CN101007796A - Quinary-heterocyclic compound, its preparation method and medical uses - Google Patents

Abstract

The present invention relates to a compound of general formula I, including its racemate or optical isomer, or a pharmaceutically acceptable salt or solvate thereof, and the definition of each substituent group in formula I is as described in the description. The present invention also relates to pharmaceutical compositions comprising said compounds of general formula I. The present invention also relates to the preparation method of the compounds of general formula I and the application of these compounds in the preparation of weight loss and antineoplastic drugs.

Description

Novel five-membered heterocyclic compound and its preparation method and medical application

technical field

The present invention relates to some novel five-membered heterocyclic compounds and their preparation methods, pharmaceutical compositions containing the above-mentioned compounds, and the preparation of such compounds for the treatment and/or prevention of obesity, non-insulin-dependent diabetes mellitus and its complications, and coronary heart disease , hypertension, hyperlipidemia and other obesity complications, as well as the use of drugs for tumors and their complications.

Background technique

Obesity is a multifactorial chronic disease affected by biological behavior and environmental factors. When the human body eats more calories than it consumes, the excess calories are stored in the body in the form of fat, and the body mass index (BMI) increases due to the increase in body fat. BMI is defined as: BMI=weight (kg)/height ² (m ² ), and those with a BMI greater than 24 are obese patients. With the development of the economy and the improvement of people's living standards, in recent years, the number of obese patients in the world has doubled every five years. At present, there are at least 250 million obese patients in the world, accounting for about 7% of the total population ( Claude B., The New England Journal Of Medicine, 2000, 343:1888-1889). Obesity not only affects the appearance, but may also cause coronary heart disease, hypertension, hyperlipidemia, non-insulin-dependent diabetes and certain malignant tumors (Must A., et al., J.Am.Med.Assos, 1999, 282: 1523 ). Therefore, it has very important clinical significance for the prevention and treatment of obesity.

At present, the drugs for the treatment of obesity are mainly divided into three categories according to their mechanism of action. The first category is appetite suppressants, the representative drug is sibutramine (Sibutramine), which is a central food inhibitor, inhibits the reuptake of norepinephrine and serotonin, makes people feel full, reduces appetite, Eat less. Its common side effects include dry mouth, nausea, anorexia, abdominal pain, constipation, insomnia, etc., and it can slightly increase blood pressure and heart rate. The second category is digestion and absorption blockers, and the representative drug is orlistat (Olistat), which blocks fat from being broken down into small molecules of triglycerides by inhibiting pancreatic lipase. Its common side effects include causing gastrointestinal dysfunction in patients and reducing the absorption of fat-soluble vitamins in patients. The third category is metabolic stimulants, such as thyroxine and β3 receptor agonists (Zhou Yuzhao, Huang Zhongyi, Chinese Pharmacy, 2000, 11(4), 168-169).

Recent studies have shown that targeting fatty acid synthase (FAS) can achieve the goal of weight loss by inhibiting the synthesis of fatty acids and suppressing appetite. At the same time, it can also improve non-insulin-dependent diabetes, reduce high blood pressure, coronary embolism and Incidence of other obesity complications (Loftus TM, Jaworsky DE, Frehywot GL, Townsend CA, Ronnett GV, Lane MD, Kuhajda FP, Reduced food intake and bodyweight in mice treated with fatty acid synthase inhibitors, Science, 2000, 288: 2379 -81). Studies on the mechanism of action of specific inhibitors of fatty acid synthase have shown that fatty acid synthase inhibitors can reduce the synthesis of fatty acids. Due to the blockage of fatty acid synthesis, the concentration of its substrate malonyl-CoA (malonyl-CoA) increases, and malonyl coenzyme A (malonyl-CoA) increases. Acyl-CoA can directly act on the feeding center of the hypothalamus, inhibiting the secretion of neuropeptide Y (NPY) that promotes feeding, leading to feeding inhibition. On the other hand, in peripheral tissues such as liver and adipose tissue, fatty acid synthase inhibitors can increase the activity of carnitine palmitoyltransferase-1 (O-carnitine palmitoyltransferase-1, CPT-1), thereby enhancing the metabolism of fatty acids. Oxidation and energy consumption. Pharmacological experiments have shown that specific inhibitors of fatty acid synthase have less toxic and side effects, and the suppression of appetite can produce its own feedback regulation (Thupari JN, Landree LE, Ronnett GV, Kuhajda FP, C75 increase peripheral energy utilization and fatty acid oxidation indiet-induced obesity, Proc Natl Acad Sci, USA, 2002, 99:9498-502).

Summary of the invention

The purpose of the present invention is to find and develop small molecule inhibitors acting on fatty acid synthase (FAS), by which it inhibits fatty acid synthase, on the one hand reduces the synthesis and enrichment of fatty acids; on the one hand can increase the substrate malonyl-CoA The concentration directly acts on the feeding center of the hypothalamus, inhibits eating, and achieves the goal of weight loss through compensatory consumption of excessive body fat. At the same time, it can also improve non-insulin-dependent diabetes and reduce the incidence of hypertension, coronary embolism and other obesity complications. Since the FAS enzyme is highly expressed in certain tumor tissues, such as colon cancer, prostate cancer, ovarian cancer, breast cancer, etc., the compound can be further used as an anticancer drug. In addition, the compound is applied to raising livestock and poultry, which can reduce the fat content in their meat.

The present inventors have now found that compounds of the general formula I described below can achieve the above objects.

Therefore, in one aspect, the present invention provides a compound of general formula I, its racemate or optical isomer, or a pharmaceutically acceptable salt or solvate thereof.

In another aspect, the present invention provides a pharmaceutical composition comprising at least one compound of general formula I, or its racemate or optical isomer, or a pharmaceutically acceptable salt or solvate thereof, and at least one A pharmaceutically acceptable carrier, diluent or excipient.

In another aspect, the present invention also relates to a method for preparing the compound of general formula I or a pharmaceutically acceptable salt or solvate thereof.

On the other hand, the present invention also relates to the compound of formula I, or its racemate or optical isomer, or a pharmaceutically acceptable salt or solvate thereof for the prevention and/or treatment of obesity and the resulting Use in medicines resulting in various diseases.

In yet another aspect, the present invention also relates to the compound of formula I, or its racemate or optical isomer, or a pharmaceutically acceptable salt or solvate thereof in the preparation for the prevention and/or treatment of fatty acid synthase Use in the medicine of various diseases such as colon cancer, prostate cancer, ovarian cancer, breast cancer etc. that overexpresses.

In yet another aspect, the present invention also relates to the compound of formula I, or its racemate or optical isomer, or a pharmaceutically acceptable salt or solvate thereof in poultry and livestock of low-fat and high-lean meat type Applications.

Detailed description of the invention

In one aspect, the present invention provides a compound of general formula I, including its racemate or optical isomer, or a pharmaceutically acceptable salt or solvate thereof,

in:

R ₁ is a C ₃ to C ₁₈ straight chain or branched chain alkyl group, or a C ₃ to C ₁₈ straight chain or branched chain alkenyl group, and the alkyl or alkenyl group optionally has a group selected from halogen, nitrile, C ₁ -C ₆ straight-chain or branched alkoxy, C ₂ -C ₆ straight-chain or branched alkenyloxy, trifluoromethoxy, phenoxy, benzyloxy, C ₃ -C ₆ cycloalkyl, A substituent of a 5-6 membered aromatic carbocyclic or heterocyclic group, the aromatic heterocyclic ring includes 1 to 4 heteroatoms selected from O, S and N; there may be no substitution on the aromatic carbocyclic or heterocyclic ring, It can also be substituted by 1 to 5 substituents selected from the following: halogen, nitro, hydroxyl, hydroxymethyl, trifluoromethyl, trifluoromethoxy, C ₁ to C ₆ straight or branched chain alkyl, C ₂ ~C ₆ straight chain or branched alkenyl, C ₁ ~C ₄ alkoxy, C ₂ ~C ₄ alkenyloxy, phenoxy, benzyloxy, nitrile, carboxyl and amino;

R ₂ is selected from H, C ₁ ~ C ₆ linear or branched alkyl, trifluoromethyl, benzyl, and benzyl with substituents selected from the following group on the benzene ring: halogen, nitro, Hydroxy, hydroxymethyl, trifluoromethyl, trifluoromethoxy, C ₁ ~C ₆ straight chain or branched chain alkyl, C ₂ ~C ₆ straight chain or branched chain alkenyl, C ₁ ~C ₄ alkoxy group, C ₂ ~C ₄ alkenyloxy, phenoxy, benzyloxy, nitrile, carboxyl and amino;

R ₃ is =CH ₂ , =CZ ₂ , or =CHZ, wherein Z is a C ₁ to C ₃ linear or branched alkyl group, F, Cl or Br;

X is selected from O and S atoms;

Y is selected from halogen, C ₁ ~C ₄ straight chain or branched chain alkyl, C ₂ ~C ₄ straight chain or branched chain alkenyl, C ₁ ~C ₄ alkoxy, C ₂ ~C ₄ alkenyloxy, nitrile radical, nitro, hydroxyl, hydroxymethyl, trifluoromethyl, trifluoromethoxy, and amino.

According to a preferred embodiment, the compound of the present invention is preferably a compound represented by general formula II, including its racemate or optical isomer, or a pharmaceutically acceptable salt or solvate thereof,

in:

R ₁ is a C ₃ to C ₁₈ straight chain or branched chain alkyl group, or a C ₃ to C ₁₈ straight chain or branched chain alkenyl group, and the alkyl or alkenyl group optionally has a group selected from halogen, nitrile, C ₁ -C ₆ straight-chain or branched alkoxy, C ₂ -C ₆ straight-chain or branched alkenyloxy, trifluoromethoxy, phenoxy, benzyloxy, C ₃ -C ₆ cycloalkyl , a substituent of a 5- to 6-membered aromatic carbocyclic or heterocyclic group, wherein the aromatic heterocyclic ring includes 1 to 4 heteroatoms selected from O, S and N; there may be no substitution on the aromatic carbocyclic or heterocyclic ring, It can also be substituted by 1 to 5 substituents selected from the following: halogen, nitro, hydroxyl, hydroxymethyl, trifluoromethyl, trifluoromethoxy, C ₁ to C ₆ straight or branched chain alkyl, C ₂ ~C ₆ straight chain or branched alkenyl, C ₁ ~C ₄ alkoxy, C ₂ ~C ₄ alkenyloxy, phenoxy, benzyloxy, nitrile, carboxyl and amino;

X is selected from O and S atoms.

Particularly preferred compounds of the invention include:

trans-2-n-hexyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

trans-2-n-heptyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

trans-2-n-octyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

trans-2-n-nonyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

trans-2-n-decyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

trans-2-n-undecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

trans-2-n-dodecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

trans-2-n-tridecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

trans-2-(dec-9-enyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

trans-2-(cis-non-3-enyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

trans-2-[3-(Benzyloxy)-propyl]-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

cis-2-n-hexyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

cis-2-n-heptyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

cis-2-n-octyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

cis-2-n-nonyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

cis-2-n-decyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

cis-2-n-undecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

cis-2-n-dodecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

cis-2-n-tridecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

cis-2-(dec-9-enyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

cis-2-(cis-non-3-enyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

cis-2-[3-(Benzyloxy)-propyl]-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid;

And its pharmaceutically acceptable salt or solvate.

On the other hand, the present invention also relates to a method for preparing a compound of general formula I and/or general formula II, or a pharmaceutically acceptable salt thereof, or a solvate thereof.

According to a specific embodiment, the present invention provides a process for the preparation of a compound of formula Ia,

The method comprises the steps of:

1) making itaconic anhydride (III) react with p-methoxybenzyl alcohol (IV),

The compound of formula V is obtained,

V

2) reacting the compound of formula V with methyl iodide (CH ₃ I) in the presence of a base (such as lithium diisopropylamide (LDA)) to obtain the compound of formula VI,

3) react the compound of formula VI with the aldehyde R ₁ CHO in the presence of a base (such as: lithium diisopropylamide (LDA)), wherein the definition of R ₁ is the same as the definition in the compound of the above-mentioned general formula I, to obtain the compound of formula Ia, and

4) Optionally, column separation to obtain trans and cis compounds of formula Ia,

Trans-I cis-I.

According to another specific embodiment, the present invention also provides a method for preparing a compound of formula IIa, comprising hydrolyzing the racemic compound of formula Ia with, for example, trifluoroacetic acid to obtain a racemic compound of formula IIa, or converting trans or The cis formula Ia compound is hydrolyzed with trifluoroacetic acid, for example, to obtain the trans or cis formula IIa compound, respectively,

  trans-IIa             cis-IIa

Wherein, R ₁ is as defined above.

The above-mentioned preparation method is specifically described as follows:

Reaction step one:

Itaconic anhydride (III) and p-methoxybenzyl alcohol (IV) were reacted at 55-60°C for 40 hours. After the reaction mixture was cooled to room temperature, diethyl ether was added thereto, and then the diethyl ether solution was poured into NaHCO ₃ aqueous solution, the water layer was separated, adjusted to pH ~ 3 with dilute hydrochloric acid, and a solid was precipitated. Filter and recrystallize from ethyl acetate-n-hexane to obtain the compound of formula V as a white solid.

Reaction step two:

At -78°C, lithium diisopropylamide (LDA) was used as a base, and the compound of formula V was reacted with methyl iodide. After 2 hours, 1.5 mol/L dilute sulfuric acid solution was added to terminate the reaction. The reactant was extracted with ether, and the ether layer was washed with saturated NaCl aqueous solution until neutral, and dried with anhydrous sulfuric acid. After filtration, it was spin-dried and separated by column (n-hexane/ethyl acetate/glacial acetic acid) to obtain the compound of formula VI.

Reaction step three:

Add the compound of formula VI dropwise to the anhydrous tetrahydrofuran solution of LDA at -78°C. After 2 hours, add the anhydrous tetrahydrofuran solution of aldehyde (R ₁ CHO) dropwise at -78°C. Keep the reaction at low temperature for 6 hours and then add 1.5mol/ L of sulfuric acid solution to terminate the reaction. The reaction mixture was extracted with ether, and the ether layer was washed with saturated NaCl aqueous solution until neutral, and dried with anhydrous sulfuric acid. After filtration, it was spin-dried and separated by column (ethyl acetate/n-hexane) to obtain trans and cis compounds of formula Ia.

Reaction step four:

The trans and cis compounds of Formula Ia obtained above were hydrolyzed with trifluoroacetic acid respectively, and after reacting overnight, they were extracted with NaHCO ₃ aqueous solution, and the aqueous layer was acidified and then extracted with diethyl ether. The ether layer was dried and concentrated, and the residue was separated by column to obtain trans and cis compounds of Formula IIa, respectively.

Other compounds of formula I or II can be synthesized according to established methods in the field of organic synthesis by changing raw materials and reagents, or by further converting the compound of formula Ia or IIa.

Those skilled in the art can understand that asymmetric synthesis can be used to obtain a single optical isomer. However, the resolution of racemates is the main means to obtain optically pure compounds. There are mainly four resolution methods: crystallization, chromatography, kinetics and enzymatic methods. For the resolution of the compound racemate involved in the present invention, the crystallization method with practical value is preferred: add a kind of chiral base ( Resolving agent) to form diastereoisomers, utilizing the different solubility of diastereomers in solvents to make one of them preferentially precipitate. The preferred chiral base can be ephedrine and the like. Chromatography mainly uses HPLC chiral columns for separation to obtain compounds of single optical purity.

Those skilled in the art will appreciate that the compounds of the present invention may also be used in the form of their pharmaceutically acceptable salts or solvates. The pharmaceutically acceptable salts of the compounds of formula I or II include conventional salts formed from pharmaceutically acceptable inorganic or organic acids or bases and acid addition salts of quaternary ammonium. Examples of suitable salts with acids include hydrochloric, hydrobromic, sulfuric, phosphoric, nitric, perchloric, fumaric, acetic, propionic, succinic, glycolic, formic, lactic, maleic, tartaric acids , citric acid, pamoic acid, malonic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, naphthalene-2-sulfonic acid, benzene Salts of sulfonic acid, hydroxynaphthoic acid, hydroiodic acid, malic acid, steroic acid, tannic acid, etc. Other acids, such as oxalic acid, although not pharmaceutically acceptable in themselves, can be used in the preparation of salts used as intermediates to obtain the compounds of the invention and their pharmaceutically acceptable salts. Examples of suitable salts with bases include sodium, lithium, potassium, magnesium, aluminum, calcium, zinc, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethyl Salts of diamines, N-methylglucamine and procaine. When referring to the compounds of the present invention below, the compounds of formula I or formula II and their pharmaceutically acceptable salts and solvates are included.

The present invention also includes prodrugs of the compounds of the present invention, which, upon administration, are chemically transformed by metabolic processes and then become active drugs. Typically, such prodrugs are functional derivatives of the compounds of the invention which are readily converted in vivo to the desired compound of formula (I). General methods for selecting and preparing suitable prodrug derivatives are described, for example, in "Design Of Prodrugs", H Bund Saard, Elsevier ed., 1985.

The present invention also includes active metabolites of the compounds of the present invention.

Another aspect of the present invention relates to pharmaceutical compositions comprising a compound of the present invention and at least one pharmaceutically acceptable carrier, diluent or excipient, which are useful for in vivo therapy and are biocompatible. The pharmaceutical composition can be prepared in various forms according to different administration routes.

The pharmaceutical composition of the present invention comprises an effective dose of the compound of formula I of the present invention or a pharmaceutically acceptable salt or solvate such as hydrate, and one or more suitable pharmaceutically acceptable carriers, diluents or excipients. The pharmaceutical carriers here include but are not limited to: ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human albumin, buffer substances such as phosphate, glycerol, sorbic acid, potassium sorbate, saturated vegetable Partial glyceride mixtures of fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose Substance, macrogol, sodium carboxymethylcellulose, polyacrylates, beeswax, lanolin.

The compounds of the present invention are a class of fatty acid synthase inhibitors. Compared with the compound C75 mentioned in US Pat. No. 5,981,575, the compound of the present invention not only has better fatty acid synthase inhibitory activity but also has good stability, and is especially suitable for industrial production and long-term storage. The compound of the present invention can be used for but not limited to treating obesity, type II diabetes and its complications, coronary heart disease, hypertension and hyperlipidemia and other obesity complications.

The compound of the present invention can also be extended to be used in the preparation of drugs for anti-tumor and its complications. Because certain tumor cells have high FAS expression, the compounds involved in the present invention have certain inhibitory or cytotoxic effects on certain tumor cells. Therefore, the compounds of formula I and/or formula II of the present invention or their pharmaceutically acceptable salts or solvates can be used for relieving or treating tumors such as colon cancer, prostate cancer, ovarian cancer, breast cancer and their symptoms. In addition, the compound of the present invention can be used to reduce the fat level of chickens, ducks and other poultry and pigs and other livestock, so it can be used to raise low-fat and high-lean poultry and livestock. The dosage regimen of the compounds of the invention may vary according to the use involved.

Pharmaceutical compositions comprising the compounds of this invention may be administered in any of the following ways: oral, inhalation spray, rectal, nasal, buccal, topical, parenteral e.g. subcutaneous, intravenous, intramuscular, intraperitoneal, thecal Intravenous, intraventricular, intrasternal and intracranial injection or infusion, or with the aid of an explanted reservoir. Among them, oral, intraperitoneal or intravenous administration is preferred.

When administered orally, the compounds of the present invention may be prepared in any orally acceptable preparation form, including but not limited to tablets, capsules, aqueous solutions or suspensions. Wherein, the carrier that tablet uses generally comprises lactose and cornstarch, also can add lubricating agent such as magnesium stearate in addition. Diluents used in capsule formulations generally include lactose and dried cornstarch. Aqueous suspensions are usually prepared by mixing the active ingredient with suitable emulsifying and suspending agents. If desired, some sweetening, flavoring or coloring agents may also be added to the above oral preparation forms.

The compounds of this invention can also be administered in the form of sterile injectable preparations, including sterile injectable aqueous or oily suspensions or sterile injectable solutions. Among them, usable vehicles and solvents include water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils, such as mono- or diglycerides, can be employed as a solvent or suspending medium.

In addition, it should be pointed out that the dosage and method of use of the compounds of the present invention depend on many factors, including the patient's age, body weight, sex, natural health status, nutritional status, activity of the compound used, time of administration, metabolic rate, severity of the disease and the subjective judgment of the treating physician. The preferred dose is 0.01-100 mg/kg body weight/day, and the most preferred dose is 5-10 mg/kg body weight/day.

Detailed ways

The present invention is further illustrated by the following examples, which are not intended to limit the invention.

The melting points of the compounds were determined by a YRT-3 melting point apparatus, and the temperature was not corrected. ^{The 1} H-NMR spectrum was measured by a Bruker ARX 400 nuclear magnetic analyzer. The FAB mass spectrum was determined by a Zabspect high-resolution magnetic mass spectrometer.

Intermediate preparation

Intermediate 1 mono-p-methoxybenzyl itaconate

19g (0.169mol) of itaconic anhydride and 50ml of p-methoxybenzyl alcohol were reacted at 55-60°C for 40 hours. After the reaction mixture was cooled to room temperature, 150 ml of diethyl ether and saturated aqueous NaHCO ₃ were added to the reaction mixture. The separated aqueous layer was adjusted to pH 3 with dilute hydrochloric acid, and a large amount of solid precipitated out. After filtration, the crude product was recrystallized from ethyl acetate-n-hexane to obtain 38 g of intermediate 1 (mono-p-methoxybenzyl itaconate) as a white solid, with a yield of 90%. mp: 85.5-87.0°C

MS[M ⁺ ]=251.0m/e; ¹ H-NMR (400MHz, CDCl ₃ )δ7.24(d, 2H, J=8.4Hz), 6.8(d, 2H, J=8.8Hz), 6.43(s , 1H), 5.7(d, 1H, J=0.8Hz), 5.0(s, 2H), 3.7(s, 3H), 3.3(s, 2H)

Intermediate 2 2-Methylene-3-methylsuccinate mono-p-methoxybenzyl ester

To a solution of 2.5 g (10 mmol) of Intermediate 1 in 50 ml of anhydrous THF at -78°C, 10 ml (2M, 20 mmol) of LDA in heptane was added dropwise. After keeping the reaction at -78°C for 1 hour, a solution of 1.42 g of methyl iodide in 10 ml of DMF was added dropwise. After 2 hours, 30ml of 1.5mol/L sulfuric acid aqueous solution was added to the reaction mixture to terminate the reaction, and extracted with ether. The ether layer was washed with saturated NaCl aqueous solution until neutral, and dried over anhydrous magnesium sulfate. After spinning to dryness, the crude product was separated by column to obtain 2.38 g of oily intermediate 2 (2-methylene-3-methylsuccinic acid mono-p-methoxybenzyl ester), with a yield of 90%.

MS[M ⁺ ]=265.0m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ7.25(d, 2H), 6.85(d, 2H), 6.46(s, 1H), 5.80(s, 1H) , 5.07(s, 2H), 3.78(s, 3H), 3.61(q, 1H), 1.39(d, 3H)

General procedure for the preparation of 2-alkyl(or alkenyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl esters from intermediate 2

A solution of 1.43 g (5.42 mmol) of intermediate 2 in 10 ml of anhydrous THF was added dropwise to a solution of LDA 2.71 ml (2M in heptane, 5.42 mmol) in 30 ml of anhydrous THF at -78°C. After stirring the reaction for 2 hours, a solution of 6.5 mmol aldehyde (R ₁ CHO, where R 1 has the above meaning) in anhydrous THF was added dropwise at -78°C. After keeping at -78°C for 6 hours, 20 ml of 1.5 mol/L sulfuric acid aqueous solution was added to the reaction mixture to terminate the reaction. The reaction mixture was extracted with ether, and the ether layer was washed with saturated NaCl aqueous solution until neutral, and dried over anhydrous magnesium sulfate. Spin to dry, and the crude product is separated by column to obtain trans and cis 2-alkyl (or alkenyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxy benzyl ester.

trans-intermediate 3 (trans- 2-hexyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester ) and cis-intermediate 3 ( cis- 2-Hexyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester )

trans-intermediate 3 cis-intermediate 3

Using n-heptanal as a raw material, the trans-intermediate 3 and the cis-intermediate 3 were prepared according to the above-mentioned general operation. MS[M ⁺ ]＝361.0m/e

trans-intermediate 4 (trans- 2-heptyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester ) and cis-intermediate 4 (cis- 2-heptyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester )

Trans-Intermediate 4 cis-Intermediate 4

Using n-octanal as the raw material, it was prepared according to the above general operation to obtain trans-intermediate 4 and cis-intermediate 4. MS[M ⁺ ]＝375.0m/e

trans-intermediate 5 (trans- 2-octyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester ) and cis-intermediate 5 (cis- 2-octyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester )

trans-intermediate 5 cis-intermediate 5

Using n-nonanal as raw material, the preparation was carried out according to the above-mentioned general operation to obtain trans-intermediate 5 and cis-intermediate 5. MS[M ⁺ ]＝389.0m/e

trans-intermediate 6 (trans- 2-nonyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester ) and cis-intermediate 6 (cis- 2-Nonyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester )

trans-intermediate 6 cis-intermediate 6

Using n-decyl aldehyde as a raw material, it was prepared according to the above-mentioned general operation to obtain trans-intermediate 6 and cis-intermediate 6. MS[M ⁺ ]＝403.0m/e

trans-intermediate 7 (trans- 2-decyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester ) and cis-intermediate 7 (cis- 2-decyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester )

trans-intermediate 7 cis-intermediate 7

Using n-undecanal as the raw material, the trans-intermediate 7 and the cis-intermediate 7 were prepared according to the above-mentioned general operation. MS[M ⁺ ]＝417.0m/e

trans-intermediate 8 (trans- 2-n-undecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester ) and cis- Intermediate 8 (cis- 2-n-undecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester )

trans-intermediate 8 cis-intermediate 8

Using n-dodecanal as the starting material, the trans-intermediate 8 and the cis-intermediate 8 were prepared according to the above-mentioned general operation. MS[M ⁺ ]＝431.0m/e

trans-intermediate 9 (trans- 2-n-dodecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester ) and cis- Intermediate 9 (cis- 2-n-dodecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester )

trans-intermediate 9 cis-intermediate 9

Using n-tridecanal as the starting material, the trans-intermediate 9 and the cis-intermediate 9 were prepared according to the above-mentioned general operation. MS[M ⁺ ]＝445.0m/e

trans-intermediate 10 (trans- 2-n-tridecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester ) and cis- Intermediate 10 (cis- 2-n-Tridecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester )

Trans-Intermediate 10 Cis-Intermediate 10

The trans-intermediate 10 and the cis-intermediate 10 were obtained by using n-tetradecaldehyde as the raw material and following the above-mentioned general operation. MS[M ⁺ ]＝459.0m/e

trans-Intermediate 11 (trans- 2-(dec-9-enyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester ) and cis-intermediate 11 (cis- 2-(dec-9-enyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester )

Trans-Intermediate 11 Cis-Intermediate 11

Using 10-en-n-undecanal as a starting material, the trans-intermediate 11 and the cis-intermediate 11 were obtained by following the general procedure described above. MS[M ⁺ ]＝415.0m/e

trans-Intermediate 12 (trans- 2-(cis-non-3-enyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester ) and cis-intermediate 12 (cis- 2-(cis-non-3-enyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxy benzyl ester )

trans-intermediate 12 cis-intermediate 12

Using cis-4-ene n-decyl aldehyde as starting material, it was prepared according to the above general operation to obtain trans-intermediate 12 and cis-intermediate 12. MS[M ⁺ ]=401.0m/e

trans-Intermediate 13 (trans- 2-(3-Benzyloxypropyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxybenzyl ester ) and cis-intermediate 13 (cis- 2-(3-benzyloxypropyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid p-methoxy benzyl ester )

trans-intermediate 13 cis-intermediate 13

Using 4-benzyloxy-n-butyraldehyde as starting material, the trans-intermediate 13 and cis-intermediate 13 were prepared according to the above general procedure. MS[M ⁺ ]＝425.0m/e

Example 1: trans-2-n-hexyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Dissolve 0.68g of trans-intermediate 3 in 40ml of dichloromethane, add 2.2ml of M trifluoroacetic acid aqueous solution, and stir at room temperature overnight. Saturated NaHCO ₃ aqueous solution was added to the reaction mixture, the separated aqueous layer was acidified and extracted with ether, the ether layer was washed with saturated NaCl aqueous solution until neutral, and dried over anhydrous sodium sulfate. The crude product was separated by silica gel column (n-hexane/ethyl acetate/glacial acetic acid) to obtain 0.40 g trans-2-n-hexyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran- 3-Carboxylic acid, yield 88.2%.

MS[M ⁺ ]=241.0m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ10.38(brs, 1H,), 6.39(s, 1H), 5.90(s, 1H), 4.84(dd, 1H ), 1.62～1.26(m, 13H), 0.89(t, 3H)

Example 2: trans-2-n-heptyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, change the trans-intermediate 3 to trans-intermediate 4 to obtain oily trans-2-n-heptyl-3-methyl-4-methylene-5 -Oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=255.0m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ9.71(brs, 1H), 6.38(s, 1H), 5.90(s, 1H), 4.83(dd, 1H) , 1.62～1.28(m, 15H), 0.88(t, 3H)

Example 3: trans-2-n-octyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, change the trans-intermediate 3 to trans-intermediate 5 to obtain oily trans-2-n-octyl-3-methyl-4-methylene-5 -Oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=269.1m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.38(s, 1H), 5.90(s, 1H), 4.82(dd, 1H), 1.62～1.27(m, 17H), 0.88(t, 3H)

Example 4: trans-2-n-nonyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, change the trans-intermediate 3 to trans-intermediate 6 to obtain trans-2-n-nonyl-3-methyl-4-methylene as light yellow solid -5-Oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=283.1m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.38(s, 1H), 5.89(s, 1H), 4.82(dd, 1H), 1.62～1.27(m, 19H), 0.88(t, 3H)

Example 5: trans-2-n-decyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, change the trans-intermediate 3 to trans-intermediate 7 to obtain trans-2-n-decyl-3-methyl-4-methylene as light yellow solid -5-Oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=297.1m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.38(s, 1H), 5.89(s, 1H), 4.83(dd, 1H), 1.62～1.26(m, 21H), 0.88(t, 3H)

Example 6: trans-2-n-undecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, the trans-intermediate 3 was changed to trans-intermediate 8 to obtain trans-2-n-undecyl-3-methyl-4-ylidene as light yellow solid methyl-5-oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=311.0m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.38(s, 1H), 5.89(s, 1H), 4.82(dd, 1H), 1.62～1.26(m, 23H), 0.88(t, 3H)

Example 7: trans-2-n-dodecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, the trans-intermediate 3 was changed to trans-intermediate 9 to obtain trans-2-n-dodecyl-3-methyl-4-ylidene methyl-5-oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=325.1m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.38(s, 1H), 5.89(s, 1H), 4.82(dd, 1H), 1.62～1.26(m, 25H), 0.88(t, 3H)

Example 8: trans-2-n-tridecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, the trans-intermediate 3 was changed to trans-intermediate 10 to obtain trans-2-n-tridecyl-3-methyl-4-ylidene as light yellow solid methyl-5-oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=339.0m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.38(s, 1H), 5.89(s, 1H), 4.82(dd, 1H), 1.62～1.22(m, 27H), 0.88(t, 3H)

Example 9: trans-2-(dec-9-enyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, the trans-intermediate 3 was changed to trans-intermediate 11 to obtain oily trans-2-(dec-9-enyl)-3-methyl-4- Methylene-5-oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=295.0m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.39(s, 1H), 5.91(s, 1H), 5.82(m, 1H), 5.03～4.92(m, 2H), 4.83(dd, 1H), 2.05(q, 2H), 1.64～1.26(m, 17H)

Example 10: trans-2-(cis-non-3-enyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, the trans-intermediate 3 was changed to trans-intermediate 12 to obtain oily trans-2-(cis-non-3-enyl)-3-methyl- 4-Methylene-5-oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=281.0m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.40(s, 1H), 5.91(s, 1H), 5.49～5.43(m, 1H), 5.37～5.31( m, 1H), 4.84(dd, 1H), 2.35～2.21(m, 2H), 2.05(q, 2H), 1.65(q, 2H), 1.44(s, 3H), 1.38～1.26(m, 6H) , 0.88(t,3H)

Example 11: trans-2-[3-(Benzyloxy)-propyl]-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, the trans-intermediate 3 was changed to trans-intermediate 13 to obtain trans-2-[3-(benzyloxy)-propyl]- 3-Methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=305.0m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ7.36～7.26(m, 5H), 6.36(s, 1H), 5.87(s, 1H), 4.85(dd, 1H), 4.50(s, 2H), 3.61～3.48(m, 2H), 1.95～1.62(m, 4H), 1.43(s, 3H)

Example 12: cis-2-n-hexyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, change the trans-intermediate 3 to cis-intermediate 3 to obtain oily cis-2-n-hexyl-3-methyl-4-methylene-5- Oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=241.0m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.36(s, 1H), 5.70(s, 1H), 4.14(dd, 1H), 1.71～1.29(m, 13H), 0.89(t, 3H)

Example 13: cis-2-n-heptyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, change the trans-intermediate 3 to cis-intermediate 4 to obtain oily cis-2-n-heptyl-3-methyl-4-methylene-5 -Oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=255.1m/e; ¹ H-NMR (400MHz, CDCl ₃ )δ～9.30(brs, 1H), 6.36(s, 1H), 5.70(s, 1H), 4.14(dd, 1H) , 1.71～1.27(m, 15H), 0.88(t, 3H)

Example 14: cis-2-n-octyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, change the trans-intermediate 3 to cis-intermediate 5 to obtain oily cis-2-n-octyl-3-methyl-4-methylene-5 -Oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=269.1m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.36(s, 1H), 5.70(s, 1H), 4.14(dd, 1H), 1.70～1.27(m, 17H), 0.88(t, 3H)

Example 15: cis-2-n-nonyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, change the trans-intermediate 3 to cis-intermediate 6 to obtain oily cis-2-n-nonyl-3-methyl-4-methylene-5 -Oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]= 283.1m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.36(s, 1H), 5.70(s, 1H), 4.14(dd, 1H), 1.71～1.26(m, 19H), 0.88(t, 3H)

Example 16: cis-2-n-decyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, change the trans-intermediate 3 to cis-intermediate 7 to obtain oily cis-2-n-decyl-3-methyl-4-methylene-5 -Oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=297.1m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.36(s, 1H), 5.70(s, 1H), 4.13(dd, 1H), 1.72～1.26(m, 21H), 0.88(t, 3H)

Example 17: cis-2-n-decyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, change the trans-intermediate 3 to cis-intermediate 8 to obtain cis-2-n-undecyl-3-methyl-4-methylene as a white solid Base-5-oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=311.1m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.36(s, 1H), 5.70(s, 1H), 4.13(dd, 1H), 1.70～1.21(m, 23H), 0.88(t, 3H)

Example 18: cis-2-n-dodecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, change the trans-intermediate 3 to cis-intermediate 9 to obtain cis-2-n-dodecyl-3-methyl-4-methylene as a white solid Base-5-oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=325.2m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.36(s, 1H), 5.70(s, 1H), 4.13(dd, 1H), 1.70～1.26(m, 25H), 0.88(t, 3H)

Example 19: cis-2-n-tridecyl-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, change the trans-intermediate 3 to cis-intermediate 10 to obtain cis-2-n-tridecyl-3-methyl-4-methylene as a white solid Base-5-oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=339.1m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.36(s, 1H), 5.70(s, 1H), 4.13(dd, 1H), 1.72～1.26(m, 27H), 0.88(t, 3H)

Example 20: cis-2-(dec-9-enyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, the trans-intermediate 3 was changed to cis-intermediate 11 to obtain oily cis-2-(dec-9-enyl)-3-methyl-4- Methylene-5-oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=295.2m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.36(s, 1H), 5.81(m, 1H), 5.70(s, 1H), 5.02～4.91(m, 2H), 4.13(dd, 1H), 2.04(q, 2H), 1.72～1.29(m, 17H)

Example 21: cis-2-(cis-non-3-enyl)-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, the trans-intermediate 3 was changed to cis-intermediate 12 to obtain oily cis-2-(cis-non-3-enyl)-3-methyl- 4-Methylene-5-oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=281.1m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ6.36(s, 1H), 5.70(s, 1H), 5.49～5.43(m, 1H), 5.34～5.27( m, 1H), 4.13(dd, 1H), 2.33～2.22(m, 2H), 2.03(q, 2H), 1.81～1.64(m, 2H), 1.48(s, 3H), 1.37～1.24(m, 6H), 0.87(t, 3H)

Example 22: cis-2-[3-(Benzyloxy)-n-propyl]-3-methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid

Using the preparation method of Example 1, change the trans-intermediate 3 to cis-intermediate 13 to obtain oily cis-2-[3-(phenylmethoxy)-n-propyl]-3 -Methyl-4-methylene-5-oxo-tetrahydrofuran-3-carboxylic acid.

MS[M ⁺ ]=305.0m/e; ¹ H-NMR (400MHz, CDCl ₃ ) δ7.36～7.27(m, 5H), 6.34(s, 1H), 5.68(s, 1H), 4.50(q, 2H), 4.14(dd, 1H), 3.58～3.46(m, 2H), 1.94～1.72(m, 4H), 1.47(s, 3H)

Inhibition test of compounds on fatty acid synthase activity:

Preparation of fatty acid synthase (FAS):

Male SD rats, weighing 220-250 grams, were fasted for two days, then given high-sugar and low-fat feed, and fed for three days. The rat model with high fatty acid expression induced by diet was sacrificed quickly, and the liver was harvested, and the liver was lavaged with ice-cold phosphate buffer (0.1M; pH 7.5) (Note: the above steps were all performed at 4°C). After weighing, the liver was homogenized with 0.1 M phosphate buffer at pH 7.2 at a ratio of 5 ml/mg liver. After fully homogenized, 25% (w/v) polyethylene glycol (Mn is 6000) was added to a concentration of 5%, stirred for 10 minutes, and centrifuged at 9000 rpm for 30 minutes. The supernatant was filtered through a large filter, and its volume was measured, 25% PEG was added thereto to a final concentration of 11%, stirred for 10 minutes and then centrifuged for 15 minutes at 9000 rpm. After centrifugation, the supernatant was discarded, and the surface of the pellet was gently washed with ice-cold distilled water to clean the PEG on the surface as much as possible. The pellet was homogenized with 0.1 M phosphate buffered saline (pH 7.2, containing 1 mM DTT, 2% rabbit serum, 2 mg/ml bovine serum albumin), and the volume was 1/4 of the volume before the last centrifugation. After homogenization, add ammonium sulfate (25g/100ml) while stirring, stir for 30 minutes, and centrifuge at 9000rpm for 30 minutes. Homogenize the pellet with 0.05M phosphate buffered saline (pH 7.2, containing 1mMDTT, 2mg/ml bovine serum albumin), and the final volume is the same as in the previous step. Then, add 25% PEG to the homogenate to a final concentration of 10%, stir for 10 minutes, centrifuge at 9000rpm for 15 minutes, collect and save the supernatant, repeat the previous step for the precipitation, and collect the two supernatants together , adding 35% PEG therein to a final concentration of 15%, stirring for 10 minutes, centrifuging at 9000 rpm for 30 minutes, and collecting the precipitate. The precipitate was dissolved in 0.05M phosphate buffered saline (pH 7.5, containing 2mMDTT), centrifuged for 5 minutes at 3000rpm, and the supernatant was collected. The supernatant was separated by an ion exchange column, and the eluent was 0.05M phosphate buffered saline. The collected liquid was centrifuged and concentrated with an ultrafiltration concentrating tube at a speed of 3000 g to a final volume of about 2 ml. Add the concentrated product to a gel chromatography exchange column for further separation and purification. The eluent was 0.1M phosphate buffered saline (pH 7.5, containing 5mMDTT), and the active peak was collected. Concentrate the collected liquid in the same way as above, and store it, which can be frozen or stored at 4°C. All the above steps were carried out at 4°C.

Inhibition of fatty acid synthase (FAS) by compounds:

Incubate the tested compound with NADPH (200nM), acetyl-CoA (66nM), phosphate buffer (580ml, 0.1M, pH 7.0), 24ul of the above purified fatty acid synthase concentrate, at 37°C for 30min , the reaction was initiated by adding propionyl-CoA (200 nM). Using a S500P spectrophotometer, measure the amount of NADPH oxidized at 340nm, thereby calculating the activity change of FAS. C75 was used as a positive compound.

Inhibitory effect of some compounds on FAS activity

compound   FAS residual activity (%) compound   FAS residual activity (%) Compound Concentration (100ug/ml) Compound Concentration (10ug/ml) Compound Concentration (100ug/ml) Compound Concentration (10ug/ml) Blank 100 100 C75 2.7 50.0 Example 1 76.4 91.7 Example 12 67.6 97.9 Example 2 12.2 88.6 Example 13 76.9 102 Example 3 24.8 76.8 Example 14 37.3 87.3 Example 4 5.0 86.1 Example 15 33.8 88.3 Example 5 3.0 88.3 Example 16 3.0 75.0 Example 6 6.9 60.0 Example 17 9.0 75.0 Example 7 5.8 8.0 Example 18 1.0 52.6

Claims (8)

1. the compound of general formula I comprises its raceme or optically active isomer, or its pharmacy acceptable salt or solvate,

Wherein:

R ₁Be C ₃～C ₁₈Straight or branched alkyl, or C ₃～C ₁₈The straight or branched thiazolinyl, optional on the described alkyl or alkenyl chain have the halogen of being selected from, itrile group, a C ₁～C ₆Straight or branched alkoxyl group, C ₂～C ₆Straight or branched alkene oxygen base, trifluoromethoxy, phenoxy group, benzyloxy, C ₃～C ₆Cycloalkyl, 5～6 yuan aromatic carbocyclic or the substituting group of heterocyclic group, described fragrant heterocycle comprise that 1～4 is selected from O, the heteroatoms of S and N; Can not have replacement on aromatic carbocyclic or the heterocycle, can be selected from following substituting group yet and replace: halogen, nitro, hydroxyl, methylol, trifluoromethyl, trifluoromethoxy, C by 1～5 ₁～C ₆The straight or branched alkyl, C ₂～C ₆The straight or branched thiazolinyl, C ₁～C ₄Alkoxyl group, C ₂～C ₄Alkene oxygen base, phenoxy group, benzyloxy, itrile group, carboxyl or amino;

R ₂Be selected from H, C ₁～C ₆The alkyl of straight or branched, trifluoromethyl has the substituent benzyl of the group of being selected from down: halogen, nitro, hydroxyl, methylol, trifluoromethyl, trifluoromethoxy, C on benzyl and the phenyl ring ₁～C ₆The straight or branched alkyl, C ₂～C ₆The straight or branched thiazolinyl, C ₁～C ₄Alkoxyl group, C ₂～C ₄Alkene oxygen base, phenoxy group, benzyloxy, itrile group, carboxyl and amino;

R ₃For=CH ₂,=CZ ₂, or=CHZ, wherein, Z is C ₁～C ₃The straight or branched alkyl, F, Cl, or Br;

X is selected from O and S atom;

Y is selected from halogen, C ₁～C ₄The straight or branched alkyl, C ₂～C ₄The straight or branched thiazolinyl, C ₁～C ₄Alkoxyl group, C ₂～C ₄Alkene oxygen base, itrile group, nitro, hydroxyl, methylol, trifluoromethyl, trifluoromethoxy, and amino.

2. the compound of claim 1, it is the compound of general formula I I representative, comprises its raceme or optically active isomer, or its pharmacy acceptable salt or solvate,

Wherein:

R ₁Be C ₃～C ₁₈Straight or branched alkyl, or C ₃～C ₁₈The straight or branched thiazolinyl, optional on its chain have the halogen of being selected from, itrile group, a C ₁～C ₆Straight or branched alkoxyl group, C ₂～C ₆Straight or branched alkene oxygen base, trifluoromethoxy, phenoxy group, benzyloxy, C ₃～C ₆Cycloalkyl, 5～6 yuan aromatic carbocyclic or the substituting group of heterocyclic group comprise in the described fragrant heterocycle that 1～4 is selected from O, the heteroatoms of S and N; Can not have replacement on aromatic carbocyclic or the heterocycle, can be selected from following substituting group yet and replace: halogen, nitro, hydroxyl, methylol, trifluoromethyl, trifluoromethoxy, C by 1～5 ₁～C ₆The straight or branched alkyl, C ₂～C ₆The straight or branched thiazolinyl, C ₁～C ₄Alkoxyl group, C ₂～C ₄Alkene oxygen base, phenoxy group, benzyloxy, itrile group, carboxyl and amino;

X is selected from O and S atom.

3. claim 1 or 2 compound, it is selected from:

Trans-the 2-n-hexyl-the 3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Trans-the 2-n-heptyl-the 3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Trans-the 2-n-octyl-the 3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Trans-the 2-n-nonyl-the 3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

The positive decyl of trans-2--3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Trans-2-n-undecane base-the 3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Trans-the 2-dodecyl-the 3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Trans-2-n-tridecane base-the 3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Trans-2-(last of the ten Heavenly stems-the 9-thiazolinyl)-3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Trans-2-(suitable-ninth of the ten Heavenly Stems-the 3-thiazolinyl)-3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Trans-2-[3-(benzyloxy)-propyl group]-3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Cis-2-n-hexyl-3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Cis-2-n-heptyl-3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Cis-2-n-octyl-3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Cis-2-n-nonyl-3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

The positive decyl of cis-2--3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Cis-2-n-undecane base-3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Cis-2-dodecyl-3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Cis-2-n-tridecane base-3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Cis-2-(last of the ten Heavenly stems-the 9-thiazolinyl)-3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Cis-2-(suitable-ninth of the ten Heavenly Stems-the 3-thiazolinyl)-3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

Cis-2-[3-(benzyloxy)-n-propyl]-3-methyl-4-methylene radical-5-oxo-tetrahydrofuran (THF)-3-carboxylic acid;

With and pharmacy acceptable salt and solvate.

4. pharmaceutical composition, it contains each described compound of claim 1～3, comprises its raceme or optically active isomer, or its pharmacy acceptable salt or solvate and at least a pharmaceutically acceptable carrier, thinner or vehicle.

5. the method for preparation formula Ia compound, it comprises:

1) make itaconic anhydride (III) and p-methoxybenzyl alcohol (IV) reaction,

Get the compound of formula V,

2) with formula V compound in the presence of alkali (as: lithium diisopropylamine (LDA)) with methyl iodide (CH ₃I) reaction gets formula VI compound.

3) make formula VI compound in the presence of alkali (as: lithium diisopropylamine (LDA)) with aldehyde R ₁CHO reaction, wherein R ₁Definition with claim 1, formula Ia compound and

4) randomly, separation obtains trans respectively and formula Ia compound cis through post,

6. the method for preparation formula IIa compound comprises racemic formula Ia compound hydrolysis is obtained racemic formula IIa compound, perhaps the formula Ia compound hydrolysis of trans or cis is obtained the formula IIa compound of trans or cis respectively,

Wherein, R ₁Definition with claim 1.

7. claim 1～3 each described compound is used to prepare the purposes of slimming medicine.

8. claim 1～3 each described compound is used to prepare the purposes of antitumor drug.