A GLP-1R agonist, its synthesis method and uses thereof

Abstract

The present invention discloses a GLP-1R agonist, a synthesis method thereof and uses thereof, belonging to the field of medicine. The GLP-1R agonist of the present invention has a structure shown in formula (I) and can be used for treating GLP-1R-mediated diseases or disorders in humans, such as treating type 2 diabetes, prediabetes, obesity, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, and cardiovascular diseases.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and particularly relates to a GLP-1R agonist, a synthesis method thereof, and uses thereof. Background Art

[0002] Diabetes is a metabolic disease characterized by hyperglycemia. Due to factors such as insulin secretion defects or the physiological decline of pancreatic islet B cells, a long-term hyperglycemic state occurs in the body, which in turn leads to chronic damage or dysfunction of organs and tissues such as the eyes, kidneys, heart, blood vessels, and nerves. According to different pathogenesis, diabetes can be mainly divided into type I diabetes (T1D) and type II diabetes (T2DM), among which type II diabetes is caused by the decline of pancreatic islet B cell function and insulin resistance, and is the main type of diabetes.

[0003] Obesity is a chronic disease that is highly prevalent in modern society and is associated with many medical problems, including hypertension, hypercholesterolemia, and coronary heart disease. It is further highly correlated with T2DM and insulin resistance, which are usually accompanied by hyperinsulinemia or hyperglycemia or both. In addition, T2DM is associated with a 2- to 4-fold increased risk of coronary artery disease. Currently, bariatric surgery is the only treatment for efficiently eliminating obesity, but this treatment is expensive and risky. Pharmacological interventions are usually less effective and are associated with side effects.

[0004] Glucagon-like peptide-1 (GLP-1) is a peptide hormone secreted by enteroendocrine cells in the intestine in response to a meal. GLP-1 is thought to play a role in the regulation of postprandial blood glucose by directly increasing the secretion of insulin from pancreatic β cells induced by a meal and by promoting satiety by delaying the transit of food through the intestine. GLP-1 mediates intracellular signal transduction through the GLP-1 receptor (GLP-1R), which belongs to the family of G protein-coupled receptors present on the cell membrane and can lead to the accumulation of the second messenger cyclic adenosine monophosphate (cAMP) upon activation. Non-alcoholic steatohepatitis (NASH) can be associated with the features of metabolic syndrome, including obesity, type 2 diabetes, insulin resistance, and cardiovascular disease.

[0005] The early-developed GLP-1 receptor agonists are GLP-1 analogs, all of which are peptide substances, including liraglutide, semaglutide, etc. Through genetic engineering methods and formulation improvements, long-acting oral polypeptide drugs have been developed. However, oral polypeptides are still subject to various adverse limitations such as administration time, dose, and gastrointestinal disturbances. In recent years, researchers have found that the function of the GLP-1 receptor can also be regulated by small molecule compounds, and thus small molecule GLP-1 receptor agonists have been developed, including some low molecular weight flavonoid compounds, nitrogen-containing heterocyclic compounds, and compounds Boc-4 and S4P that mimic the polypeptide structure. However, the structural types of existing small molecule agonists are limited, and their specificity for the GLP-1 receptor is not strong, and their safety is not high. Therefore, it is necessary to develop agonists with new structural types and specific targeting of the GLP-1 receptor. Summary of the Invention

[0006] Problems to be Solved by the Invention

[0007] The present invention provides a GLP-1 receptor agonist having a 6-carboxyarylbenzimidazole structure, and this structural compound has better pharmacodynamic properties and higher safety.

[0008] Furthermore, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition, and uses.

[0009] Solutions for Solving the Problems

[0010] The present invention provides a compound of the structure shown in the following general formula (I) or a pharmaceutically acceptable salt thereof:

[0011]

[0012] Wherein:

[0013] A is selected from a heterocyclic group, a C3-7 cycloalkyl group, a phenyl group or a pyridyl group;

[0014] L is -(CH2)m-, where m is selected from 0, 1, 2, 3;

[0015] B is selected from a saturated heterocyclic group or a C3-7 cycloalkyl group;

[0016] X is selected from O, S, Se and CH2.

[0017] In one embodiment of the present invention, A is preferably selected from a piperidinyl group, a piperazinyl group, a cyclohexyl group, a phenyl group.

[0018] In one embodiment of the present invention, A is specifically selected from:

[0019] In one embodiment of the present invention, L is -(CH2)m-, preferably m is selected from 1 or 2;

[0020] In one embodiment of the present invention, when B is a saturated heterocyclic group, the heteroatom is selected from any one or a combination of N, O, S, and Se.

[0021] In one embodiment of the present invention, B is preferably selected from a piperidinyl group or a cyclohexyl group.

[0022] In one embodiment of the present invention, B is specifically selected from:

[0023] In one embodiment of the present invention, X is preferably selected from O or CH2;

[0024] In one embodiment of the present invention, the compounds of the structure shown in general formula (I) are specifically selected from:

[0025]

[0026] In one embodiment of the present invention, the pharmaceutically acceptable salt is an inorganic salt or an organic salt; the inorganic salts include hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, nitrate, phosphate, and acid phosphate; the organic salts are selected from acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, mesylate, tosylate, benzenesulfonate, and salicylate.

[0027] The present invention also provides the use of the compounds of the structure shown in the above general formula (I) or their pharmaceutically acceptable salts in the preparation of GLP-1 receptor agonists.

[0028] The present invention also provides a GLP-1 receptor agonist comprising the compounds of the structure shown in the above general formula (I) or their pharmaceutically acceptable salts.

[0029] The present invention also provides a pharmaceutical composition for treating GLP-1R-mediated diseases or disorders, which pharmaceutical composition comprises the compounds of the structure shown in the above general formula (I) or their pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier, excipient or diluent.

[0030] In another aspect, the invention encompasses the use of a therapeutically effective amount of a compound according to any of the embodiments for the manufacture of a medicament for treating an individual in need thereof suffering from cardiometabolic and related diseases, wherein said diseases are T1D, T2DM, prediabetes, idiopathic T1D (type 1b), LADA (latent autoimmune diabetes in adults), EOD (early-onset T2DM), YOAD (young-onset atypical diabetes), MODY (maturity-onset diabetes of the young), malnutrition-related diabetes, gestational diabetes, hyperglycemia, insulin resistance, hepatic insulin resistance, glucose intolerance, diabetic neuropathy, diabetic nephropathy, renal disease, diabetic retinopathy, adipocyte dysfunction, visceral fat accumulation, sleep apnea, obesity (including hypothalamic obesity and monogenic obesity) and related comorbidities (such as osteoarthritis and urinary incontinence), eating disorders (including binge eating syndrome, bulimia nervosa and syndromic obesity, such as Prader-Willi and Bardet-Biedl syndromes), weight gain caused by the use of other agents (such as the use of steroids and antipsychotics), sugar addiction, dyslipidemia (including hyperlipidemia, hypertriglyceridemia, increased total cholesterol, high LDL cholesterol and low HDL cholesterol), sugar addiction, dyslipidemia, hyperinsulinemia, NAFLD (including related diseases such as steatosis, NASH, fibrosis, cirrhosis and hepatocellular carcinoma), NASH, fibrosis, cirrhosis, hepatocellular carcinoma, cardiovascular disease, atherosclerosis (including coronary artery disease), coronary artery disease, peripheral vascular disease, hypertension, endothelial dysfunction, poor vascular compliance, congestive heart failure, myocardial infarction (such as necrosis and apoptosis), stroke, hemorrhagic stroke, ischemic stroke, traumatic brain injury, pulmonary hypertension, restenosis after angioplasty, intermittent claudication, postprandial dyslipidemia, metabolic acidosis, ketosis, arthritis, osteoporosis, Parkinson's disease, left ventricular hypertrophy, peripheral artery disease, macular lesions, cataracts, glomerulosclerosis, chronic renal failure, metabolic syndrome, syndrome X, premenstrual syndrome, angina, thrombosis, atherosclerosis, transient ischemic attack, vascular restenosis, glucose dysmetabolism, fasting glucose adverse symptoms, hyperuricemia, gout, erectile dysfunction, skin and connective tissue disorders, psoriasis, foot ulcers, ulcerative colitis, apo B hyperlipoproteinemia, Alzheimer's disease, schizophrenia, impaired cognitive function, inflammatory bowel disease, short bowel syndrome, Crohn's disease, colitis, irritable bowel syndrome, polycystic ovary syndrome for prevention or treatment and the treatment of addiction (such as alcohol and / or drug abuse).

[0031] The compounds of the present invention or their pharmaceutically acceptable salts can be formulated into solid preparations for oral administration, including, but not limited to, capsules, tablets, pills, powders, granules, etc. In these solid dosage forms, the compounds of general formula (I) of the present invention are mixed with at least one conventional inert excipient (or carrier) as the active ingredient, such as sodium citrate or calcium hydrogen phosphate. Or mixed with the following components: (1) fillers or solubilizers, such as starch, lactose, sucrose, glucose, mannitol, silicic acid, etc.; (2) binders, such as hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, gum arabic, etc.; (3) humectants, such as glycerol, etc.; (4) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates and sodium carbonate, etc.; (5) slow solvents, such as paraffin wax, etc.; (6) absorption accelerators, such as quaternary ammonium compounds, etc.; (7) wetting agents, such as cetyl alcohol and glycerol monostearate, etc.; (8) adsorbents, such as kaolin, etc.; (9) lubricants, such as talc, calcium stearate, solid polyethylene glycol, sodium lauryl sulfate, etc., or mixtures thereof. Capsules, tablets, and pills may also contain buffering agents.

[0032] The solid dosage forms such as tablets, dragees, capsules, pills, and granules can be coated with coating and shell materials such as enteric coatings and other polymorphic coatings or microencapsulation materials known in the art. They may contain opacifying agents, and the release of the active ingredient in such a composition can be delayed and released in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and wax substances. If necessary, the active ingredient can also be formed into microcapsule form with one or more of the above excipients.

[0033] The compounds of the present invention or their pharmaceutically acceptable salts can be formulated into liquid dosage forms for oral administration, including, but not limited to, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, tinctures, etc. In addition to the compounds of general formula (I) or their pharmaceutically acceptable salts as the active ingredient, the liquid dosage forms may contain inert diluents conventionally used in the art, such as water and other solvents, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, especially cottonseed oil, peanut oil, corn oil, olive oil, castor oil, sesame oil, etc., or mixtures of these substances. In addition to these inert diluents, the liquid dosage forms of the present invention may also include conventional adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances.

[0034] The suspending agents include, for example, ethoxylated octadecanol, polyoxyethylene sorbitol, and sorbitan, microcrystalline cellulose, agar, etc., or mixtures of these substances.

[0035] The compounds of the present invention and their pharmaceutically acceptable salts can be formulated into dosage forms for parenteral injection, including, but not limited to, physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions and dispersions. Suitable carriers, diluents, solvents, excipients include water, ethanol, polyols and their suitable mixtures.

[0036] The compounds of the present invention or their pharmaceutically acceptable salts can be formulated into dosage forms for topical administration, including, such as ointments, powders, suppositories, drops, sprays and inhalants, etc. The compound of general formula (I) of the present invention or its pharmaceutically acceptable salt as the active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and an optional preservative, buffer, and a propellant that may be required if necessary.

[0037] The pharmaceutical compositions of the present invention include the compound of general formula (I) or its pharmaceutically acceptable salt as the active ingredient, and a pharmaceutically acceptable carrier, excipient, diluent. When preparing the pharmaceutical composition, usually the compound of general formula (I) of the present invention or its pharmaceutically acceptable salt is mixed with a pharmaceutically acceptable carrier, excipient or diluent. The content of the compound of general formula (I) or its pharmaceutically acceptable salt can be 0.01 - 1000 mg, for example 0.05 - 800 mg, 0.1 - 500 mg, 0.01 - 300 mg, 0.01 - 200 mg, 0.05 - 150 mg, 0.05 - 50 mg, etc.

[0038] The beneficial effects of the present invention are as follows:

[0039] The present invention designs a class of compounds with a 6-carboxyarylbenzimidazole structure, which have a better biological function of activating GLP-1R and a relatively long metabolic half-life, enabling them to have the potential to reduce the medical dosage and extend the dosing time interval, thus providing a new means for finding new GLP-1R-mediated diseases or disorders. Detailed implementation manners

[0040] The present invention will be further described in detail below with reference to specific embodiments. The following embodiments are used to understand the method and core idea of the present invention. For those skilled in the art, any possible changes or substitutions made without departing from the concept of the present invention fall within the protection scope of the present invention. Unless otherwise specified, the raw materials and reagents used in the present invention are of chemical pure or higher purity.

[0041] Preparation of intermediates:

[0042] Synthetic route of 4-([1,4'-bipiperidin]-3-yloxy)methyl)-3-fluorobenzonitrile (5):

[0043]

[0044] Preparation of tert-butyl 3-((4-cyano-2-fluorobenzyl)oxy)piperidine-1-carboxylate (2):

[0045] Dissolve tert-butyl 3-hydroxypiperidine-1-carboxylate (9.38 g, 46.7 mmol) in DMF (100 mL). Add sodium hydride (2.24 g, 56.0 mmol) portionwise under ice bath. React at room temperature for 1 h. Slowly add a solution of 4-cyano-2-fluorobenzyl bromide (1) (10.00 g, 46.7 mmol) in DMF (20 mL) dropwise under ice bath and react at room temperature for 20 h. After the reaction is completed, pour the reaction solution into water and extract with ethyl acetate (200 mL × 3). Combine the organic phases, wash with water, saturated sodium chloride solution, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Purify the obtained crude product by silica gel column chromatography to obtain tert-butyl 3-((4-cyano-2-fluorobenzyl)oxy)piperidine-1-carboxylate (2) (12.00 g, 77%), a colorless oil. ESI-MS m / z: 335.1 [M+H] + 。

[0046] Preparation of 3-fluoro-4-((piperidin-3-yloxy)methyl)benzonitrile (3):

[0047] Dissolve tert-butyl 3-((4-cyano-2-fluorobenzyl)oxy)piperidine-1-carboxylate (2) (12.00 g, 35.9 mmol) in dichloromethane (100 mL), add trifluoroacetic acid (10 mL), stir at room temperature for 3 h. After the reaction is completed, concentrate under reduced pressure. Neutralize the residue with sodium bicarbonate, extract with dichloromethane (200 mL × 3), wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain 3-fluoro-4-((piperidin-3-yloxy)methyl)benzonitrile (3) (8.00 g, 95%), a colorless oil. ESI-MS m / z: 235.1 [M+H] + 。

[0048] Preparation of tert-butyl 3-((4-cyano-2-fluorobenzyl)oxy)-[1,4'-bipiperidine]-1'-carboxylate (4):

[0049] 3-Fluoro-4-((piperidin-3-yloxy)methyl)benzonitrile (3) (1.00 g, 4.3 mmol) and tert-butyl 4-oxopiperidine-1-carboxylate (0.85 g, 4.3 mmol) were dissolved in methanol (15 mL), 10% Pd-C (100 mg) was added, and the mixture was stirred at room temperature for 16 h under a hydrogen (1 atm) atmosphere. After completion of the reaction, the mixture was filtered and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography to give tert-butyl 3-((4-cyano-2-fluorobenzyl)oxy)-[1,4'-bipiperidine]-1'-carboxylate (4) (0.95 g, 53%), a white solid. ESI-MS m / z: 418.2 [M+H] + 。

[0050] Preparation of 4-([1,4'-bipiperidine]-3-yl-oxymethyl)-3-fluorobenzonitrile (5):

[0051] tert-Butyl 3-((4-cyano-2-fluorobenzyl)oxy)-[1,4'-bipiperidine]-1'-carboxylate (4) (0.95 g, 2.3 mmol) was dissolved in dichloromethane (15 mL), trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature for 3 h. After completion of the reaction, the mixture was concentrated under reduced pressure. The residue was neutralized with sodium bicarbonate, extracted with dichloromethane (50 mL×3), washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 4-([1,4'-bipiperidine]-3-yl-oxymethyl)-3-fluorobenzonitrile (5) (0.60 g, 83%), a white solid. ESI-MS m / z: 318.1 [M+H] + 。

[0052] Synthetic route of 3-fluoro-4-((3-(piperidin-4-yl)cyclohexyl)oxy)methyl)benzonitrile (13):

[0053]

[0054] Preparation of 1,4-dioxaspiro[4.5]decan-9-one (7):

[0055] 1,3-Cyclohexanedione (10.00 g, 89.28 mmol) and ethylene glycol (13.84 g, 223.20 mmol) were dissolved in toluene (100 mL), p-toluenesulfonic acid (200 mg) was added, and the mixture was heated to reflux for 16 h using a water separator. After completion of the reaction, the reaction mixture was washed with saturated sodium bicarbonate and brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 1,4-dioxaspiro[4.5]decan-9-one (7) (8.50 g, 61%), a white solid.

[0056] Preparation of 7-(pyridin-4-yl)-1,4-dioxaspiro[4.5]decan-7-ol (8):

[0057] 4 - Bromopyridine (8.00 g, 50.63 mmol) was dissolved in tetrahydrofuran (100 mL). Under nitrogen protection, isopropylmagnesium chloride (26.6 mL, 53.16 mmol, 2 M) was slowly added dropwise at 0 °C, and the mixture was stirred at 0 °C for 1 hour. Then, a solution of 1,4 - dioxaspiro[4.5]decan - 9 - one (7) (7.91 g, 50.63 mmol) in tetrahydrofuran (20 mL) was added dropwise. The mixture was allowed to warm to room temperature naturally and stirred for 16 hours. After the reaction was completed, it was quenched with saturated ammonium chloride and extracted with ethyl acetate (200 mL × 3). The combined organic phases were washed with water, saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography to obtain 7 - (pyridin - 4 - yl)-1,4 - dioxaspiro[4.5]dec - 7 - ol (8) (7.50 g, 62%), a white solid. ESI - MS m / z: 236.2 [M + H] +

[0058] Preparation of 4-(1,4 - dioxaspiro[4.5]dodec - 6 - en - 7 - yl)pyridine (9):

[0059] 7 - (pyridin - 4 - yl)-1,4 - dioxaspiro[4.5]dec - 7 - ol (8) (7.50 g, 31.91 mmol) was dissolved in pyridine (50 mL), thionyl chloride (10 mL) was added, and the mixture was heated to 100 °C and reacted for 1 hour. After the reaction was completed, it was cooled to room temperature and poured into ice - water (100 mL). It was neutralized with saturated sodium bicarbonate, extracted with dichloromethane (100 mL × 3), washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography to obtain 4-(1,4 - dioxaspiro[4.5]dodec - 6 - en - 7 - yl)pyridine (9) (5.15 g, 74%), a white solid. ESI - MS m / z: 218.1 [M + H] +

[0060] Preparation of 3-(piperidin - 4 - yl)cyclohexane - 1 - ol sulfate (10):

[0061] 4-(1,4 - dioxaspiro[4.5]dodec - 6 - en - 7 - yl)pyridine (9) (5.00 g, 23.04 mmol) was dissolved in ethanol (50 mL), platinum dioxide (500 mg) and concentrated sulfuric acid (2.26 g, 23.04 mmol) were added, and the mixture was stirred at room temperature for 16 hours under a hydrogen (1 atm) atmosphere. After the reaction was completed, it was filtered, and the filtrate was concentrated to obtain a crude product (6.00 g, 92%), a white solid.

[0062] Preparation of tert - butyl 4-(3 - hydroxycyclohexyl)piperidine - 1 - carboxylate (11):

[0063] 3-(Piperidin-4-yl)cyclohexan-1-ol sulfate (10) (6.00 g, 21.35 mmol) was dispersed in dichloromethane (100 mL). Triethylamine (9.0 mL, 64.05 mmol) and Boc anhydride (4.65 g, 21.35 mmol) were added successively. The mixture was stirred at room temperature for 16 h. After the reaction was completed, it was filtered. The filtrate was washed with water, dried over anhydrous sodium sulfate, and concentrated to obtain tert-butyl 4-(3-hydroxycyclohexyl)piperidine-1-carboxylate (11) (5.00 g, 82%), an anhydrous oil. ESI-MS m / z: 284.3 [M+H] + .

[0064] Preparation of tert-butyl 4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)piperidine-1-carboxylate (12):

[0065] The synthesis method was referred to the preparation of Intermediate 2. White solid, ESI-MS m / z: 417.1 [M+H] + .

[0066] Preparation of 3-fluoro-4-((3-(piperidin-4-yl)cyclohexyl)oxy)methyl)benzonitrile (13):

[0067] The synthesis method was referred to the preparation of Intermediate 3. White solid, ESI-MS m / z: 317.2 [M+H] + .

[0068] Synthetic route of 2-(4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)phenyl)acetic acid (20):

[0069]

[0070] Preparation of 3-oxocyclohex-1-en-1-yl trifluoromethanesulfonate (14):

[0071] 1,3-Cyclohexanedione (10.00 g, 89.28 mmol) was dissolved in dichloromethane (100 mL). Pyridine (14.11 g, 178.56 mmol) was added. The temperature was cooled to -78 °C, and trifluoromethanesulfonic anhydride (30.21 g, 107.14 mmol) was slowly added dropwise. The mixture was stirred at -78 °C for 10 min and then naturally warmed to room temperature. After the reaction was completed, 1 M dilute hydrochloric acid (200 mL) was added, and the mixture was extracted with dichloromethane (100 mL × 2). The organic phases were combined, washed with water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the obtained crude product was purified by silica gel column chromatography to obtain 3-oxocyclohex-1-en-1-yl trifluoromethanesulfonate (14) (15.50 g, 71%), a colorless oil.

[0072] Preparation of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-2-en-1-one (15):

[0073] Dissolve 3-oxocyclohex-1-en-1-yl trifluoromethanesulfonate (14) (10.00 g, 40.98 mmol) in 1,4-dioxane (100 mL), add bis(pinacolato)diboron (20.82 g, 81.96 mmol), potassium acetate (12.05 g, 122.94 mmol) and Pd(dppf)Cl2-DCM (1.66 g, 2.05 mmol), protect with nitrogen, heat to 80 °C and stir for 2 hours. After the reaction is completed, cool to room temperature, filter, concentrate the filtrate to obtain 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-2-en-1-one (15) (10.45 g), which is directly used in the next step without purification.

[0074] Preparation of tert-butyl 2-(5'-oxo-2',3',4',5'-tetrahydro-[1,1'-biphenyl]-4-yl)acetate (16):

[0075] Dissolve 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-2-en-1-one (15) (10.00 g, 45.04 mmol) and tert-butyl 2-(4-bromophenyl)acetate (10.17 g, 37.53 mmol) in 1,4-dioxane (150 mL), add 2M Na2CO3 (37.5 mL, 75.06 mmol) and tetrakis(triphenylphosphine)palladium(0) (2.16 g, 1.87 mmol), protect with nitrogen, heat to 100 °C and react for 16 hours. After the reaction is completed, cool to room temperature, filter, concentrate the filtrate, and purify the crude product by silica gel column to obtain tert-butyl 2-(5'-oxo-2',3',4',5'-tetrahydro-[1,1'-biphenyl]-4-yl)acetate (16) (6.00 g, 56%), a white solid.

[0076] Preparation of tert-butyl 2-(4-(3-oxocyclohexyl)phenyl)acetate (17):

[0077] Dissolve tert-butyl 2-(5'-oxo-2',3',4',5'-tetrahydro-[1,1'-biphenyl]-4-yl)acetate (16) (6.00 g, 20.98 mmol) in methanol (100 mL), add 10% Pd-C (500 mg), stir at room temperature for 16 hours under a hydrogen (40 psi) atmosphere. After the reaction is completed, filter, concentrate the filtrate under reduced pressure to obtain tert-butyl 2-(4-(3-oxocyclohexyl)phenyl)acetate (17) (5.85 g, 97%), a white solid.

[0078] Preparation of tert-butyl 2-(4-(3-hydroxycyclohexyl)phenyl)acetate (18):

[0079] Dissolve tert-butyl 2-(4-(3-oxocyclohexyl)phenyl)acetate (17) (5.00 g, 17.36 mmol) in methanol (50 mL), add sodium borohydride (0.65 g, 17.36 mmol) portionwise, stir at room temperature for 3 hours. After the reaction is completed, concentrate under reduced pressure, quench with saturated ammonium chloride, and extract with dichloromethane (100 mL × 2). Combine the organic phases, wash with water, wash with saturated sodium chloride, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and purify the obtained crude product by silica gel column to obtain tert-butyl 2-(4-(3-hydroxycyclohexyl)phenyl)acetate (18) (4.05 g, 80%), white solid, ESI-MS m / z: 291.1 [M+H] + 。

[0080] Preparation of tert-butyl 2-(4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)phenyl)acetate (19):

[0081] The synthesis method refers to the preparation of Intermediate 2. White solid, ESI-MS m / z: 424.1 [M+H] + 。

[0082] Preparation of 2-(4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)phenyl)acetic acid (20):

[0083] Dissolve tert-butyl 2-(4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)phenyl)acetate (19) (1 g, 2.36 mmol) in dichloromethane (15 mL), add 1 mL of trifluoroacetic acid, stir at room temperature for 3 hours. After the reaction is completed, wash the reaction solution with water, dry the organic phase over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain 2-(4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)phenyl)acetic acid (20) (0.65 g, 74%), white solid, ESI-MS m / z: 366.2 [M-H] - 。

[0084] Synthetic route of 4-(2-([3,4'-bipiperidin]-1-yl)ethyl)-3-fluorobenzonitrile (31):

[0085]

[0086] Preparation of methyl 2-(4-cyano-2-fluorophenyl)acetate (27):

[0087] Methyl 2-(4-bromo-2-fluorophenyl)acetate (5.50 g, 22.27 mmol) and Zn(CN)2 (2.60 g, 22.27 mmol) were dissolved in DMF (30 mL), Pd(PPh3)4 (1.28 g, 1.11 mmol) was added, and the mixture was heated to 80 °C under nitrogen protection and reacted for 16 hours. After the reaction, it was cooled to room temperature, 100 mL of water was added, and the mixture was extracted with ethyl acetate (100 mL * 3). The organic phases were combined, washed with water, saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the obtained crude product was purified by silica gel column chromatography to obtain methyl 2-(4-cyano-2-fluorophenyl)acetate (27) (3.05 g, 71%), a pale yellow liquid.

[0088] Preparation of 3-fluoro-4-(2-hydroxyethyl)benzonitrile (28):

[0089] Methyl 2-(4-cyano-2-fluorophenyl)acetate (27) (3.00 g, 15.54 mmol) was dissolved in methanol (50 mL), and lithium borohydride (677 mg, 31.08 mmol) was added portionwise under ice bath. The mixture was stirred at room temperature overnight. After the reaction, it was quenched with water and extracted with ethyl acetate (100 mL * 3). The organic phases were combined, washed with water, saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the obtained crude product was purified by silica gel column chromatography to obtain 3-fluoro-4-(2-hydroxyethyl)benzonitrile (28) (1.80 g, 70%), a pale yellow liquid.

[0090] Preparation of 4-(2-bromoethyl)-3-fluorobenzonitrile (29):

[0091] 3-Fluoro-4-(2-hydroxyethyl)benzonitrile (28) (1.50 g, 9.09 mmol) was dissolved in dichloromethane (30 mL), and phosphorus tribromide (3.69 g, 13.63 mmol) was slowly added under ice bath. The mixture was reacted at 0 °C for 2 hours. After the reaction, it was quenched with water, and the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the obtained crude product was purified by silica gel column chromatography to obtain 4-(2-bromoethyl)-3-fluorobenzonitrile (29) (1.86 g, 90%), a pale yellow liquid.

[0092] Preparation of tert-butyl 5,6-dihydro-[3,4'-bipyridine]-1(2H)-carboxylate (22):

[0093] 4-Bromopyridine hydrochloride (21) (3 g, 15.42 mmol) and tert-butyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (4.76 g, 15.42 mmol) were dissolved in 50 mL of DME:H2O (2:1). Sodium carbonate (8.17 g, 77.10 mmol) and Pd(PPh3)4 (0.35 g, 0.31 mmol) were added, and the reaction was carried out under nitrogen protection. The reaction mixture was stirred in a sealed tube at 100 °C for 16 hours. After the reaction was completed, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography to obtain tert-butyl 5,6-dihydro-[3,4'-bipyridine]-1(2H)-carboxylate (22) (1.85 g, 73%), a pale yellow solid, ESI-MS m / z: 261.1 [M+H] + 。

[0094] Preparation of tert-butyl [3,4'-bipiperidine]-1-carboxylate (23):

[0095] tert-Butyl 5,6-dihydro-[3,4'-bipyridine]-1(2H)-carboxylate (22) (1.80 g, 6.92 mmol) was dissolved in acetic acid (20 mL), and PtO2 (200 mg) was added. The reaction mixture was stirred at room temperature under a H2 balloon pressure for 16 hours. After the reaction was completed by TLC, the mixture was stirred at room temperature for 16 hours under a hydrogen (1 atm) atmosphere. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain tert-butyl [3,4'-bipiperidine]-1-carboxylate (23) (1.60 g, 86%), an anhydrous oil, ESI-MS m / z: 269.2 [M+H] + 。Preparation of tert-butyl 1'-benzyl-[3,4'-bipiperidine]-1-carboxylate (24):

[0096] tert-Butyl [3,4'-bipiperidine]-1-carboxylate (23) (1.50 g, 5.60 mmol) was dissolved in dichloromethane (30 mL), triethylamine (0.8 mL, 5.60 mmol) was added, and benzyl chloride (0.71 mL, 5.60 mmol) was slowly added. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was washed with water, the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain tert-butyl 1'-benzyl-[3,4'-bipiperidine]-1-carboxylate (24) (1.50 g, 75%), a pale yellow solid, ESI-MS m / z: 359.2 [M+H] + 。

[0097] Preparation of 1'-benzyl-3,4'-bipiperidine (25):

[0098] Dissolve tert-butyl 1'-benzyl-[3,4'-bipiperidine]-1-carboxylate (24) (1.50 g, 4.19 mmol) in dichloromethane (20 mL), add 2 mL of trifluoroacetic acid, and stir at room temperature for 3 hours. After the reaction is completed, concentrate under reduced pressure. The residue is neutralized with sodium bicarbonate, extracted with dichloromethane (50 mL * 3), washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 1'-benzyl-3,4'-bipiperidine (25) (0.60 g, 60%), a pale yellow oil. ESI-MS m / z: 259.1 [M+H] + 。

[0099] Preparation of 4-(2-(1'-benzyl-[3,4'-bipiperidine]-1-yl)ethyl)-3-fluorobenzonitrile (30):

[0100] Dissolve 1'-benzyl-3,4'-bipiperidine (25) (0.50 g, 1.94 mmol) and 4-(2-bromoethyl)-3-fluorobenzonitrile (29) (0.44 g, 1.94 mmol) in dichloromethane (10 mL), add triethylamine (0.5 mL, 3.88 mmol), and stir at room temperature for 16 hours. After the reaction is completed, wash the reaction mixture with water, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and purify the obtained crude product by silica gel column chromatography to obtain 4-(2-(1'-benzyl-[3,4'-bipiperidine]-1-yl)ethyl)-3-fluorobenzonitrile (30) (0.60 g, 77%), a pale yellow oil. ESI-MS m / z: 406.1 [M+H] + 。

[0101] Preparation of 4-(2-([3,4'-bipiperidine]-1-yl)ethyl)-3-fluorobenzonitrile (31):

[0102] Dissolve 4-(2-(1'-benzyl-[3,4'-bipiperidine]-1-yl)ethyl)-3-fluorobenzonitrile (30) (0.60 g, 1.48 mmol) in methanol (10 mL), add 10% Pd-C (50 mg), and stir at room temperature for 16 hours under a hydrogen (1 atm) atmosphere. After the reaction is completed, filter, and concentrate the filtrate under reduced pressure to obtain 4-(2-([3,4'-bipiperidine]-1-yl)ethyl)-3-fluorobenzonitrile (31) (0.45 g, 96%), a pale yellow solid. ESI-MS m / z: 316.1 [M+H] + 。

[0103] Synthetic route of 2-(4-(1-(4-cyano-2-fluorophenylethyl)piperidin-3-yl)phenyl)acetic acid (36):

[0104]

[0105] Preparation of ethyl 2-(4-(pyridin-3-yl)phenyl)acetate (33):

[0106] Ethyl 2-(4-bromophenyl)acetate (32) (5.00 g, 20.57 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (6.32 g, 30.85 mmol) were dissolved in 1,4-dioxane (100 mL), 2M Na2CO3 (20.5 mL, 41.14 mmol) and tetrakis(triphenylphosphine)palladium(0) (1.18 g, 1.03 mmol) were added, protected by nitrogen, and heated to 100 °C for reaction for 16 hours. After the reaction was completed, it was cooled to room temperature, filtered, the filtrate was concentrated, and the crude product was purified by silica gel column to obtain ethyl 2-(4-(pyridin-3-yl)phenyl)acetate (33) (3.00 g, 60%), white solid. ESI-MS m / z: 242.2 [M+H] + 。

[0107] Preparation of ethyl 2-(4-(piperidin-3-yl)phenyl)acetate (34):

[0108] Ethyl 2-(4-(pyridin-3-yl)phenyl)acetate (33) (3.00 g, 1.48 mmol) was dissolved in methanol (50 mL) and concentrated hydrochloric acid (1.5 mL), platinum dioxide (300 mg) was added, and stirred at room temperature for 4 hours under a hydrogen (1 atm) atmosphere. After the reaction was completed, it was filtered, the filtrate was concentrated under reduced pressure, neutralized with saturated sodium bicarbonate, extracted with dichloromethane (100 mL * 3), the organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain ethyl 2-(4-(piperidin-3-yl)phenyl)acetate (34) (2.85 g, 93%), light yellow liquid. ESI-MS m / z: 248.1 [M+H] + 。

[0109] Preparation of ethyl 2-(4-(1-(4-cyano-2-fluorophenylethyl)piperidin-3-yl)phenyl)acetate (35):

[0110] The synthesis method refers to the preparation of intermediate 30. Light yellow solid. ESI-MS m / z: 395 [M+H] + 。

[0111] Preparation of 2-(4-(1-(4-cyano-2-fluorophenylethyl)piperidin-3-yl)phenyl)acetic acid (36):

[0112] Ethyl 2-(4-(1-(4-cyano-2-fluorophenylethyl)piperidin-3-yl)phenyl)acetate (35) (1.00 g, 2.53 mmol) was dissolved in tetrahydrofuran (15 mL), and a solution of lithium hydroxide (182 mg, 7.59 mmol) in water (5 mL) was added. The reaction mixture was stirred at room temperature for 3 hours. After the reaction was completed, the pH was adjusted to 5 - 6 with 1 M dilute hydrochloric acid, and the mixture was extracted with dichloromethane (100 mL × 3). The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 2-(4-(1-(4-cyano-2-fluorophenylethyl)piperidin-3-yl)phenyl)acetic acid (36) (0.75 g, 80%), a pale yellow solid. ESI-MS m / z: 367.0 [M+H] + 。

[0113] Synthetic route of 3-fluoro-4-((1-(piperidin-4-ylmethyl)piperidin-3-yl)oxy)methyl)benzonitrile (40):

[0114]

[0115] Preparation of tert-butyl 4-(3-hydroxypiperidin-1-yl)methyl)piperidine-2-carboxylate (38):

[0116] tert-Butyl 4-formylpiperidine-1-carboxylate (37) (2.00 g, 9.39 mmol) was dissolved in methanol (30 mL), and piperidin-3-ol (0.95 g, 9.39 mmol) and acetic acid (0.56 g, 9.39 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours, then sodium cyanoborohydride (0.59 g, 9.39 mmol) was added, and the mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain tert-butyl 4-(3-hydroxypiperidin-1-yl)methyl)piperidine-2-carboxylate (38) (2.10 g, 75%), a pale yellow oil. ESI-MS m / z: 299.1 [M+H] + 。

[0117] Preparation of tert-butyl 4-(3-((4-cyano-2-fluorobenzyl)oxy)piperidin-1-yl)methyl)piperidine-1-carboxylate (39):

[0118] The synthesis method was referred to the preparation of intermediate 2. Pale yellow solid. ESI-MS m / z: 432.1 [M+H] + 。

[0119] Preparation of 3-fluoro-4-((1-(piperidin-4-ylmethyl)piperidin-3-yl)oxy)methyl)benzonitrile (40)

[0120] The synthesis method refers to the preparation of intermediate 3. Pale yellow solid. ESI-MS m / z: 332.0 [M+H] + .

[0121] Synthetic route of 2-(4-((3-((4-cyano-2-fluorobenzyl)oxy)piperidin-1-yl)methyl)phenyl)acetic acid (45):

[0122] Preparation of ethyl 2-(4-(bromomethyl)phenyl)acetate (42):

[0123] Dissolve ethyl 2-(p-tolyl)acetate (2.00 g, 11.23 mmol) in carbon tetrachloride (20 mL), add NBS (2.00 g, 11.23 mmol) and AIBN (183 mg, 1.12 mmol), protect with nitrogen, heat to 80 °C and react for 2 hours. After the reaction, cool to room temperature, filter, concentrate the filtrate, and purify the crude product by silica gel column to obtain ethyl 2-(4-(bromomethyl)phenyl)acetate (42) (1.72 g, 60%), a pale yellow oil.

[0124] Preparation of ethyl 2-(4-((3-hydroxypiperidin-1-yl)methyl)phenyl)acetate (43):

[0125] Dissolve ethyl 2-(4-(bromomethyl)phenyl)acetate (42) (1.50 g, 5.83 mmol) and piperidin-3-ol (0.59 g, 5.83 mmol) in dichloromethane (30 mL), add triethylamine (1.6 mL, 11.66 mmol), stir at room temperature for 16 hours. After the reaction, wash the reaction mixture with water, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and purify the obtained crude product by silica gel column to obtain ethyl 2-(4-((3-hydroxypiperidin-1-yl)methyl)phenyl)acetate (43) (0.92 g, 57%), a pale yellow oil. ESI-MS m / z: 278.3 [M+H] + .

[0126] Preparation of ethyl 2-(4-((3-((4-cyano-2-fluorobenzyl)oxy)piperidin-1-yl)methyl)phenyl)acetate (44):

[0127] The synthesis method refers to the preparation of intermediate 2. Pale yellow solid. ESI-MS m / z: 411.2 [M+H] + .

[0128] Preparation of 2-(4-((3-((4-cyano-2-fluorobenzyl)oxy)piperidin-1-yl)methyl)phenyl)acetic acid (45):

[0129] The synthesis method refers to the preparation of intermediate 36. Pale yellow solid. ESI-MS m / z: 383.1 [M+H] + .

[0130] Synthetic route of 3-fluoro-4-(((1-(2-(piperidin-4-yl)ethyl)piperidin-3-yl)oxy)methyl)benzonitrile (49):

[0131] Preparation of tert-butyl 4-(2-(3-hydroxypiperidin-1-yl)ethyl)piperidine-2-carboxylate (47):

[0132] The synthesis method refers to the preparation of intermediate 38. Colorless oil. ESI-MS m / z: 313.1 [M+H] + .

[0133] Preparation of tert-butyl 4-(2-(3-((4-cyano-2-fluorobenzyl)oxy)piperidin-1-yl)ethyl)piperidine-1-carboxylate (48):

[0134] The synthesis method refers to the preparation of intermediate 2. Pale yellow solid. ESI-MS m / z: 446.0 [M+H] + .

[0135] Preparation of 3-fluoro-4-(((1-(2-(piperidin-4-yl)ethyl)piperidin-3-yl)oxy)methyl)benzonitrile (49):

[0136] The synthesis method refers to the preparation of intermediate 3. Pale yellow oil. ESI-MS m / z: 346.1 [M+H] + .

[0137] Synthetic route of 3-fluoro-4-(((1-(2-(piperazin-1-yl)ethyl)piperidin-3-yl)oxy)methyl)benzonitrile (54):

[0138] Preparation of tert-butyl 4-(2-oxoethyl)piperazine-1-carboxylate (51):

[0139] Under nitrogen protection at -78 °C, a solution of DMSO (3.08 mL, 43.5 mmol) in DCM (20 mL) was added dropwise to a solution of oxalyl chloride (1.7 mL, 20.01 mmol) in DCM (20 mL). Stir for 0.5 h, then add a solution of tert-butyl 4-(2-hydroxyethyl)piperazine-1-carboxylate (50) (2.00 g, 8.70 mmol) in DCM (20 mL). Stir at -78 °C for 1 h, after adding triethylamine (3.7 mL, 26.10 mmol), stir the reaction at -78 °C for 30 min, and then stir at room temperature for 16 h. After the reaction was completed, 100 mL of water was added to dilute the reaction, and the mixture was extracted with DCM. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain tert-butyl 4-(2-oxoethyl)piperazine-1-carboxylate (51) (1.80 g, 91%), a pale yellow oil. ESI-MS m / z: 229.1 [M+H] + 。

[0140] Preparation of tert-butyl 4-(2-(3-hydroxypiperidin-1-yl)ethyl)piperazine-1-carboxylate (52):

[0141] The synthesis method was referred to the preparation of intermediate 38. Colorless oil. ESI-MS m / z: 314.1 [M+H] + 。

[0142] Preparation of tert-butyl 4-(2-(3-((4-cyano-2-fluorobenzyl)oxy)piperidin-1-yl)ethyl)piperazine-1-carboxylate (53):

[0143] The synthesis method was referred to the preparation of intermediate 2. Pale yellow solid. ESI-MS m / z: 447.2 [M+H] + 。

[0144] Preparation of 3-fluoro-4-((1-(2-(piperazin-1-yl)ethyl)piperidin-3-yl)oxy)methyl)benzonitrile (54):

[0145] The synthesis method was referred to the preparation of intermediate 3. Pale yellow oil. ESI-MS m / z: 347.0 [M+H] + 。

[0146] Synthetic route of 4-(2-(3-((4-cyano-2-fluorobenzyl)oxy)piperidin-1-yl)ethyl)benzoic acid (61): Preparation of 2-(4-(2-ethoxy-2-oxoethyl)phenyl)acetic acid (56):

[0147] Diethyl 2,2'-(1,4-phenylene)diacetate (55) (2.00 g, 8.00 mmol) was dissolved in tetrahydrofuran (30 mL), and an aqueous solution of lithium hydroxide (192 mg, 8.00 mmol) in water (10 mL) was added. The reaction was stirred at room temperature for 3 hours. After the reaction was completed, the pH was adjusted to 2 - 3 with 1 M dilute hydrochloric acid, and the mixture was extracted with dichloromethane (100 mL × 3). The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography to obtain 2-(4-(2-ethoxy-2-oxoethyl)phenyl)acetic acid (56) (1.20 g, 67%), a white solid. ESI-MS m / z: 221.1 [M - H] -

[0148] Preparation of ethyl 2-(4-(2-hydroxyethyl)phenyl)acetate (57):

[0149] 2-(4-(2-ethoxy-2-oxoethyl)phenyl)acetic acid (56) (1.00 g, 4.50 mmol) was dissolved in tetrahydrofuran (15 mL). Under an ice bath, borane-tetrahydrofuran complex (9 mL, 9.00 mmol, 1 M) was added, and then the mixture was heated to reflux for 3 hours. After the reaction was completed, it was cooled to 0 °C, and methanol was slowly added dropwise to quench the reaction. Water (50 mL) was added, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography to obtain ethyl 2-(4-(2-hydroxyethyl)phenyl)acetate (57) (0.65 g, 70%), a white solid. ESI-MS m / z: 209.3 [M + H] +

[0150] Preparation of ethyl 2-(4-(2-bromoethyl)phenyl)acetate (58):

[0151] The synthesis method was referred to the preparation of intermediate 29. A pale yellow liquid. ESI-MS m / z: 272.2 [M + H] +

[0152] Preparation of ethyl 4-(2-(3-hydroxypiperidin-1-yl)ethyl)benzoate (59):

[0153] The synthesis method was referred to the preparation of intermediate 30. A pale yellow solid. ESI-MS m / z: 278.1 [M + H] +

[0154] Preparation of ethyl 4-(2-(3-((4-cyano-2-fluorobenzyl)oxy)piperidin-1-yl)ethyl)benzoate (60):

[0155] The synthesis method refers to the preparation of Intermediate 2. Pale yellow solid. ESI-MS m / z: 411.2 [M+H] + .

[0156] Preparation of 4-(2-(3-((4-cyano-2-fluorobenzyl)oxy)piperidin-1-yl)ethyl)benzoic acid (61):

[0157] The synthesis method refers to the preparation of Intermediate 36. Pale yellow solid. ESI-MS m / z: 383.1 [M+H] + .

[0158] Synthetic route of 3-fluoro-4-((3-(piperazin-1-yl)cyclohexyl)oxy)methyl)benzonitrile (65):

[0159]

[0160] Preparation of tert-butyl 4-(3-hydroxycyclohexyl)piperazine-1-carboxylate (63):

[0161] Dissolve bis(2-chloroethyl)carbamic acid tert-butyl ester (62) (5.00 g, 20.66 mmol) and 3-aminocyclohexan-1-ol (2.37 g, 20.66 mmol) in acetonitrile (70 mL), add potassium iodide (1 g) and potassium carbonate (8.55 g, 61.98 mmol), and reflux under heating for 3 hours. After the reaction is completed, cool to room temperature, filter, concentrate the filtrate, and purify the obtained crude product through a silica gel column to obtain tert-butyl 4-(3-hydroxycyclohexyl)piperazine-1-carboxylate (63) (1.50 g, 24%), a pale yellow liquid. ESI-MS m / z: 285.2 [M+H] +

[0162] Preparation of tert-butyl 4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)piperazine-1-carboxylate (64):

[0163] The synthesis method refers to the preparation of Intermediate 2. Pale yellow solid. ESI-MS m / z: 418.2 [M+H] +

[0164] Preparation of 3-fluoro-4-((3-(piperazin-1-yl)cyclohexyl)oxy)methyl)benzonitrile (65)

[0165] The synthesis method refers to the preparation of Intermediate 3. Pale yellow oil. ESI-MS m / z: 318.2 [M+H] +

[0166] Synthetic route of 2-(1-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)piperidin-4-yl)acetic acid (70):

[0167]

[0168] Preparation of 3-fluoro-4-(((3-hydroxycyclohexyl)oxy)methyl)benzonitrile (67):

[0169] The synthesis method refers to the preparation of Intermediate 2. Pale yellow solid. ESI-MS m / z: 250.1 [M+H] +

[0170] Preparation of 3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl methanesulfonate (68):

[0171] Dissolve 3-fluoro-4-(((3-hydroxycyclohexyl)oxy)methyl)benzonitrile (67) (1.00 g, 4.01 mmol) in dichloromethane (15 mL), add triethylamine (1.1 mL, 8.02 mmol), and dropwise add methanesulfonyl chloride (551 mg, 4.81 mmol) under ice bath. After addition, stir at room temperature for 1 hour. After the reaction is completed, add water, extract with dichloromethane (10 mL×2), combine the organic phases, wash with brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain 3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl methanesulfonate (68) (1.10 g, 84%), a pale yellow liquid.

[0172] Preparation of ethyl 2-(1-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)piperidin-4-yl)acetate (69):

[0173] Dissolve 3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl methanesulfonate (68) (1.00 g, 3.06 mmol) in acetonitrile (15 mL), add potassium carbonate (0.84 g, 6.12 mmol), add ethyl 2-(piperidin-4-yl)acetate (0.52 g, 3.06 mmol) under ice bath, stir at room temperature for 6 hours. After the reaction is completed, quench with water. Extract with ethyl acetate (50 mL×3), combine the organic phases, wash with brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Purify the obtained crude product by silica gel column to obtain ethyl 2-(1-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)piperidin-4-yl)acetate (69) (0.52 g, 42%), an off-white solid.

[0174] Preparation of 2-(1-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)piperidin-4-yl)acetic acid (70):

[0175] The synthesis method refers to the preparation of Intermediate 36. Pale yellow solid. ESI-MS m / z: 375.2 [M+H] + .

[0176] Synthetic method of 2-(1'-(4-cyano-2-fluorophenethyl)-[1,3'-bipiperidin]-4-yl)acetic acid (74):

[0177]

[0178] The synthetic method refers to the preparation of intermediate 70. Pale yellow solid. ESI-MS m / z: 374.2 [M+H] + 。

[0179] (S)-tert-Butyl 2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (78) synthetic method:

[0180]

[0181] (S)-tert-Butyl 4-nitro-3-((oxetan-2-ylmethyl)amino)benzoate (76) preparation:

[0182] Dissolve tert-butyl 3-fluoro-4-nitrobenzoate (75) (5.00 g, 20.74 mmol) in acetonitrile (100 mL), add triethylamine (4.4 mL, 31.12 mmol) and (S)-oxetan-2-ylmethanamine (2.16 g, 24.88 mmol). Stir at room temperature for 16 h under nitrogen protection. After the reaction is completed, extract with ethyl acetate (100 mL×2), combine the organic phases, wash with brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain (S)-tert-butyl 4-nitro-3-((oxetan-2-ylmethyl)amino)benzoate (76) (5.50 g, 86%), yellow liquid, ESI-MS m / z: 309.1 [M+H] + , directly used for the next step.

[0183] (S)-tert-Butyl 4-amino-3-((oxetan-2-ylmethyl)amino)benzoate (77) preparation:

[0184] (S)-tert-Butyl 4-nitro-3-((oxetan-2-ylmethyl)amino)benzoate (76) (5.00 g, 16.23 mmol) was dissolved in ethanol (50 mL) and water (15 mL). Ammonium chloride (6.08 g, 113.61 mmol) and iron powder (4.54 g, 81.15 mmol) were added. The mixture was heated to 80 °C and reacted for 1 hour. After the reaction was completed, it was filtered while hot. The filtrate was extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography to obtain (S)-tert-butyl 4-amino-3-((oxetan-2-ylmethyl)amino)benzoate (77) (3.50 g, 77%), a yellow liquid, ESI-MS m / z: 279.2 [M+H] + 。

[0185] (S)-tert-Butyl 2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (78) Preparation:

[0186] (S)-tert-Butyl 4-amino-3-((oxetan-2-ylmethyl)amino)benzoate (77) (2.00 g, 7.19 mmol) was dissolved in acetonitrile (30 mL). 2-Chloro-1,1,1-trimethoxyethane (1.67 g, 10.79 mmol) and p-toluenesulfonic acid (50 mg) were added. The reaction solution was heated to 70 °C and reacted for 1 hour. After the reaction was completed, it was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography to obtain (S)-tert-butyl 2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (78) (1.50 g, 62%), a yellow liquid, ESI-MS m / z: 337.1 [M+H] + 。

[0187] Example 1: Synthesis of GLP-1R Agonist

[0188] Preparation of 2-((3-((4-cyano-2-fluorobenzyl)oxy)-[1,4'-bipiperidin]-1'-yl)methyl)-1-((S)-oxetan-2-yl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 1):

[0189]

[0190] Preparation of 2-((3-((4-cyano-2-fluorobenzyl)oxy)-[1,4'-bipiperidin]-1'-yl)methyl)-1-((S-oxetan-2-yl)methyl ester)-1H-benzo[d]imidazole-6-carboxylate (79):

[0191] (S)-tert-Butyl 2-(chloromethyl)-1-(oxan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (78) (100 mg, 0.30 mmol) and 4-(([1,4'-bipiperidin]-3-yl)oxymethyl)-3-fluorobenzonitrile (5) (95 mg, 0.30 mmol) were dissolved in acetonitrile (5 mL). Potassium carbonate (103 mg, 0.75 mmol) and potassium iodide (10 mg) were added. The reaction mixture was heated to 50 °C and reacted for 3 h. After the reaction was completed, water was added, and the mixture was extracted with ethyl acetate (200 mL × 3). The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography to give 2-((3-((4-cyano-2-fluorobenzyl)oxy)-[1,4'-bipiperidin]-1'-yl)methyl)-1-((S)-oxan-2-yl)methyl)-1H-benzo[d]imidazole-6-carboxylate (79) (120 mg, 65%), a yellow solid. ESI-MS m / z: 618.2 [M+H] + 。

[0192] Preparation of 2-((3-((4-cyano-2-fluorobenzyl)oxy)-[1,4'-bipiperidin]-1'-yl)methyl)-1-((S)-oxan-2-yl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 1):

[0193] 2-((3-((4-cyano-2-fluorobenzyl)oxy)-[1,4'-bipiperidin]-1'-yl)methyl)-1-((S)-oxan-2-yl)methyl)-1H-benzo[d]imidazole-6-carboxylate (79) (100 mg, 16.20 mmol) was dissolved in dichloromethane (2 mL). Trifluoroacetic acid (0.5 mL) was added, and the mixture was stirred at room temperature for 2 h. After the reaction was completed, it was concentrated under reduced pressure. The residue was separated by preparative HPLC to give 2-((3-((4-cyano-2-fluorobenzyl)oxy)-[1,4'-bipiperidin]-1'-yl)methyl)-1-((S)-oxan-2-yl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 1) (21.3 mg, 23%), a white solid. ESI-MS m / z: 562.1 [M+H] + 。

[0194] 11H NMR (400 MHz, DMSO-d6) δ: 12.15 (s, 1H), 8.51 (s, 1H), 8.12 (d, J = 8.8 Hz, 1H), 7.90 (d, J = 8.8 Hz, 1H), 7.56 - 7.22 (m 3H), 4.42 (s, 2H), 4.31 - 4.25 (m, 3H), 4.22 (s, 2H), 3.94 - 3.82 (m, 2H), 3.44 - 3.32 (m, 1H), 3.20 - 2.68 (m, 11H), 1.82 - 1.70 (m, 8H).

[0195] Using the corresponding intermediates or raw materials, the corresponding compounds shown in Table 1 were synthesized according to the operations similar to the basic operations for the preparation of Compound 1.

[0196] Table 1

[0197]

[0198] Preparation of 2-(4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)benzyl)-1-((S-oxetan-2-yl)methyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 3):

[0199]

[0200] Preparation of tert-butyl 4-(2-(4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)phenyl)acetamido)-3-(((S-oxetan-2-yl)methyl)amino)benzoate (80):

[0201] Dissolve (S)-tert-butyl 4-amino-3-((oxetan-2-ylmethyl)amino)benzoate (77) (150 mg, 0.54 mmol) and 2-(4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)phenyl)acetic acid (198 mg, 0.54 mmol) in DMF (2 mL), add DIPEA (188 μL, 1.08 mmol) and HATU (308 mg, 0.81 mmol), stir at room temperature for 3 hours. After the reaction is completed, add water, extract with ethyl acetate (20 mL * 3), combine the organic phases, wash with brine, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and purify the obtained crude product by silica gel column to obtain tert-butyl 4-(2-(4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)phenyl)acetamido)-3-(((S-oxetan-2-yl)methyl)amino)benzoate (80) (190 mg, 56%), yellow solid, ESI-MS m / z: 628.2 [M + H] +

[0202] Preparation of tert-butyl 2-(4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)benzyl)-1-((S-oxetan-2-yl)methyl)-1H-benzo[d]imidazole-6-carboxylate (81):

[0203] Dissolve tert-butyl 4-(2-(4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)phenyl)acetamido)-3-(((S-oxetan-2-yl)methyl)amino)benzoate (80) (150 mg, 0.24 mmol) in acetic acid (1 mL), heat to 60 °C and react for 3 hours. After the reaction is completed, add water, extract with ethyl acetate (20 mL × 3), combine the organic phases, wash with brine, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and purify the obtained crude product by silica gel column to obtain tert-butyl 2-(4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)benzyl)-1-((S-oxetan-2-yl)methyl)-1H-benzo[d]imidazole-6-carboxylate (81) (120 mg, 82%), a pale yellow solid, ESI-MS m / z: 610.2 [M+H] +

[0204] Preparation of 2-(4-(3-((4-cyano-2-fluorobenzyl)oxy)cyclohexyl)benzyl)-1-((S-oxetan-2-yl)methyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 3):

[0205] The synthesis method refers to the preparation of Compound 1, and Compound 3 is obtained by preparative HPLC separation. White solid, ESI-MS m / z: 554.1 [M+H] +

[0206] 1 1H NMR (400 MHz, DMSO-d6) δ: 12.13 (s, 1H), 8.42 (s, 1H), 8.11 (d, J = 8.8 Hz, 1H), 7.91 (d, J = 8.8 Hz, 1H), 7.56 - 7.21 (m 3H), 7.18 (d, J = 6.8 Hz, 2H), 7.12 (d, J = 6.8 Hz, 2H), 4.52 - 4.25 (m, 5H), 4.02 (s, 2H), 3.94 - 3.78 (m, 2H), 3.43 - 3.26 (m, 1H), 3.20 - 2.68 (m, 3H), 1.81 - 1.62 (m, 8H). Using the corresponding intermediates or raw materials, each compound shown in Table 2 is synthesized by an operation similar to the basic operation for the preparation of Compound 3.

[0207] Table 2

[0208]

[0209] Example 2: Biological Activity Assay of Compounds

[0210] The activity of the compounds of the present invention against GLP-1R was quantified by measuring the increase in cAMP in CHO cells (MultiSpan product #C1267-1a) stably expressing GLP-1R. Cells were harvested and plated at 1,000 cells / well in growth medium (DMEM / F-12 (Corning product #10-090-CV), supplemented with 10% FBS (HyClone product #SH30071-03), penicillin / streptomycin (Corning product #30-002CI) and 10 μg / ml puromycin (Gibco product #A11138-03)) in a 384-well plate (Greiner product #781080). The cells were then incubated overnight at 37 °C and 5% CO2. The next day, the medium was removed, the cells were washed with DPBS (Corning product #21-031-CM), and then assay medium (HBSS, Corning product #21-023-CV) containing 20 mM Hepes (Gibco product #15630-080) and 0.1% BSA (Rockland Immunochemicals product #BSA-1000) was added. After the medium change, the cells were incubated at 37 °C and 5% CO2 for 1 hour. The test GLP-1 compounds were added to the cells in a 10-point dose response, and then incubated at 37 °C and 5% CO2 for 30 minutes. The increase in cAMP concentration was detected using Cisbio's cAMPGs Dynamic Kit (product #62AM4PEC). The response was plotted against the logarithm of the agonist concentration and fitted to a sigmoidal equation to determine the EC50 value. The specific results are shown in Table 3.

[0211] Table 3 EC of Exemplary Compounds against GLP-1 Agonist Activity 50 Value (nM)

[0212] Compound <![CDATA[Activity (EC 50 )(nM)]]> PF-06882961 1.1 Compound 1 0.8 Compound 2 0.7 Compound 3 1.1 Compound 4 1.0 Compound 5 0.6 Compound 6 0.5 Compound 7 1.2 Compound 8 0.6 Compound 9 0.9 Compound 10 1.1 Compound 11 0.8 Compound 12 1.0 Compound 13 0.7

[0213] Example 3: Evaluation of Compound Stability Using Human Liver Microsomes

[0214] The hepatic microsomal enzyme stability of each compound was compared with PF-06882961.

[0215] Determination System: The metabolic stability of the compounds of the present invention was tested using a mixture of male and female liver microsomes with 1 mM NADPH. The samples were analyzed using a mass spectrometer. HRMS was used to determine the peak area response ratio (the peak area corresponding to the test compound or reference divided by the peak area of the analytical internal standard) without running a standard curve. To detect all possible metabolites, HRMS scans were performed over an appropriate m / z range.

[0216] Determination Conditions: The determination was carried out with a single incubation (N = 1). The test compound was incubated at 37 °C in a buffer containing 0.5 mg / mL liver microsomal protein. The reaction was initiated by adding the cofactor, and samples were taken at 0, 0.5, 1, 2, 4, 8, 16, 24 hours. A positive control (5 μM testosterone) was incubated in parallel and samples were taken at 0, 0.5, 1, 2, 4, 8, 16, 24 hours.

[0217] Determination Quality Control: The reference compound testosterone was run in parallel to confirm the enzyme activity of the (liver) microsomes. After the final time point, fluorescence assay was used to confirm the addition of NADPH to the reaction mixture. The T1 / 2 of the reference met the acceptable internal standard.

[0218] Analytical Method:

[0219] Liquid Chromatography Column: Thermo BDS Hypersil C18 30X2.0mm, 3μm, with a guard column M.P., Buffer: 25 mM formic acid in buffer, pH 3.5;

[0220] Aqueous Phase (A): 90% water, 10% buffer;

[0221] Organic Phase (B): 90% acetonitrile, 10% buffer;

[0222] Flow Rate: 300 μL / min;

[0223] Autosampler: Injection volume 10 μL;

[0224] The gradient program is shown in Table 4.

[0225] Table 4 Gradient Program

[0226] Time (minutes) ％A ％B 0.0 100 0 1.5 0 100 2.0 0 100 2.1 100 0 3.5 100 0

[0227] By using human liver microsomes, Examples 1, 2, 3, 4, 5, 11, 12, and 13 described in the present invention exhibited a metabolic half-life greater than 4 hours, and Examples 6, 7, 8, 9, and 10 exhibited a metabolic half-life between 2 and 4 hours, significantly greater than the 1.8-hour metabolic half-life of PF-06882961. The results showed that the introduction of low-polarity groups in the compounds was beneficial to reducing the lipophilicity of the compounds, resulting in a reduced probability of the compounds of the present invention being metabolized by metabolic enzymes, and the relatively long metabolic half-life of the compounds enabled them to have the potential to reduce the medical dosage and extend the dosing time interval.

[0228] For compounds of the general formula (I), the linking group and the substituent group have an important influence on the pharmacodynamic properties of the compounds.

[0229] Although the present invention has been illustrated by the previous specific examples, it should not be construed as being limited thereto; rather, the present invention encompasses the general aspects disclosed previously. Various modifications and various embodiments can be made without departing from the spirit and scope of the present invention.

Claims

1. A compound of the structure shown in general formula (I) or a pharmaceutically acceptable salt thereof: Wherein: A is selected from piperidinyl, piperazinyl, phenyl; L is -(CH2)m-, where m is selected from 0, 1, 2, 3; B is selected from piperidinyl or cyclohexyl; X is selected from O, CH2.

2. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein, The compounds of the structure shown in general formula (I) are specifically selected from:

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The pharmaceutically acceptable salts are inorganic salts or organic salts; the inorganic salts include hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, nitrate, phosphate, acid phosphate; the organic salts are selected from acetate, trifluoroacetate, propionate, pyruvate, glycolate, oxalate, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, mesylate, tosylate, benzenesulfonate, salicylate.

4. Use of the compound according to any one of claims 1-3 or a pharmaceutically acceptable salt thereof in the preparation of a GLP-1 receptor agonist.

5. A GLP-1 receptor agonist containing the compound according to any one of claims 1-3 or a pharmaceutically acceptable salt thereof.

6. A pharmaceutical composition for treating a GLP-1R-mediated disease or disorder, characterized in that, The pharmaceutical composition contains the compound according to any one of claims 1-3 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, excipient or diluent.

7. Use of the compound according to any one of claims 1-3 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating patients with cardiometabolic and related diseases.

8. The application according to claim 7, wherein Wherein the diseases are selected from type I diabetes, type II diabetes, prediabetes, idiopathic type I diabetes, latent autoimmune diabetes, maturity-onset diabetes of the young, early-onset diabetes, malnutrition-related diabetes, gestational diabetes, hyperglycemia, insulin resistance, hepatic insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, adipocyte dysfunction, visceral fat deposition, obesity, eating disorders, sleep apnea, weight gain, sugar craving, dyslipidemia, hyperinsulinemia, congestive heart failure, myocardial infarction, hemorrhagic stroke, ischemic stroke, traumatic brain injury, pulmonary hypertension, restenosis after angioplasty, intermittent claudication, postprandial lipemia, metabolic acidosis, ketosis, arthritis, left ventricular hypertrophy, Parkinson's disease, peripheral arterial disease, macular degeneration, cataract, glomerulosclerosis, chronic renal failure, colic, premenstrual syndrome, thrombosis, atherosclerosis, impaired glucose metabolism, restenosis or Alzheimer's disease.

9. The application according to claim 7, characterized in that, Wherein the diseases are selected from kidney diseases, metabolic syndrome, stroke, dementia.

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