-inflammatory and antitumor effects (diosgenin-3-site derivative CN201310325890.9), The preparation method and the application of CN 201210303318.8; foudao, et al, journal of western medicine, 2010,25,658), diosgenin amino acid derivatives have growth inhibitory effects to different degrees on human colon cancer cells Caco-2, human gastric adenocarcinoma cells, human lung adenocarcinoma cells, and human neuroblastoma cells (diosgenin amino acid derivatives and their use in preparing anti-tumor drugs CN 201410250801.3; the food safety quality detection report 2015,3,1061), diosgenin 3-glycoside such as pentaglycoside and disaccharide, has certain inhibiting effect on breast cancer cells and human liver cancer cells.

Disclosure of Invention

Specifically, the invention has the following technical scheme:

in a first aspect of the invention, the invention provides a compound having the structure shown in formula (I):

wherein R is an unsubstituted or substituted nitrogen-containing heterocyclic group; the nitrogen heterocyclic group is a five-membered or six-membered nitrogen-containing aliphatic heterocyclic ring or a nitrogen-containing aromatic heterocyclic ring; the substituent is selected from alkyl, halogenated alkyl, carboxyl and halogen; the halogen is selected from F, Cl, Br and I.

Also, the present invention provides pharmaceutically acceptable salts of the compounds of formula (I).

Examples of the "pharmaceutically acceptable salt" include, for example, inorganic acid salts such as hydrochloride, hydrobromide, sulfate, phosphate and nitrate; organic acid salts such as acetate, propionate, oxalate, succinate, lactate, malate, tartrate, citrate, maleate, fumarate, methanesulfonate, benzenesulfonate, p-toluenesulfonate and ascorbate; inorganic base salts such as sodium salt, potassium salt, calcium salt, zinc salt, magnesium salt and aluminum salt; and organic base salts such as arginine salt, benzathine salt, choline salt, diethylamine salt, dialcohol amine salt, glycinate salt, lysine salt, meglumine salt, ethanolamine salt, and tromethamine salt.

In an embodiment of the invention, the nitrogen-containing heterocycle is selected from pyrrole, thiazole, imidazole, pyrazole, oxazole, isoxazole, pyridine, pyrimidine, pyridazine, pyrazine and piperidine.

In an embodiment of the invention, the substituents are selected from C_1-3Alkyl, trifluoromethyl and carboxyl; the alkyl group is in particular methyl.

In embodiments of the invention the substitution is one or more substitutions, for example, in some embodiments of the invention the nitrogen-containing heterocycle is isoxazole, which is disubstituted with methyl in the positions C-3 and C-5 respectively of isoxazole, then R is 3, 5-dimethylisoxazol-4-yl, which can be represented by

The nitrogen-containing heterocycle is isoxazole, which is trisubstituted by methyl, the substituted positions are respectively C-3 position, C-4 position and C-5 position of the isoxazole, and R is 3, 5-dimethyl isoxazolyl-4-methyl and can be expressed as

In some embodiments of the invention, the nitrogen-containing heterocycle is unsubstituted, then R is selected from 2-pyrrolyl, 3-pyrrolyl, 2-imidazolyl, 4-imidazolyl, 3-pyrazolyl, 4-pyrazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 3-pyridazinyl, 4-pyridazinyl, and 2-pyrazinyl.

In some embodiments of the invention, R is selected from the group consisting of: 2-pyrrolyl, 3-pyrrolyl, 2-methyl-4-pyrrolyl, 2-imidazolyl, 4-methyl-2-imidazolyl, 2-methyl-4-imidazolyl, 3-pyrazolyl, 4-pyrazolyl, 5-methyl-3-pyrazolyl, 3-methyl-4-pyrazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 2-oxazolylmethyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 4-isoxazolyl, 3, 5-dimethylisoxazolyl-4-methyl, 2-imidazolyl, 4-pyrazolyl, 4-thiazolyl, 5-thiazolyl, 2-thiazolyl, 4-oxazolyl, 3-dimethylisoxazolyl-4-methyl, 2-oxazolyl, 3-dimethylisoxazolyl, 4-methyl, and mixtures thereof, 2-pyridyl group, 3-pyridyl group, 4-pyridyl group, 3-methyl-2-pyridyl group, 4-methyl-2-pyridyl group, 5-methyl-2-pyridyl group, 2-methyl-3-pyridyl group, 4-methyl-3-pyridyl group, 5-methyl-3-pyridyl group, 2-methyl-4-pyridyl group, 3-trifluoromethyl-2-pyridyl group, 4-trifluoromethyl-2-pyridyl group, 5-trifluoromethyl-2-pyridyl group, 2-trifluoromethyl-3-pyridyl group, 4-trifluoromethyl-3-pyridyl group, 5-trifluoromethyl-3-pyridyl group, 3-trifluoromethyl-pyridyl group, 3-pyridyl group, 4-trifluoromethyl-2-pyridyl group, 3-pyridyl group, 4-trifluoromethyl-pyridyl group, 3-, 2-trifluoromethyl-4-pyridyl group, 3-carboxy-2-pyridyl group, 4-carboxy-2-pyridyl group, 5-carboxy-2-pyridyl group, 2-carboxy-3-pyridyl group, 4-carboxy-3-pyridyl group, 5-carboxy-3-pyridyl group, 2-carboxy-4-pyridyl group, 3-pyridylmethyl group, 4-pyridylmethyl group, 2-pyrimidyl group, 4-pyrimidyl group, 5-pyrimidyl group, 4-methyl-2-pyrimidyl group, 5-methyl-2-pyrimidyl group, 2-methyl-4-pyrimidyl group, 3-carboxy-2-pyridyl group, 5-carboxy-3-pyridyl group, 2-methyl-5-pyrimidinyl, 2-pyrimidylmethyl, 4-pyrimidylmethyl, 5-pyrimidylmethyl, 3-pyridazinyl, 4-methyl-3-pyridazinyl, 5-methyl-3-pyridazinyl, 3-methyl-4-pyridazinyl, 3-pyridazinylmethyl, 4-pyridazinylmethyl, 2-pyrazinyl, 3-methyl-2-pyrazinyl and 2-pyrazinylmethyl.

Further, the compounds of formula (I) of the present invention include the structures shown in table 1:

TABLE 1 Structure of the compound of formula (I) and HR-MS structural analysis thereof

In a second aspect of the invention, there is also provided a process for the preparation of a compound of formula (I), said process comprising reacting diosgenin as an initial reactant with RX to produce a compound of formula (I); wherein R is as defined above for the first aspect of the invention and X is halogen.

In an embodiment of the present invention, the method comprises dissolving diosgenin in a solvent, adding RX to react with an alkaline substance, and keeping the solution alkaline during the reaction to obtain the compound of formula (I).

In the embodiment of the invention, the reaction temperature is-20-300 ℃, and the reaction time is 0.1-72 h; preferably, the reaction temperature is-20 to 120 ℃, at which the reaction proceeds more easily.

In an embodiment of the present invention, the solvent is selected from one or more of water, methanol, ethanol (absolute ethanol is preferred), propanol, butanol, pentanol, glycerol, dichloromethane, chloroform, carbon tetrachloride, benzene, toluene, xylene, 1, 4-dioxane, 1, 2-dichloroethane, tetrahydrofuran, N-Dimethylformamide (DMF), and ethyl acetate and dimethyl sulfoxide (DMSO); preferably any one of toluene, absolute ethanol, methanol, n-propanol, isopropanol, n-butanol, ethyl acetate, butyl acetate, methylene chloride, carbon tetrachloride, DMF and tetrahydrofuran, in which the reaction proceeds more easily.

In an embodiment of the present invention, the solvent is used in an amount of 5-100L per mole of diosgenin dissolved in the solvent.

In an embodiment of the invention, the alkaline substance is selected from γ -Al₂O₃-Na、γ-Al₂O₃-K、γ-Al₂O₃-NaOH-Na、γ-Al₂O₃-NaOH-K、γ-Al₂O₃-KOH-Na、γ-Al₂O₃-one or more of KOH-K, sodium methoxide, sodium ethoxide, sodium propoxide, sodium butoxide, sodium amide, sodium metal, sodium hydride, sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium oxide, potassium oxide, calcium oxide, strontium carbonate, potassium carbonate, sodium carbonate, potassium phosphate, ammonia, ethylamine, diethylamine, triethylamine, methylamine, dimethylamine, trimethylamine, pyridine, and piperidine.

In some preferred embodiments, the alkaline material is sodium hydride, potassium hydride, sodium ethoxide, strontium carbonate, potassium carbonate, γ -Al₂O₃-K, potassium carbonate, triethylamine, sodium tert-butoxide, sodium ethoxide, gamma-Al₂O₃KOH-Na or diethylamine, the reaction proceeds more easily in the presence of these basic substances.

In an embodiment of the present invention, the molar ratio of the diosgenin, RX, and the alkaline substance is 1: (0.1-10): (0.01-10), preferably 1 (1.0-1.2): (0.6-1.0).

In some embodiments of the present invention, the weight ratio of the diosgenin, RX and the alkaline substance is (4-20): (1.5-10): (0.01-5).

In an embodiment of the present invention, the preparation method of the compound of formula (I) according to the present invention further comprises detecting the end point of the reaction by chromatography, standing to room temperature after the reaction is finished, and separating and purifying to prepare the compound of formula (I).

In an embodiment of the present invention, the separation and purification method is selected from a combination of one or more of filtration, resin treatment, water washing, distillation, crystallization, extraction, activated carbon treatment, molecular sieve treatment, and chromatography.

The separation and purification method of the invention can be as follows: for example, in some embodiments, TLC detects the total reaction of diosgenin, heats to room temperature, adds water, extracts with dichloromethane, chloroform, acetone or ethyl acetate for 1 to more times, such as 3 times (e.g., 50mL × 3-300mL × 3), follows the reaction and separation and purification process of the product by TLC, recovers the extract, and dries the solid substance, such as at 60 ℃, for 4h to obtain the target product.

For example, in some embodiments, the separation and purification method may be: detecting all reactions of diosgenin by HPLC, distilling off solvent until liquid volume is reduced to 1/4, cooling and crystallizing overnight, such as cooling to 5 deg.C and crystallizing overnight, filtering, tracking reaction and product separation and purification process by HPLC, and drying solid, such as drying at 60 deg.C for 4 hr, to obtain target product.

For example, in some further embodiments, the separation and purification method may be: detecting the total reaction of diosgenin by HPLC, reducing pressure to remove solvent, performing chromatography, such as chromatography with chromatography column, eluting with ethyl acetate-ethanol (v/v ═ 10:1) or dichloromethane-methanol (v/v ═ 5:1), tracking the separation and purification process of reaction and product by HPLC, collecting and combining product fractions, and evaporating solvent by rotary evaporator (such as at 60 deg.C) to obtain the target product.

In a third aspect of the invention, the present invention also provides a composition comprising a compound of formula (I) as described in the above first aspect of the invention or a pharmaceutically acceptable salt thereof.

In a fourth aspect of the invention, the invention also provides a pharmaceutical formulation comprising a compound of formula (I) as described in the above first aspect of the invention or a pharmaceutically acceptable salt thereof or a composition comprising said compound of formula (I) or a salt thereof, together with at least one pharmaceutical carrier or adjuvant.

The pharmaceutical compositions of the present invention are generally safe, non-toxic and biologically desirable, the pharmaceutically acceptable carriers or excipients of the present invention are non-toxic and safe, and their combinations with the compounds of the present invention are also non-toxic and safe. The pharmaceutically acceptable carriers and excipients of the present invention are generally well known to those skilled in the art or can be determined by those skilled in the art based on the actual circumstances. Examples of suitable carriers and excipients include glucose, water, glycerol, ethanol, propylene glycol, corn starch, gelatin, lactose, sucrose, alginic acid, microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride, croscarmellose sodium and sodium starch glycolate and the like polysorbate 80, polyethylene glycol 300, polyethylene glycol 400, cyclodextrins or derivatives thereof such as ((2-hydroxypropyl) -cyclodextrin) and (2-hydroxyethyl) -cyclodextrin, which are also known as HPCD, pegylated castor oil, poloxamers such as poloxamer 407 or 188; hydrophilic carriers, hydrophobic carriers, or combinations thereof, and the like. Hydrophobic carriers include, for example, fat emulsions, lipids, pegylated phospholipids, biocompatible polymers, lipid spheres, liposomes, vesicles, polymer matrices, particles, and the like. Furthermore, one skilled in the art will appreciate that diluents are included within the term carrier and adjuvant.

The amount of carrier or adjuvant in the pharmaceutical composition may be from 1% to 98% by weight, usually about 80% by weight. For convenience, the local anesthetic, preservative, buffer, etc. may be dissolved directly in the vehicle.

Pharmaceutical compositions of the compounds of the present invention may be administered in any manner selected from: oral, aerosol inhalation, rectal, nasal, vaginal, topical, parenteral such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal or intracranial injection or infusion, or by means of an explanted reservoir, with oral, intramuscular, intraperitoneal or intravenous administration being preferred. The pharmaceutical dosage form can be a liquid dosage form, a solid dosage form. The liquid dosage form can be true solution, colloid, microparticle, emulsion, or mixed suspension. Other dosage forms such as tablet, capsule, dripping pill, aerosol, pill, powder, solution, suspension, emulsion, granule, suppository, lyophilized powder for injection, clathrate, landfill, patch, liniment, etc.

In a fifth aspect, the present invention also provides the use of a compound of formula (I) as described in the above first aspect of the present invention or a pharmaceutically acceptable salt thereof, or a composition comprising a compound of formula (I) or a salt thereof, or a pharmaceutical formulation comprising a compound of formula (I) or a salt thereof, in the manufacture of a medicament for the treatment of cancer; in an embodiment of the invention, the cancer is preferably pancreatic cancer and/or cholangiocarcinoma.

In some embodiments of the invention, the inhibition rate of 0.2ug/mL of the compound of formula (I) of the invention on the proliferation of bile duct cancer cells QBC-939 is higher than 70%, and can reach 95%; the inhibition rate of 0.2ug/mL of the compound shown in the formula (I) on the proliferation of human pancreatic cancer cells PANC-1 is higher than 70% and can reach 95%. The activity of the compound of the invention is far higher than that of 5-fluorouracil used clinically.

Detailed Description

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. The reagents or starting materials used in the present invention can be purchased from conventional sources, and unless otherwise specified, the reagents or starting materials used in the present invention can be used in a conventional manner in the art or in accordance with the product specifications. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.

Example 1

Weighing 4.2g of diosgenin and 0.5g of sodium hydride, dissolving in 500mL of 100mL of absolute ethanol, adding 1.5g of 2-chloropyrrole, stirring and freezing to-20 ℃, keeping the temperature and stirring for reaction for 72h, detecting the total reaction of the diosgenin by TLC (a developing agent: petroleum ether-ethyl acetate (1:1) and a color developing agent: 5% phosphomolybdic acid ethanol solution), distilling out the ethanol, cooling to room temperature, adding 50mL of water, extracting for 3 times (60mL multiplied by 3) by dichloromethane, recovering the dichloromethane from an extract, tracking the reaction by TLC and the separation and purification process of a product, and drying the solid at 60 ℃ for 4h to obtain 3.2g of a light yellow powdery product. Melting point of the target product: 192.5-194.5 ℃ of the total weight of the mixture,¹³C NMR(CDCl₃100MHz) delta 143.56,140.83,121.85,119.64,118.34,108.61,107.92,82.95,82.33,66.94,62.82,56.64,50.81,42.15,40.71,40.13,39.24,37.73,37.52,32.86,32.26,32.05,31.31,30.35,29.14,28.86,21.13,19.37,17.56,15.50, 14.82. TOF-HRMS: m/e (479.6925), formula: c₃₁H₄₅O₃N, i.e. a compound in Table 1Object 1.

Compound 2(R is 3-pyrrolyl) can be prepared in a similar manner to example 1, using the appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 2

Weighing diosgenin 4.2g and potassium hydride 1.0g, dissolving in 100mL of methanol 500mL, adding 2-methyl-3-chloro-pyrrole 1.5g, stirring and heating at 25 ℃, keeping the temperature and stirring for reaction for 36h, detecting the total reaction of diosgenin by TLC (developing agent: petroleum ether-ethyl acetate (1:1), color developing agent: 5% phosphomolybdic acid ethanol solution), evaporating out methanol, cooling to room temperature, adding water 50mL, extracting with dichloromethane for 3 times (60mL multiplied by 3), recovering dichloromethane from extract liquid, tracking the reaction by TLC and the separation and purification process of the product, and drying the solid at 60 ℃ for 4h to obtain light yellow powder product 3.4 g. Melting point of the target product: 191.7-193.9 ℃ of the total weight of the composition,¹³C NMR(CDCl₃100MHz) delta 140.80,129.86,121.82,119.61,118.05,108.21,107.83,82.92,82.50,66.91,62.79,56.61,50.78,42.12,40.68,40.10,39.21,37.70,37.49,32.83,32.20,32.02,31.28,30.32,29.11,28.83,21.10,19.34,17.53,15.47,14.79, 11.95. TOF-HRMS: m/e (493.7189), formula: c₃₂H₄₇O₃N, compound 3 in table 1.

Compound 4(R ═ 2-methyl-4-pyrrolyl) can be prepared according to a similar procedure to example 2, using the appropriate halogenated nitrogen-containing heterocyclyl group (halogen can be Cl or Br).

Example 3

Weighing diosgenin 4.2g and sodium ethoxide 0.8g, dissolving in 100mL of n-propanol in a 500mL three-neck flask, adding 2-chloroimidazole 1.5g, stirring and heating at 75 ℃, keeping the temperature and stirring for reaction for 48h, detecting the total reaction of the diosgenin by TLC (a developing agent: petroleum ether-ethyl acetate (1:1), a color developing agent: 5% phosphomolybdic acid ethanol solution), distilling out the n-propanol, cooling to room temperature, separating by using a silica gel chromatographic column, eluting by using ethyl acetate-ethanol (such as 10:1), collecting a product fraction, carrying out rotary evaporation on the solvent to obtain a solid, tracking the separation and purification processes of the reaction and the product by TLC, and drying at 60 ℃ for 4h to obtain a light yellow powdery product 3.5 g. Melting point of the target product: 195.1-196.8 deg.C，¹³C NMR(CDCl₃100MHz) delta 150.11,140.81,127.84,127.81,121.83,119.62,82.93,81.28,66.92,62.80,56.62,50.79,42.13,40.69,40.11,39.22,37.71,37.50,32.84,32.21,32.03,31.29,30.33,29.12,28.84,21.11,19.35,17.54,15.48, 14.80. TOF-HRMS: m/e (480.6806), formula: c₃₀H₄₄O₃N₂I.e. compound 5 in table 1.

Compound 6(R ═ 4-imidazolyl) can be prepared in a similar manner to example 3, using the appropriate halogenated nitrogen-containing heterocyclic group (halogen can be Cl or Br).

Example 4

Weighing diosgenin 4.2g and strontium carbonate 0.05g, dissolving in 100mL n-propanol in 500mL three-neck flask, adding 2-methyl-4-bromo-imidazole 2.0g, heating and stirring at 95 deg.C, maintaining the temperature and stirring for reaction for 18h, detecting total reaction of diosgenin by TLC, evaporating solvent under reduced pressure, cooling to room temperature, adding 50mL water, extracting with acetone for 3 times (50mL × 3), extracting with anhydrous Na₂SO₄Drying for 8h, filtering, recovering acetone from the filtrate to obtain solid, tracking reaction by TLC, separating and purifying the product, and drying the solid at 60 deg.C for 4h to obtain light yellow powder 3.7 g. Melting point of the target product: the temperature of the mixture is 194.5 to 196.6 ℃,¹³C NMR(101MHz,CDCl₃) δ 145.72,140.82,122.25,122.04,121.84,119.63,82.94,81.62,66.93,62.81,56.63,50.80,42.14,40.70,40.12,39.23,37.72,37.51,32.85,32.22,32.04,31.30,30.34,29.13,28.85,21.12,19.36,17.95,17.55,15.49, 14.81. TOF-HRMS: m/e (494.7071), formula: c₃₁H₄₆O₃N₂I.e. compound 8 in table 1.

Compound 7(R ═ 4-methyl-2-imidazolyl) was prepared in a similar manner to example 4, using the appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 5

Weighing diosgenin 4.2g and potassium carbonate 1.5g, dissolving in carbon tetrachloride 100mL in 500mL three-neck flask, adding 4-bromo-pyrazole 2.0g, stirring and heating to boil (97 deg.C), maintaining the temperature and stirring for reaction for 16h, detecting by TLC that diosgenin completely reacts, distilling under reduced pressure to remove dissolved diosgeninCooling to room temperature, adding 100mL of water, adsorbing by 101 macroporous resin, eluting with 95% of water, collecting product fractions, recovering ethanol to obtain a solid, tracking reaction by TLC and separating and purifying the product, and drying the solid at 60 ℃ for 4h to obtain 2.9g of a light yellow powdered product. Melting point of the target product: the temperature of the mixture is 195.4-197.3 ℃,¹³C NMR(100MHz,CDCl₃) δ 141.23,140.93,121.95,119.74,114.45,114.44,83.05,81.25,67.04,62.92,56.74,50.91,42.25,40.81,40.23,39.34,37.83,37.62,32.96,32.33,32.15,31.41,30.45,29.24,28.96,21.23,19.47,17.66,15.60, 14.92. TOF-HRMS: m/e (480.6802), formula: c₃₀H₄₄O₃N₂Compound 10 in table 1.

Compound 9(R ═ 3-pyrazolyl) can be prepared according to a similar manner to example 5, using an appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 6

Weighing diosgenin 4.2g and gamma-Al₂O₃Dissolving 0.5g of-K in 100mL of isopropanol in a 500mL three-neck flask, adding 2.2g of 5-methyl-3-bromo-pyrazole, heating and stirring at 98 ℃, keeping the temperature and stirring for reaction for 24 hours, detecting all reactions of diosgenin by TLC (developing agent: petroleum ether-ethyl acetate (1:1) and color developing agent: 5% phosphomolybdic acid ethanol solution), distilling the solvent under reduced pressure, cooling to room temperature, adding 50mL of water, extracting for 3 times (50mL × 3) with acetone, and extracting the extract with anhydrous Na₂SO₄Drying for 8h, filtering, recovering acetone from the filtrate to obtain solid, tracking reaction by TLC, separating and purifying the product, and drying the solid at 60 deg.C for 4h to obtain light yellow powder 3.2 g. Melting point of the target product: 192.5-193.9 ℃ of the weight percentage,¹³C NMR(101MHz,CDCl₃) δ 169.15,141.95,140.94,121.96,119.75,97.24,83.06,81.15,67.05,62.93,56.75,50.92,42.26,40.82,40.24,39.35,37.84,37.63,32.97,32.34,32.16,31.42,30.46,29.25,28.97,21.24,19.48,17.67,15.61,14.93, 13.46. TOF-HRMS: m/e (494.7071), formula: c₃₁H₄₆O₃N₂I.e. compound 11 in table 1.

Compound 12(R ═ 3-methyl-4-pyrazolyl) can be prepared according to a similar manner to that of example 6, using an appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 7

Weighing diosgenin 4.2g and potassium carbonate 1.5g, dissolving in 100mL n-butanol in 500mL three-neck flask, adding 2-bromo-thiazole 1.8g, heating and stirring at 100 deg.C, maintaining the temperature and stirring for reaction for 12h, detecting total reaction of diosgenin by TLC (developing agent: petroleum ether-ethyl acetate (1:1), developer: 5% phosphomolybdic acid ethanol solution), distilling off solvent under reduced pressure, cooling to room temperature, adding water 50mL, extracting with acetone for 3 times (50mL × 3), extracting with anhydrous Na₂SO₄Drying for 8h, filtering, recovering acetone from the filtrate to obtain solid, tracking reaction by TLC, separating and purifying the product, and drying the solid at 60 deg.C for 4h to obtain light yellow powder 3.9 g. Melting point of the target product: 195.7-197.5 ℃ of the total weight of the feed,¹³C NMR(101MHz,CDCl₃) δ 154.12,142.65,140.91,121.93,119.72,118.64,83.03,81.15,67.02,62.90,56.72,50.89,42.23,40.79,40.21,39.32,37.81,37.60,32.94,32.31,32.13,31.39,30.43,29.22,28.94,21.21,19.45,17.64,15.58, 14.90. TOF-HRMS: m/e (497.7311), formula: c₃₀H₄₃O₃NS, compound 13 in table 1.

Compound 14(R ═ 4-thiazolyl) and compound 15(R ═ 5-thiazolyl) can be prepared according to a similar procedure to example 7, using the appropriate halogenated nitrogen-containing heterocyclic group (halogen can be Cl or Br), respectively.

Example 8

Weighing 4.2g of diosgenin and 1.5g of potassium carbonate, dissolving in 100mL of toluene in a 500mL three-neck flask, adding 1.8g of 4-bromo-oxazole, heating and stirring for 90 ℃, keeping the temperature and stirring for reaction for 12h, detecting the total reaction of the diosgenin by TLC (a developing agent: petroleum ether-ethyl acetate (1:1), a color developing agent: 5% phosphomolybdic acid ethanol solution), distilling the solvent out under reduced pressure, cooling to room temperature, adding 100mL of water, adsorbing by 101 macroporous resin, eluting by 95% of ethanol, collecting product fractions, distilling the solvent to obtain a solid, tracking the separation and purification process of the reaction and the product by TLC, drying the solid at 60 ℃ for 4h to obtain 4.0g of a light yellow powdery product. Melting point of the target product: the temperature of the mixed solution is between 191.4 and 193.3 ℃,¹³C NMR(101MHz,CDCl₃) δ 151.43,140.92,138.14,125.62,121.94,119.73,83.04,81.19,67.03,62.91,56.73,50.90,42.24,40.80,40.22,39.33,37.82,37.61,32.95,32.32,32.14,31.40,30.44,29.23,28.95,21.22,19.46,17.65,15.59, 14.91. TOF-HRMS: m/e (481.6651), formula: c₃₀H₄₃O₄N, compound 17 in table 1.

Compound 16(R ═ 2-oxazolyl) and compound 18(R ═ 5-oxazolyl) can be prepared according to a similar procedure to that of example 8, using the appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 9

Weighing diosgenin 4.2g and potassium carbonate 1.5g, dissolving in tetrahydrofuran 100mL in a 500mL three-neck flask, adding 2-oxazolylmethyl bromide 2.0g, heating and stirring at 50 deg.C, maintaining the temperature and stirring for reaction for 12h, detecting diosgenin by TLC (developing agent: petroleum ether-ethyl acetate (1:1), developer: 5% phosphomolybdic acid ethanol solution), reacting, evaporating solvent under reduced pressure, cooling to room temperature, adding water 50mL, extracting with acetone for 3 times (50mL × 3), extracting with anhydrous Na₂SO₄Drying for 8h, filtering, recovering acetone from the filtrate to obtain solid, tracking reaction by TLC, separating and purifying the product, and drying the solid at 60 deg.C for 4h to obtain light yellow powder 3.7 g. Melting point of the target product: the temperature of the mixture is between 190.4 and 192.1 ℃,¹³C NMR(101MHz,CDCl₃) δ 150.62,140.89,139.70,127.06,121.91,119.70,87.15,83.06,67.81,67.00,62.88,56.70,50.87,42.21,40.77,40.19,39.30,37.79,37.58,32.92,32.29,32.11,31.37,30.41,29.30,28.92,21.19,19.43,17.62,15.56, 14.88. TOF-HRMS: m/e (495.6918), formula: c₃₁H₄₅O₄N, compound 19 in table 1.

Example 10

Weighing diosgenin 4.2g, dissolving in ethyl acetate 80mL in a three-neck flask 500mL, adding triethylamine 10mL and 4-chloro-isoxazole 1.8g, stirring, freezing to-10 deg.C, reacting for 72h under heat preservation, detecting diosgenin by TLC (developing agent: petroleum ether-ethyl acetate (1:1), developer: 5% phosphomolybdic acid ethanol solution), heating to room temperature, keeping the temperature for 2h, and steaming under reduced pressure until the liquid volume is 20mL, cooling to 5 ℃, crystallizing overnight, filtering, TLC tracing reaction and separation and purification process of the product, drying the solid at 60 ℃ for 4h to obtain 2.9g of product of light yellow powder. Melting point of the target product: the temperature of the mixture is between 190.2 and 192.0 ℃,¹³C NMR(101MHz,CDCl₃) δ 158.83,150.07,140.87,121.89,119.68,100.56,83.04,82.10,66.98,62.86,56.68,50.85,42.19,40.75,40.17,39.28,37.77,37.56,32.90,32.27,32.09,31.35,30.39,29.28,28.90,21.17,19.41,17.60,15.54, 14.86. TOF-HRMS: m/e (481.6647), formula: c₃₀H₄₃O₄N, compound 21 in table 1.

Compound 20(R ═ 3-isoxazolyl), compound 22(R ═ 5-isoxazolyl), and compound 23(R ═ 4-isoxazolylmethyl) were each prepared according to a similar procedure to that of example 10, using an appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 11

Weighing 4.2g of diosgenin, dissolving in 80mL of DMF (dimethyl formamide) in a 500mL three-neck flask, adding 10mL of triethylamine and 2.0g of 3, 5-dimethyl-4-chloromethyl isoxazole, stirring, freezing to 0 ℃, preserving heat for reaction for 36h, detecting the total reaction of the diosgenin by TLC (developing agent: petroleum ether-ethyl acetate (1:1) and color developing agent: 5% phosphomolybdic acid ethanol solution), heating to room temperature, preserving heat for 2h, evaporating under reduced pressure until the liquid volume is 20mL, cooling to 5 ℃, crystallizing overnight, filtering, tracking the separation and purification process of the reaction and the product by TLC, and drying the solid at 60 ℃ for 4h to obtain 3.3g of light yellow powder product. Melting point of the target product: at the temperature of 189.5-191.2 ℃,¹³C NMR(101MHz,CDCl₃) δ 158.92,150.05,140.85,121.87,119.66,100.50,83.02,81.85,66.96,62.84,56.66,50.83,42.17,40.73,40.15,39.26,37.75,37.54,32.88,32.25,32.07,31.33,30.37,29.26,28.88,21.15,19.39,17.58,15.52,14.84,10.61, 9.27. TOF-HRMS: m/e (523.7451), formula: c₃₃H₄₉O₄N, compound 24 in table 1.

Example 12

Weighing diosgenin 4.2g, dissolving in 80mL butyl acetate in 500mL three-neck flask, adding sodium ethoxide 1.0g and 2-chloro-pyridine 2.0g, stirring, reacting at room temperature for 24 hr, and TLC (developing solvent)Detecting the total reaction of diosgenin by using petroleum ether-ethyl acetate (1:1) and a color developing agent which is 5% phosphomolybdic acid ethanol solution), steaming under reduced pressure until the volume of liquid is 20mL, cooling to 5 ℃, crystallizing overnight, filtering, tracing the reaction by TLC and separating and purifying the product, and drying the solid at 60 ℃ for 4 hours to obtain 3.4g of light yellow powder product. Melting point of the target product: 192.5-194.3 ℃ of the total weight of the mixture,¹³C NMR(101MHz,CDCl₃) δ 163.42,146.81,140.84,138.10,121.86,119.65,116.75,109.21,83.01,81.54,66.95,62.83,56.65,50.82,42.16,40.72,40.14,39.25,37.74,37.53,32.87,32.24,32.06,31.32,30.36,29.25,28.87,21.14,19.38,17.57,15.51, 14.83. TOF-HRMS: m/e (491.7038), formula: c₃₂H₄₅O₃N, compound 25 in table 1.

Compound 26(R ═ 3-pyridyl) and compound 27(R ═ 4-pyridyl) can be prepared according to a similar manner to example 12, using an appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 13

Weighing 4.2g of diosgenin, dissolving in 200mL of dichloromethane in a 500mL three-neck flask, adding 1.8g of sodium tert-butoxide and 2.5g of 2-methyl-4-iodo-pyridine, stirring, reacting at room temperature for 24h, detecting all reactions of the diosgenin by TLC (developing agent: petroleum ether-ethyl acetate (1:1) and developer: 5% phosphomolybdic acid ethanol solution), evaporating under reduced pressure until the liquid volume is 20mL, cooling to 5 ℃, crystallizing overnight, filtering, tracking the reaction and the separation and purification process of the product by TLC, and drying the solid at 60 ℃ for 4h to obtain 4.0g of a light yellow powder product. Melting point of the target product: the temperature of the mixed solution is between 191.5 and 193.3 ℃,¹³C NMR(101MHz,CDCl₃) δ 158.91,158.30,149.52,140.80,121.82,119.61,112.95,105.52,82.97,81.62,66.91,62.79,56.61,50.78,42.12,40.68,40.10,39.21,37.70,37.49,32.83,32.20,32.02,31.28,30.32,29.21,28.83,24.22,21.10,19.34,17.53,15.46, 14.79. TOF-HRMS: m/e (505.7306), formula: c₃₃H₄₇O₃N, compound 34 in table 1.

Compound 28(R ═ 3-methyl-2-pyridyl), compound 29(R ═ 4-methyl-2-pyridyl), compound 30(R ═ 5-methyl-2-pyridyl), compound 31(R ═ 2-methyl-3-pyridyl), compound 32(R ═ 4-methyl-3-pyridyl), compound 33 (5-methyl-3-pyridyl), and compound 35(R ═ 3-methyl-4-pyridyl) can be prepared according to a similar method to example 13, using an appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 14

Weighing diosgenin 4.2g and sodium ethoxide 0.8g, dissolving in 100mL of n-propanol in a 500mL three-neck flask, adding 3-trifluoromethyl-2-bromo-pyridine 2.5g, stirring and heating at 60 ℃, keeping the temperature and stirring for reaction for 36h, detecting the total reaction of the diosgenin by TLC (a developing agent: petroleum ether-ethyl acetate (1:1) and a color developing agent: 5% phosphomolybdic acid ethanol solution), evaporating the n-propanol, cooling to room temperature, adding 80mL of water, extracting for 3 times (100mL multiplied by 3) by using trichloromethane, tracking the reaction by TLC and the separation and purification process of a product, recovering the trichloromethane from an extract, and drying a solid at 60 ℃ for 4h to obtain a light yellow powdery product 3.0 g. Melting point of the target product: the temperature of the mixture is between 201.1 and 202.7 ℃,¹³C NMR(CDCl₃100MHz) delta 159.90,144.52,140.90,137.23,125.15,121.92,119.71,118.23,108.61,83.07,81.54,67.01,62.89,56.71,50.88,42.22,40.78,40.20,39.31,37.80,37.59,32.93,32.30,32.12,31.38,30.42,29.31,28.93,21.20,19.44,17.63,15.56, 14.89. TOF-HRMS: m/e (559.7014), formula: c₃₃H₄₄O₃NF₃Compound 36 in table 1.

Compound 37(R ═ 4-trifluoromethyl-2-pyridyl) and compound 38(R ═ 5-trifluoromethyl-2-pyridyl) can be prepared according to a similar procedure to that of example 14, using an appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 15

Weighing diosgenin 4.2g and potassium carbonate 1.5g, dissolving in 100mL isopropanol in 500mL three-neck flask, adding 2-trifluoromethyl-4-bromo-pyridine 2.6g, heating and stirring at 110 deg.C, maintaining the temperature and stirring for reaction for 12h, detecting total reaction of diosgenin by TLC (developing agent: petroleum ether-ethyl acetate (1:1), developer: 5% phosphomolybdic acid ethanol solution), distilling off solvent under reduced pressure, cooling to room temperature, adding water 50mL, extracting with acetone for 3 times (50mL × 3), tracking reaction by TLC, and separating and purifying productThe extract is extracted with anhydrous Na₂SO₄Drying for 8h, filtering, recovering acetone from the filtrate to obtain solid, and drying at 60 deg.C for 4h to obtain light yellow powder 3.3 g. Melting point of the target product: 102.9-1204.5 ℃ of the weight percentage,¹³C NMR(100MHz,CDCl₃) δ 158.33,149.50,144.62,140.88,120.10,121.90,119.69,112.93,105.65,83.05,81.57,66.99,62.87,56.69,50.86,42.20,40.76,40.18,39.29,37.78,37.57,32.91,32.28,32.10,31.36,30.40,29.29,28.91,21.18,19.42,17.61,15.54, 14.87. TOF-HRMS: m/e (559.7030), formula: c₃₃H₄₄O₃NF₃I.e. compound 42 in table 1.

Compound 39(R ═ 2-trifluoromethyl-3-pyridyl), compound 40(R ═ 4-trifluoromethyl-3-pyridyl), compound 41(R ═ 5-trifluoromethyl-3-pyridyl), and compound 43(R ═ 3-trifluoromethyl-4-pyridyl) can be prepared according to a similar procedure to that of example 15, using an appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 16

Weighing diosgenin 4.2g and gamma-Al₂O₃KOH-Na 0.5g was dissolved in 100mL of isopropanol in a 500mL three-necked flask, and 3-carboxy-2-chloro-pyridine 2.0g was added thereto, and the mixture was stirred under heating at 120 ℃ and kept warm for 24 hours, followed by HPLC (conditions for chromatography: Agilent Zorbax SB-C18(4.6 mm. times.150 mm,5um) as a column, acetonitrile-water (90: 10) as a mobile phase, and a flow rate of 1.0mL.min^-1Detecting total reaction of diosgenin at detection wavelength of 203nm and column temperature of 35 deg.C, distilling off solvent under reduced pressure, cooling to room temperature, adding 50mL of water, extracting with acetone for 3 times (50mL × 3), tracking reaction and product separation and purification process by HPLC, and extracting with anhydrous Na₂SO₄Drying for 8h, filtering, recovering acetone from the filtrate to obtain solid, and drying the solid at 60 deg.C for 4h to obtain light yellow powder 3.5 g. Melting point of the target product: 177.5 to 179.2 ℃ of the mixture,¹³C NMR(101MHz,CDCl₃)δ169.35,161.20,149.81,140.89,139.02,121.91,119.70,119.11,110.83,83.06,81.21,66.70,62.88,56.70,50.87,42.21,40.77,40.19,39.30,37.79,37.58,32.92,32.29,32.11,31.37,30.41,29.30,28.92,21.19,19.43,17.62,15.55,14.88。TOF-HRMS：M/e(535.7154) The molecular formula is as follows: c₃₃H₄₅O₅N, compound 44 in table 1.

Compound 45(R ═ 4-carboxy-2-pyridyl), compound 46(R ═ 5-carboxy-2-pyridyl), compound 47(R ═ 2-carboxy-3-pyridyl), compound 48(R ═ 4-carboxy-3-pyridyl), compound 49(R ═ 5-carboxy-3-pyridyl), compound 50(R ═ 2-carboxy-3-pyridyl), compound 51(R ═ 3-carboxy-4-pyridyl) can be prepared according to a similar method to that of example 16, using an appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 17

Weighing diosgenin 4.2g, dissolving in 80mL ethyl acetate in 500mL three-neck flask, adding triethylamine 10mL and 3-chloromethylpyridine 1.5g, stirring, freezing to-10 deg.C, reacting for 72h under constant temperature, and performing HPLC (chromatographic conditions: Agilent Zorbax SB-C18(4.6mm × 150mm,5um) on chromatographic column, acetonitrile-water (90: 10) as mobile phase, and flow rate of 1.0mL.min^-1Detecting the total reaction of diosgenin at the detection wavelength of 203nm and the column temperature of 35 ℃), heating to room temperature, keeping the temperature for 2h, steaming under reduced pressure until the liquid volume is 20mL, cooling to 5 ℃, crystallizing overnight, filtering, tracing the reaction and the separation and purification process of the product by HPLC, and drying the solid at 60 ℃ for 4h to obtain 3.0g of a light yellow powdery product. Melting point of the target product: the temperature of the mixture is between 191.0 and 192.6 ℃,¹³C NMR(100MHz,CDCl₃) δ 150.13,147.31,140.88,134.33,132.63,123.35,121.90,119.69,87.75,83.05,72.85,66.99,62.87,56.69,50.86,42.20,40.76,40.18,39.29,37.78,37.57,32.91,32.28,32.10,31.36,30.40,29.29,28.91,21.18,19.42,17.61,15.54, 14.87. TOF-HRMS: m/e (505.7301), formula: c₃₃H₄₇O₃N, compound 52 in table 1.

Compound 53(R ═ 4-pyridylmethyl) can be prepared in a similar manner to example 17, using the appropriate halogenated nitrogen-containing heterocyclic group (halogen can be Cl or Br).

Example 18

Weighing diosgenin 4.2g, dissolving in dichloromethane 100mL, adding diethylamine 10mL and 4-chloropyrimidine 1.5g, stirring, reacting at 20 deg.C for 18h, and performing HPLC (chromatography)Conditions are as follows: the chromatographic column was Agilent Zorbax SB-C18(4.6 mm. times.150 mm,5um), the mobile phase was acetonitrile-water (90: 10), and the flow rate was 1.0mL.min^-1Detecting the total reaction of diosgenin at the detection wavelength of 203nm and the column temperature of 35 ℃), evaporating dichloromethane until the liquid volume is reduced to 1/4, cooling to 5 ℃, crystallizing overnight, filtering, tracing the separation and purification process of the reaction and the product by HPLC, and drying the solid at 60 ℃ for 4 hours to obtain 3.1g of yellow powdery product. Melting point of the target product: 195.9-197.5 ℃ of the total weight of the feed,¹³C NMR(CDCl₃100MHz) delta 168.43,157.55,157.34,140.93,121.95,119.74,108.72,83.05,81.57,67.04,62.92,56.74,50.91,42.25,40.81,40.23,39.34,37.83,37.62,32.96,32.33,32.15,31.41,30.45,29.24,28.96,21.23,19.47,17.66,15.60, 14.92. TOF-HRMS: m/e (492.6911), formula: c₃₁H₄₄O₃N₂Compound 55 in table 1.

Compound 54(R ═ 2-pyrimidinyl) and compound 56(R ═ 5-pyrimidinyl) can be prepared according to a similar procedure to that of example 18, using the appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 19

Weighing diosgenin 4.2g, dissolving in dichloromethane 100mL, adding diethylamine 10mL and 2-methyl-4-bromo-pyrimidine 2.5g, stirring, reacting at 40 deg.C for 8h, and performing HPLC (chromatography conditions: Agilent Zorbax SB-C18(4.6mm × 150mm,5um) with mobile phase of acetonitrile-water (90: 10) and flow rate of 1.0mL. min^-1Detecting the total reaction of diosgenin at the detection wavelength of 203nm and the column temperature of 35 ℃), evaporating dichloromethane until the liquid volume is reduced to 1/4, cooling to 5 ℃, crystallizing overnight, filtering, tracing the separation and purification process of the reaction and the product by HPLC, and drying the solid at 60 ℃ for 4 hours to obtain 3.0g of yellow powdery product. Melting point of the target product: 192.6-194.4 ℃ of the total weight of the mixture,¹³C NMR(CDCl₃100MHz) delta 168.02,164.95,159.51,140.90,121.92,119.71,105.75,83.02,81.54,67.01,62.89,56.71,50.88,42.22,40.78,40.20,39.31,37.80,37.59,32.93,32.30,32.12,31.38,30.42,29.21,28.93,24.72,21.20,19.44,17.63,15.57, 14.89. TOF-HRMS: m/e (506.7180), formula: c₃₂H₄₆O₃N₂Compound 59 in table 1.

Compound 57(R ═ 4-methyl-2-pyrimidinyl), 58(R ═ 5-methyl-2-pyrimidinyl), and 60(R ═ 2-methyl-5-pyrimidinyl) were prepared according to a similar procedure to that of example 19, using the appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 20

Weighing 16.8g of diosgenin and 0.8g of sodium hydride, dissolving in 400mL of methanol in a 1000mL three-neck flask, adding 5.8g of 2-chloromethylpyrimidine, stirring and heating at 65 ℃, keeping the temperature and stirring for reaction for 6 hours, detecting all reactions of the diosgenin by TLC, distilling out the methanol, cooling to room temperature, adding 150mL of water, extracting for 3 times (200mL multiplied by 3) by using dichloromethane, tracking the separation and purification processes of the reactions and the products by TLC, recovering the dichloromethane from the extract, and drying the solid at 60 ℃ for 4 hours to obtain 12.2g of a light yellow powdery product. Melting point of the target product: the temperature of between 187.6 and 189.3 ℃,¹³C NMR(CDCl₃100MHz) delta 165.42,157.15,157.14,140.89,121.91,119.70,117.83,87.14,83.01,69.72,67.00,62.88,56.70,50.87,42.21,40.77,40.19,39.30,37.79,37.58,32.92,32.29,32.11,31.37,30.41,29.20,28.92,21.19,19.43,17.62,15.56, 14.88. TOF-HRMS: m/e (506.7179), formula: c₃₂H₄₆O₃N₂Compound 61 in table 1.

Compound 62(R ═ 4-pyrimidinylmethyl) can be prepared according to a similar procedure to example 20, using the appropriate halogenated nitrogen-containing heterocyclyl group (halogen can be Cl or Br).

Example 21

Weighing 16.8g of diosgenin and 2.1g of sodium ethoxide, dissolving in 400mL of absolute ethanol, adding 8.0g of 5-chloromethylpyrimidine into a 1000mL three-neck flask, stirring and heating at 78.5 ℃, keeping the temperature, stirring and reacting for 4 hours, detecting all reactions of the diosgenin by TLC, distilling off the absolute ethanol, cooling to room temperature, adding 120mL of water, extracting for 3 times (200mL multiplied by 3) by using trichloromethane, tracking the separation and purification processes of the reactions and the products by TLC, recovering the trichloromethane from the extract, and drying the solid at 60 ℃ for 4 hours to obtain 13.5g of a light yellow powdery product. Melting point of the target product: 183.5-185.3 ℃ of the total weight of the mixture,¹³C NMR(CDCl₃,100MHz)δ165.62,147.55,140.78,128.61,121.80,119.59,117.33,82.90,81.51,66.89,62.77,56.59,50.76,42.10,40.66,40.08,39.19,37.68,37.47,32.81,32.18,32.00,31.26,30.30,29.09,28.81,21.08,19.32,17.51,15.45,14.77. TOF-HRMS: m/e (506.7180), formula: c₃₂H₄₆O₃N₂Compound 63 in table 1.

Example 22

Weighing 4.2g of diosgenin and 1.40g of potassium carbonate, dissolving in 100mL of propanol in a 500mL three-necked flask, adding 2.1g of 4-methyl-3-bromo-pyridazine, heating and stirring at 98 ℃, keeping the temperature and stirring for reaction for 4h, detecting by TLC (thin layer chromatography) all reactions of the diosgenin, distilling the solvent under reduced pressure, cooling to room temperature, adding 50mL of water, extracting for 3 times (100mL multiplied by 3) with acetone, carrying out TLC (thin layer chromatography) tracking reaction and separation and purification processes of a product, eluting the extract by a silica gel chromatographic column and dichloromethane-absolute ethyl alcohol (5: 1), collecting a product fraction, distilling the solvent, and drying the solid at 60 ℃ for 4h to obtain 3.5g of a light yellow powdery product. Melting point of the target product: 184.5-186.2 ℃ of water,¹³C NMR(101MHz,CDCl₃) δ 159.81,140.79,136.65,131.11,128.03,121.81,119.60,82.91,81.84,66.90,62.78,56.60,50.77,42.11,40.67,40.09,39.20,37.69,37.48,32.82,32.19,32.01,31.27,30.31,29.10,28.82,21.09,19.33,17.52,15.46,15.40, 14.78. TOF-HRMS: m/e (506.7179), formula: c₃₂H₄₆O₃N₂Compound 66 in table 1.

Compound 64(R ═ 3-pyridazinyl), compound 65(R ═ 4-pyridazinyl), compound 67(R ═ 5-methyl-3-pyridazinyl), compound 68(R ═ 3-methyl-4-pyridazinyl) were each prepared according to a similar manner to example 22 using an appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 23

Weighing diosgenin 4.2g, dissolving in tetrahydrofuran 100mL in a 500mL three-neck flask, adding triethylamine 10mL and 4-bromomethylpyridazine 2.2g, stirring, heating to 66 deg.C (heating reflux), reacting for 0.5h under heat preservation, detecting total reaction of diosgenin by TLC, heating to room temperature, extracting with ethyl acetate for 3 times (100mL × 3), evaporating under reduced pressure to liquid volume 40mL, cooling to 5 deg.C, and condensingCrystallization overnight, filtration, TLC tracing reaction and product separation and purification process, solid at 60 degrees C drying for 4h, get pale yellow powder product 3.2 g. Melting point of the target product: 184.0-186.0 ℃ of the total weight of the mixture,¹³C NMR(101MHz,CDCl₃) δ 149.72,149.70,140.78,135.55,129.03,121.80,119.59,87.14,82.90,72.83,66.89,62.77,56.59,50.76,42.10,40.66,40.08,39.19,37.68,37.47,32.81,32.18,32.00,31.26,30.30,29.09,28.81,21.08,19.32,17.51,15.45, 14.77. TOF-HRMS: m/e (506.7178), formula: c₃₂H₄₆O₃N₂Compound 70 in table 1.

Compound 69(R ═ 3-pyridazinylmethyl) can be prepared by a method similar to that of example 23, using the appropriate halogenated nitrogen-containing heterocyclic group (halogen may be Cl or Br).

Example 24

Weighing 4.2g of diosgenin, dissolving in 100mL of dichloromethane in a 500mL three-neck flask, adding 10mL of diethylamine and 2.2g of 2-bromomethylpyrazine, stirring, reacting at 0 ℃ under heat preservation for 48h, detecting all reactions of the diosgenin by HPLC, evaporating the dichloromethane until the volume of the liquid is reduced to 1/4, cooling to 5 ℃ for crystallization overnight, filtering, tracing the separation and purification processes of the reaction and the product by HPLC, and drying the solid at 60 ℃ for 4h to obtain 3.1g of a yellow powdery product. Melting point of the target product: 182.5-184.4 ℃ of the total weight of the mixture,¹³C NMR(CDCl₃100MHz) delta 153.75,144.81,143.85,142.43,140.86,121.88,119.67,87.10,82.98,71.82,66.97,62.85,56.67,50.84,42.18,40.74,40.16,39.27,37.75,37.55,32.89,32.26,32.08,31.34,30.38,29.17,28.89,21.16,19.40,17.59,15.83, 14.85. TOF-HRMS: m/e (506.7175), formula: c₃₂H₄₆O₃N₂Compound 73 in table 1.

Compound 71(R ═ 2-pyrazinyl) and compound 72(R ═ 3-methyl-2-pyrazinyl) can be prepared according to a similar procedure to that of example 24, using the appropriate halogenated nitrogen-containing heterocyclic group (halogen can be Cl or Br), respectively.

Example 25

Screening experiment for inhibitory activity on proliferation of bile duct cancer cell QBC-939 and human pancreatic cancer cell PANC-1

Collecting bile duct cancer cell QBC-939 and human pancreatic cancer cell PANC-1 with good growth state in logarithmic growth phase at 2 × 10³Per 0.1 mL/well, plated in 96-well plates, each set of 5 parallel wells, 5% CO at 37 ℃²Culturing in incubator, culturing in culture solution containing 10% calf serum for 24 hr, and respectively changing to compound 1-73[ concentration of 0.2ug/mL ] in Table 1]The culture medium is continuously cultured, the culture medium is added into a negative group, 0.2ug/mL 5-fluorouracil (national drug group chemical reagent Co., Ltd.) is added into a control group 1, and 0.2ug/mL diosgenin (Aladdin chemical reagent Co., Ltd.) is added into a control group 2. After 24h of culture, adding MTT20uL (5mg/mL) for further culture for 4h, discarding the supernatant, adding 100uL of dimethyl sulfoxide (DMSO), shaking for 10min, after the precipitate is completely dissolved, adjusting the wavelength to 570nm on an enzyme-linked detection analyzer, adjusting the blank to zero, determining the OD value of each well, and repeating the steps for 3 times. The inhibition rate is (1-drug-added OD value/negative group OD value) × 100%

The synthesized typical compounds 1 to 73 and 5-fluorouracil have the inhibitory activity on the proliferation of bile duct cancer cells QBC-939 and human pancreatic cancer cells PANC-1, and the experimental results are shown in Table 2.

(R groups are shown in Table 2)

TABLE 2 inhibition of proliferation of bile duct cancer QBC-939 and human pancreatic cancer PANC-1 cells

Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A compound having the structure shown in formula (I):

wherein R is an unsubstituted or substituted nitrogen-containing heterocyclic group; the nitrogen heterocyclic group is a five-membered or six-membered nitrogen-containing aliphatic heterocyclic ring or a nitrogen-containing aromatic heterocyclic ring; the substituent is selected from alkyl, halogenated alkyl, carboxyl and halogen;

or a pharmaceutically acceptable salt of the compound of formula (I).

2. A compound according to claim 1, wherein the nitrogen-containing heterocycle is selected from pyrrole, thiazole, imidazole, pyrazole, oxazole, isoxazole, pyridine, pyrimidine, pyridazine, pyrazine and piperidine.

3. A compound according to claim 1 or 2, characterised in that it isThe substituents being selected from C_1-3Alkyl, trifluoromethyl and carboxyl;

preferably, the substitution is mono-or di-substituted.

4. A compound according to claim 1 or 2, wherein R is selected from the group consisting of: 2-pyrrolyl, 3-pyrrolyl, 2-methyl-4-pyrrolyl, 2-imidazolyl, 4-methyl-2-imidazolyl, 2-methyl-4-imidazolyl, 3-pyrazolyl, 4-pyrazolyl, 5-methyl-3-pyrazolyl, 3-methyl-4-pyrazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 2-oxazolylmethyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 4-isoxazolyl, 3, 5-dimethylisoxazolyl-4-methyl, 2-imidazolyl, 4-pyrazolyl, 4-thiazolyl, 5-thiazolyl, 2-thiazolyl, 4-oxazolyl, 3-dimethylisoxazolyl-4-methyl, 2-oxazolyl, 3-dimethylisoxazolyl, 4-methyl, and mixtures thereof, 2-pyridyl group, 3-pyridyl group, 4-pyridyl group, 3-methyl-2-pyridyl group, 4-methyl-2-pyridyl group, 5-methyl-2-pyridyl group, 2-methyl-3-pyridyl group, 4-methyl-3-pyridyl group, 5-methyl-3-pyridyl group, 2-methyl-4-pyridyl group, 3-trifluoromethyl-2-pyridyl group, 4-trifluoromethyl-2-pyridyl group, 5-trifluoromethyl-2-pyridyl group, 2-trifluoromethyl-3-pyridyl group, 4-trifluoromethyl-3-pyridyl group, 5-trifluoromethyl-3-pyridyl group, 3-trifluoromethyl-pyridyl group, 3-pyridyl group, 4-trifluoromethyl-2-pyridyl group, 3-pyridyl group, 4-trifluoromethyl-pyridyl group, 3-, 2-trifluoromethyl-4-pyridyl group, 3-carboxy-2-pyridyl group, 4-carboxy-2-pyridyl group, 5-carboxy-2-pyridyl group, 2-carboxy-3-pyridyl group, 4-carboxy-3-pyridyl group, 5-carboxy-3-pyridyl group, 2-carboxy-4-pyridyl group, 3-pyridylmethyl group, 4-pyridylmethyl group, 2-pyrimidyl group, 4-pyrimidyl group, 5-pyrimidyl group, 4-methyl-2-pyrimidyl group, 5-methyl-2-pyrimidyl group, 2-methyl-4-pyrimidyl group, 3-carboxy-2-pyridyl group, 5-carboxy-3-pyridyl group, 2-methyl-5-pyrimidinyl, 2-pyrimidylmethyl, 4-pyrimidylmethyl, 5-pyrimidylmethyl, 3-pyridazinyl, 4-methyl-3-pyridazinyl, 5-methyl-3-pyridazinyl, 3-methyl-4-pyridazinyl, 3-pyridazinylmethyl, 4-pyridazinylmethyl, 2-pyrazinyl, 3-methyl-2-pyrazinyl and 2-pyrazinylmethyl.

5. A preparation method of a compound shown in a formula (I) is characterized by comprising the steps of taking diosgenin as an initial reactant to react with RX to prepare the compound shown in the formula (I);

wherein R is as defined in any one of claims 1 to 4 and X is halogen.

6. A preparation method according to claim 5, which comprises dissolving diosgenin in solvent, adding RX and alkaline substance, reacting, and keeping the solution alkaline during the reaction to obtain compound of formula (I);

preferably, the reaction temperature is-20-300 ℃, and the reaction time is 0.1-72 h;

the solvent is selected from one or more of water, methanol, ethanol, propanol, butanol, pentanol, glycerol, dichloromethane, trichloromethane, carbon tetrachloride, benzene, toluene, xylene, 1, 4-dioxane, 1, 2-dichloroethane, tetrahydrofuran, N-dimethylformamide, ethyl acetate and dimethyl sulfoxide;

preferably, the amount of the solvent is such that each mole of diosgenin is dissolved in 5-100L of solvent;

preferably, the alkaline substance is selected from gamma-Al₂O₃-Na、γ-Al₂O₃-K、γ-Al₂O₃-NaOH-Na、γ-Al₂O₃-NaOH-K、γ-Al₂O₃-KOH-Na、γ-Al₂O₃-one or more of KOH-K, sodium methoxide, sodium ethoxide, sodium propoxide, sodium butoxide, sodium amide, sodium metal, sodium hydride, sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium oxide, potassium oxide, calcium oxide, strontium carbonate, potassium carbonate, sodium carbonate, potassium phosphate, ammonia, ethylamine, diethylamine, triethylamine, methylamine, dimethylamine, trimethylamine, pyridine, and piperidine;

preferably, the molar ratio of the diosgenin to the RX to the alkaline substance is 1: (0.1-10): (0.01-10), preferably 1 (1.0-1.2): (0.6-1.0).

7. The method as claimed in claim 5 or 6, further comprising detecting the end point of the reaction by chromatography, standing the reaction product at room temperature after the reaction is finished, and preparing the compound of formula (I) by separation and purification;

preferably, the separation and purification method is selected from one or more of filtration, resin treatment, water washing, distillation, crystallization, extraction, activated carbon treatment, molecular sieve treatment and chromatography.

8. A composition comprising a compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof.

9. A pharmaceutical formulation comprising a compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof or a composition according to claim 8, together with at least one pharmaceutical carrier or adjuvant.

10. Use of a compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, or a composition of claim 8, or a pharmaceutical formulation of claim 9, in the manufacture of an anti-cancer medicament;

preferably, the cancer is selected from pancreatic cancer and cholangiocarcinoma.