A tricyclic fxr receptor antagonist and a method of making the same

Abstract

The present application relates to a kind of tricyclic FXR antagonists and its preparation method, the structure of the tricyclic FXR antagonist is as shown in general formula I, the definition of each substituent group is as described in the specification and claims, compound as shown in general formula I can be used as FXR antagonist, can be used to prepare the drug for treating cholestasis, hyperlipidemia, diabetes and the like.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry, and specifically relates to a tricyclic FXR receptor antagonist, its preparation method, and the use of this type of FXR antagonist in FXR antagonistic activity. Background Technology

[0002] The FXR receptor, a member of the nuclear hormone receptor superfamily, plays a central regulatory role in bile acid synthesis and enterohepatic circulation, thereby maintaining bile acid homeostasis. In the liver, bile acids activate FXR, inducing the expression of the small heterodimer chaperone SHP, inhibiting the transcription of genes for two enzymes in the bile acid synthesis pathway, CYP7A1 and CYP8B1, thus reducing bile acid synthesis. FXR can also induce the expression of the bile salt efflux pump protein BESP, facilitating the excretion of bile acids into the bile ducts. In intestinal epithelial cells, FXR receptor activation by bile acids induces the expression of FGF19 (FGF15 in mice). FGF19 / 15 reaches the liver via the portal vein, binds to the FGF4 / β-Klotho heterodimer complex in hepatocytes, initiates the ERK1 / 2 cascade signaling pathway, and inhibits the expression of the CYP7A1 rate-limiting enzyme. FXR receptor can also induce the expression of ileal bile acid-binding protein (IBABP), promoting bile acid efflux. The FXR signaling pathway is also closely related to lipid and carbohydrate metabolism, with the liver and intestines being central organs for FXR-dependent carbohydrate and lipid metabolism. Furthermore, FXR receptors are also closely associated with cholestasis, liver cancer, and gut health.

[0003] Although there are currently no FXR receptor antagonists used for clinical treatment, they have been developed as important research tools. Some compounds have also shown effectiveness in treating cholestasis and hypercholesterolemia in animal models. For example, to determine whether bisabolol, as an FXR antagonist, has a cholesterol-lowering effect, wild-type mice and FXR gene-deficient mice were fed a high-cholesterol diet. Both wild-type and FXR-deficient mice showed increased liver cholesterol levels. Oral administration of bisabolol reduced liver cholesterol levels in wild-type mice, but not in FXR gene-deficient mice. Oral administration of Gly-MCA to mice can prevent and treat diet-induced obesity and hereditary obesity, improve insulin resistance and hepatic steatosis, without systemic, hepatic, or intestinal toxicity. Jiang discovered that GUDCA is an endogenous FXR receptor antagonist that can be degraded by bile acid hydrolases. Metformin reduces the activity of bile acid hydrolases by decreasing Bacteroides fragilis in the gut, thereby increasing GUDCA levels in the gut, which inhibits FXR transcription, ultimately lowering blood sugar. When mice were treated with tempol or antibiotics, the number of lactobacilli in the gut was significantly reduced, leading to decreased levels of bile acid hydrolase BSH and increased T-β-MCA content in the ileum, thereby inhibiting FXR signaling. Inhibition of intestinal FXR signaling resulted in improved mitochondrial function and inhibition of ceramide synthesis, leading to decreased serum ceramide levels. Reduced circulating ceramide downregulated the expression of hepatic SREBP1C and CIDEA, thereby reducing hepatic steatosis. The natural FXR antagonist isoDCA can attenuate immunostimulation by antagonizing FXR receptors in dendritic cells (DCs), increasing Foxp3 induction and increasing intestinal CD4 levels. + The number of regulatory T cells, in turn, affects gut health. Therefore, FXR antagonists offer new avenues for the treatment of some metabolic diseases.

[0004] Reported FXR antagonists are mainly divided into two categories: natural product antagonists and synthetic small molecule antagonists. Guggulaterone (GS) was the first reported FXR receptor antagonist. In 2019, Daniela Passeri et al. reported the biological activity of GS on the FXR receptor. The authors observed that GS could activate LXRβ, PXR, and PR receptors at micromolar concentrations, while it failed to induce activation of other nuclear receptors, including FXR. In antagonism assays, GS reduced the FXR agonistic activity of CDCA, with an IC50 value of [missing value]. 50 The concentration was 25 μM. The IC50 for other nuclear receptors was... 50Values ​​ranged from 0.32 to 62 μM (Med. Chem. Comm. 2019, 10, 8). A recently reported nonsteroidal FXR receptor antagonist, FLG249, retains strong antagonistic activity while also exhibiting metabolic stability. In tissue distribution experiments, it was primarily concentrated in the ileum of mice. Oral administration of FLG249 downregulated the mRNA levels of FXR target genes Fgf15, Asbt, and Shp in the mouse ileum (ACSMed. Chem. Lett. 2021, 12, 420).

[0005]

[0006] Currently available FXR antagonists have limited natural sources and are difficult to synthesize. There are few structural modifications to these compounds, and further research is needed. Summary of the Invention

[0007] The object of this invention is to provide a compound that can be used as a farnesoid derivative X receptor (FXR) antagonist.

[0008] In a first aspect, a compound of general formula (I), or a pharmaceutically acceptable salt thereof,

[0009]

[0010] R1 is hydrogen, hydroxyl, halogen, or C1-C6 alkyl;

[0011] R2 is selected from: hydrogen, hydroxyl group, halogen, C1-C6 alkoxy group, 3- to 10-membered cycloalkoxy group, =O, =N-OH, R d -C(=O)-O-; where R d The following groups, substituted or unsubstituted, may be used: C1-C8 alkyl, C3-C10 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl, R a NH- or R a O-; where each R a Independently selected from: C1-C8 alkyl, C3-C10 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl;

[0012] R3 is an unsubstituted or substituted C1-C6 alkyl group, or hydrogen;

[0013] R4 is absent, hydrogen, hydroxyl, halogen, or C1-C6 alkyl;

[0014] R5 is selected from: hydrogen, hydroxyl group, halogen, C1-C6 alkoxy group, 3- to 10-membered cycloalkoxy group, =O, =N-OH, R e -C(=O)-O- or O=C(R) e )-O-; where each R eThe following groups are independently hydrogen or substituted or unsubstituted: C1-C8 alkyl, C3-C10 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl, R b NH- or R b O-; where each R b Independently selected from: C1-C8 alkyl, C3-C10 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl;

[0015] R6 is selected from hydrogen, hydroxyl, hydroxymethyl, formyl, ... -(C1-C6 alkylene)C(=O)-R f -R g Where X is NH, O, or S; R c It is hydrogen, unsubstituted or substituted C1-C6 alkyl; R f It is O, S or NH; Rg is hydrogen, substituted or unsubstituted C1-C6 alkyl;

[0016] Each * independently represents the R configuration, S configuration, or racemic configuration;

[0017] The substitution refers to the substitution of hydrogen on the group by one or more substituents selected from the group consisting of: hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, carboxyl (-COOH), and sulfonic acid (-SO2OH).

[0018] In another preferred embodiment, R1 is hydrogen.

[0019] In another preferred embodiment, R2 is a hydroxyl group or R d -C(=O)-O-; where R d The following groups, substituted or unsubstituted, may be used: C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, 5-7 heteroaryl, R a NH- or R a O-; where each R a Independently selected from C1-C6 alkyl groups;

[0020] The substitution refers to the substitution of hydrogen on the group by one or more substituents selected from the group consisting of halogens, C1-C6 alkyl groups, and C1-C6 alkoxy groups.

[0021] In another preferred embodiment, R2 is connected to the ring in an α configuration.

[0022] In another preferred embodiment, R3 is hydrogen or a C1-C4 alkyl group.

[0023] In another preferred embodiment, R4 is absent, and R5 is either =O or =N-OH;

[0024] Or R4 is hydrogen, R5 is a hydroxyl group, halogen, C1-C4 alkoxy group, 3- to 8-membered cycloalkoxy group, Re -C(=O)-O- or O=C(R) e )-O-; where each R e The following groups are independently hydrogen or substituted or unsubstituted: C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl, R b NH- or R b O-; where each R b The group is independently selected from: C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, 5-7 heteroaryl; the substitution refers to the substitution of the hydrogen on the group by one or more substituents selected from the group consisting of: hydroxyl, halogen, C1-C4 alkyl, C1-C4 alkoxy, carboxyl.

[0025] In another preferred embodiment, R6 is selected from: hydroxyl, hydroxymethyl, formyl, ... -(C1-C6 alkylene)C(=O)-R f -R g Where X is NH or O; R c It is hydrogen, unsubstituted or substituted C1-C4 alkyl; R f It is O or NH; Rg is hydrogen, substituted or unsubstituted C1-C4 alkyl;

[0026] The substitution refers to the substitution of hydrogen on the group by one or more substituents selected from the group consisting of C1-C4 alkyl, carboxyl (-COOH), and sulfonic acid (-SO2OH).

[0027] In another preferred embodiment, the compound is:

[0028]

[0029]

[0030] A second aspect of the present invention provides a method for preparing the compound described in the first aspect or a pharmaceutically acceptable salt thereof, the method using cholesterol as a raw material, by transferring a double bond to a five-membered ring through a chemical reaction, then obtaining a compound containing a carboxylic acid side chain through oxidative ring opening, and then introducing an amino acid group onto the carboxyl group to obtain the compound described in the first aspect, wherein the definitions of each substituent are as described in the first aspect.

[0031] In another preferred embodiment, the compound structure is as shown in formula H3 or H4, and it is prepared via the following route:

[0032]

[0033] The reaction of compound H1 with acyl chloride and acid anhydride yields compound H2;

[0034] Compound H2 is debenzyl protecting group to give compound H3;

[0035] The reaction of compound H3 with amino acids or chloroformates yields compound H4.

[0036] Where R7 is a substituted or unsubstituted group of the following: C1-C8 alkyl, C3-C10 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl, R a NH- or R a O-; where each R a Independently selected from: C1-C8 alkyl, C3-C10 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl;

[0037] A is either O or NH;

[0038] R8 is selected from hydrogen, unsubstituted or substituted C1-C6 alkyl; the substitution means that the hydrogen on the group is replaced by one or more substituents selected from the group consisting of: hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, carboxyl (-COOH), sulfonic acid (-SO2OH).

[0039] A third aspect of the present invention provides a pharmaceutical composition comprising the compound described in the first aspect or a pharmaceutically acceptable salt thereof;

[0040] And pharmaceutically acceptable carriers.

[0041] A fourth aspect of the invention provides the use of the compound described in the first aspect or a pharmaceutically acceptable salt thereof or a pharmaceutical composition described in the third aspect, characterized in that: (i) it is used to prepare a farnesoid derivative X receptor (FXR) antagonist; (ii) it is used to prepare a medicament for treating diseases associated with farnesoid derivative X receptors; or (iii) it is used to prepare a medicament for treating metabolic diseases.

[0042] In another preferred embodiment, the disease associated with the farnesoid derivative X receptor is selected from: hyperlipidemia, bile acid stasis, diabetes, obesity, non-alcoholic fatty liver disease, biliary cirrhosis, and hypercholesterolemia.

[0043] In another preferred embodiment, the metabolic disease is selected from: hyperlipidemia, bile acid stasis, diabetes, obesity, non-alcoholic fatty liver disease, biliary cirrhosis, and hypercholesterolemia.

[0044] The compounds of this invention can effectively antagonize FXR receptors at micromolar concentrations, and have richer natural sources and simpler synthetic methods compared to some existing naturally derived FXR antagonists.

[0045] 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. Each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Due to space limitations, they will not be described in detail here. Detailed implementation method:

[0046] Through extensive and in-depth research, the inventors of this application have, for the first time, developed a class of compounds designed based on a cholesterol core. Their main characteristic is the retention of the 3-position hydroxyl configuration and the formation of a carboxylic acid side chain through D-ring opening, with the introduction of glycine or taurine groups at this position, resulting in significant FXR antagonistic activity. The inventors have also, for the first time, studied the influence of this structure on activity, obtaining a series of compounds with excellent performance. Compared with some existing naturally derived FXR antagonists, these compounds exhibit comparable activity, and not only are cholesterol sources more abundant and inexpensive, but their synthetic routes are also simpler and yields are ideal, making them promising novel drugs for treating metabolic diseases targeting this target. Based on this, the present invention was completed.

[0047] the term

[0048] In this document, expressions in the form "C1-C8" are intended to include corresponding groups having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms. For example, "C1-C8 alkyl" refers to an alkyl group having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, and "C2-C10 alkenyl" refers to an alkenyl group having 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms.

[0049] In this document, the alkyl group is preferably an aliphatic alkyl group, which may be a straight-chain alkyl group or a branched alkyl group, and includes, without limitation, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, etc.

[0050] In this document, the alkenyl group is preferably vinyl, propenyl, butenyl, styrene, styrene, or similar.

[0051] In this document, alkoxy refers to -O-(alkyl), where alkyl is defined as described above. "C1-6 alkoxy" refers to alkyloxy groups containing 1-6 carbons, and non-limiting examples include methoxy, ethoxy, propoxy, butoxy, etc.

[0052] In this document, the cycloalkyl group can be a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent comprising 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, and more preferably 3 to 10 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentenyl, cyclohexyl, cyclooctyl, etc.; polycyclic cycloalkyl groups include spirocyclic, fused-ring, and bridged-ring cycloalkyl groups.

[0053] In this article, cycloalkoxy refers to -O- (cycloalkyl), where cycloalkyl is defined as described above.

[0054] The aryl group refers to a 6- to 10-membered all-carbon monocyclic or fused polycyclic (i.e., a ring sharing adjacent carbon atom pairs) group, and the group has a conjugated π-electron system, such as phenyl and naphthyl. The aryl ring may be fused with a heterocyclic, heteroaryl, or cycloalkyl ring, and non-limiting examples include benzimidazole, benzothiazole, benzoxazole, benzoisoxazole, benzopyrazole, quinoline, benzoindole, and benzodihydrofuran.

[0055] The heteroaryl group refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms include oxygen, sulfur, and nitrogen. The heteroaryl group is preferably 5- or 6-membered, such as furanyl, thiophene, pyridinyl, pyrroleyl, N-alkylpyrroleyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl group can be fused to an aryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring.

[0056] Unless otherwise indicated in this invention, Indicates the connection site.

[0057] Unless otherwise specified, the structural formulas described in this invention are intended to include all tautomers, optical isomers, and stereoisomers (e.g., enantiomers, diastereomers, geometric isomers, or conformational isomers): for example, R, S configurations containing an asymmetric center, (Z), (E) isomers of double bonds, and (Z), (E) conformational isomers. Therefore, any single stereochemical isomer, tautomer, or enantiomer, diastereomer, geometric isomer, conformational isomer, or mixture of tautomers of the compounds of this invention is within the scope of this invention.

[0058] The term "tautomer" refers to structural isomers with different energies that can cross a low energy barrier and thus interconvert. For example, proton tautomers (i.e., proton shifts) include interconversions via proton migration, such as 1H-indazole and 2H-indazole, 1H-benzo[d]imidazole and 3H-benzo[d]imidazole. Valence tautomers include interconversions via some bonding electron recombination.

[0059] In this document, there are no particular limitations on the pharmaceutically acceptable salts, but they preferably include: inorganic acid salts, organic acid salts, alkyl sulfonates, and aryl sulfonates; the inorganic acid salts include hydrochlorides, hydrobroms, nitrates, sulfates, phosphates, etc.; the organic acid salts include formates, acetates, propionates, benzoates, maleates, fumarates, succinates, tartrates, citrates, etc.; the alkyl sulfonates include methyl sulfonates, ethyl sulfonates, etc.; and the aryl sulfonates include benzene sulfonates, p-toluene sulfonates, etc.

[0060] Preparation method

[0061] The compounds of the present invention can be prepared by the following route.

[0062] Route 1:

[0063]

[0064] Route 2

[0065] The definition of R in each compound is as follows:

[0066] Route 3

[0067]

[0068]

[0069] Route 4

[0070]

[0071]

[0072]

[0073] 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 weight percentages and parts by weight.

[0074] 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.

[0075] In the following preparation examples, NMR was measured using a Varian Mercury-Vx 300M instrument, with NMR calibration: δH 7.26 ppm (CDCl3), 2.50 ppm (DMSO-d6); mass spectrometry was performed using an Agilent 1200 Quadrupole LC / MS system or a SHIMADZU GCMS-QP5050A; reagents were mainly provided by Shanghai Chemical Reagent Company; TLC silica gel plates were manufactured by Shandong Yantai Huiyou Silica Gel Development Co., Ltd., model HSGF 254; and 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.

[0076] The Chinese translations of the abbreviations in this article are as follows:

[0077] DMAP: 4-Dimethylaminopyridine; DCM: Dichloromethane; DMF: N,N-Dimethylformamide; TFA: Trifluoroacetic acid.

[0078] Example 1

[0079]

[0080] (1) Cholesterol S-1 (10 g, 25.91 mmol) and a catalytic amount of DMAP (31.65 mg, 0.259 mmol) were dissolved in 120 mL of dichloromethane. Pyridine (20 mL) was added, and acetic anhydride (3.65 mL, 38.86 mmol) was added dropwise under an ice-water bath. The mixture was stirred at room temperature for 4 h. TLC monitoring showed that the reaction was complete. The dichloromethane was removed by evaporation, and the pyridine was neutralized with 1 N dilute hydrochloric acid until the solution was neutral. The mixture was extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with deionized water and saturated brine, dried over sodium sulfate and concentrated. The intermediate S-2 was separated by column chromatography to obtain 10.56 g (24.67 mmol), with a molar yield of 95.2%. 1 ¹H NMR (400MHz, Chloroform-d) δ 5.37 (d, J = 5.0 Hz, 1H), 4.64–4.56 (m, 1H), 2.31 (d, J = 8.0 Hz, 2H), 2.03 (s, 3H), 2.00–1.06 (m, other alicyclic protons), 1.02 (s, 3H), 0.91 (d, J = 6.4 Hz, 3H), 0.87–0.85 (m, 6H), 0.68 (s, 3H).

[0081] (2) The product S-2 (10 g, 23.36 mmol) from the previous step was dissolved in cyclohexane (100 mL), ruthenium trichloride (48.45 mg, 0.234 mmol) was added, and tert-butyl hydroperoxide (5.84 mL, 58.4 mmol) was added dropwise. The reaction was carried out at room temperature for 7 h. When the reaction was complete as monitored by TLC, the reaction was quenched with a saturated aqueous solution of sodium bisulfite (50 mL), and then extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with deionized water and saturated brine, respectively, dried over sodium sulfate, concentrated, and separated by column chromatography to obtain intermediate S-3 5.37 g (12.15 mmol), with a molar yield of 52.0%. 1 ¹H NMR (400MHz, Chloroform-d) δ 5.70 (s, 1H), 4.75–4.67 (m, 1H), 2.57–2.36 (m, 3H), 2.23 (t, J = 10.8 Hz, 1H), 2.05 (s, 3H), 2.03–1.22 (m, other alicyclic protons), 1.20 (s, 3H), 1.17–0.99 (m, other alicyclic protons), 0.92 (d, J = 6.4 Hz, 3H), 0.86 (d, J = 6.4 Hz, 6H), 0.68 (s, 3H).

[0082] (3) The product S-3 (5 g, 11.31 mmol) from the previous step was dissolved in dichloromethane (50 mL). After changing the nitrogen atmosphere, 10% Pd / C (50 mg) was quickly added. After changing the nitrogen atmosphere again, hydrogen atmosphere was added, and the mixture was stirred at room temperature. When the reaction was complete as monitored by TLC, the nitrogen atmosphere was changed again, the Pd / C was filtered off, the reaction solution was evaporated to dryness, and then separated by column chromatography to obtain intermediate S-4 4.03 g (9.08 mmol), with a molar yield of 80.25%. 1 ¹H NMR (400MHz, Chloroform-d) δ 4.70–4.64 (m, 1H), 2.33 (t, J = 12.0 Hz, 2H), 2.19 (d, J = 9.6 Hz, 1H), 2.02 (s, 3H), 1.93–1.21 (m, other alicyclic protons), 1.09 (s, 3H), 0.90 (d, J = 6.5 Hz, 3H), 0.87–0.85 (m, 6H), 0.65 (s, 3H).

[0083] (4) Dissolve the product S-4 (4 g, 9.01 mmol) from the previous step in a 1:1 volume mixture of tetrahydrofuran and methanol, add cerium trichloride (2.44 g, 9.91 mmol), dissolve, and then add sodium borohydride (374.89 mg, 9.91 mmol). React at room temperature. When the reaction is complete as monitored by TLC, quench the reaction with 2N HCl, extract with ethyl acetate (3 × 100 mL), wash the combined organic layers with deionized water and saturated brine, dry and concentrate with sodium sulfate, and separate by column chromatography to obtain intermediate S-54.07 g (9.13 mmol), molar yield: 92.1%. 1 ¹H NMR (400MHz, Chloroform-d) δ 4.73–4.62 (m, 1H), 3.39–3.33 (m, 1H), 2.01 (s, 3H), 1.90–0.95 (m, other alicyclic protons), 0.90 (d, J = 6.4Hz, 3H), 0.87–0.84 (m, 9H), 0.67 (s, 3H).

[0084] (5) Dissolve the product S-5 (4 g, 8.97 mmol) from the previous step in toluene (50 mL), add Burgess Reagent (3.22 g, 13.45 mmol), and proceed with the reaction at 110 °C. When the reaction was complete as monitored by TLC, the reaction solution was evaporated to dryness and then subjected to column chromatography to obtain intermediate S-6 2.60 g (6.07 mmol), with a molar yield of 67.7%. 1 ¹H NMR (400MHz, Chloroform-d) δ 5.14 (s, 1H), 4.72–4.66 (m, 1H), 2.03 (s, 3H), 1.86–0.99 (m, other alicyclic protons), 0.92 (d, J = 6.0 Hz, 3H), 0.86 (d, J = 6.4 Hz, 6H), 0.81 (s, 3H), 0.53 (s, 3H).

[0085] (6) Dissolve the product S-6 (2.5 g, 5.84 mmol) from the previous step in a hydrogen chloride-dioxane solution (15 mL, 58.4 mmol), react overnight at room temperature, and then directly evaporate the reaction solution to dryness and column chromatography to obtain intermediate S-7 2.14 g (5.00 mmol), molar yield: 85.6%. 1 ¹H NMR (400MHz, Chloroform-d) δ 5.15 (s, 1H), 4.75–4.65 (m, 1H), 2.40–2.17 (m, 2H), 2.02 (s, 3H), 1.94–1.06 (m, other alicyclic protons), 0.91 (s, 3H), 0.89–0.82 (m, 9H), 0.70 (s, 3H).

[0086] (7) Dissolve the product S-7 (2.0 g, 4.67 mmol) from the previous step in carbon tetrachloride (30 mL), add m-chloroperoxybenzoic acid (967.68 mg, 5.60 mmol), react at room temperature for 4 h, monitor the reaction by TLC until complete, quench the reaction with saturated aqueous sodium bisulfite solution (20 mL), extract with ethyl acetate (3 × 100 mL), wash the combined organic layers with deionized water and saturated brine respectively, dry and concentrate with sodium sulfate, and separate by column chromatography to obtain intermediate S-8 725.72 mg (1.63 mmol), molar yield: 35.0%. 1 ¹H NMR (400MHz, Chloroform-d) δ 4.72–4.65 (m, 1H), 3.30 (s, 1H), 2.08 (m, 2H), 2.01 (s, 3H), 1.86–0.99 (m, other alicyclic protons), 0.97 (s, 3H), 0.85–0.81 (m, 9H), 0.74 (s, 3H).

[0087] (8) Dissolve the product S-8 (700 mg, 1.58 mmol) from the previous step in acetone (20 mL), add Jones' reagent (0.5 mL), and react at room temperature for 0.5 h. Monitor the reaction by TLC until it is complete. After the reaction solution is evaporated to dryness, add 30 mL of water to a round-bottom flask and extract with ethyl acetate (3 × 30 mL). Wash the combined organic layers with deionized water and saturated brine, respectively, dry and concentrate with sodium sulfate, and separate by column chromatography to obtain compound C2 633.08 mg (1.33 mmol), with a molar yield of 84.2%. 1 ¹H NMR (400MHz, DMSO-d6) δ 11.95 (s, 1H), 4.59–4.52 (m, 1H), 2.46 (d, J = 6.9Hz, 1H), 2.17–2.02 (m, 2H), 1.96 (s, 3H), 1.89–1.03 (m, other alicyclic protons), 0.99 (s, 3H), 0.83 (d, J = 6.4Hz, 12H).

[0088] (9) Dissolve the product C2 (600 mg, 1.26 mmol) from the previous step in a mixed solvent of ethanol and water in a volume ratio of 9:1, add potassium hydroxide (705.6 mg, 12.6 mmol), and react at room temperature. Monitor the reaction by TLC until complete, neutralize the reaction solution with 1N dilute hydrochloric acid to acidity, extract with ethyl acetate (3 × 50 mL), wash the combined organic layers with deionized water and saturated brine respectively, dry and concentrate with sodium sulfate, and separate by column chromatography to obtain compound C1 504.73 mg (1.16 mmol), molar yield: 92.3%. 1¹H NMR (400MHz, DMSO-d6) δ 11.93 (s, 1H), 3.37–3.30 (m, 1H), 2.49–2.42 (m, 1H), 2.17–2.01 (m, 2H), 1.95–1.83 (m, 1H), 1.75–1.02 (m, other alicyclic protons), 0.99 (s, 3H), 0.84–0.82 (m, 9H), 0.79 (s, 3H).

[0089] Example 2

[0090]

[0091] (1) Compound C1 (480 mg, 1.11 mmol) was dissolved in DMF (10 mL), and anhydrous potassium carbonate (306.8 mg, 2.22 mmol) was added. Benzyl bromide (1.5 mL, 1.33 mmol) was slowly added dropwise with stirring at room temperature. After the addition was complete, the reaction mixture was transferred to 50 °C and reacted overnight. After the reaction was monitored by TLC until complete, the reaction mixture was cooled to room temperature, diluted with 50 mL of deionized water, and extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with deionized water and saturated brine, respectively, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain intermediate S-9 520 mg (0.992 mmol), molar yield: 86.4%. 1 ¹H NMR (400MHz, Chloroform-d) δ 7.36–7.31 (m, 5H), 5.10 (s, 2H), 3.62–3.57 (m, 1H), 2.60–2.57 (m, 1H), 2.48–2.42 (m, 1H), 2.29–2.27 (m, 2H), 2.01–1.95 (m, 1H), 1.86–1.00 (m, other alicyclic protons), 1.06 (s, 3H), 0.92 (d, J = 6.8Hz, 3H), 0.85 (s, 6H), 0.83 (s, 3H).

[0092] (2) Dissolve the product S-9 (80 mg, 0.153 mmol) and a catalytic amount of DMAP (1.9 mg, 0.0153 mmol) in dichloromethane (10 mL), add pyridine (1 mL), and add propionic anhydride (49.8 mg, 0.382 mmol) under an ice-water bath. After the addition is complete, move the reaction to room temperature and react for 4 h. After the reaction is complete as monitored by TLC, dichloromethane is evaporated, and pyridine is neutralized with 1N dilute hydrochloric acid until the solution is neutral. Extract with ethyl acetate (3 × 20 mL), and wash the combined organic layers with deionized water and saturated brine, respectively. Dry and concentrate with sodium sulfate, and separate by column chromatography to obtain intermediate S-10b 60 mg (0.103 mmol), molar yield: 67.3%.1 ¹H NMR (400MHz, Chloroform-d) δ 7.41–7.29 (m, 5H), 5.10 (s, 2H), 4.72–4.64 (m, 1H), 2.60–2.56 (m, 1H), 2.47–2.40 (m, 1H), 2.31–2.26 (m, 3H), 1.98 (m, 1H), 1.95–1.18 (m, other alicyclic protons), 1.12 (t, J = 7.6Hz, 3H), 1.05 (s, 3H), 0.92 (d, J = 6.8Hz, 3H), 0.89–0.81 (m, 9H).

[0093] (3) Dissolve the product S-10b (60 mg, 0.103 mmol) from the previous step in 10 mL of a mixed solvent of CH3OH / EA = 10:1. After changing the nitrogen atmosphere, quickly add 10% Pd / C (6 mg by mass), then change the nitrogen atmosphere again, followed by hydrogen atmosphere, and stir at room temperature. When the reaction is complete as monitored by TLC, change the nitrogen atmosphere again, filter off the Pd / C, evaporate the reaction solution to dryness, and then perform column chromatography to obtain compound C3 45 mg (0.092 mmol), with a molar yield of 89.2%. 1 ¹H NMR (400MHz, CDCl₃) δ 4.73–4.65 (m, 1H), 2.56–2.52 (m, 1H), 2.50–2.45 (m, 1H), 2.31–2.25 (m, 3H), 1.93–1.20 (m, other alicyclic protons), 1.14–1.10 (m, 6H), 0.95 (d, J = 6.8Hz, 3H), 0.89 (s, 3H), 0.85 (d, J = 6.4Hz, 6H).

[0094] The following compounds were synthesized using different acid anhydrides or acyl chlorides, using the same method as in Example 2.

[0095]

[0096]

[0097]

[0098] Example 3

[0099]

[0100] (1) Compound C2 (87 mg, 0.189 mmol) was dissolved in DMF (10 mL), and anhydrous potassium carbonate (65.3 mg, 0.473 mmol) was added. Benzyl bromoacetate (149 μL, 0.945 mmol) was slowly added dropwise with stirring at room temperature. After the addition was complete, the reaction was allowed to proceed overnight at room temperature. After the reaction was monitored by TLC until complete, the reaction solution was cooled to room temperature, diluted with 50 mL of deionized water, and extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with deionized water and saturated brine, respectively, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain intermediate S-20 106 mg (0.170 mmol), molar yield: 89.9%.

[0101] 1 ¹H NMR (400MHz, Chloroform-d) δ 7.37 (d, J = 1.2 Hz, 5H), 5.24 (s, 2H), 5.18 (s, 2H), 4.63 (d, J = 3.9 Hz, 1H), 2.60 (d, J = 15.9 Hz, 1H), 2.35 (d, J = 6.3 Hz, 2H), 2.02 (s, 3H), 2.00–1.03 (m, other alicyclic protons), 1.14 (s, 3H), 0.95 (d, J = 6.7 Hz, 3H), 0.90–0.83 (m, 9H).

[0102] (2) Dissolve the product S-20 (50 mg, 0.08 mmol) from the previous step in 10 mL of a mixed solvent of CH3OH / EA = 10:1. After changing the nitrogen atmosphere, quickly add 10% Pd / C (5 mg by mass), then change the nitrogen atmosphere again, followed by hydrogen atmosphere, and stir at room temperature. When the reaction is complete as monitored by TLC, change the nitrogen atmosphere again, filter off the Pd / C, evaporate the reaction solution to dryness, and then perform column chromatography to obtain compound C17 40 mg (0.075 mmol), molar yield: 93.6%.

[0103] 1 ¹H NMR (400MHz, Chloroform-d) δ 4.80–4.57 (m, 3H), 2.51 (s, 1H), 2.36–2.33 (m, 2H), 2.02 (s, 3H), 1.95–1.23 (m, other alicyclic protons), 1.14 (s, 3H), 0.95 (d, J = 6.7 Hz, 3H), 0.90–0.83 (m, 9H).

[0104] Example 4

[0105]

[0106] (1) Compound C2 (40 mg, 0.084 mmol), HOBT (11.4 mg, 0.084 mmol), EDCI (16.1 mg, 0.084 mmol), and DMAP (31 mg, 0.0252 mmol) were dissolved in dichloromethane (10 mL). Glycine benzyl ester hydrochloride (25.4 mg, 0.126 mmol) was added, followed by the addition of a catalytic amount of DMF. The reaction was allowed to proceed overnight. After the reaction was completed as monitored by TLC, the reaction solution was evaporated to dryness, and column chromatography was used to separate intermediate S-13 30 mg (0.048 mmol), with a molar yield of 57.3%. 1 ¹H NMR (400MHz, Chloroform-d) δ 7.42–7.29 (m, 5H), 5.18 (s, 2H), 4.72–4.64 (m, 1H), 4.22–4.00 (m, 2H), 2.65–2.58 (m, 1H), 2.38–2.35 (m, 1H), 2.22–2.09 (m, 2H), 2.02 (s, 3H), 1.96–1.19 (m, other alicyclic protons), 1.16 (s, 3H), 0.94 (d, J = 6.8Hz, 3H), 0.90 (s, 3H), 0.85 (d, J = 6.6Hz, 6H).

[0107] (2) Dissolve the product S-13 (30 mg, 0.048 mmol) from the previous step in 10 mL of a mixed solvent of CH3OH / EA = 10:1. After changing the nitrogen atmosphere, quickly add 10% Pd / C (3 mg by mass), then change the nitrogen atmosphere again, followed by hydrogen atmosphere, and stir at room temperature. When the reaction is complete as monitored by TLC, change the nitrogen atmosphere again, filter off the Pd / C, evaporate the reaction solution to dryness, and then perform column chromatography to obtain compound S-14 20 mg (0.0375 mmol), molar yield: 78.2%. 1 ¹H NMR (400MHz, DMSO-d⁶) δ 12.38 (s, ¹H), 8.06 (t, J = 6.0 Hz, ¹H), 4.58–4.53 (m, ¹H), 3.70 (d, J = 8.0 Hz, 2H), 2.11–2.07 (m, ¹H), 1.99–1.97 (m, ¹H), 1.96 (s, ³H), 1.94–1.88 (m, ¹H), 1.80–1.02 (m, other alicyclic protons), 0.98 (s, ³H), 0.86–0.78 (m, ¹²H).

[0108] (3) Dissolve the product S-14 (20 mg, 0.0375 mmol) from the previous step in 10 mL of a mixed solvent of EtOH / H2O = 10:1, add potassium hydroxide (21.0 mg, 0.375 mmol), react at room temperature, monitor the reaction by TLC until complete, neutralize the reaction solution with 1N dilute hydrochloric acid to neutral, extract with ethyl acetate (3 × 20 mL), wash the combined organic layers with deionized water and saturated brine respectively, dry and concentrate with sodium sulfate, and separate by column chromatography to obtain compound C18 12 mg (0.0244 mmol), molar yield: 65.2%.

[0109] 1 ¹H NMR (400MHz, DMSO-d⁶) δ 8.07 (t, J = 6.0Hz, 1H), 3.70 (d, J = 5.8Hz, 2H), 3.29 (s, 1H), 2.48 (s, 1H), 2.02–1.85 (m, 2H), 1.73–1.01 (m, other alicyclic protons), 1.14 (s, 3H), 0.97 (s, 3H), 0.83–0.81 (m, 6H), 0.78 (s, 3H).

[0110] Example 5

[0111]

[0112] (1) Compound C2 (80 mg, 0.168 mmol), HOBT (22.7 mg, 0.168 mmol), DMAP (61.57 mg, 0.504 mmol), and EDCI (32.21 mg, 0.168 mmol) were dissolved in dichloromethane (10 mL), and D-alanine tert-butyl hydrochloride (45.80 mg, 0.252 mmol) was added. The reaction was carried out overnight at room temperature. After the reaction was monitored by TLC until it was complete, the reaction solution was evaporated to dryness and separated by column chromatography to obtain intermediate S-15 60 mg (0.099 mmol), with a molar yield of 59.2%. 1 ¹H NMR (400MHz, Chloroform-d) δ 7.00 (d, J = 7.3Hz, 1H), 4.72–4.63 (m, 1H), 4.50–4.43 (m, 1H), 2.62–2.56 (m, 1H), 2.42–2.32 (m, 1H), 2.14–2.09 (m, 1H), 2.02 (s, 3H), 1.92–1.03 (m, other alicyclic protons), 1.61 (s, 3H), 1.47 (s, 9H), 1.15 (s, 3H), 0.94 (d, J = 6.9Hz, 3H), 0.90 (s, 3H), 0.85 (d, J = 6.6Hz, 6H).

[0113] (2) Dissolve the product S-15 (60 mg, 0.099 mmol) from the previous step in dichloromethane (10 mL), add trifluoroacetic acid (0.73 mL, 99 mmol) dropwise, react at room temperature for 2 h, and monitor the reaction by TLC until it is complete. Then, evaporate the reverse chloroform to dryness and separate by column chromatography to obtain compound S-16 50 mg (0.091 mmol), molar yield: 92.3%. 1 ¹H NMR (400MHz, Chloroform-d) δ 7.51 (d, J = 6.3 Hz, 1H), 4.72–4.65 (m, 1H), 4.49–4.42 (m, 1H), 2.69–2.62 (m, 1H), 2.23 (d, J = 13.2 Hz, 1H), 2.11 (d, J = 12.8 Hz, 1H), 2.03 (s, 3H), 2.00–1.10 (m, other alicyclic protons), 1.52 (d, J = 7.2 Hz, 3H), 1.19 (s, 3H), 0.93 (d, J = 10.1 Hz, 6H), 0.86 (d, J = 6.6 Hz, 6H).

[0114] (3) Dissolve the product S-16 (50 mg, 0.091 mmol) from the previous step in 10 mL of a mixed solvent of EtOH / H2O = 10:1, add potassium hydroxide (50.9 mg, 0.91 mmol), react at room temperature, monitor the reaction by TLC until it is complete, neutralize the reaction solution with 1N dilute hydrochloric acid to neutral, extract with ethyl acetate (3 × 20 mL), wash the combined organic layers with deionized water and saturated brine respectively, dry and concentrate with sodium sulfate, and separate by column chromatography to obtain compound C19 40 mg (0.079 mmol), molar yield: 86.8%. 1 ¹H NMR (400MHz, Chloroform-d) δ 7.53–7.39 (m, 1H), 4.44 (s, 1H), 3.61–3.57 (m, 1H), 2.70–2.64 (m, 1H), 2.24 (s, 1H), 2.12–2.10 (m, 1H), 2.00–1.08 (m, other alicyclic protons), 1.52 (d, J = 8.0 Hz, 3H), 1.17 (d, J = 1.8 Hz, 3H), 0.93 (d, J = 6.9 Hz, 3H), 0.89 (s, 3H), 0.87–0.85 (m, 6H).

[0115] Example 6

[0116]

[0117] (1) Compound C2 (80 mg, 0.168 mmol), HOBT (22.7 mg, 0.168 mmol), DMAP (61.57 mg, 0.504 mmol), and EDCI (32.21 mg, 0.168 mmol) were dissolved in dichloromethane (10 mL), and L-alanine tert-butyl hydrochloride (45.80 mg, 0.252 mmol) was added. The reaction was carried out overnight at room temperature. After the reaction was monitored by TLC until it was complete, the reaction solution was evaporated to dryness and separated by column chromatography to obtain intermediate S-17 66.1 mg (0.109 mmol), molar yield: 64.9%. 1 ¹H NMR (400MHz, Chloroform-d) δ 6.85 (d, J = 7.7Hz, 1H), 4.72–4.64 (m, 1H), 4.51–4.44 (m, 1H), 2.60–2.53 (m, 1H), 2.42–2.39 (m, 1H), 2.18–2.05 (m, 2H), 2.02 (s, 3H), 1.90–1.02 (m, other alicyclic protons), 1.46 (s, 9H), 1.38 (d, J = 7.1Hz, 3H), 1.14 (s, 3H), 0.94 (d, J = 6.9Hz, 3H), 0.89 (s, 3H), 0.86–0.84 (m, 6H).

[0118] (2) Dissolve the product S-17 (60 mg, 0.099 mmol) from the previous step in dichloromethane (10 mL), add trifluoroacetic acid (0.73 mL, 99 mmol) dropwise, react at room temperature for 2 h, and monitor the reaction by TLC until it is complete. Then, evaporate the reverse chloroform to dryness and separate by column chromatography to obtain compound S-18 51.1 mg (0.093 mmol), molar yield: 93.4%. 1 ¹H NMR (400MHz, Chloroform-d) δ 7.48 (d, J = 6.4Hz, 1H), 4.71–4.63 (m, 1H), 4.52 (t, J = 7.0Hz, 1H), 2.65–2.58 (m, 1H), 2.31 (d, J = 7.4Hz, 1H), 2.25–2.08 (m, 2H), 2.02 (s, 3H), 1.96–1.00 (m, other alicyclic protons), 1.50 (d, J = 7.2Hz, 3H), 1.16 (s, 3H), 0.92 (d, J = 6.9Hz, 3H), 0.90 (s, 3H), 0.85 (d, J = 6.5Hz, 6H).

[0119] (3) Dissolve the product S-18 (50 mg, 0.091 mmol) from the previous step in 10 mL of a mixed solvent of EtOH / H2O = 10:1, add potassium hydroxide (50.9 mg, 0.91 mmol), react at room temperature, monitor the reaction by TLC until it is complete, neutralize the reaction solution with 1N dilute hydrochloric acid to neutral, extract with ethyl acetate (3 × 20 mL), wash the combined organic layers with deionized water and saturated brine respectively, dry and concentrate with sodium sulfate, and separate by column chromatography to obtain compound C20 34.9 mg (0.069 mmol), molar yield: 75.8%. 1 ¹H NMR (400MHz, Chloroform-d) δ 7.56 (s, 1H), 4.50 (s, 1H), 3.64–3.58 (m, 1H), 2.65 (t, J = 11.6 Hz, 1H), 2.26 (brs, 1H), 2.12 (d, J = 13.7 Hz, 1H), 2.00–1.08 (m, other alicyclic protons), 1.52 (d, J = 7.0 Hz, 3H), 1.18 (s, 3H), 0.93 (d, J = 6.9 Hz, 3H), 0.89 (s, 3H), 0.88–0.84 (m, 6H).

[0120] Example 7

[0121]

[0122] (1) Compound C2 (140 mg, 0.294 mmol), EEDQ (181.8 mg, 0.735 mmol), and taurine (73.6 mg, 0.588 mmol) were dissolved in dry DMF (10 mL), and triethylamine (0.41 mL, 2.94 mmol) was added dropwise. The reaction was carried out at 90 °C. After the reaction was completed by TLC monitoring, the reaction solution was cooled to room temperature, diluted with 50 mL of deionized water, and extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with deionized water and saturated brine, dried over anhydrous sodium sulfate and concentrated. The compound S-19 was separated by column chromatography to obtain 30 mg (0.051 mmol), with a molar yield of 17.5%. 1 ¹H NMR (400MHz, DMSO-d⁶) δ 7.67 (t, J = 5.5Hz, 1H), 4.57 (dq, J = 16.5, 6.0Hz, 1H), 3.30–3.22 (m, 2H), 3.09 (d, J = 7.5Hz, 2H), 2.47 (m, 2H), 1.97 (s, 3H), 1.94–0.99 (m, other alicyclic protons), 0.97 (s, 3H), 0.84 (d, J = 1.7Hz, 3H), 0.83 (d, J = 1.3Hz, 6H), 0.81 (s, 3H).

[0123] (2) Dissolve the product S-19 (30 mg, 0.051 mmol) from the previous step in 10 mL of a mixed solvent of EtOH / H2O = 10:1, add potassium hydroxide (28.6 mg, 0.51 mmol), react at room temperature, monitor the reaction by TLC until complete, neutralize the reaction solution with 1N dilute hydrochloric acid to neutral, extract with ethyl acetate (3 × 20 mL), wash the combined organic layers with deionized water and saturated brine respectively, dry and concentrate with sodium sulfate, and separate by column chromatography to obtain compound C21 10 mg (0.018 mmol), molar yield: 36.2%. 1 ¹H NMR (400MHz, Methanol-d⁴) δ 3.59–3.57 (m, 2H), 3.52–3.47 (m, 1H), 2.97–2.94 (m, 2H), 2.60–2.52 (m, 2H), 2.15 (s, 1H), 2.01–1.22 (m, other aliphatic ring protons), 1.10 (s, 3H), 0.94 (d, J = 8.0 Hz, 3H), 0.89 (d, J = 4.0 Hz, 6H), 0.87 (s, 3H).

[0124] Example 8

[0125]

[0126] (1) Dissolve intermediate S-7 (8 g, 18.69 mmol) in 70 mL of a mixed solvent of EtOH / H2O = 10:1, add potassium hydroxide (10.47 g, 1.87 mol), react at room temperature, monitor the reaction by TLC until complete, neutralize the reaction solution with 1N dilute hydrochloric acid to neutral, extract with ethyl acetate (3 × 100 mL), wash the combined organic layers with deionized water and saturated brine respectively, dry and concentrate with sodium sulfate, and separate by column chromatography to obtain intermediate S-20 6.75 g (17.49 mmol), molar yield: 93.6%. 1 ¹H NMR (400 MHz, Chloroform-d) δ 5.14 (s, 1H), 3.61 (m, 1H), 2.42–2.11 (m, 2H), 2.05–1.03 (m, other alicyclic protons), 0.91 (d, J = 8.2 Hz, 3H), 0.86–0.84 (m, 9H), 0.66 (d, J = 16.8 Hz, 3H).

[0127] (2) Dissolve the product S-20 (6.5 g, 16.84 mmol), imidazole (1.38 g, 20.21 mmol), and TBSCl (3.05 g, 20.21 mmol) from the previous step in dry DMF (30 mL). React at room temperature for 30 min. After the reaction is complete as monitored by TLC, dilute with 100 mL of deionized water and extract with ethyl acetate (3 × 100 mL). Wash the combined organic layers with deionized water and saturated brine, respectively. Dry and concentrate with anhydrous sodium sulfate. Separate by column chromatography to obtain intermediate S-217.9 g (15.80 mmol), molar yield: 93.8%. 1 ¹H NMR (400MHz, Chloroform-d) δ 5.14 (s, 1H), 3.58 (dd, J = 15.5, 9.0 Hz, 1H), 2.36 (d, J = 14.0 Hz, 1H), 2.22 (m, 2H), 2.04–0.98 (m, other alicyclic protons), 0.93 (d, J = 7.8 Hz, 3H), 0.82 (d, J = 9.4 Hz, 3H), 0.03 (s, 6H).

[0128] (3) Dissolve the product S-21 (7.5 g, 15 mmol) from the previous step in dichloromethane (50 mL), continuously pass O3 under an ice-water bath, and monitor the reaction by TLC until it is complete. Then replace the ozone in the system with argon, evaporate the reaction solution to dryness, and then obtain intermediate S-22 2.74 g (5.01 mmol) by column chromatography. The molar yield is 33.3%. 1 ¹H NMR (400MHz, Chloroform-d) δ 5.75 (d, J = 6.7Hz, 1H), 3.58–3.51 (m, 1H), 2.10–2.00 (m, 1H), 1.95–0.98 (m, other alicyclic protons), 0.94 (s, 3H), 0.91 (d, J = 6.9Hz, 3H), 0.88 (s, 9H), 0.86 (d, J = 6.6Hz, 6H), 0.77 (s, 3H), 0.05 (s, 6H).

[0129] (4) Dissolve the product S-22 (2.5 g, 4.56 mmol) and sodium borohydride (862.9 mg, 22.81 mmol) in tert-butanol (40 mL). Reflux the reaction mixture at 78 °C. After the reaction is complete as monitored by TLC, cool the reaction mixture to room temperature. Quench the reaction with deionized water (30 mL). Extract with ethyl acetate (3 × 100 mL). Wash the combined organic layers with deionized water and saturated brine, respectively. Dry and concentrate with anhydrous sodium sulfate. Separate by column chromatography to obtain intermediate S-23 1.52 g (2.84 mmol), molar yield: 62.3%.

[0130] 1 ¹H NMR (400MHz, Chloroform-d) δ 3.84–3.79 (m, 1H), 3.65–3.58 (m, 1H), 3.57–3.51 (m, 1H), 3.34 (s, 1H), 1.73–1.13 (m, other alicyclic protons), 0.96–0.94 (m, 3H), 0.88 (s, 9H), 0.87–0.85 (m, 9H), 0.77 (s, 3H), 0.05 (s, 6H).

[0131] (5) Under argon protection, the product S-23 (1.5 g, 2.80 mmol) and TEMPO (443.74 mg, 0.28 mmol) from the previous step were dissolved in dry dichloromethane (20 mL), and iodobenzene diacetate (1.08 g, 3.36 mmol) was added. The reaction was carried out overnight at room temperature. After the reaction was completed by TLC monitoring, the reaction solution was directly evaporated to dryness to give intermediate S-24 817.0 mg (1.53 mmol), molar yield: 54.6%.

[0132] (6) Take the product S-24 (100 mg, 0.187 mmol) and ethoxyformylmethylenetriphenylphosphine (326.19 mg, 0.936 mmol) from the previous step and dissolve them in toluene (20 mL). Transfer the reaction solution to 110 °C and react. After 72 h of reaction, TLC monitoring showed that the Rf values ​​of the starting material and the product were still the same. LCMS showed that the reaction was not complete. After cooling the reaction solution to room temperature, evaporate it to dryness and then separate it by column chromatography to obtain 90 mg of a mixture of intermediates S-25 and S-29. 1 ¹H NMR (400MHz, Chloroform-d) δ 7.13–7.05 (m, 1H), 5.85–5.77 (m, 1H), 4.23–4.12 (m, 2H), 3.58–3.49 (m, 1H), 3.28 (s, 1H), 2.41–2.34 (m, 1H), 2.26–2.17 (m, 1H), 1.80–1.00 (m, other alicyclic protons), 0.92–0.91 (m, 3H), 0.88 (s, 9H), 0.87–0.85 (m, 9H), 0.80 (s, 3H), 0.04 (s, 6H).

[0133] (7) Dissolve the mixture (90 mg) obtained in the previous step in EtOH (10 mL), replace with nitrogen, and then quickly add 10% Pd / C (10 mg by mass). Replace with nitrogen again, then with hydrogen, and stir at room temperature. When the reaction is complete as monitored by TLC, replace with nitrogen and filter out Pd / C. After the reaction solution is evaporated to dryness, perform column chromatography to obtain intermediate S-26 50 mg (0.0825 mmol). 1¹H NMR (400MHz, Chloroform-d) δ 4.15–4.10 (m, 2H), 3.57–3.49 (m, 1H), 3.30 (s, 1H), 2.32 (t, J = 7.0 Hz, 2H), 1.80–0.78 (m, other alicyclic protons), 0.96 (d, J = 7.1 Hz, 3H), 0.88 (s, 9H), 0.87–0.83 (m, 9H), 0.76 (s, 3H), 0.73 (s, 3H), 0.05 (s, 6H).

[0134] (8) Dissolve the product S-26 (50 mg, 0.0825 mmol) from the previous step in 10 mL of a mixed solvent of EtOH / H2O = 10:1, add potassium hydroxide (46.2 mg, 0.825 mol), react at room temperature, monitor the reaction by TLC until complete, neutralize the reaction solution with 1N dilute hydrochloric acid to neutral, extract with ethyl acetate (3 × 20 mL), wash the combined organic layers with deionized water and saturated brine respectively, dry and concentrate with sodium sulfate, and separate by column chromatography to obtain compound C24 35 mg (0.075 mmol), molar yield: 91.4%. 1 ¹H NMR (400MHz, Chloroform-d) δ 3.60 (dt, J = 11.1, 6.0 Hz, 1H), 3.33 (s, 1H), 2.38 (t, J = 7.1 Hz, 2H), 1.25 (s, 2H), 1.84–1.05 (m, other alicyclic protons), 0.97 (d, J = 7.2 Hz, 3H), 0.87 (d, J = 6.4 Hz, 6H), 0.77 (s, 3H), 0.74 (s, 3H).

[0135] Example 9

[0136]

[0137] (1) Dissolve intermediate S-24 (2 g, 3.75 mmol) in pre-dried pyridine (10 mL), add phosphorus oxychloride (0.7 mL, 7.5 mmol), and move the reaction to 110 °C. After the reaction is completed by TLC monitoring, the reaction solution is cooled to room temperature, and phosphorus oxychloride is quenched by adding 1 mL of water under ice-water bath conditions. After phosphorus oxychloride quenching, 20 mL of deionized water is added to the reaction solution, and the mixture is extracted with ethyl acetate (3 × 100 mL). The combined organic layers are washed with deionized water and saturated brine, respectively, dried and concentrated with anhydrous sodium sulfate, and separated by column chromatography to obtain intermediate S-27 1.85 g (3.59 mmol), with a molar yield of 95.7%.

[0138] 1¹H NMR (400MHz, Chloroform-d) δ 6.36 (dd, J = 6.3, 1.6 Hz, 1H), 4.59–4.56 (m, 1H), 3.60–3.47 (m, 2H), 1.82 (d, J = 8.2 Hz, 1H), 1.71–1.04 (m, other alicyclic protons), 0.96 (s, 3H), 0.91 (d, J = 6.8 Hz, 3H), 0.88 (s, 9H), 0.87 (d, J = 1.5 Hz, 3H), 0.85 (d, J = 1.6 Hz, 3H), 0.78 (s, 3H), 0.04 (s, 6H).

[0139] (2) Dissolve the product S-27 (1.85 g, 3.59 mmol) from the previous step in ethyl acetate (20 mL). Ozone was continuously introduced at -78 °C for 3 min. After the reaction was completed by TLC monitoring, the ozone in the reaction system was replaced with argon. Dimethyl sulfide was added to the reaction system, and after reacting for 5 min, the reaction solution was evaporated to dryness. The intermediate S-28 1.33 g (2.43 mmol) was obtained by column chromatography, with a molar yield of 67.7%.

[0140] 1 ¹H NMR (400MHz, Chloroform-d) δ 9.78 (d, J = 3.9 Hz, 1H), 8.07 (s, 1H), 4.92 (s, 1H), 3.57–3.49 (m, 1H), 2.14–2.12 (m, 1H), 1.93–1.80 (m, 2H), 1.73–0.92 (m, other alicyclic protons), 1.21 (s, 3H), 1.08 (d, J = 6.8 Hz, 3H), 0.87 (s, 9H), 0.86 (d, J = 1.2 Hz, 3H), 0.84 (d, J = 1.3 Hz, 3H), 0.78 (s, 3H), 0.04 (s, 6H).

[0141] (3) Dissolve the product S-28 (100 mg, 0.182 mmol) from the previous step in a mixed solvent (10 mL) of tert-butanol, tetrahydrofuran, and 2-methyl-2-butene in a volume ratio of 5:1:1.5. Dissolve sodium dihydrogen phosphate (199.2 mg, 1.82 mmol) and sodium chlorite (214.0 mg, 2.366 mmol) in 10 mL of water. Add the freshly prepared solution to the above reaction solution. React in an ice-water bath for 15 min, and then continue the reaction at room temperature. Monitor the reaction by TLC until complete. Extract with ethyl acetate (3 × 30 mL). Wash the combined organic layers with deionized water and saturated brine, respectively. Dry and concentrate with sodium sulfate. Separate by column chromatography to obtain compound S-29 84.5 mg (0.150 mmol), molar yield: 82.4%.

[0142] 1 ¹H NMR (400MHz, Chloroform-d) δ 8.12 (s, 1H), 5.03 (s, 1H), 3.56–5.50 (m, 1H), 2.33 (s, 1H), 1.90–1.17 (m, other alicyclic protons), 1.15 (s, 3H), 1.08 (d, J = 6.8 Hz, 3H), 0.87 (s, 9H), 0.85 (d, J = 6.6 Hz, 6H), 0.78 (s, 3H), 0.04 (s, 6H).

[0143] (4) Dissolve the product S-29 (84.5 mg, 0.150 mmol) from the previous step in dichloromethane, add trifluoroacetic acid (0.22 mL, 3.0 mmol) dropwise, react at room temperature, monitor the reaction by TLC until it is complete, evaporate the reaction solution to dryness, and separate by column chromatography to obtain compound C25 63.5 mg (0.141 mmol), molar yield: 94%.

[0144] 1 ¹H NMR (400MHz, Chloroform-d) δ 8.13 (s, 1H), 5.02 (s, 1H), 3.61–3.49 (m, 1H), 2.33 (s, 1H), 1.92–0.93 (m, other alicyclic protons), 1.16 (s, 3H), 1.08 (d, J = 4.0 Hz, 3H), 0.85 (d, J = 8.0 Hz, 6H), 0.79 (s, 3H).

[0145] (5) Dissolve the product C25 (60 mg, 0.13 mmol) from the previous step in 10 mL of a mixed solvent of EtOH / H2O = 10:1, add potassium hydroxide (74.5 mg, 1.33 mol), react at room temperature, monitor the reaction by TLC until it is complete, neutralize the reaction solution with saturated ammonium chloride solution to neutral, extract with ethyl acetate (3 × 100 mL), wash the combined organic layers with deionized water and saturated brine respectively, dry and concentrate with sodium sulfate, and separate by column chromatography to obtain compound C22 50 mg (0.118 mmol), molar yield: 91.1%.

[0146] 1 ¹H NMR (400MHz, Methanol-d⁴) δ 3.50 (d, J = 15.0 Hz, 2H), 2.46 (s, 1H), 1.83–0.93 (m, other alicyclic protons), 1.10 (d, J = 7.0 Hz, 3H), 1.07 (s, 3H), 0.88 (d, J = 6.5 Hz, 6H), 0.81 (s, 3H).

[0147] Example 10

[0148]

[0149] (1) Intermediate S-28 (100 mg, 0.182 mmol) and ethoxyformylmethylenetriphenylphosphine (635.7 mg, 1.82 mmol) were dissolved in toluene (20 mL). The reaction solution was transferred to 110 °C and reacted. After 96 h, TLC monitoring showed that the Rf values ​​of the product spot and the starting material spot were the same, and LCMS showed that the starting material had completely reacted. After the reaction solution was cooled to room temperature, rotary evaporation column chromatography was used to separate intermediate S-30 82 mg (0.132 mmol), with a molar yield of 72.8%.

[0150] 1 ¹H NMR (400MHz, Chloroform-d) δ 8.00 (s, 1H), 6.93–6.86 (m, 1H), 5.63 (d, J = 15.5 Hz, 1H), 4.21–4.15 (m, 2H), 3.56–3.49 (m, 1H), 2.02 (d, J = 11.3 Hz, 1H), 1.86–1.32 (m, other alicyclic protons), 1.29 (t, J = 7.2 Hz, 3H), 1.25 (s, 3H), 1.04 (s, 3H), 0.87 (s, 9H), 0.85 (d, J = 6.6 Hz, 6H), 0.77 (s, 3H), 0.03 (s, 6H).

[0151] (2) Dissolve the product S-30 (80 mg, 0.129 mmol) from the previous step in EtOH (10 mL). After changing the nitrogen atmosphere, quickly add 10% Pd / C (10 mg by mass), then change the nitrogen atmosphere again, followed by hydrogen atmosphere, and stir at room temperature. When the reaction is complete as monitored by TLC, change the nitrogen atmosphere again, filter out Pd / C, evaporate the reaction solution to dryness, and then perform column chromatography to obtain intermediate S-31 60 mg (0.119 mmol), molar yield: 75.2%.

[0152] 1 ¹H NMR (400MHz, Chloroform-d) δ 4.29–4.24 (m, 1H), 3.60–3.53 (m, 1H), 3.03 (d, J = 3.4 Hz, 1H), 2.66–2.60 (m, 1H), 2.39–2.34 (m, 1H), 1.82–0.96 (m, other alicyclic protons), 1.31–1.22 (m, 3H), 0.92 (d, J = 6.6 Hz, 3H), 0.86 (d, J = 6.6 Hz, 6H), 0.82 (s, 3H), 0.74 (s, 3H).

[0153] (3) Dissolve the product S-31 (50 mg, 0.099 mmol) from the previous step in 10 mL of a mixed solvent of EtOH / H2O = 10:1, add potassium hydroxide (45.1 mg, 0.80 mol), react at room temperature, monitor the reaction by TLC until it is complete, neutralize the reaction solution with 1N dilute hydrochloric acid to neutral, extract with ethyl acetate (3 × 20 mL), wash the combined organic layers with deionized water and saturated brine respectively, dry and concentrate with sodium sulfate, and separate by column chromatography to obtain compound C23 35 mg (0.078 mmol), molar yield: 78.8%.

[0154] 1 ¹H NMR (400MHz, Chloroform-d) δ 3.60 (s, 1H), 3.53 (s, 1H), 2.54–2.42 (m, 2H), 1.88–0.99 (m, other alicyclic protons), 0.96 (d, J = 6.8 Hz, 3H), 0.87 (d, J = 6.4 Hz, 6H), 0.77 (s, 3H), 0.72 (s, 3H).

[0155] Example 11 FXR Antagonism Test

[0156] 1. Experimental Objective

[0157] The antagonistic activity of the compound against FXR was detected using a reporter gene assay.

[0158] 2. Experimental Principle

[0159] Luciferase reporter gene assay is a reporter system that uses luciferin as a substrate to detect the activity of firefly luciferase. Luciferase catalyzes the oxidation of luciferin to oxyluciferin, during which bioluminescence is emitted. This bioluminescence can be measured using a fluorescence spectrometer. In this experiment, the agonist ligand binds to the ligand-binding domain (LBD) and promotes the binding of the DNA-binding domain (DBD) to DNA, initiating luciferase transcription. The addition of the substrate produces fluorescence, and the intensity of the fluorescence reflects the degree of receptor activation.

[0160] 3. Experimental Samples

[0161] Before the experiment, the compound was dissolved in DMSO to prepare a stock solution, which was then diluted with culture medium to the required concentration before use.

[0162] 4. Experimental Methods

[0163] Hek293T cells were grown to approximately 80% confluence, then digested and diluted to 2 x 10⁻⁶ cells. 5 A suspension of 100 μL / mL was seeded into each well of a white, opaque 96-well plate. Following the Lipofectamine 3000 kit instructions, serum-depleted opti-MEM medium was transferred 1:1 by volume into two separate centrifuge tubes. 20 μg / mL of plasmid and 2 μg / mL of liposomes were added to each tube, with the pCMV-Script-hFXR and pGL4.11-hSHP-Luciferase reporter plasmids premixed at a 1:1 ratio to a concentration of 20 μg / mL. The opti-MEM medium containing liposomes and the opti-MEM medium containing plasmids were then gently mixed and packaged at room temperature for 20 min. After packaging, 10 μL of the opti-MEM medium containing the plasmid-liposome mixture was added to each well of a 96-well plate and incubated at 37°C in a 5% CO2 incubator. After 24 hours, gently discard all the culture medium in the wells using a pipette, and replace it with medium containing 1% FBS for starvation treatment. After 6 hours, add the test compound in antagonistic mode, and then add GW4064 (final concentration 5 μM) after 1 hour. Continue incubation in an incubator for another 24 hours. Following the Firefly-Glo kit instructions, add the same volume of the test reagent as the culture medium to a 96-well plate, and allow it to fully lyse in the dark for 15 minutes. Then, read the fluorescence values ​​using a multi-label microplate reader.

[0164] Inhibition efficiency calculation: The ratio of the difference between 1 and the fluorescence intensity of the test compound minus the fluorescence intensity of the blank well to the difference between the fluorescence intensity of the activator model group minus the fluorescence intensity of the blank well is defined as the inhibition efficiency (Inhibition%) of the compound, as shown in the following formula:

[0165]

[0166] According to the above formula, the compound was tested at a concentration of 10 μM, with 3 replicates for each concentration (blank group: cells without drug treatment after plasmid co-transfection).

[0167]

[0168] The experimental results are shown in Table 1.

[0169] Table 1 Results of FXR antagonistic activity tests of compounds

[0170]

[0171] Note: Inhibition% is the rate of inhibition of the sample drug on the antagonistic activity of FXR, SD is the standard deviation, and LDH% represents cytotoxicity.

[0172] These compounds were able to compete with the FXR agonist GW4064 in HEK293T cells expressing the FXR ligand-binding domain and antagonize the FXR activation effect of GW4064.

[0173] As shown in Table 1, the compounds of this invention achieve an inhibition rate of 40% or even higher at 10 μM, and are FXR receptor antagonists. Compared with existing FXR receptor antagonists, these compounds have a novel structure containing a tricyclic structure, further enriching the variety of FXR receptor antagonists.

[0174] 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 of general formula (I), or a pharmaceutically acceptable salt thereof, (I) R1 is hydrogen; R2 is selected from: hydroxyl group, R d -C(=O)-O-; where R d The following groups may be substituted or unsubstituted: C3-C10 cycloalkyl, C6-C10 aryl, 5-7 heteroaryl; the substitution refers to the hydrogen on the group being replaced by one or more substituents selected from the group consisting of: halogen, C1-C6 alkyl, C1-C6 alkoxy; R3 is hydrogen; R4 is either absent or hydrogen; R5 is either =O or hydroxyl; R6 is selected from -(C1-C6 alkylene)C(=O)-R f -R g ,in, X is O; R c It is hydrogen; R f O is represented by 0; Rg is represented by hydrogen.

2. The compound according to claim 1, characterized in that, R2 is a hydroxyl group.

3. The compound according to claim 1, characterized in that, R2 is R d -C(=O)-O-; where R d The following groups may be substituted or unsubstituted: C3-C8 cycloalkyl, phenyl, 5-7 membered heteroaryl; The substitution refers to the replacement of hydrogen on a group by one or more substituents selected from the group consisting of halogens, C1-C6 alkyl groups, and C1-C6 alkoxy groups.

4. The compound according to claim 1, characterized in that, R4 indicates that the value does not exist, and R5 indicates that the value is 0.

5. The compound according to claim 1, characterized in that, R4 is hydrogen, and R5 is hydroxyl.

6. The compound according to claim 1, characterized in that, R6 is -(C1-C6 alkylene)COOH.

7. The compound according to claim 1, characterized in that, The compound is: 。 8. The method for preparing the compound according to claim 1, characterized in that, The compound has the structure shown in formula H3 and is prepared via the following route: The reaction of compound H1 with acyl chloride and acid anhydride yields compound H2; Compound H2 is debenzyl protecting group to give compound H3; R7 can be a substituted or unsubstituted group of the following: C3-C10 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl; The substitution refers to the replacement of hydrogen on a group by one or more substituents selected from the group consisting of halogens, C1-C6 alkyl groups, and C1-C6 alkoxy groups.

9. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the compound of any one of claims 1-7 or a pharmaceutically acceptable salt thereof; And pharmaceutically acceptable carriers.

10. Use of the compound according to any one of claims 1-7, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 9, characterized in that, (i) for the preparation of farnesoid derivative X receptor (FXR) antagonists; (ii) for the preparation of medicaments for the treatment of diseases associated with farnesoid derivative X receptors; or (iii) for the preparation of medicaments for the treatment of metabolic diseases.

11. The use as described in claim 10, characterized in that, The diseases associated with farnesoid derivative X receptors are selected from: hyperlipidemia, bile acid stasis, diabetes, obesity, non-alcoholic fatty liver disease, biliary cirrhosis, and hypercholesterolemia.

12. The use as described in claim 10, characterized in that, The metabolic diseases mentioned are selected from: hyperlipidemia, bile acid stasis, diabetes, obesity, non-alcoholic fatty liver disease, biliary cirrhosis, and hypercholesterolemia.

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