JP7450951B2 - Production of new FXR small molecule agonists and their use - Google Patents

Description

TECHNICAL FIELD The present invention is in the field of medicine and relates to the manufacture and use of non-steroidal compounds as FXR agonists. Specifically, small organic molecule compounds useful as FXR agonists and their enantiomers, diastereomers, tautomers, racemates, hydrates, solvates, prodrugs, or pharmaceutically acceptable salts thereof. and its use in the manufacture of medicaments for treating FXR-related diseases.

BACKGROUND ART Farnesoid X receptor is a member of the nuclear receptor superfamily, belongs to ligand-dependent nuclear transcription factors, and is mainly expressed in systems such as the liver, intestinal tract, kidney, and bile duct. FXR is activated by bile acids, which are endogenous ligands, and is an important part of bile acid metabolism and cholesterol metabolism, so it is also called the bile acid receptor. FXR is directly involved in the expression of over 300 genes, including physiological processes such as lipid metabolism, carbohydrate metabolism, inflammation, fibrosis, liver regeneration, and cell differentiation and proliferation. In its natural environment, its ligands include the primary bile acid chenodeoxycholic acid, the secondary bile acids lithocholic acid, deoxycholic acid, and the like. For example, FXR activated by its endogenous ligand bile acids plays an important role in the process of triglyceride (TG) metabolism, and FXR regulates the key enzymes, lipoproteins and corresponding receptors involved in TG metabolism. Through regulation, the TG content in the liver and circulating blood can be brought to a stable equilibrium. Therefore, there are already many FXR synthetic ligand molecules in the application field of metabolic diseases such as the liver.

FXR agonist molecules can be used to treat liver diseases such as primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC) and nonalcoholic fatty liver disease (NASH). It has already shown excellent clinical efficacy in treatment. Until now, obeticholic acid (OCA), the first FXR agonist molecule to be approved commercially, has shown significant improvements in many metabolic symptoms, such as lowering liver fat content and reducing inflammatory responses. and liver fibrosis. However, OCA also causes many clinical problems, such as itch induction, decrease in high-density lipoprotein cholesterol (HDLc), and increase in low-density lipoprotein cholesterol (LDLc). It has appeared, etc. Therefore, in terms of clinical demand, there is a strong desire for the emergence of new FXR agonist molecules with good clinical efficacy and low toxicity and side effects.
Additionally, one study demonstrated that FXR is closely related to tumor initiation and development. In many tumors, FXR plays the role of a tumor suppressor gene. For example, in hepatocellular carcinoma and rectal cancer, FXR is in a low expression state, and when FXR is activated, β-catenin activity is suppressed, and the progression of liver cancer or rectal cancer is significantly suppressed. Recent studies have shown that in cholangiocarcinoma, OCA, an FXR agonist, significantly inhibits the proliferation, migration, and clonogenesis of bile duct cells within the liver.

Additionally, FXR agonists are useful as new antiviral drug candidates, and one study showed that FXR ligands could be a potential therapeutic to inhibit the replication of a novel hepatitis B virus (HBV). be. FXR agonists can inhibit HBV surface antigen synthesis, inhibit HBV DNA and RNA replication, and most importantly, inhibit HBV cccDNA production. In hepatitis C virus (HCV), GW4064, an FXR agonist, can inhibit HCV invasion into liver tissue cells through indirect means. Therefore, FXR agonist molecules have great potential for the development of antiviral drugs.
As mentioned above, the field lacks novel FXR agonist molecules that are easy to prepare and have good inhibitory effects.

The aim of the present invention is to provide a new FXR agonist molecule that is easy to produce and has good inhibitory effects.
In a first aspect of the present invention, compounds represented by general formula I, or enantiomers, diastereomers, tautomers, racemates, hydrates, solvates, prodrugs thereof, or pharmaceutical Provide acceptable salt.
(however,
Ar is a substituted or unsubstituted C ₆ -C ₁₀ aryl group, a substituted or unsubstituted 5-9 membered heteroaromatic ring (including a single ring or a fused ring, and is selected from 1-3 oxygen, sulfur and nitrogen) containing heteroatoms).
A is a substituted or unsubstituted C ₆ -C ₁₀ aryl group, a substituted or unsubstituted 5-9 membered heteroaromatic ring (including a single ring or fused rings, and 1-3 atoms selected from oxygen, sulfur and nitrogen) containing heteroatoms).
R ¹ is a substituted or unsubstituted C ₁ -C ₆ alkyl group, a substituted or unsubstituted C ₃ -C ₆ cycloalkyl group, a substituted or unsubstituted 5-9 membered heterocycle (1-3 oxygen atoms, containing a heteroatom selected from sulfur and nitrogen).
X is selected from the group consisting of hydrogen or deuterium.
Here, the above substitution means that one or more hydrogen atoms in the group are each independently halogen, C ₁ -C ₆ haloalkyl group, C ₁ -C ₆ haloalkoxy group, C ₁ -C ₆ alkyl group, C It is substituted with a substituent selected from the group consisting of ₁ -C ₆ alkoxy group, C ₃ -C ₆ cycloalkyl group, C ₃ -C ₆ cycloalkyloxy group, cyano group or nitro group. )

In another preferred example, R ¹ is selected from the group consisting of a substituted or unsubstituted C ₁ -C ₆ alkyl group, a substituted or unsubstituted C ₃ -C ₆ cycloalkyl group, where: The above substitution means that one or more hydrogen atoms in the group are each independently halogen, C ₁ -C ₆ haloalkyl group, C ₁ -C ₆ haloalkoxy group, C ₁ -C ₆ alkyl group, C ₁ -C ₆ alkoxy group, C ₃ -C ₆ cycloalkyl group, C ₃ -C ₆ cycloalkyloxy group, cyano group or nitro group.

In another preferred example, the above Ar is selected from the group consisting of a substituted or unsubstituted C ₆ -C ₁₀ aryl group, a substituted or unsubstituted 5-9 membered heteroaromatic ring, and the above substituent is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, trifluoromethyl, or trifluoromethoxy.

In another preferred example, said A is selected from the group consisting of a substituted or unsubstituted C ₆ -C ₁₀ aryl group, a substituted or unsubstituted 5-9 membered heteroaromatic ring; Substituents on aryl or heteroaryl groups include hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, trifluoromethyl, or trifluoromethoxy. selected from the group consisting of
In another preferred example, Ar is a substituted or unsubstituted phenyl group, a substituted or unsubstituted 5-7 membered heteroaromatic ring (including a single ring or fused rings, containing 1-3 oxygen, sulfur and nitrogen containing a heteroatom selected from the group consisting of

In another preferred example, A is a substituted or unsubstituted phenyl group, a substituted or unsubstituted 5-7 membered heteroaromatic ring (including a single ring or fused rings, containing 1-3 oxygen, sulfur and nitrogen containing a heteroatom selected from the group consisting of
In another preferred example, A is substituted or unsubstituted benzothiazole.
In another preferred example, Ar or A is each independently a substituted or unsubstituted benzene ring, pyridine ring, pyrimidine ring, pyridazine ring, pyrimidine ring, pyridazine ring, furan ring, thiophene ring, pyrrole ring, thiazole ring, or imidazole ring.

In another preferred example, R ¹ is selected from the group consisting of a substituted or unsubstituted C _1-4 alkyl group and a substituted or unsubstituted cyclopropyl group.
In another preferred example, the above Ar is a substituted or unsubstituted benzene ring.
In another preferred example, said Ar is selected from the group consisting of 2,5-dichlorophenyl, 2-methylphenyl, 2-trifluoromethylphenyl, 2-trifluoromethoxyphenyl.
In another preferred example, R ¹ is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, cyclopropyl, cyclobutyl or cyclopentyl. .

In another preferred example, the compound of formula (I) has a structure represented by the following formula.
In another preferred example, the compound of formula (I) has a structure represented by the following formula.

In another preferred example, said compound is selected from the group below.
A second aspect of the invention provides a method for producing a compound according to the first aspect of the invention, comprising the steps of producing a compound of formula I by the method described in Scheme 1 or Scheme 2 below. provide:

(a) Using a substituted benzaldehyde, a compound represented by the general formula II, as a starting material, it is reacted with hydroxyamine hydrochloride under the action of a base to obtain an intermediate, which is then salified with N-chlorosuccinimide (NCS) to obtain a general becomes a compound represented by formula III;
(b) then reacting the compound of general formula III with the corresponding 3-oxopropionic ester under the action of a base to obtain a compound of general formula IV;
(c) reducing the ester in the compound of general formula IV to the corresponding alcohol, i.e. the compound of general formula V, under the action of a deuterated reducing agent;
(d) brominating a compound represented by general formula V with a bromine reagent to produce a compound represented by VI;
(e) reacting a compound represented by general formula VI and a compound represented by VII under the action of a base to form a compound represented by general formula VIII;
(f) reacting a compound of general formula VIII with hydroxyamine hydrochloride under the action of a base to produce a compound of general formula IX;
(g) reacting a compound of general formula IX under the action of phosgene, triphosgene or carbonyldiimidazole to produce a compound of general formula I;
(However, X is deuterium, and the definitions of R ¹ , Ar, and A are as described in the first aspect of the present invention.)

(a) reducing the ester in the compound of general formula IV to the corresponding alcohol, i.e. the compound of general formula X, under the action of a reducing agent;
(b) oxidizing the compound of general formula X to the corresponding aldehyde, i.e. the compound of general formula XI, under the action of an oxidizing agent;
(c) reducing the ester in the compound of general formula XI to a compound of general formula V under the action of a deuterated reducing agent;
(d) brominating a compound represented by general formula V with a bromine reagent to produce a compound represented by VI;
(e) reacting a compound represented by general formula VI and a compound represented by VII under the action of a base to form a compound represented by general formula VIII;
(f) reacting a compound of general formula VIII with hydroxyamine hydrochloride under the action of a base to produce a compound of general formula IX;
(g) Reacting a compound of general formula IX under the action of phosgene, triphosgene or carbonyldiimidazole to produce a compound of general formula I.
(In each formula, X is hydrogen, and the definitions of R ¹ , Ar, and A are as described in the first aspect of the present invention.)

In another preferred example, the compound of general formula VII is prepared by the following steps:
(k) A compound represented by general formula XII and a compound represented by general formula XIII are reacted under the action of a base to produce a compound represented by general formula VII.
(In each formula, the definition of A is as described in the first aspect of the present invention.)

In another preferred example, if optical isomers exist in the product, they are prepared with raw materials of corresponding optical configuration.
In a third aspect of the present invention, a compound represented by general formula I according to the first aspect of the present invention, or an enantiomer, diastereomer, tautomer, racemate, hydrate, or solvate thereof, is provided. A pharmaceutical composition comprising a drug, a prodrug, a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier is provided.

A fourth aspect of the present invention provides a compound represented by the general formula I according to the first aspect of the present invention, or an enantiomer, diastereomer, tautomer, racemate, hydrate, or solvate thereof. The present invention provides the use of a compound, a prodrug, or a pharmaceutically acceptable salt thereof for the manufacture of a pharmaceutical composition for treating a disease or disease related to the activity or expression level of FXR.
In another preferred example, said FXR-related disease is selected from the group consisting of diseases associated with processes of bile acid metabolism, carbohydrate metabolism, lipid metabolism, inflammation, and/or liver fibrosis.

In another preferred example, the FXR-related disease is non-alcoholic fatty liver (NASH), primary biliary cirrhosis (PBC), primary biliary cirrhosis (PSC), gallstones, non-alcoholic cirrhosis, hepatitis B. (HBV), hepatitis C (HCV), liver fibrosis, cholestatic liver disease, hyperlipidemia, hypercholesterolemia, or diabetes.
In another preferred example, the pharmaceutical composition described above is used as an FXR agonist.
In another preferred example, said pharmaceutical composition is used for lowering the levels of ALP, ALT, AST, TBA in serum.
In another preferred example, the pharmaceutical composition described above is used for lowering the content of hydroxyproline in liver tissue.

In another preferred example, said pharmaceutical composition is used to down-regulate the expression of α-SMA and Col1α1 mRNA in liver tissue.
In another preferred embodiment, the pharmaceutical composition is used to inhibit the synthesis of HBV surface antigen, inhibit the replication of HBV DNA and RNA, and inhibit the production of HBV cccDNA.
In another preferred example, said pharmaceutical composition is used for reducing the content of collagen in the liver.

In another preferred example, said pharmaceutical composition is prepared by mixing a compound of formula I with medicinal adjuvants (e.g. excipients, diluents, etc.) and forming tablets, capsules, granules or tablets for oral administration. It is manufactured by a method of preparing syrup, etc.
Of course, it is understood that within the scope of the present invention, each of the above-mentioned technical features of the present invention and each of the specifically described technical features below (for example, in the embodiments) can be combined with each other to constitute a new or preferred technical solution. be done. Due to space limitations, we will not explain each part here.

Specific Embodiments After extensive and deep research, the inventor of the present application has researched and developed a non-steroidal compound useful as an FXR agonist, which has the ability to act on FXR at both the molecular and cellular levels. However, in studies, the present compound lowered the levels of ALP, ALT, AST, and TBA in serum, lowered the content of hydroxyproline in liver tissue, and decreased the expression of α-SMA and Col1α1 mRNA in liver tissue. It was shown that it can down-regulate and reduce the content of collagen in the liver, suppress the synthesis of HBV surface antigen, suppress the replication of HBV DNA and RNA, and suppress the production of HBV cccDNA. . The compounds of the present invention have the advantages of high FXR agonist activity, easy synthesis, and easy availability of raw materials, so they can be used in the production of drugs for treating FXR-related diseases. Based on this, the present invention was completed.

Terminology In the present invention, unless otherwise specified, the terms used have their common meanings known to those skilled in the art.
In the present invention, the halogen is F, Cl, Br or I.
In the present invention, the term "C1-C6" means having 1, 2, 3, 4, 5 or 6 carbon atoms, and "C3-C6" means having 3, 4, 5 or 6 carbon atoms. This analogy can be made by having atoms.
In the present invention, the term "alkyl group" refers to a saturated linear or branched hydrocarbon moiety; for example, the term "C1-C6 alkyl group" refers to a linear or branched alkyl group having 1 to 6 carbon atoms. The groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, t-butyl, pentyl and hexyl groups, preferably ethyl, propyl, isopropyl, It includes, but is not limited to, butyl, isobutyl, sec-butyl and t-butyl.

In the present invention, the term "alkoxy group" represents an -O-(C1-C6 alkyl) group. For example, the term "C1-C6 alkoxy" refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy or butoxy. including but not limited to.
In the present invention, the term "cycloalkyl group" refers to a saturated cyclic hydrocarbon group, for example, the term "C3-C6 cycloalkyl group" refers to a cyclic alkyl group having 3 to 6 carbon atoms in the ring. This includes, but is not limited to, a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, and the like.

In the present invention, the term "cycloalkyloxy group" refers to a cycloalkyl-O- group, where the cycloalkyl group is as described above.
In the present invention, the term "aryl group" refers to a portion of a hydrocarbon group containing one or more aromatic rings. Examples of aryl groups include, but are not limited to, phenyl (Ph), naphthyl, pyrenyl, fluorenyl, anthryl and phenanthryl groups.
In the present invention, the term "heteroaryl group" refers to a moiety having one or more aromatic rings containing at least one heteroatom (eg N, O or S). Examples of heteroaryl groups include furyl, pyrrolyl, thienyl, oxazolyl, imidazolyl, thiazolyl, pyridyl, pyrimidinyl, quinazolinyl, quinolinyl, isoquinolinyl, indolyl, and the like.

Unless otherwise stated, the alkyl, alkoxy, cycloalkyl, cycloalkyloxy, aryl, and heteroaryl groups described herein are substituted and unsubstituted groups. Possible substituents on alkyl, alkoxy, cycloalkyl, cycloalkyloxy, aryl and heteroaryl groups include hydroxy, amino, nitro, nitrile, halogen, C ₁ -C ₆ alkyl, _C2 - _C10 alkenyl group, _C2 - _C10 alkynyl group, C3 _{- C20} cycloalkyl group, _C3 - _C20 cycloalkenyl group, _C1 - _C20 heterocycloalkyl group, _C1 - _C20 hetero Cycloalkenyl group, C1- _C6 alkoxy group, aryl group, heteroaryl group, heteroaloxyl group, _C1 - _C10 alkylamino group, _C1 - _C20 dialkylamino group, arylamino group, _diarylamino group, C ₁ -C ₁₀ alkylaminosulfonyl group, arylaminosulfonyl group, C ₁ -C ₁₀ alkylimino group, C ₁ -C ₁₀ alkylsulfonylimino group, arylsulfonylimino group, mercapto group, C ₁ -C ₁₀ alkylthio group, C ₁ -C ₁₀ alkylsulfonyl group, arylsulfonyl group, acylamino group, aminoacyl group, aminothioacyl group, guanidyl group, urea group, cyano group, acyl group, thioacyl group, acyloxy group, carboxy group and carboxylic acid ester group including but not limited to. On the other hand, cycloalkyl groups, heterocycloalkyl groups, heterocycloalkenyl groups, aryl groups and heteroaryl groups may also be condensed with each other.

In the present invention, said substitution is monosubstitution or polysubstitution, and said polysubstitution is disubstitution, trisubstitution, tetrasubstitution, or pentasubstitution. The above-mentioned disubstitution means having two substituents, which is analogous to this.
The pharmaceutically acceptable salts according to the present invention may be salts consisting of an anion and a positively charged group in a compound of formula I. Suitable anions include chloride, bromide, iodine, sulfate, nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate, acetate, malate, toluenesulfonate, These are tartrate ion, fumarate ion, glutamate ion, glucuronate ion, lactate ion, glutarate ion, or maleate ion. Similarly, a salt consisting of a cation and a negatively charged group in a compound of formula I may be used. Suitable cations include sodium, potassium, magnesium, calcium and ammonium ions, such as tetramethylammonium ion.

In another preferred example, "pharmaceutically acceptable salts" refers to salts of formula I with hydrofluoric acid, hydrochloric acid, hydrobromic acid, phosphoric acid, acetic acid, oxalic acid, sulfuric acid, nitric acid, methanesulfonic acid, etc. Acids, aminosulfonic acid, salicylic acid, trifluoromethanesulfonic acid, naphthalenesulfonic acid, maleic acid, citric acid, acetic acid, lactic acid, tartaric acid, succinic acid, oxalic acid, pyruvic acid, malic acid, glutamic acid, p-toluenesulfonic acid, naphthalene Sulfonic acid, naphthalenedisulfonic acid, malonic acid, fumaric acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid , 2-acetyloxybenzoic acid and hydroxyethanesulfonic acid, or sodium, potassium, calcium, aluminum or ammonium salts of a compound of formula I and an inorganic base, or the general formula Methylamine salt, ethylamine salt, or ethanolamine salt consisting of I compound and an organic base.
In another preferred example, in the above compounds, any one of ring A, Ar, X and R ¹ is each the corresponding group in the specific compounds described in the Examples.
The compounds of the invention have asymmetric centers, chiral axes and chiral faces, and may exist in the form of racemates, R-isomers or S-isomers. A person skilled in the art can resolve the racemate to obtain the R-isomer and/or the S-isomer by conventional technical means.

Manufacturing method The method for manufacturing the compound represented by the general formula I of the present invention is as shown in the synthesis scheme below.
(a) Using a substituted benzaldehyde, a compound represented by the general formula II, as a starting material, it is reacted with hydroxyamine hydrochloride under the action of a base to obtain an intermediate, which is then salified with N-chlorosuccinimide (NCS) to obtain a general becomes a compound represented by formula III;
(b) then reacting the compound of general formula III with the corresponding 3-oxopropionic ester under the action of a base to obtain a compound of general formula IV;
(c) reducing the ester in the compound of general formula IV to the corresponding alcohol, i.e. the compound of general formula V, under the action of a deuterated reducing agent;
(d) brominating a compound represented by general formula V with a bromine reagent to produce a compound represented by VI;
(e) reacting a compound represented by general formula VI and a compound represented by VII under the action of a base to form a compound represented by general formula VIII;
(f) reacting a compound of general formula VIII with hydroxyamine hydrochloride under the action of a base to produce a compound of general formula IX;
(g) Reacting a compound of general formula IX under the action of phosgene, triphosgene or carbonyldiimidazole to produce a compound of general formula I.
(However, X is deuterium, and the definitions of R ¹ , Ar, and A are as described in claim 1.)

(h) reducing the ester in the compound of general formula IV to the corresponding alcohol, i.e. the compound of general formula X, under the action of a reducing agent;
(i) oxidizing the compound of general formula X to the corresponding aldehyde, i.e. the compound of general formula XI, under the action of an oxidizing agent;
(j) reducing the ester in the compound of general formula XI to the compound of general formula V under the action of a deuterated reducing agent;
(d) brominating a compound represented by general formula V with a bromine reagent to produce a compound represented by VI;
(e) reacting a compound represented by general formula VI and a compound represented by VII under the action of a base to form a compound represented by general formula VIII;
(f) reacting a compound of general formula VIII with hydroxyamine hydrochloride under the action of a base to produce a compound of general formula IX;
(g) Reacting a compound of general formula IX under the action of phosgene, triphosgene or carbonyldiimidazole to produce a compound of general formula I.
(In each formula, X is hydrogen, and the definitions of R ¹ , Ar, and A are as described in the first aspect of the present invention.)

In another preferred example, the compound of general formula VII is prepared by the following steps:
(k) A compound represented by general formula XII and a compound represented by general formula XIII are reacted under the action of a base to produce a compound represented by general formula VII.
(In each formula, the definition of A is as described in the first aspect of the present invention.)

PHARMACEUTICAL COMPOSITIONS AND THERAPEUTIC USE THEREOF The compounds provided by the present invention may be used alone or they may be mixed with medicinal adjuvants (e.g. excipients, diluents, etc.) for oral administration. It may be prepared into tablets, capsules, granules, syrups, etc. The pharmaceutical composition can be manufactured by conventional methods in pharmaceutical science. The pharmaceutical compositions of the present invention contain an active ingredient within a safe and effective amount and a pharmaceutically acceptable carrier.
An "active ingredient" according to the present invention refers to a compound of formula I according to the present invention.
The "active ingredients" and pharmaceutical compositions according to the present invention are used for the manufacture of medicaments for treating FXR-related diseases. The "active ingredients" and pharmaceutical compositions described in this invention are useful as FXR agonists. In another preferred example, the active ingredients described above can be used for the manufacture of a medicament for preventing and/or treating diseases modulated by FXR agonists.

A "safe and effective amount" refers to an amount of active ingredient that is sufficient to significantly improve a medical condition and does not cause serious side effects. Typically, pharmaceutical compositions contain 1 to 2000 mg, preferably 10 to 200 mg of active ingredient per unit dosage. Preferably, said "unit dosage" is one tablet.
"Pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid filler or gel substances that are applicable to humans and must have sufficient purity and sufficiently low toxicity. "Compatible" means that the components in the composition can be blended with the active ingredients of the present invention and with each other without significantly reducing the effectiveness of the active ingredients. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (e.g. sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g. stearic acid, magnesium stearate), sulfuric acid. Calcium, vegetable oils (e.g. soybean oil, sesame oil, peanut oil, olive oil, etc.), polyhydric alcohols (e.g. propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g. Twin®), wetting agents (e.g. dodecyl sulfate). (sodium), colorants, seasonings, stabilizers, antioxidants, preservatives, pyrogen-free distilled water, etc.

The mode of application of the active ingredient or pharmaceutical composition of the present invention is not particularly limited, but typical modes of application include oral administration, intratumor, rectal, extra-gastrointestinal (intravenous, intramuscular, or subcutaneous), etc. , but not limited to.
Solid dosage forms used for oral administration include capsules, tablets, pills, powders and granules.
Liquid dosage forms used for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. Besides the active ingredient, the liquid dosage form contains inert diluents commonly used in the field, such as water or other solvents, compatibilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, It may also include 1,3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn oil, olive oil, castor oil or sesame oil or mixtures of these substances. Besides these inert diluents, the compositions can also contain auxiliary agents, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.

In addition to the active ingredients, suspensions may contain suspending agents, such as ethoxylated isooctadecanol, polyoxyethylene sorbitol or sorbitan esters, microcrystalline cellulose, methoxyaluminum or agar or mixtures of these substances. good.
Compositions for parenteral injection include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions and emulsions, and sterile preparations for reconstitution into sterile injectable solutions or dispersions. Contains powder. Suitable aqueous or non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyhydric alcohols and appropriate mixtures thereof.
The compounds of the invention may be administered alone or in combination with other therapeutic agents (eg, hypolipidemic agents).

When using a pharmaceutical composition, a safe and effective amount of a compound of the present invention is used in a mammal (e.g., a human) in need of treatment, and the dosage used is a dosage that is known to be pharmaceutically effective. For a human weighing 60 kg, the daily dose is usually 1-2000 mg, preferably 20-500 mg. Of course, the specific dosage should further take into account factors such as the mode of administration and the health status of the patient, all of which are within the skill of a skilled physician.

The present invention will be further explained below with reference to specific examples. It is understood that these examples are only used to illustrate the invention and are not intended to limit the scope of the invention. Experimental methods for which specific conditions are not given in the examples below are generally described, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Publishers, 1989). or in accordance with the manufacturer's recommended conditions. Unless otherwise stated, percentages and parts are by weight.
Unless otherwise defined, all technical and scientific terms used herein have meanings that are familiar to those of ordinary skill in the art. Additionally, any methods and materials similar or equivalent to those described can be used in the method of the present invention. The preferred implementation methods and materials described herein are for illustration only.

The equipment and main experimental materials used are as follows.
The reagents and anhydrous solvents used were purchased from China Commercial Corporation, and all were used as received unless otherwise stated. For ¹ H and ¹³ C NMR, Bruker AM-400 and Varian Mercury plus-400 nuclear magnetic resonance instruments were used, for mass spectrometry, an Agilent 6230 mass spectrometer was used, and for 200-300 mesh column chromatography on silica gel (Qingdao HSGF254 TLC plate (Yantai Chemical Industry Research Institute).
Compounds of the present invention were prepared using appropriate starting materials according to either Scheme 1 or Scheme 2 below.

Synthesis of Example Intermediate VII-1:
Endo-8-azabicyclo[3.2.1]octan-3-ol XII (10 g, 78.7 mmol) and p-fluorobenzonitrile XIII-1 (78.7 mmol) were dissolved in N,N-dimethylformamide (150 mL), Potassium carbonate (197 mmol) was added in portions at room temperature, and the mixture was reacted at 80°C overnight. The reaction was diluted with ethyl acetate (500 mL), washed with water, and the aqueous phase was extracted with ethyl acetate (3 x 300 mL). The organic phases were combined, washed with saturated brine, and concentrated. Intermediate VII-1 (11 g, yield 61%) was obtained by column chromatography. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.51 - 7.48 (m, 2H), 6.82 - 6.79 (m, 2H), 4.62 (s, 1H), 4.27 (d, J = 5.2 Hz, 2H), 3.78 (s, 1H), 2.28 (d, J = 6.8 Hz, 2H), 1.91 - 1.85 (m, 4H), 1.57 (d, J = 14.0 Hz, 2H). MS(ESI, m/z): 229 [M+H] ⁺ .

Synthesis of Example Compound 1:

Add potassium carbonate aqueous solution (3N, 182 mmol) drop by drop to a stirring solution of hydroxyamine hydrochloride (182 mmol) in ethanol (100 mL) at 0°C, and add 2,6-dichlorobenzaldehyde II-1 (20 g, 114 mmol) in 100 mL of ethanol, the solution was added to the above reaction solution, the temperature was raised to 90°C, and the mixture was reacted for 2 hours. Once the reaction was cooled to room temperature, it was concentrated to a solid. A water/ethanol (1000 mL/100 mL) solution was added and stirred to crush the solid, filtered, and vacuum dried at 50° C. overnight to obtain a compound intermediate (18.4 g). Dissolve the intermediate in N,N-dimethylformamide (50 mL), add dropwise to a solution of N-chlorosuccinimide (97 mmol) in N,N-dimethylformamide (100 mL) at 0 °C, and stir overnight. did. The reaction solution was poured into ice water at 0°C, extracted with methyl-t-butyl ether (200 mL each, three times in total), and the organic phase was washed with saturated brine and concentrated to obtain a crude product. Add n-hexane (600 mL) to the flask containing the crude product, stir magnetically, filter, and dry the solid in vacuo (30oC) to obtain intermediate III-1 (18.3 g, 73% yield). Ta. ^1H NMR (400 MHz, _CDCl3 ) δ 7.43 - 7.39 (m, 2H), 7.39 - 7.33 (m, 1H).

Triethylamine (8.2 g) was added to methyl 3-cyclopropyl-3-oxopropionate (82 mmol) and stirred for 30 minutes. Then, cool to 10°C, add a solution of intermediate III-1 (18.3 g, 82 mmol) in absolute ethanol (80 mL) drop by drop (so that the internal temperature is below 30°C), and let it cool at room temperature. The reaction was allowed to proceed overnight. The reaction was diluted with ethyl acetate (100 mL), washed with water, and the aqueous phase was extracted with ethyl acetate (3 x 100 mL). The organic phases were combined, washed with saturated brine, and concentrated. The concentrate was stirred with 100 mL of ethyl ether and the solvent was removed in vacuo to give a solid product, Compound IV-1 (21.6 g, 84% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.43 - 7.39 (m, 2H), 7.39 - 7.33 (m, 1H), 3.72 (s, 3H), 2.21 - 2.09 (m, 1H), 1.35 - 1.28 (m , 2H), 1.25 - 1.18 (m, 2H); MS(ESI, m/z): 312[M+H] ⁺ .

Lithium aluminum deuteride (7.3 g) was placed in tetrahydrofuran (200 mL), cooled to 0°C, and a solution of compound IV-1 (21.6 g, 69.2 mmol) in tetrahydrofuran (50 mL) was added dropwise (internally). The reaction solution was stirred at room temperature for 2 h so that the temperature was below 5°C. Quench the reaction by adding ice water (9 mL) at 0°C, then add one drop each of 15% aqueous sodium hydroxide solution (9 mL) and ice water (27 mL), and then add anhydrous magnesium sulfate (100 g). The mixture was stirred at room temperature for 0.5 h, filtered, concentrated, and subjected to column chromatography to obtain Intermediate V-1 (19 g, yield 96%). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.72 - 7.59 (m, 2H), 7.58 - 7.50 (m, 1H), 4.90 (s, 1H), 2.38 - 2.26 (m, 1H), 1.16 - 1.03 (m, 4H). MS(ESI, m/z): 286[M+H] ⁺ .

Compound V-1 (19 g, 66.4 mmol) was dissolved in dichloromethane (200 mL), cooled to 0°C, phosphorus tribromide (66.4 mmol) was slowly added dropwise to the solution, and the reaction solution was stirred at room temperature for 2 h. did. The solvent was removed from the reaction solution to obtain an oil, which was then added with ethyl acetate (100 mL), the pH value of the reaction solution was adjusted to neutrality with saturated aqueous sodium bicarbonate solution, washed with water, and then diluted with acetic acid. The aqueous phase was extracted with ethyl (3 times, 100 mL each). The organic phases were mixed, washed with saturated brine, and then concentrated. Intermediate VI-1 (20.2 g, yield 87%) was obtained by column chromatography. ¹ H NMR (400 MHz, DMSO- _d6 ) δ 7.72 - 7.65 (m, 2H), 7.64 - 7.56 (m, 1H), 2.48 - 2.39 (m, 1H), 1.26 - 1.10 (m, 4H). MS (ESI, m/z): 348[M+H] ⁺ .

Potassium t-butoxide (21.4 mmol) was added to a solution of compound VII-1 (1.96 g, 8.6 mmol) in anhydrous tetrahydrofuran (150 mL) at 0°C, and after stirring for 15 minutes, compound VII-1 (8.6 mmol) was added dropwise. mmol) in anhydrous tetrahydrofuran (50 mL), and the reaction mixture was stirred at room temperature for 4 h. Water (200 mL) was added to the reaction solution, extracted with ethyl acetate (200 mL each, 3 times in total), the organic phase was washed with saturated brine, concentrated, and the intermediate VIII-1 (2.1 g, yield 49%). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.73-7.63 (m, 2H), 7.64 - 7.55 (m, 3H), 6.83 (d, J = 8.4 Hz, 2H), 4.16 (s, 2H), 3.37 (s, 1H), 2.38 - 2.31 (m, 1H), 1.83 - 1.74 (m, 6H), 1.56 - 1.49 (m, 2H), 1.16 - 1.06 (m, 4H). MS(ESI, m/z ):496[M+H] ⁺ .

Intermediate VIII-1 (2.1 g, 4.2 mmol), hydroxyamine hydrochloride (8.4 mmol), and absolute ethanol (80 mL) were placed in a round bottom flask and stirred, triethylamine (8.4 mmol) was slowly added dropwise, and the mixture was heated to 80 °C. The mixture was heated and reacted overnight. Cool to room temperature, remove the solvent, dissolve in dichloromethane (150 mL), wash with water and saturated brine, concentrate the organic phase, and obtain Intermediate IX-1 (1.1 g, yield) by silica gel column chromatography. 50%). ¹ HNMR (400 MHz, DMSO-d ₆ ) δ 7.73 - 7.63 (m, 2H), 7.64 - 7.55 (m, 3H), 6.83 (d, J = 8.4 Hz, 2H), 4.67 (s, 2H), 3.43 - 3.35 (m, 1H), 2.39 - 2.32 (m, 1H), 1.89 - 1.78 (m, 6H), 1.57 - 1.48 (m, 2H), 1.16 - 1.06 (m, 4H). MS(ESI, m/ z):529[M+H] ⁺ .

Intermediate IX-1 (1.1 g, 2.1 mmol), N,N'-carbonyldiimidazole (3.1 mmol), and 1,4-dioxane (100 mL) were placed in a round bottom flask, and then 1,8-diazabicyclo[ 5.4.0] undecene-7 (3.1 mmol) was added, heated to 100°C, and reacted for 8 hours. The reaction station was cooled to room temperature, diluted with water (100 mL), adjusted to pH approximately 3 with 1M aqueous hydrochloric acid solution, and extracted with ethyl acetate (100 mL each, three times in total). The organic phases were combined, washed with saturated brine, and concentrated, and the resulting crude product was further subjected to silica gel column chromatography to obtain the final product, Compound 1 (158 mg, yield 13%). ¹ HNMR (400 MHz, DMSO-d ₆ ) δ 7.65 - 7.63 (m, 2H), 7.59 - 7.55 (m, 3H), 6.83 (d, J = 8.4 Hz, 2H), 4.17 (s, 2H), 3.38 (s, 1H), 2.37 - 2.30 (m, 1H), 1.80 - 1.72 (m, 6H), 1.51 (d, J = 14.4 Hz, 2H), 1.16 - 1.06 (m, 4H). MS(ESI, m /z):555[M+H] ⁺ .

Example 2:
Example 2 was produced from Intermediate VII-2 according to Scheme 1, and the synthesis scheme is as follows.

After synthesizing compound VII-2 by a synthetic method of synthesizing intermediate VII-1 from raw material XIII-2, compound 2 was obtained according to scheme 1, where:
White solid compound VIII-2: yield 77%, ¹ HNMR (400 MHz, DMSO-d ₆ ) δ8.21 (s, 1H), 7.70 - 7.62 (m, 3H), 7.60 - 7.54 (m, 1H) , 7.16 (d, J = 8.4 Hz, 1H), 4.29 (s, 2H), 3.43 - 3.35 (m, 1H), 2.37 - 2.29 (m, 1H), 1.85-1.65 (m, 6H), 1.63 - 1.54 (m, 2H), 1.17 - 1.05 (m, 4H). MS(ESI, m/z): 497[M+H] ⁺ .
White solid compound 2: yield 64%, ¹ HNMR (400 MHz, DMSO-d ₆ ) δ 8.20 - 8.16 (m, 1H), 7.70 (d, J = 8.4 Hz, 1H), 7.67 - 7.62 (m, 2H), 7.60 - 7.53 (m, 1H), 7.28 - 7.21 (m, 1H), 4.28 (s,br, 2H), 3.42 - 3.40 (m, 1H), 2.38 - 2.29 (m, 1H), 1.81 - 1.69 (m, 6H), 1.60 - 1.52 (m, 2H), 1.16 - 1.05 (m, 2H). MS(ESI, m/z): 556[M+H] ⁺ .

Example 3:
Example 3 was prepared from Intermediate VII-3 according to Scheme 1, and the synthesis scheme is as follows.

After synthesizing compound VII-3 by a synthetic method of synthesizing intermediate VII-1 from raw material XIII-3, compound 3 was obtained by scheme 1, where:
White solid compound VIII-3: yield 67%, HNMR (400 MHz, DMSO-d ₆ ) δ7.68 - 7.45 (m, 4H), 6.69 (t, J = 12.0 Hz, 2H), 4.18 (s, MS( ESI, m/z): 497[M+H] ⁺ .
White solid compound 3: yield 38%, ¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.69 - 7.42 (m, 4H), 6.74 (d, J = 13.6 Hz, 1H), 6.63 (d, J = 8.8 Hz, 1H), 4.21 (s, 2H), 3.47 - 3.43 (m, 1H), 2.35 - 2.30 (m, 1H), 1.76 - 1.70 (m, 6H), 1.58 - 1.50 (m, 2H) , 1.20 - 1.06 (m, 4H). MS(ESI, m/z): 573[M+H] ⁺ .

Example 4:
In the production of Example 4, Compound 4 was produced from Intermediate IV-1 according to Scheme 2 with reference to the operation of Example 3, and the synthesis scheme is as follows.

Lithium aluminum hydride (420 mg, 10 mmol) was placed in tetrahydrofuran (8 mL), cooled to 0°C, and a solution of compound IV-1 (2 mmol) in tetrahydrofuran (2 mL) was added drop by drop (internally). The reaction solution was stirred at room temperature for 2 h so that the temperature was below 5°C. After quenching the reaction by adding ice water (0.4 mL) at 0°C, add one drop each of 15% aqueous sodium hydroxide solution (0.4 mL) and ice water (1.2 mL), and then add anhydrous magnesium sulfate (8 g). ), and the mixture was stirred at room temperature for 0.5 h, filtered, concentrated, and subjected to column chromatography to obtain Intermediate X-1 (1 g, yield 84%). MS(ESI, m/z): 284[M+H] ⁺ .

Intermediate X-1 (1 g, 3.53 mmol) was placed in dichloromethane (20 mL), and pyridinium chlorochromate (14.14 mmol) was added at room temperature, and the reaction solution was stirred at room temperature for 1 h. Intermediate XI-1 (870 mg, 88% yield) was obtained by filtration, concentration, and column chromatography. MS(ESI, m/z): 282[M+H] ⁺ .
Lithium aluminum hydride (260 mg, 6.2 mmol) was placed in tetrahydrofuran (4 mL), cooled to 0°C, and a solution of compound XI-1 (3.1 mmol) in tetrahydrofuran (1 mL) was added dropwise (internally). The reaction solution was stirred at room temperature for 2 h so that the temperature was below 5°C. Quench the reaction by adding ice water (0.2 mL) at 0°C, then add one drop each of 15% aqueous sodium hydroxide solution (0.2 mL) and ice water (0.6 mL), and then add anhydrous magnesium sulfate (10 g). ), and the mixture was stirred at room temperature for 0.5 h, filtered, concentrated, and subjected to column chromatography to obtain Intermediate V-2 (730 mg, 83% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.62-7.60 (m, 2H), 7.56-7.52 (m, 1H), 4.92 (d, J = 5.2 Hz, 1H), 4.18 (d, J = 5.2 Hz, 1H), 2.35 - 2.28 (m, 1H), 1.14 - 1.04 (m, 4H). MS(ESI, m/z): 285[M+H] ⁺ .

Intermediate V-2 (730 mg, 2.57 mmol) was dissolved in dichloromethane (10 mL), cooled to 0 °C, phosphorus tribromide (3.08 mmol) was slowly added dropwise to the solution, and the reaction solution was incubated at room temperature for 2 h. Stirred. The solvent was removed from the reaction solution to obtain an oil, which was then charged with ethyl acetate (20 mL), the pH value of the reaction solution was adjusted to neutral with saturated aqueous sodium bicarbonate solution, washed with water, and acetic acid was added. The aqueous phase was extracted with ethyl (3 times, 100 mL each). The organic phases were mixed, washed with saturated brine, and then concentrated. Intermediate VI-2 (720 mg, yield 81%) was obtained by column chromatography. MS(ESI, m/z): 347[M+H] ⁺ .

Potassium t-butoxide (4.16 mmol) was added to a solution of compound VII-2 (512 mg, 2.08 mmol) in anhydrous tetrahydrofuran (10 mL) at 0°C, and after stirring for 15 minutes, compound VII-2 (2.08 mmol) was added dropwise. ) in anhydrous tetrahydrofuran (5 mL) was added, and the reaction mixture was stirred at room temperature for 4 h. Water (20 mL) was added to the reaction mixture, extracted with ethyl acetate (20 mL each, three times in total), the organic phase was washed with saturated brine, concentrated, and the intermediate VIII-4 (420 mg, yield 39%). MS(ESI, m/z): 513[M+H] ⁺ .
Intermediate VIII-4 (420 mg, 0.82 mmol), hydroxyamine hydrochloride (1.64 mmol), and absolute ethanol (5 mL) were placed in a round bottom flask and stirred, triethylamine (1.64 mmol) was slowly added dropwise, and the mixture was heated to 80°C. The mixture was heated and reacted overnight. Cool to room temperature, remove the solvent, dissolve in dichloromethane (20 mL), wash with water and saturated brine, concentrate the organic phase, and remove intermediate IX-4 (360 mg, yield) by silica gel column chromatography. 81%). MS(ESI, m/z): 546[M+H] ⁺ .

Intermediate IX-4 (360 mg, 0.66 mmol), N,N'-carbonyldiimidazole (0.99 mmol), and 1,4-dioxane (5 mL) were placed in a round bottom flask, and then 1,8-diazabicyclo[ 5.4.0] undecene-7 (0.99 mmol) was added, heated to 100°C, and reacted for 8 hours. The reaction station was cooled to room temperature, diluted with water (10 mL), adjusted to pH approximately 3 with 1M aqueous hydrochloric acid solution, and extracted with ethyl acetate (10 mL each, three times in total). The organic phases were combined, washed with saturated brine, and concentrated, and the resulting crude product was further subjected to silica gel column chromatography to obtain the final product, Compound 4 (76.7 mg, yield 20%). ¹ HNMR (400 MHz, DMSO-d ₆ ) δ7.65-7.62 (m, 2H), 7.58 - 7.55 (m, 1H), 7.47 (t, J = 8.6 Hz, 1H), 6.72 - 6.65 (m, 2H) ), 4.22 (s, 1H), 4.17 (s, 2H), 3.39 (s, 1H), 2.36 - 2.30 (m, 1H), 1.76-1.73 (m, 6H), 1.52 (d, J = 14.8 Hz, 2H), 1.14 - 1.08 (m, 4H). MS(ESI, m/z): 572[M+H] ⁺ .

Example 5:
Example 5 was prepared from Intermediate VII-4 according to Scheme 1, and the synthesis scheme is as follows.

After synthesizing compound VII-4 by the synthetic method of synthesizing intermediate VII-1 from raw material XIII-4, compound 5 was obtained according to scheme 1, where:
Compound VIII-5 as a white solid: yield 47%, ¹ HNMR (400 MHz, DMSO-d ₆ ) δ7.68 - 7.61 (m, 2H), 7.60 - 7.55 (m, 1H), 7.42 (d, J = 8.4 Hz, 1H), 6.71 (s, br, 1H), 6.62 (d, J = 8.8 Hz, 1H), 4.17 (s, 2H), 3.41 - 3.36 (m, 1H), 2.39 - 2.26 (m, 4H) ), 1.84 - 1.66 (m, 6H), 1.58 - 1.46 (m, 2H), 1.19 - 1.02 (m, 4H). MS(ESI, m/z): 510[M+H] ⁺ .
White solid compound 3: yield 11%, ¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.69 - 7.62 (m, 2H), 7.60 - 7.35 (m, 2H), 6.75 - 6.545 (m, 2H) ), 4.19 (s, 2H), 3.43 - 3.33 (m, 1H), 2.41 - 2.29 (m, 4H), 1.88 - 1.66 (m, 6H), 1.60 - 1.46 (m, 2H), 1.21 - 1.03 (m , 4H). MS(ESI, m/z): 569[M+H] ⁺ .

Example 6:
Example 6 was prepared from Intermediate VII-5 according to Scheme 1, and the synthesis scheme is as follows.

After synthesizing compound VII-5 by a synthetic method of synthesizing intermediate VII-1 from raw material XIII-5, compound 6 was obtained according to scheme 1, where:
White solid compound VIII-6: yield 32%, ¹ HNMR (400 MHz, DMSO-d ₆ ) δ7.66 - 7.61 (m, 2H), 7.60 - 7.52 (m, 1H), 7.32 (d, J = 8.4 Hz, 1H), 6.39 - 6.29 (m, 2H), 4.22 (s, 2H), 3.83 (s, 3H), 3.41 - 3.32 (m, 1H), 2.36 - 2.28 (m, 1H), 1.84 - 1.66 (m, 6H), 1.60 - 1.49 (m, 2H), 1.20 - 1.02 (m, 4H). MS(ESI, m/z): 526[M+H] ⁺ .
White solid compound 3: yield 9%, ¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.69 - 7.52 (m, 3H), 7.37 - 7.29 (m, 1H), 6.45 - 6.36 (m, 2H) ), 4.28 (s, 2H), 3.87 (s, 3H), 3.43 - 3.32 (m, 1H), 2.42 - 2.27 (m, 1H), 1.87 - 1.64 (m, 6H), 1.61 - 1.47 (m, 2H) ), 1.21 - 1.02 (m, 4H). MS(ESI, m/z): 585[M+H] ⁺ .

Example 7:
Example 7 was prepared from Intermediate VII-6 according to Scheme 1, and the synthesis scheme is as follows.

After synthesizing compound VII-6 by a synthetic method of synthesizing intermediate VII-1 from raw material XIII-6, compound 7 was obtained according to scheme 1, where:
Compound VIII-7 as a white solid: yield 56%, ¹ HNMR (400 MHz, DMSO-d ₆ ) δ7.75 (d, J = 9.2 Hz, 1H), 7.67 - 7.52 (m, 3H), 7.08 (s , 1H), 7.02 (d, J = 8.8 Hz, 1H), 4.32 (s, 2H), 3.43 - 3.32 (m, 1H), 2.36 - 2.28 (m, 1H), 1.78 - 1.69 (m, 3H), 1.61 - 1.50 (m, 2H), 1.16 - 1.06 (m, 4H). MS(ESI, m/z): 564[M+H] ⁺ .
White solid compound 7: yield 49%, ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.67 - 7.47 (m, 4H), 7.11 - 7.04 (m, 2H), 4.27 (s, 2H), 3.48 - 3.44 (m, 1H), 2.35 - 2.31 (m, 1H), 1.85 - 1.69 (m, 6H), 1.62 - 1.48 (m, 2H), 1.20 - 1.01 (m, 4H). MS(ESI, m/ z): 623[M+H] ⁺ .

Example 8:
Example 8 was produced from raw material II-2 according to Scheme 1, and the synthesis scheme is as follows.

here,
Colloidal compound V-3: yield 98%, ¹ HNMR (400 MHz, DMSO-d ₆ ) δ7.88 (d, J = 7.6 Hz, 1H), 7.81 - 7.71 (m, 2H), 7.59 (d, J = 7.2 Hz, 1H), 4.90 (s, 1H), 2.33-2.26 (m, 1H), 1.12-1.05 (m, 4H). MS(ESI, m/z): 286[M+H] ⁺ .
Compound VIII-8 as a white solid: yield 31%, ¹ HNMR (400 MHz, DMSO-d ₆ ) δ7.90 (d, J = 7.6 Hz, 1H), 7.81-7.72 (m, 2H), 7.58 (d , J = 7.6 Hz, 1H), 7.50 (t, J = 8.4 Hz, 1H), 6.74 (dd, J = 13.8, 2.2 Hz, 1H), 6.63 (dd, J = 8.6, 2.2 Hz, 1H), 4.20 (s, 1H), 3.61 - 3.57 (m, 1H), 3.40 - 3.32 (m, 1H), 2.33 - 2.29 (m, 1H), 1.76-1.69 (m, 6H), 1.54 (d, J = 14.4 Hz , 2H), 1.18 - 1.05 (m, 4H). MS(ESI, m/z): 514[M+H] ⁺ .
White solid compound 8: yield 29%, ¹ HNMR (400 MHz, DMSO-d ₆ ) δ7.90 (d, J = 8.0 Hz, 1H), 7.81 - 7.72 (m, 2H), 7.58 (d, J = 7.2 Hz, 1H), 7.47 (t, J = 8.6 Hz, 1H), 6.72 - 6.66 (m, 2H), 4.17 (s, 2H), 3.38 (s, 1H), 2.34 - 2.28 (m, 1H) , 1.77 - 1.73 (m, 6H), 1.52 (d, J = 14.8 Hz, 2H), 1.13 - 1.05 (m, 4H). MS(ESI, m/z): 573[M+H] ⁺ .

Example 9:
Example 9 was produced from raw material II-3 according to Scheme 1, and the synthesis scheme is as follows.

here,
Colloidal compound V-4: yield 76%, ¹ HNMR (400 MHz, DMSO-d ₆ ) δ 7.67-7.63 (m, 2H), 7.53 (d, J = 6.8 Hz, 2H), 4.96 (s, 1H) ), 2.32-2.27 (m, 1H), 1.13-1.04 (m, 4H). MS(ESI, m/z): 302[M+H] ⁺ .
Colloidal compound VIII-9: 47% yield, ¹ HNMR (400 MHz, DMSO-d ₆ ) δ 7.69 - 7.61 (m, 2H), 7.56 - 7.49 (m, 3H), 6.75 (d, J = 14.0 Hz , 1H), 6.64 (d, J = 8.8 Hz, 1H), 4.21 (s, 2H), 3.45 - 3.43 (m, 1H), 2.36 - 2.29 (m, 1H), 1.77 - 1.69 (m, 6H), 1.56 (d, J = 14.8 Hz, 2H), 1.23 - 1.06 (m, 4H). MS(ESI, m/z): 530[M+H] ⁺ .
White solid compound 8: yield 21%, ¹ HNMR (400 MHz, DMSO-d ₆ ) δ7.69 - 7.62 (m, 2H), 7.56 - 7.46 (m, 3H), 6.72 - 6.66 (m, 2H) , 4.18 (s, 2H), 3.43 (s, 1H), 2.35 - 2.31 (m, 1H), 1.80 - 1.75 (m, 6H), 1.54 (d, J = 14.4 Hz, 2H), 1.13 - 1.06 (m , 4H). MS(ESI, m/z): 589[M+H] ⁺ .

Examples of pharmacological experiments:
Methods for detecting FXR agonistic activity of compounds by methods based on reporter gene activity detection:
1.1 Construction and production of plasmids pGAL4-FXR-LBD and pG5-Luc The pGAL4-FXR-LBD and pG5-Luc plasmids used in the reporter gene detection system were constructed by conventional molecular cloning methods. The main step is to insert the cDNA sequence of FXR (NM_001206979.2) corresponding to the amino acid sequence of FXR-LBD (212-476AA) into the BamHI and NotI enzyme cleavage sites of the pGAL4 vector using PCR technology, and - Got LBD. pG5-Luc and phRL-TK plasmids were a gift from the Shanghai Institute of Drug Research, Chinese Academy of Sciences. The plasmid was transformed into DH5α E. coli using the CaCl ₂ method, further cultured and amplified, and then purified using a plasmid extraction kit (TIANGEN, #D107) to obtain the corresponding plasmid DNA.

Co-transfection of plasmids into HEK293T cells and treatment with compounds HEK293T cells were seeded in 96-well plates at a density of 1×10 ⁴ /well the day before plasmid transfection. Cells were transfected according to the instructions for the transfection reagent FuGENE® HD (Promega, #E2311). The main steps are to take one well as an example, add plasmids pGAL4-FXR-LBD, pG5-Luc and phRL-TK to 10 μL of Opti-MEM™ I medium at the ratio of 20 ng, 50 ng and 5 ng. (Gibco, #11058021) and mixed uniformly. Further, 0.25 μL of FuGENE (registered trademark) HD was added, mixed uniformly, and left at room temperature for 5 min. Additionally, 10 μL of this mixture was placed into cell wells containing 100 μL of culture medium. 6 h after co-transfection of cells, compounds were diluted in a 3-fold gradient to a maximum concentration of 1 μM and treated in cell culture medium for 24 h at a total of 10 concentrations, with a total of 2 duplicate wells. , LJN452 compound served as a positive control.

1.3 Detection with Dual-Glo Luciferase Cells were treated with compounds for 24 h and then detected according to the instructions of the Dual-Glo® Luciferase Assay System (Promega, #E2940). The main steps were aspirating and discarding 50 μL of culture solution from each well, adding an additional 50 μL of Dual-Glo® luciferase reagent, and shaking for 10 min at room temperature. 80 μL of the digestion reaction solution was placed in a white opaque optiPlate-96 well plate, and the luminescence signal value of Firefly luciferase (Firefly-Luc) was detected using an MD i3x multilabel plate reader. Furthermore, 40 μL of Dual-Glo (registered trademark) Stop & Glo (registered trademark) reagent was added, and the mixture was shaken at room temperature for 10 min. Furthermore, the luminescent signal value of Renilla luciferase (Renilla-Luc) was detected using an MD i3x multilabel plate reader. The value of the Firefly-Luc/Renilla-Luc ratio was taken as the FXR agonistic activity of the compound, and the normalization process was performed using the ratio value of the solvent DMSO group, and a dose-response curve was fitted with 4 parameters using GraphPad Prism 6.0 software. and calculated the _EC50 value.

2. Results From the experimental data, all of the compounds have some degree of FXR agonist activity, and among them, the EC ₅₀ of Examples 1, 2, 3, and 4 are all less than 5 nM, and they have very strong FXR agonist activity. I understand that. FXR agonist activity data for other Examples are shown in Table 1.
＊＊＊＊: EC ₅₀ (nM) <5; ＊＊＊: 5 < EC ₅₀ (nM) <10; **: 10 < EC ₅₀ (nM) <50; *: 50 < EC ₅₀ (nM).

The results show that the compounds of the present invention exhibit superior cellular level activity over the existing FXR agonist compound LJN452 and the deuterated control 1. In particular, the compound of Example 3 of the present application was deuterated based on the structure of the deuterium-free control product 1, and the activity was significantly improved, indicating that the relevant site is the main deuterium of such a compound. It was suggested that this was a hydrogenation site.

Pharmacokinetic Evaluation The bioavailability and pharmacokinetic behavior of deuterated Example 8 and non-deuterated Control 2 were compared in a mouse model. For each group, six male ICR mice of similar body weight were selected, in which three mice received a single oral dose of 3 mg/kg and three mice received a single dose of 1 mg/kg. It was administered intravenously in multiple doses. Blood samples were collected at 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, and 7 hours post-dose, and the concentration of plasma samples was analyzed by LC-MS/MS, and using PKSolver's free tool and non-compartmental The pharmacokinetic parameters of the compounds were analyzed by modeling (NCA) software.

Experimental plan:
Experimental compounds: Control 2 and Example 8

Experimental animals:
Twelve healthy male ICR mice were purchased from Charles River with Animal Production Permit Number: 20211231Abzz0619000376 and divided into 4 groups of 3 mice each.
Preparation of dosage form: A predetermined amount of compound was weighed and placed in 2% DMSO+15% Solutol+83% saline to prepare a clear solution.
Dosage: ICR mice were fasted overnight and each compound was administered at a dose of 3 mg/kg orally or 1 mg/kg intravenously. The administration volumes for oral and intravenous administration were 10 mL/kg and 5 mL/kg, respectively. The animals were fasted for up to 2 hours after administration.

Sample collection: Approximately 30 μL of blood was collected from the greater saphenous vein at 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, and 7 hours after administration. After placing the blood into a commercially available test tube containing _K2 -EDTA, centrifuge the blood sample for 5 min at 4600 rpm at approximately 4 °C to obtain the plasma sample, then place all plasma samples on dry ice. The samples were quickly frozen and kept at -70°C until LC-MS/MS analysis.

Sample preparation: 10 μL of plasma sample was precipitated using methanol containing 50 nmol/L dry ice naphthoflavone as an internal standard, and the mixture was thoroughly mixed and centrifuged at 14000 rpm for 5 min at 4 °C, followed by 75 min. μL of supernatant was taken and mixed with 75 μL of methanol and used for LC-MS/MS analysis.
The results of pharmacokinetic parameters are shown in Table 1.

Conclusion: The compound of Example 8 has better absorption and higher bioavailability than the compound of Control 2 and can be submitted for further studies.
All documents relating to the present invention are incorporated by reference in this application, as if each document were individually cited. Further, after reading the above contents of the present invention, those skilled in the art may make various changes and modifications to the present invention, but equivalent forms thereof shall be included within the scope of the claims of the present invention. should be understood.

Claims (10)

Compounds represented by general formula I, or enantiomers, diastereomers, tautomers, racemates, hydrates, solvates , or pharmaceutically acceptable salts thereof
(however,
Ar is a substituted or unsubstituted C ₆ -C ₁₀ aryl group, a substituted or unsubstituted 5-9 membered heteroaromatic ring (including a single ring or a fused ring, and is selected from 1-3 oxygen, sulfur and nitrogen) containing heteroatoms).
A is a substituted or unsubstituted C ₆ -C ₁₀ aryl group, a substituted or unsubstituted 5-9 membered heteroaromatic ring (including a single ring or fused rings, and 1-3 atoms selected from oxygen, sulfur and nitrogen) containing heteroatoms).
R ¹ is a substituted or unsubstituted C ₁ -C ₆ alkyl group, a substituted or unsubstituted C ₃ -C ₆ cycloalkyl group, a substituted or unsubstituted 5-9 membered heterocycle (1-3 oxygen atoms, containing a heteroatom selected from sulfur and nitrogen).
X is selected from the group consisting of hydrogen and deuterium.
Here, the above substitution means that one or more hydrogen atoms in the group are each independently halogen, C ₁ -C ₆ haloalkyl group, C ₁ -C ₆ haloalkoxy group, C ₁ -C ₆ alkyl group, C It is substituted with a substituent selected from the group consisting of ₁ -C ₆ alkoxy group, C ₃ -C ₆ cycloalkyl group, C ₃ -C ₆ cycloalkyloxy group, cyano group and nitro group. ). The above R ¹ is selected from the group consisting of a substituted or unsubstituted C ₁ -C ₆ alkyl group and a substituted or unsubstituted C ₃ -C ₆ cycloalkyl group, where the above substitution refers to a substitution in the group. One or more hydrogen atoms each independently represent a halogen, a C ₁ -C ₆ haloalkyl group, a C ₁ -C ₆ haloalkoxy group, a C ₁ -C ₆ alkyl group, a C ₁ -C ₆ alkoxy group, a C ₃ -C _6. The compound according to claim 1, wherein the compound is substituted with a substituent selected from the group consisting of a 6-cycloalkyl group, a _C3 - _C6 cycloalkyloxy group, a cyano group, and a nitro group. The above Ar is selected from the group consisting of a substituted or unsubstituted C ₆ -C ₁₀ aryl group, a substituted or unsubstituted 5-9 membered heteroaromatic ring, and the above substituent is hydrogen, fluorine, chlorine, Claim 1, characterized in that it is selected from the group consisting of bromine, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, trifluoromethyl group, and trifluoromethoxy group. compound. The above A is selected from the group consisting of a substituted or unsubstituted C ₆ -C ₁₀ aryl group, a substituted or unsubstituted 5-9 membered heteroaromatic ring, where the substitution of the above aryl group or heteroaryl group The group is selected from the group consisting of hydrogen, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, trifluoromethyl, and trifluoromethoxy. 2. The compound according to claim 1, characterized in that Claim 1, wherein R ¹ is selected from the group consisting of methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl, cyclopropyl group, cyclobutyl group, and cyclopentyl group. Compounds described in. Compound according to claim 1, characterized in that said compound is selected from the group:
A method for producing a compound according to claim 1, characterized in that it comprises the step of producing a compound of formula I by the method according to scheme 1 or scheme 2 below:
(a ) Using the compound represented by general formula II as a starting material, it is reacted with hydroxyamine hydrochloride under the action of a base to obtain an intermediate, which is then salified with N-chlorosuccinimide (NCS) to form general formula III. become the represented compound;
(b) then reacting the compound of general formula III with the corresponding 3-oxopropionic ester under the action of a base to obtain a compound of general formula IV;
(c) reducing the ester compound of the general formula IV to the corresponding alcohol, i.e. the compound of the general formula V, under the action of a deuterated reducing agent;
(d) brominating the compound represented by general formula V with a bromine reagent to produce a compound represented by general formula VI;
(e) reacting a compound represented by general formula VI and a compound represented by general formula VII under the action of a base to form a compound represented by general formula VIII;
(f) reacting a compound of general formula VIII with hydroxyamine hydrochloride under the action of a base to produce a compound of general formula IX;
(g) reacting a compound of general formula IX under the action of phosgene, triphosgene or carbonyldiimidazole to produce a compound of general formula I;
(However, X is deuterium, and the definitions of R ¹ , Ar, and A are as described in claim 1.)
(a) reducing the ester compound of the general formula IV to the corresponding alcohol, i.e. the compound of the general formula X, under the action of a reducing agent;
(b) oxidizing the compound of general formula X to the corresponding aldehyde, i.e. the compound of general formula XI, under the action of an oxidizing agent;
(c) reducing the ester compound of general formula XI to the compound of general formula V under the action of a deuterated reducing agent;
(d) brominating the compound represented by general formula V with a bromine reagent to produce a compound represented by general formula VI;
(e) reacting a compound represented by general formula VI and a compound represented by general formula VII under the action of a base to form a compound represented by general formula VIII;
(f) reacting a compound of general formula VIII with hydroxyamine hydrochloride under the action of a base to produce a compound of general formula IX;
(g) reacting a compound represented by general formula IX under the action of phosgene, triphosgene or carbonyldiimidazole to produce a compound represented by general formula I (in each formula, X is hydrogen, R ¹ , The definitions of Ar and A are as described in claim 1). A compound represented by the general formula I according to claim 1, or an enantiomer, diastereomer, tautomer, racemate, hydrate, solvate , or pharmaceutically acceptable salt thereof; , and a pharmaceutically acceptable carrier. The compound represented by the general formula I according to claim 1, or its enantiomer, diastereomer, tautomer, racemate, hydrate, solvate, or a pharmaceutically acceptable salt thereof The use is characterized in that it is used in the manufacture of a pharmaceutical composition for treating a disease or disease related to the activity or expression level of FXR. Use according to claim 9, characterized in that said FXR-related disease is selected from the group consisting of diseases associated with processes of bile acid metabolism, carbohydrate metabolism, lipid metabolism, inflammation and/or liver fibrosis. .