Class of dienyl long-chain compounds acting on ACLY, and preparation method therefor and use thereof

Abstract

Disclosed in the present invention are a class of dienyl long-chain compounds, and a preparation method therefor and the use thereof. The structure is as shown in formula I, and the definition of each substituent in the formula is as described in the description and the claims. The compound of the present invention has an ACLY inhibitory activity and a lipid synthesis inhibitory activity, and can be used in the preparation of a drug for treating metabolic diseases such as hyperlipidemia and atherosclerosis, or a variety of cancers such as lung cancer, pancreatic cancer, breast cancer, ovarian cancer, liver cancer, intestinal cancer, brain cancer and acute myeloid leukemia.

Description

A class of diene-based long-chain compounds that act on ACLY, their preparation methods and applications Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a class of diene-based long-chain compounds that act on ACLY, their preparation methods, and uses. Background Technology

[0002] ATP citrate lyase (ACLY) plays a crucial role in glucose and lipid metabolism. Acetyl-CoA (Ac-CoA), produced in mitochondria, cannot directly cross the mitochondrial membrane into the cytoplasm. However, Ac-CoA enters the tricarboxylic acid cycle (TCA cycle). Citrate produced in the TCA cycle is transported to the cytoplasm via citrate transport proteins on the mitochondrial membrane. In the cytoplasm, citrate and CoA (CoA) are catalyzed by ACLY to produce acetyl-CoA and oxaloacetate, consuming one molecule of ATP. The biological functions of acetyl-CoA in vivo can be summarized in three aspects: In the fatty acid synthesis pathway, it can be carboxylated by acetyl-CoA carboxylase (ACC) to form malonyl-CoA, which is then further processed by related fatty acid synthases to ultimately produce fatty acids; acetyl-CoA is also a precursor in the mevalonate pathway, which synthesizes farnesyl pyrophosphate (FPP), which participates in cholesterol synthesis; additionally, acetyl-CoA provides raw materials for acetylation reactions, participating in the acetylation of various proteins, including histones. ACLY participates in lipid synthesis and epigenetic regulation in the body by regulating acetyl-CoA.

[0003] Hyperlipidemia, atherosclerosis, and non-alcoholic fatty liver disease are all related to increased lipid synthesis levels. ACLY, as the main enzyme that produces Ac-CoA in the cytoplasm, provides raw materials for lipid synthesis and is a key link in lipid synthesis. The drug Bempedoic acid (ETC-1002), which acts on ACLY, can significantly reduce the level of lipid synthesis in the liver.

[0004] Metabolic remodeling is the most common and primary characteristic of cancer cells. This is because the rapid growth of cancer cells requires large amounts of energy and macromolecules, and their metabolism undergoes significant changes to meet this demand. Since cancerous tissue often develops into irregularly shaped masses, it becomes extremely difficult for cancer cells to obtain lipids from surrounding blood vessels. Their own de novo lipid synthesis is usually at a high level, serving as the main source of lipids required for proliferation. Abnormal epigenetic regulation is another major characteristic of cancer cells. As a key link in epigenetic regulation, histone acetylation plays a crucial role. Elevated histone acetylation levels result in loose chromatin structure, indicating a state of transcriptional activation; conversely, low histone acetylation levels result in tight chromatin structure, indicating a state of transcriptional repression. Studies have shown that the occurrence of many cancers is closely related to elevated histone acetylation levels. Acetyl-CoA, catalyzed by ACLY, is both a raw material for lipid synthesis and participates in histone acetylation. Research indicates that ACLY is highly expressed in various cancers, including non-small cell lung cancer, breast cancer, and liver cancer, and high ACLY expression is closely related to poor prognosis in these cancers.

[0005] In conclusion, the development of ACLY inhibitors holds promise for the clinical treatment of metabolic diseases and cancer. Summary of the Invention

[0006] The purpose of this invention is to provide a class of compounds with ACLY inhibitory activity or pharmaceutically acceptable salts thereof. "Pharmaceutically acceptable salts" include, but are not limited to, sodium, potassium, magnesium, and calcium salts formed from the carboxylic acid groups in the compounds, or inorganic acid salts formed when the series of compounds contains nitrogen.

[0007] In a first aspect, a compound of general formula I, or a stereoisomer, enantiomer, diastereomer, racemate, or a pharmaceutically acceptable salt thereof, is provided.

[0008] In the formula,

[0009] Each double bond can be independently configured in either cis (Z) or trans (E) configuration;

[0010] R1 and R2 are each independently H, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C7 cycloalkyl, C3-C7 cycloalkenyl, C6 or C10 aryl; or R1 and R2 together with the connected carbon form a saturated 3-7 membered ring, or a 3-7 membered ring containing an unsaturated double bond;

[0011] R3 and R4 are each independently H, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C7 cycloalkyl, C3-C7 cycloalkenyl, C6 or C10 aryl; or R3 and R4 together with the connected carbon form a saturated 3-7 membered ring, or a 3-7 membered ring containing an unsaturated double bond;

[0012] m, n, and q are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9;

[0013] The aforementioned C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C7 cycloalkyl, C3-C7 cycloalkenyl, or C6-C10 aryl groups are either unsubstituted or substituted, wherein the substitution refers to being substituted by 1, 2, 3, 4, or 5 substituents selected from the group consisting of: halogen, C1-C4 alkyl, C1-C4 alkoxy, C3-C7 cycloalkyl, C3-C7 cycloalkenyl, and C6-C10 aryl.

[0014] In another preferred embodiment, R1 and R2 are each independently H, methyl, ethyl, propyl, vinyl, propenyl, ethynyl, propynyl, C3-C6 cycloalkyl, C3-C6 cycloalkenyl, or C6-C10 aryl; or, R1 and R2 together with the connected carbon form a saturated 3-6 membered ring, or a 3-6 membered ring containing an unsaturated double bond.

[0015] In another preferred embodiment, R3 and R4 are each independently H, methyl, ethyl, propyl, vinyl, propenyl, ethynyl, propynyl, C3-C6 cycloalkyl, C3-C6 cycloalkenyl, or C6-C10 aryl; or, R3 and R4 together with the connected carbon form a saturated 3-6 membered ring, or a 3-6 membered ring containing an unsaturated double bond.

[0016] In another preferred embodiment, R1 and R2 together with the connected carbon form a saturated four-membered ring, a saturated five-membered ring, a saturated six-membered ring, a four-membered ring containing an unsaturated double bond, a five-membered ring containing an unsaturated double bond, or a six-membered ring containing an unsaturated double bond.

[0017] In another preferred embodiment, R3 and R3 together with the connected carbon form a saturated four-membered ring, a saturated five-membered ring, a saturated six-membered ring, a four-membered ring containing an unsaturated double bond, a five-membered ring containing an unsaturated double bond, or a six-membered ring containing an unsaturated double bond.

[0018] In another preferred embodiment, R1 and R2 are H; or R1 and R2 together with the connected carbon form a four-membered ring containing an unsaturated double bond or a five-membered ring containing an unsaturated double bond.

[0019] In another preferred embodiment, R3 and R4 are H; or R1 and R2 together with the connected carbon form a four-membered ring containing an unsaturated double bond or a five-membered ring containing an unsaturated double bond.

[0020] In another preferred embodiment, the compound has the structure shown in Formula II:

[0021] Where m, n and q are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8 or 9.

[0022] In another preferred example, q = 0, 1, 2 or 3.

[0023] In another preferred example, q = 0 or 1.

[0024] In another preferred embodiment, m and n are each independently 4, 5, 6, 7, 8, or 9.

[0025] In another preferred embodiment, m and n are each independently 6, 7, 8 or 9, preferably 7 or 8.

[0026] In another preferred embodiment, the two double bonds are each independently in either a cis configuration (Z) or a trans configuration (E).

[0027] In another preferred embodiment, the compound is selected from the group consisting of:

[0028] In another preferred embodiment, compound M1 has the EE configuration.

[0029] In another preferred embodiment, the M3 compound has a ZE configuration.

[0030] In another preferred embodiment, the M10 compound has a ZE configuration.

[0031] In another preferred embodiment, the M13 compound has a ZZ configuration.

[0032] In a second aspect, the present invention provides a pharmaceutical composition comprising:

[0033] Compounds of formula (I) or formula (II) as described in the first aspect of the invention, or stereoisomers, enantiomers, diastereomers, racemates or pharmaceutically acceptable salts thereof; and pharmaceutically acceptable carriers.

[0034] The dosage form of the drug combination described in this invention can be diverse, including but not limited to: tablets, capsules, granules, syrups, solutions, suspensions or aerosols.

[0035] In a third aspect, the present invention provides the use of a compound of formula (I) or formula (II) as described in the first aspect of the present invention, or a stereoisomer, enantiomer, diastereomer, racemate or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described in the second aspect of the present invention, for the preparation of an ACLY inhibitor; or for the preparation of a medicament for the prevention and / or treatment of metabolic diseases or cancer.

[0036] In another preferred embodiment, the metabolic disease is selected from the group consisting of: hypercholesterolemia, atherosclerosis, non-alcoholic fatty liver disease, and diabetes.

[0037] In another preferred embodiment, the cancer is selected from the group consisting of: lung cancer, pancreatic cancer, breast cancer, ovarian cancer, liver cancer, intestinal cancer, brain cancer, and acute myeloid leukemia.

[0038] In a fourth aspect, the present invention provides a treatment method for using the compound described in the first aspect of the present invention in combination with other anticancer drugs (therapies).

[0039] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Detailed Implementation

[0040] This invention, based on extensive and in-depth research, and through numerous experiments and screenings, unexpectedly discovered for the first time a diene-based long-chain compound. This compound possesses both ACLY inhibitory and lipid synthesis inhibitory activities, demonstrating promising development potential as a therapeutic agent for cancer or metabolic diseases. This invention was completed based on this discovery.

[0041] the term

[0042] To facilitate understanding of the invention, certain technical and scientific terms are specifically defined below. Unless otherwise expressly defined herein, all other technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this invention pertains. Before describing the invention, it should be understood that the invention is not limited to the specific methods and experimental conditions described, as such methods and conditions can vary. It should also be understood that the terminology used herein is intended only to describe particular embodiments and is not intended to be restrictive; the scope of the invention will be limited only by the appended claims.

[0043] As used herein, the term “comprising” or its variations such as “including” or “comprising” are understood to include the said element or component without excluding other elements or other components.

[0044] In this invention, unless otherwise specified, the terms used have the general meanings known to those skilled in the art.

[0045] In this invention, the term "C1-C4" refers to having 1, 2, 3, or 4 carbon atoms. The term "C3-C7" refers to having 3, 4, 5, 6, or 7 carbon atoms, and so on.

[0046] In this invention, the term "alkyl" refers to a saturated linear or branched hydrocarbon moiety. For example, the term "C1-C4 alkyl" refers to a straight-chain or branched alkyl group having 1 to 4 carbon atoms, and includes, without limitation, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.

[0047] In this invention, the term "alkenyl" refers to a straight-chain or branched hydrocarbon moiety containing at least one double bond. For example, the term "C2-C4 alkenyl" refers to a straight-chain or branched alkenyl moiety containing one double bond and having 2 to 4 carbon atoms, and includes, without limitation, vinyl, propenyl, butenyl, and isobutenyl.

[0048] In this invention, the term "alkynyl" refers to a straight-chain or branched alkynyl group containing a triple bond, and includes, without limitation, ethynyl, propynyl, butynyl, isobutynyl, etc.

[0049] In this invention, the term "cycloalkyl" refers to a saturated cyclic hydrocarbon moiety, for example, the term "C3-C7 cycloalkyl" refers to a cyclic alkyl group having 3 to 7 carbon atoms on the ring, and includes, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0050] In this invention, the term "cycloalkenyl" refers to a cyclic hydrocarbon moiety containing at least one double bond. For example, the term "C3-C7 cycloalkenyl" refers to a cyclic alkyl group having 3 to 7 carbon atoms on the ring, and includes, without limitation, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, etc.

[0051] In this invention, the term "aryl" refers to a hydrocarbon moiety comprising one or more aromatic rings. For example, the term "C6-C10 aryl" refers to an aromatic cyclic group having 6 to 10 carbon atoms without heteroatoms on the ring, such as phenyl or naphthyl.

[0052] The pharmaceutically acceptable salts described in this invention can be salts formed by anion and a positively charged group on a compound of Formula I. Suitable anions include chloride, bromide, iodide, sulfate, nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate, acetate, malate, toluenesulfonate, tartrate, fumarate, glutamate, glucuronate, lactate, glutarate, or maleate. Similarly, salts can be formed by cations and negatively charged groups on a compound of Formula I. Suitable cations include sodium, potassium, magnesium, calcium, and ammonium ions, such as tetramethylammonium ions.

[0053] The pharmaceutically acceptable carriers described in this invention refer to one or more compatible solid or liquid fillers or gel substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here refers to the ability of the components in the composition to interact with and be mixed with the compounds of this invention without significantly reducing the efficacy of the active ingredient. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as microcrystalline cellulose, cellulose acetate, sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), lactose and its complexes, starch and its derivatives or complexes (such as pregelatinized starch, thermoplastic starch, etc.), gelatin, and pharmaceutical carriers (such as cyclodextrin and its derivatives or complexes, sucrose cores, silica cores, microcrystalline cellulose cores, magnesium aluminum silicate, silica and its complexes). Fillers such as calcium salts (e.g., calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.), oily solvents (e.g., soybean oil, sesame oil, peanut oil, olive oil, medium-chain triglycerides, oleyl alcohol, triacetin, cetearyl alcohol, etc.), polyols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), and emulsifiers (e.g., calcium salts, calcium hydrogen phosphate, calcium carbonate, etc.). Carbomer, Polyethylene glycol, diethylene glycol monoethyl ether, polycetol, etc.), wetting agents (such as sodium dodecyl sulfate, poloxamer, polyoxyethylene castor oil and its derivatives or complexes), Vitamin E (polyethylene glycol succinate, glyceryl monostearate and glyceryl distearate, etc.), disintegrants (such as crospovidone, crospovidone sodium carboxymethyl cellulose, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, etc.), binders (such as povidone, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, etc.), triethyl citrate, talc, film coating materials (such as... Copovidone, etc.), enteric coating materials (such as polyacrylates and their derivatives or complexes, Hydroxypropyl methylcellulose phthalate, cellulose acetate phthalate, etc.), sustained-release coating materials (such as ethyl cellulose and its derivatives or complexes, Solid lubricants (such as stearic acid and its derivatives, magnesium stearate, glyceryl behenate, etc.), pH adjusters, colorants (such as titanium dioxide), flavoring agents (such as aspartame, sucralose, fructose syrup, etc.), stabilizers (such as disodium edetate, gum arabic, tragacanth gum, peach gum, sodium alginate, agar, starch paste, dextran, etc.), antioxidants (such as butylated hydroxytoluene, butylated hydroxyanisole, α-tocopherol, propyl gallate, etc.), preservatives (such as phenoxyethanol, parabens, etc.), pyrogen-free water, etc.

[0054] Preparation method

[0055] The compounds having general formula I described in this invention can be prepared and synthesized via the following route.

[0056] Route 1

[0057] (1) 1-Alynyl alcohol compound A1 reacts with excess 3,4-dihydro-2H-pyran (DHP) under the catalysis of p-toluenesulfonic acid (TsOH) to generate intermediate A3; 1-Alynyl alcohol compound A2 reacts with excess 3,4-dihydro-2H-pyran (DHP) under the catalysis of p-toluenesulfonic acid (TsOH) to generate intermediate A4.

[0058] (2) A3 reacts with iodine (I2) under the conditions of zirconium dichlorodichlorocerocene (Cp2ZrCl2) and diisobutylaluminum hydride (DIBAL-H) ​​to form intermediate A5; A4 reacts with pinacolborane (HBpin) under the conditions of zirconium hydrochloride (Cp2ZrHCl) to form intermediate A6.

[0059] (3) A5 and A6 were catalyzed by chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (XPhos Pd G2) and cesium carbonate (Cs2CO3) to generate intermediate A7.

[0060] (4) Intermediate A7 is deprotected in methanol under the catalysis of TsOH to generate intermediate A8.

[0061] (5) Intermediate A8 undergoes an iodination reaction with iodine, imidazole and triphenylphosphine (PPh3) to generate intermediate A9.

[0062] (6) A9 reacts with an ester containing an active hydrogen at the α-position to form A10 under lithium diisopropylamino (LDA) conditions.

[0063] (7) A10 undergoes hydrolysis in the presence of potassium hydroxide and is further acidified to obtain A11.

[0064] Route 2

[0065] (1) Intermediate A3 and iodine (I2) react under the conditions of indium trichloride (InCl3), diisobutylaluminum hydride (DIBAL-H) ​​and triethylboron (BEt3) to form intermediate A12.

[0066] (2) Intermediate A12 and intermediate A6 were catalyzed by chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (XPhos Pd G2) and cesium carbonate (Cs2CO3) to generate intermediate A13.

[0067] (3) Intermediate A13 is deprotected in methanol under TsOH catalysis to generate intermediate A14.

[0068] (4) Intermediate A14 undergoes an iodination reaction with iodine (I2), imidazole and triphenylphosphine (PPh3) to generate intermediate A15.

[0069] (5) Intermediate A15 reacts with an ester containing an active hydrogen at the α-position under diisopropylaminolithium conditions to generate A16.

[0070] (6) A16 undergoes hydrolysis in the presence of potassium hydroxide and is further acidified to obtain A17.

[0071] Route 3

[0072] (1) Intermediate A4 and intermediate A12 were catalyzed by bis(triphenylphosphine) palladium dichloride (Pd(PPh3)2Cl2) and cuprous iodide (CuI) to generate intermediate A18.

[0073] (2) A18 is converted to A19 by nickel acetate, sodium borohydride and ethylenediamine under hydrogen conditions.

[0074] (3) Intermediate A19 is deprotected in methanol under TsOH catalysis to generate intermediate A20.

[0075] (4) Intermediate A20 undergoes an iodination reaction with iodine (I2), imidazole and triphenylphosphine (PPh3) to generate intermediate A21.

[0076] (5) Intermediate A21 reacts with an ester containing an active hydrogen at the α-position under diisopropylaminolithium conditions to generate A22.

[0077] (6) A22 undergoes hydrolysis in the presence of potassium hydroxide and is further acidified to obtain A23.

[0078] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0079] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0080] instrument

[0081] NMR was performed using a Bruker 400M instrument. NMR calibration: δH 7.26ppm (CDCl3), 2.50ppm (DMSO-d6), 2.05ppm (Acetone-d6). Mass spectrometry was performed using an Agilent 1200 Quadrupole LC / MS system. The silica gel plates for TLC were manufactured by Shandong Yantai Huiyou Silica Gel Development Co., Ltd., model HSGF 254. The normal-phase column silica gel used for compound purification was manufactured by the Qingdao Marine Chemical Plant Branch of Shandong, model ZCX-11, 200-300 mesh. Other commonly used commercial reagents were provided by Shanghai Reagent Company.

[0082] Example 1: Synthesis of M1

[0083] Preparation of compounds B3 and B4

[0084] Under nitrogen protection, 10.00 g of compound B1 (101.89 mmol) was dissolved in 250 mL of dichloromethane (DCM), and 1.94 g of p-toluenesulfonic acid TsOH (10.19 mmol) was added. The mixture was cooled in an ice-water bath, and 12.86 g of 3,4-dihydro-2H-pyran (152.84 mmol) was added dropwise. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until complete. The mixture was diluted with 100 mL of DCM, washed twice with water, once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 50:1) to give 14.09 g of product B3, a colorless oil, with a yield of 75.87%.

[0085] Under nitrogen protection, 10.00 g of compound B2 (118.88 mmol) was dissolved in 250 mL of dichloromethane, and 2.26 g of p-toluenesulfonic acid TsOH (11.89 mmol) was added. The mixture was cooled in an ice-water bath, and 15.00 g of 3,4-dihydro-2H-pyran (178.32 mmol) was added dropwise. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until complete. The mixture was diluted with 100 mL of DCM, washed twice with water, once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 50:1) to give 15.00 g of product B4, a colorless oil, in 75.00% yield.

[0086] (2) Preparation of compound B5

[0087] 943.84 mg of zirconium dichlorocerocene (3.23 mmol) was dissolved in 7.5 mL of dry tetrahydrofuran (THF). Under nitrogen protection, the atmosphere was purged three times. 3.23 mL of diisobutylaluminum hydride (DIBAL-H, 1 M hexane solution) was slowly added dropwise at 0 °C. The reaction was carried out at 0 °C for 10 min, then at room temperature for 30 min. Then, 1.5 mL of dry tetrahydrofuran (THF) solution containing 550.00 mg of compound B3 (3.02 mmol) was added at 0 °C. After 5 min, the reaction was carried out at room temperature for 2 h. 972.69 mg of iodine (3.83 mmol) was dissolved in 4.5 mL of dry tetrahydrofuran (THF) and added dropwise to the above reaction flask at -78 °C. The reaction was carried out at -78 °C for 1 h. The reaction was monitored by TLC until it was complete. The reaction was quenched by adding 10 ml of 1N HCl. The mixture was extracted three times with diethyl ether, and the organic phases were combined. The mixture was washed once with saturated sodium thiosulfate solution and once with saturated brine solution. The mixture was dried over anhydrous sodium sulfate, concentrated, and passed through a column chromatography at a ratio of PE:EA = 50:1 to obtain product B5428.00 mg, a colorless oily substance with a yield of 45.73%.

[0088] (3) Preparation of compound B6

[0089] 400.00 mg of compound B4 (2.38 mmol), 319.50 mg of pinacolborane (2.50 mmol), 61.31 mg of zirconium chlorohydrin (0.24 mmol), and 24.06 mg of triethylamine (0.24 mmol) were placed in a 25 mL round-bottom flask and reacted at 60 °C for 16 h. The reaction was monitored by TLC until complete. The product was diluted with petroleum ether, filtered through diatomaceous earth, concentrated, and column-sected at a PE:EA ratio of 20:1 to give product B6 347.00 mg, a colorless oil, with a yield of 49.27%.

[0090] (4) Preparation of compound B7

[0091] 381.50 mg B5 (1.23 mmol) and 347.00 mg B6 (1.17 mmol) were dissolved in 25 mL of dimethyl sulfoxide solution. The mixture was kept under argon protection with three purging cycles. 92.00 mg Xphos Pd G2 (0.12 mmol) and 2.29 g cesium carbonate (7.03 mmol) were added. The reaction was carried out at room temperature for 20 min, then at 50 °C for 2 h. The reaction was monitored by TLC until complete. 10 mL of saturated ammonium chloride solution was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 50:1) to give 197.00 mg of product B7, a colorless oil, with a yield of 47.71%.

[0092] (5) Preparation of compound B8

[0093] 197.00 mg of compound B7 (0.56 mmol) was dissolved in 10 mL of methanol, and 21.30 mg (0.11 mmol) of p-toluenesulfonic acid (TsOH) was added. The reaction was carried out at room temperature for 4 hours. The reaction was monitored by TLC until complete. The methanol was removed by rotary evaporation, and 20 mL of ethyl acetate was added. The mixture was washed three times with water and once with saturated brine. The solution was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM:MeOH = 50:1) to give product B8 98.50 mg, a colorless oil with a yield of 95.62%.

[0094] (6) Preparation of compound B9

[0095] 350.40 mg of triphenylphosphine (1.34 mmol) and 91.00 mg of imidazole (1.34 mmol) were dissolved in 10 mL of dry dichloromethane. Under nitrogen protection, the mixture was purged and cooled to 0 °C. 339.10 mg of iodine (1.34 mmol) was added under light-protected conditions, and the reaction was carried out at 0 °C for 30 min. 98.50 mg of compound B8 (0.53 mmol) was dissolved in 5 mL of dry dichloromethane and added to the above reaction flask. The mixture was then heated to room temperature and reacted for 4 h. The reaction was monitored by TLC until complete. Most of the dichloromethane was removed by rotary evaporation. 30 mL of diethyl ether was added, and the mixture was washed three times with saturated sodium thiosulfate solution, once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 50:1) to give 176.30 mg of product B9, a colorless oil, with a yield of 81.65%.

[0096] (7) Preparation of compound B11

[0097] 176.30 mg B9 (0.44 mmol) and 304.10 mg B10 (2.62 mmol) were dissolved in 20 mL of dry tetrahydrofuran under nitrogen protection, ventilated, and cooled to -78 °C. 1.31 mL of lithium diisopropylaminolithium LDA (2.0 M 2.62 mmol) was added dropwise, and the solution changed from colorless to yellow. The reaction was allowed to proceed overnight from -78 °C to room temperature. The reaction was monitored by TLC until complete. 40 mL of saturated ammonium chloride solution was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 20:1) to give 89.60 mg of product B11, a colorless oil, with a yield of 53.96%.

[0098] (8) Preparation of compound M1

[0099] 89.60 mg B11 (0.24 mmol) was dissolved in 5 mL of ethanol, and 180.00 mg potassium hydroxide was dissolved in 1 mL of water. These solutions were added to the reaction flask, and the mixture was refluxed overnight. The reaction was monitored by TLC until complete. The ethanol was removed by rotary evaporation, and the mixture was diluted with 10 mL of water. The pH was adjusted to approximately 2 with 1 M hydrochloric acid, resulting in the precipitation of a solid. The solid was extracted three times with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM:MeOH = 50:1) to obtain 70.00 mg of product M1 as a pale yellow solid, with a yield of 91.66%. 1 H NMR (400MHz, CDCl3) δ6.04–5.91(m,2H),5.62(dt,J=14.0,7.1Hz,1H),5.49(dt,J=13.9,7.0Hz,1 H),2.11–1.96(m,4H),1.60–1.48(m,4H),1.43–1.32(m,4H),1.25(p,J=8.2Hz,2H),1.18(s,12H).

[0100] Compound M1 has the EE configuration.

[0101] By replacing different substrates and using a synthetic route similar to M1, the following compounds were obtained.

[0102] Compound M2: 1 H NMR (400MHz, CDCl3) δ5.97(p,J=10.7Hz,2H),5.50(dt,J=14.2,7.3Hz,2H),2.16–2.00(m,4H),1.68(t,J =7.7Hz,2H),1.52(d,J=6.4Hz,2H),1.39(t,J=6.6Hz,2H),1.26(t,J=4.3Hz,4H),1.18(d,J=9.7Hz,12H)

[0103] Example 2 Synthesis of M3

[0104] (1) Preparation of compound B12

[0105] 2.61 g of indium trichloride (11.79 mmol) was dissolved in 50 mL of dry tetrahydrofuran under argon protection with three gas purgings. The mixture was cooled to -78 °C, and 7.53 mL of diisobutylaluminum hydride (1.5 M, 11.29 mmol) was slowly added dropwise. The mixture was reacted at -78 °C for 30 min, then 1.50 g of compound B12 (8.24 mmol) and 1.73 mL of triethylboron (1.0 M, 1.73 mmol) were added. The mixture was reacted at -78 °C for 3 h, and then 13.30 g of iodine (52.42 mmol) was added all at once, and the mixture was reacted for 1 h. The reaction was monitored by TLC until complete. The reaction solution was poured into 100 ml of saturated sodium bicarbonate solution and then into 100 ml of saturated sodium thiosulfate solution. The solution changed from brownish-black to white suspension. The suspension was filtered through diatomaceous earth, extracted three times with ether, washed once with water, washed once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography at a PE:EA ratio of 50:1 to obtain 1.26 g of product B12, a colorless oily substance, with a yield of 49.23%.

[0106] (2) Preparation of compound B13

[0107] 450.00 mg B6 (1.52 mmol) and 494.80 mg B12 (1.60 mmol) were dissolved in 30 mL of dimethyl sulfoxide solution. The mixture was purged three times under argon protection. 119.40 mg Xphos Pd G2 (0.15 mmol) and 2.97 g cesium carbonate (9.12 mmol) were added. The reaction was carried out at room temperature for 20 min and then at 50 °C for 1.5 h. The reaction was monitored by TLC until complete. 20 mL of saturated ammonium chloride solution was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 50:1) to give 255.00 mg of product B13, a colorless oil, with a yield of 47.62%.

[0108] (3) Preparation of compound B14

[0109] 255.00 mg of compound B13 (0.72 mmol) was dissolved in 15 mL of methanol, and 27.50 mg (0.15 mmol) of p-toluenesulfonic acid (TsOH) was added. The reaction was carried out at room temperature for 4 hours. The reaction was monitored by TLC until complete. The methanol was removed by rotary evaporation, and 20 mL of ethyl acetate was added. The mixture was washed three times with water and once with saturated brine. The solution was dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography at a DCM:MeOH ratio of 50:1 to give 128.30 mg of product B14, a colorless oil, with a yield of 96.25%.

[0110] (4) Preparation of compound B15

[0111] 456.50 mg of triphenylphosphine (1.74 mmol) and 118.50 mg of imidazole (1.74 mmol) were dissolved in 15 mL of dry dichloromethane. Under nitrogen protection, the mixture was purged and cooled to 0 °C. 441.80 mg of iodine (1.74 mmol) was added under light-protected conditions, and the reaction was continued at 0 °C for 30 min. 128.30 mg of compound B14 (0.70 mmol) was dissolved in 5 mL of dry dichloromethane and added to the above reaction flask. The mixture was then allowed to react at room temperature for 4 h. The reaction was monitored by TLC until complete. Most of the dichloromethane was removed by rotary evaporation. 30 mL of diethyl ether was added, and the mixture was washed three times with saturated sodium thiosulfate solution, once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 50:1) to give product B15 215.10 mg, a colorless oil, with a yield of 76.46%.

[0112] (5) Preparation of compound B16

[0113] 215.10 mg B16 (0.53 mmol) and 371.00 mg B10 (3.19 mmol) were dissolved in 20 mL of dry tetrahydrofuran under nitrogen protection, ventilated, and cooled to -78 °C. 1.60 mL of lithium diisopropylaminol LDA (2.0 M 3.19 mmol) was added dropwise, and the solution changed from colorless to yellow. The reaction was allowed to proceed overnight from -78 °C to room temperature. The reaction was monitored by TLC until complete. 40 mL of saturated ammonium chloride solution was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 20:1) to give 95.70 mg of product B11, a colorless oil, with a yield of 47.24%.

[0114] (6) Preparation of compound M3

[0115] 95.70 mg B11 (0.25 mmol) was dissolved in 5 mL of ethanol, and 180 mg potassium hydroxide was dissolved in 1 mL of water. These solutions were added to the reaction flask, and the mixture was refluxed overnight. The reaction was monitored by TLC until complete. The ethanol was removed by rotary evaporation, and the mixture was diluted with 10 mL of water. The pH was adjusted to approximately 2 with 1 M hydrochloric acid, resulting in the precipitation of a solid. The solid was extracted three times with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM:MeOH = 50:1) to obtain product M3 78.10 mg as a pale yellow solid, with a yield of 95.72%. 1HNMR (400MHz, CDCl3) δ6.29(dd,J=15.1,11.0Hz,1H),5.95(dd,J=10.9,10.9Hz,1H),5.67(dt,J=14.4,6.8Hz,1H),5.30(dt,J=11.1,8.0Hz,1H),2.16(t d,J=7.4,7.3Hz,2H),2.08(td,J=7.3,7.2Hz,2H),1.60–1.47(m,4H),1.38( dp,J=14.0,7.2Hz,4H),1.29(td,J=9.0,4.6Hz,1H),1.19(d,J=4.6Hz,12H).

[0116] Compound M3 has a ZE configuration.

[0117] By replacing different substrates and using a synthetic route similar to that of M3, the following compounds were obtained.

[0118] Compound M4: 1 H NMR (500MHz, CDCl3) δ6.27(dd,J=15.3,11.0Hz,1H),5.92(dd,J=11.0,11.0Hz,1H),5.60(dt,J=14.6,7.0Hz,1H),5.28(dt,J=10.8, 7.6Hz,1H),2.19–2.09(m,4H),1.69–1.62(m,2H),1.57–1.49(m,2H),1.43–1.35(m,2H),1.34–1.24(m,4H),1.20(d,J=10.3Hz,12H).

[0119] Compound M5: 1 H NMR (600MHz, CDCl3) δ6.22(dd,J=15.3,11.0Hz,1H),5.94(dd,J=11.0,11.0Hz,1H),5.60(dt,J=14.5,6.9Hz,1H),5.29(dt,J=10.7,7. 6Hz,1H),2.14–2.08(m,4H),1.64–1.58(m,2H),1.55–1.49(m,2H),1.45(t,J=6.7Hz,2H),1.32–1.25(m,4H),1.21(d,J=15.8Hz,12H).

[0120] Compound M6: 11H NMR (400 MHz, CDCl3) δ 6.25 (dd, J = 15.0, 10.9 Hz, 1H), 5.96 (dd, J = 10.8, 10.8 Hz, 1H), 5.71 (dt, J = 15.2, 6.1 Hz, 1H), 5.26 (dt, J = 10.8, 7.9 Hz, 1H), 2.18 (q, J = 7.3 Hz, 2H), 2.05 (q, J = 6.7 Hz, 2H), 1.56–1.48 (m, 4H), 1.44–1.26 (m, 6H), 1.23 (q, J = 4.5 Hz, 6H), 1.18 (d, J = 4.9 Hz, 12H).

[0121] Compound M7: 1 1H NMR (400 MHz, CDCl3) δ 6.26 (dd, J = 15.1, 10.9 Hz, 1H), 5.94 (dd, J = 10.9, 10.9 Hz, 1H), 5.60 (td, J = 14.8, 7.3 Hz, 1H), 5.31 (td, J = 13.1, 5.4 Hz, 1H), 2.12 (q, J = 3.3 Hz, 4H), 1.52 (dt, J = 11.8, 3.9 Hz, 4H), 1.44–1.27 (m, 9H), 1.30–1.23 (m, 3H), 1.18 (d, J = 2.1 Hz, 12H).

[0122] Compound M8: 1 1H NMR (400 MHz, CDCl3) δ 6.27 (dd, J = 15.3, 10.9 Hz, 1H), 5.97 (dd, J = 10.8, 10.8 Hz, 1H), 5.81 (dq, J = 4.4, 2.3 Hz, 2H), 5.78–5.67 (m, 3H), 5.26 (td, J = 10.5 Hz, 1H), 2.46–2.32 (m, 6H), 2.19 (q, J = 7.2 Hz, 2H), 2.06 (q, J = 6.9 Hz, 2H), 1.84–1.68 (m, 4H), 1.65–1.48 (m, 2H), 1.44–1.16 (m, 12H).

[0123] Compound M9: 11H NMR (400 MHz, CDCl3) δ 6.27 (dd, J = 15.0, 11.0 Hz, 1H), 5.95 (dd, J = 10.9, 10.9 Hz, 1H), 5.82 (tq, J = 4.2, 2.0 Hz, 2H), 5.72 (tt, J = 5.6, 2.5 Hz, 2H), 5.58 (dt, J = 14.7, 7.1 Hz, 1H), 5.31 (dt, J = 10.8, 7.9 Hz, 1H), 2.39 (q, J = 7.0 Hz, 6H), 2.12 (dq, J = 10.4, 6.2 Hz, 4H), 1.76 (dt, J = 14.4, 6.5 Hz, 4H), 1.65–1.50 (m, 2H), 1.45–1.23 (m, 12H).

[0124] Compound M10: 1 1H NMR (400 MHz, CDCl3) δ 6.20 (dd, J = 15.0, 11.1 Hz, 1H), 5.94 (dd, J = 10.8, 10.8 Hz, 1H), 5.60 (dt, J = 14.6, 7.0 Hz, 1H), 5.35 (dt, J = 11.1, 7.9 Hz, 1H), 2.10 (dd, J = 11.4, 5.3 Hz, 4H), 1.62–1.50 (m, 4H), 1.40–1.29 (m, 4H), 1.29–1.21 (m, 2H), 1.18 (s, 12H).

[0125] Compound M11: 1 1H NMR (400 MHz, CDCl3) δ 6.29 (dd, J = 15.3, 10.9 Hz, 1H), 5.96 (dd, J = 10.9, 10.9 Hz, 1H), 5.65 (dt, J = 14.5, 6.9 Hz, 1H), 5.29 (dt, J = 10.7, 7.7 Hz, 1H), 2.20–1.99 (m, 4H), 1.52 (t, J = 7.5 Hz, 4H), 1.37 (q, J = 7.0 Hz, 4H), 1.27 (s, 20H), 1.18 (s, 12H).

[0126] Compound M12: 1H NMR (400MHz, CDCl3) δ6.29 (dd, J=15.1, 11.0Hz, 1H), 5.96 (dd, J=10.9, 10.9Hz, 1H), 5.81(td,J=5.4,2.2Hz,2H),5.72(td,J=5.9,2.5Hz,2H),5.65(dt,J=14.4,6.8Hz,1H ),5.29(dt,J=10.8,7.9Hz,1H),2.45–2.32(m,6H),2.26–1.98(m,4H),1.76(tt,J=11 .6,6.0Hz,4H),1.59(d,J=6.8Hz,1H),1.38(q,J=7.2Hz,3H),1.26(t,J=3.6Hz,22H).

[0127] Compound M10 has a ZE configuration.

[0128] Example 3 Synthesis of M13

[0129] (1) Preparation of compound B17

[0130] 208.60 mg of bis(triphenylphosphine)palladium dichloride (0.30 mmol) and 169.80 mg of cuprous iodide (0.89 mmol) were dissolved in 1 mL of triethylamine. The mixture was purged three times under nitrogen protection. 500.00 mg of compound B4 (2.97 mmol) and 1.01 g of compound B12 (3.27 mmol) were dissolved in 1 mL of triethylamine and added to the above reaction flask. The reaction was carried out at room temperature for 5 h. The reaction was monitored by TLC until complete. The mixture was diluted with petroleum ether, filtered through diatomaceous earth, concentrated, and column-sected at a PE:EA ratio of 20:1 to give product B17 824.00 mg, a yellow oily substance, with a yield of 79.10%.

[0131] (2) Preparation of compound B18

[0132] 585.00 mg of nickel acetate (2.35 mmol) was suspended in 10 mL of ethanol under argon protection. 88.90 mg of sodium borohydride (2.35 mmol) was added, and the solution turned from pale green to black. The hydrogen balloon was replaced, and the gas was changed several times. 0.24 mL of ethylenediamine was added, followed by dropwise addition of an ethanol solution of 824.00 mg of compound B17. The reaction was carried out at room temperature for 2 hours. The reaction was monitored by TLC until complete. The solution was diluted with 40 mL of ethyl acetate, filtered through diatomaceous earth, and the filtrate was concentrated. The solution was then passed through a column chromatography at a PE:EA ratio of 50:1 to obtain 400.00 mg of product B18, a colorless oil with a yield of 48.27%.

[0133] (3) Preparation of compound B19

[0134] 400.00 mg of compound B18 (1.13 mmol) was dissolved in 20 mL of methanol, and 43.20 mg (0.23 mmol) of p-toluenesulfonic acid (TsOH) was added. The reaction was carried out at room temperature for 4 hours. The reaction was monitored by TLC until complete. The methanol was removed by rotary evaporation, and 30 mL of ethyl acetate was added. The mixture was washed three times with water and once with saturated brine. The solution was dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography at a DCM:MeOH ratio of 50:1 to give product B19 197.40 mg, a colorless oil, with a yield of 94.40%.

[0135] (4) Preparation of compound B20

[0136] 702.40 mg of triphenylphosphine (2.68 mmol) and 182.30 mg of imidazole (2.68 mmol) were dissolved in 25 mL of dry dichloromethane. Under nitrogen protection, the mixture was purged and cooled to 0 °C. 679.70 mg of iodine (2.68 mmol) was added under light-protected conditions, and the reaction was continued at 0 °C for 30 min. 197.40 mg of compound B19 (1.07 mmol) was dissolved in 5 mL of dry dichloromethane and added to the above reaction flask. The mixture was then allowed to react at room temperature for 4 h. The reaction was monitored by TLC until complete. Most of the dichloromethane was removed by rotary evaporation. 30 mL of diethyl ether was added, and the mixture was washed three times with saturated sodium thiosulfate solution, once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 50:1) to obtain product B20 332.70 mg, a colorless oil, with a yield of 76.86%.

[0137] (5) Preparation of compound B21

[0138] 332.70 mg B20 (0.82 mmol) and 573.90 mg B10 (4.94 mmol) were dissolved in 25 mL of dry tetrahydrofuran under nitrogen protection, ventilated, and cooled to -78 °C. 2.47 mL of lithium diisopropylaminol LDA (2.0 M 4.94 mmol) was added dropwise, and the solution changed from colorless to yellow. The reaction was allowed to proceed overnight from -78 °C to room temperature. The reaction was monitored by TLC until complete. 40 mL of saturated ammonium chloride solution was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 20:1) to give 138.90 mg of product B21, a colorless oil, with a yield of 44.33%.

[0139] (6) Preparation of compound M13

[0140] 138.90 mg B11 (0.37 mmol) was dissolved in 10 mL of ethanol, and 180 mg potassium hydroxide was dissolved in 1 mL of water. These solutions were added to the reaction flask, and the mixture was refluxed overnight. The reaction was monitored by TLC until complete. The ethanol was removed by rotary evaporation, and the mixture was diluted with 10 mL of water. The pH was adjusted to approximately 2 with 1 M hydrochloric acid, resulting in the precipitation of a solid. The solid was extracted three times with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (DCM:MeOH = 50:1) to obtain 105.90 mg of product M10 as a white solid, with a yield of 89.43%. 1 H NMR (400MHz, CDCl3) δ6.28–6.19(m,2H),5.53–5.36(m,2H),2.22–2.08(m,4 H),1.61–1.48(m,4H),1.43–1.34(m,4H),1.38–1.21(m,2H),1.18(s,12H).

[0141] Compound M13 has a ZZ configuration.

[0142] By replacing different substrates and using a synthetic route similar to that of M13, the following compounds were obtained.

[0143] Compound M14: 1 H NMR (400MHz, CDCl3) δ6.27 (dd, J=15.0, 11.0Hz, 1H), 5.95 (dd, J=10.9, 10.9Hz, 1H),5.82(tq,J=4.2,2.0Hz,2H),5.72(tt,J=5.6,2.5Hz,2H),5.58(dt,J=14.7 ,7.1Hz,1H),5.31(dt,J=10.8,7.9Hz,1H),2.39(q,J=7.0Hz,6H),2.12(dq,J=1 0.4, 6.2Hz, 4H), 1.76 (dt, J=14.4, 6.5Hz, 4H), 1.65–1.50 (m, 2H), 1.23 (s, 12H).

[0144] Compound M15: 1 H NMR (400MHz, CDCl3) δ6.30–6.18(m,2H),5.52–5.41(m,2H),2.16(q,J=7.3Hz,4H),1.56(dt,J=9.5,4.2Hz,4H),1.43–1.29(m,4H),1.19(s,12H).

[0145] Compound M16: 1H NMR (400MHz, CDCl3) δ6.32–6.20(m,2H),5.40–5.31(m,2H),2.18(q,J=6.4Hz,4H),1.48(dd,J=10.6,6.1Hz,4H),1.36(p,J=6.5Hz,4H),1.17(s,16H).

[0146] Compound M17: 1 H NMR (400MHz, CDCl3) δ6.26–6.20(m,2H),5.48–5.36(m,2H),2.17(p,J=6.7Hz,4H),1 .58–1.49(m,4H),1.39(q,J=7.4Hz,4H),1.29(tt,J=9.7,5.5Hz,6H),1.18(s,12H).

[0147] Compound M18: 1 H NMR (400MHz, CDCl3) δ6.28–6.17(m,2H),5.48–5.37(m,2H),2.15(q,J=7.1Hz,4H),1.52 (t,J=7.4Hz,4H),1.40(t,J=7.1Hz,4H),1.27(d,J=11.7Hz,8H),1.19(d,J=4.1Hz,12H).

[0148] Compound M19: 1 H NMR (400MHz, CDCl3) δ6.30–6.17(m,2H),5.51–5.39(m,2H),2.15(q,J=7.0Hz,4H),1 .53(d,J=10.3Hz,4H),1.36(t,J=7.3Hz,4H),1.27(d,J=9.9Hz,10H),1.18(s,12H).

[0149] Compound M20: 1 H NMR (400MHz, CDCl3) δ6.30–6.21(m,2H),5.42(dt,J=10.2,7.5Hz,2H),2.16(q,J= 7.1Hz,4H),1.51(t,J=7.4Hz,4H),1.40–1.31(m,4H),1.25(s,12H),1.18(s,12H).

[0150] Compound M21: 1H NMR (400MHz, CDCl3) δ6.29–6.19(m,2H),5.48–5.37(m,2H),2.22–2.10(m,4H), 1.52(t,J=7.2Hz,4H),1.36(d,J=9.6Hz,4H),1.31–1.21(m,14H),1.18(s,12H).

[0151] Compound M22: 1 H NMR (400MHz, CDCl3) δ6.27–6.20(m,2H),5.48–5.38(m,2H),2.16(q,J=7.2Hz,4 H),1.52(t,J=7.2Hz,4H),1.42–1.34(m,4H),1.31–1.23(m,16H),1.18(s,12H).

[0152] Compound M23: 1 H NMR (400MHz, CDCl3) δ6.29–6.19(m,2H),5.48–5.39(m,2H),2.16(q,J=7.2Hz, 4H),1.52(t,J=7.3Hz,4H),1.37(t,J=7.3Hz,4H),1.26(s,18H),1.18(s,12H).

[0153] Compound M24: 1 H NMR (400MHz, CDCl3) δ6.31–6.19(m,2H),5.43(q,J=8.3Hz,2H),2.16(q,J=7.1Hz ,4H),1.52(t,J=7.2Hz,4H),1.37(t,J=7.3Hz,4H),1.26(s,20H),1.18(s,12H).

[0154] Compound M25: 1 H NMR (400MHz, CDCl3) δ6.32–6.20(m,2H),5.82(dq,J=5.3,1.8Hz,2H),5.72(dt,J=5.7,1.9Hz,2H),5.50–5.35(m,2H),2.39(dddt ,J=10.1,8.1,5.5,2.2Hz,6H),2.16(q,J=7.2Hz,4H),1.84–1.69(m,2H),1.63–1.53(m,2H),1.36(q,J=6.8Hz,4H),1.26(s,22H).

[0155] Table 1. Compound Structures

[0156] Test Example 1: ACLY Enzyme Activity Test

[0157] 1.1 Instruments, Materials and Principles

[0158] Instrument: Envision (PerkinElmer, USA)

[0159] Materials: Acl—human ACLY / acly / ATP citrate lyase protein (His tag) purchased from Sino Biological; kinase assay kit ADP-GLO (Promega #V9102).

[0160] Experimental Principle: In this experiment, the ATP-dependent citrate lyase ACL catalyzes the conversion of citrate to acetyl-CoA, which in turn produces malonyl-CoA, a precursor molecule for fatty acid synthesis. This reaction is accompanied by ATP consumption; therefore, the ADP-Glo ​​Kinase Assay (#V9102) can be used to detect changes in ATP, indirectly reflecting the inhibitory effect of the compound on the ACL enzyme activity. This kinase assay kit can quantitatively determine the amount of ADP produced by the ACL enzyme activity. (First, the compound, enzyme, and substrate are co-incubated at 37°C for half an hour. Then, the ADP-Glo ​​AK reagent is added and incubated for another half hour to terminate the reaction and consume the remaining ATP. Next, the kinase assay reagent is added (which converts ADP to ATP while simultaneously using a coupled luciferase / luciferin reaction to detect the newly synthesized ATP) and incubated for another half hour. The value is read using Envision.)

[0161] 1.2 Methods

[0162] The ADP-Glo ​​luminescence assay was used for determination. It quantitatively detects the amount of ADP to reflect the activity of the ACLY enzyme; the amount of ADP detected by the ACLY-catalyzed enzymatic reaction is proportional to the amount of ADP detected by the luminescence signal. First, the compound was diluted in 10% DMSO. Then, 1 μL of the diluted compound was added to 5 μL of the reaction mixture, resulting in a final DMSO concentration of 2%. The ACLY-catalyzed enzymatic reaction was carried out at 37°C for 30 min. The 5 μL reaction mixture contained the following components: 40 mM Tris, pH 8.0, 10 mM MgCl2, 5 mM DTT, ATP, CoA, and Sodium Citrate and ACLY. After the enzymatic reaction, 2.5 μL of ADP-Glo ​​reagent was added to each reaction mixture and incubated at room temperature for 1 hour. Then, 5 μL of the kinase detection reagent was added and incubated at room temperature for 30 minutes. The luminescence signal was detected using Envision (PerkinElmer, USA).

[0163] Data processing: For test activity dose dependence of compounds, IC 50 The value is obtained by nonlinearly fitting the sample activity to the sample concentration. The calculation software used is Graphpad Prism 7, and the fitting model is a sigmoidal dose-response (variable slope). For most inhibitor screening models, the bottom and top of the fitted curve are set to 0 and 100, respectively. Generally, each sample is tested in duplicate (n≥2), and the results are expressed as standard deviation (SD) or standard error (SE).

[0164] 1.3 Results

[0165] The experimental results are shown in Table 2 below. NDI-091143 is a reported ACLY inhibitor and was used as a positive control.

[0166] Table 2 shows the inhibitory activity of the compounds on ACLY in the examples.

[0167] Note: NA indicates no inhibitory activity; * indicates that the inhibitory activity of this compound on lipid synthesis was not measured.

[0168] 326E is a reported AMPK activator and was used as a positive control. Its structural formula is as follows:

[0169] NDI-091143 is a human ATP-citrate lyase (ACLY) inhibitor. In this invention, NDI-091143 was purchased from Bider Pharmaceuticals. Its structural formula is as follows:

[0170] Test Example 2: Hepatic Primary Cell Lipid Synthesis Inhibition Activity Test

[0171] 2.1 Materials, Instruments and Principles

[0172] Materials and instruments: primary liver cells, C14-labeled sodium acetate, PBS, HG-DMEM medium, potassium hydroxide, methanol, chloroform, scintillation solution, 12-well cell plates, isotope detector, cell culture incubator.

[0173] Experimental Principle: Acetyl-CoA, produced by the cleavage of citrate by ACLY, is a fundamental module in lipid synthesis. After primary mouse hepatocytes have been starved and depleted of their intracellular energy reserves, insulin stimulation promotes the absorption of extracellular glucose for cellular utilization and enhances de novo lipid synthesis. C14-labeled sodium acetate is converted intracellularly into isotope-labeled acetyl-CoA. The lipids involved in synthesis can also be isotopically labeled, allowing for the quantitative determination of cellular lipid synthesis activity.

[0174] 2.2 Methods

[0175] Gelatin was deposited in 12-well plates, and primary mouse hepatocytes (300,000 / mL) were seeded into the plates. After 6 hours, the plates were replaced with serum-free HG-DMEM. The next day, the test compound was added to treat the cells, and 10 nM insulin was added to stimulate lipid synthesis. 975 μL of substrate C14-labeled sodium acetate solution was added, and the plates were incubated for 4 hours. Each well was washed three times with 1 mL of PBS, and the plates were dried. 600 μL of KOH solution was added to each well to lyse the cells for 1 hour. 480 μL of the lysate was transferred to a 2 mL EP tube, and 1 mL of extraction buffer (chloroform:methanol (v:v) = 2:1) was added. The plates were then rotated for 10 hours. The remaining lysate was used for protein concentration determination. The suspension was centrifuged at 2500g for 10 min, and the lower layer (chloroform layer) was transferred to a new 2 mL EP tube. Place the tube in a fume hood to evaporate the chloroform, add 1000 μL of scintillation fluid to each tube, and read the values.

[0176] 2.3 Results

[0177] The experimental results are shown in Table 3 below.

[0178] Table 3 shows the inhibitory activity of the compounds in the examples on lipid synthesis.

[0179] Note: ACLY participates in lipid synthesis in vivo by regulating acetyl-CoA. The inhibitory effect of the compound on lipid synthesis is used to reflect the indirect inhibitory effect of the compound on ACLY; * indicates that the inhibitory activity of the compound on lipid synthesis was not measured.

[0180] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A compound represented by general formula I, or a stereoisomer, enantiomer, diastereomer, racemate, or pharmaceutically acceptable salt thereof, In the formula, Each double bond can be independently configured in either cis (Z) or trans (E) configuration; R1 and R2 are each independently H, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C7 cycloalkyl, C3-C7 cycloalkenyl, C6 or C10 aryl; or R1 and R2 together with the connected carbon form a saturated 3-7 membered ring, or a 3-7 membered ring containing an unsaturated double bond; R3 and R4 are each independently H, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C7 cycloalkyl, C3-C7 cycloalkenyl, C6 or C10 aryl; or R3 and R4 together with the connected carbon form a saturated 3-7 membered ring, or a 3-7 membered ring containing an unsaturated double bond; m, n, and q are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9; The aforementioned C1-C4 alkyl, C2-C4 alkenyl, C2-C4 ynyl, C3-C7 cycloalkyl, C3-C7 cycloalkenyl, or C6-C10 aryl groups are either unsubstituted or substituted, wherein... The substitution refers to substitution by 1, 2, 3, 4 or 5 substituents selected from the group consisting of: halogen, C1-C4 alkyl, C1-C4 alkoxy, C3-C7 cycloalkyl, C3-C7 cycloalkenyl, and C6-C10 aryl.

2. The compound of claim 1, or its stereoisomers, enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts thereof, characterized in that, R1 and R2 are each independently H, methyl, ethyl, propyl, vinyl, propenyl, ethynyl, propynyl, C3-C6 cycloalkyl, C3-C6 cycloalkenyl, or C6-C10 aryl; or, R1 and R2 together with the connected carbon form a saturated 3-6 membered ring, or a 3-6 membered ring containing one unsaturated double bond.

3. The compound of claim 1, or its stereoisomers, enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts thereof, characterized in that, R3 and R4 are each independently H, methyl, ethyl, propyl, vinyl, propenyl, ethynyl, propynyl, C3-C6 cycloalkyl, C3-C6 cycloalkenyl, or C6-C10 aryl; or, R3 and R4 together with the connected carbon form a saturated 3-6 membered ring, or a 3-6 membered ring containing one unsaturated double bond.

4. The compound of claim 1, or its stereoisomers, enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts thereof, characterized in that, R1 and R2 together with the attached carbon form a saturated four-membered ring, a saturated five-membered ring, a saturated six-membered ring, a four-membered ring containing one unsaturated double bond, a five-membered ring containing one unsaturated double bond, or a six-membered ring containing one unsaturated double bond.

5. The compound of claim 1, or its stereoisomers, enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts thereof, characterized in that, R3 and R3 together with the attached carbon form saturated four-membered rings, saturated five-membered rings, saturated six-membered rings, four-membered rings containing one unsaturated double bond, five-membered rings containing one unsaturated double bond, or six-membered rings containing one unsaturated double bond.

6. The compound of claim 1, or its stereoisomers, enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts thereof, characterized in that, The compound has the structure shown in Formula II: Where m, n and q are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8 or 9.

7. The compound as claimed in claim 1 or 6, or its stereoisomers, enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts thereof, characterized in that, q = 0, 1, 2 or 3.

8. The compound as claimed in claim 1 or 6, or its stereoisomers, enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts thereof, characterized in that, m and n are each independently 4, 5, 6, 7, 8 or 9.

9. The compound as claimed in claim 1 or 6, or its stereoisomers, enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts thereof, characterized in that, The two double bonds are either in cis (Z) or trans (E) configurations, each independent of the other.

10. The compound of claim 1, or its stereoisomers, enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts thereof, characterized in that, The compounds are selected from the group consisting of:

11. A pharmaceutical composition, characterized in that, Include: The compound as claimed in any one of claims 1-10, or a stereoisomer, enantiomer, diastereomer, racemate, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

12. The use of the compound according to any one of claims 1-10, or its stereoisomers, enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts thereof, or the pharmaceutical composition according to claim 11, characterized in that, Used for the preparation of ACLY inhibitors; or for the preparation of drugs for the prevention and / or treatment of metabolic diseases or cancer.

13. The use as described in claim 12, characterized in that, The metabolic diseases mentioned are selected from the following group: hypercholesterolemia, atherosclerosis, non-alcoholic fatty liver disease, and diabetes.

14. The use as described in claim 12, characterized in that, The cancers mentioned are selected from the following group: lung cancer, pancreatic cancer, breast cancer, ovarian cancer, liver cancer, colorectal cancer, brain cancer, and acute myeloid leukemia.

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