CN112876493B

CN112876493B - Salts of nitrogen-containing tricyclic compounds and uses thereof

Info

Publication number: CN112876493B
Application number: CN202011353544.8A
Authority: CN
Inventors: 陈亮; 单岳峰; 王晓军; 杨新业; 梁小小; 宗乔; 崔艳慧
Original assignee: Sunshine Lake Pharma Co Ltd
Current assignee: Guangdong HEC Pharmaceutical
Priority date: 2019-11-29
Filing date: 2020-11-27
Publication date: 2023-05-09
Anticipated expiration: 2040-11-27
Also published as: CN112876493A

Abstract

The present invention relates to salts of nitrogen-containing tricyclic compounds and uses thereof. The invention also relates to a pharmaceutical composition comprising said salt or thereof, and to the use of said salt or said pharmaceutical composition for the preparation of a medicament for preventing, treating or alleviating FXR mediated diseases in a patient.

Description

Salts of nitrogen-containing tricyclic compounds and uses thereof

Technical Field

The invention belongs to the technical field of medicaments, relates to salts of nitrogenous tricyclic compounds and application thereof, in particular to salts of 2- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -10H-spiro [ benzo [6,7] oxazepin [3,2-b ] pyridine-11, 1' -cyclopropane ] -7-carboxylic acid (a compound shown as a formula (I)) and application thereof, and further relates to a pharmaceutical composition containing the salts. The salts or the pharmaceutical compositions are useful for preventing, treating or alleviating FXR mediated diseases in a patient.

Background

The Farnesol X Receptor (FXR) is a member of the nuclear hormone receptor superfamily and is expressed primarily in the liver, kidneys and intestines (Seol et al, mol. Endocrinol (1995), 9:72-85; forman et al, cell (1995), 81:687-693). It acts as a heterodimer with retinoic acid X receptor (RXR) and binds to a response element in the target gene promoter to regulate gene transcription. FXR-RXR heterodimers bind with highest affinity to the inverted repeat-1 (IR-1) response element, where the hexamer binding to the consensus receptor (consensus receptor) is separated by a nucleotide. FXR can be activated by bile acids (end products of cholesterol metabolism) (Makishima et al Science (1999), 284:1362-1365; parks et al Science (1999), 284:1365-1368; wang et al, moI.cell. (1999), 3:543-553), whereas bile acids are used to inhibit catabolism of cholesterol (Urizar et al, (2000) J.biol. Chem. 275:39313-393170).

FXR is a key regulator of cholesterol homeostasis, triglyceride synthesis, and adipogenesis (Crawley, expert Opinion Ther.patents (2010), 20:1047-1057). In addition to being useful as targets for the treatment of dyslipidemia, obesity, vitamin D-related diseases, intestinal disorders, drug-induced side effects, and hepatitis (Crawley, expert open Ther. Patents (2010), 20:1047-1057), FXR is also useful as a therapeutic target for hepatobiliary diseases, chronic hepatitis, non-alcoholic fatty liver (NAFLD), non-alcoholic steatohepatitis (NASH), cholestasis, liver fibrosis, cirrhosis, hepatitis B, metabolic disorders, lipometabolic disorders, carbohydrate metabolic disorders, cardiovascular metabolic disorders, atherosclerosis, type II diabetes, and diabetic complications (Frank G.Schaap et al, journal of Medicinal Chemistry (2005), 48:5383-5402).

Patent applications WO 2018024224 and CN 107686486 disclose nitrogen-containing tricyclic compounds useful as modulators of FXR activity, and methods for their preparation and use, wherein compound 7, namely compound 2- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -10H-spiro [ benzo [6,7] oxa [3,2-b ] pyridine-11, 1' -cyclopropane ] -7-carboxylic acid (compound of formula (I)), is specifically disclosed. However, this patent application does not disclose any salts of the compounds or specific microstructures thereof.

It is well known in the art that drug free base or free acid generally has poor solubility and thus low bioavailability problems, which can be generally solved by converting it into a salt, however, different salts and solid forms of drug free base or free acid may have different properties. Modification of properties resulting from different salt or solid forms may also improve the final dosage form, for example, if such modification may increase bioavailability. Therefore, in drug development, research to find a suitable salt for drug utilization is very important.

Disclosure of Invention

The invention provides a base addition salt of a compound shown in a formula (I) and a pharmaceutical composition thereof, wherein the base addition salt or the pharmaceutical composition has better pharmacological properties (such as good pharmacokinetic properties), and the stability and other properties of the base addition salt or the pharmaceutical composition are obviously improved, so that the base addition salt or the pharmaceutical composition has better patentability.

In particular, the invention relates to a base addition salt of a compound shown in formula (I) and a pharmaceutical composition thereof, and application of the base addition salt or the pharmaceutical composition in preparing medicines for preventing, treating or relieving FXR-mediated diseases of patients. The base addition salts of the present invention may also be in the form of solvates, for example hydrates.

In one aspect, the invention provides pharmaceutically acceptable base addition salts of compounds of formula (I),

in some embodiments, the base addition salts shown in the present invention are inorganic and organic base salts.

In other embodiments, the inorganic base salts of the present invention are lithium salts, sodium salts, potassium salts, calcium salts, magnesium salts, or any combination thereof.

In other embodiments, the organic base salt of the present invention is an ammonium salt, choline salt, lysine salt, arginine salt, ethanolamine salt, dimethylethanolamine salt, triethanolamine salt, tromethamine salt, N-methylglucamine salt, morpholine salt, piperazine salt, t-butylamine salt, dicyclohexylamine salt, or any combination thereof.

In some embodiments, the base addition salts of the compounds of formula (I) of the present invention are amorphous in form, wherein the amorphous form of the potassium salt has an X-ray powder diffraction pattern substantially as shown in figure 1.

In some embodiments, the ethanolamine salt of the compound of formula (I) of the base addition salts of the present invention is amorphous, characterized by an X-ray powder diffraction pattern substantially as shown in figure 2.

In some embodiments, the dimethylethanolamine salt of the compound of formula (I) of the base addition salts described herein is amorphous, characterized in that the dimethylethanolamine salt is amorphous with an X-ray powder diffraction pattern substantially as depicted in figure 3.

In some embodiments, the base addition salts of the compounds of formula (I) of the present invention are amorphous in triethanolamine salts, characterized in that the triethanolamine salts are amorphous having an X-ray powder diffraction pattern substantially as shown in figure 4.

In some embodiments, the base addition salts of the compounds of formula (I) of the present invention are amorphous in form, wherein the amorphous form of the tromethamine salt has an X-ray powder diffraction pattern substantially as shown in figure 5.

In one aspect, the invention also provides a pharmaceutical composition comprising any of the base addition salts described herein, or a combination thereof, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, or combination thereof.

In a further aspect, the invention also relates to the use of a base addition salt of a compound of formula (I) or of the pharmaceutical composition for the manufacture of a medicament for the prevention, treatment or alleviation of FXR mediated diseases in a patient; further, the use comprises administering to a human or animal a therapeutically effective amount of a base addition salt of the invention or of the pharmaceutical composition.

In some embodiments, the FXR mediated disease described herein is cardiovascular and cerebrovascular disease, a disease associated with dyslipidemia, metabolic syndrome, hyperproliferative disease, fibrosis, inflammatory disease, or a disease associated with liver and gall.

In other embodiments, the cardiovascular and cerebrovascular diseases described herein are atherosclerosis, acute myocardial infarction, venous occlusive diseases, portal hypertension, pulmonary hypertension, heart failure, peripheral arterial occlusive diseases, sexual dysfunction, stroke, or thrombosis.

In other embodiments, the metabolic syndrome of the invention is insulin resistance, hyperglycemia, hyperinsulinemia, elevated levels of fatty acids or triglycerides in the blood, hyperlipidemia, obesity, hypertriglyceridemia, hypercholesterolemia, syndrome X, diabetic complications, atherosclerosis, hypertension, acute anemia, neutropenia, dyslipidemia, type II diabetes, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, dyslipidemia, or a combination of diabetes and abnormally high body mass index.

In other embodiments, the hyperproliferative diseases described herein are hepatocellular carcinoma, colon adenoma, polyposis, colon adenocarcinoma, breast cancer, membranous adenocarcinoma, butcher's esophageal cancer, and other forms of gastrointestinal or liver neoplastic diseases.

In other embodiments, the fibrosis, inflammatory disease, or disease associated with liver and gall is non-alcoholic fatty liver, non-alcoholic steatohepatitis, cholestasis, liver fibrosis, primary biliary cirrhosis, primary sclerosing cholangitis, progressive familial cholestasis, cystic fibrosis, drug-induced biliary damage, gallstones, cirrhosis, hepatitis b, sebaceous gland disorders, alcohol-induced cirrhosis, biliary obstruction, cholelithiasis, colitis, neonatal Huang Zheng, yellow-core disease, or intestinal bacterial overgrowth.

In one aspect, the invention relates to a method for preventing, treating or alleviating FXR mediated diseases in a patient comprising administering to the patient a pharmaceutically acceptable effective dose of a base addition salt according to the invention or of a pharmaceutical composition according to the invention.

In another aspect, the invention relates to the use of a base addition salt of a compound of formula (I) or of the pharmaceutical composition for preventing, treating or alleviating FXR mediated diseases in a patient.

In another aspect, the present invention also relates to a process for the preparation of a base addition salt of a compound of formula (I).

The solvent used in the process for producing a base addition salt according to the present invention is not particularly limited, and any solvent which dissolves the starting materials to a certain extent and does not affect the properties thereof is included in the present invention. In addition, many similar modifications, equivalent substitutions, or equivalent solvents, combinations of solvents, and different proportions of solvent combinations described herein are considered to be encompassed by the present invention. The present invention gives the preferred solvents to be used in each reaction step.

The preparation experiments of the base addition salts according to the invention will be described in detail in the examples section. Meanwhile, the invention provides activity test experiments of the base addition salt, such as a pharmacokinetics experiment, a stability experiment, a hygroscopicity experiment and the like. The experimental result shows that the base addition salt of the compound shown in the formula (I) has better bioactivity and high stability, and is suitable for pharmaceutical use. In particular, the base addition salts of the present invention have more excellent pharmaceutical properties, e.g., higher exposure.

Wherein, regarding the definition of the characteristic description of the hygroscopicity and the weight gain of the hygroscopicity (the guiding principle of the drug hygroscopicity test in appendix 9103 of 2015 of Chinese pharmacopoeia, the experimental conditions are 25 ℃ +/-1 ℃ and 80% +/-2% relative humidity) are shown in the following table:

characterization of hygroscopicity and definition of the weight gain of hygroscopicity

The alkali addition salt is not easy to be affected by high humidity to deliquesce, and is convenient for long-term storage and placement of medicines.

Definitions and general terms

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All patents and publications referred to herein are incorporated by reference in their entirety. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods, devices, and materials are described herein.

"crystalline form" or "crystalline form" refers to a solid having a highly regular chemical structure, including, but not limited to, single or multicomponent crystals, and/or polymorphs, solvates, hydrates, clathrates, co-crystals, salts, solvates of salts, hydrates of salts of the compounds. The crystalline form of a substance may be obtained by a number of methods known in the art. Such methods include, but are not limited to, melt crystallization, melt cooling, solvent crystallization, crystallization in a defined space, e.g., in a nanopore or capillary, crystallization on a surface or template, e.g., on a polymer, crystallization in the presence of additives such as co-crystallizing anti-molecules, desolvation, dehydration, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, sublimation, reactive crystallization, anti-solvent addition, milling, solvent drop milling, and the like.

"amorphous" or "amorphous form" refers to a substance that forms when particles (molecules, atoms, ions) of the substance are non-periodically arranged in three dimensions, characterized by a diffuse, non-spiking X-ray powder diffraction pattern. Amorphous is a special physical form of solid material whose locally ordered structural features suggest a myriad of interactions with crystalline material. Amorphous forms of a substance can be obtained by a number of methods known in the art. Such methods include, but are not limited to, quenching, antisolvent flocculation, ball milling, spray drying, freeze drying, wet granulation, and solid dispersion techniques, among others.

"solvent" refers to a substance (typically a liquid) that is capable of completely or partially dissolving another substance (typically a solid). Solvents useful in the practice of the present invention include, but are not limited to, water, acetic acid, acetone, acetonitrile, benzene, chloroform, carbon tetrachloride, methylene chloride, dimethyl sulfoxide, 1, 4-dioxane, ethanol, ethyl acetate, butanol, tertiary butanol, N-dimethylacetamide, N-dimethylformamide, formamide, formic acid, heptane, hexane, isopropanol, methanol, methyl ethyl ketone, mesitylene, nitromethane, polyethylene glycol, propanol, pyridine, tetrahydrofuran, toluene, xylene, mixtures thereof, and the like.

"solvate" means a compound having a solvent on, in, or on and in the crystal lattice that can be water, acetic acid, acetone, acetonitrile, benzene, chloroform, carbon tetrachloride, methylene chloride, dimethyl sulfoxide, 1, 4-dioxane, ethanol, ethyl acetate, butanol, t-butanol, N-dimethylacetamide, N-dimethylformamide, formamide, formic acid, heptane, hexane, isopropanol, methanol, methyl ethyl ketone, methyl pyrrolidone, mesitylene, nitromethane, polyethylene glycol, propanol, pyridine, tetrahydrofuran, toluene, xylene, mixtures thereof, and the like. A specific example of a solvate is a hydrate, wherein the solvent on the surface, in the crystal lattice or both is water. The hydrate may or may not have other solvents than water on the surface of the substance, in the crystal lattice, or both.

Crystalline forms or amorphous forms may be identified by a variety of techniques, such as X-ray powder diffraction (XRPD), infrared absorption spectroscopy (IR), melting point, differential Scanning Calorimetry (DSC), thermogravimetric analysis (TGA), nuclear magnetic resonance, raman spectroscopy, X-ray single crystal diffraction, dissolution calorimetry, scanning Electron Microscopy (SEM), quantitative analysis, solubility and dissolution rate, and the like.

The present invention relates to various base addition salts of said 2- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -10H-spiro [ benzo [6,7] oxazepin [3,2-b ] pyridine-11, 1' -cyclopropane ] -7-carboxylic acid (compound of formula (I)) which are present in substantially pure form.

In the context of the present invention, when used or whether or not the word "about" or "about" is used, means within 10%, suitably within 5%, particularly within 1% of a given value or range. Alternatively, the term "about" or "approximately" means within an acceptable standard error of the average value to one of ordinary skill in the art. Whenever a number is disclosed having a value of N, any number within the values of N+/-1%, N+/-2%, N+/-3%, N+/-5%, N+/-7%, N+/-8% or N+/-10% will be explicitly disclosed, where "+/-" means plus or minus.

"room temperature" in the present invention means a temperature from about 10℃to about 40 ℃. In some embodiments, "room temperature" refers to a temperature from about 20 ℃ to about 30 ℃; in other embodiments, "room temperature" refers to 20 ℃,22.5 ℃,25 ℃,27.5 ℃, and so forth.

Pharmaceutical compositions, formulations, administration and uses of the base addition salts of the invention

The pharmaceutical composition of the invention is characterized by comprising any base addition salt of the compound of formula (I) or any combination thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of the base addition salt of the compound in the pharmaceutical composition of the invention is effective to detectably treat or ameliorate FXR mediated diseases in a patient.

As described herein, the pharmaceutically acceptable compositions of the present invention further comprise a pharmaceutically acceptable carrier, adjuvant, or vehicle, as used herein, including any solvents, diluents, or other liquid excipients, dispersing or suspending agents, surfactants, isotonic agents, thickening agents, emulsifying agents, preservatives, solid binders or lubricants, and the like, suitable for the particular target dosage form. As described in the following documents: in Remington, the Science and Practice of Pharmacy,21st edition,2005,ed.D.B.Troy,Lippincott Williams&Wilkins,Philadelphia,and Encyclopedia of Pharmaceutical Technology,eds.J.Swarbrick and J.C.Boylan,1988-1999,Marcel Dekker,New York, in combination with the teachings of the literature herein, shows that different carriers can be used In the formulation of pharmaceutically acceptable compositions and their well-known methods of preparation. In addition to the extent to which any conventional carrier vehicle is incompatible with the compounds of the present invention or base addition salts thereof, such as any adverse biological effects produced or interactions with any other component of the pharmaceutically acceptable composition in a deleterious manner, their use is also contemplated by the present invention.

The base addition salts of the present invention may be incorporated as the active ingredient in a homogeneous mixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier may be in a wide variety of forms depending on the form of preparation required for administration, for example, orally or parenterally (including intravenously). When preparing compositions for oral dosage forms, any conventional pharmaceutical medium may be used, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like in the preparation of oral liquid medicaments such as suspensions, elixirs and solutions; or in the preparation of oral solid preparations such as powders, hard capsules, soft capsules and tablets, such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents and the like, wherein solid oral preparations are more preferable than liquid preparations.

Because tablets and capsules are easy to take, they represent the most advantageous oral dosage unit form, in which case solid pharmaceutical carriers are obviously employed. If desired, the tablets may be coated with standard aqueous or non-aqueous techniques. Such compositions and formulations should contain at least 0.1% active ingredient. Of course, the percentage of active ingredient in these compositions may vary, and this percentage may conveniently vary from about 2% to about 60% per weight. The active ingredient may be administered intranasally, for example, in the form of droplets or a spray.

The tablets, pills, capsules, and the like may also comprise: binders (such as gum tragacanth, acacia, corn starch or gelatin); excipients (such as dicalcium phosphate); disintegrants (e.g., corn starch, potato starch, alginic acid); lubricants (such as magnesium stearate); and sweeteners (such as sucrose, lactose or saccharin). When the dosage unit form is a capsule, it may contain a liquid carrier (such as a fatty oil) in addition to materials of the type described above.

A wide variety of other materials may be present as coatings or to alter the shape of the dosage unit. For example, the tablets may be coated with shellac, sugar or both. In addition to the active ingredient, a syrup or elixir may contain sucrose as a sweetening agent, methyl or propylparaben as a preservative, a dye and a flavoring (e.g., cherry or orange flavored).

Ophthalmic preparations, ophthalmic ointments, powders, solutions, and the like are also included within the scope of the present invention.

The base addition salts of the present invention may also be administered parenterally. Solutions or suspensions of these active materials can be prepared by appropriate mixing in water with a surfactant such as hydroxypropylcellulose. Dispersants may also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof, and in oils. Under conventional conditions of storage and use, these formulations contain a preservative to prevent the growth of microorganisms.

Pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the pharmaceutical form must be sterile and must be fluid in a form that is easy to inject. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier may be a solvent or dispersion medium containing, for example: water, ethanol, polyols (such as glycerol, propylene glycol and liquid polyethylene glycols), suitable mixtures thereof and vegetable oils.

Any suitable method of administration may be used to provide an effective dose of the base addition salts of the invention to mammals, particularly humans. For example, oral, rectal, topical, parenteral, ocular, pulmonary, nasal, etc. methods of administration may be used. Dosage forms include tablets, troches, dispersions, suspensions, solutions, capsules, emulsions, ointments, aerosols, and the like. Preferably, the base addition salts of the present invention are administered orally.

The therapeutically effective dose of the base addition salts, pharmaceutical compositions or combinations thereof of the present invention will depend on the species, weight, age and condition of the individual, the disorder or disease to be treated or the severity thereof. A physician, clinician or veterinarian of ordinary skill can readily determine the effective amount of each of the active ingredients required to prevent, treat or inhibit the progression of the disorder or disease.

When the compound of the present invention or a base addition salt thereof is used for the treatment or prophylaxis of the FXR mediated disorder described in the present invention, a substantially satisfactory effect is obtained when the compound of the present invention or a base addition salt thereof is administered in a daily dose of about 0.1 mg to about 100 mg per kg of animal body weight, preferably in a single daily dose, or in divided doses of 2 to 6 times per day, or in a continuous release form. For most large mammals, the total daily dose is from about 1.0mg to about 1000 mg, preferably from about 1mg to about 50 mg. For an adult of 70 kg, the total daily dose is generally from 7 mg to about 350 mg. This dosage regimen can be adjusted to provide the best therapeutic effect.

The base addition salt or the pharmaceutical composition thereof according to the present invention is useful for preventing, treating or alleviating FXR-mediated diseases in patients, and in particular, can be used for treating nonalcoholic fatty liver (NAFLD), nonalcoholic steatohepatitis (NASH), obesity, hypertriglyceridemia, atherosclerosis, chronic intrahepatic cholestasis, primary Biliary Cirrhosis (PBC), primary Sclerosing Cholangitis (PSC), progressive familial cholestasis (PFIC), drug-induced bile duct injury, cholelithiasis, cirrhosis, hepatitis b, sebaceous gland disease, alcohol-induced cirrhosis, cystic fibrosis, biliary tract blockage, cholelithiasis, hepatic fibrosis, dyslipidemia, atherosclerosis, diabetes II, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, peripheral tissue arterial occlusive disease (PAOD), colitis, neonatal Huang Zheng, core yellow disease, venous occlusive disease, portal hypertension, metabolic syndrome, acute infarction, myocardial infarction, hypercholesterolemia, thrombosis, gastrointestinal thrombosis, hypercholesterolemia, bacterial dysfunction, and other diseases of the liver, and the like.

Drawings

FIG. 1 is an X-ray powder diffraction (XRPD) pattern of the potassium salt of the compound of formula (I) prepared according to the method of example 1 of the invention.

FIG. 2 is an X-ray powder diffraction (XRPD) pattern of an ethanolamine salt of a compound of formula (I) prepared according to the process of example 2 of the present invention.

FIG. 3 is an X-ray powder diffraction (XRPD) pattern of the dimethylethanolamine salt of the compound of formula (I) produced by the process of example 3 according to the present invention.

FIG. 4 is an X-ray powder diffraction (XRPD) pattern of the triethanolamine salt of the compound of formula (I) prepared according to the method of example 4 of the present invention.

FIG. 5 is an amorphous X-ray powder diffraction (XRPD) pattern of the tromethamine salt of the compound of formula (I) prepared according to the method of example 5 of the present invention.

Detailed Description

The invention is further illustrated by way of examples which are not intended to limit the scope of the invention.

The X-ray powder diffraction analysis method used in the invention comprises the following steps: an Empyrean diffractometer was used to obtain X-ray powder diffraction patterns using Cu-K alpha radiation (45 KV,40 mA). The powdered sample was prepared as a thin layer on a monocrystalline silicon sample holder, placed on a rotating sample stage and analyzed in steps of 0.0167 ° in the range of 3 ° -60 °. Data was collected using Data Collector software, high Score Plus software processed the Data, and Data Viewer software read the Data.

Detailed description of the preferred embodiments

Specific synthetic methods for the compound 2- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -10H-spiro [ benzo [6,7] oxazepin [3,2-b ] pyridine-11, 1' -cyclopropane ] -7-carboxylic acid of formula (I) are described in patent application and in example 9 of CN 107686486.

Examples

EXAMPLE 1 amorphous potassium salt of Compound of formula (I)

1. Preparation of amorphous potassium salt

Compound 2- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -10H-spiro [ benzo [6,7] oxazepin [3,2-b ] pyridine-11, 1' -cyclopropane ] -7-carboxylic acid (211.0 mg) was dissolved in acetone (4.0 mL) at room temperature, then a solution of potassium isooctanoate (72.1 mg) in acetone (1.0 mL) was slowly added to the above solution, stirred at room temperature for 6 hours, then cooled to 5 ℃ and incubated for 30 minutes; the solvent was distilled off under reduced pressure, and the residue was dried at room temperature under vacuum for 4 hours to give a white solid (215.4 mg, 95.5%).

2. Characterization of potassium salt amorphous form

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis: using Cu-ka radiation, the potassium salt amorphous X-ray powder diffraction (XRPD) pattern is substantially as shown in figure 1, with a margin of error of ± 0.2 °.

EXAMPLE 2 ethanolamine salt of Compound of formula (I) amorphous form

1. Preparation of ethanolamine salt amorphous form

The compound 2- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -10H-spiro [ benzo [6,7] oxazepin [3,2-b ] pyridine-11, 1' -cyclopropane ] -7-carboxylic acid (676.0 mg) and ethanolamine (75.3 mg) were added to acetone (14.0 mL) at room temperature and stirred at room temperature for 6 hours, the system was clear; the solvent was evaporated under reduced pressure and the residue was dried overnight at room temperature under vacuum to give a white solid (685.0 mg, 92.9%).

2. Characterization of ethanolamine salt amorphous form

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis: using Cu-ka radiation, the ethanolamine salt amorphous X-ray powder diffraction (XRPD) pattern is substantially as shown in figure 2, with a margin of error of ± 0.2 °.

EXAMPLE 3 amorphous dimethylethanolamine salt of Compound of formula (I)

1. Amorphous preparation of dimethylethanolamine salt

The compound 2- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -10H-spiro [ benzo [6,7] oxazepin [3,2-b ] pyridine-11, 1' -cyclopropane ] -7-carboxylic acid (658.0 mg) and dimethylethanolamine (105.8 mg) was added to acetone (15.0 mL) at room temperature, stirred at room temperature for 6 hours, then the solvent was distilled off under reduced pressure and the residue was dried under vacuum overnight at 60 ℃ to give a white solid (700.0 mg, 93.0%).

2. Identification of dimethylethanolamine salt amorphous form

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis: using Cu-ka radiation, the dimethylethanolamine salt amorphous X-ray powder diffraction (XRPD) pattern is substantially as shown in figure 3, with a margin of error of ± 0.2 °.

EXAMPLE 4 amorphous triethanolamine salt of Compound of formula (I)

1. Amorphous preparation of triethanolamine salts

The compound 2- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -10H-spiro [ benzo [6,7] oxazepin [3,2-b ] pyridine-11, 1' -cyclopropane ] -7-carboxylic acid (598.0 mg) and triethanolamine (163.0 mg) was added to acetone (10.0 mL) at room temperature and stirred overnight at room temperature, the solid was not completely dissolved, ethanol (2.0 mL) was added and the system became clear; the solvent was evaporated under reduced pressure and the residue was dried under vacuum overnight to give a white solid (550.0 mg, 73.2%).

2. Characterization of triethanolamine salt amorphous form

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis: using Cu-ka radiation, the triethanolamine salt amorphous X-ray powder diffraction (XRPD) pattern is substantially as shown in figure 4, with a margin of error of ± 0.2 °.

EXAMPLE 5 amorphous tromethamine salt of Compound of formula (I)

1. Preparation of amorphous tromethamine salts

The compound 2- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -10H-spiro [ benzo [6,7] oxazepin [3,2-b ] pyridine-11, 1' -cyclopropane ] -7-carboxylic acid (608.0 mg) and tromethamine (134.2 mg) was added to ethanol (18.0 mL) at room temperature, stirred at room temperature, the solid was insoluble, heated to 85.0 ℃, the solid slowly dissolved, then cooled naturally to room temperature, and stirred overnight; the solvent was evaporated under reduced pressure and the residue was dried overnight at room temperature under vacuum to give a white solid (680.0 mg, 93.0%).

2. Identification of the amorphous form of tromethamine salt

(1) Identified by Empyrean X-ray powder diffraction (XRPD) analysis: using Cu-ka radiation, the tromethamine salt amorphous X-ray powder diffraction (XRPD) pattern is substantially as shown in figure 5, with a margin of error of ± 0.2 °.

EXAMPLE 6 pharmacokinetic experiments on base addition salts according to the invention

The base addition salt of the compound of formula (I) according to the invention (compound 2- ((5-cyclopropyl-3- (2, 6-dichlorophenyl) isoxazol-4-yl) methoxy) -10H-spiro [ benzo [6,7] oxazepin [3,2-b ] pyridine-11, 1' -cyclopropane ] -7-carboxylic acid) is encapsulated for oral administration.

8-12kg male Beagle dogs were orally administered 5mg/kg samples for each group of 3 animals. Blood was collected at time points 0.25,0.5,1.0,2.0,4.0,6.0,8.0 and 24 hours after administration. A standard curve of a suitable range is established according to the sample concentration, the concentration of the test sample in the plasma sample is determined in MRM mode using the AB SCIEX API4000 type LC-MS/MS, and quantitative analysis is performed. According to the drug concentration-time curve, the pharmacokinetic parameters were calculated using the WinNonLin 6.3 software non-compartmental model. The experimental results are shown in table 1.

TABLE 1 pharmacokinetic experimental data for the base addition salts of the invention

Conclusion of experiment:

as can be seen from Table 1, the base addition salts of the present invention have a relatively large exposure to Beagle and good pharmacokinetic properties.

EXAMPLE 7 stability test of the base addition salt according to the invention

(1)High temperature experiments: putting a batch of samples into a flat weighing bottle, spreading into a thin layer with the thickness less than or equal to 5mm, standing at 60 ℃ for 30 days, sampling at 5 th, 10 th and 30 th days, observing color change of the samples, and detecting the purity of the samples by HPLC.

(2)High humidity experiment: placing a batch of test sample into flat weighing bottle, spreading into thin layer with thickness less than or equal to 5mm, and placing under 25 deg.C and RH 90% ± 5%After 30 days, samples were taken on days 5, 10 and 30, the color change of the samples was observed, and the purity of the samples was checked by HPLC.

(3)Illumination experiment: placing a batch of test sample into flat weighing bottle, spreading into thin layer with thickness less than or equal to 5mm, placing into illumination box (with ultraviolet), and placing into illumination intensity 4500+ -500 lx, ultraviolet light not less than 0.7w/m ² The sample was left to stand for 30 days under the condition, sampled on days 5, 13 and 30, the color change of the sample was observed, and the purity of the sample was checked by HPLC.

Conclusion of experiment:

under the conditions of high temperature, high humidity and illumination, the appearance and purity of the base addition salt are not obviously changed, and the stability effect is good, so that the base addition salt is suitable for pharmaceutical use.

EXAMPLE 8 hygroscopicity test of the base addition salts of the invention

And taking a proper amount of a test sample, and testing the wettability of the test sample by adopting a dynamic moisture adsorption instrument. Experimental results prove that the alkali addition salt is not easy to be affected by high humidity and deliquesced.

The above description is merely a basic description of the inventive concept, and any equivalent transformation according to the technical solution of the present invention shall fall within the protection scope of the present invention.

In the description of the present specification, a description referring to terms "one embodiment," "some embodiments," "examples," "specific examples," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, the different embodiments or examples described in this specification and the features of the different embodiments or examples may be combined and combined by those skilled in the art without contradiction.

While embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and not to be construed as limiting the invention, and that variations, modifications, alternatives and variations may be made to the above embodiments by one of ordinary skill in the art within the scope of the invention.

Claims

1. A pharmaceutically acceptable base addition salt of a compound of formula (I),

wherein the base addition salt is tromethamine salt;

wherein the tromethamine salt is amorphous of a tromethamine salt of a compound of formula (I), characterized in that the tromethamine salt has an X-ray powder diffraction pattern substantially as shown in figure 5.

2. A pharmaceutical composition comprising the base addition salt of claim 1, and a pharmaceutically acceptable carrier, excipient, diluent, or combination thereof.

3. Use of a base addition salt according to claim 1 or a pharmaceutical composition according to claim 2 for the manufacture of a medicament for the prevention, treatment or alleviation of FXR mediated diseases in a patient.

4. The use according to claim 3, wherein the FXR mediated disease is cardiovascular and cerebrovascular disease, dyslipidemia related disease, metabolic syndrome, hyperproliferative disease, fibrosis, inflammatory disease or liver and gall related disease.

5. The use according to claim 4, wherein the cardiovascular and cerebrovascular disease is atherosclerosis, acute myocardial infarction, venous occlusive disease, portal hypertension, pulmonary hypertension, heart failure, peripheral arterial occlusive disease, sexual dysfunction, stroke or thrombosis;

the metabolic syndrome is insulin resistance, hyperglycemia, hyperinsulinemia, obesity, hypertriglyceridemia, hypercholesterolemia, syndrome X, hypertension, acute anemia, neutropenia, type II diabetes, or a combination of diabetes and abnormally high body mass index;

the hyperproliferative disease is hepatocellular carcinoma, colon adenoma, polyposis, colon adenocarcinoma, breast cancer, membranous adenocarcinoma, butcher's esophagus cancer or other forms of gastrointestinal neoplastic disease;

the fibrosis, inflammatory disease or liver and gall related disease is non-alcoholic fatty liver, non-alcoholic steatohepatitis, cholestasis, liver fibrosis, primary sclerosing cholangitis, progressive familial cholestasis, cystic fibrosis, drug-induced bile duct injury, gall stones, cirrhosis, hepatitis b, sebaceous gland disorders, biliary obstruction, colitis, neonatal Huang Zheng, yellow-core disease or intestinal bacterial overgrowth.

6. The use according to claim 4, wherein the metabolic syndrome is a diabetic complication.

7. The use of claim 4, wherein the metabolic syndrome is diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, or hyperlipidemia;

the hyperproliferative disease is liver tumor disease;

the diseases related to liver and gall are cholelithiasis, primary biliary cirrhosis or cirrhosis caused by alcohol.