TWI522358B - Tetrahydro-imidazo(1,5-a)pyrazine derivatives salts, preparation process and pharmaceutical use thereof - Google Patents

Description

Salt of tetrahydroimidazo[1,5-a]pyridinium, preparation method thereof and application thereof in medicine

The present invention relates to ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydro Imidazo[1,5-a]pyridyl The form of a pharmaceutically acceptable salt of 1-carboxylic acid, a process for its preparation, a pharmaceutical composition comprising the pharmaceutically acceptable salt, and the use thereof as a therapeutic agent, in particular as a dipeptidyl peptidase IV (DPP IV) inhibitor.

Diabetes is a very old metabolic disease, characterized by chronic hyperglycemia, which is caused by an increase in the concentration of glucose in the blood caused by absolute or relative deficiency of insulin in the human body, and then a large amount of sugar is excreted from the urine, thereby causing sugar, fat and protein. Metabolic disorders, physiological manifestations of polydipsia, polyuria, polyphagia, weight loss, dizziness, fatigue and other symptoms.

Permanent or uncontrolled hyperglycemia leads to an increase in morbidity and mortality. Often, abnormalities in glucose homeostasis are directly or indirectly related to changes in lipids, lipoproteins, lipoprotein metabolism, or other metabolic and hemodynamic diseases. Patients with type 2 diabetes have a significant increase in the incidence of large porous liposomes and microvascular syndromes such as coronary heart disease, stroke, peripheral vascular disease, hypertension, kidney disease, neuropathy and retinopathy. Therefore, the treatment and control of diseases such as constant blood sugar, lipid metabolism, and hypertension are extremely important for clinical treatment of diabetes.

Generally, there are two types of diabetes. People with type 1 diabetes, insulin-dependent diabetes mellitus (IDDM), produce little or no insulin on their own. Insulin is a hormone used in the body to regulate glucose utilization. People with type 2 diabetes, insulin-dependent diabetes mellitus (NIDDM), have similar or higher plasma insulin concentrations in patients with non-diabetic patients. However, such patients are resistant to insulin, which is the main insulin. Glucose and lipid metabolism in sensitive tissue cells such as muscle, liver, and 25 adipose tissue are stimulating. Even if the plasma insulin concentration is increased, it is impossible to overcome the patient's significant resistance to insulin.

Insulin resistance is mainly caused by a decrease in the number of insulin receptors, or due to defects in insulin receptor function, which has not been understood so far. The resistance to insulin response causes insulin to fail to initiate glucose uptake, oxidation, and storage in muscle tissue, while not effectively inhibiting lipolysis of adipose tissue and regulating glucose production and secretion in the liver.

Dipeptidyl peptidase-IV (DPP IV) is a serine protease that cleaves N-terminal dipeptides in a peptide chain containing a proline residue at the secondary end, despite the physiology of DPP IV in mammals. The role has not been fully confirmed, but it plays an important role in neuroenzyme metabolism, T-cell initiation, adhesion of cancer cells to endothelial tissues, and the entry of HIV into lymphoid cells (WO 98/19998).

Recently, studies have shown that DPP IV can block the secretion of glucagon-like peptide (GLP)-1, specifically, it can cleave the N-terminal histidine-alanine dipeptide from GLP-1. the active form of GLP-1 (7-36) NH ₂ into inactive degradation of _{GLP-1 (9-36) NH 2} (Endocrinology, 1999,140: 5356 ~ 5363). Due to physiological conditions, the half-life of intact GLP-1 in circulating blood is very short, and the inactive metabolites after DPP IV degrades GLP-1 can bind to GLP-1 receptor antagonistic activity GLP-1, thereby shortening the GLP-1. Physiological response. DPP IV inhibitors can completely protect endogenous and even exogenous GLP-1 from DPP IV inactivation, thereby greatly increasing the physiological activity of GLP-1 (5 to 10 fold), due to GLP-1 on pancreatic insulin Secretion is an important stimulator and directly affects glucose utilization, so DPP IV inhibitors are ideal for the treatment of non-insulin dependent diabetes mellitus (NIDDM) (US6110949).

However, although several DPP IV inhibitors have been disclosed, there are currently no long-acting drugs and there is still a need for DPP IV inhibitors with improved properties.

It is an object of the present invention to provide a compound having a therapeutic or palliative drug which inhibits DPP IV activity and which can be used for diabetes or the like.

The application PCT/CN2008/001936 filed on November 27, 2008 by the applicant of the present application describes a new class of tetrahydroimidazo[1,5-a]pyrene. Derivatives, and their use as DPP IV inhibitors, wherein Example 10 disclosed is ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butenyl] -3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyridyl The 1-formate form, which has been shown to have a significant inhibitory effect on DPP IV, is hereby incorporated by reference in its entirety.

Another object of the present invention is to provide pharmaceutically acceptable salt forms and compositions of the compounds of formula (I), thereby improving their solubility, bioavailability, hypoglycemic activity and pharmacokinetic properties.

The technical problem to be solved by the present invention is to provide novel ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butenyl]-3-trifluoromethyl-5. 6,7,8-tetrahydroimidazo[1,5-a]pyridyl A pharmaceutically acceptable salt of 1-carboxylic acid, a process for the preparation thereof, a pharmaceutical composition comprising the pharmaceutically acceptable salt, and the use thereof as a therapeutic agent, in particular as a dipeptidyl peptidase IV inhibitor. The salt-forming form has excellent activity for treating diabetes, has markedly improved solubility, and has good activity and bioavailability in animals, and low toxicity, and is suitable for preparing a preparation for treating diabetes.

The invention provides a pharmaceutically acceptable salt of a compound of formula (I). In the present invention, the "pharmaceutically acceptable salt" means a pharmaceutically unacceptable acid addition salt and a base addition salt. The acid addition salt is a salt of the compound of the formula (I) with a suitable inorganic or organic acid, including hydrochloride, phosphate, hydrogen phosphate, sulfate, hydrogen sulfate, sulfite. , acetate, oxalate, malonate, valerate, glutamate, oleate, palmitate, stearate, laurate, borate, p-toluenesulfonate, methane Acid salts, malate salts, tartrate salts, benzoates, pamoate, salicylates, vanillates, mandelates, succinates, gluconates, lactobions and lauryl sulfonic acids Salt, etc., especially phosphate. The base addition salt is a salt of a compound of the formula (I) with a suitable inorganic or organic base, and includes, for example, a salt with an alkali metal, an amine or a quaternary ammonium compound, such as a sodium salt or a lithium salt. Potassium salts, calcium salts, magnesium salts, amine salts, tetramethyl quaternary ammonium salts, tetraethyl quaternary ammonium salts, choline salts, especially choline salts. Amine salts, including salts with amines (NH ₃ ), primary amines, secondary amines or tertiary amines, such as methylamine salts, dimethylamine salts, trimethylamine salts, triethylamine salts, ethylamine salts, ethanolamine salts , a persalt and arginine, especially an ethanolamine salt.

Typical pharmaceutically acceptable salts of the compounds of formula (I) of the present invention include, but are not limited to:

Usually, the above preparation process can be carried out under cooling, normal temperature or heating conditions. It is noted that the selection of the reaction temperature has a certain influence on different salt formation reactions, which is also well known to those skilled in the art, and the salt formation reaction of the present invention. The temperature is from room temperature to the boiling point of the solvent used, preferably from 0 to 40 ° C; those skilled in the art can readily determine the optimum reaction temperature for a specific salt forming reaction by means of techniques conventional in the art.

The present invention relates to a process for the preparation of a pharmaceutically acceptable salt of a compound of formula (I), which comprises an acid addition salt process and a base addition salt process. The method for preparing an acid addition salt comprises ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butenyl]-3-trifluoromethyl-5,6,7 , 8-tetrahydroimidazo[1,5-a]pyridin The 1-formate hydrochloride is reacted with an alkaline solution, followed by the obtained ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3 -trifluoromethyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyridin 1-carboxylic acid is reacted with an inorganic or organic acid selected from the group consisting of hydrochloric acid, phosphoric acid, sulfuric acid, sulfurous acid, acetic acid, oxalic acid, malonic acid, valeric acid, glutamic acid, oleic acid , palmitic acid, stearic acid, lauric acid, boric acid, p-toluenesulfonic acid, methanesulfonic acid, malic acid, tartaric acid, benzoic acid, pamoic acid, salicylic acid, vanillic acid, mandelic acid, succinic acid, gluconic acid , lactobionic acid or lauryl sulfonic acid. The base addition salt preparation method comprises ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7, 8-tetrahydroimidazo[1,5-a]pyridin 1-carboxylic acid is reacted with an alkali metal hydroxide, a substituted amine or a quaternary ammonium compound, wherein the alkali metal hydroxide is selected from the group consisting of sodium hydroxide, lithium hydroxide, potassium hydroxide, and hydroxide. Calcium, magnesium hydroxide, the amine or quaternary ammonium compound selected from the group consisting of tetramethyl quaternary ammonium, tetraethyl quaternary ammonium, ethanolamine, choline, lysine, arginine, methylamine, two Methylamine, trimethylamine, triethylamine or ethylamine.

The present invention relates to the use of a pharmaceutically acceptable salt of a compound of formula (I) for the manufacture of a medicament for the treatment of type 2 diabetes, hyperglycemia, obesity or insulin resistance.

The invention relates to the use of a pharmaceutically acceptable salt of a compound of formula (I) for the preparation of a DPP IV inhibitor.

The present invention relates to a method of treating type II diabetes, hyperglycemia, obesity or insulin resistance comprising administering to a patient in need of treatment a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of formula (I).

The present invention relates to a method for inhibiting the catalytic activity of dipeptidyl peptidase IV, which comprises contacting the dipeptidyl peptidase IV with a pharmaceutically acceptable salt of a compound of formula (I).

The present invention relates to a pharmaceutically acceptable salt of a compound of formula (I) as a medicament for the treatment of type 2 diabetes, hyperglycemia, obesity or insulin resistance.

The present invention relates to a pharmaceutically acceptable salt of a compound of formula (I) as a medicament for inhibiting dipeptidyl peptidase IV.

The present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of formula (I), and a pharmaceutically acceptable carrier, and to a pharmaceutical composition for the treatment of type 2 diabetes, Use of drugs for hyperglycemia, obesity or insulin resistance.

The present invention relates to a method of treating type II diabetes, hyperglycemia, obesity or insulin resistance comprising administering to a patient in need of treatment a pharmaceutical composition comprising a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of formula (I).

The present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of formula (I) as a medicament for the treatment of type 2 diabetes, hyperglycemia, obesity or insulin resistance.

Unless otherwise stated, the following terms used in the specification and claims have the following meanings.

"Pharmaceutical composition" means a mixture of one or more pharmaceutically acceptable salts or prodrugs of a compound described herein with other chemical components, such as a physiological/pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to the organism.

[Synthesis method of the compound of the present invention]

In order to accomplish the object of the present invention, the present invention adopts the following technical solutions:

The method of synthesizing the compound of the formula (I) is prepared according to the method described in Example 10 of the application PCT/CN2008/001936 filed on Nov. 27, 2008, the disclosure of which is hereby incorporated by reference.

( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl The 1-carboxylic acid addition salt and the base addition salt are as follows:

Methods for acid addition salts include:

( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl The 1-formate hydrochloride is reacted with an alkaline solution, followed by the obtained ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3 -trifluoromethyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyridin 1-carboxylic acid is reacted with an inorganic or organic acid.

Methods for base addition salts include:

( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl The 1-carboxylic acid is reacted with an inorganic base or an organic base such as an alkali metal hydroxide, an amine or a quaternary ammonium salt in a water-miscible organic solvent.

detailed description

The following examples are intended to further illustrate the invention, but are not intended to limit the scope and spirit of the invention.

Example

The structure of the compound is determined by nuclear magnetic resonance ( ¹ H NMR) or mass spectrometry (MS). ^{The 1} H NMR shift (δ) is presented in parts per million (ppm). ^{The 1} H NMR measurement was performed on a Bruker AVANCE-400 nuclear magnetic apparatus, and the solvent was deuterated methanol (CD ₃ OD). The chemical shift was shown in units of 10 ^-6 (ppm); the measurement of MS was performed by FINNIGAN LCQAd (ESI) mass spectrometry. Instrument (manufacturer: Thermo, model: Finnigan LCQ advantage MAX); thin layer chromatography tantalum sheet using Yantai Yellow Sea HSGF254 or Qingdao GF254 tannin sheet, TLC used specifications are 0.15mm to 0.2mm, thin layer chromatography separation and purification products The specifications are from 0.4 mm to 0.5 mm.

Pipe column chromatography generally uses Yantai Huanghai Tanji 200 to 300 mesh gelatin as a carrier.

The starting materials of the present invention are known and commercially available from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Dari Chemicals, etc. The company may alternatively be synthesized or synthesized according to methods known in the art.

Unless otherwise specified in the examples, the reactions were all carried out under a nitrogen atmosphere; the nitrogen atmosphere means that the reaction flask was connected to a nitrogen balloon having a volume of about 1 L; and the hydrogen atmosphere means that the reaction flask was connected to a hydrogen balloon having a volume of about 1 L.

Unless otherwise stated in the examples, the solution means an aqueous solution.

There is no particular description in the examples, and the reaction temperature is room temperature.

The room temperature is an optimum reaction temperature of 20 ° C to 30 ° C.

The progress of the reaction in the examples was monitored by thin layer chromatography (TLC). The systems used for the reaction were: dichloromethane and methanol systems, n-hexane and ethyl acetate systems, petroleum ether and ethyl acetate systems. The volume ratio of acetone to solvent is adjusted depending on the polarity of the compound.

The column chromatography eluent system includes: A: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, the volume ratio of the solvent is adjusted according to the polarity of the compound, and may also be added. A small amount of amine water and acetic acid are adjusted.

HPLC refers to high performance liquid chromatography;

HPLC was measured using an Agilent 2695-2996 high pressure liquid chromatography (Gimini C18 150 x 4.6 mm column);

HPLC test conditions: Run time: 30 min Column temperature: 30 ° C PDA: 230 nm

Mobile phase: methanol: water (0.1% amine water) = 25: 75 flow rate: 1.0 mL / min

Example 1 (R)-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butenyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl -1-formate

first step

2,2-Dimethyl-5-[2-(2,4,5-trifluorophenyl)-ethinyl]-[1,3] Alkane-4,6-dione

2,2-dimethyl-[1,3] Alkyl-4,6-dione (5.69 g, 39.5 mmol) was dissolved in 400 mL of dichloromethane, and 2,4,5-trifluorophenylacetic acid 1a (7.15 g, 37.6 mmol) and p. Methylaminopyridine (7.35 g, 60.2 mmol), 250 mL of 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (8.28 g, 43.2 mmol) The methylene chloride suspension was stirred for 36 hours. The reaction mixture was washed with 5% aqueous potassium hydrogensulfate (250 mL×7) and brine (250 mL×2). 5-[2-(2,4,5-trifluorophenyl)-ethinyl]-[1,3] Alkane-4,6-dione 1b (11.4 g, white solid), yield: 96%.

MS m/z (ESI): 315.5 [M-1]

Second step 3-oxo-4-(2,4,5-trifluorophenyl)-butyric acid ethyl ester

2,2-Dimethyl-5-[2-(2,4,5-trifluorophenyl)-ethinyl]-[1,3] Alkane-4,6-dione 1b (15.72 g, 49.6 mmol) was dissolved in 280 mL of ethanol, and the reaction was stirred at 70 ° C for 12 hours. The mixture was cooled to room temperature, concentrated under reduced pressure, and the obtained residue was purified by eluent column chromatography using eluent system B to give 3-oxo-4-(2,4,5-trifluorophenyl)-butyric acid. Ethyl ester 1c (12 g, yellow liquid), yield: 88%.

MS m/z (ESI): 259 [M-1]

third step Ethyl 3-amino-4-(2,4,5-trifluorophenyl)-but-2-enoate

Ethyl 3-oxo-4-(2,4,5-trifluorophenyl)-butyrate 1c (24.6 g, 94.5 mmol) was dissolved in MeOH (MeOH) (EtOAc (EtOAc) The reaction was refluxed for 3 hours. The reaction mixture was concentrated under reduced pressure. EtOAc EtOAc. 50 mL of ethyl acetate was added to the obtained pale yellow solid, dissolved at 80 ° C, 50 mL of n-hexane was added, seeded, and cooled to room temperature. After 0.5 hours, 100 mL of n-hexane was added and allowed to stand in the refrigerator for 12 hours. Filtration gave ethyl 3-amino-4-(2,4,5-trifluorophenyl)-but-2-enoate 1d (19.5 g, white solid).

MS m/z (ESI): 260.1 [M+1]

the fourth step ( R )-3-t-butoxycarbonylamino-4-(2,4,5-trifluorophenyl)-butyric acid ethyl ester

Ethyl 3-oxo-4-(2,4,5-trifluorophenyl)-butyrate 1d (4.1 g, 15.8 mmol) was added to the autoclave, and 70 mL of methanol, dibutyl butyl dicarbonate was added. (3.8 g, 17.4 mmol), chloro(1,5-cyclooctadiene) ruthenium (I) dimer (32 mg, 0.063 mmol) and ( R )-1-[( S )-2-(diphenyl) Phosphyl)ferrocenyl]-ethyl-t-butylphosphine (68 mg, 0.13 mmol). The reaction was carried out for 24 hours at 6.67 atmospheres of hydrogen at 30 °C. Filtration, concentration of the filtrate under reduced pressure, adding 34 mL of methanol at 50 ° C, completely dissolving, adding 12 mL of water, cooling to room temperature, standing in the refrigerator for 12 hours, filtering, using methanol and water (V / V = 1 :1) Washing the solid product with a mixed solvent and drying in vacuo to give ( R )-3-t-butoxycarbonylamino-4-(2,4,5-trifluorophenyl)-butyric acid ethyl ester 1e (4 g , pale yellow solid), yield: 70%.

MS m/z (ESI): 362.4 [M+1]

the fifth step ( R )-3-Tertidinoxycarbonylamino-4-(2,4,5-trifluorophenyl)-butyric acid

Using a known method Tetrahedron Asymmetry , 2006, 17(2), 205-209, ( R )-3-t-butoxycarbonylamino-4-(2,4,5-trifluorophenyl)-butyric acid Ethyl ester 1e (10 g, 27.7 mmol) and sodium hydroxide (3.32 g, 83.1 mmol) were dissolved in 150 mL of a mixture solvent of methanol and water (V/V = 1:1). The reaction was stirred at 40 to 45 ° C for 1 to 1.5 hours, and concentrated under reduced pressure to remove a portion of solvent. After adding a small amount of water, 1 M hydrochloric acid was added dropwise to the reaction mixture to pH 2 to 3, and extracted with ethyl acetate (200 mL × 3). The organic phase was combined and washed with brine (200 mL) over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, ethyl acetate / n-hexane was recrystallized to give (R) -3- third butoxycarbonyl-amino-4- (2,4,5-trifluorophenyl) - Butyric acid 1f (9.2 g, white solid), used directly in the next step.

MS m/z (ESI): 332.3 [M-1]

Step 6 C-pyridyl -2-yl-methylamine

2-cyanopyridyl 1g (10.5 g, 100 mmol) dissolved in 150 mL 1,4-two In the alkane, 1.0 g of Raney nickel was added to a 250 mL autoclave, and the reaction was stirred at 40 ° C for 40 hours in 40 atmospheres of hydrogen. Filtration and concentration of the filtrate under reduced pressure gave C-pyrid -2-yl-methylamine 1 h (10.7 g, brown oil), yield: 98%.

MS m/z (ESI): I10 [M+1]

Seventh step 2,2,2-trifluoro-N-pyridyl -2-methyl-formamide

C-pyridyl 2-H-methylamine 1h (10.9 g, 100 mmol) was added to the reaction flask, and 20 mL of trifluoroacetic anhydride was added dropwise over 1 hour in an ice bath, and the reaction was stirred at room temperature for 2 hours, and the reaction solution was depressurized. Concentration, the residue obtained was purified by eluent column chromatography eluting to afford the title product 2,2,2-trifluoro-N-pyrid -2-Methyl-formamide 1i (21.0 g, brown oil).

MS m/z (ESI): 206.1 [M+1]

Eighth step 3-trifluoromethyl-imidazo[1,5-a]pyridinium

2,2,2-trifluoro-N-pyridyl 2-Methyl-carbamamine 1i (21.0 g, 100 mmol) was added to the reaction flask, and 100 mL of phosphorus oxychloride was added. After stirring for 30 minutes, phosphorus pentoxide (17.8 g, 125 mmol) was added. The reaction was refluxed for 5 hours and concentrated under reduced pressure. The reaction was quenched with deionized water, and a 20% sodium hydroxide solution was added dropwise to an aqueous solution to pH 5 to 6 and extracted with ethyl acetate (250 mL×4). Drying, filtration, and concentrating the filtrate under reduced pressure, and the residue obtained was purified by eluent column chromatography to elute to give the title product 3-trifluoromethyl-imidazo[1,5-a]pyridin. 1j (12.0 g, yellow solid), yield: 65%.

MS m/z (ESI): 188.0 [M+1]

¹ H NMR (400 MHz, CDCl ₃ , ppm): δ 9.15 (s, 1H), 8.06 (d, 1H), 7.92 (s, 1H), 7.81 (d, 1H)

Step 9 3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyridinium

3-trifluoromethyl-imidazo[1,5-a]pyridin 1j (12.0 g, 64.2 mmol) was dissolved in 150 mL of absolute ethanol and 500 mg of 10% palladium on carbon was added. The reaction was stirred for 12 hours under a hydrogen atmosphere. The reaction solution was filtered with EtOAc (EtOAc m.)jjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj 1k (12.2 g, brown solid), yield: 99%.

¹ H NMR (400 MHz, CDCl ₃ , ppm): δ 6.84 (s, 1H), 4.10 (m, 4H), 3.26 (m, 2H), 1.81 (s, 1H)

Step 10 ( R )-[3-oxo-1-(2,4,5-trifluorobenzyl)-3-(3-trifluoromethyl-5,6-dihydro-8H-imidazo[1,5 -a]pyridyl -7-yl)-propyl]-carbamic acid tert-butyl ester

Dissolve ( R )-3-t-butoxycarbonylamino-4-(2,4,5-trifluorophenyl)-butyric acid 1f (8.6 g, 45 mmol) and 9.4 mL of triethylamine in 300 mL After stirring for 5 minutes in dichloromethane, 3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyrene was added sequentially. 1k (15.0 g, 45 mmol) and bis (2-oxo-3- Azoliyl)phosphinium chloride (17.1 g, 67.3 mmol). The reaction was stirred for 2 hours, and the reaction liquid was concentrated under reduced pressure. The residue obtained was purified from the eluent column system B to obtain ( R )-[3-oxo-1-(2,4,5-three) Fluorobenzyl)-3-(3-trifluoromethyl-5,6-dihydro-8H-imidazo[1,5-a]pyridyl -7-yl)-propyl]-tert-butyl carbamate 1 m (20.0 g, white solid), yield: 88%.

¹ H NMR (400 MHz, CD ₃ OD, ppm): δ 7.25 (m, 1H), 7.11 (m, 1H), 7.032 (s, 1H), 4.93 (m, 2H), 4.35 (m, 3H) , 4.05 (m, 2H), 2.99 (m, 2H), 2.73 (m, 2H), 1.34 (s, 9H)

The eleventh step ( R )-[3-oxo-1-(2,4,5-trifluorobenzyl)-3-(1-bromo-3-trifluoromethyl-5,6-dihydro-8H-imidazole [1,5-a]pyridyl -7-yl)-propyl]-carbamic acid tert-butyl ester

( R )-[3-Oxo-1-(2,4,5-trifluorobenzyl)-3-(3-trifluoromethyl-5,6-dihydro-8H-imidazo[1, 5-a]pyridyl -7-yl)-propyl]-aminocarboxylic acid tert-butyl ester 1 m (20.0 g, 39.6 mmol) was dissolved in 300 mL of absolute ethanol, and N-brominated amber iodide (14.1 g, 79.2 mmol) was added. After stirring for 1 hour, potassium carbonate (10.9 g, 79.2 mmol) and di-tert-butyl dicarbonate (8.6 g, 39.6 mmol) were added and stirring was continued for 1 hour. The crude reaction mixture was filtered with silica gel, the filtrate was concentrated under reduced pressure, by silica gel column chromatography to elute System B was obtained residue was purified to give (R) - [3- oxo-1- (2,4,5 Trifluorobenzyl)-3-(1-bromo-3-trifluoromethyl-5,6-dihydro-8H-imidazo[1,5-a]pyridyl -7-yl)-propyl]-carbamic acid tert-butyl ester 1n (20.0 g, white solid), yield: 86%.

¹ H NMR (400 MHz, CDCl ₃ , ppm): δ 7.06 (m, 1H), 6.88 (m, 1H), 4.72 (s, 1H), 4.56 (s, 1H), 4.13 (m, 3H), 3.88 (m, 2H), 2.94 (m, 2H), 2.62 (m, 2H), 1.36 (s, 9H)

Step 12 ( R )-7-[3-Tertidinoxycarbonylamino-4-(2,4,5-trifluorophenyl)-butanindole-3-trifluoromethyl-5,6,7,8 -tetrahydroimidazo[1,5-a]pyridyl Methyl-1-carboxylate

Using a known method, Journal of Organometallic Chemistry , 1985, 285(1-3), 293-303, dicobalt octacarbonyl (4.02 g, 11.76 mmol), ethyl chloroacetate (0.71 g, 5.88 mmol), potassium carbonate ( 1.62 g, 11.76 mmol) and 50 mL of methanol were added to the reaction flask, and after stirring for 5 minutes, ( R )-[3-oxo-1-(2,4,5-trifluorobenzyl)-3-(1) was added. -Bromo-3-trifluoromethyl-5,6-dihydro-8H-imidazo[1,5-a]pyridin -7-yl)-propyl]-aminobutyl acid tert-butyl ester 1n (2.3 g, 3.92 mmol). The reaction mixture was stirred at 60 ° C for 2 hours, and the reaction mixture was concentrated under reduced pressure. The residue obtained was purified from the eluent column system B to obtain ( R )-7-[3-t-butoxycarbonylamino group. 4-(2,4,5-trifluorophenyl)-butenyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyridinium Methyl 1-benzoate 1p (1.1 g, white solid), yield: 50%.

MS m/z (ESI): 565.0 [M+1]

Step 13 ( R )-7-[3-Tertixocarbonylcarbonyl-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydro Imidazo[1,5-a]pyridyl 1-carboxylic acid

( R )-7-[3-Tertioxycarbonylamino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8 -tetrahydroimidazo[1,5-a]pyridyl Methyl 1-benzoate 1p (1.8 g, 3.2 mmol) was dissolved in 50 mL of methanol and 10 mL of 4 M sodium hydroxide solution was added. The reaction was stirred for 1 hour, and 2 M hydrochloric acid was added dropwise to the reaction mixture to pH 3 to 5, and the mixture was extracted with ethyl acetate (100 mL×3). Concentration under reduced pressure gave ( R )-7-[3-t-butoxycarbonyl-4-(2,4,5-trifluorophenyl)-butenyl]-3-trifluoromethyl-5,6,7 , 8-tetrahydroimidazo[1,5-a]pyridin 1-carboxylic acid 1q (1.76 g, pale yellow solid), yield: 100%.

MS m/z (ESI): 550.9 [M+1]

¹ H NMR (400 MHz, CD ₃ OD, ppm): δ 7.29-7.23 (m, 1H), 7.121-7.08 (m, 1H), 5.15-5.03 (m, 2H), 4.41-4.06 (m, 5H) ), 2.98-2.77 (m, 4H), 1.42-1.26 (m, 9H)

Fourteenth step ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl -1-formate

( R )-7-[3-Tertioxycarbonyl-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetra Hydrogen imidazo[1,5-a]pyridyl 1-carboxylic acid 1q (1.76 g, 3.2 mmol) was added to the reaction flask, and 10 mL of ethyl hydrogen chloride solution was added thereto, and the reaction was stirred for 1 hour, and the reaction mixture was concentrated under reduced pressure to give the title product ( R ) -7-[3-amine 4-(2,4,5-trifluorophenyl)-butenyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[1,5-a]pyridyl 1-formate hydrochloride 1 (1.56 g, white solid), yield: 100%.

MS m/z (ESI): 451.2 [M+1]

¹ H NMR (400 MHz, CD ₃ OD, ppm): δ 7.42-7.37 (m, 1H), 7.28-7.23 (m, 1H), 5.19-5.05 (m, 2H), 4.36-4.29 (m, 1H) ), 4.15-4.00 (m, 2H), 3.94-3.93 (m, 2H), 3.21-2.88 (m, 2H), 2.86-2.81 (m, 2H)

Example 2 (R)-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butenyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl 1-carboxylic acid phosphate

( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl 1-formate 1 (1.45 g, 2.97 mmol) was dissolved in 14 mL of dichloromethane, washed with 6 mL of saturated sodium bicarbonate, and the aqueous layer was extracted with 5.6 mL of dichloromethane. The mixture was washed with brine (6 mL), dried over anhydrous sodium sulfate , 2.97 mmol) of 2 mL of isopropanol solution, solid precipitated, stirred for 2 hours, filtered, and the filter cake was washed with cold isopropanol, and dried under reduced pressure at 40 ° C to give a crude product 1.44 g, yield 88.6%. The crude product (1.44 g, 2.63 mmol) was dissolved in 26 mL of isopropanol, stirred for 1 hour, filtered, and the filter cake was washed with isopropyl alcohol. The solid was dissolved in deionized water, concentrated under reduced pressure at 40 ° C, and at 40 ° C Drying in vacuo gave the title product ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8- Tetrahydroimidazo[1,5-a]pyridyl 1-carboxylic acid phosphate 2 (1.33 g, white powder), yield: 92.6%.

MS m/z (ESI): 451.2 [M+1]

¹ H NMR (400 MHz, CD ₃ OD, ppm): δ 7.36-7.42 (m, 1H), 7.19-7.25 (m, 1H), 5.01-5.15 (m, 2H), 4.24 - 4.34 (m, 2H) ), 4.06-4.11 (m, 1H), 3.91-3.98 (m, 1H), 3.07-3.12 (m, 2H), 2.8-3.09 (m, 2H)

Example 3 ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl 1-carboxylic acid

( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl 1-formate 1 (1.02 g, 2.1 mmol) was dissolved in 30 mL of methanol, sodium hydroxide solution (2.1 mL, 2.1 mmol) was added and the mixture was stirred for 15 min. The reaction mixture was concentrated under reduced pressure to give a solid and a mixed solvent of 15 mL of dichloromethane with methanol (V _{dichloromethane: methanol} = V 1: 3) was dissolved, filtered, and the filtrate was concentrated under reduced pressure to give the title product (R) -7 -[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[1,5-a Pyridine 1-carboxylic acid 3 (943 mg, white solid, HPLC: 99.89%), yield: 100%.

MS m/z (ESI): 451.2 [M+1]

¹ H NMR (400 MHz, CD ₃ OD, ppm): δ 7.32-7.41 (m, 1H), 7.11-7.21 (m, 1H), 5.00-5.07 (m, 2H), 4.16-4.24 (m, 2H) ), 4.05-4.08 (m, 1H), 3.85-3.97 (m, 2H), 3.05-3.17 (m, 2H), 2.91-2.93 (m, 2H)

Example 4 ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl Sodium 1-formate

(R)-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl 1-carboxylic acid 3 (100 mg, 0.22 mmol) was dissolved in 5 mL of methanol, sodium hydroxide solution (0.44 mL, 0.22 mmol) was added and the mixture was stirred for 15 min. The reaction mixture was concentrated under reduced pressure to give the title compound ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6, 7,8-tetrahydroimidazo[1,5-a]pyridyl Sodium 1-carboxylate 4 (104 mg, white solid, HPLC: 99.65%), yield: 99.7%.

MS m/z (ESI): 451.2 [M+1]

¹ H NMR (400 MHz, CD ₃ OD, ppm): δ 7.26-7.30 (m, 1H), 7.08-7.13 (m, 1H), 5.00-5.20 (m, 2H), 4.26-4.27 (m, 2H) ), 4.00-4.11 (m, 2H), 3.44 - 3.48 (m, 1H), 2.72 - 2.83 (m, 2H), 2.59 - 2.60 (m, 2H)

Example 5 ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl Lithium-1-carboxylate

( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl 1-carboxylic acid 3 (100 mg, 0.22 mmol) was dissolved in 5 mL of methanol, and a lithium hydroxide solution (0.44 mL, 0.22 mmol) was added and the mixture was stirred for 15 minutes. The reaction mixture was concentrated under reduced pressure to give the title compound ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6, 7,8-tetrahydroimidazo[1,5-a]pyridyl Lithium-1-carboxylate 5 (48 mg, white solid, HPLC: 99.66%), yield: 98.6%.

MS m/z (ESI): 451.2 [M+1]

¹ H NMR (400 MHz, CD ₃ OD, ppm): δ 7.31-7.38 (m, 1H), 7.13 - 7.22 (m, 1H), 5.07-5.27 (m, 2H), 4.26 - 4.36 (m, 2H) ), 4.01-4.15 (m, 2H), 3.53-3.60 (m, 1H), 2.80-2.91 (m, 2H), 2.59-2.72 (m, 2H)

Example 6 ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl Potassium-1-carboxylate

( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl 1-carboxylic acid 3 (100 mg, 0.22 mmol) was dissolved in 5 mL of methanol, then potassium hydroxide solution (0.44 mL, 0.22 mmol) was added and the mixture was stirred for 15 min. The reaction mixture was concentrated under reduced pressure to give the title compound ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6, 7,8-tetrahydroimidazo[1,5-a]pyridyl Potassium-1-carboxylate 6 (108 mg, white solid, HPLC: 92.78%), yield: 100%.

MS m/z (ESI): 451.2 [M+1]

¹ H NMR (400 MHz, CD ₃ OD, ppm): δ 7.31-7.38 (m, 1H), 7.14 - 7.23 (m, 1H), 5.07-5..27 (m, 2H), 4.26 - 4.36 ( m, 2H), 4.01-4.15 (m, 2H), 3.52-3.60 (m, 1H), 2.80-2.92 (m, 2H), 2.59-2.74 (m, 2H)

Example 7 ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl Calcium-1-carboxylate

( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl 1-carboxylic acid 3 (100 mg, 0.22 mmol) was dissolved in 10 mL of methanol. Calcium hydroxide (8.1 mg, 0.11 mmol) was added and the reaction was stirred for 20 hours. The reaction mixture was concentrated under reduced pressure to give the title compound ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6, 7,8-tetrahydroimidazo[1,5-a]pyridyl Calcium-1-carboxylate 7 (103 mg, white solid, HPLC: 99.60%), yield: 100%.

MS m/z (ESI): 451.2 [M+1]

¹ H NMR (400 MHz, CD ₃ OD, ppm): δ 7.28-7.34 (m, 1H), 7.11-7.21 (m, 1H), 5.10-5.21 (m, 2H), 4.22-4.36 (m, 2H) ), 4.03-4.09 (m, 2H), 3.55-3.59 (m, 1H), 2.76-2.85 (m, 2H), 2.60-2.71 (m, 2H)

Example 8 ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl 1-carboxylic acid triethylamine salt

( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl 1-carboxylic acid 3 (100 mg, 0.22 mmol) was dissolved in 5 mL of methanol, and a solution of triethylamine in methanol (0.767 mL, 0.22 mmol) was added, and 1 mL of triethylamine was added to methanol to prepare 25 mL. The reaction solution of triethylamine in methanol was stirred for 40 hours. The reaction mixture was concentrated under reduced pressure to give the title compound ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6, 7,8-tetrahydroimidazo[1,5-a]pyridyl 1-Acetic acid triethylamine salt 8 (112 mg, white solid, HPLC: 99.34%), yield: 99.8%.

MS m/z (ESI): 451.2 [M+1]

¹ H NMR (400 MHz, CD ₃ OD, ppm): δ 7.28-7.35 (m, 1H), 7.10-7.19 (m, 1H), 5.03-5.13 (m, 2H), 4.17 - 4.25 (m, 2H) ), 3.88-4.08 (m, 2H), 3.70-3.73 (m, 1H), 3.12-3.17 (m, 6H), 2.93-2.95 (m, 2H), 2.71-2.80 (m, 2H), 1.27-1.30 (m, 9H)

Example 9 ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl 1-carboxylic acid ethanolamine salt

( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl 1-carboxylic acid 3 (100 mg, 0.22 mmol) was dissolved in 5 mL of methanol, and ethanolic acid in methanol (0.33 mL, 0.22 mmol) was added. 1 mL ethanolamine was added to methanol to prepare 25 mL ethanolamine in methanol. ), the reaction was stirred for 40 hours. The reaction mixture was concentrated under reduced pressure to give the title compound ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6, 7,8-tetrahydroimidazo[1,5-a]pyridyl 1-carboxylic acid ethanolamine salt 9 (114 mg, white solid, HPLC: 99.62%), yield: 100%.

MS m/z (ESI): 451.2 [M+1]

^{] H NMR (400 MHz, CD} 3 OD, ppm): δ7.27-7.32 (m, 1H), 7.07-7.16 (m, 1H), 4.96-5.17 (m, 2H), 4.12-4.26 (m, 2H ), 3.91-4.09 (m, 2H), 3.70-3.72 (t, 2H), 3.56-3.57 (m, 1H), 2.95-2.98 (t, 2H), 2.80-2.89 (m, 2H), 2.58-2.70 (m, 2H)

Example 10 ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl -1-carboxylic acid choline salt

( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl 1-carboxylic acid 3 (100 mg, 0.22 mmol) was dissolved in 5 mL of methanol, and a solution of choline in methanol (1.55 mL, 0.22 mmol) was added. 1 mL of 45% choline was added to methanol to prepare 25 mL. The choline solution in methanol) was stirred for 3 hours. The reaction mixture was concentrated under reduced pressure to give the title compound ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6, 7,8-tetrahydroimidazo[1,5-a]pyridyl 1-C choline salt 10 (120 mg, white solid, HPLC: 99.41%), yield: 98.5%.

MS m/z (ESI): 451.2 [M+1]

¹ H NMR (400 MHz, CD ₃ OD, ppm): δ 7.23-7.30 (m, 1H), 7.06-7.15 (m, 1H), 4.99-5.19 (m, 2H), 4.19-4.26 (m, 2H) ), 3.89-4.07 (m, 4H), 3.60-3.71 (m, 1H), 3.50-3.55 (m, 2H), 3.21 (s, 9H), 2.72-2.84 (m, 2H), 2.55-2.66 (m , 2H)

Example 11 ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl 1-carboxylic acid malate

L-malic acid (368 mg, 2.74 mmol) was dissolved in 25 mL of a mixed solvent of methanol and water (V/V = 4:1) to prepare a 0.11 M solution. ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl 1-carboxylic acid 3 (100 mg, 0.22 mmol) was dissolved in 10 mL of methanol, 2 mL of the above-mentioned stock solution was added, and the reaction was stirred for 30 minutes. The reaction mixture was concentrated under reduced pressure to give the title compound ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6, 7,8-tetrahydroimidazo[1,5-a]pyridyl 1-carboxylic acid malate 11 (129 mg, white solid, HPLC: 98.92%), yield: 100%.

MS m/z (ESI): 451.1 [M+1]

¹ H NMR (400 MHz, CD ₃ OD, ppm): δ 7.32-7.41 (m, 1H), 7.16-7.23 (m, 1H), 4.96-5.13 (m, 2H), 4.35-4.39 (m, 1H) ), 4.20-4.30 (m, 2H), 4.04-4.13 (m, 1H), 3.90-4.00 (m, 2H), 3.07-3.13 (m, 2H), 2.77-2.97 (m, 3H), 2.56-2.62 (m, 1H)

Example 12 ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl -1-carboxylic acid tartrate

D-tartaric acid (413 mg, 2.75 mmol) was dissolved in 25 mL of a mixed solvent of methanol and water (V/V = 4:1) to prepare a 0.11 M solution. ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl 1-carboxylic acid 3 (100 mg, 0.22 mmol) was dissolved in 10 mL of methanol, 2 mL of the above-mentioned stock solution was added, and the reaction was stirred for 30 minutes. The reaction mixture was concentrated under reduced pressure to give the title compound ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6, 7,8-tetrahydroimidazo[1,5-a]pyridyl 1-carboxylic acid tartrate 12 (131 mg, white solid, HPLC: 99.35%), yield: 99%.

MS m/z (ESI): 451.1 [M+1]

¹ H NMR (400 MHz, CD ₃ 0D, ppm): δ 7.32-7.41 (m, 1H), 7.17-7.26 (m, 1H), 5.01-5.14 (m, 2H), 4.51 (s, 1H), 4.20-4.35 (m, 2H), 4.00-4.13 (m, 1H), 3.89-3.96 (m, 2H), 3.04-3.13 (m, 2H), 2.90-3.00 (m, 1H), 2.77-2.87 (m) , 1H)

Example 13 ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazo[ 1,5-a]pyridyl 1-carboxylic acid arginine

L-arginine (239 mg, 1.37 mmol) was dissolved in 25 mL of a mixed solvent of methanol and water (V/V = 4:1) to prepare a 0.055 M solution. ( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl 1-carboxylic acid 3 (100 mg, 0.22 mmol) was dissolved in 15 mL of methanol, 4 mL of the above-mentioned solution was added, and the reaction was stirred for 4 hours. The reaction mixture was concentrated under reduced pressure to give the title compound ( R )-7-[3-amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6, 7,8-tetrahydroimidazo[1,5-a]pyridyl 1-carboxylic acid arginine 13 (139 mg, white solid, HPLC: 98.89%), yield: 100%.

MS m/z (ESI): 451.1 [M+1]

¹ H NMR (400 MHz, CD ₃ OD, ppm): δ 7.24-7.32 (m, 1H), 7.09-7.14 (m, 1H), 4.98-5.18 (m, 2H), 4.25-4.28 (m, 1H) ), 4.18-4.19 (m, 1H), 3.93-4.04 (m, 2H), 3.50-3.54 (m, 2H), 3.18-3.23 (m, 2H), 2.76-2.87 (m, 2H), 2.58-2.69 (m, 2H), 1.77-1.90 (m, 2H), 1.67-1.75 (m, 2H)

Test case: Solubility test

The solubility of the compounds of the invention in four different systems was tested according to conventional solubility assays: phosphate buffer PBS (pH 7.4), methanol, 0.1% HCl, and water. The results are shown in Table 1:

Conclusion: The solubility of Example 2, Example 4 and Example 5 was significantly improved.

Biological evaluation

The following method can be used to determine the ability of the compounds of the invention to inhibit the activity of DPP IV, DPP VIII and DPP IX enzymes. The half-inhibitory concentration IC ₅₀ value of the compound (the concentration of the compound required to inhibit the enzymatic activity to 50%) is determined by reacting a certain amount of the enzyme with a substance and a different concentration of the test compound. Calculated.

By using the following test ^{Promega DPPIV-Glo TM Protease Assay (} Cat No. G8350 / G8351) inhibition assay kit of Example Compound DPP IV, DPP VIII and DPP IX activity embodiment of the present invention. among them:

a. DPP IV enzyme purchased from Calbiochem, Catalog no.317630

b. DPP VIII enzyme purchased from Bioscience, Catalog no.80080

c. DPP IX enzyme purchased from Bioscience, Catalog no.80090

For the detailed preparation process of the general reagents required for the assay (DPPIV-Glo buffer, luciferin reagents, etc.) and the detailed operation of the test, refer to the instructions of the above kit. The brief test method is as follows:

The test compound was dissolved in DMSO and formulated to the concentration required for the experiment. The DPP IV-Glo buffer and the lyophilized luciferin detection reagent were first equilibrated to room temperature, and then the fluorescein detection reagent powder was dissolved in a brown bottle to prepare a solution. The DPPIV-Glo substrate is then dissolved in ultrapure water to prepare the concentration required for the test, and then the substrate and the luciferase detection reagent solution (1:49 ratio used in this test) are thoroughly mixed in an appropriate ratio. And allowed to stand at room temperature for 30 to 60 minutes. A certain amount of Tris buffer (2 mM, pH 8.0), test compound and DPPIV (DPPVIII or DPPIX) enzyme were mixed uniformly and transferred to a 96-well reaction plate. Each test was set up with a duplicate well or a 3-well control, and the blank well and A negative volume of DMSO in the same volume as the test compound was added to the negative control well. Subsequently, a mixture of the prepared receptor-luciferase detecting reagent was added to each well to initiate the reaction, and the 96-well plate was sealed, and then incubated at room temperature for 40 minutes on a plate shaker. Then, the intensity of the fluorescent signal of each well was read by a microplate reader, and the inhibition rate of the compound at the concentration was calculated. The calculation formula is as follows:

Inhibition rate IR=[1-(S-B)/(N-B)]*100%

S: sample hole signal reading

B: blank control signal reading

N: negative control signal reading

IC ₅₀ values of the test compounds by inhibition rate can be calculated at different concentrations.

Conclusion: The free state and various salt compounds have good inhibitory activity against DPPPIV, and Example 2 has better selectivity.

Pharmacokinetic test Test Example 1 Pharmacokinetic test of the compound of the present invention 1. Purpose of the test

Rats were used as test animals, and the rats were administered with gavage and tail vein by LC/MS/MS method. Example 3 and gavage were given to Examples 1 to 4, Example 9, and Examples 11 to 12. The concentration of the drug in the plasma at the moment. The pharmacokinetic behavior of the compounds of the invention in rats was investigated, their pharmacokinetic profiles were evaluated, and their oral bioavailability was examined.

2, the test plan 2.1, test drugs

Compounds of Examples 1 to 4, compounds of Example 9, compounds of Examples 11 to 12

2.2, test animals

Twenty-eight healthy adult SD rats, male and female, were purchased from Shanghai Xipuer-Beikai Experimental Animal Co., Ltd., animal production license number: SCXK (Shanghai) 2008-0016.

2.3, equipment

API 4000 Q-trap Linear Ion Trap Mass Spectrometer, Applied Biosystems, USA; Agilent 1200 High Performance Liquid Chromatography System, Agilent, USA.

2.4, drug preparation

Intravenous group: Weigh the appropriate amount of the drug, add DMSO 0.5 mL ultrasonic solution, then dilute with physiological saline to 15 mL to make a 0.3 mg / mL solution.

The gavage administration group: Weighed the appropriate amount of the drug, and added 0.5% CMC-Na ultrasonic wave to make a 0.3 mg/mL suspension.

2.5, administration

Thirty-two healthy adult SD rats, half male and half female, were divided into 8 groups on average, with 4 rats in each group. After fasting overnight, Example 3 was administered by tail vein injection and Example 3 and its salt were administered by gavage at a dose of 3.0 mg/kg (in terms of alkali form) at a dose of 10 mL/kg.

2.6, sample collection

The intravenous administration group was administered with 0.2 mL of blood from the eyelids before administration and 2 minutes, 15 minutes, 30 minutes, 1.0 hours, 2.0 hours, 4.0 hours, 6.0 hours, 8.0 hours, 12.0 hours, and 24.0 hours after administration. In the heparinized test tube, the plasma was separated by centrifugation at 3500 rpm for 10 minutes, stored at -20 ° C, and fed 2 hours after administration.

The rats in the gavage administration group were subjected to blood collection before the administration and 0.5 hours, 1.0 hours, 2.0 hours, 3.0 hours, 4.0 hours, 6.0 hours, 8.0 hours, 12.0 hours, and 24.0 hours after the administration, and the sample treatment method was the same as the intravenous administration group. .

2.7, analytical methods

50 μL of rat plasma at each time after administration was added, 50 μL of internal standard solution and 150 μL of methanol were added, vortexed for 3 minutes, centrifuged for 10 minutes (13500 rpm), and 10 μL of the supernatant was taken for LC-MS/MS analysis.

2.8, pharmacokinetic parameter calculation

The pharmacokinetic behavior of the test compound was fitted to a compartment model, and the main pharmacokinetic parameters were calculated and calculated using DAS 2.0 software, in which C _max and t _{max were} measured. Oral absolute bioavailability was calculated according to AUC _0-t after intragastric and tail vein administration.

3, pharmacokinetic parameters results

The pharmacokinetic parameters of the compounds of the invention are shown in Table 2.

Conclusion: Compared with other compounds, Example 2 has significantly improved pharmacokinetic properties and bioavailability, and has obvious pharmacokinetic advantages.

Preliminary evaluation of hypoglycemic effect of the compounds of the invention First, the purpose of the test

The effects of the test compound Examples 1 to 4 and Examples 8 to 12 on the oral glucose tolerance of normal ICR mice (Shanghai Slack Laboratory Animal Co., Ltd.) were observed, and the mice were administered with glucose at different times within 2 hours. The sugar content in the tail blood was measured and analyzed to initially evaluate its hypoglycemic effect in the body.

Second, the test method 2.1 dose setting

The dose was 10 mg/kg, and the blank group was given water (both containing 5% DMSO).

2.2 method of administration

After intragastric administration, 10% glucose solution (0.8 mL per mouse) was administered at 4 g/kg 15 minutes after administration.

2.3 Determination of blood glucose levels

The blood glucose level (-15 minutes) was measured by dosing (Blank group was given water containing 5% DMSO).

After 15 minutes of administration, 20% glucose solution was administered at 4 g/kg, and blood glucose values of each mouse were measured at 0, 15, 30, 45, 60, and 120 minutes using a Roche Rocco whole blood glucose meter.

2.4 The test results are shown in Table 3:

Conclusion: Example 2 has significant hypoglycemic effects relative to other compounds.

Claims (9)

( R )-7-[3-Amino-4-(2,4,5-trifluorophenyl)-butanyl]-3-trifluoromethyl-5,6,7,8-tetrahydroimidazolium [1,5-a]pyridyl a salt of 1-carboxylic acid, wherein the salt is selected from the group consisting of a phosphate, a sodium salt, a lithium salt, or an ethanolamine salt, and the salt is as follows: The salt of claim 1, wherein the salt is a phosphate or an ethanolamine salt. A use of the salt according to claim 1 or 2 for the preparation of a medicament for the treatment of type 2 diabetes or hyperglycemia. A use of the salt of claim 1 or 2 for the preparation of a dipeptidyl peptidase IV inhibitor. The salt described in claim 1 or 2 is a drug for treating type 2 diabetes or hyperglycemia. The salt as described in claim 1 or 2 is a drug which inhibits dipeptidyl peptidase IV. A pharmaceutical composition comprising a therapeutically effective amount of the salt of claim 1 or 2 and a pharmaceutically acceptable carrier. A use of a pharmaceutical composition of claim 7 for the preparation of a medicament for the treatment of type 2 diabetes or hyperglycemia. The pharmaceutical composition of claim 7 is for use as a medicament for the treatment of type 2 diabetes or hyperglycemia.