CN118085002A

CN118085002A - Process for preparing cholic acid derivatives

Info

Publication number: CN118085002A
Application number: CN202410209983.3A
Authority: CN
Inventors: 仇文卫; 宋保亮; 江倩茹; 江世友; 史熊杰; 李晨晨
Original assignee: Wuhan University WHU; East China Normal University
Current assignee: Wuhan University WHU; East China Normal University
Priority date: 2020-04-28
Filing date: 2020-04-28
Publication date: 2024-05-28
Also published as: CN113563404A; CN113563404B; CN118085007A

Abstract

The invention discloses a cholic acid derivative shown in a formula (I) and a preparation method thereof, and the cholic acid derivative is prepared by TBS protection, 4-dimethyl, deprotection, oxidation, witting reaction, reduction, hydrolysis, acylation, deprotection and other reactions. The invention also provides application of the cholic acid derivative in preparing medicines for preventing and/or treating hypercholesterolemia, hypertriglyceridemia, atherosclerosis and nonalcoholic steatohepatitis, the cholic acid derivative not only can effectively promote the degradation of hydroxymethylglutaryl-CoA reductase (3-hydroxy-3-methyl-glutaryl-coenzymeAreductase, HMGCR), but also can reduce the increase of HMGCR protein caused by statin medicines, thereby reducing endogenous cholesterol level, providing a beneficial reference for the research and development of cholesterol-lowering new medicines, and having good application prospects.

Description

Process for preparing cholic acid derivatives

Technical Field

The invention belongs to the technical field of medicines and preparation and application thereof, and particularly relates to a cholic acid derivative, a preparation method thereof and application thereof in the aspects of atherosclerosis resistance and nonalcoholic steatohepatitis resistance.

Background

Cardiovascular disease is a type of disease with a very high current incidence. The excessive cholesterol content in blood, especially the low density lipoprotein, is one of the most important factors. The currently prevailing drugs for lowering high cholesterol are mainly statin drugs including: atorvastatin (atovastatin), simvastatin (Simvastatin), lovastatin (Lovastatin), pravastatin (PRAVASTATIN), fluvastatin (Fluvastatin), rosuvastatin (Rosuvastatin) and the like. The action mechanism of the statin drugs mainly simulates HMG-CoA, so that the statin drugs can competitively bind to HMG-CoA reductase (HMGCR) to inhibit the conversion of HMG-CoA into mevalonate, and the biosynthesis of cholesterol in vivo is limited.

The safety of statins is becoming more and more of a concern, and the most serious common side effects are liver damage and muscle toxicity (muscle pain, myositis, rhabdomyolysis, etc.), other manifestations including gastrointestinal discomfort, headache, sleep disorders, peripheral neuropathy, etc. Especially, after long-term administration of statin, HMGCR protein is compelled to be increased, so that the patient has to suppress intracellular increased HMGCR by increasing the drug dosage, which seriously impairs the therapeutic effect of statin and increases side effects. Although this disadvantage of statins is recognized from the beginning at the time of clinical use, there is no way to date to prevent statin-induced HMGCR accumulation and its side effects. Aiming at HMGCR protein ubiquitination degradation, a first generation of lithocholic acid series derivatives are designed and synthesized for the first time in a laboratory, and a plurality of active small molecular compounds with novel structures are screened and obtained from the derivatives. Of these, the representative compound HMG499 has HMGCR protein ubiquitination promoting degradation activity of EC ₅₀ =0.41 μm. The compound not only can obviously promote the ubiquitination degradation of HMGCR protein at the cellular level, but also can reduce the accumulation of HMGCR protein induced by statin, and can effectively inhibit the generation of cholesterol; but also has obvious lipid-lowering and cholesterol-lowering effects at animal level. The study was published in 2018, 12, and published in the international journal of authority, nature communication (Nature Communications,2018,9,5138), and through subsequent studies we found that the first generation of compounds involved in the paper, while significantly promoting ubiquitination degradation of HMGCR protein, reducing statin-induced accumulation of HMGCR protein, and effectively inhibiting cholesterol synthesis in mice, reducing atherosclerotic plaque production, also resulted in intracellular cholesterol accumulation (as shown in FIG. 1).

Cholesterol accumulation affects the survival of nerve cells, causing severe neurodegenerative diseases such as niemann-pick type C; in addition, intracellular cholesterol accumulation can lead to excessive accumulation of lipid material in the liver, kidneys, spleen and brain, which can lead to lesions and possibly lethality in these organs. How to solve the problem of cholesterol accumulation caused by the compounds becomes a key in the development process of new drugs of the compounds.

Disclosure of Invention

On the basis of retaining the excellent activity of the first generation lithocholic acid derivatives, the invention further modifies the structure of the compounds, obtains a series of compounds which do not generate cholesterol accumulation at all, and has important significance for the research and development of the new drugs.

The invention provides a cholic acid derivative, which has a structure shown in a formula (I):

Wherein n is selected from natural numbers,

R ₁ is selected from various heterocycles and substituted heterocycles.

R ₂ is selected from hydrogen, hydroxy, carbonyl, halogen, alkyl.

R ₃ is selected from alcohol, carboxyl, amide.

The C5 and C6 connecting bonds are single bonds or double bonds.

Preferably, the method comprises the steps of,

The natural number n is selected from 0,1,2,3,4.

R ₁ includes a pyrazine ring, a pyrazole ring, a substituted thiazole ring, a1, 2, 3-oxadiazole ring, a 2-aminopyrimidine ring, an isoxazole ring and a substituted isoxazole ring;

The pyrazine ring is

The pyrazole ring is

The substituted pyrazole ring includes N-acetyl pyrazoleN-propionyl pyrazoleN-butyrylpyrazoleN-isobutyrylpyrazoleN-methacryloylpyrazole1-Phenylpyrazole ring1- (2-Fluoro) phenylpyrazole ring

Preferably, the substituted pyrazole ring is N-acetyl pyrazoleN-isobutyrylpyrazole

Further preferably, the substituted pyrazole ring is N-acetyl pyrazole

The substituted thiazole ring includes a 2-methylthiazole ring2-Aminothiazole ring

The 1,2, 3-oxadiazole ring is

The 2-aminopyrimidine ring is

The isoxazole ring is

The substituted isoxazole ring is

The hydroxy group of R ₂ is in alpha or beta configuration, preferably beta configurationHydrogen is

The R ₃ is selected from tertiary alcohol, carboxyl and amide;

The tertiary alcohol comprises diethyl tertiary alcohol Diallyl tertiary alcoholDimethyl tertiary alcoholDi-n-propyl tertiary alcoholIsopropyl tertiary alcoholDi-n-butyl tertiary alcoholDiisopropyltertiary alcoholDipropargyl tertiary alcoholDiphenyl tertiary alcoholDibenzyl tertiary alcohol

Preferably, the tertiary alcohol comprises diethyl tertiary alcoholDiallyl tertiary alcohol

The carboxyl group is

The amide comprises formamideAcetamideN-propionamide2-MethylbenzamidePara-toluamide5-Nitrothiazole amideN-3-pyridinecarboxamidesPiperidine amideMorpholinamideN-Boc-4-piperidinamideN-acetyl-4-piperidinamideHexahydropyran-4-amideMorpholine-2-ethylamide

The invention also provides a cholic acid derivative, which has a structure shown in a formula Q27:

The invention also provides a cholic acid derivative, which has a structure shown in a formula Q63:

The invention also provides a preparation method of the cholic acid derivative, which takes the cholic acid derivative shown in the formula Q1 as a starting raw material, and the compound Q7 is obtained through TBS protection, 4-dimethyl, TBS protection removal, oxidation, witting reaction and reduction; then, taking the compound Q5 as a starting material, and oxidizing and esterifying to obtain a compound Q9; or taking the compound Q4 as a starting material, and respectively obtaining a compound Q13 through iodination, cyano substitution, hydrolysis and esterification; the preparation method is shown in a route (1):

specifically, the method comprises the steps of:

(a) TBS protection reaction

And (3) dissolving the compound Q1 in an organic solvent, adding a reagent and alkali for TBS protection at a low temperature, and carrying out TBS protection reaction to obtain a compound Q2.

In the step (a), the organic solvent is selected from any one or more of N, N-dimethylformamide, N-dimethylacetamide, methylene dichloride, chloroform, carbon tetrachloride and the like; preferably, it is N, N-dimethylformamide.

In the step (a), the base is selected from any one or more of triethylamine, imidazole, diisopropylethylamine and the like; preferably imidazole.

In the step (a), the molar ratio of the compound Q1 to the alkali is 1 (2-20); preferably 1:8.

In the step (a), the reagent used for TBS protection is selected from any one or more of tert-butyldimethylsilyl chloride (TBSCl), tert-butyldimethylsilyl triflate (TBSOTf) and the like; preferably TBSCl.

In the step (a), the molar ratio of the compound Q1 to the reagent used for TBS protection is selected from 1 (1) to 10); preferably 1:4.

In the step (a), the low temperature is-10 ℃; preferably at 0 ℃.

In the step (a), the temperature of the TBS protection reaction is 25-70 ℃; preferably 25 ℃.

In the step (a), the time of the TBS protection reaction is 2-12 h; preferably 5h.

(B) 4, 4-Dimethylation reaction

Dissolving the compound Q2 in an organic solvent, adding alkali and a methylation reagent, and carrying out methylation reaction to obtain a compound Q3.

In the step (b), the organic solvent is selected from any one or more of t-BuOH, benzene, carbon tetrachloride, tetrahydrofuran and the like; preferably, t-BuOH.

In the step (b), the base is selected from any one or more of t-BuOK, KOC (Et) Me ₂ and the like, preferably t-BuOK.

In the step (b), the methylating agent is any one or more of CH ₃Cl、CH₃Br、CH₃ I and the like; preferably CH ₃ I.

In the step (b), the molar ratio of the compound Q2 to the alkali is 1 (2-8); preferably 1:4.

In the step (b), the molar ratio of the compound Q2 to the methylating agent is 1 (2-10); preferably 1:8.

In the step (b), the optimal adding method of the methylation reagent is slow dripping.

In step (b), the base is added in portions.

In step (b), the temperature of the reaction during the addition of the methylating agent is 0 ℃.

In step (b), the temperature of the methylation reaction is 0 to 60 ℃, preferably 25 ℃.

In the step (b), the methylation reaction time is 2-12 h; preferably 4h.

(C) TBS removal protection reaction

And (3) dissolving the compound Q3 in an organic solvent, adding an acid required for deprotection, and reacting to obtain the compound Q4.

In the step (c), the organic solvent is any one or more of diethyl ether, tetrahydrofuran, ethyl acetate, methanol, ethanol and the like; preferably diethyl ether.

In the step (c), the acid is any one or more of p-toluenesulfonic acid, hydrochloric acid gas, sulfuric acid, hydrochloric acid solution and the like; preferably, it is hydrochloric acid gas.

In the step (c), the time for removing TBS protection reaction is 1-5 h; preferably 2h.

In the step (c), the temperature of the TBS removal protection reaction is 25-60 ℃; preferably 25 DEG C

(D) Oxidation reaction

And (3) dissolving the compound Q4 in an organic solvent, adding an oxidant, and reacting to obtain the compound Q5.

In the step (d), the organic solvent is selected from any one or more of toluene, tetrahydrofuran, dichloromethane and the like; preferably, it is dichloromethane.

In the step (d), the oxidant is any one or more of PDC, DDQ, PCC, IBX and the like; preferably PDC.

In the step (d), the molar ratio of the compound Q4 to the oxidant is 1 (1-3); preferably 1:1.5.

In the step (d), the temperature of the oxidation reaction is 20-35 ℃; preferably 25 ℃.

In the step (d), the time of the oxidation reaction is 7-10 hours; preferably 8h.

(E) Witting reaction

Dissolving alkali in an organic solvent, respectively adding a Witting reagent and a compound Q5, and reacting to obtain a compound Q6.

In the step (e), the organic solvent is selected from any one or more of tetrahydrofuran, toluene, petroleum ether, benzene and the like; preferably tetrahydrofuran.

In the step (e), the alkali is one or more of sodium methoxide, sodium ethoxide and sodium hydride (60 percent); preferably sodium hydride (60%).

In step (e), the Witting reagent is preferably triethyl phosphorylacetate.

In the step (e), the molar ratio of the compound Q5 to the alkali and the Witting reagent is 1 (3-8); preferably 1:4.5:5.

In the step (e), the adding mode of the Witting reagent is dripping, and the dripping time is 5-30 min; preferably 10min.

In the step (e), the temperature of the Witting reaction is 25-75 ℃; preferably 25 ℃.

In the step (e), the time of the Witting reaction is 1-3 h; preferably 1h.

(F) Reduction reaction

Dissolving a compound Q6 in an organic solvent, adding a reducing agent to obtain a compound intermediate, and then adding an oxidizing agent to oxidize to obtain a compound Q7.

In step (f), the organic solvent of the reduction reaction is preferably dried anhydrous methanol.

In step (f), the reducing agent used in the reduction reaction is preferably magnesium turnings.

In the step (f), the molar ratio of the compound Q6 to the reducing agent is 1 (2-10); preferably 1:10.

In the step (f), the temperature of the reduction reaction is 25-60 ℃; preferably 25 ℃.

In the step (f), the reaction time for obtaining the intermediate is 3-8 hours; preferably 5h.

In the step (f), the organic solvent for the oxidation reaction is selected from any one or more of toluene, tetrahydrofuran, DMSO and the like; preferably, it is a solvent mixture of DMSO and toluene.

In step (f), the oxidizing agent functions to oxidize a small amount of reduced hydroxyl groups at the C-3 position.

In step (f), the oxidizing agent is selected from any one or more of IBX, PDC, PCC, DDQ and the like; preferably IBX.

In the step (f), the molar ratio of the compound Q6 to the oxidant is 1 (0.5-3); preferably 1:1.

In the step (f), the temperature of the oxidation reaction is 25-60 ℃; preferably 25 ℃.

In the step (f), the time of the oxidation reaction is 2-8 hours; preferably, 4h.

(G) Oxidation reaction

Dissolving the compound Q5 in an organic solvent, adding an oxidant at a low temperature, and reacting to obtain the compound Q8.

In the step (g), the organic solvent for the oxidation reaction is selected from any one or more of pyridine, chloroform, dichloroethane, 1, 2-dichloropropane and the like; preferably, pyridine.

In the step (g), the temperature of the reaction liquid is 0-10 ℃ when the oxidant is added; preferably at 0 ℃.

In the step (g), the oxidant is selected from any one or more of tetrabutylammonium permanganate, potassium dichromate, O ₃ and the like; preferably tetrabutylammonium permanganate.

In step (g), the molar ratio of Q5 to oxidant is 1: (1-4); preferably, it is 1:2.

In the step (g), the temperature of the oxidation reaction is 0-60 ℃; preferably 25 ℃.

In the step (g), the time of the oxidation reaction is 1-2 h; preferably 1h.

(H) Esterification reaction

Dissolving the compound Q8 in an organic solvent, and adding acid to perform esterification reaction to obtain a compound Q9.

In the step (h), the organic solvent is selected from any one or more of methanol, tetrahydrofuran, a mixed solvent of methanol and the like; preferably, methanol.

In the step (h), the acid in the esterification reaction is selected from any one or more of concentrated sulfuric acid, p-toluenesulfonic acid, thionyl chloride and the like; preferably concentrated sulfuric acid.

In the step (h), the molar ratio of the compound Q8 to the acid is 1 (0.5-2); preferably 1:0.7.

In the step (h), the temperature of the esterification reaction is 25-70 ℃; preferably 70 ℃.

In the step (h), the time of the esterification reaction is 2-12 h; preferably 2h.

(I) Iodination reaction

Dissolving an iodination reagent, a catalyst and an acid binding agent in an organic solvent, stirring for a period of time, and then adding a compound Q4 to react to obtain a compound Q10.

In the step (i), the organic solvent is selected from one or more of toluene, methylene dichloride, benzene and the like; preferably toluene.

In the step (I), the iodination reagent is selected from any one or more of I ₂, N-iodinated succinimide and the like; preferably, I ₂.

In the step (i), the catalyst and the acid binding agent in the iodination reaction are PPh ₃ and imidazole respectively.

In the step (i), the molar ratio of the compound Q4 to the iodination reagent, the catalyst and the acid binding agent is 1 (3-8): 6-16; preferably 1:7:7.3:14.

In the step (i), the reaction temperature before the iodination reaction is added into the compound Q4 is 25-60 ℃; preferably 25 ℃.

In the step (i), the reaction time before the iodination reaction is added into the compound Q4 is 0.5 to 3 hours; preferably 1h.

In the step (i), the reaction temperature is 25-60 ℃ after the iodination reaction is carried out and the compound Q4 is added; preferably 25 ℃.

In the step (i), the reaction time after the iodination reaction is added with the compound Q4 is 1-2 h; preferably 2h.

(J) Cyano substitution reaction

Dissolving the compound Q10 in an organic solvent, adding alkali and a cyanation reagent, and reacting to obtain the compound Q11.

In the step (j), the organic solvent is selected from one or more of DMF, tetrahydrofuran, DMSO, toluene and the like; preferably, DMF.

In step (j), the base is selected from one or more of sodium hydroxide, potassium fluoride, sodium fluoride, potassium acetate, sodium acetate, and the like; preferably potassium fluoride.

In the step (j), the cyanating reagent is selected from one or more of sodium cyanide, trimethylsilyl cyanide and the like; preferably trimethylcyanogen.

In the step (j), the molar ratio of the compound Q10 to the alkali and the cyanating reagent is 1 (2-20): 2-10; preferably 1:16:8.

In the step (j), the temperature of the cyanogen extraction reaction is 40-80 ℃; preferably 50 ℃.

In the step (j), the time of the cyanogen extraction reaction is 2-12 h; preferably 4h.

(K) Hydrolysis reaction

Dissolving the compound Q11 in an organic solvent, adding alkali, and hydrolyzing to obtain a compound Q12.

In the step (k), the organic solvent is selected from any one or more of methanol, tetrahydrofuran, ethanol, a mixed solvent of water and methanol, and the like; preferably, a solvent mixture of water and ethanol.

In the step (k), the volume ratio of the water and the ethanol mixed solvent is 1 (1-3); preferably 1:3.

In the step (k), the alkali is selected from any one or more of lithium hydroxide, sodium hydroxide, potassium carbonate and the like; preferably sodium hydroxide.

In the step (k), the molar ratio of the compound Q11 to the alkali is 1 (1-10); preferably 1:10.

In the step (k), the temperature of the hydrolysis reaction is 70-120 ℃; preferably at 100 ℃.

In the step (k), the time of the hydrolysis reaction is 24-72 h; preferably 72h.

(L) Esterification reaction

The synthesis of compound Q13 is similar to the esterification process of compound Q9 in scheme (1).

The invention also provides a preparation method of the cholic acid derivative, which takes the compound Q7 as a starting material, and the compound Q19 is obtained through glycol protection, reduction, iodo, cyano substitution, hydrolysis and esterification, and the preparation method is shown in a route (2):

specifically, the method comprises the steps of:

(a) Glycol protection reaction

And (3) dissolving the compound Q7 in an organic solvent, adding a catalyst, a dehydrating agent and ethylene glycol, and reacting to obtain a compound Q14.

In the step (a), the organic solvent is selected from tetrahydrofuran, a mixed solution of tetrahydrofuran and ethylene glycol; preferably, the solution is a mixed solution of tetrahydrofuran and ethylene glycol.

In the step (a), the volume ratio of tetrahydrofuran to glycol is 3 (1-3); preferably 3:1.

In step (a), the catalyst is preferably p-toluene sulfonic acid.

In step (a), the dehydrating agent is selected from triethyl orthoformate, trimethyl orthoformate; preferably, it is triethyl orthoformate.

In the step (a), the molar ratio of the compound Q7 to the catalyst to the dehydrating agent is 1 (0.2-0.5) (5-10); preferably 1:0.2:5.

In the step (a), the time of the glycol protection reaction is 2-12 h; preferably 3h.

In the step (a), the temperature of the glycol protection reaction is 25-60 ℃; preferably 25 ℃.

(B) Reduction reaction

Dissolving the compound Q14 in an organic solvent, adding a reducing agent, and reacting to obtain the compound Q15.

In the step (b), the organic solvent is selected from any one or more of tetrahydrofuran, diethyl ether, methanol, ethanol and the like; preferably tetrahydrofuran.

In the step (b), the reducing agent is any one or more of LiAlH ₄、NaBH₄, red aluminum, diisobutyl aluminum hydride and the like; preferably LiAlH ₄.

In step (b), the reducing agent is preferably added in portions.

In the step (b), the molar ratio of the compound Q14 to the reducing agent is 1 (2-20); preferably 2:10.

In the step (b), the temperature of the reduction reaction is-10 ℃; preferably at 0 ℃.

In the step (b), the time of the reduction reaction is 1-5 h; preferably 2h.

(C) Iodination reaction

The synthesis of compound Q16 is similar to the iodination method of compound Q10 in scheme (1).

(D) Cyano substitution reaction

The synthesis of compound Q17 is similar to the substitution method of compound Q11 in scheme (1).

(E) Hydrolysis reaction

The synthesis of compound Q18 is similar to the hydrolysis method of compound Q12 in scheme (1).

(F) Esterification reaction

The synthesis of compound Q19 is similar to the esterification process of compound Q9 in scheme (1).

The invention also provides a preparation method of the cholic acid derivative, which takes the compound Q15 as a raw material, and the compound Q23 is obtained through oxidation, witting reaction, glycol removal protection and reduction, wherein the preparation method is shown in a route (3):

specifically, the method comprises the steps of:

(a) Oxidation reaction

And dissolving the compound Q15 in an organic solvent, adding an oxidant, and reacting to obtain the compound Q20.

In the step (a), the organic solvent is selected from any one or more of DMSO, toluene, tetrahydrofuran, a mixed solution of DMSO and toluene and the like; preferably, it is a mixed solution of DMSO and toluene.

In the step (a), the oxidant is selected from any one or more of IBX, DDQ, PCC, PDC and the like; preferably IBX.

In the step (a), the molar ratio of the compound Q15 to the oxidant is 1 (1-3); preferably 1:1.5.

In the step (a), the temperature of the oxidation reaction is 20-35 ℃; preferably 25 ℃.

In the step (a), the time of the oxidation reaction is 7-10 hours; preferably 8h.

(B) Witting reaction

The synthesis of compound Q21 is similar to the synthetic reaction method of compound Q6 in scheme (1).

(C) Deprotection reaction of ethylene glycol

Dissolving the compound Q21 in an organic solvent, adding acid, and reacting to obtain the compound Q22.

In the step (c), the organic solvent is selected from one or more of methanol, ethanol, tetrahydrofuran, a mixed solvent of methanol and water, a mixed solvent of ethanol and water, a mixed solvent of tetrahydrofuran and water, and the like; preferably, a mixed solvent of methanol and water.

In step (c), the acid is selected from one or more of dilute sulfuric acid, 2M dilute hydrochloric acid, hydrochloric acid gas, p-toluenesulfonic acid and the like; preferably, it is a 2M dilute aqueous hydrochloric acid solution.

In the step (c), the volume ratio of the reaction solvent to the 2M diluted hydrochloric acid is 5 (1-2); preferably 5:1.

In the step (c), the reaction time is 1-5 h; preferably 2h.

In the step (c), the temperature of the reaction is 0-60 ℃; preferably 25 ℃.

(D) Reduction reaction

The synthesis of compound Q23 is similar to the reduction reaction method of compound Q7 in scheme (1).

The invention also provides a preparation method of the cholic acid derivative, which takes the compound Q7 as a starting material, and the compound Q27 is obtained through condensation, addition cyclization, hydrolysis and acetylation; then condensing the compound Q27 serving as a starting material with an amine compound to obtain cholic acid derivatives shown in the formulas Q28-Q39; taking a compound Q25 as a starting material, and carrying out Grignard reaction with a Grignard reagent to obtain a cholic acid derivative Q56; the preparation method is shown in a route (4):

specifically, the method comprises the steps of:

(a) Condensation reaction

And dissolving the compound Q7 in an organic solvent, adding a condensing agent, and reacting to obtain the compound Q24.

In step (a), the condensation reaction is preferably carried out under nitrogen protection.

In the step (a), the organic solvent is selected from tetrahydrofuran, ethyl formate and a mixed solvent of tetrahydrofuran and ethyl formate; preferably, it is ethyl formate.

In the step (a), the condensing agent is selected from one or more of sodium hydride (60%), sodium methoxide, sodium ethoxide and the like; preferably sodium hydride (60%).

In the step (a), the molar ratio of the compound Q7 to sodium hydride is 1 (5-10); preferably 1:10.

In the step (a), the temperature of the condensation reaction is 25-45 ℃; preferably 25 ℃.

In the step (a), the time of the condensation reaction is 20-60 min; preferably 20min.

(B) Addition cyclization reaction

Dissolving the compound Q24 in an organic solvent, adding hydrazine hydrate and alkali, and reacting to obtain the compound Q25.

In the step (b), the organic solvent is selected from one or more of ethanol, ethanol-water mixed solvent and acetic acid; preferably acetic acid.

In the step (b), the alkali used in the addition cyclization reaction is selected from one or more of sodium acetate, potassium acetate, sodium ethoxide and potassium ethoxide; preferably potassium acetate.

In the step (b), the molar ratio of the addition cyclization reaction compound Q24 to the hydrazine hydrate to the alkali is 1 (1-3): 1-2; preferably 1:3:2.

In the step (b), the temperature of the addition cyclization reaction is 80-140 ℃; preferably 130 ℃.

In the step (b), the time of the addition cyclization reaction is 3-10 hours; preferably 5h.

(C) Hydrolysis reaction

Dissolving the compound Q25 in an organic solvent, adding alkali, and hydrolyzing to obtain a compound Q26.

In the step (c), the organic solvent is selected from any one or more of methanol, tetrahydrofuran, ethanol, a mixed solvent of water and methanol, and the like; preferably, a solvent mixture of water and ethanol.

In the step (c), the volume ratio of the water and the ethanol mixed solvent is 1 (1-3); preferably 1:3.

In the step (c), the alkali is selected from any one or more of lithium hydroxide, sodium hydroxide, potassium carbonate and the like; preferably lithium hydroxide.

In the step (c), the molar ratio of the compound Q25 to the alkali is 1 (1-40); preferably 1:30.

In the step (c), the temperature of the hydrolysis reaction is 10-70 ℃; preferably 25 ℃.

In the step (c), the time of the hydrolysis reaction is 1-12 h; preferably 8h.

(D) Acetylation reaction

Compound Q26 was dissolved in an organic solvent, and an acetylating agent and a catalyst were added to obtain compound Q27.

In the step (d), the organic solvent is selected from one or more of pyridine, triethylamine, diethylamine, diisopropylethylamine, tetrahydrofuran, DMF and the like; preferably tetrahydrofuran.

In the step (d), the acetylating reagent is selected from one or more of acetic anhydride, acetyl chloride and the like; preferably acetic anhydride.

In step (d), the acetylating reagent is preferably fed in portions.

In the step (d), the molar ratio of the Q26 to the acetylating reagent is 1 (1-5); preferably 1:3.

In step (d), the catalyst is selected from DMAP.

In step (d), the molar ratio of Q26 to DMAP is 1: (0.1 to 1); preferably, it is 1:0.2.

In the step (d), the temperature of the acetylation reaction is 0-100 ℃; preferably 60 ℃.

In the step (d), the time of the acetylation reaction is 1-5 h; preferably 2h.

(E) Acylation reaction

Compound Q27 was dissolved in an organic solvent and EDCI, HOBt, DMAP and the corresponding amine were added to give compounds Q28-Q39.

In the step (e), the organic solvent is selected from any one or more of DCM, diethyl ether, acetone, tetrahydrofuran, carbon tetrachloride, toluene, benzene, chloroform and the like; preferably, DCM.

In step (e), the EDCI, HOBt, DMAP acts to promote the amidation reaction.

In the step (e), the molar ratio of the compounds Q27, EDCI, HOBt, DMAP to the corresponding amine is 1 (1-2): 2-4): 1-2; preferably 1:2:2:4:2.

In the step (e), the temperature of the acylation reaction is 25-40 ℃; preferably 25 ℃.

In the step (e), the time of the acylation reaction is 6-12 h; preferably 12h.

(F) Format reaction

Compound Q25 was dissolved in an organic solvent and a formative reagent was added to give compound Q56.

In step (f), the reaction is preferably carried out under nitrogen.

In the step (f), the organic solvent in the reaction is any one or more of diethyl ether, anhydrous tetrahydrofuran, toluene, benzene and the like; preferably anhydrous tetrahydrofuran.

In the step (f), the format reagent is C ₂H₅ MgCl or CH ₂＝CHCH₂ MgCl; preferably, it is C ₂H₅ MgCl.

In the step (f), the temperature of the Grignard reaction is 0-60 ℃; preferably at 0 ℃.

In the step (f), the molar ratio of the compound Q25 to the formative reagent is 1 (10-30); preferably 1:20.

In the step (f), the Grignard reaction time is 0.5-5 h; preferably 2h.

The invention also provides a preparation method of the cholic acid derivative, which takes the compound Q36 as a starting material, and the cholic acid derivative Q43 is obtained through hydrolysis and condensation; the preparation method is shown in a route (5):

Dissolving a compound Q36 in an organic solvent, adding trifluoroacetic acid, and removing a Boc group to obtain an intermediate; the intermediate is then acetylated to give compound Q43.

Wherein, in the reaction of removing the Boc group, the organic solvent is selected from any one or more of dichloromethane, tetrahydrofuran, acetone, toluene and the like; preferably, it is dichloromethane.

Wherein the mol ratio of the reactant Q36 to the trifluoroacetic acid is 1 (2-3); preferably 1:3.

Wherein the Boc removal reaction time is 2-8 h; preferably 3h.

Wherein the temperature of the Boc removal reaction is 0-50 ℃; preferably 25 ℃.

Wherein the solvent used in the acetylation reaction is selected from one or more of pyridine, triethylamine, diethylamine, diisopropylethylamine, tetrahydrofuran, DMF and the like; preferably tetrahydrofuran.

Wherein the catalyst used in the acetylation reaction is selected from DMAP, and the molar ratio of the Q36 compound to the DMAP is 1: (0.1 to 1); preferably 1:0.2.

Wherein the acetylating agent is one or more of acetic anhydride, acetyl chloride and the like; preferably acetic anhydride.

Wherein, the optimal feeding mode of the acetylating reagent is fed in batches.

Wherein, the mol ratio of the Q36 and the acetylating reagent is 1 (2-6); preferably 1:4.

Wherein the temperature of the acetylation reaction is 0-100 ℃; preferably 60 ℃.

Wherein the time of the acetylation reaction is 1-5 h; preferably 2h.

The invention also provides a preparation method of the cholic acid derivative, which takes the compound Q24 as a starting material, and obtains the compounds Q45 and Q46 through addition cyclization, hydrolysis and acylation, or takes the compound Q44 as the starting material, and then carries out Grignard reaction with C ₂H₅ MgCl or CH ₂＝CHCH₂ MgCl respectively to obtain the cholic acid derivative shown as the formulas Q49 and Q50, wherein the preparation method is shown as a route (6):

specifically, the method comprises the steps of:

(a) Addition cyclization reaction

And dissolving the compound Q24 in an organic solvent, adding phenylhydrazine hydrochloride, and reacting to obtain the compound Q44.

In the step (a), the organic solvent is selected from one or more of ethanol, ethanol-water mixed solvent and acetic acid; preferably, it is a mixed solvent of ethanol and water.

In the step (a), the molar ratio of the reaction compound Q24 to phenylhydrazine hydrochloride is 1 (1-3); preferably 1:3.

In the step (a), the temperature of the reaction is 80-140 ℃; preferably, it is 100 ℃.

In the step (a), the reaction time is 3-10 h; preferably 3h.

(B) Hydrolysis reaction

The synthesis of compound Q45 is similar to the hydrolysis reaction method of Q26 in scheme (4).

(C) Acylation reaction

The synthesis of compound Q46 is similar to the acylation reaction of Q37 in scheme (4).

(D) Oxidation reaction

Compound Q44 is dissolved in an organic solvent, NHPI, acetic acid and an oxidant are added, and compound Q47 is obtained by reaction.

In step (d), the organic solvent is selected from one or more of acetone, DCM, acetic acid, etc.; preferably, acetone.

In the step (d), the oxidant is selected from one or more of sodium dichromate, potassium dichromate, chromium trioxide and the like; preferably, sodium dichromate is used.

In the step (d), the molar ratio of the compound Q44 to NHPI, acetic acid and oxidant is 1 (1-4): 0.2-2: (1-2); preferably 1:2:1:1.2.

In the step (d), the temperature of the oxidation reaction is 25-50 ℃; preferably 50 ℃.

In the step (d), the time of the oxidation reaction is 1-4 h; preferably 2h.

(E) Reduction reaction

And dissolving the compound Q47 in an organic solvent, adding a reducing agent, and reacting to obtain the compound Q48.

In the step (e), the solvent in the reduction reaction is selected from dichloromethane, methanol, a mixed solution of dichloromethane and methanol=1:1 or a mixed solution of dichloromethane and methanol=5:1; preferably, methanol.

In the step (e), the temperature of the reduction reaction is 25-50 ℃; preferably 25 ℃.

In the step (e), the reducing agent is selected from one or more of hydrogen, sodium borohydride, potassium borohydride and the like; preferably, sodium borohydride.

In the step (e), the molar ratio of the compound Q47 to the reducing agent is 1 (5-20); preferably 1:10.

In the step (e), the reaction time is 1-4 h; preferably 2h.

(F) Format reaction

The synthesis of compound Q49 was similar to the format reaction synthesis of Q56 in scheme (4).

The synthesis of compound Q50 is similar to the format reaction synthesis of Q56 in scheme (4).

The invention also provides a preparation method of the cholic acid derivative, which uses the compound Q26 as a starting material, and obtains the cholic acid derivative shown in the formulas Q51-Q55 through acylation and acid anhydride protection reaction, wherein the preparation method is shown in the route (7):

(a) Acylation reaction

The synthesis of compound Q51 is similar to the acylation reaction of Q37 in scheme (4).

(B) Acid anhydride protection reaction

The synthesis of compounds Q52-Q55 is similar to the acetylation reaction of Q27 in scheme (4).

The invention also provides a preparation method of the cholic acid derivative, which takes the compound Q7 as a starting material, and the compounds Q58, Q59 and Q61 are obtained through cyclization, hydrolysis, acylation and carbonyl reduction, and the preparation method is shown in a route (8):

(a) Cyclization reaction

And dissolving the compound Q7 in an organic solvent, adding sulfur and ethylenediamine, and reacting to obtain the compound Q57.

In step (a), the organic solvent is a morpholine.

In step (a), the sulfur is elemental sulfur powder.

In the step (a), the molar ratio of Q7 to sulfur and ethylenediamine is 1 (5-10): 5-10; preferably 1:10:10.

In the step (a), the temperature of the cyclization reaction is 100-150 ℃; preferably, it is 120 ℃.

In the step (a), the cyclization reaction time is 3-10 hours; preferably, 5h.

(B) Hydrolysis reaction

The synthesis of compound Q58 is analogous to the hydrolysis reaction method of Q26 in scheme (4).

(C) Acylation reaction

The synthesis of compound Q59 is similar to the acylation reaction method of Q37 in scheme (4).

(D) Oxidation reaction

The synthesis of compound Q60 is similar to the oxidation reaction method of Q47 in scheme (6).

(E) Reduction reaction

The synthesis of compound Q61 is similar to the reduction reaction method of Q48 in scheme (6).

The invention also provides a preparation method of the cholic acid derivative, which uses the compound Q7 as a starting material to obtain compounds Q64 and Q66 through bromination, cyclization and hydrolysis, wherein the preparation method is shown in a route (9):

(a) Bromination reaction

And dissolving the compound Q7 in an organic solvent, adding a brominating reagent, and reacting to obtain a compound Q62.

In the step (a), the organic solvent is selected from one or more of dichloromethane, methanol, CCl ₄, DMSO, DMF, benzene and the like; preferably, it is dichloromethane.

In step (a), the brominating reagent is one or more of pyridinium tribromide, NBS, br ₂, etc.; preferably, pyridinium tribromide.

In the step (a), the molar ratio of the compound Q7 to the brominating reagent is 1 (1-2); preferably 1:1.5.

In the step (a), the temperature of the bromination reaction is 0-30 ℃; preferably 25 ℃.

In the step (a), the bromination reaction time is 1-3 h; preferably 2h.

(B) Cyclization reaction

Dissolving the compound Q62 in an organic solvent, adding thiourea or thioacetamide, and reacting to obtain a compound Q63 or Q65.

In the step (b), the organic solvent is selected from one or more of ethanol, methanol and tetrahydrofuran; preferably, ethanol.

In the step (b), the molar ratio of the compound Q62 to the thiourea is 1 (1-3); preferably 1:2.

The molar ratio of the compound Q62 to the thioacetamide is 1 (1-3); preferably 1:2.

In the step (b), the temperature of the heterocyclic reaction is 80-120 ℃; preferably at 100 ℃.

In the step (b), the time of the heterocyclic reaction is 2-5 h; preferably 3h.

(C) Hydrolysis reaction

The synthesis of compounds Q64, Q66 is similar to the hydrolysis reaction method of Q26 in scheme (4).

The invention also provides a preparation method of the cholic acid derivative, which takes the compound Q7 as a starting material, and the cholic acid derivative Q68 is obtained through hydrolysis and acylation; or taking Q68 as a starting material, and obtaining cholic acid derivatives Q71 and Q72 through condensation, cyclization and acylation; or taking the compound Q68 as a starting material, and performing ester condensation and cyclization to obtain cholic acid derivatives Q74 or Q75 respectively; the preparation method is shown in a route (10):

(a) Hydrolysis reaction

The synthesis of compound Q67 is similar to the hydrolysis reaction method of Q26 in scheme (4).

(B) Acylation reaction

The synthesis of compound Q68 is similar to the acylation reaction of Q37 in scheme (4).

(C) Condensation reaction

Compound Q68 is dissolved in an organic solvent, alkali is added, after stirring is carried out for a period of time, isoamyl nitrite is dripped, and compound Q69 is obtained through reaction.

In step (c), the base is selected from one or more of potassium tert-butoxide, sodium methoxide, sodium formate, sodium ethoxide, potassium hydroxide and the like; preferably potassium tert-butoxide.

In the step (c), the organic solvent is selected from one or more of tertiary butanol, n-butanol, isopropanol, toluene, benzene, carbon tetrachloride and the like; preferably t-butanol.

In the step (c), the stirring temperature is 25-50 ℃ after the potassium tert-butoxide is added; preferably, 30 ℃;

in step (c), the stirring time after the addition of the potassium tert-butoxide is 0.5 to 3 hours, preferably 0.5 hours.

In the step (c), the isoamyl nitrite is slowly dropped.

In the step (c), the molar ratio of the compound Q68 to the alkali to the isoamyl nitrite is 1 (2-5): 2-5); preferably 1:5:5.

In the step (c), the temperature of the reaction is 25-50 ℃; preferably 25 ℃.

In the step (c), the reaction time is 3-8 hours; preferably 5h.

(D) Condensation reaction

Compound Q69 is dissolved in an organic solvent, hydroxylamine hydrochloride and alkali are added, and the compound Q70 is obtained through reaction.

In the step (d), the organic solvent is one or more of methanol, ethanol, pyridine and the like; preferably, pyridine.

In the step (d), the alkali is one or more of pyridine, triethylamine, na ₂CO₃、K₂CO₃, sodium acetate and the like; preferably, pyridine.

In the step (d), the molar ratio of the compound Q69 to hydroxylamine hydrochloride is 1 (2-5); preferably 1:3.

In the step (d), the temperature of the condensation reaction is 80-120 ℃; preferably at 100 ℃.

In the step (d), the time of the condensation reaction is 2-6 h; preferably 3h.

(E) Dehydration cyclization reaction

The compound Q70 is dissolved in an organic solvent, and alkali is added for reaction to obtain the compound Q71.

In the step (e), the organic solvent is selected from one or more of ethylene glycol, dioxane, mixed solvents of ethylene glycol and dioxane; preferably, the solvent is a mixed solvent of ethylene glycol and dioxane.

In the step (e), the optimal ratio of the ethylene glycol to the dioxane mixed solvent is 2:1.

In the step (e), the base of the dehydration cyclization reaction is selected from one or more of potassium hydroxide, sodium hydroxide, potassium acetate, sodium acetate and the like; preferably potassium hydroxide.

In the step (e), the temperature of the reaction is 100-220 ℃; preferably 130 ℃.

In the step (e), the reaction time is 3-8 h; preferably 5h.

(F) Acylation reaction

The synthesis of compound Q72 is similar to the acylation reaction of Q37 in scheme (4).

(G) Condensation reaction

And (3) dissolving the compound Q68 in an organic solvent, adding a condensing agent and diethyl oxalate, and reacting to obtain a compound Q73.

In the step (g), the organic solvent is selected from one or more of anhydrous tetrahydrofuran, dichloromethane, diethyl ether, carbon tetrachloride, benzene, toluene and the like; preferably anhydrous tetrahydrofuran.

In the step (g), the molar ratio of the compound Q68 to diethyl oxalate is 1 (1-5); preferably 1:2.

In the step (g), the condensing agent is selected from one or more of sodium hydride, sodium methoxide, sodium ethoxide and the like; preferably sodium hydride.

In the step (g), the molar ratio of the Q68 to the condensing agent is 1 (5-10); preferably 1:10.

In the step (g), the temperature of the condensation reaction is 25-45 ℃; preferably 25 ℃.

In the step (g), the reaction time of the condensation reaction is 20-60 min; preferably 20min.

(H) Addition cyclization reaction

The synthesis of compound Q74 is analogous to the cyclization reaction method of Q25 in scheme (4).

(I) Addition cyclization reaction

Dissolving the compound Q73 in an organic solvent, adding hydroxylamine hydrochloride, and reacting to obtain the compound Q75.

In the step (i), the organic solvent is selected from one or more of ethanol, ethanol-water mixed solvent, acetic acid and the like; preferably, it is a mixed solvent of ethanol and water.

In the step (i), the molar ratio of the compound Q73 to hydroxylamine hydrochloride is 1 (1-3); preferably 1:3.

In the step (i), the temperature of the reaction is 80-140 ℃; preferably at 100 ℃.

In the step (i), the reaction time is 3-10 h; preferably 3h.

The invention also provides a preparation method of the cholic acid derivative, which takes the compound Q24 as a starting material, and the compound Q79 is obtained through cyclization, oxidation, reduction and format reaction, wherein the preparation method is shown in a route (11):

(a) Addition cyclization reaction

The synthesis of compound Q76 is similar to the cyclization reaction method of Q75 in scheme (10).

(B) Oxidation reaction

The synthesis of compound Q77 is similar to the oxidation reaction method of Q47 in scheme (6).

(C) Reduction reaction

The synthesis of compound Q78 is similar to the reduction reaction method of Q48 in scheme (6).

(D) Format reaction

The synthesis of compound Q79 is similar to the format reaction method of Q56 in scheme (4).

The invention also provides a preparation method of the cholic acid derivative, which takes the compound Q68 as a starting material, and the compound Q81 is obtained through condensation and cyclization, wherein the preparation method is shown in a route (12):

(a) Condensation reaction

The synthesis of compound Q80 is similar to the condensation reaction method of Q24 in scheme (4).

(B) Cyclization reaction

And (3) dissolving the compound Q80 in an organic solvent, adding o-fluorine phenylhydrazine hydrochloride, and reacting to obtain the compound Q81.

In the step (b), the organic solvent is selected from one or more of ethanol, ethanol and water mixed solvent, acetic acid and the like; preferably, it is a mixed solvent of ethanol and water.

In the step (b), the molar ratio of the Q80 to the o-fluorine phenylhydrazine hydrochloride is 1 (1-3); preferably 1:3.

In the step (b), the temperature of the cyclization reaction is 80-140 ℃; preferably at 100 ℃.

In the step (b), the cyclization reaction time is 3-10 hours; preferably 3h.

The invention also provides a preparation method of the cholic acid derivative, which comprises the steps of taking the compound Q6 as a starting material, obtaining the compound Q82 through reduction, and then taking the Q82 as the starting material, and obtaining the cholic acid derivative Q87 through condensation, cyclization, hydrolysis and acylation; the preparation method is shown in a route (13):

(a) Reduction reaction

And dissolving the compound Q6 in an organic solvent, adding a reducing reagent, and carrying out a reduction reaction under pressure to obtain a compound Q82.

In the step (a), the organic solvent is selected from one or more of methanol, ethanol, tetrahydrofuran, dichloromethane, ethyl acetate and the like; preferably, ethyl acetate is used.

In the step (a), the reducing agent is selected from one or more of palladium carbon, raney nickel and the like; preferably palladium carbon

In the step (a), the mass ratio of the compound Q6 to the reducing agent is 1: (0.2-1); preferably, it is 1:0.5.

In step (a), the reaction pressure was 4MPa.

In the step (a), the temperature of the reaction is 10-60 ℃; preferably 25 ℃.

In the step (a), the reaction time is 24-72 h; preferably 24h.

(B) Condensation reaction

The synthesis of compound Q83 is similar to the condensation reaction method of Q24 in scheme (4).

(C) Addition cyclization reaction

The synthesis of compound Q84 is analogous to the cyclization reaction method of Q25 in scheme (4).

(D) Hydrolysis reaction

The synthesis of compound Q85 is similar to the hydrolysis reaction method of Q26 in scheme (4).

(E) Acetylation reaction

The synthesis of compound Q86 is similar to the acetylation reaction of Q27 in scheme (4).

(F) Acylation reaction

The synthesis of compound Q87 is similar to the acylation reaction of Q37 in scheme (4).

The invention also provides a preparation method of the cholic acid derivative, which takes the compound Q9, Q13, Q19 or Q23 as a starting material, and the compounds Q94-Q111 are obtained through condensation, cyclization, hydrolysis and acylation, and the preparation method is shown in a route (14):

(a) Aldol condensation reaction

The synthesis of compounds Q107, Q102, Q88, Q97 is similar to the condensation reaction method of Q24 in scheme (4).

(B) Addition cyclization reaction

The synthesis of compounds Q108, Q103, Q89, Q98 is similar to the cyclization reaction method of Q25 in scheme (4).

(C) Hydrolysis reaction

The synthesis of compounds Q109, Q104, Q90, Q99 is similar to the hydrolysis reaction method of Q26 in scheme (4).

(D) Acylation reaction

The synthesis of compounds Q110, Q105 is similar to the acylation reaction of Q54 in scheme (7).

The synthesis of compounds Q93, Q100 is similar to the acetylation reaction of Q27 in scheme (4).

(E) Acylation reaction

The synthesis of compounds Q111, Q106, Q96, Q101 is similar to the acylation reaction method of Q37 in scheme (4).

The synthesis of compound Q94 is similar to the acylation reaction of Q36 in scheme (4).

(F) Hydrolysis followed by acylation

The synthesis of compound Q95 is analogous to the reaction procedure of Q43 in scheme (5).

(G) Acylation reaction

The synthesis of compound Q91 is similar to the acylation reaction of Q37 in scheme (4).

(H) Acylation reaction

The synthesis of compound Q92 is similar to the acylation reaction of Q54 in scheme (7).

In the preparation method, the reaction is tracked and measured by a thin plate chromatography, and the post-treatment method adopted after the reaction is finished comprises the steps of concentration, extraction, column chromatography separation and the like, and the final product is verified by nuclear magnetic resonance spectrum and high-resolution mass spectrum.

The invention also provides application of the cholic acid derivative in preparing a medicament for inhibiting cell cholesterol synthesis.

The invention also provides application of the cholic acid derivative in preparing a medicament for preventing and/or treating hypercholesterolemia, hypertriglyceridemia, atherosclerosis and/or nonalcoholic steatohepatitis.

The medicament may also be used in combination with statin drugs.

The statin drugs comprise lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin and rosuvastatin.

The invention has the beneficial effects that: the cholic acid derivative disclosed by the invention not only can effectively promote the degradation of the hydroxymethyl glutaryl coenzyme A reductase (3-hydroxy-3-methyl-glutaryl-coenzymeAreductase, HMGCR), but also can reduce the increase of HMGCR protein caused by statin drugs, thereby reducing the endogenous cholesterol level, providing a beneficial reference for the research and development of new drugs for reducing cholesterol, resisting atherosclerosis and resisting nonalcoholic steatohepatitis, and having good application prospects.

Drawings

FIG. 1 shows the accumulation of intracellular cholesterol in the first-generation compounds.

FIG. 2 is a western plot of compound Q37 and compound Q54 and EC50 values of Q37 and Q54; wherein, figure a is a western plot of compound Q37, figure b is an EC50 of compound Q37, figure c is a western plot of compound Q54, and figure b is an EC50 of compound Q54.

FIG. 3 shows the accumulation of intracellular cholesterol (no accumulation at all) in the second-generation partial compounds.

Detailed Description

The present invention will be described in further detail with reference to the following specific examples and drawings. The procedures, conditions, experimental methods, etc. for carrying out the present invention are common knowledge and common knowledge in the art, except for the following specific references, and the present invention is not particularly limited.

The structures of the compounds in the following examples were determined by nuclear magnetic resonance; reagents are mainly provided by Shanghai national pharmaceutical chemical reagent company; the product is purified mainly by column chromatography, silica gel (200-300 mesh) produced by Qingdao ocean chemical plant.

EXAMPLE 1 preparation of Compounds Q2-Q4

Compound Q1 (10 g,0.03 mol) was placed in a 250mL single-neck flask, 150mL of anhydrous DMF was added and stirred to dissolve, imidazole (16.34 g,0.24 mol), TBSCl (18.09 g,0.12 mol) and N ₂ were added at 0 ℃ to displace three times, after stirring evenly and then to room temperature, stirring was carried out for 5 hours, after TLC detection of complete reaction of the starting materials, the reaction was quenched by adding 50mL of saturated ammonium chloride solution, ethyl acetate (100 mL) was added to extract the separated liquid, the aqueous phase was extracted again with ethyl acetate (30 ml×3), and the organic phases were combined. The organic phase was washed with saturated ammonium chloride solution (50 mL. Times.2), water (50 mL. Times.3) and saturated sodium chloride solution (50 mL. Times.2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure to give Compound Q2 (white solid 13.45g, yield 99%) which was used directly in the next step.

Compound Q2 (13.45 g,0.03 mol) was placed in a three-necked flask, 200mL of anhydrous tertiary butanol was added and stirred until it was dissolved, N ₂ was replaced three times, potassium tertiary butoxide (13.47 g,0.12 mol) was added in portions at 0 ℃ until it was clear, the visible system turned orange-yellow, methyl iodide (14 mL,0.24 mol) was slowly added dropwise, the visible system gradually turned pale yellow to white, the reaction was exothermic, the transfer to room temperature and stirring was carried out for 4 hours, TLC detected complete reaction of the starting material, saturated sodium sulfite solution (50 mL) was added to quench the reaction, water (50 mL) and ethyl acetate (100 mL) were added to extract fractions, the aqueous phase was extracted again with ethyl acetate (30 ml×3), and the organic phases were combined. The organic phase was washed with water (50 mL. Times.3) and saturated sodium chloride solution (50 mL. Times.2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure to give compound Q3 (white solid) which was used directly in the next step.

Compound Q3 was directly dissolved in diethyl ether hydrochloride solution, stirred at room temperature for 2h, tlc detected complete reaction of the starting materials, 50mL of water was added, 100mL of ethyl acetate was added for extraction of the separated liquid, the aqueous phase was extracted with ethyl acetate (30 ml×3), and the organic phases were combined. The organic phase was washed with water (50 ml×3), saturated sodium chloride solution (50 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure followed by silica gel column chromatography (PE: ea=20:1) to give compound Q4 (6.88 g white solid, two-step yield 64％).¹³C NMR(101MHz,CDCl3)δ217.12,149.52,119.47,67.90,56.44,52.38,48.87,48.67,42.46,39.58,38.73,37.05,33.71,32.07,31.70,31.22,30.22,27.76,27.23,24.28,21.27,19.33,16.78,11.99.

EXAMPLE 2 preparation of Compound Q5

Compound Q4 (6.88 g,0.019 mol), PDC (10.84 g,0.029 mol) and silica gel (10.84 g,0.029 mol) were placed in a 250mL single-neck flask, 100mL of anhydrous methylene chloride was added, stirring was performed at room temperature for 8 hours, TLC detection of the starting material was complete, the silica gel was removed by suction filtration, the filter cake was washed with methylene chloride (20 mL. Times.3), saturated sodium sulfite was added to quench the system for ten minutes until the system became pale green, the extract was separated, the aqueous phase was extracted with methylene chloride (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (30 ml×3), saturated sodium chloride solution (30 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=15:1) to give compound Q5 (white solid 6.83g, yield 99％).¹³C NMR(101MHz,CDCl3)δ216.74,205.03,149.81,119.83,56.02,50.96,49.49,48.88,48.66,42.99,39.43,37.05,33.68,32.08,31.65,31.20,30.22,27.23,27.11,24.55,21.22,19.33,13.44,12.28.

EXAMPLE 3 preparation of Compound Q6

NaH (3.42 g,0.142 mol) was added to a 250mL three-necked flask in an ice bath, N ₂ was substituted three times, 50mL anhydrous THF was added, stirring was performed to dissolve, triethyl phosphorylacetate (18.85 mL,0.095 mol) was slowly dropped, and the system was observed to generate bubbles after about 10 minutes. The system was brought to room temperature and stirred for 10 minutes. Compound Q5 (6.83 g,0.019 mol) was dissolved in anhydrous THF (50 mL), the above system was added dropwise, stirred at room temperature for 1 hour, TLC was used to detect complete reaction of the starting materials, 50mL of saturated ammonium chloride solution was added to quench for ten minutes, 60mL of ethyl acetate was added to extract the separated liquid, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with saturated ammonium chloride (30 ml×2), water (30 ml×3), saturated sodium chloride solution (30 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=15:1) to give compound Q6 (7.94 g of a white solid, yield 98％).¹³C NMR(101MHz,CDCl3)δ216.72,167.06,154.60,149.78,119.83,118.99,60.11,56.56,54.88,48.86,48.65,42.70,39.72,39.57,37.05,33.69,32.07,31.64,31.19,30.22,28.19,27.22,24.18,21.25,19.33,19.22,14.29,12.15.

EXAMPLE 4 preparation of Compound Q7

Compound Q6 (7.94 g,0.019 mol) and magnesium metal turnings (4.56 g,0.19 mol) were placed in a 250mL single-necked flask, 100mL of anhydrous methanol was added, and the flask was stirred at room temperature for 5 hours, and the system was seen to be cloudy, generate bubbles, and generate heat in the flask body. TLC was used to check the completion of the reaction, water (30 mL), ethyl acetate (50 mL), 2M dilute hydrochloric acid was added to the solid solution, the solution was separated by extraction, the aqueous phase was extracted with ethyl acetate (30 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (30 mL. Times.3) and saturated sodium chloride solution (30 mL. Times.2), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and dried to give a white solid. The white solid was placed in a 250mL single-neck flask, toluene was added to the solution, IBX (5.32 g,0.019 mol) was added, DMSO was added to the basic solution, stirring was performed at room temperature for 4 hours, TLC was used to detect complete reaction of the starting material, 50mL of water was added, 50mL of ethyl acetate was used, a large amount of insoluble solid was found to precipitate, suction filtration was performed, the filter cake was washed with ethyl acetate (20 mL. Times.3), the solution was separated, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (50 ml×5), saturated sodium chloride solution (30 ml×3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=15:1) to give compound Q7 (white solid 7.08g, yield 90％).¹³C NMR(101MHz,CDCl3)δ216.73,174.36,149.51,119.83,56.72,55.74,51.49,48.90,48.60,42.40,39.70,37.04,35.35,33.70,32.06,31.68,31.17,31.03,30.97,30.21,28.13,27.22,24.14,21.26,19.32,18.29,11.91.

EXAMPLE 5 preparation of Compound Q8

Potassium permanganate (2 g,12.66 mmol) was dissolved in a 100mL beaker with 30mL of water added and stirred. Tetrabutylammonium bromide (4.3 g,13.34 mmol) was put into a 50mL beaker, water (8 mL) was added, the above system was added dropwise to give a purple suspension, the suspension was stirred at room temperature for 40 minutes and then filtered with suction, the filter cake was washed with water (20 mL. Times.3), and the solid tetrabutylammonium permanganate was obtained after air drying at room temperature for use.

In a 100mL single-neck flask, 40mL of pyridine was added to compound Q5 (2 g,5.61 mmol) and tetrabutylammonium permanganate (4.05 g,11.22 mmol) prepared as described above, stirred for thirty minutes and then brought to room temperature, stirring was continued for 1 hour, TLC was used to detect complete reaction of the starting materials, water (20 mL), saturated sodium bisulfite solution (30 mL) was added, pH was adjusted to approximately 5 with 2M dilute hydrochloric acid, 50mL ethyl acetate was added to extract the separated liquid, the aqueous phase was extracted again with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (30 ml×3), saturated sodium chloride solution (30 ml×3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=10:1) to give compound Q8 (white solid 1.98g, yield 95％).¹³C NMR(126MHz,CDCl3)δ216.90,182.52,149.78,119.81,56.38,52.43,48.85,48.69,42.55,42.48,39.56,37.06,33.70,32.07,31.64,31.22,30.23,27.34,27.23,24.27,21.23,19.34,17.07,12.09.

EXAMPLE 6 preparation of Compound Q9

Compound Q8 (1.98 g,5.33 mmol) was added to a 100mL single-neck flask, 50mL of methanol was added, stirring was uniform, concentrated sulfuric acid (0.2 mL,3.73 mmol) was slowly added dropwise, heating was performed at 70℃under reflux, stirring was performed for 2 hours, TLC was used to detect complete reaction of the starting material, water (20 mL) was added, ethyl acetate (50 mL) was added to extract fractions, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (30 ml×3), saturated sodium chloride solution (30 ml×3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=15:1) to give compound Q9 (white solid 2.06g, yield 99％).¹³C NMR(126MHz,CDCl₃)δ216.73,177.31,149.79,119.81,56.35,52.84,51.35,48.86,48.67,42.46,42.44,39.52,37.05,33.70,32.07,31.64,31.21,30.22,27.23,27.20,24.23,21.23,19.33,17.13,12.08.

EXAMPLE 7 preparation of Compounds Q10 to Q11

Iodine simple substance (3.47 g,13.7 mmol) was dissolved in a 100mL three-necked flask, anhydrous toluene (30 mL) was added, triphenylphosphine (3.74 g,14.3 mmol) and imidazole (1.86 g,27.4 mmol) were added, nitrogen protection was provided, the visible system was visibly discolored, and the mixture was stirred at room temperature for 1 hour to obtain a brown-yellow system. To the above system was added compound Q4 (700 mg,1.95 mmol), stirred at room temperature for 2 hours, and TLC was used to check that the starting material was complete, quenched by adding saturated sodium sulfite solution (20 mL) for ten minutes to almost colorless, extracted with ethyl acetate (30 mL) and separated, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (30 mL. Times.3) and saturated sodium chloride solution (30 mL. Times.3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure to give compound Q10 (white solid), which was taken directly to the next step.

Compound Q10 (all solids from the previous step) was placed in a 100mL single-neck flask, 30mL of ldmf was added, the solution was stirred, KF (1.81 g,31.2 mmol), trimethylsilyl cyanide (1.95 mL,15.6 mmol) were added, the reaction was complete by TLC at 50 ℃ for 4 hours, 30mL of water was added, 30mL of ethyl acetate was extracted for separation, the aqueous phase was extracted with ethyl acetate (20 ml×3), and the organic phases were combined. The organic phase was washed with water (30 ml×3), saturated sodium chloride solution (30 ml×3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure followed by silica gel column chromatography (PE: ea=15:1) to give compound Q11 (white solid 537mg, two-step yield 75％).¹³C NMR(126MHz,CDCl₃)δ216.71,149.82,119.76,118.94,56.53,54.69,48.72,48.66,42.49,39.35,37.04,33.69,33.53,32.05,31.61,31.19,30.22,28.11(2C),27.23,24.82,24.07,21.19,19.33,12.01.

EXAMPLE 8 preparation of Compound Q12

Compound Q11 (537 mg,1.46 mmol) and NaOH (284 mg,14.6 mmol) were placed in a 50mL single-necked flask, a mixed solution (20 mL) of water and ethanol (1:3) was added, the system was stirred to be substantially dissolved, the mixture was heated under reflux at 100℃for 72 hours, the reaction of the starting materials was detected by TLC, the pH was adjusted to about 5 by adding a 2M diluted hydrochloric acid solution, extraction and separation of the mixture was performed by adding ethyl acetate (20 mL), the aqueous phase was extracted with ethyl acetate (10 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=10:1) to give compound Q12 (white solid 507mg, yield 90％).¹³C NMR(126MHz,CDCl₃)δ216.99,179.43,149.79,119.85,56.76,55.75,48.84,48.69,42.51,41.28,39.59,37.06,33.71,33.61,32.08,31.67,31.20,30.21,28.27,27.25,24.12,21.24,19.53,19.34,11.94.

EXAMPLE 9 preparation of Compound Q13

Compound Q12 (507 mg,1.31 mmol) was placed in a 50mL single-neck flask, methanol (20 mL) was added, stirring was uniform, concentrated sulfuric acid (0.05 mL,0.92 mmol) was slowly added dropwise, heating was performed at 70℃under reflux, stirring was performed for 2 hours, TLC was used to detect complete reaction of the starting material, water (10 mL) was added, ethyl acetate (20 mL) was added to extract the separated liquid, the aqueous phase was extracted with ethyl acetate (10 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (10 ml×3), saturated sodium chloride solution (10 ml×3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=15:1) to give compound Q13 (white solid 524mg, yield 99％).¹³C NMR(126MHz,CDCl₃)δ216.74,173.97,149.81,119.85,56.75,55.89,51.36,48.85,48.66,42.51,41.42,39.60,37.05,33.77,33.71,32.09,31.68,31.20,30.20,28.23,27.25,24.13,21.24,19.55,19.33,11.95.

EXAMPLE 10 preparation of Compound Q14

In a 100mL single-necked flask, compound Q7 (1 g,2.42 mmol) and ethylene glycol (10 mL) were added anhydrous THF (30 mL), p-toluenesulfonic acid (96 mg, 0.284 mmol), triethyl orthoformate (2 mL,12.1 mmol), and N ₂ were added, followed by stirring at room temperature for 3 hours, and the reaction was complete by TLC, water (30 mL) was added, ethyl acetate (50 mL) was added, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=20:1) to give compound Q14 (white solid 1.11g, yield 99％).¹³C NMR(126MHz,CDCl₃)δ174.86,149.64,119.89,113.22,65.22,64.87,57.17,55.57,51.49,50.54,44.82,42.27,39.70,36.26,35.35,35.26,32.30,31.04,31.00,30.88,29.20,28.15,26.83,24.15,22.39,21.70,20.54,18.29,11.86.

EXAMPLE 11 preparation of Compound Q15

Compound Q14 (1.11 g,2.42 mmol) was added to anhydrous THF (40 mL) and dissolved under stirring, lithium aluminum hydride (919 mg,24.2 mmol) was added in portions at 0deg.C, nitrogen blanket, stirring was continued for 2 hours, TLC checked for complete reaction of the starting material, 0.92mL of water, 2M sodium hydroxide solution (1.85 mL), quenched with water (0.92 mL), extracted with water (30 mL) and ethyl acetate (30 mL), the aqueous phase was extracted again with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=20:1) to give compound Q15 (white solid 1.04g, yield 99％).¹H NMR(500MHz,CDCl₃)δ5.54–5.49(m,1H),4.03–3.86(m,4H),3.67–3.54(m,2H),2.09–2.04(m,1H),2.01–1.96(m,2H),1.86–1.80(m,1H),1.74–1.37(m,16H),1.27(dd,J＝9.0,6.0Hz,1H),1.23(s,3H),1.12(s,3H),1.07(dd,J＝5.3,3.0Hz,1H),1.04(s,3H),0.96(dd,J＝5.1,3.3Hz,1H),0.93(d,J＝6.5Hz,3H),0.67(s,3H).

EXAMPLE 12 preparation of Compound Q16

Iodine simple substance (4.31 g,16.94 mmol) was dissolved in a 100mL three-necked flask, anhydrous toluene (30 mL) was added, triphenylphosphine (4.63 g,17.67 mmol) and imidazole (2.31 g,33.88 mmol) were added, nitrogen protection was provided, the visible system was visibly discolored, and the mixture was stirred at room temperature for 1 hour to obtain a brown-yellow system. To the above system was added compound Q15 (1.04 g,2.42 mmol), stirred at room temperature for 2 hours, and TLC checked for complete reaction of the starting materials, quenched for ten minutes with saturated sodium sulfite solution (30 mL) to almost colorless, extracted with ethyl acetate (30 mL), separated by extraction of the aqueous phase with ethyl acetate (20 mL. Times.3), and the organic phases combined. The organic phase was washed with water (30 ml×3), saturated sodium chloride solution (30 ml×3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=30:1) to give compound Q16 (white solid 1.18g, yield 90％).¹³CNMR(126MHz,CDCl₃)δ149.68,119.89,113.20,65.24,64.89,57.19,55.68,50.56,44.84,42.28,39.71,36.89,36.28,35.28,35.08,32.32,30.89,30.35,29.22,28.28,26.85,24.18,22.41,21.72,20.56,18.75,11.88,7.89.

EXAMPLE 13 preparation of Compound Q17

Compound Q16 (1.18 g,2.18 mmol) was placed in a 100mL single-neck flask, DMF (30 mL) was added, the solution was stirred, KF (2.02 g,34.85 mmol) and trimethylsilylcyanide (2.18 mL,17.42 mmol) were added, the reaction was complete as detected by TLC at 50℃for 4 hours, water (30 mL) and ethyl acetate (30 mL) were added to extract fractions, the aqueous phase was extracted again with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (30 ml×3), saturated sodium chloride solution (30 ml×3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=15:1) to give compound Q17 (white solid 813mg, yield 85％).¹³C NMR(126MHz,CDCl₃)δ149.68,119.87,113.20,65.24,64.89,57.17,55.58,50.54,44.84,42.29,39.72,36.27,35.27,35.23,35.05,32.29,30.88,29.22,28.26,26.84,24.14,22.40,22.19,21.71,20.55,18.52,17.55(2C),11.86.

EXAMPLE 14 preparation of Compound Q18

Compound Q17 (813 mg,1.85 mmol) and NaOH (741mg, 18.5 mmol) were placed in a 50mL single-necked flask, a mixed solution (30 mL) of water and ethanol (1:3) was added, the mixture was stirred to substantially dissolve the compound, the mixture was heated to reflux at 100℃for 72 hours, TLC was used to detect completion of the reaction of the starting material, 2M diluted hydrochloric acid solution was added to adjust pH to about 5, 30mL of ethyl acetate was added to extract the separated liquid, and the aqueous phase was extracted with ethyl acetate (15 mL. Times.3) and the organic phases were combined. The organic phase was washed with water (20 mL. Times.3) and saturated sodium chloride solution (20 mL. Times.3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure to give Compound Q18 (690 mg of white solid, yield) 90％).¹H NMR(500MHz,CDCl₃)δ5.54(s,1H),2.56–2.42(m,2H),2.30(dt,J＝23.4,7.9Hz,2H),2.09(dt,J＝17.6,4.8Hz,1H),2.00(t,J＝11.0Hz,2H),1.86–1.79(m,1H),1.70–1.35(m,10H),1.26(d,J＝8.2Hz,1H),1.22(s,6H),1.17–1.04(m,4H),1.04–0.98(m,2H),0.93(d,J＝6.2Hz,3H),0.84(s,3H),0.67(s,3H).

EXAMPLE 15 preparation of Compound Q19

Compound Q18 (460 mg,1.67 mmol) was placed in a single-neck flask, methanol (20 mL) was added, stirring was uniform, concentrated sulfuric acid (0.06 mL,1.17 mmol) was slowly added dropwise, heating was performed at 70℃under reflux, stirring was performed for 2 hours, TLC was used to detect complete reaction of the starting material, water (10 mL) and ethyl acetate (20 mL) were added to extract fractions, the aqueous phase was extracted with ethyl acetate (15 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (15 ml×3), saturated sodium chloride solution (15 ml×3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=15:1) to give compound Q19 (715 mg as a white solid, yield 99％).¹³C NMR(126MHz,CDCl₃)δ216.84,174.32,149.79,119.91,56.76,55.77,51.43,48.90,48.66,42.38,39.72,37.06,35.48,35.39,34.51,33.72,32.09,31.72,31.20,30.21,28.19,27.24,24.16,21.50,21.28,19.34,18.59,11.90.

EXAMPLE 15 preparation of Compound Q20

Compound Q15 (1 g,2.33 mmol) was placed in a 100mL single-neck flask, toluene was added to the solution, IBX (977 mg,2.49 mmol) was added, then DMSO was added to the basic solution, stirring was performed at room temperature for 8 hours, TLC was used to detect complete reaction of the starting materials, water (30 mL) and ethyl acetate (30 mL) were added, a large amount of insoluble solids were found to precipitate, suction filtration was performed, the filter cake was washed with ethyl acetate (20 mL. Times.3), the solution was separated, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (50 ml×5), saturated sodium chloride solution (30 ml×3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=20:1) to give compound Q20 (white solid 998mg, yield 99％).¹³C NMR(126MHz,CDCl₃)δ203.23,149.68,119.86,113.20,65.24,64.88,57.17,55.62,50.53,44.84,42.30,40.90,39.70,36.27,35.33,35.27,32.30,30.89,29.22,28.23,27.97,26.84,24.16,22.40,21.70,20.55,18.41,11.89.

EXAMPLE 16 preparation of Compound Q21

NaH (319 mg,17.48 mmol) was placed in a 100mL three-necked flask under ice bath, N ₂ was substituted three times, 20mL anhydrous THF was added, stirring was performed to dissolve, triethyl phosphorylacetate (2.31 mL,11.65 mmol) was slowly dropped, and the system was observed to generate bubbles after about 10 minutes. The system was brought to room temperature and stirred for 10 minutes. Compound Q20 (998 mg,2.33 mmol) was dissolved in anhydrous THF (20 mL), the above system was added dropwise, stirred at room temperature for 1 hour, TLC was checked for complete reaction of the starting materials, saturated ammonium chloride solution (30 mL) was added and quenched for ten minutes, ethyl acetate (30 mL) was added to extract the separated liquid, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with saturated ammonium chloride (20 ml×2), water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=15:1) to give compound Q21 (1.14 g of a white solid, yield 98％).¹³C NMR(126MHz,CDCl₃)δ166.78,149.95,149.66,120.96,119.85,113.17,65.22,64.86,60.08,57.17,55.73,50.53,44.82,42.29,39.72,36.26,35.46,35.25,34.28,32.30,30.88,29.21,28.97,28.23,26.83,24.15,22.39,21.70,20.55,18.45,14.28,11.88.

EXAMPLE 17 preparation of Compound Q22

Compound Q21 (1.14 g,2.33 mmol) was put in a one-necked flask, and the aqueous methanol mixture was stirred until it was substantially dissolved. 2M dilute aqueous hydrochloric acid (6 mL) was added to the system, stirred at room temperature for 2 hours, TLC was used to detect complete reaction of the starting materials, ethyl acetate (30 mL) was added to extract the separated solution, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=15:1) to give compound Q22 (1.06 g of white solid, yield 99％).¹³C NMR(126MHz,CDCl₃)δ216.86,166.81,149.91,149.78,121.01,119.91,60.13,56.75,55.91,48.89,48.67,42.44,39.74,37.06,35.47,34.28,33.72,32.09,31.70,31.20,30.21,29.00,28.22,27.25,24.15,21.28,19.34,18.45,14.29,11.95.

EXAMPLE 18 preparation of Compound Q23

Compound Q22 (1.06 g,2.33 mmol) and magnesium turnings (560 mg,23.3 mmol) were placed in a single-necked flask, and anhydrous methanol (40 mL) was added thereto, followed by stirring at room temperature for 5 hours, whereupon the visible system became cloudy, and bubbles were generated, and the body of the flask became hot. TLC was used to check the completion of the reaction, water (20 mL), ethyl acetate (30 mL), 2M dilute hydrochloric acid was added to the solid solution, the solution was separated by extraction, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (20 mL. Times.3) and saturated sodium chloride solution (20 mL. Times.2), dried over anhydrous Na ₂SO₄, concentrated under reduced pressure, and dried to give a white solid. The white solid was placed in a 100mL single-neck flask, toluene was added to the solution, IBX (653 mg,2.33 mmol) was added, DMSO was added to the basic solution, stirring was performed at room temperature for 4 hours, TLC was used to detect complete reaction of the starting materials, water (30 mL) and ethyl acetate (30 mL) were added, a large amount of insoluble solid was found to precipitate, suction filtration was performed, the filter cake was washed with ethyl acetate (20 mL. Times.3), the solution was separated, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (30 ml×5), saturated sodium chloride solution (30 ml×3), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=15:1) to give compound Q23 (white solid 928mg, yield 90％).¹³C NMR(126MHz,CDCl₃)δ216.83,174.30,149.78,119.92,56.77,56.03,51.43,48.91,48.66,42.37,39.74,37.06,35.57,35.51,34.16,33.72,32.09,31.73,31.20,30.20,28.25,27.25,25.65,25.39,24.17,21.28,19.33,18.62,11.92.

EXAMPLE 19 preparation of Compounds Q24-Q25

Compound Q7 (1.00 g,2.41 mmol) was placed in a 100mL single-necked flask, 30mL ethyl formate was added to the solution, naH (578 mg,24.1 mmol) was added, the mixture was stirred at room temperature for 20 minutes under nitrogen protection, the visible system turned orange-yellow, TLC was used to detect complete reaction of the starting material, 30mL water, 30mL ethyl acetate, the aqueous phase was extracted once more with 20mL ethyl acetate, and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure to give a pale yellow oil, Q24, which was unstable and was immediately followed.

Compound Q24 (previous step oil) was added to a 100mL single-neck flask, 30mL of glacial acetic acid solution, potassium acetate (473 mg,4.82 mmol) and 85% hydrazine hydrate (0.42 mL,8.51 mmol) were added, heated to reflux for 5 hours at 130 ℃ and TLC detected complete reaction of the starting materials, cooled to room temperature, poured into a beaker, stirred for 10 minutes with 2M sodium hydroxide solution (100 mL), stirred for 10 minutes with saturated sodium bicarbonate solution, ethyl acetate (30 mL) was added to extract fractions, the aqueous phase was extracted again with ethyl acetate (20 ml×3) and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=5:1) to give compound Q25 (white solid 1g, yield 91.8％).¹HNMR(500MHz,DMSO)δ12.32(s,1H),7.20(s,1H),5.71(t,J＝13.9Hz,1H),4.11–3.96(m,2H),2.63(t,J＝24.9Hz,1H),2.29(td,J＝9.5,4.6Hz,1H),2.21–2.14(m,1H),2.10–1.99(m,3H),1.84–1.79(m,1H),1.72–1.63(m,2H),1.64–1.55(m,2H),1.52–1.43(m,2H),1.40(d,J＝7.5Hz,3H),1.34(t,J＝9.0Hz,1H),1.27(s,3H),1.24–1.20(m,3H),1.19–1.14(m,4H),1.11–1.01(m,3H),0.89(t,J＝8.1Hz,3H),0.79(s,3H),0.66(s,3H).

EXAMPLE 20 preparation of Compound Q26

In a 100mL single-necked flask, 40mL of a mixed solution of aqueous ethanol and lithium hydroxide (1 g,2.21 mmol) was added, the reaction was completed by TLC, 2M diluted hydrochloric acid aqueous solution was added to adjust pH to about 5, ethyl acetate (30 mL) was added, the extract was separated, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phase was combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure followed by silica gel column chromatography (DCM/meoh=15:1) to give compound Q26 (white solid 1.03g, yield 99％).¹HNMR(400MHz,DMSO)δ12.05(s,1H),7.19(s,1H),5.68(d,J＝25.5Hz,1H),2.64(d,J＝14.4Hz,1H),2.21(s,1H),2.02(dd,J＝30.1,15.3Hz,4H),1.88–1.50(m,6H),1.45(s,1H),1.39(s,3H),1.33(s,1H),1.25(s,3H),1.22–0.98(m,7H),0.87(d,J＝5.2Hz,3H),0.77(s,3H),0.66(d,J＝14.9Hz,3H).mp:260-261℃.HRMS(ESI)for C₂₇H₄₁N₂O₂[M+H]⁺:calcd 425.3163,found425.3184.

EXAMPLE 21 preparation of Compound Q27

Compound Q26 (1.03 g,2.21 mmol), DMAP (73.37 mg,0.44 mmol) were placed in a 100mL single-necked flask, THF (40 mL), acetic anhydride (0.63 mL,6.63 mmol) were added, the mixture was heated and stirred at 60℃for 2 hours, the visible system was gradually cleared, the reaction of the starting materials was checked by TLC and was returned to room temperature, water (30 mL) and ethyl acetate (30 mL) were added to extract the separated liquid, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=2:1) to give compound Q27 (white solid 927mg, yield 90％).¹H NMR(500MHz,CDCl₃)δ7.88(d,J＝1.2Hz,1H),5.75(dd,J＝5.2,2.3Hz,1H),2.77(d,J＝14.9Hz,1H),2.65(s,3H),2.44–2.35(m,1H),2.33–

2.24(m,1H),2.18–2.09(m,2H),2.08–2.00(m,1H),1.93–1.77(m,2H),1.72–1.54(m,4H),1.52(s,3H),1.50–1.39(m,2H),1.35(s,3H),1.31–1.00(m,7H),0.94(t,J＝10.0Hz,3H),0.79(s,3H),0.70(s,3H).¹³C NMR(126MHz,CDCl₃)δ180.26,169.67,162.93,149.25,124.04,121.19,119.29,56.85,55.73,49.12,42.31,39.71,38.57,36.35,35.32,33.78,32.24,31.94,31.80,31.59,31.02,30.77,28.14,24.19,21.63,21.14,21.01,18.30,11.92.mp:104-105℃.HRMS(ESI)for C₂₉H₄₂N₂O₃[M+Na]⁺:calcd 489.3088,found 489.3099.

EXAMPLE 22 preparation of Compounds Q28-Q42

Compound Q27 (145 mg,0.31 mmol), EDCI (120.8 mg,0.63 mmol), HOBt (85.2 mg,0.63 mmol), DMAP (154.0 mg,1.26 mmol), and the corresponding amine (0.59 mmol) were placed in a 100mL single-neck flask, anhydrous DCM (10 mL) was added, stirring at room temperature for 12 hours, after completion of the reaction by TLC, 20mL of water was added, the aqueous phase was extracted with DCM (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (30 mL), saturated NaCl solution (30 mL), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography to give the corresponding compound.

Compound Q28, white solid, yield 95％.¹H NMR(500MHz,CDCl₃)δ7.87(d,J＝1.3Hz,1H),5.74(dd,J＝5.2,2.3Hz,1H),5.53(s,1H),2.80(d,J＝4.8Hz,3H),2.76(d,J＝14.9Hz,1H),2.64(s,3H),2.28–2.18(m,1H),2.17–2.10(m,2H),2.08–2.01(m,2H),1.95–1.84(m,1H),1.84–1.77(m,1H),1.71–1.54(m,4H),1.52(s,3H),1.50–1.39(m,2H),1.34(d,J＝6.9Hz,3H),1.32–1.00(m,7H),0.93(d,J＝6.5Hz,3H),0.78(s,3H),0.68(s,3H).¹³C NMR(126MHz,CDCl₃)δ

174.17,169.63,162.91,149.22,123.99,121.21,119.28,56.85,55.82,49.12,42.30,39.73,38.56,36.34,35.55,33.78,33.56,32.23,31.93,31.83,31.80,31.58,28.21,26.32,24.20,21.63,21.14,21.00,18.43,11.91.mp:178-180℃.HRMS(ESI)for C₃₀H₄₅N₃O₂[M+Na]⁺:calcd 502.3404,found 502.3409.

Compound Q29, white solid, yield 95％.¹H NMR(500MHz,CDCl₃)δ7.86(s,1H),5.74(dd,J＝5.1,2.1Hz,1H),5.51(s,1H),3.35–3.21(m,2H),2.75(d,J＝14.9Hz,1H),2.64(s,3H),2.25–

2.18(m,1H),2.17–2.09(m,2H),2.09–2.00(m,2H),1.91–1.85(m,1H),1.84–1.75(m,1H),1.72–1.54(m,4H),1.51(s,3H),1.49–1.38(m,2H),1.33(d,J＝7.2Hz,3H),1.30–1.15(m,4H),1.12(t,J＝7.3Hz,4H),1.08–0.99(m,2H),0.92(t,J＝9.3Hz,3H),0.79(d,J＝10.8Hz,3H),0.68(s,3H).¹³C NMR(126MHz,CDCl₃)δ173.37,169.62,162.91,149.22,123.99,121.21,119.28,56.85,55.83,49.12,42.30,39.73,38.56,36.33,35.53,34.31,33.78,33.69,32.23,31.93,31.83,31.80,31.58,28.22,24.19,21.63,21.14,21.00,18.45,14.94,11.91.mp:125-126℃.HRMS(ESI)for C₃₁H₄₇N₃O₂[M+Na]⁺:calcd 516.3560,found 516.3578.

Compound Q30, white solid, yield 95％.¹H NMR(400MHz,CDCl₃)δ7.86(s,1H),5.73(s,1H),5.60(s,1H),3.22–3.12(m,2H),2.72(d,J＝19.5Hz,1H),2.63(s,3H),2.26–2.17(m,1H),2.11(d,J＝15.4Hz,2H),2.02(d,J＝12.5Hz,2H),1.91–1.54(m,6H),1.51(s,3H),1.44–1.37(m,3H),1.33(s,3H),1.29–1.05(m,7H),0.93–0.85(m,7H),0.77(s,3H),0.67(s,3H).¹³C NMR(101MHz,CDCl₃)δ173.58,169.64,162.93,149.21,123.99,121.21,119.29,56.85,55.83,49.12,42.29,41.20,39.72,38.55,36.33,35.52,33.77,33.71,32.22,31.92,31.87,31.79,31.57,28.21,24.19,22.90,21.62,21.13,20.99,18.43,11.90,11.37.mp:125-126℃.HRMS(ESI)for C₃₂H₄₉N₃O₂[M+Na]⁺:calcd 530.3717,found 530.3707.

Compound Q31, white solid, yield 90％.¹H NMR(500MHz,CDCl₃)δ8.56(d,J＝2.2Hz,1H),8.32(d,J＝4.2Hz,1H),8.20(d,J＝8.2Hz,1H),8.03(s,1H),7.87(d,J＝1.0Hz,1H),7.26(t,J＝5.1Hz,1H),5.75(dd,J＝5.1,2.2Hz,1H),2.76(d,J＝14.9Hz,1H),2.65(s,3H),2.51–2.41(m,1H),2.35–2.24(m,1H),2.17–2.10(m,2H),2.03(dd,J＝9.2,3.3Hz,1H),1.94–1.86(m,2H),1.70–1.55(m,4H),1.52(s,3H),1.49–1.42(m,2H),1.34(s,3H),1.31–1.00(m,7H),0.96(d,J＝6.2Hz,3H),0.78(s,3H),0.67(s,3H).¹³C NMR(126MHz,CDCl₃)δ172.57,169.75,162.98,149.20,144.91,140.97,135.20,127.18,123.99,123.74,121.20,119.33,56.85,55.83,49.11,42.33,39.75,38.56,36.35,35.53,34.45,33.78,32.24,31.92,31.81,31.58,31.50,28.25,24.19,21.66,21.15,21.01,18.49,11.94.mp:174-175℃.HRMS(ESI)for C₃₄H₄₆N₄O₂[M+H]⁺:calcd543.3694,found 543.3697.

Compound Q32, white solid, yield 90％.¹H NMR(400MHz,CDCl₃)δ7.87(s,1H),6.00(s,1H),5.72(d,J＝21.5Hz,1H),3.71(s,4H),3.35(d,J＝5.2Hz,2H),2.76(d,J＝14.8Hz,1H),2.64(s,2H),2.51–2.42(m,5H),2.30–2.21(m,1H),2.16–1.56(m,11H),1.52(s,3H),1.44(d,J＝8.9Hz,1H),1.34(d,J＝6.6Hz,3H),1.29–1.02(m,9H),0.95(d,J＝6.1Hz,3H),0.78(s,3H),0.68(s,3H).¹³C NMR(101MHz,CDCl₃)δ173.62,169.66,162.93,149.22,124.02,121.21,119.29,66.91(2C),57.10,56.85,55.82,53.33(2C),49.10,42.30,39.72,38.56,36.34,35.52,33.78,33.56,32.22,31.93,31.80,31.77,31.58,29.70,28.23,24.20,21.65,21.13,21.01,18.43,11.93.mp:108-109℃.HRMS(ESI)for C₃₅H₅₄N₄O₃[M+H]⁺:calcd 579.4269,found 579.4215.

Compound Q33, white solid, yield 95％.¹H NMR(500MHz,CDCl₃)δ7.87(s,1H),7.79(d,J＝7.5Hz,1H),7.19(dd,J＝15.4,7.5Hz,2H),7.06(dd,J＝15.6,7.8Hz,1H),7.01(d,J＝18.7Hz,1H),5.76(dd,J＝5.1,2.2Hz,1H),2.77(d,J＝14.9Hz,1H),2.65(s,3H),2.50–2.40(m,1H),2.31(dd,J＝14.9,8.8Hz,1H),2.26(s,3H),2.17–2.10(m,2H),2.09–2.03(m,1H),1.93(s,2H),1.74–1.55(m,4H),1.53(s,3H),1.51–1.42(m,2H),1.36(s,3H),1.34–1.04(m,6H),1.01(d,J＝7.3Hz,3H),0.80(s,3H),0.69(s,3H).¹³C NMR(126MHz,CDCl₃)δ171.70,169.64,162.91,149.24,135.76,130.43,126.78,125.09,124.00,123.23,121.21(2C),119.28,56.87,55.87,49.13,42.34,39.75,38.58,36.35,35.52,34.52,33.79,32.24,31.94,31.81(2C),31.60,28.25,24.21,21.64,21.16,21.02,18.48,17.82,11.95.mp:132-133℃.HRMS(ESI)for C₃₆H₄₉N₃O₂[M+Na]⁺:calcd578.3717,found 578.3704.

Compound Q34, white solid, yield 95％.¹H NMR(500MHz,CDCl₃)δ7.88(d,J＝1.1Hz,1H),7.38(t,J＝11.6Hz,2H),7.10(d,J＝8.1Hz,2H),5.75(dd,J＝5.2,2.2Hz,1H),2.77(d,J＝14.9Hz,1H),2.65(s,3H),2.45–2.36(m,1H),2.30(s,3H),2.27–2.20(m,1H),2.18–2.10(m,2H),2.05(t,J＝6.2Hz,1H),1.96–1.85(m,2H),1.73–1.55(m,4H),1.53(s,3H),1.50–1.40(m,3H),1.35(s,3H),1.32–1.00(m,7H),0.97(d,J＝6.3Hz,3H),0.79(s,3H),0.70(s,3H).¹³C NMR(126MHz,CDCl₃)δ171.71,169.67,162.94,149.22,135.52,133.68,129.44(2C),124.00,121.22,119.87(2C),119.30,56.86,55.86,49.14,42.32,39.75,38.57,36.35,35.53,34.61,33.79,32.25,31.94,31.81,31.66,31.59,28.24,24.21,21.64,21.16,21.02,20.86,18.50,11.94.mp:159-160℃.HRMS(ESI)for C₃₆H₄₉N₃O₂[M+Na]⁺:calcd 578.3717,found 578.3706.

Compound Q35, white solid, yield 95％.¹H NMR(500MHz,CDCl₃)δ10.56(s,1H),8.32(s,1H),7.88(s,1H),5.76(d,J＝2.8Hz,1H),2.75(t,J＝19.7Hz,1H),2.64(s,3H),2.63–2.58(m,1H),2.53–2.41(m,1H),2.19–2.10(m,2H),2.06(d,J＝8.9Hz,1H),1.98–1.84(m,2H),1.73–1.56(m,4H),1.53(s,3H),1.51–1.40(m,2H),1.36(s,3H),1.33–1.04(m,8H),1.00(d,J＝6.0Hz,3H),0.95–0.83(m,1H),0.80(s,3H),0.71(s,3H).¹³C NMR(126MHz,CDCl₃)δ172.09,162.96,161.67,149.23,143.44,139.93(2C),124.03,121.17,119.30,56.84,55.70,49.08,42.38,39.73,38.57,36.36,35.39,33.78,33.17,32.24,31.90,31.81,31.58,30.80,28.26,24.18,21.65,21.14,21.01,18.44,11.94.mp:220-221℃.HRMS(ESI)for C₃₂H₄₃N₅O₄S[M+Na]⁺:calcd 616.2928,found 616.2936.

Compound Q36, white solid, yield 95％.¹H NMR(500MHz,CDCl₃)δ7.87(s,1H),5.75(d,J＝2.9Hz,1H),5.33(d,J＝7.7Hz,1H),4.13–3.98(m,2H),3.98–3.83(m,1H),2.85(d,J＝11.3Hz,2H),2.76(d,J＝14.9Hz,1H),2.64(s,3H),2.25–2.17(m,1H),2.17–2.09(m,2H),2.09–2.01(m,2H),1.89(d,J＝13.0Hz,3H),1.83–1.75(m,2H),1.71–1.56(m,5H),1.52(s,3H),1.45(s,9H),1.35(s,3H),1.31–1.23(m,5H),1.21–1.01(m,6H),0.94(d,J＝6.5Hz,3H),0.79(s,3H),0.68(s,3H).¹³C NMR(126MHz,CDCl₃)δ172.83,169.64,162.91,154.72,149.23,124.00,121.19,119.28,79.67,56.87,55.81,49.12,46.58,42.31(2C),39.73,38.56,36.34,35.50,33.78,33.75,32.23,31.93,31.79,31.77,31.58(2C),28.43(3C),28.24(2C),24.18,21.63,21.14,21.00,18.44,11.91.mp:104-105℃.HRMS(ESI)for C₃₉H₆₀N₄O₄[M+Na]⁺:calcd 671.4507,found671.4548.

Compound Q37, white solid, yield 90％.¹H NMR(400MHz,CDCl₃)δ7.86(s,1H),5.74(d,J＝3.0Hz,1H),3.64(t,J＝8.5Hz,4H),3.60(d,J＝4.0Hz,2H),3.46(d,J＝4.2Hz,2H),2.76(t,J＝11.6Hz,1H),2.63(s,3H),2.42–2.32(m,1H),2.25–2.18(m,1H),2.16–2.08(m,2H),2.03(t,J＝6.0Hz,1H),1.95–1.84(m,1H),1.82–1.73(m,1H),1.71–1.54(m,4H),1.50(s,3H),1.47–1.39(m,1H),1.33(s,3H),1.31–1.00(m,7H),0.95(d,J＝6.4Hz,3H),0.77(s,3H),0.68(s,3H).¹³C NMR(101MHz,CDCl₃)δ172.30,169.61,162.88,149.22,124.00,121.20,119.26,66.96,66.69,56.83,55.80,49.10,46.08,42.30,41.87,39.70,38.56,36.33,35.65,33.78,32.23,31.93,31.81,31.57,31.32,30.10,28.25,24.22,21.63,21.13,21.00,18.55,11.92.mp:155-156℃.HRMS(ESI)for C₃₃H₄₉N₃O₃[M+Na]⁺:calcd 558.3666,found 558.3675.

Compound Q38, white solid, yield 90％.¹H NMR(500MHz,CDCl₃)δ7.87(s,1H),5.75(d,J＝3.0Hz,1H),3.59–3.48(m,2H),3.45–3.30(m,2H),2.74(t,J＝20.9Hz,1H),2.64(s,3H),2.40–2.32(m,1H),2.25–2.16(m,1H),2.16–2.10(m,2H),2.05(d,J＝10.7Hz,1H),1.95–1.85(m,1H),1.80–1.53(m,11H),1.52(s,3H),1.50–1.42(m,2H),1.35(s,3H),1.30–1.03(m,7H),0.96(d,J＝6.5Hz,3H),0.79(s,3H),0.69(s,3H).¹³C NMR(126MHz,CDCl₃)δ171.91,169.60,162.90,149.23,123.99,121.22,119.28,56.85,55.89,49.13,46.75,42.60,42.31,39.72,38.57,36.34,35.76,33.78,32.24,31.94,31.80,31.60,30.50,28.24,26.61,25.59,24.61(2C),24.24,21.63,21.15,21.01,18.55,11.92.mp:159-160℃.HRMS(ESI)for C₃₄H₅₁N₃O₂[M+Na]⁺:calcd556.3874,found 556.3873.

Compound Q39, white solid, yield 90％.¹H NMR(500MHz,CDCl₃)δ7.86(d,J＝9.5Hz,1H),5.74(dd,J＝5.1,2.2Hz,1H),5.41(s,1H),4.03–3.99(m,1H),3.94(dd,J＝8.9,3.0Hz,2H),3.46(td,J＝11.6,1.7Hz,2H),2.75(d,J＝14.9Hz,1H),2.64(s,3H),2.25–2.19(m,1H),2.16–2.10(m,2H),2.07–2.02(m,2H),1.90–1.86(m,2H),1.81-1.75(m,1H),1.72–1.55(m,4H),1.51(s,3H),1.49–1.40(m,4H),1.33(s,3H),1.28–1.22(m,3H),1.19–1.03(m,5H),0.94(d,J＝6.5Hz,3H),0.78(s,3H),0.68(s,3H).¹³C NMR(101MHz,CDCl₃)δ172.88,169.67,162.94,149.22,124.01,121.21,119.30,66.82(2C),56.86,55.80,49.10,45.53,42.30,39.72,38.55,36.33,35.51,33.78,33.74,33.23,32.22,31.92,31.80,31.77,31.57,29.70,28.25,24.19,21.65,21.13,21.00,18.45,11.90.mp:110-111℃.HRMS(ESI)for C₃₄H₅₁N₃O₃[M+Na]⁺:calcd 572.3823,found572.3815.

EXAMPLE 23 preparation of Compound Q43

Compound Q36 (80 mg,0.12 mmol) was placed in a 25mL single-necked flask, methylene chloride (10 mL) was added, trifluoroacetic acid (0.026 mL,0.36 mmol) was added dropwise, stirring was performed at room temperature for 3 hours, TLC was used to detect complete reaction of the starting materials, the solid was directly concentrated, THF (10 mL), DMAP (4 mg,0.024 mmol), acetic anhydride (0.045 mL,0.48 mmol) was added, heating was performed at 60℃for 2 hours, TLC was used to detect complete reaction of the starting materials, cooling was performed at room temperature, water (10 mL) was added, ethyl acetate (10 mL), the extract was separated, and the aqueous phase was extracted again with ethyl acetate (10 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (10 ml×3), saturated sodium chloride solution (10 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=2:1) to give compound Q43 (white solid 50mg, yield 70％).¹H NMR(400MHz,CDCl₃)δ7.85(s,1H),6.95(s,1H),5.76(d,J＝13.8Hz,1H),5.73(s,1H),4.52(d,J＝12.8Hz,1H),3.99(s,1H),3.76(d,J＝13.3Hz,1H),3.14(t,J＝12.7Hz,1H),2.71(dd,J＝26.4,12.7Hz,2H),2.63(s,2H),2.24–2.13(m,2H),2.07(s,3H),2.05–1.97(m,3H),1.88(d,J＝11.6Hz,2H),1.79–1.58(m,4H),1.54(s,1H),1.50(s,3H),1.40(s,2H),1.32(d,J＝5.6Hz,3H),1.30–1.06(m,10H),0.92(d,J＝5.4Hz,3H),0.84(s,1H),0.75(s,2H),0.66(s,3H).¹³C NMR(126MHz,CDCl₃)δ173.70,169.66(2C),162.95,149.20,124.01,121.19,119.31,56.84,56.73,55.74,49.09,46.52,45.40,42.30,42.25,40.84,39.71,38.55,36.34,35.52,33.77,33.60,32.61,32.22,31.92,31.80,31.57,29.70,28.22,24.17,21.65,21.15,21.00,18.40,11.90.mp:140-141℃.HRMS(ESI)for C₃₆H₅₄N₄O₃[M+Na]⁺:calcd 613.4088,found 613.4071.

EXAMPLE 24 preparation of Compound Q44

As in example 19, compound Q7 (500 mg,1.2 mmol), ethyl formate (30 mL), naH (288 mg,12.0 mmol) gave a pale yellow oil Q24.

Then, compound Q24 (previous step oil) was put in a 100mL single-necked flask, ethanol (30 mL), a few drops of water, phenylhydrazine hydrochloride (520 mg,3.6 mmol) and heated at 100 ℃ for 3 hours under reflux, TLC detected complete reaction of the starting materials, water (20 mL) and ethyl acetate (20 mL) were added for extraction and separation, and the aqueous phase was extracted with ethyl acetate (20 ml×3) again, and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=10:1) to give compound Q44 (white solid 570mg, two-step yield 90％).¹³C NMR(126MHz,CDCl₃)δ174.34,151.33,145.72,142.36,137.66,128.93,128.88(2C),128.55(2C),120.74,114.48,60.19,56.91,55.76,49.60,42.31,39.76,38.52,36.96,35.38,33.40,32.27,32.21,31.44,31.42,31.33,31.02,28.17,24.21,21.30,21.04,18.35,14.28,11.90.

EXAMPLE 25 preparation of Compound Q45

In analogy to the synthesis of Q26, compound Q44 (114 mg,0.22 mmol), lithium hydroxide (160 mg,6.6 mmol) and absolute ethanol (10 mL) were reacted at room temperature to give compound Q45 (110 mg as a white solid, yield) 99％).¹H NMR(500MHz,CDCl₃)δ7.47–7.34(m,6H),5.67(d,J＝3.2Hz,1H),2.75(d,J＝14.6Hz,1H),2.42–2.29(m,1H),2.26–2.20(m,1H),2.19(d,J＝14.7Hz,1H),2.15–2.06(m,2H),2.02(d,J＝13.7Hz,1H),1.90–1.75(m,2H),1.71–1.32(m,8H),1.29(d,J＝4.2Hz,3H),1.26–1.16(m,3H),1.13(s,3H),1.11–1.00(m,3H),0.95(s,1H),0.93(s,3H),0.70(s,3H).

EXAMPLE 26 preparation of Compound Q46

In analogy to the synthesis of compound Q37, compound Q45 (110 mg,0.22 mmol) was reacted with EDCI (84.3 mg,0.44 mmol), HOBt (59.4 mg,0.44 mmol), DMAP (119.6 mg,0.66 mmol), morpholine (0.04 mL,0.44 mmol) to give compound Q46 (122.7 mg as a white solid, yield 98％).¹HNMR(400MHz,CDCl₃)δ7.48–7.32(m,6H),5.67(d,J＝3.2Hz,1H),3.64(d,J＝11.8Hz,4H),3.61(d,J＝4.3Hz,2H),3.46(d,J＝4.3Hz,2H),2.75(d,J＝14.6Hz,1H),2.41–2.31(m,1H),2.25–2.15(m,2H),2.15–2.01(m,2H),1.94–1.85(m,1H),1.80(dd,J＝11.8,4.3Hz,1H),1.73–1.31(m,9H),1.29(s,3H),1.26–1.17(m,2H),1.13(s,3H),1.10–1.01(m,2H),0.96(d,J＝6.4Hz,3H),0.93(s,3H),0.68(d,J＝14.2Hz,3H).¹³C NMR(101MHz,CDCl₃)δ172.34,151.31,145.73,142.35,137.65,128.95,128.88(2C),128.57(2C),120.75,114.46,66.97,66.70,56.89,55.80,49.58,46.10,42.33,41.88,39.76,38.51,36.95,35.66,33.40,32.27,32.20,31.43,31.42,31.35,30.10,28.27,24.24,21.30,21.03,18.57,11.92.mp:219-220℃.HRMS(ESI)for C₃₇H₅₁N₃O₂[M+H]⁺:calcd 570.4054,found570.4009.

EXAMPLE 27 preparation of Compound Q47

Compound Q44 (300 mg,0.57 mmol) and NHPI (186.0 mg,1.14 mmol) were placed in a one-necked flask, acetone (20 mL) was added and dissolved, glacial acetic acid (0.033 mL,0.57 mmol) and sodium dichromate (205.2 mg,0.69 mmol) were added, the reaction was kept at 50℃for 2 hours, after TLC detection of the completion of the reaction, the separated liquid was extracted with water (20 mL) and ethyl acetate (20 mL), and the aqueous phase was extracted with ethyl acetate (20 mL. Times.2) again, and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=5:1) to give compound Q47 (white solid 216.3mg, yield 70％).¹³C NMR(126MHz,CDCl₃)δ202.18,175.90,174.18,143.69,141.78,137.66,129.31,128.82(2C),128.62(2C),125.72,113.53,60.18,54.51,50.25,49.65,45.04,43.18,40.47,38.70,37.97,35.27,32.24,31.32,31.18,31.01,30.69,28.45,26.27,21.22,18.62,18.50,14.28,11.97.

EXAMPLE 28 preparation of Compound Q48

Compound Q47 (100 mg,0.18 mmol) was added to a 50mL single-necked flask, 20mL of methanol was added, sodium borohydride (68.4 mg,1.8 mmol) was slowly added under ice-bath, stirred at room temperature for 2 hours, after TLC detection of complete reaction of the starting material, the separated liquid was extracted with water (20 mL) and ethyl acetate (20 mL), the aqueous phase was extracted with ethyl acetate (20 mL. Times.2), and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=3:1) to give compound Q48 (white solid 58.8mg, yield 60％).¹³C NMR(126MHz,CDCl₃)δ174.26,153.97,145.10,142.15,137.65,129.06,128.82(2C),128.63(2C),124.96,114.13,73.60,60.21,56.00,55.13,47.83,42.86,40.52,39.51,38.59,36.84,35.31,33.11,31.97,31.41,31.32,31.02,28.47,26.35,21.07,21.04,18.41,14.28,11.84.

EXAMPLE 29 preparation of Compounds Q49, Q50

Compound Q48 (29 mg,0.053 mmol) and anhydrous tetrahydrofuran (6 mL) were placed in a one-necked flask, the flask was purged with nitrogen, 2M format reagent (0.5 mL,1 mmol) was added dropwise under stirring, the mixture was transferred to room temperature and stirred for 2 hours, after TLC detection of complete reaction of the starting material, water (8 mL) and ethyl acetate (10 mL) were added to extract a liquid, and the aqueous phase was extracted with ethyl acetate (5 mL. Times.2) again, and the organic phases were combined. The organic phase was washed with water (10 ml×3) and saturated sodium chloride solution (8 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=1:1) to give the compound.

Compound Q49, 28mg of white solid, yield 95％.¹HNMR(400MHz,CDCl₃)δ7.49–7.33(m,6H),5.60(d,J＝23.6Hz,1H),3.95(d,J＝5.4Hz,1H),2.76(d,J＝14.6Hz,1H),2.21–2.12(m,1H),2.09–2.02(m,1H),1.95–1.65(m,4H),1.55(d,J＝9.1Hz,2H),1.49(s,1H),1.48–1.42(m,5H),1.41–1.37(m,2H),1.34(s,1H),1.31(s,3H),1.25(s,3H),1.18(s,3H),1.15–1.07(m,2H),1.04(s,1H),0.98(d,J＝6.1Hz,3H),0.94(d,J＝12.2Hz,2H),0.87(s,1H),0.85(s,4H),0.82–0.79(m,1H),0.69(s,3H).¹³C NMR(126MHz,CDCl₃)δ154.11,145.11,142.15,137.67,129.07,128.82(2C),128.64(2C),124.85,114.16,74.72,73.71,56.00,55.10,47.85,42.84,40.58,39.51,38.61,36.86,36.08,34.26,33.11,31.98,31.41,31.18,30.95,29.27,28.60,26.40,21.06,18.92,11.82,7.81,7.72.mp:211-212℃.HRMS(ESI)for C₃₇H₅₄N₂O₂[M+H]⁺:calcd 559.4258,found559.4271.

Compound Q50, 28mg of white solid, yield 95％.¹HNMR(500MHz,CDCl₃)δ7.45–7.33(m,6H),5.91–5.80(m,2H),5.56(d,J＝2.5Hz,1H),5.12(t,J＝13.3Hz,4H),3.94(dd,J＝8.1,2.3Hz,1H),2.75(d,J＝14.7Hz,1H),2.22–2.17(m,4H),2.16(s,1H),2.06(dd,J＝9.5,3.2Hz,1H),2.03(s,1H),1.93–1.88(m,1H),1.86–1.80(m,1H),1.68–1.61(m,2H),1.57–1.34(m,7H),1.30(s,3H),1.24(t,J＝7.2Hz,2H),1.18(s,3H),1.14–1.08(m,2H),0.98(s,3H),0.94(d,J＝6.5Hz,3H),0.70(s,3H).¹³C NMR(101MHz,CDCl₃)δ154.01,145.13,142.13,137.66,133.82(2C),133.79(2C),129.08,128.83(2C),128.64(2C),124.94,118.65,118.58,114.16,73.62,56.01,55.11,47.84,43.83,43.61,42.83,40.53,39.50,38.60,36.85,36.06,35.42,33.11,31.97,31.41,29.27,28.62,26.40,21.05,18.90,11.82.mp:188-189℃.HRMS(ESI)for C₃₉H₅₄N₂O₂[M+H]⁺:calcd 583.4258,found 583.4285.

EXAMPLE 30 preparation of Compound Q51

In analogy to the synthesis of compound Q37, compound Q26 (500 mg,1.18 mmol), EDCI (450.8 mg,2.36 mmol), HOBt (318.6 mg,2.36 mmol), DMAP (783.5 mg,4.72 mmol) and morpholine (0.21 mL,2.36 mmol) reacted to give compound Q51 (112.7 mg as a white solid in yield 90％).¹H NMR(500MHz,CDCl₃)δ7.26(s,1H),5.75(dd,J＝5.2,2.2Hz,1H),3.72–3.65(m,4H),3.65–3.59(m,2H),3.49–3.43(m,2H),2.75(dd,J＝24.6,10.8Hz,1H),2.40–2.34(m,1H),2.25–2.18(m,1H),2.17–2.10(m,2H),2.06–1.87(m,2H),1.81–1.56(m,5H),1.50(s,3H),1.36(d,J＝7.6Hz,3H),1.32–1.03(m,9H),0.97(t,J＝10.7Hz,3H),0.85(s,3H),0.70(s,3H).¹³C NMR(126MHz,CDCl₃)δ172.37,149.89,120.92(2C),112.93(2C),66.97,66.70,56.92,55.80,49.34,46.11,42.31,41.89,39.78,38.86,35.67,35.51,33.35,32.67,32.41,32.08,31.62,31.36,30.13,28.28,24.25,21.29,21.06,18.56,11.91.mp:200-201℃.HRMS(ESI)for C₃₁H₄₇N₃O₂[M+H]⁺:calcd 494.3741,found 494.3741.

Example 31 preparation of Compounds Q52 to Q55

Compound Q51 (50 mg,0.1 mmol), DMAP (3.5 mg,0.02 mmol) were placed in a single-necked flask, THF (10 mL), various anhydrides (0.03 mL,0.3 mmol) were added, the mixture was heated and stirred at 60℃for 2 hours, the reaction was gradually cleared by a visible system, the reaction was checked by TLC, the reaction was returned to room temperature, water (10 mL) and ethyl acetate (10 mL) were added to extract the separated liquid, and the aqueous phase was extracted with ethyl acetate (10 mL. Times.3) and the organic phases were combined. The organic phase was washed with water (10 ml×3) and saturated sodium chloride solution (10 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=2:1) to give the compound.

Compound Q52, white solid 49mg, yield 95％.¹HNMR(400MHz,CDCl₃)δ7.88(dd,J＝12.7,4.9Hz,1H),5.74(dd,J＝5.1,2.1Hz,1H),3.66(dd,J＝9.1,4.5Hz,4H),3.60(d,J＝5.0Hz,2H),3.48–3.42(m,2H),3.10(q,J＝7.4Hz,2H),2.75(d,J＝14.9Hz,1H),2.42–2.29(m,1H),2.27–2.07(m,4H),2.07–1.97(m,1H),1.92–1.81(m,1H),1.82–1.54(m,5H),1.50(s,3H),1.47–1.37(m,2H),1.33(s,3H),1.23(dd,J＝14.1,6.6Hz,4H),1.19–1.02(m,5H),0.95(d,J＝6.5Hz,3H),0.78(s,3H),0.68(s,3H).¹³C NMR(101MHz,CDCl₃)δ173.05,172.39,162.62,149.29,124.08,121.17,118.85,66.95,66.69,56.84,55.79,49.11,46.10,42.31,41.89,39.71,38.57,36.32,35.66,33.77,32.25,31.94,31.88,31.58,31.34,30.11,28.25,27.31,24.22,21.12,21.02,18.54,11.92,8.58.mp:160-161℃.HRMS(ESI)for C₃₄H₅₁N₃O₃[M+Na]⁺:calcd 572.3823,found572.3842.

Compound Q53, 49mg as white solid, yield 95％.¹HNMR(400MHz,CDCl₃)δ7.87(d,J＝1.2Hz,1H),5.74(dd,J＝5.1,2.1Hz,1H),3.66(dd,J＝8.7,4.2Hz,4H),3.61(d,J＝4.9Hz,2H),3.50–3.41(m,2H),3.08–3.01(m,2H),2.75(d,J＝14.9Hz,1H),2.36–2.26(m,1H),2.14–1.90(m,5H),1.89–1.79(m,1H),1.78(dd,J＝14.8,7.4Hz,3H),1.71–1.54(m,4H),1.51(s,3H),1.43–1.33(m,2H),1.34(s,3H),1.28–1.05(m,6H),1.01(t,J＝7.4Hz,3H),0.95(d,J＝6.4Hz,3H),0.78(s,3H),0.68(s,3H).¹³C NMR(101MHz,CDCl₃)δ172.33,172.24,162.57,149.32,124.03,121.16,118.87,66.96,66.70,56.85,55.81,49.11,46.09,42.31,41.88,39.71,38.57,36.32,35.66,35.61,33.77,32.26,31.94,31.88,31.59,31.34,30.11,28.26,24.22,21.13,21.02,18.55,18.05,13.81,11.92.mp:170-171℃.HRMS(ESI)for C₃₅H₅₃N₃O₃[M+Na]⁺:calcd 586.3979,found586.3991.

Compound Q54, 49mg of white solid, yield 95％.¹H NMR(500MHz,CDCl₃)δ7.86(s,1H),5.75(d,J＝3.0Hz,1H),3.91–3.80(m,1H),3.66(t,J＝9.4Hz,4H),3.62(d,J＝4.8Hz,2H),3.47(d,J＝4.4Hz,2H),2.76(d,J＝14.9Hz,1H),2.41–2.32(m,1H),2.26–2.18(m,1H),2.18–2.09(m,2H),2.08–2.00(m,1H),1.95–1.84(m,1H),1.79–1.69(m,1H),1.70–1.55(m,4H),1.52(s,3H),1.50–1.39(m,2H),1.35(s,3H),1.34–1.30(m,1H),1.27(d,J＝6.9Hz,6H),1.23–1.01(m,6H),0.95(d,J＝11.6Hz,3H),0.79(s,3H),0.68(s,3H).¹³C NMR(126MHz,CDCl₃)δ176.20,172.34,162.51,149.32,124.21,121.16,118.83,66.97,66.70,56.86,55.83,49.14,46.11,42.32,41.90,39.73,38.60,36.31,35.67,33.74,32.30,31.95,31.91,31.60,31.35,30.12,28.26,24.22,21.14,21.04,19.35,18.98,18.89,18.55,11.93.mp:198-199℃.HRMS(ESI)for C₃₅H₅₃N₃O₃[M+Na]⁺:calcd 586.3979,found 586.3986.

Compound Q55, white solid 49mg, yield 95％.¹H NMR(500MHz,CDCl₃)δ7.89(s,1H),6.13(s,1H),5.80(s,1H),5.75(d,J＝3.2Hz,1H),3.67(d,J＝3.8Hz,4H),3.62(d,J＝4.5Hz,2H),3.46(s,2H),2.77(dd,J＝14.7,9.6Hz,1H),2.65(s,1H),2.40–2.31(m,1H),2.26–2.20(m,1H),2.16(s,3H),2.12(d,J＝4.8Hz,1H),2.05(d,J＝12.2Hz,1H),1.94–1.87(m,1H),1.79(s,1H),1.68–1.58(m,5H),1.52(s,3H),1.50–1.43(s,2H),1.36(s,3H),1.29–1.16(m,7H),0.96(d,J＝6.3Hz,3H),0.79(s,3H),0.70(s,3H).¹³C NMR(126MHz,CDCl₃)δ172.32,167.15,163.20,149.23,137.87,127.18,125.97,121.22,118.74,66.97,66.70,56.85,55.83,49.12,46.11,42.33,41.90,39.72,38.60,36.35,35.67,33.79,32.21,31.94,31.86,31.60,31.35,30.13,28.26,24.22,21.15,21.04,20.42,18.55,11.93.mp:140-141℃.HRMS(ESI)for C₃₅H₅₁N₃O₃[M+Na]⁺:calcd584.3823,found 584.3815.

EXAMPLE 32 preparation of Compound Q56

In analogy to the synthesis of compound Q49, compound Q25 (50 mg,0.055 mmol), anhydrous tetrahydrofuran (6 mL), 2M ethyl magnesium chloride (0.5 mL,1.1 mmol) was reacted to give compound Q56 (50 mg white solid, yield) 98％).¹HNMR(500MHz,CDCl₃)δ7.26(s,1H),5.75(dd,J＝5.2,2.2Hz,1H),2.72(d,J＝14.5Hz,1H),2.18–2.02(m,3H),1.91–1.84(m,1H),1.74–1.52(m,5H),1.50(s,3H),1.48–1.41(m,5H),1.39(s,1H),1.37(s,3H),1.35–0.98(m,11H),0.94(t,J＝9.3Hz,3H),0.87(d,J＝3.5Hz,2H),0.86(s,1H),0.85(d,J＝3.5Hz,3H),0.84(d,J＝3.7Hz,1H),0.69(s,3H).¹³C NMR(126MHz,CDCl₃)δ149.96,120.90,112.90,74.78,73.24,62.97,56.96,55.75,49.39,42.26,39.79,38.87,36.13,35.50,34.22,33.92,33.34,32.68,32.39,32.11,31.64,31.18,30.94,30.35,29.70,29.26,29.15,28.30,24.25,21.29,21.07,18.85,11.87,10.02,7.80,7.72.mp:210-211℃.HRMS(ESI)for C₃₁H₅₁N₂O[M+H]⁺:calcd 467.3996,found 467.4018.

EXAMPLE 33 preparation of Compound Q57

In a 100mL single-necked flask, morpholine (40 mL) was added and stirred uniformly, and the mixture was heated to reflux at 120℃for 5 hours, after the completion of the reaction of the starting materials by TLC, water (30 mL) and ethyl acetate (30 mL) were added to extract a liquid fraction, and the aqueous phase was extracted with ethyl acetate (20 mL. Times.2) and the organic phase was combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=20:1) to give compound Q57 (360 mg of white solid, yield 80％).¹³C NMR(126MHz,CDCl₃)δ174.74,159.63,151.19,148.86,142.48,141.15,120.73,56.83,55.76,51.48,48.88,43.86,42.38,41.25,39.69,37.65,35.38,33.52,31.89,31.72,31.41,31.05,31.01,28.16,24.17,21.07,20.39,18.31,11.91.

EXAMPLE 34 preparation of Compound Q58

Compound Q57 (360 mg,0.8 mmol) and lithium hydroxide (576 mg,24 mmol) were placed in a 100mL single-necked flask, a mixed solution of aqueous and ethanol was added, stirred at room temperature for 8 hours, TLC was used to detect complete reaction of the starting materials, 2M diluted aqueous hydrochloric acid was added to adjust pH to about 5, ethyl acetate (30 mL) was added, the extract was separated, the aqueous phase was extracted with ethyl acetate (30 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (30 ml×3), saturated sodium chloride solution (30 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=10:1) to give compound Q58 (white solid 349mg, yield 99％).¹H NMR(400MHz,CDCl₃)δ8.45(s,1H),8.28(s,1H),5.79(s,1H),3.09(d,J＝15.6Hz,1H),2.63(d,J＝15.5Hz,1H),2.45–2.35(m,1H),2.33–2.24(m,1H),2.18(d,J＝17.9Hz,1H),2.07(d,J＝12.5Hz,1H),1.95–1.78(m,2H),1.71(dd,J＝28.3,10.8Hz,3H),1.61(s,3H),1.58–1.40(m,3H),1.33(s,3H),1.29–1.19(m,3H),1.19–1.05(m,4H),0.96(d,J＝6.1Hz,3H),0.78(s,3H),0.70(s,3H).mp:265-266℃.HRMS(ESI)for C₂₈H₄₁N₂O₂[M+H]⁺:calcd 437.3163,found437.3190.

EXAMPLE 35 preparation of Compound Q59

In analogy to the synthesis of compound Q37, compound Q58 (349 mg,0.8 mmol), EDCI (306.7 mg,1.6 mmol), HOBt (243.2 mg,1.6 mmol), DMAP (531.2 mg,3.2 mmol) and morpholine (0.14 mL,1.6 mmol) gave compound Q59 (364 mg as a white solid in yield 90％).¹H NMR(500MHz,CDCl₃)δ8.40(s,1H),8.31–8.18(m,1H),5.76(d,J＝2.2Hz,1H),3.65(s,4H),3.59(s,2H),3.45(d,J＝3.1Hz,2H),3.06(d,J＝15.6Hz,1H),2.62(d,J＝15.4Hz,1H),2.39–2.32(m,1H),2.22–2.14(m,2H),2.03(dd,J＝15.7,6.3Hz,1H),1.88(s,1H),1.79–1.63(m,4H),1.59(d,J＝2.7Hz,3H),1.45(dd,J＝20.7,17.4Hz,2H),1.31(s,3H),1.25–1.02(m,7H),1.00–0.89(m,3H),0.86–0.81(m,1H),0.77(s,3H),0.67(d,J＝17.8Hz,3H).¹³C NMR(126MHz,CDCl₃)δ172.29,159.62,151.17,148.85,142.49,141.16,120.73,66.96,66.69,56.82,55.84,48.87,46.09,43.87,42.40,41.88,41.25,39.70,37.65,35.65,33.52,31.89,31.72,31.41,31.33,30.10,28.25,24.19,21.07,20.38,18.54,11.94.mp:151-152℃.HRMS(ESI)for C₃₂H₄₇N₃O₂[M+H]⁺:calcd 506.3741,found 506.3762.

EXAMPLE 36 preparation of Compound Q60

In analogy to the synthesis of compound Q48, compound Q59 (264 mg,0.72 mmol), NHPI (234.7 mg,1.44 mmol), acetone (20 mL), glacial acetic acid (0.042 mL,0.72 mmol), sodium dichromate (256.9 mg,0.864 mmol) reacted to give compound Q60 (262 mg as a white solid, yield 70％).¹³C NMR(126MHz,CDCl₃)δ201.73,173.43,172.25,157.30,149.65,143.15,141.97,125.51,66.97,66.69,54.58,50.27,49.61,46.12,45.16,43.30,42.33,42.17,41.91,39.55,38.69,35.56,32.46,31.38,30.38,30.10,28.57,26.31,21.33,18.71,17.66,12.04.

EXAMPLE 37 preparation of Compound Q61

In analogy to the synthesis of compound Q48, compound Q60 (100 mg,0.18 mmol), 20mL of methanol, sodium borohydride (68.4 mg,1.8 mmol) was reacted to give compound Q61 (white solid 58.8mg, yield) 60％).¹HNMR(500MHz,CDCl₃)δ8.50–8.36(m,1H),8.28(d,J＝1.7Hz,1H),5.71(d,J＝2.4Hz,1H),4.01(d,J＝7.8Hz,1H),3.70–3.63(m,4H),3.61(d,J＝4.8Hz,2H),3.50–3.43(m,2H),3.08(d,J＝15.6Hz,1H),2.61(d,J＝15.6Hz,1H),2.40–2.32(m,1H),2.25–2.18(m,1H),2.07(d,J＝12.8Hz,1H),1.99–1.92(m,1H),1.89–1.85(m,1H),1.82–1.69(m,3H),1.64(s,3H),1.54–1.43(m,4H),1.34(s,3H),1.28–1.21(m,3H),1.14–1.07(m,2H),0.97(d,J＝6.5Hz,3H),0.83(s,3H),0.72(s,3H).¹³C NMR(126MHz,CDCl₃)δ172.27,158.94,151.82,150.66,142.72,141.37,124.78,73.32,66.96,66.69,55.96,55.26,47.56,46.09,43.46,42.97,41.90,41.11,40.60,39.48,37.74,35.63,33.48,31.35(2C),30.14,28.57,26.33,21.15,20.11,18.60,11.90.mp:185-186℃.HRMS(ESI)for C₃₂H₄₇N₃O₃[M+Na]⁺:calcd 544.3510,found 544.3500.

EXAMPLE 38 preparation of Compound Q62

Compound Q7 (300 mg,0.72 mmol) and pyridinium tribromide (278 mg,0.87 mmol) were placed in a 100mL single-necked flask, anhydrous DCM (40 mL) was added, the mixture was stirred overnight at room temperature, after TLC detection of complete reaction of the starting materials, water (30 mL) was added to extract the separated liquid, and the aqueous phase was extracted with dichloromethane (20 mL. Times.2) and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=20:1) to give compound Q62 (white solid 284mg, yield 80％).¹³C NMR(101MHz,CDCl₃)δ209.96,174.74,147.98,120.90,56.62,55.68,51.54,50.42,48.50,44.08,43.74,42.38,39.51,38.86,35.35,31.82,31.56,31.03,30.95,30.84,28.37,28.13,24.10,21.24,19.17,18.30,11.93.

Example 39 preparation of Compound Q63

Compound Q62 (100 mg,0.2 mmol), thiourea (30.4 mg,0.4 mmol) and ethanol (25 mL) were heated under reflux in a single flask for 3 hours, after TLC detection of complete reaction of the starting materials, water (30 mL) and ethyl acetate (30 mL) were added for extraction and separation, and the aqueous phase was extracted with ethyl acetate (20 mL. Times.2) and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=10:1) to give compound Q63 (white solid 56.4mg, yield 60％).¹³C NMR(126MHz,CDCl₃)δ174.75,164.62,151.43,149.07,120.79,115.28,56.92,55.72,51.49,49.38,42.28,39.69,38.80,38.77,35.37,35.18,32.19,32.15,31.57,31.17,31.06,31.02,28.16,24.19,21.45,20.93,18.33,11.87.

EXAMPLE 40 preparation of Compound Q64

In analogy to the synthesis of Q26, compound Q63 (56 mg,0.12 mmol) was hydrolyzed over lithium hydroxide (87 mg,3.6 mmol) to give compound Q64 (50 mg as a white solid, yield) 99％).¹HNMR(400MHz,DMSO)δ11.99(s,1H),6.84(s,2H),5.70(s,1H),2.59(d,J＝15.5Hz,1H),2.27–2.18(m,1H),2.10–2.02(m,1H),2.07(d,J＝21.3Hz,1H),1.97(d,J＝9.9Hz,1H),1.81(s,1H),1.73–1.61(m,2H),1.58(d,J＝15.5Hz,1H),1.52–1.35(m,4H),1.30(s,3H),1.27(d,J＝13.8Hz,1H),1.23(s,3H),1.18(s,2H),1.12–1.02(m,3H),0.97(s,3H),0.89(d,J＝5.5Hz,3H),0.65(s,3H).mp:267-259℃.HRMS(ESI)for C₂₇H₄₀N₂O₂S[M+H]⁺:calcd 457.2883,found 457.2889.

Example 41 preparation of Compounds Q65 to Q66

Compound Q62 (100 mg,0.2 mmol), thioacetamide (30.1 mg,0.4 mmol) and ethanol (25 mL) were heated to reflux in a 50mL single-neck flask for 3 hours, after TLC detection of complete reaction of the starting materials, water (30 mL) and ethyl acetate (30 mL) were added to extract the separated liquid, and the aqueous phase was extracted with ethyl acetate (20 mL. Times.2) again, and the organic phases were combined. The organic phase was washed with water (20 mL. Times.3) and saturated sodium chloride solution (20 mL. Times.2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure to give a white solid Q65, which was used in the next step.

In analogy to the synthesis of Q26, compound Q65, lithium hydroxide (87 mg,3.6 mmol) and aqueous ethanol mixed solution (20 mL) were reacted and hydrolyzed to give compound Q66 (white solid 40mg, two-step yield) 50％).¹H NMR(400MHz,CDCl₃)δ5.76(s,1H),2.83(d,J＝15.3Hz,1H),2.65(s,3H),2.53–2.20(m,3H),2.14(d,J＝17.6Hz,1H),2.04(d,J＝12.1Hz,1H),1.85(d,J＝10.0Hz,2H),1.77–1.52(m,4H),1.49(s,3H),1.40(s,1H),1.36(s,3H),1.32–1.02(m,7H),0.98(s,3H),0.95(d,J＝5.7Hz,3H),0.90–0.72(m,1H),0.70(s,3H).¹³C NMR(101MHz,CDCl₃)δ179.41,162.53,155.82,148.93,125.02,121.08,56.90,55.74,49.34,42.30,39.68,38.93,38.70,35.34,35.24,32.52,32.13,31.57(2C),31.09,30.85,28.15,24.20,21.28,20.98,19.02,18.32,11.89.mp:250-251℃.HRMS(ESI)for C₂₈H₄₁NO₂S[M+H]⁺:calcd456.2931,found 456.2919.

Example 42 preparation of Compound Q67

In analogy to the synthesis of Q26, compound Q7 (1 g,2.41 mmol), lithium hydroxide (1.73 g,72.4 mmol) and aqueous ethanol mixed solution (100 mL) were reacted and hydrolyzed to give compound Q67 (white solid 964mg, yield 99％).¹³C NMR(101MHz,CDCl₃)δ217.14,180.41,149.75,119.91,56.73,55.73,48.85,48.68,42.43,39.71,37.05,35.30,33.71,32.06,31.69,31.19,31.05,30.74,30.23,28.13,27.23,24.15,21.27,19.34,18.28,11.94.

EXAMPLE 43 preparation of Compound Q68

In analogy to the synthesis of compound Q37, compound Q67 (964 mg,2.41 mmol), EDCI (924 mg,4.82 mmol), HOBt (733.4 mg,4.82 mmol), DMAP (1.6 g,9.64 mmol) and morpholine (0.42 mL,4.82 mmol) was reacted to give compound Q68 (1.02 g as a white solid in yield 90％).¹³C NMR(101MHz,CDCl₃)δ216.80,172.27,149.74,119.90,66.94,66.68,56.71,55.83,48.85,48.64,46.07,42.42,41.86,39.70,37.04,35.64,33.70,32.07,31.68,31.30,31.18,30.21,30.10,28.24,27.23,24.17,21.26,19.32,18.52,11.94.

EXAMPLE 44 preparation of Compound Q69

Compound Q68 (100 mg,0.21 mmol) and potassium tert-butoxide (100 mg,1.05 mmol) were dissolved in a 50mL single-necked flask, tert-butanol (10 mL) was added thereto, and isoamyl nitrite (0.13 mL,1.05 mmol) was slowly added dropwise thereto over a water bath at 35℃for thirty minutes, and stirred at room temperature for 5 hours, after which the reaction of the starting materials was detected by TLC, the separated liquid was extracted with water (30 mL) and ethyl acetate (30 mL), and the aqueous phase was extracted with ethyl acetate (20 mL. Times.3) again, and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=2:1) to give compound Q69 (white solid 90mg, yield 86％).¹³C NMR(101MHz,CDCl₃)δ201.71,172.61,153.22,147.89,121.41,66.95,66.69,56.56,55.77,48.65,48.02,46.14,42.37,41.95,39.53,35.95,35.71,35.66,31.65,31.41,31.36,30.52,30.10,28.21,27.16,24.17,21.25,20.95,18.54,11.90.

EXAMPLE 45 preparation of Compound Q70

Compound Q69 (90 mg,0.18 mmol) and hydroxylamine hydrochloride (37.5 mg,0.54 mmol) were added to a 25mL single necked flask, pyridine (5 mL) was added, heated at 100deg.C under reflux for 3 hours, after which TLC detected completion of the reaction, water (30 mL) and ethyl acetate (30 mL) were added to extract fractions, and the aqueous phase was extracted with ethyl acetate (20 mL. Times.3) again, and the organic phases were combined. The organic phase was washed with dilute hydrochloric acid (20 mL), water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=2:1) to give compound Q70 (white solid 83mg, yield) 90％).¹HNMR(400MHz,CDCl₃)δ12.88(s,1H),10.76(s,1H),5.65(s,1H),3.67(t,J＝7.5Hz,4H),3.63(s,2H),3.48(d,J＝3.7Hz,2H),2.38(dd,J＝17.7,7.1Hz,1H),2.28–2.19(m,1H),2.02(d,J＝12.9Hz,2H),1.88(d,J＝17.3Hz,1H),1.73(d,J＝3.3Hz,1H),1.63–1.56(m,2H),1.43(s,3H),1.34–1.28(m,3H),1.21(s,3H),1.17–1.07(m,4H),0.96(d,J＝6.1Hz,2H),0.88–0.85(m,6H),0.84(s,3H),0.82–0.81(m,1H),0.67(s,3H).

EXAMPLE 46 preparation of Compound Q71

Compound Q70 (83 mg,0.162 mmol), potassium hydroxide (2 pieces), ethylene glycol (10 mL), dioxane (5 mL) were heated to reflux in a 50mL single-neck flask at 130℃for 5 hours, after TLC detection of complete reaction of the starting material, water (30 mL) and ethyl acetate (30 mL) were added to extract the separated liquid, and the aqueous phase was extracted with ethyl acetate (20 mL. Times.3) again, and the organic phases were combined. The organic phase was washed with dilute hydrochloric acid (20 mL), water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=2:1) to give compound Q71 (white solid 62.11mg, yield 90％).¹H NMR(400MHz,CDCl₃)δ5.83(d,J＝3.2Hz,1H),3.23(d,J＝15.5Hz,1H),2.45–2.36(m,1H),2.34–2.25(m,2H),2.24–2.01(m,2H),1.97–1.67(m,4H),1.63(s,3H),1.59–1.47(m,3H),1.42(d,J＝12.1Hz,3H),1.38–1.19(m,5H),1.19–1.02(m,3H),0.95(d,J＝6.4Hz,3H),0.79(s,3H),0.70(s,3H).¹³C NMR(101MHz,CDCl₃)δ180.05,159.78,150.69,146.75,122.97,56.65,55.69,48.95,42.26,39.50,38.80,35.30,35.18,33.45,32.35,31.73,31.50,31.34,30.95,30.72,28.11,24.15,21.12,20.91,18.27,11.89.mp:210-211℃.HRMS(ESI)for C₂₆H₃₈N₂O₃[M+Na]⁺:calcd 449.2775,found 449.2778.

EXAMPLE 47 preparation of Compound Q72

In analogy to the synthesis of compound Q27, compound Q71 (42 mg,0.1 mmol), DMAP (3.5 mg,0.02 mmol) and acetic anhydride (0.03 mL,0.3 mmol) were reacted by heating to give compound Q72(45mg,95％).¹H NMR(500MHz,CDCl₃)δ5.83(dd,J＝5.3,2.3Hz,1H),3.67(d,J＝4.3Hz,4H),3.62(d,J＝5.0Hz,2H),3.50–3.43(m,2H),3.23(d,J＝15.5Hz,1H),2.40–2.83(m,1H),2.29(d,J＝15.5Hz,1H),2.26–2.14(m,2H),2.08(dd,J＝9.4,3.3Hz,1H),1.94–1.87(m,1H),1.85–1.75(m,1H),1.74–1.64(m,2H),1.62(s,3H),1.60–1.46(m,3H),1.43(s,3H),1.34(dt,J＝13.5,9.1Hz,2H),1.29–1.20(m,3H),1.20–1.02(m,3H),0.96(d,J＝6.5Hz,3H),0.78(s,3H),0.70(s,3H).¹³C NMR(126MHz,CDCl₃)δ172.25,159.78,150.69,146.71,122.98,66.97,66.70,56.65,55.82,48.95,46.08,42.27,41.89,39.52,38.80,35.65,35.18,33.45,32.35,31.73,31.50,31.34,31.29,30.11,28.22,24.18,21.12,20.91,18.53,11.91.mp:250-251℃.HRMS(ESI)for C₃₀H₄₅N₃O₃[M+Na]⁺:calcd518.3352,found 518.3359.

EXAMPLE 48 preparation of Compounds Q73-Q74

Compound Q68 (50 mg,0.11 mmol) was placed in a 50mL single-necked flask, anhydrous tetrahydrofuran (20 mL) was added to the solution, naH (28.8 mg,1.2 mmol) was added, diethyl oxalate (0.03 mL,0.22 mmol) was added under nitrogen protection, stirring was performed at room temperature for 20 minutes, the visible system turned orange-yellow, TLC detection of the starting material was complete, water (10 mL) and ethyl acetate (10 mL) were added to extract the fractions, the aqueous phase was extracted once more with ethyl acetate (10 mL), and the organic phases were combined. The organic phase was washed with water (10 ml×3), saturated sodium chloride solution (10 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure to give a pale yellow solid Q73, which was unstable and was immediately followed.

Compound Q73 (previous solid) was added to a 50mL single-necked flask, acetic acid (20 mL), potassium acetate (22 mg,0.22 mmol), hydrazine hydrate (0.18 mL,3.6 mmol), heated to reflux at 130℃for 5 hours, TLC was checked for complete reaction, 2M sodium hydroxide solution (60 mL) was added and stirred for 10 minutes, saturated aqueous sodium bicarbonate solution (30 mL), 20mL ethyl acetate, the extract was separated, the aqueous phase was extracted again with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure followed by silica gel column chromatography (PE: ea=2:1) to give compound Q74 (white solid 56mg, two-step yield 90％).¹HNMR(500MHz,CDCl₃)δ5.75(dd,J＝5.2,2.2Hz,1H),4.44–4.27(m,2H),3.70–3.63(m,4H),3.61(d,J＝5.2Hz,2H),3.51–3.42(m,2H),3.19(d,J＝15.7Hz,1H),2.43–2.29(m,1H),2.26–2.16(m,2H),2.13(dt,J＝17.7,5.4Hz,1H),2.04(dd,J＝9.8,2.8Hz,1H),1.92–1.82(m,1H),1.81–1.74(m,1H),1.71–1.54(m,4H),1.51(s,3H),1.50–1.40(m,2H),1.39(s,1H),1.37(s,2H),1.36(s,3H),1.33–1.01(m,7H),0.95(d,J＝6.5Hz,3H),0.85(s,3H),0.69(s,3H).¹³C NMR(126MHz,CDCl₃)δ172.42,161.30,149.21(2C),121.07(2C),117.66,66.97,66.70,60.69,56.90,55.79,49.37,46.12,42.31,41.91,39.75,38.48,35.67,35.66,33.20,32.91,32.35,32.05,31.59,31.37,30.12,28.26,24.24,21.42,21.07,18.56,14.38,11.92.mp:144-145℃.HRMS(ESI)for C₃₄H₅₁N₃O₄[M+Na]⁺:calcd588.3772,found 588.3798.

EXAMPLE 49 preparation of Compound Q75

In analogy to the synthesis of compound Q73 in example 48, compound Q68 (50 mg,0.11 mmol), naH (28.8 mg,1.2 mmol) and diethyl oxalate (0.03 mL,0.22 mmol) were reacted to give light yellow solid Q73.

Compound Q73 (previous solid) and hydroxylamine hydrochloride (23 mg,0.33 mmol) were added to a 50mL single-necked flask, an aqueous ethanol mixture (20 mL) was added, the mixture was heated to reflux at 100℃for 3 hours, the reaction was returned to room temperature by TLC, water (20 mL) and ethyl acetate (20 mL) were added to extract the separated liquid, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=2:1) to give the compound Q75(42mg,90％).¹H NMR(500MHz,CDCl₃)δ5.83–5.67(m,1H),4.48–4.33(m,2H),3.71–3.62(m,4H),3.59(d,J＝4.7Hz,2H),3.45(d,J＝4.4Hz,2H),2.97(t,J＝17.1Hz,1H),2.38–2.31(m,1H),2.24–2.19(m,1H),2.16(dd,J＝13.9,5.0Hz,1H),2.10(dd,J＝14.7,9.3Hz,1H),2.02(d,J＝12.5Hz,1H),1.91–1.83(m,1H),1.81–1.74(m,1H),1.73–1.49(m,6H),1.47(d,J＝9.4Hz,3H),1.39(dd,J＝8.2,6.0Hz,5H),1.34–1.26(m,3H),1.24–0.98(m,7H),0.94(d,J＝3.5Hz,1H),0.93(s,3H),0.68(s,3H).¹³C NMR(101MHz,CDCl₃)δ175.71,172.27,160.90,153.61,147.52,122.23,110.80,66.93,66.67,61.69,56.76,55.74,49.36,46.06,42.25,41.86,39.57,38.60,36.98,35.60,32.33,31.96,31.46,31.29,30.50,30.04,29.82,28.21,24.20,21.72,20.93,18.54,14.18,11.87.mp:140-141℃.HRMS(ESI)for C₃₄H₅₀N₂O₅[M+Na]⁺:calcd 589.3612,found 589.3638.

EXAMPLE 50 preparation of Compound Q76

As in example 19, compound Q7 (1.00 g,2.41 mmol), ethyl formate (30 mL), naH (578 mg,24.1 mmol) gave Q24 as a pale yellow oil.

Compound Q24 (previous step oil) and hydroxylamine hydrochloride (502 mg,7.23 mmol) were added to a 100mL single-necked flask, an aqueous ethanol mixture (50 mL) was added, the mixture was heated to reflux at 100℃for 3 hours, TLC checked for complete reaction, the mixture was returned to room temperature, the solution was extracted with water (30 mL) and ethyl acetate (30 mL), the aqueous phase was extracted with ethyl acetate (30 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (30 ml×3), saturated sodium chloride solution (30 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=2:1) to give the compound Q76(952mg,90％).¹H NMR(500MHz,CDCl₃)δ8.02(s,1H),5.77(d,J＝3.4Hz,1H),3.84(s,1H),3.66(s,3H),2.57(d,J＝14.9Hz,1H),2.39–2.31(m,1H),2.27–2.19(m,1H),2.14–2.10(m,1H),2.05(d,J＝12.6Hz,1H),1.92–1.85(m,1H),1.83–1.75(m,1H),1.67–1.61(m,3H),1.54(dd,J＝10.7,6.7Hz,2H),1.48(s,3H),1.39(s,3H),1.34–1.30(m,2H),1.26(d,J＝8.4Hz,3H),1.18(s,1H),0.94(s,3H),0.93(s,3H),0.79(s,1H),0.69(s,3H),0.68(s,1H).

EXAMPLE 51 preparation of Compounds Q77 to Q79

In analogy to the synthesis of compound Q48, compound Q76 (250 mg,0.57 mmol), NHPI (186.0 mg,1.14 mmol), 20mL acetone, glacial acetic acid (0.033 mL,0.57 mmol), sodium dichromate (205.2 mg,0.69 mmol) was reacted to give compound Q77 (182 mg, 70% yield as white solid).

In analogy to the synthesis of compound Q49, compound Q77 (82 mg,0.18 mmol), 20mL of methanol, sodium borohydride (68.4 mg,1.8 mmol) was reacted and reduced to give compound Q78 (white solid 50mg, 60% yield).

In analogy to the synthesis of compound Q49, compound Q78 (50 mg,0.11 mmol), anhydrous tetrahydrofuran (6 mL), 2M ethyl magnesium chloride reagent (1 mL,2.2 mmol) reacted to give the compound Q79.¹HNMR(400MHz,CDCl₃)δ8.06(d,J＝24.1Hz,1H),5.70(t,J＝10.3Hz,1H),3.97(t,J＝10.7Hz,1H),2.59(d,J＝15.0Hz,1H),2.12(d,J＝15.1Hz,1H),2.04(d,J＝5.3Hz,1H),1.93(dd,J＝12.1,7.8Hz,1H),1.87–1.80(m,1H),1.68–1.62(m,2H),1.60–1.55(m,2H),1.53(s,3H),1.50(s,1H),1.48–1.43(m,5H),1.42(s,3H),1.39–1.34(m,2H),1.34–1.28(m,2H),1.26(d,J＝7.2Hz,3H),1.20(d,J＝4.6Hz,1H),1.19–1.13(m,2H),1.11–1.03(m,2H),1.01(s,3H),0.96(d,J＝6.4Hz,3H),0.86(d,J＝2.4Hz,3H),0.72(s,3H).¹³C NMR(101MHz,CDCl₃)δ172.01,151.16,149.63,125.94,108.56,74.73,73.38,55.95,55.09,47.79,42.80,40.71,39.41,38.97,36.68,36.06,34.24,32.04,31.18,30.95,30.31,29.91,29.26,28.55,26.38,21.47,21.04,18.89,11.78,7.80,7.71.mp:191-192℃.HRMS(ESI)for C₃₁H₄₉NO₃[M+H]⁺:calcd 484.3785,found 484.3811.

Example 52 preparation of Compounds Q80-Q81

In analogy to the synthesis of Q24, compound Q68 (100 mg,0.21 mmol), 20mL of ethyl formate, naH (51 mg,2.1 mmol) reacted to give Q80 as a pale yellow oil.

Compound Q80 (previous step oil) was added to a 50mL single neck flask, 20mL of ethanol, a few drops of water, o-fluorophenylhydrazine hydrochloride (80 mg,0.63 mmol), heated to reflux at 100 ℃ for 3 hours, TLC detected complete reaction of the starting material, 10mL of water, 20mL of ethyl acetate, the extract was separated, the aqueous phase was extracted with ethyl acetate (20 ml×3), and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure followed by silica gel column chromatography (PE: ea=10:1) to give compound Q81 (white solid 111mg, two-step yield 90％).¹H NMR(500MHz,CDCl₃)δ7.52–7.37(m,3H),7.21(dd,J＝16.1,8.4Hz,2H),5.68(d,J＝3.4Hz,1H),3.71–3.63(m,4H),3.61(d,J＝5.0Hz,2H),3.50–3.41(m,2H),2.75(d,J＝14.6Hz,1H),2.41–2.31(m,1H),2.26–2.14(m,2H),2.14–2.02(m,2H),1.94–1.83(m,1H),1.83–1.75(m,1H),1.71–1.41(m,6H),1.41–0.99(m,14H),0.97(d,J＝3.8Hz,2H),0.95(d,J＝6.1Hz,3H),0.70(s,3H).¹³C NMR(126MHz,CDCl₃)δ172.34(2C),138.68(2C),131.19,131.12,123.99,120.80(3C),114.67(2C),66.96,66.69,56.88,55.80,49.58(2C),46.10,42.32,41.89,39.75,38.53,36.73,35.66,33.37,32.19,31.44(2C),31.35(2C),30.11,28.27,24.23,21.03,18.56,11.92.mp:250-251℃.HRMS(ESI)for C₃₇H₅₀FN₃O₂[M+H]⁺:calcd 588.3960,found 588.3952.

EXAMPLE 53 preparation of Compound Q82

Compound Q6 (1 g,2.35 mmol), palladium on carbon (500 mg), 30mL ethyl acetate were placed in an autoclave and stirred at room temperature for 24 hours under 4 mpa, TLC detected complete reaction of the starting materials, suction filtered, the filter cake was washed with ethyl acetate (10 ml×3), and directly concentrated followed by silica gel column chromatography (PE: ea=10:1) to give compound Q82 (white solid 1g, two-step yield 99％).¹³C NMR(126MHz,CDCl₃)δ174.36,79.09,60.17,56.58,56.34,55.84,54.72,42.50,39.88,38.94,37.36,36.48,35.35,35.07,32.89,31.32,31.00,28.14(2C),27.62,24.19,21.29,20.63,18.27,15.50,14.37,14.26,12.02.

EXAMPLE 54 preparation of Compounds Q83-Q84

In analogy to the synthesis of Q24-Q25, compound Q82 (1.00 g,2.41 mmol), 30mL of ethyl formate, naH (578 mg,24.1 mmol) was reacted to give Q83 as a pale yellow oil.

Then, compound Q83 (former oily substance), 30mL of glacial acetic acid, potassium acetate (473 mg,4.82 mmol) and 85% hydrazine hydrate (0.42 mL,8.51 mmol) were heated to reflux to obtain compound Q84 (white solid 1g, yield) 91.8％).¹³C NMR(126MHz,CDCl₃)δ174.37,149.38,134.09,112.57,60.19,56.47,55.88,54.97,52.85,42.41,39.95,37.90,35.61,35.36,34.76,33.40,32.56,31.44,31.33,31.00,28.14,24.39,24.27,22.40,20.85,18.30,14.27,13.72,11.94.

EXAMPLE 55 preparation of Compounds Q85 to Q86

In analogy to the synthesis of Q26-Q27, compound Q84 (1 g,2.21 mmol) was hydrolyzed over lithium hydroxide (1.60 g,66.4 mmol) to give compound Q85 (white solid 1.03g, 99% yield). Thereafter, compound Q85 (1.03 g,2.21 mmol), DMAP (73.37 mg,0.44 mmol) and acetic anhydride (0.63 mL,6.63 mmol) were reacted to give Compound Q86 (white solid 927mg, yield) 90％).¹H NMR(500MHz,CDCl₃)δ7.84(d,J＝5.6Hz,1H),2.71–2.66(m,1H),2.64(s,3H),2.45–2.35(m,1H),2.30–2.21(m,1H),2.08–1.99(m,2H),1.87–1.75(m,3H),1.72–1.66(m,1H),1.66–1.57(m,2H),1.51–1.33(m,8H),1.29(s,3H),1.24(s,3H),1.19–1.05(m,4H),0.94(d,J＝6.5Hz,3H),0.89–0.83(m,2H),0.79(s,3H),0.67(s,3H).¹³C NMR(126MHz,CDCl₃)δ180.06,169.65,162.87,124.41,119.37,56.43,55.84,54.74,52.77,42.40,39.89,37.49,35.60,35.32,34.67,34.55,32.48,31.63,30.97,30.75,28.12,24.95,24.24,22.69,21.61,20.88,18.26,13.70,11.95.mp:135-136℃.HRMS(ESI)for C₂₉H₄₄N₂O₃[M+Na]⁺:calcd491.3244,found 491.3243.

EXAMPLE 56 preparation of Compound Q87

In analogy to the synthesis of compound Q37, compound Q86 (145 mg,0.31 mmol), EDCI (120.8 mg,0.63 mmol), HOBt (85.2 mg,0.63 mmol), DMAP (154.0 mg,1.26 mmol) and morpholine (0.59 mmol) were reacted to give compound Q87 (140 mg as a white solid in yield 95％).¹H NMR(500MHz,CDCl₃)δ7.81(s,1H),3.64(s,4H),3.59(s,2H),3.45(s,2H),2.65(d,J＝10.8Hz,1H),2.62(s,3H),2.39–2.30(m,1H),2.19–2.08(m,1H),2.05–1.94(m,2H),1.88–1.74(m,3H),1.68(d,J＝14.9Hz,1H),1.63–1.54(m,2H),1.52–1.32(m,7H),1.28(s,3H),1.22(s,3H),1.16–1.02(m,4H),0.93(d,J＝6.7Hz,3H),0.88–0.81(m,2H),0.77(s,3H),0.67–0.62(s,3H).¹³C NMR(126MHz,CDCl₃)δ172.32,169.59,162.83,124.38,119.34,66.95,66.69,56.42,55.93,54.74,52.75,46.09,42.40,41.88,39.89,37.48,35.65,35.58,34.66,34.53,32.48,31.63,31.32,30.10,28.22,24.95,24.26,22.67,21.60,20.87,18.51,13.69,11.96.mp:135-136℃.HRMS(ESI)for C₃₃H₅₁N₃O₃[M+Na]⁺:calcd 560.3823,found 560.3817.

Example 57 preparation of Compounds Q88 to Q89

In analogy to the synthesis of Q24-Q25, compound Q19 (1.00 g,2.41 mmol), 30mL of ethyl formate, naH (578 mg,24.1 mmol) was reacted to give Q88 as a pale yellow oil.

Then, compound Q88 (former oily substance), 30mL of glacial acetic acid, potassium acetate (473 mg,4.82m mol) and 85% hydrazine hydrate (0.42 mL,8.51 mmol) were heated to obtain Compound Q89 (white solid 1g, yield) 91.8％).¹³CNMR(101MHz,CDCl₃)δ174.00,150.02(2C),120.87(2C),112.81,60.20,56.95,55.78,49.38,42.25,39.78,38.87,35.52,35.40,34.81,33.33,32.71,32.45,32.11,31.63,28.23,24.23,21.56,21.31,21.06,18.63,14.29,11.86.

EXAMPLE 58 preparation of Compounds Q90-Q92

In analogy to the synthesis of Q26, compound Q89 (104 mg,0.221 mmol) was hydrolyzed together with lithium hydroxide (160 mg,6.64 mmol) to give compound Q90 (103 mg as a white solid, 99% yield).

In analogy to the synthesis of compound Q37, compound Q90 (100 mg,0.23 mmol), EDCI (90 mg,0.46 mmol), HOBt (62 mg,0.46 mmol), DMAP (152.7 mg,0.92 mmol) and morpholine (0.04 mL,0.46 mmol) were reacted and condensed to give compound Q91 (111 mg as a white solid, 95% yield).

Compound Q91 (111 mg,0.22 mmol) and DMAP (7.337 mg,0.044 mmol) were added to a 50mL single neck flask, 20mL of THF and isobutyric anhydride (0.063 mL,0.663 mmol) were added, the mixture was heated and stirred at 60℃for 2 hours, the visible system was gradually cleared, the reaction of the starting materials was detected by TLC and was complete, the reaction was returned to room temperature, 10mL of water, 20mL of ethyl acetate were added, the aqueous phase was extracted with ethyl acetate (20 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (20 ml×3), saturated sodium chloride solution (20 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=2:1) to give compound Q92 (white solid 114mg, yield 90％).¹HNMR(500MHz,CDCl₃)δ7.88–7.85(m,1H),5.75(dd,J＝5.2,2.2Hz,1H),3.92–3.81(m,1H),3.66(d,J＝4.4Hz,4H),3.62(d,J＝5.2Hz,2H),3.49–3.44(m,2H),2.77(d,J＝14.9Hz,1H),2.33–2.22(m,2H),2.17–2.10(m,2H),2.04(dd,J＝9.4,3.2Hz,1H),1.88–1.81(m,1H),1.75–1.56(m,5H),1.52(s,3H),1.50–1.39(m,4H),1.35(s,3H),1.29–1.25(m,6H),1.22–1.01(m,7H),0.95(d,J＝6.5Hz,3H),0.90–0.82(m,1H),0.79(s,3H),0.69(s,3H).¹³C NMR(126MHz,CDCl₃)δ176.22,172.03,162.55,149.35,124.23,121.17,118.89,66.97,66.70,56.88,55.75,49.15,46.09,42.26,41.92,39.71,38.60,36.31,35.74,35.66,33.75,33.59,32.28,31.97,31.94,31.91,31.60,28.28,24.21,21.78,21.14,21.04,19.35,18.98,18.68,11.89.mp:96-97℃.HRMS(ESI)for C₃₆H₅₅N₃O₃[M+Na]⁺:calcd 600.4136,found 600.4138.

EXAMPLE 59 preparation of Compound Q93

In analogy to the synthesis of compound Q27, compound Q90 (200 mg,0.46 mmol), DMAP (15.16 mg,0.091 mmol), 20mL THF, acetic anhydride (0.14 mL,1.37 mmol) was reacted by heating to give compound Q93 (199 mg as a white solid in yield) 90％).¹³C NMR(126MHz,CDCl₃)δ179.87,169.68,162.90,149.30,124.05,121.18,119.25,56.90,55.87,49.18,42.28,39.75,38.57,36.34,35.59,35.47,35.02,33.77,32.26,31.98,31.83,31.60,28.25,24.24,21.63,21.57,21.16,21.02,18.63,11.90.

EXAMPLE 60 preparation of Compounds Q94 to Q95

Compound Q93 (110 mg,0.23 mmol), EDCI (90 mg,0.46 mmol), HOBt (62 mg,0.46 mmol), DMAP (152.7 mg,0.92 mmol), N-Boc-4-aminopiperidine (92 mg,0.46 mmol) were placed in a 25mL single-neck flask, anhydrous DCM (10 mL) was added, stirring at room temperature was performed for 12 hours, after completion of the reaction of the starting materials by TLC, 20mL of water was added, the aqueous phase was extracted with DCM (10 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (10 mL), saturated NaCl solution (10 mL), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography to give Compound Q94 (white solid 145mg, yield 95%).

Compound Q94 (145 mg,0.22 mmol) was placed in a 50mL single-necked flask, 15mL of methylene chloride was added, trifluoroacetic acid (0.048 mL,0.66 mmol) was added dropwise, stirring was carried out at room temperature for 3 hours, TLC was used to detect complete reaction of the starting materials, the solid was directly concentrated, then 15mL of THF, DMAP (7.3 mg,0.044 mmol) and acetic anhydride (0.083 mL,0.88 mmol) were added, heating was carried out at 60℃for 2 hours, TLC was used to detect complete reaction of the starting materials, cooling was carried out to room temperature, 10mL of water was added, 10mL of ethyl acetate was added, the extract was separated, and the aqueous phase was extracted again with ethyl acetate (10 mL. Times.3), and the organic phases were combined. The organic phase was washed with water (10 ml×3), saturated sodium chloride solution (10 ml×2), dried over anhydrous Na ₂SO₄, and concentrated under reduced pressure, followed by silica gel column chromatography (PE: ea=2:1) to give compound Q95 (white solid 93mg, yield 70％).¹H NMR(500MHz,CDCl₃)δ7.87(s,1H),5.75(dd,J＝5.2,2.2Hz,1H),5.38(d,J＝18.1Hz,1H),4.53(d,J＝19.7Hz,1H),4.03–3.94(m,1H),3.76(d,J＝18.1Hz,1H),3.20–3.07(m,1H),2.79–2.74(m,1H),2.74–2.67(m,2H),2.64(s,3H),2.17–2.10(m,4H),2.09(s,3H),2.06–2.02(m,2H),1.91(dd,J＝16.7,9.4Hz,1H),1.85–1.77(m,1H),1.74–1.55(m,6H),1.52(s,3H),1.48–1.37(m,4H),1.35(s,3H),1.31–1.00(m,15H),0.94(d,J＝6.5Hz,3H),0.78(s,3H),0.68(s,3H).¹³C NMR(126MHz,CDCl₃)δ172.56,168.92,162.92,149.25(2C),124.01,121.20,119.30,56.86,55.73,49.12,46.50,45.31,42.25,40.56,39.70,38.57,37.28,36.34,35.58,35.48,33.78,32.84,32.22,31.95,31.80,31.58,29.71,28.28,24.20,22.27,21.65,21.46,21.13,21.01,18.67,11.88.mp:110-111℃.HRMS(ESI)for C₃₇H₅₆N₄O₃[M+Na]⁺:calcd 627.4245,found 627.4245.

EXAMPLE 61 preparation of Compound Q96

In analogy to the synthesis of compound Q37, compound Q93 (110 mg,0.23 mmol), EDCI (90 mg,0.46 mmol), HOBt (62 mg,0.46 mmol), DMAP (152.7 mg,0.92 mmol) and morpholine (0.04 mL,0.46 mmol) were reacted to give compound Q96 (120 mg as a white solid in yield 95％).¹HNMR(500MHz,CDCl₃)δ7.86(d,J＝1.1Hz,1H),5.73(dd,J＝5.2,2.3Hz,1H),3.64(d,J＝4.1Hz,4H),3.60(d,J＝5.2Hz,2H),3.48–3.39(m,2H),2.75(d,J＝14.9Hz,1H),2.63(s,3H),2.32–2.19(m,2H),2.16–2.07(m,2H),2.05–1.99(m,1H),1.83–1.75(m,1H),1.73–1.54(m,5H),1.50(s,3H),1.47–1.36(m,3H),1.33(s,3H),1.27–1.01(m,8H),0.94(d,J＝6.5Hz,3H),0.77(s,3H),0.67(s,3H).¹³C NMR(126MHz,CDCl₃)δ171.90,169.59,162.90,149.26,124.01,121.20,119.29,66.96,66.69,56.86,55.74,49.13,46.05,42.25,41.88,39.70,38.56,36.32,35.73,35.64,33.78,33.57,32.21,31.94,31.79,31.58,28.27,24.20,21.74,21.62,21.13,21.00,18.68,11.88.mp:90-91℃.HRMS(ESI)for C₃₄H₅₁N₃O₃[M+Na]⁺:calcd 572.3823,found 572.3822.

EXAMPLE 62 preparation of Compounds Q97-Q98

In analogy to the synthesis of Q24-Q25, compound Q23 (1.07 g,2.41 mmol), 30mL of ethyl formate, naH (578 mg,24.1 mmol) was reacted to give a pale yellow oil Q97.

Then, compound Q97 (former oily substance), 30mL of glacial acetic acid, potassium acetate (473 mg,4.82 mmol) and 85% hydrazine hydrate (0.42 mL,8.51 mmol) were heated and refluxed to obtain compound Q98 (white solid 1.03g, yield) 91.8％).¹³C NMR(126MHz,CDCl₃)δ173.96,150.01(2C),120.89(2C),112.87,60.18,56.96,56.01,49.40,42.25,39.81,38.88,35.61,35.55,35.50,34.47,33.35,32.70,32.42,32.12,31.64,28.28,25.64,25.45,24.24,21.30,21.07,18.66,14.27,11.89.

EXAMPLE 62 preparation of Compound Q99

In analogy to the synthesis of Q26, compound Q98 (103 mg,0.221 mmol) was hydrolyzed together with lithium hydroxide (160 mg,6.64 mmol) to give compound Q99 (103 mg as a white solid, yield) 99％).¹H NMR(500MHz,MeOD)δ7.90(s,1H),5.97–5.85(m,1H),2.91(d,J＝15.0Hz,1H),2.31–2.10(m,5H),1.93–1.86(m,1H),1.78–1.59(m,5H),1.57(s,3H),1.55–1.50(m,1H),1.44(s,4H),1.42–1.39(m,1H),1.38–

1.04(m,11H),0.97(d,J＝6.5Hz,3H),0.90(s,3H),0.79–0.71(m,3H).

EXAMPLE 63 preparation of Compound Q100

In analogy to the synthesis of compound Q27, compound Q99 (100 mg,0.23 mmol), DMAP (7.8 mg,0.046 mmol), 20mL THF, acetic anhydride (0.07 mL,0.69 mmol) was reacted by heating to give compound Q100 (102 mg of white solid, yield) 90％).¹³C NMR(101MHz,CDCl₃)δ179.55,169.68,162.98,149.24,124.05,121.24,119.35,56.87,56.01,49.15,42.26,39.73,38.58,36.34,35.61,35.52,34.08,33.78,32.23,31.97,31.81,31.59,28.28,25.61,25.15,24.22,21.64,21.15,21.01,18.65,11.90.

EXAMPLE 64 preparation of Compound Q101

In analogy to the synthesis of compound Q37, compound Q100 (102 mg,0.21 mmol), EDCI (80 mg,0.42 mmol), HOBt (57 mg,0.42 mmol), DMAP (139.4 mg,0.84 mmol) and morpholine (0.04 mL,0.42 mmol) were condensed to give compound Q101 (112 mg as a white solid in yield 95％).¹HNMR(500MHz,CDCl₃)δ7.86(d,J＝4.8Hz,1H),5.74(dd,J＝5.2,2.3Hz,1H),3.68–3.64(m,4H),3.61(d,J＝5.1Hz,2H),3.51–3.41(m,2H),2.76(d,J＝14.9Hz,1H),2.64(s,3H),2.30(td,J＝15.2,8.8,5.5Hz,2H),2.18–

2.09(m,2H),2.08–2.01(m,1H),1.88–1.81(m,1H),1.75–1.55(m,6H),1.51(s,3H),1.48–

1.38(m,3H),1.34(s,3H),1.28–1.00(m,9H),0.92(d,J＝6.5Hz,3H),0.78(s,3H),0.67(s,3H).¹³C NMR(126MHz,CDCl₃)δ171.92,169.64,162.93,149.26,124.01,121.23,119.31,66.98,66.71,56.89,56.04,49.16,46.09,42.26,41.88,39.74,38.58,36.34,35.71,35.63,33.78,33.22,32.23,31.97,31.80,31.59,28.30,26.00,25.73,24.21,21.63,21.15,21.01,18.68,11.90.mp:105-106℃.HRMS(ESI)for C₃₅H₅₃N₃O₃[M+Na]⁺:calcd 586.3979,found 586.3981.

Example 65 preparation of Compounds Q102-Q106

In analogy to the synthesis of Q24-Q26, compound Q13 (400 mg,1.0 mmol), 30mL of ethyl formate, naH (240 mg,10.0 mmol) reacted to give a pale yellow oil Q102.

Then, compound Q102 (previous step oil), 20mL glacial acetic acid, potassium acetate (196.2 mg,2.0 mmol), 85% hydrazine hydrate (0.15 mL,3.0 mmol) were heated to reflux to afford compound Q103 (386 mg as a white solid, 91% yield).

Hydrolysis of compound Q103 (3836 mg,0.91 mmol), lithium hydroxide (65mg, 27.3 mmol) gave compound Q104 (380 mg, 99% yield as a white solid).

In analogy to the synthesis of compound Q92, compound Q104 (380 mg,0.91 mmol), DMAP (30 mg,0.182 mmol), 30mL THF, isobutyric anhydride (0.37 mL,3.64 mmol) was reacted by heating to give compound Q105 (393 mg, 90% yield as white solid).

In analogy to the synthesis of compound Q37, compound Q105 (393 mg,0.82 mmol), EDCI (312 mg,1.64 mmol), HOBt (222 mg,1.64 mmol), DMAP (543.7 mg,3.28 mmol) and morpholine (0.16 mL,1.64 mmol) were reacted to give compound Q106 (427.7 mg as a white solid in yield 95％).¹H NMR(500MHz,CDCl₃)δ7.86(d,J＝4.4Hz,1H),5.74(dd,J＝5.2,2.2Hz,1H),3.91–3.80(m,1H),3.71–3.57(m,6H),3.48(s,2H),2.76(d,J＝15Hz,1H),2.39(d,J＝11.9Hz,1H),2.18–2.10(m,2H),2.09–1.97(m,3H),1.89–1.80(m,1H),1.73–1.55(m,4H),1.51(s,3H),1.49–1.38(m,1H),1.35(s,3H),1.34–1.30(m,1H),1.27(d,J＝1.7Hz,3H),1.26(d,J＝1.7Hz,3H),1.22–1.03(m,5H),1.00(d,J＝6.0Hz,3H),0.79(s,3H),0.74(s,3H).¹³C NMR(126MHz,CDCl₃)δ176.17,171.53,162.45,149.39,124.22,121.06,118.78,67.04,66.72,56.92,56.46,49.11(2C),46.34,42.48,41.96,39.66,39.62,38.59,36.31,33.86,33.74,32.29,31.94,31.91,31.59,28.39,24.19,21.11,21.03,19.62,19.34,18.98,11.96.mp:230-231℃.HRMS(ESI)for C₃₄H₅₁N₃O₃[M+Na]⁺:calcd 572.3823,found 572.3825.

EXAMPLE 66 preparation of Compounds Q107 to Q111

Synthesis of Compounds Q107 to Q111 Using Q9 as starting Material, obtained by a procedure analogous to the Synthesis of Compounds Q102 to Q106 .¹H NMR(500MHz,CDCl₃)δ7.86(s,1H),5.74(dd,J＝5.1,2.1Hz,1H),3.91–3.80(m,1H),3.72–3.50(m,8H),2.76(d,J＝14.9Hz,1H),2.72–2.68(m,1H),2.17–2.08(m,2H),2.04–

1.95(m,1H),1.88–1.80(m,2H),1.76–1.54(m,5H),1.51(s,3H),1.49–1.36(m,2H),1.35(s,3H),1.27(s,3H),1.26(s,3H),1.20(dd,J＝11.0,3.9Hz,1H),1.16(d,J＝6.7Hz,3H),1.13–1.08(m,2H),0.79(s,3H),0.73(s,3H).¹³C NMR(126MHz,CDCl₃)δ176.19,175.36,162.47,149.32,124.23,121.10,118.79,67.12,66.89,56.30,52.78,49.05,46.32,42.18,42.07,39.54(2C),38.58,36.31,33.76,32.26,31.91(2C),31.87,31.59,27.87,24.29,21.08,21.06,19.35,18.96,17.42,12.30.mp:248-249℃.HRMS(ESI)for C₃₃H₄₉N₃O₃[M+Na]⁺:calcd 558.3666,found 558.3670. Example 67: activity test of cholic acid derivative for degrading HMGCR-GFP

The following method was used to determine the HMGCR degrading activity of the compounds prepared in the examples of the present invention using the CHO-7/HMGCR-GFP cell line. The CHO-7/HMGCR-GFP cell line expressed HMGCR-GFP fusion protein by stable transfection in Chinese hamster ovary epithelial cells CHO-7.

The experimental procedure is briefly described as follows: CHO-7/HMGCR-GFP cells were cultured in DMEM/F12 (1:1) medium (containing 5% fbs,100units/ML PENICILLIN-Streptomycin) and, when cell confluency reached 80%, digested and blown up with 0.25% pancreatin (containing EDTA) and seeded at 3 x10 ⁴ cells per well into 24 well plates. After incubation for 24 hours at 37℃in a 5% CO ₂ incubator, the medium was aspirated and washed 1 pass with PBS buffer (137mM NaCl,2.7mM KCl,10mM Na ₂HPO₄,2mM KH₂PO₄). Then, the culture medium of the apolipoprotein DMEM/F12 (containing 5% of the apolipoprotein serum, 1. Mu.M of lovastatin, 10. Mu.M of mevalonic acid, 100units/ML PENICILLIN-Streptomycin) prepared by the examples of the invention, which is equipped with different concentrations, is added, so that the accumulation of HMGCR can be induced to the greatest extent under the action of statin. After 16 hours of incubation, the medium was aspirated, 150 μl of cell lysate Reporter Lysis Buffer (Promega, E397A) was added to each well, the 24 well plate was frozen in a-80℃refrigerator, thawed 1 more time at room temperature, and then shaken on a vortex for 10 minutes at 800rpm to achieve sufficient lysis of the cells.

100 Μl of the lysate was pipetted into a 96-well black microplate (Perkinelmer, 6005329), and the fluorescence intensity of the green fluorescent protein EGFP was measured with a EnVsion microplate reader (Perkinelmer) at 485nm excitation light and 535nm emission light. Compounds prepared according to the examples of the present invention were normalized to 100% in the DMSO untreated group using a concentration gradient of 0.003. Mu.M, 0.01. Mu.M, 0.03. Mu.M, 0.1. Mu.M, 0.3. Mu.M, 1. Mu.M, 3. Mu.M, 10. Mu.M, and half-maximal inhibitory concentration EC50 was calculated using GRAPHPAD PRISM software. The activity of the HMGCR is measured by the above method, and the degradation effect is shown in Table 1.

TABLE 1 Activity data of Compounds prepared in accordance with the examples of the present invention to degrade HMGCR-GFP

As can be seen from the data in table 1, the compounds prepared in the examples of the present invention all have a remarkable effect on the degradation of HMGCR, wherein the EC50 of the most active compounds Q37 and Q54 are 0.49 μm and 0.45 μm, respectively.

EXAMPLE 68 experiments of degradation of HMGCR by Compounds prepared according to the examples of the present invention

The following method was used to determine the degradation of HMGCR by the compounds prepared according to the examples of the present invention. The experimental procedure is briefly described as follows: CHO-7 cells were transferred to a 60mm dish at 6×10 ⁵ and cultured for 1 day. The medium was aspirated, washed 1 time with PBS, PBS was removed, and the medium (5% deproteinized and deproteinized serum, 1. Mu.M lovastatin, 10. Mu.M mevalonic acid, 100units/ML PENICILLIN-Streptomycin) containing the compound prepared in the examples of the present invention at various concentrations was added and treated for 16 hours. Cells in the petri dish were scraped off with a cell scraper, transferred to a 15ml centrifuge tube, and placed on ice. After centrifugation at 1000g for 5min at 4℃the medium was decanted, 1ml PBS buffer was added and transferred to a 1.5ml centrifuge tube and centrifuged at 1000g for 5min at 4 ℃. The PBS was removed and 300 mu L SDS LYSIS Buffer (10 mM Tris-HCl (pH 7.6), 100mM sodium chloride, 1% sodium dodecyl sulfate and protease inhibitor) was added 15 times through a 7 gauge needle. Centrifuge at 13200rpm for 10 min at room temperature, transfer 270. Mu.l of the supernatant to a fresh 1.5ml centrifuge tube, add 90. Mu.l of 4x Loading Buffer (1.2% sodium dodecyl sulfate, 30% glycerol, 150mM Tris-HCl (pH 6.8), 0.6% beta-mercaptoethanol and appropriate amount of bromophenol blue), mix well, cook for 10 min at 95℃and store in a-20℃refrigerator.

Detection of degradation of HMGCR by Western Blot hybridization (Western Blot)

1) Loading: a30. Mu.l sample of each protein was loaded into wells of an 8% SDS-PAGE gel.

2) Electrophoresis: the protein was allowed to enter the gel at 85 volts for 30 minutes followed by 120 volts for 1 hour.

3) Transferring: after electrophoresis, the gel was taken out and put into a membrane transfer device in the order of filter paper, nitrocellulose membrane, gel and filter paper, and the membrane was transferred at a constant voltage of 100 v for 1.5 hours with attention to bubble removal.

4) Closing: after the transfer, nitrocellulose membranes were immersed in TBST buffer (20 mM Tris-HCl (pH 7.4), 150mM sodium chloride and 0.05% Tween-20) containing 5% skimmed milk powder, and incubated for 1 hour at room temperature in a shaker.

5) Incubation resistance: after the completion of the blocking, the membrane was washed 3 times with TBST buffer. Mu.g/ml of HMGCR antibody (IgG-A9, mouse monoclonal antibody recognizing hamster HMGCR) was added and incubated at room temperature for 2 hours or overnight at 4 ℃. Antibodies were recovered and membranes were washed 4 times for 8 minutes each with TBST.

6) Secondary antibody incubation: TBST was discarded, goat anti-mouse HRP conjugated secondary antibody was added at 1:5000 dilution, and incubated for 1 hour at room temperature. TBST was washed 4 times for 8 minutes each.

7) Color development and photographing: ECL luminescent substrate was covered on nitrocellulose membrane, sealed with preservative film, and pictures were taken with LAS4000 luminescence/bioluminescence image analyzer (Fujifilm).

8) Quantitative analysis of HMGCR protein the resulting grey scale pictures were quantitatively analyzed with Image J (NIH) software to 100% of the non-dosed group.

The measurement results are shown in fig. 2 (a and c): the compound prepared by the embodiment of the invention can obviously degrade HMGCR. Fig. 2 (b and d) shows the quantitative analysis results: the compound prepared in the embodiment of the invention has very strong HMGCR degradation activity, and the EC50 values of Q37 and Q54 are respectively 0.22 mu M and 0.26 mu M.

Example 69: intracellular cholesterol accumulation assay

The experimental procedure is briefly described as follows: CHO-7/HMGCR-GFP cells were cultured in DMEM/F12 (1:1) medium (containing 5% fbs,100units/ML PENICILLIN-Streptomycin) and, when cell confluency reached 80%, digested and blown up with 0.25% pancreatin (containing EDTA) and seeded at 3 x 10 ⁴ cells per well into 12 well plates. After incubation for 24 hours at 37℃in a 5% CO ₂ incubator, the medium was aspirated and washed 1 pass with PBS buffer (137mM NaCl,2.7mM KCl,10mM Na ₂HPO₄,2mM KH₂PO₄).

Then, 1ml of a deproteinized DMEM/F12 medium (containing 10% deproteinized serum, 1. Mu.M lovastatin, 10. Mu.M mevalonic acid, 100units/ML PENICILLIN-Streptomycin) prepared with the compound prepared in the present invention at a concentration of 0. Mu.M, 1. Mu.M and 10. Mu.M was added, and after 16 hours of incubation in a 5% CO ₂ incubator at 37 ℃, the medium was aspirated, washed 1 time with PBS buffer (137mM NaCl,2.7mM KCl,10mM Na ₂HPO₄,2mM KH₂PO₄), fixed with 4% Paraformaldehyde (PFA) for 30 minutes, washed 3 times with PBS buffer (137mM NaCl,2.7mM KCl,10mM Na ₂HPO₄,2mM KH₂PO₄), and incubated at room temperature in a dark place for 30 minutes by diluting ethanol-dissolved Filipin mother liquor (5 mg/ml) with PBS containing 10% FBS to a final concentration of 50. Mu.g/ml. Then washed 3 times with PBS buffer (137mM NaCl,2.7mM KCl,10mMNa ₂HPO₄,2mM KH₂PO₄) and 2 times with double distilled water. The chips were sealed, dried overnight, and then placed under a microscope (Zeiss Z2 wide-field microscope (non-confocal) DAPI channel) to observe the accumulation of intracellular cholesterol, and as shown in fig. 3, it was detected that the intracellular cholesterol was completely free from accumulation after treatment with compounds Q37 and Q46.

The cholic acid derivative provided by the invention has potential drug research value in preventing and/or treating hypercholesterolemia, hypertriglyceridemia, atherosclerosis and nonalcoholic steatohepatitis as a compound for reducing cholesterol and triglyceride in a body, and provides a new thought for searching a novel drug for preventing and/or treating hypercholesterolemia, hypertriglyceridemia, atherosclerosis and nonalcoholic steatohepatitis.

The protection of the present invention is not limited to the above embodiments. Variations and advantages that would occur to one skilled in the art are included in the invention without departing from the spirit and scope of the inventive concept, and the scope of the invention is defined by the appended claims.

Claims

1. A preparation method of cholic acid derivative is characterized in that Q1 is taken as a starting material, and compound Q7 is obtained through TBS protection, 4-dimethyl, deprotection, oxidation, witting reaction and reduction; the preparation method is shown in a route (1):

2. A preparation method of cholic acid derivative is characterized in that a compound Q7 is taken as a starting material, and is subjected to glycol protection, reduction, iodo, cyano substitution, hydrolysis and esterification to obtain Q19; the preparation method is shown in a route (2):

3. A preparation method of cholic acid derivatives is characterized in that compounds Q7 are used as starting materials, and cyclized, hydrolyzed, acylated, oxidized and reduced to obtain cholic acid derivatives shown as formulas Q58, Q59 and Q61; the preparation method is shown in a route (3):

4. A preparation method of cholic acid derivatives is characterized in that a compound Q7 is taken as a starting material, and the cholic acid derivatives Q64 and Q66 can be respectively obtained through bromination, cyclization and hydrolysis; the preparation method is shown in a route (4):

5. A preparation method of cholic acid derivative is characterized in that compound Q7 is taken as a starting material, and cholic acid derivative Q68 is obtained through hydrolysis and acylation; the preparation method is shown in a route (5):

6. A preparation method of cholic acid derivatives is characterized in that Q68 is taken as a starting material, and cholic acid derivatives Q71 and Q72 are obtained through condensation, cyclization and acylation; the preparation method is shown in a route (6):

7. A preparation method of cholic acid derivatives is characterized in that a compound Q68 is taken as a starting material, and cholic acid derivatives Q74 and Q75 are obtained through ester condensation and cyclization respectively; the preparation method is shown in a route (7):

8. a preparation method of cholic acid derivative is characterized in that compound Q68 is taken as a starting material, and cholic acid derivative Q81 is obtained through ester condensation and cyclization; the preparation method is shown in a route (8):

9. A preparation method of cholic acid derivative is characterized in that compound Q5 is taken as a starting material, and compound Q9 is obtained through oxidation and esterification; the preparation method is shown in a route (9):

10. A preparation method of cholic acid derivative is characterized in that compound Q4 is taken as a starting material, and is subjected to iodo, cyano substitution, hydrolysis and esterification to obtain compound Q13; the preparation method is shown in a route (10):

and/or the number of the groups of groups,

Taking a compound Q15 as a raw material, and obtaining Q23 through oxidation, witting reaction, glycol protection removal and reduction; the preparation method is shown in a route (11):

and/or the number of the groups of groups,

Taking a compound Q36 as a starting material, and obtaining cholic acid derivative Q43 through hydrolysis and condensation; the preparation method is shown in a route (12):

and/or the number of the groups of groups,

Using a compound Q24 as a starting material, and performing addition cyclization, hydrolysis and acylation to obtain compounds Q45 and Q46; the preparation method is shown in a route (13):