Compounds as GLP-1 receptor agonists and their uses

Abstract

The application provides a compound as shown in formula (I) and use thereof. The compound as shown in formula (I) provided by the application can be used as an effective glucagon-like peptide-1 (GLP-1) receptor agonist, has excellent GLP-1R receptor agonist activity and obvious blood glucose lowering effect, and has good pharmacokinetic characteristics, thereby providing more choices for the prevention and / or treatment of diseases, conditions or disorders related to GLP-1 activity, and having a good application prospect in clinic.

Description

[0001] Cross-references

[0002] This application claims priority to the Chinese patent application filed on October 22, 2021, with application number 202111235046.8 and invention name “Compounds as GLP-1 receptor agonists and their uses”, the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present invention relates to the field of medicinal chemistry, and in particular to a compound serving as a GLP-1 receptor agonist and its use. Background Art

[0004] Diabetes has become one of the most important public health issues in today's society due to its increasing prevalence and associated health risks.

[0005] This disease is characterized by hyperglycemia caused by defects in insulin secretion and / or insulin action. Two main forms of diabetes have been identified: type 1 diabetes and type 2 diabetes. Type 1 diabetes (T1D) is an autoimmune disease targeting pancreatic beta cells, involving a variety of immune cells. Currently, the main treatment for T1D is insulin injection. Although this treatment can effectively control the patient's blood sugar level, it cannot prevent the progressive failure of pancreatic islet function. Type 2 diabetes (T2DM), also known as non-insulin-dependent diabetes, is caused by insulin resistance and insufficient insulin secretion.

[0006] A variety of pharmacological approaches are currently available for the treatment of hyperglycemia and, subsequently, T2DM. These approaches can be divided into six major categories: (A) insulin secretagogues, including sulfonylureas, meglitinides, dipeptidyl peptidase IV (DPP-IV) inhibitors, and glucagon-like peptide-1 receptor (GLP-1R) agonists; (B) biguanides, such as metformin; (C) α-glucosidase inhibitors, such as acarbose; (D) thiazolidinediones, such as pioglitazone; (E) insulin, alone or in combination with the above agents; and (F) sodium-glucose co-transporter 2 (SGLT2) inhibitors, such as dapagliflozin. With the exception of GLP-1 receptor agonists and SGLT2 inhibitors, all of these agents have limited efficacy and fail to address the most important issue currently under investigation: β-cell dysfunction and the associated obesity.

[0007] Obesity is a chronic disease that is highly prevalent in modern society and is associated with many medical problems, including hypertension, hypercholesterolemia, and cardiovascular and cerebrovascular diseases, in addition to being highly associated with T2DM and insulin resistance, which is often accompanied by hyperinsulinemia and / or hyperglycemia. Currently, bariatric surgery is one of the main therapeutic methods for treating obesity, but this treatment method is expensive and risky. Pharmacological intervention is usually less effective and has side effects. Therefore, it is clear that there is a need for a more effective pharmacological intervention method for obesity that has fewer side effects and is convenient to administer.

[0008] T2DM is associated with, in addition to hyperglycemia and insulin resistance, liver insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, obesity, dyslipidemia, hypertension, hyperinsulinemia, and nonalcoholic fatty liver disease (NAFLD).

[0009] Glucagon-like peptide-1 (GLP-1) is an incretin consisting of 30 or 31 amino acid residues that is secreted by the neuroendocrine L cells of the distal ileum and colon in response to food intake. GLP-1 has been shown to stimulate insulin secretion in a physiological and glucose-dependent manner, to decrease glucagon secretion, to suppress gastric emptying, to decrease appetite, and to stimulate beta cell proliferation. In T2DM patients, the normal postprandial rise of GLP-1 is missing or reduced (Vilsboll T. et al. Diabetes. 2001. 50; 609-613).

[0010] GLP-1 receptor agonists (e.g., exenatide, etc.) can improve glycemic control in T2DM patients by lowering fasting and postprandial glucose, and their main pharmacological activities include: (i) increasing glucose-dependent insulin secretion, (ii) glucagon-inhibitory activity in hyperglycemic conditions, (iii) delaying gastric emptying rate, resulting in delayed absorption of meal-derived glucose (see Holst (Physiol. Rev. 2007, 87, 1409) and Meier (Nat. Rev. Endocrinol. 2012, 8, 728)). Although these receptor agonists provide good therapeutic effects for patients, they still only account for a limited proportion of diabetes prescriptions, one of the important reasons being that they are all macromolecular peptide-based drugs that need to be administered by invasive subcutaneous injection, which has poor patient compliance relative to oral administration.

[0011] PF-06882961 is an oral small molecule GLP-1 receptor agonist developed by Pfizer. The results of the phase I clinical trial of PF-06882961 showed that it had a significant effect on glucose lowering and weight loss in patients with type 2 diabetes.

[0012] Although oral small molecule GLP-1 receptor agonist PF-06882961 has been reported, there is still a need to develop new oral small molecule compounds with better pharmacodynamic and pharmacokinetic properties for the prevention and / or treatment of diseases related to GLP-1 activity.

[0013] The information disclosed in this Background section is only for the purpose of increasing the understanding of the general background of the application and does not necessarily constitute an acknowledgement or any form of suggestion that this information forms the prior art already known to those of ordinary skill in the art. SUMMARY

[0014] The object of the present application is to provide a compound as a GLP-1 receptor agonist and its use. The compound has excellent GLP-1R receptor agonistic activity and a clear hypoglycemic effect, and has better pharmacokinetic characteristics, providing more options for the prevention and / or treatment of diseases, conditions or disorders related to GLP-1 activity.

[0015] In particular, in a first aspect, the present application provides a compound as represented by the general formula (I) or a pharmaceutically acceptable salt thereof:

[0016]

[0017] wherein,

[0018] R1is selected from (S)-oxetan-2-yl, 1-tetrahydrofuran-3-yl or 1-ethyl-1H-imidazol-4-yl or oxazol-2-yl;

[0019] R2is selected from hydrogen or halogen;

[0020] R3is selected from hydrogen, C1-C6alkyl or halogen;

[0021] R4is selected from hydrogen, optionally substituted C1-C6alkyl or optionally substituted C3-C7cycloalkyl; when R4is substituted C1-C6alkyl or substituted C3-C7cycloalkyl, the substituents can be at any available point of attachment, and the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, amino, haloalkyl, hydroxyalkyl, carboxyl or carboxylate;

[0022] R5is selected from hydrogen or halogen;

[0023] X, Z and Z1are each independently selected from -CH or nitrogen.

[0024] As a preferred embodiment, in the compound represented by the above formula (I),

[0025] R1is (S)-oxetan-2-yl;

[0026] R2is selected from hydrogen or halogen;

[0027] R3is selected from hydrogen, C1-C3alkyl or halogen;

[0028] R4is selected from hydrogen, optionally substituted C1-C6alkyl or optionally substituted C3-C7cycloalkyl; when R4is substituted C1-C6alkyl or substituted C3-C7cycloalkyl, the substituents can be on any available point of attachment, and are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, amino, haloalkyl, hydroxyalkyl, carboxyl or carboxylate;

[0029] R5is selected from hydrogen or fluorine;

[0030] X, Z and Z1are each independently selected from -CH or nitrogen.

[0031] In preferred embodiments of the compound of formula (I) above, R1is (S)-oxetan-2-yl; R2is selected from hydrogen or fluorine; R3is selected from hydrogen, methyl, fluorine or chlorine; R4is selected from hydrogen, hydroxymethyl or cyclopropyl; R5is selected from hydrogen or fluorine; X and Z are each independently selected from -CH or nitrogen; and Z1is -CH.

[0032] As further preferred embodiments, the compound of formula (I) above, or a pharmaceutically acceptable salt thereof, is selected from:

[0033]

[0034]

[0035]

[0036] In a second aspect, the present application provides a pharmaceutical composition comprising a compound of the first aspect above, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent and / or excipient.

[0037] In a third aspect, the present application provides the use of a compound of the first aspect above, or a pharmaceutically acceptable salt thereof, in the manufacture of a glucagon-like peptide-1 receptor agonist.

[0038] In a fourth aspect, the present application provides use of a compound of the first aspect or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of the second aspect in the manufacture of a medicament for the prevention and / or treatment of a disease, condition or disorder associated with glucagon-like peptide-1 activity, wherein the disease, condition or disorder is modulated or treated by agonizing glucagon-like peptide-1 receptor.

[0039] In preferred embodiments, the disease, condition or disorder is selected from the group consisting of:

[0040] Type 1 diabetes, Type 2 diabetes, Prediabetes, idiopathic type 1 diabetes, Latent Autoimmune Diabetes in the Adult (LADA), Early Onset Type 2 Diabetes (EOD), Youth-Onset Aypical Diabetes (YOAD), Adult-Onset Diabetes in Youth (MODY), malnutrition-related diabetes, gestational diabetes, hyperglycemia, insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, kidney disease, diabetic retinopathy, adipocyte dysfunction, visceral adipocyte accumulation, sleep apnea, obesity, eating disorders, weight gain due to use of other agents, excessive sugar craving, dyslipidemia, hyperinsulinemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), fibrosis, cirrhosis, hepatocellular carcinoma, cardiovascular disease, atherosclerosis, coronary artery disease, peripheral vascular disease, hypertension, endothelial dysfunction, impaired vascular compliance, congestive heart failure, myocardial infarction, stroke, hemorrhagic stroke, ischemic stroke, traumatic brain injury, pulmonary hypertension, angioplasty restenosis, intermittent claudication, postprandial lipemia, metabolic acidosis, ketosis, arthritis, osteoporosis, Parkinson's disease, left ventricular hypertrophy, peripheral arterial disease, macular degeneration, cataracts, glomerulosclerosis, chronic renal failure, metabolic syndrome, syndrome X, premenstrual syndrome, angina pectoris, thrombosis, atherosclerosis, transient ischemic attack, vascular restenosis, impaired glucose metabolism, impaired fasting glycemic conditions, hyperuricemia, gout, erectile dysfunction, skin and connective tissue abnormalities, psoriasis, foot ulcer, ulcerative colitis, high apo B lipoproteinemia, Alzheimer's disease, schizophrenia, impaired cognitive function, inflammatory bowel disease, short bowel syndrome, Crohn's disease, colitis, irritable bowel syndrome, and polycystic ovary syndrome.

[0041] Further preferably, the disease is Type 2 diabetes or a condition associated therewith selected from the group consisting of hyperglycemia, insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, obesity, dyslipidemia, hypertension, hyperinsulinemia, and nonalcoholic fatty liver disease (NAFLD).

[0042] In a fifth aspect, the present application also provides a method of agonizing a glucagon-like peptide-1 receptor, comprising the step of: administering to a subject in need of agonizing a glucagon-like peptide-1 receptor an effective amount of a compound of the first aspect above or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of the second aspect above.

[0043] In a sixth aspect, the present application also provides a method of preventing and / or treating a disease, condition or disorder associated with glucagon-like peptide-1 activity, comprising: administering to a subject in need thereof a prophylactically and / or therapeutically effective amount of a compound of the first aspect above or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of the second aspect above.

[0044] In particular, the disease, condition or disorder can be modulated or treated by agonizing a glucagon-like peptide-1 receptor.

[0045] In preferred embodiments, the disease, condition or disorder is selected from the group consisting of:

[0046] Type 1 diabetes, Type 2 diabetes, Prediabetes, Idiopathic Type 1 diabetes, Latent Autoimmune Diabetes in Adults (LADA), Early Onset Type 2 Diabetes (EOD), Youth-Onset Aitypical Diabetes (YOAD), Adult-Onset Diabetes in Youth (MODY), Malnutrition-related diabetes, Gestational diabetes, Hyperglycemia, Insulin resistance, Impaired glucose tolerance, Diabetic neuropathy, Diabetic nephropathy, Kidney disease, Diabetic retinopathy, Adipocyte dysfunction, Visceral adipocyte accumulation, Sleep apnea, Obesity, Eating disorders, Weight gain due to use of other agents, Hyperphagia, Dyslipidemia, Hyperinsulinemia, Nonalcoholic fatty liver disease (NAFLD), Nonalcoholic steatohepatitis (NASH), Fibrosis, Cirrhosis, Hepatocellular carcinoma, Cardiovascular disease, Atherosclerosis, Coronary artery disease, Peripheral vascular disease, Hypertension, Endothelial dysfunction, Impaired vascular compliance, Congestive heart failure, Myocardial infarction, Stroke, Hemorrhagic stroke, Ischemic stroke, Traumatic brain injury, Pulmonary hypertension, Restenosis after angioplasty, Intermittent claudication, Postprandial lipemia, Metabolic acidosis, Ketosis, Arthritis, Osteoporosis, Parkinson's disease, Left ventricular hypertrophy, Peripheral arterial disease, Macular degeneration, Cataracts, Glomerulosclerosis, Chronic renal failure, Metabolic syndrome, Syndrome X, Premenstrual syndrome, Angina pectoris, Thrombosis, Atherosclerosis, Transient ischemic attack, Vascular restenosis, Impaired glucose metabolism, Impaired fasting glucose conditions, Hyperuricemia, Gout, Erectile dysfunction, Skin and connective tissue abnormalities, Psoriasis, Foot ulcerations, Ulcerative colitis, High apo B lipoproteinemia, Alzheimer's disease, Schizophrenia, Impaired cognitive function, Inflammatory bowel disease, Short bowel syndrome, Crohn's disease, Colitis, Irritable bowel syndrome, and Polycystic ovary syndrome.

[0047] Further preferably, the disease is Type 2 diabetes or the following conditions associated therewith: hyperglycemia, insulin resistance, impaired glucose tolerance, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, obesity, dyslipidemia, hypertension, hyperinsulinemia, and nonalcoholic fatty liver disease (NAFLD).

[0048] Beneficial effects

[0049] The compound of formula (I) or a pharmaceutically acceptable salt thereof provided by the present application has excellent GLP-1R receptor agonistic activity and obvious blood glucose lowering effect, and has better pharmacokinetic characteristics, thereby providing more choices for the prevention and / or treatment of diseases, conditions or disorders associated with GLP-1 activity, and having better application prospects in clinic. BRIEF DESCRIPTION OF DRAWINGS

[0050] One or more embodiments are illustrated by way of example in the figures that form a part of this disclosure and which do not limit the scope of embodiments in any way. The drawings are illustrative and for use in understanding the principles of embodiments. As such, the drawings should not be considered restrictive of the scope of embodiments. The word "exemplary" is used herein to mean "serving as an example, instance, or illustration." Any implementation described herein as "exemplary" is not necessarily to be construed as preferred or advantageous over other implementations.

[0051] Figure 1 Figure 2 shows the results of a mouse glucose tolerance test (IPGTT test) of compound 2 described in Example 24 of the present application; wherein, panel A shows the effect of compound 2 of the present application on blood glucose level in hGLP-1R mouse glucose tolerance test compared with known control drug PF-06882961, wherein the abscissa shows the treatment time (unit: min), the ordinate shows the blood glucose level (unit: mmol / L), "#" indicates the significance of "hGLP-1R+PF-06882961" group compared with "hGLP-1R+vehicle" group, wherein "##" represents P<0.01, "####" represents P<0.0001; "^" indicates the significance of "hGLP-1R+compound 2" group compared with "hGLP-1R+vehicle" group, wherein "^^" represents P<0.01, "^^^^" represents P<0.0001; panel B shows the AUC of blood glucose level change after glucose administration in glucose tolerance test of compound 2 of the present application compared with known control drug PF-06882961, wherein the abscissa shows the group, the ordinate shows the blood glucose AUC 0-120 (unit: (mmol / L)*min), "####" indicates the significance of "hGLP-1R+PF-06882961" group and "hGLP-1R+compound 2" group compared with "hGLP-1R+vehicle" group, P<0.0001. DETAILED DESCRIPTION

[0052] In order to make the objects, technical solutions, and advantages of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below. Obviously, the described embodiments are some but not all of the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the scope of the present application.

[0053] In addition, in order to better illustrate the present application, numerous specific details are given in the specific embodiments below. Those skilled in the art should understand that the present application can also be implemented without some specific details. In some embodiments, the raw materials, methods, and the like that are well known to those skilled in the art are not described in detail in order to highlight the main idea of the present application.

[0054] Unless explicitly stated otherwise, throughout the specification and claims, the term "comprise" or variations such as "include" or "comprising", etc., will be understood to include stated components but not to exclude other components.

[0055] Definition of terms

[0056] It should be noted that, unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0057] The term "C1-C6 alkyl" refers to an alkyl group containing 1 to 6 carbon atoms, preferably an alkyl group containing 1 to 4 carbon atoms, such as methyl. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be at any available point of attachment and is preferably independently selected from one or more of the following groups: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, amino, haloalkyl, hydroxyalkyl, carboxyl or carboxylate.

[0058] The term "C3-C7 cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 7 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, etc., preferably cyclopropyl, cyclopentyl. Polycyclic cycloalkyls include cycloalkyls of spirocycles, condensed rings, and bridged rings. Cycloalkyls may be optionally substituted or unsubstituted. When substituted, substituents are preferably independently selected from one or more of the following groups: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, amino, haloalkyl, hydroxyalkyl, carboxyl, or carboxylate.

[0059] The term "halogen" refers to fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine.

[0060] The term "optionally" or "optional" means that the subsequently described event or circumstance can or can not occur, and this description includes instances where the event or circumstance occurs and instances where it does not. For example, "heterocycloalkyl optionally substituted with alkyl" means that alkyl can or can not be present, and this description includes instances where the heterocycloalkyl group is substituted with alkyl and instances where the heterocycloalkyl group is not substituted with alkyl.

[0061] "Substituted" means that one or more hydrogen atoms, preferably up to five, more preferably one to three, of a group are independently of each other replaced with a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, which can or cannot be possible, as determined (experimentally or theoretically) by one skilled in the art without undue effort. For example, an amino or hydroxyl group with a free hydrogen can not be stable when bound to a carbon atom with an unsaturated (e.g., olefinic) bond.

[0062] Pharmaceutically acceptable salts of the compounds of formula (I) and processes for their preparation

[0063] It is to be understood that the pharmaceutically acceptable salts of the compounds of formula (I) according to the present application include both acid addition salts and base salts.

[0064] Suitable acid addition salts are formed from acids which form nontoxic salts. Examples of acid addition salts include acetate, adipate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate, borate, camsylate, citrate, cyclohexylsulfamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / diphosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, 1,5- naphthalenedisulfonate, and xinafoate salts.

[0065] Suitable base salts are formed from bases which form nontoxic salts. Examples of base salts include aluminum, arginine, benzathine, calcium, choline, diethylamine, bis(2-hydroxyethyl)amine (diolamine), glycine, lysine, magnesium, meglumine, 2-aminoethanol (olamine), potassium, sodium, 2-amino-2-(hydroxymethyl)-propane-1,3-diol (tris or tromethamine), and zinc salts.

[0066] The compounds of formula (I) of the present application can also form acid and base "hemisalts", e.g. hemisulphate and hemicalcium salts. For a review on suitable salts see Handbook of Pharmaceutical Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley-VCH, 2002).

[0067] The pharmaceutically acceptable salts of the compounds of formula (I) of the present application can be prepared by one or more of three methods:

[0068] (i) by reacting a compound of formula (I) with the desired acid or base;

[0069] (ii) by removing an acid- or base-labile protecting group from a suitable precursor of formula (I) using the desired acid or base, or by ring opening an appropriate cyclic precursor (e.g. a lactone or lactam); or

[0070] (iii) by converting one salt of formula (I) to another using the desired acid or base, or by means of an appropriate ion exchange column.

[0071] Preparation of compounds of formula (I)

[0072] The compounds of formula (I) of the present application can be prepared using well known common general knowledge of the synthetic organic chemist using the general and specific methods described herein. Such common general knowledge can be found in standard reference works, such as Comprehensive Organic Chemistry, Ed. Barton and Ollis, Elsevier; Comprehensive Organic Transformations: A Guide to Functional Group Preparations, Larock, John Wiley and Sons; and Compendium of Organic Synthetic Methods, Vol. I-XII (published by Wiley-Interscience). The starting materials used in the present application are either commercially available or can be prepared by routine methods known in the art.

[0073] Some specific preparation examples of the compounds of formula (I) of the present application are provided below, together with performance, efficacy experiments.

[0074] Example 1, synthesis of compound 1

[0075]

[0076] The specific steps are as follows:

[0077] Step 1-1 : Synthesis of compound 1C

[0078] Compound 1A (5 g, 18.9 mmol), compound 1B (7.0 g, 22.6 mmol), Pd(dppf)Cl2(0.5 g, 0.68 mmol), potassium carbonate (5.2 g, 37.6 mmol) were added into a 100 mL microwave tube in turn, and then mixed solvents 1,4-dioxane (50 mL) and water (5 mL) were added. Microwave heating was carried out at 120°C for 3 hours under argon atmosphere. The reaction solution was concentrated and directly separated by silica gel column, eluted with 0-20% EA / PE, and the corresponding solution was collected and concentrated to obtain a colorless oil, which was compound 1C (6.0 g). LCMS (ESI+): 367 (M+H) + .

[0079] Step 1-2: Synthesis of compound 1D

[0080] Compound 1C (6 g, 16.3 mmol) obtained in step 1-1 was dissolved in methanol (60 mL) in a 150 mL round-bottom flask, Pd / C (10%, 50% water) (0.6 g) was added, hydrogen was replaced for 3 times, and stirring was continued for 16 hours. TLC detection showed that the reaction was complete. Filtration, concentration of the mother liquor, and beating with a mixture of petroleum ether and ethyl acetate (50 mL, 5:1, v:v) for 2 hours, and then filtration of the white solid precipitated, which was compound 1D (2.0 g). LCMS (ESI+): 279 (M+H) + .

[0081] Step 1-3: Synthesis of compound 1F

[0082] Compound 1E (1.42 g, 7 mmol, purchased from Bide Pharm, cat.#: BD10249), Pd(PPh3)2Cl2(98 mg, 0.14 mmol), PPh3(28 mg, 0.105 mmol) were added into a 50 mL round-bottom flask in turn, and then THF (22 mL) was added. After nitrogen replacement for 3 times, stirring was carried out at room temperature (25°C) for 20 minutes, and then triethylamine (1.89 mL, 13.65 mmol) and trimethylsilyacetylene (1.09 mL, 7.7 mmol) were added. After stirring for 20 minutes, cuprous iodide (40 mg, 0.21 mmol) was added, and stirring was carried out at room temperature for 5 hours. TLC detection showed that the reaction was complete. The reaction solution was concentrated to obtain a brown solid residue. After addition of n-pentane (50 mL) and stirring for 10 minutes, filtration was carried out, and the mother liquor was concentrated to obtain a yellow solid, which was compound 1F (1.45 g) and was directly used in the next step.

[0083] Step 1-4: Synthesis of compound 1G

[0084] Compound 1F (1.45 g, 6.6 mmol) obtained from step 1-3 was added to a 50 mL round bottom flask, ethanol (25 mL) was added to get a light yellow solution, cooled to 0-5 °C with stirring, sodium borohydride (0.25 g, 6.6 mmol) was added portion wise, gas was evolved, stirring was continued for 30 minutes, TLC showed the reaction was complete. Saturated ammonium chloride solution (50 mL) was added drop wise, after 10 minutes stirring, dichloromethane was added to extract the aqueous layer, the aqueous layer was extracted twice more with dichloromethane. The organic layers were combined, dried over magnesium sulfate, filtered, concentrated, purified on silica gel column (0-50% DCM / PE) elution, the corresponding fractions were collected and concentrated to get a colorless oil, which was compound 1G (1.3 g).

[0085] Step 1-5: Synthesis of compound 1H

[0086] Compound 1D (0.28 g, 1 mmol), compound 1G (0.22 g, 1 mmol), PPh3 (0.39 g, 1.5 mmol) obtained from the above steps were added to a 50 mL round bottom flask, anhydrous tetrahydrofuran (15 mL) was added to get a mixture solution, cooled to 0-5 °C with stirring after 3 times nitrogen replacement, DEAD (0.26 g, 1.5 mmol) was added drop wise carefully, temperature was controlled below 5 °C, after addition, stirring was continued for 16 hours, TLC showed the reaction was complete. The reaction mixture was concentrated directly on silica gel column, elution with 0-20% EA / PE, the corresponding fractions were collected and concentrated to get a colorless oil, which was compound 1H (0.20 g). LCMS (ESI+): 427 (M+H) + .

[0087] Step 1-6: Synthesis of compound 11

[0088] Compound 1H (0.20 g, 0.41 mmol) obtained from step 1-5 was dissolved in dichloromethane (10 mL) in a 25 mL round bottom flask, trifluoroacetic acid (2 mL) was added with stirring, gas was evolved, after stirring for 16 hours, it was concentrated directly, sodium bicarbonate solution was added, pH was adjusted to 8-9, dichloromethane was extracted 3 times, the organic layers were combined, dried over anhydrous magnesium sulfate, filtered and concentrated to get an oil, which was compound 11 (0.11 g), which was used directly in the next step. LCMS (ESI+): 298 (M+H) + .

[0089] Step 1-7: Synthesis of compound 1K

[0090] Compound 1I (0.10 g, 0.34 mmol, prepared according to step 1-6) was dissolved in acetonitrile (10 mL) in a 25 mL round bottom flask, potassium carbonate (0.09 g, 0.67 mmol) was added with stirring, after 10 minutes compound 1J (0.09 g, 0.34 mmol) was added and heated to 45-50 °C, the reaction was complete after 3 hours as monitored by TLC. The reaction mixture was directly concentrated to dryness and column chromatography was performed with 0-5% MeOH / DCM as eluent, the fractions were collected and concentrated to get compound 1K (0.10 g) as colorless oil. LCMS (ESI+): 569 (M+H) + .

[0091] Step 1-8: Synthesis of compound 1

[0092] Compound 1K (0.10 g, 0.17 mmol) obtained from step 1-7 was dissolved in tetrahydrofuran (5 mL) and water (1.5 mL) in a 25 mL round bottom flask, lithium hydroxide (5 mg, 0.21 mmol) was added with stirring. The reaction was carried out at room temperature for 16 hours. Acetic acid was added to adjust the pH to 5-6, stirred for 5 minutes, extracted with dichloromethane three times, the organic phases were combined, dried over anhydrous magnesium sulfate, filtered, and concentrated to get compound 1 (30 mg) as colorless oil. LCMS (ESI+): 555 (M+H) + .

[0093] Example 2, synthesis of compound 2

[0094]

[0095] The detailed procedure is as follows:

[0096] Step 2-1 : Synthesis of compound 2A

[0097] Compound 1G (0.3 g, 1.35 mmol, prepared according to step 1-4 in example 1), 2-chloro-6-hydroxypyridine (0.19 g, 1.47 mmol), PPh3 (0.5 g, 1.90 mmol) were added to a 25 mL round bottom flask successively to get a solution in anhydrous tetrahydrofuran (10 mL). Nitrogen was purged for 3 times, stirred and cooled to 0-5 °C, DEAD (0.35 g, 1.90 mmol) was added dropwise carefully, the temperature was controlled below 5 °C, after the addition was completed, the stirring was continued for 16 hours, the reaction was complete as monitored by TLC. The reaction mixture was concentrated to dryness and directly column chromatography was performed with 0-20% EA / PE as eluent, the fractions were collected and concentrated to get compound 2A (0.25 g) as colorless oil. LCMS (ESI+): 334 (M+H) + .

[0098] Step 2-2: Synthesis of compound 2B

[0099] Compound 2A (0.20 g, 1.35 mmol) prepared according to step 2-1, N-Boc piperazine (0.13 g, 1.47 mmol), Pd2(dba)3(30 mg), Xantphos (37 mg, 1.90 mmol), potassium carbonate (0.13 g) were added into a 10 mL microwave tube in turn, 1,4-dioxane (5 mL) was added, and the tube was capped after bubbling with argon for 30 seconds. The mixture was heated at 110 °C for 2 hours. The reaction was complete according to TLC detection. The reaction solution was concentrated to dryness and directly purified by silica gel column separation. The corresponding solution was collected and concentrated to dryness to obtain colorless oil, which was compound 2B (0.16 g). LCMS (ESI+): 484 (M+H) + .

[0100] Step 2-3: Synthesis of compound 2C

[0101] Compound 2B (160 mg, 0.33 mmol) prepared according to step 2-2 was dissolved in dichloromethane (5 mL) in a 25 mL round-bottom flask. Trifluoroacetic acid (2 mL) was added with stirring, and gas bubbles were generated. The reaction was continued to stir at room temperature for 6 hours. The reaction was complete according to TLC detection. Compound 2C (80 mg) was obtained after concentration. It was directly used in the next step. LCMS (ESI+): 312 (M+H) + .

[0102] Step 2-4: Synthesis of compound 2D

[0103] Compound 2C (80 mg, 0.26 mmol) prepared according to step 2-3 was dissolved in acetonitrile (5 mL) in a 10 mL round-bottom flask. Potassium carbonate (100 mg, 0.72 mmol) was added with stirring, and the reaction was continued to stir at room temperature for 10 minutes. Compound 1J (80 mg, 0.27 mmol) was added, and the reaction was heated to 50-55 °C for 2 hours. The reaction was complete according to TLC detection. After concentration to dryness, it was directly purified by preparative plate. The corresponding fluorescent silica gel was collected and eluted, and colorless oil was obtained after concentration, which was compound 2D (40 mg). LCMS (ESI+): 570 (M+H) + .

[0104] Step 2-5: Synthesis of compound 2

[0105] Compound 2D (40 mg, 0.07 mmol) prepared in step 2-4 was dissolved in tetrahydrofuran (5 mL) and water (1 mL) in a 10 mL round bottom flask. Lithium hydroxide (2.4 mg, 0.1 mmol) was added with stirring and the reaction was allowed to proceed at room temperature for 16 hours. The reaction was monitored by TLC. Three drops of acetic acid were added and stirred for 5 minutes. After drying by concentration, the product was purified by direct prep plate elution with 10% MeOH / DCM. The fractions corresponding to the fluorescent silica gel were collected and concentrated to give white solid, which was compound 2 (35 mg). LCMS (ESI+): 556 (M+H) + .

[0106] Example 3, Synthesis of Compound 3

[0107]

[0108] Following the synthetic route of Example 1, in step 1-3, propynyl was used in place of trimethylsilylethynyl to give compound (4-propynyl-2-fluorophenyl)methanol; and following the synthetic route of Example 2, in step 2-1, compound (4-propynyl-2-fluorophenyl)methanol was used in place of compound 1G to give compound 3. LCMS (ESI+): 570.43 (M+H)+.

[0109] Example 4, Synthesis of Compound 4

[0110]

[0111] Following the synthetic route of Example 1, in step 1-3, cyclopropylethynyl was used in place of trimethylsilylethynyl to give compound (4-cyclopropylethynyl-2-fluorophenyl)methanol; and following the synthetic route of Example 2, in step 2-1, compound (4-cyclopropylethynyl-2-fluorophenyl)methanol was used in place of compound 1G to give compound 4. LCMS (ESI+): 596.35 (M+H)+.

[0112] Example 5, Synthesis of Compound 5

[0113]

[0114] The specific steps are as follows:

[0115] Step 5-1 : Synthesis of compound 5E

[0116] Following the synthetic route of Example 1, in step 1-3, 2-(2-propargyloxy)tetrahydropyran was used in place of trimethylsilylethynyl to give compound 5C; and following the synthetic route of Example 2, in step 2-1, compound 5C was used in place of compound 1G to give compound 5E. LCMS (ESI+): 526.47 (M+H)+.

[0117] Step 5-2: Synthesis of compound 5F

[0118] The compound 5E (120 mg, 0.23 mmol) obtained from step 5-1 was added into a mixed solvent of dichloromethane (6 mL) and methanol (0.5 mL), and trifluoroacetic acid (2 mL) was added. The reaction solution was stirred at room temperature for about 2 hours. The reaction solution was concentrated under reduced pressure to obtain compound 5F in the form of light yellow oil. The crude product was not further purified and was directly used in the next step reaction. LCMS (ESI+): 342.06 (M+H)+.

[0119] Step 5-3: Synthesis of compound 5

[0120] According to the synthetic route of Example 2, in step 2-4, compound 5F was used instead of compound 2C to obtain compound 5. LCMS (ESI+): 586.37 (M+H)+.

[0121] Example 6, synthesis of compound 6

[0122]

[0123] The specific steps are as follows:

[0124] Step 6-1 : Synthesis of compound 6B

[0125] Compound 6A (10.8 g, 0.05 mol) was added into tetrahydrofuran (100 mL), and after stirring and dissolving, N,N-diisopropyl ethylamine (32.3 g, 0.25 mol) was added, and then cooled to 0°C. (S)-oxetan-2-methylamine (4.7 g, 0.053 mol) was added dropwise, and after the addition was completed, the temperature was slowly increased to 25°C, and stirred for 1 hour, then 100 mL of ethyl acetate was added, and the organic phase was washed with saturated brine three times, then dried over anhydrous sodium sulfate, filtered, and the mother liquor was concentrated to obtain a yellow solid, which was compound 6B (13.5 g), which was directly used in the next step.

[0126] Step 6-2: Synthesis of compound 6C

[0127] Compound 6B (13.5 g, 0.047 mol) was added into methanol (150 mL), and after stirring and dissolving, 10% Pd / C (3.5 g, 50% water) was added, and after stirring under hydrogen gas for 3 times, the reaction was stirred at room temperature for 18 hours, then filtered, the filter cake was washed with methanol twice, and the organic phase was combined and concentrated to obtain a dark oil, which was compound 6C (11 g), which was directly used in the next step.

[0128] Step 6-3: Synthesis of compound 6D

[0129] Compound 6C (11 g, 0.043 mol) was added to THF (100 mL) and stirred to dissolve, then 2-chloro-l, l, l-trimethoxyethane (10 g, 0.064 mol) and p-toluenesulfonic acid (0.83 g, 0.005 mol) were added. After addition, the mixture was heated to 60 °C for 2 hours. After cooling to room temperature, the mixture was directly concentrated and purified by column chromatography with 0-5% MeOH / DCM to give compound 6D (8 g) as an oil, which was used directly in the next step. LCMS (ESI+): 313.1 (M+H) + .

[0130] Step 6-4: Synthesis of compound 6

[0131] Following the synthetic route of Example 2, in step 2-4, compound 6D was used in place of compound 1J to give compound 6. LCMS (ESI+): 573.4 (M+H)+.

[0132] Example 7, Synthesis of Compound 7

[0133]

[0134] Following the synthetic route of Example 2, in step 2-1, compound 4-(trimethylsilyl ethynyl)benzyl methanol was used in place of compound 1G to give compound 7. LCMS (ESI+): 536.4 (M+H)+.

[0135] Example 8, Synthesis of Compound 8

[0136]

[0137] The specific steps are as follows:

[0138] Step 8-1 : Synthesis of compound 8D

[0139] Following the synthetic route of Example 1, in step 1-3, compound 8A was used in place of compound 1E to give compound 8C; then following the synthetic route of Example 2, in step 2-1, compound 8C was used in place of compound 1G, and 2-bromo-6-hydroxypyridine was used in place of 2-chloro-6-hydroxypyridine to give compound 8D. LCMS (ESI+): 394.15 (M+H)+.

[0140] Step 8-2: Synthesis of compound 8E

[0141] Compound 8D (500 mg, 1.27 mmol) obtained from step 8-1, N-Boc piperazine (283 mg, 1.52 mmol), Pd2(dba)3(58 mg, 0.064 mmol), RuPhos (59 mg, 0.13 mmol) and cesium carbonate (620 mg, 1.91 mmol) were added into tetrahydrofuran (10 mL) successively. After three times of argon replacement, the reaction was heated to 70 °C for about 4 hours. After the reaction was cooled to room temperature, it was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluted with 0-15% ethyl acetate / petroleum ether, and the corresponding eluent was collected and concentrated to obtain compound 8E (610 mg) as a colorless oil. LCMS (ESI+): 500.10 (M+H)+.

[0142] Step 8-3: Synthesis of compound 8

[0143] Following the synthetic route of Example 2, in step 2-3, compound 8E was used to replace compound 2B to obtain compound 8. LCMS (ESI+): 572.10 (M+H)+.

[0144] Example 9, synthesis of compound 9

[0145]

[0146] The specific steps are as follows:

[0147] Step 9-1 : Synthesis of compound 9B

[0148] Compound 9A (5.00 g, 23.1 mmol), (S)-oxetan-2-ylmethylamine (2.41 g, 27.7 mmol) and triethylamine (4.67 g, 46.2 mmol) were added into acetonitrile (200 mL) successively. The reaction was stirred at 50 °C for about 4 hours. After the reaction was cooled to room temperature, it was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluted with 0-20% ethyl acetate / petroleum ether, and the corresponding solution was collected and concentrated to obtain compound 9B (5.2 g) as a yellow solid. LCMS (ESI+): 268.21 (M+H)+.

[0149] Step 9-2: Synthesis of compound 9C

[0150] Compound 9B (4.5 g, 18.0 mmol) obtained from step 9-1 was added to a 100 mL round bottom flask, dissolved in methanol (50 mL), and Pd / C (2.0 g, 10 w%, 50% water) was added. The reaction flask was replaced with hydrogen three times, and the reaction was allowed to proceed under a hydrogen atmosphere for about 3 hours. The reaction was filtered through celite, and the filtrate was concentrated under reduced pressure to obtain a yellow solid, which was compound 9C (3.7 g). LCMS (ESI+): 238.05 (M+H)+.

[0151] Step 9-3: Synthesis of compound 9D

[0152] Compound 9C (3.7 g, 15.6 mmol) obtained from step 9-2 was dissolved in tetrahydrofuran (50 mL), and a solution of chloroacetic anhydride (2.93 g, 17.2 mmol) in tetrahydrofuran (10 mL) was added with rapid stirring at room temperature. The reaction was allowed to warm to 55 °C and was allowed to proceed for about 6 hours. The reaction was cooled to room temperature and was poured into a mixture of ethyl acetate (150 mL) and saturated aqueous sodium bicarbonate (200 mL). The organic phase was collected. The organic phase was washed with saturated aqueous sodium chloride (100 mL) and was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel, eluting with 0-80% ethyl acetate / petroleum ether, and the corresponding fractions were collected and concentrated to obtain a light yellow solid, which was compound 9D (4.1 g), which was used directly in the next step. LCMS (ESI+): 296.30 (M+H)+.

[0153] Step 9-4: Synthesis of compound 9

[0154] Following the synthetic route of Example 2, in step 2-4, compound 9D was used in place of compound 1J to obtain compound 9. LCMS (ESI+): 557.10 (M+H)+.

[0155] Example 10, Synthesis of Compound 10

[0156]

[0157] Following the synthetic route of Example 2, in step 2-1, 6-chloro-3-fluoropyridin-2-ol was used in place of 2-chloro-6-hydroxypyridine to obtain compound 10. LCMS (ESI+): 574.2 (M+H)+.

[0158] Example 11, Synthesis of Compound 11

[0159]

[0160] Following the synthetic route of Example 1, in Step 1-7, compound 6D was used instead of compound 1J to obtain compound 11. LCMS (ESI+): 573.3 (M+H)+.

[0161] Example 12, Synthesis of compound 12

[0162]

[0163] Following the synthetic route of Example 2, in Step 2-3, compound 8E was used instead of compound 2B to obtain compound 8F; in Step 2-4, compound 8F was used instead of compound 2C, and compound 6D was used instead of compound 1J to obtain compound 12. LCMS (ESI+): 590.11 (M+H)+.

[0164] Example 13, Synthesis of compound 13

[0165]

[0166] Following the synthetic route of Example 2, in Step 2-3, compound 8E was used instead of compound 2B to obtain compound 8F; in Step 2-4, compound 8F was used instead of compound 2C, and compound 9D was used instead of compound 1J to obtain compound 13. LCMS (ESI+): 573.21 (M+H)+.

[0167] Example 14, Synthesis of compound 14

[0168]

[0169] The specific steps are as follows:

[0170] Following the synthetic route of Example 1, in Step 1-3, compound 14A was used instead of compound 1E to obtain compound 14C;

[0171] Following the synthetic route of Example 2, in Step 2-1, compound 14C was used instead of compound 1G, and 6-chloro-3-fluoropyridin-2-ol was used instead of 2-chloro-6-hydroxypyridine; in Step 2-4, compound 9D was used instead of compound 1J to obtain compound 14. LCMS (ESI+): 571.3 (M+H)+.

[0172] Example 15, Synthesis of compound 15

[0173]

[0174] Following the synthetic route of Example 2, in step 2-1, compound 1G is replaced with 4-(trimethylsilylethynyl)benzyl methanol, 2-chloro-6-hydroxypyridine is replaced with 6-chloro-3-fluoropyridin-2-ol; in step 2-4, compound 1J is replaced with compound 9D, to give compound 16. LCMS (ESI+): 557.31 (M+H)+.

[0175] Example 16, Synthesis of compound 16

[0176]

[0177] Following the synthetic route of Example 2, in step 2-1, compound 1G is replaced with 4-(trimethylsilylethynyl)benzyl methanol, 2-chloro-6-hydroxypyridine is replaced with 6-chloro-3-fluoropyridin-2-ol; in step 2-4, compound 1J is replaced with compound 9D, to give compound 16. LCMS (ESI+): 557.31 (M+H)+.

[0178] Example 17, Synthesis of compound 17

[0179]

[0180] Following the synthetic route of Example 2, in step 2-1, 2-chloro-6-hydroxypyridine is replaced with 6-chloro-3-fluoropyridin-2-ol; in step 2-4, compound 1J is replaced with compound 9D, to give compound 17. LCMS (ESI+): 575.40 (M+H)+.

[0181] Example 18, Synthesis of compound 18

[0182]

[0183] Following the synthetic route of Example 1, in step 1-3, compound 1E is replaced with compound 8A, to give compound 8C. Then following the synthetic route of Example 2, in step 2-1, compound 1G is replaced with compound 8C, 2-chloro-6-hydroxypyridine is replaced with 6-chloro-3-fluoropyridin-2-ol; in step 2-4, compound 1J is replaced with compound 9D, to give compound 18. LCMS (ESI+): 591.2 (M+H)+.

[0184] Example 19, Synthesis of compound 19

[0185]

[0186] Following the synthetic route of Example 1, in Step 1-3, compound 8A was used instead of compound 1E to give compound 8C. Following the synthetic route of Example 2, in Step 2-1, compound 8C was used instead of compound 1G, and 6-chloro-3-fluoropyridin-2-ol was used instead of 2-chloro-6-hydroxypyridine to give compound 19. LCMS (ESI+): 590.2 (M+H)+.

[0187] Example 20, Synthesis of compound 20

[0188]

[0189] The specific steps are as follows:

[0190] Step 20-1 : Synthesis of compound 20B

[0191] Compound 20A (2.00 g, 13.6 mmol), compound IB (8.38 g, 27.1 mmol), Pd(dtbpf)Cl2 (90 mg, 0.14 mmol) and potassium carbonate (7.49 g, 54.2 mmol) were added into a mixture solvent of dioxane (30 mL) and water (5 mL) successively. The reaction solution was replaced with argon for three times, and then the reaction solution was heated to 100 °C for about 3 hours. After the reaction solution was cooled to room temperature, it was poured into ethyl acetate (150 mL), and the pH value of the mixture was adjusted to about 7 with acetic acid. The organic phase was separated, and dried with anhydrous sodium sulfate; after the solid was removed by filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluted with 0-60% ethyl acetate / petroleum ether, and the corresponding eluate was collected and concentrated. Compound 20B (1.66 g) was obtained as a light brown solid. LCMS (ESI+): 295.21 (M+H)+.

[0192] Step 20-2: Synthesis of compound 20C

[0193] Compound 20B (1.10 g, 3.74 mmol) obtained from Step 20-1 was added into a 100 mL round-bottom flask, dissolved with methanol (25 mL), and then Pd / C (500 mg, 10 w%, containing 50% water) was added. The reaction flask was replaced with hydrogen for three times, and the reaction solution was reacted under hydrogen atmosphere for about 16 hours. The reaction solution was filtered through diatomite, and the filtrate was concentrated under reduced pressure to obtain white solid, which was compound 20C (1.10 g) and was used directly in the next step. LCMS (ESI+): 297.31 (M+H)+.

[0194] Step 20-3: Synthesis of compound 20

[0195] Following the synthetic route of Example 1, in Step 1-5, compound 20C was used instead of compound 1D, and in Step 1-7, compound 9D was used instead of compound 1J, to obtain compound 20. LCMS (ESI+): 574.40 (M+H)+.

[0196] Example 21, Synthesis of compound 21

[0197]

[0198] Following the synthetic route of Example 1, in Step 1-5, compound 20C was used instead of compound 1D, to obtain compound 21. LCMS (ESI+): 573.09 (M+H)+.

[0199] Example 22, GLP-1R agonist activity assay

[0200] The GLP-1R agonist activity of compounds 1-21 of the present application and known control drug PF-06882961 was determined using GLP-1R cAMP assay.

[0201] 1. Reagent consumables

[0202] (1) Reagents

[0203]

[0204]

[0205] (2) Cell information

[0206]

[0207] (3) Equipment

[0208]

[0209] 2. Experimental methods

[0210] 2.1 Compound source plate configuration

[0211] Using an automated liquid handling platform, compounds 1-21 of the present application, known control drug PF-06882961 and positive control drug Exendin-4 (to monitor consistency of the assay) were diluted, respectively, with a starting concentration of 500 nM, 4-fold dilution gradient, a total of 10 concentrations, and dimethyl sulfoxide as diluent.

[0212] 2.2 Transferring compounds

[0213] 1) Transfer 100 nL of each compound to OptiPlate 384 well plate using Echo Pipetting system;

[0214] 2) Centrifuge 384 well plate at 1000 rpm for 5 seconds.

[0215] 2.3 Preparing cell suspension

[0216] 1) Resuspend one vial of GLP-1R-HEK293 cell line according to the regular cell resuspension procedure in a 37°C water bath;

[0217] 2) Transfer the cell suspension to a 15 mL centrifuge tube containing 10 mL of 1 x HBSS solution;

[0218] 3) Centrifuge at 1000 rpm for 5 minutes at room temperature to collect the cells;

[0219] 4) Carefully aspirate the supernatant, taking care not to aspirate the cells;

[0220] 5) Gently flick the centrifuge tube to loosen the cells and add 10 mL of 1 x HBSS solution to resuspend the cells. Use a sterile pipette to pipette the cells up and down to break up the cells into a single cell suspension;

[0221] 6) Count the cells using a Vi-Cell counter to determine the cell concentration and to determine the cell viability;

[0222] 7) Resuspend the GLP-1R-HEK293 cells in cAMP assay buffer (from the cAMP assay kit described above) to a concentration of 2.0 x 10 5 / mL;

[0223] 8) Transfer 10 μL of GLP-1R-HEK293 cells to OptiPlate 384 well plate.

[0224] 2.4 HTRF cAMP agonist assay

[0225] 1) Incubate the 384 well plate for 20 minutes at room temperature followed by the addition of cAMP assay reagent (from the cAMP assay kit described above);

[0226] 2) Add 10 μL of cAMP assay reagent per well using an electronic multichannel pipette;

[0227] 3) Cover the 384 well plate with a TopSeal-A membrane and incubate for 60 minutes at room temperature;

[0228] 4) Remove the TopSeal-A membrane and read the signal on an Envision instrument.

[0229] 3. Experimental results

[0230] The GLP-1R cAMP assay results (expressed as EC50) of the compounds 1-21 of the present application and the known control drug PF-06882961 are shown in Table 1 below.

[0231] Table 1

[0232] Compound number EC50 (nM) 1 0.82 2 0.96 3 0.80 4 1.75 5 5.20 6 0.34 7 0.39 8 0.10 9 0.15 10 0.10 11 0.14 12 0.41 13 0.36 14 1.30 15 0.85 16 1.07 17 0.56 18 0.37 19 0.51 20 0.59 21 0.57 PF-06882961 0.35

[0233] As can be seen from Table 1, in the GLP-1R cAMP experiment, all the compounds show effective GLP-1R agonistic effect, and the compounds 1-21 of the present application can be used as effective GLP-1R receptor agonists.

[0234] Example 23, mouse oral pharmacokinetics

[0235] The compounds 1, 2, 6, 7, 8, 9, 10, 11, 13, 17, 18, 19 of the present application and the known control drug PF-06882961 were subjected to pharmacokinetic test using the following experimental procedure:

[0236] 1. Experimental animals

[0237] Healthy male C57 mice (9 for each compound, n=3), aged 6-8 weeks, were purchased from Shanghai Jihui Laboratory Animal Co. LTD.

[0238] 2. Method

[0239] The mice were fasted overnight before oral administration, and free water was provided, and the food was restored 4 hours after administration. According to the weight of the mice before administration, the administration amount was calculated. The gavage administration was performed at a dose of 10 mg / kg. At 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h and 24 h after administration of the test animals, about 110 μL of whole blood was collected from the submandibular vein of the mice by semi-continuous method, and placed in a test tube containing K2EDTA, and centrifuged (2000g, 5 min, 4℃) within 30 min, and the plasma was collected, transferred to a properly labeled test tube, and stored in a -80℃ refrigerator.

[0240] 3. Chromatographic and mass spectrometric conditions

[0241] Chromatographic column: Waters ACQUITY UPLC BEH C18 (2.1 x 50 mm, 1.7 μm); mobile phase A: H2O-0.025% FA-1 mM NH4OAC, mobile phase B: ACN-0.025% FA-1 mM NH4OAC, flow rate: 0.60 mL / min, gradient elution program: initial, 2% B, 0.2 min; 60% B, 0.7 min; 90% B, 1.50 min; 90% B, 2.0 min; 2% B, 2.01 min; 2.6 min, stop; column temperature: 50 °C, injection volume: 1 μL;

[0242] Mass spectrometry method: LC-MS / MS-49 (Triple Quad 6500+) (SCIEX, USA), ion source: ESI source, detection mode: positive ion detection, scanning mode: multiple reaction monitoring (MRM) mode, m / z: 446.20 / 321.10 Da (Glipizide, internal standard).

[0243] 4. Preparation of plasma samples

[0244] Take 10 μL of plasma sample and add internal standard working solution (Glipizide, 50 ng / mL) 200 μL, vortex for 1 min, centrifuge at 5800 rpm for 10 min, transfer 70 μL of supernatant to a 96-well plate, and inject 1 μL of supernatant into LC-MS / MS for analysis.

[0245] 5. Analysis of results

[0246] Phoenix WinNonlin 7.0 was used to calculate the pharmacokinetic (PK) parameters; the mouse oral pharmacokinetic parameters were estimated by a non-compartment model, including AUC, C max , T max , T 1 / 2 , etc. The mouse oral pharmacokinetic parameter results of compounds 1, 2, 6, 7, 8, 9, 10, 11, 13, 17, 18, 19 and PF-06882961 are shown in Table 2 below.

[0247] Table 2

[0248]

[0249] As can be seen from Table 2, compared with the known control drug (PF-06882961), the compounds 1, 2, 6, 7, 8, 9, 10, 11, 13, 17, 18, 19 of the present application have a longer half-life (T 1 / 2 ) and / or have a higher total exposure (AUC).

[0250] Example 24, glucose tolerance test of compound 2 of the present application in mice

[0251] The specific steps are as follows:

[0252] 1) After the experimental animals (hGLP-1R transgenic mice B-hGLP-1R, SPF level mice, male, 6-8 weeks; purchased from Bao Saiguang Biotechnology Co., Ltd. Jiangsu) arrived, all the experimental animals were divided into single-cage feeding and adapted for 7 days (during which the health of the animals was observed) before the experiment started. At least 3 days before the start of the IPGTT experiment, the experimental animals were weighed and given solvent (0.5% methyl cellulose solution) by gavage and intraperitoneal saline injection every day.

[0253] 2) On the day before the experiment, the experimental animals were fasted overnight (14h, 19:00-9:00), and at 9:00am the experimental animals were weighed and blood glucose was measured by tail tip blood sampling. According to the blood glucose and body weight data, the experimental animals were divided into 3 groups:

[0254] the "hGLP-1R+solvent" group in Figure 1 , the "hGLP-1R+PF-06882961" group in Figure 1 , and the "hGLP-1R+compound 2" group in Figure 1 .

[0255] 3) The grouped experimental animals were given 10mL / kg 0.5% methyl cellulose solution, 10mg / kg PF-06882961, and 10mg / kg compound 2 by oral gavage (09:30am), respectively.

[0256] 4) 15min after oral gavage administration (09:45am), the experimental animals were injected intraperitoneally with 2g / kg 40% dextrose solution.

[0257] 5) 1min before glucose solution injection (09:44am), 15min (10:00am), 30min (10:15am), 45min (10:30am), 60min (10:45am), 90min (11:15am), and 120min (11:45am) after glucose solution injection, blood was taken by tail tip blood sampling and blood glucose values were detected, and the results are shown in Figure 1 .

[0258] As can be seen from Figure 1 , compound 2 of the present application has a significant effect of reducing blood glucose, and the overall effect of reducing blood glucose is better than that of PF-06882961.

[0259] It should be pointed out finally that the above embodiments are only used to illustrate the technical solutions of the present application, but not to limit the same; and although the present application has been described in detail with reference to the foregoing embodiments, it should be appreciated by those skilled in the art that the technical solutions recorded in the foregoing embodiments can be modified, or some technical features thereof can be replaced equivalently; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof: in, R1 is (S)-oxetan-2-yl; R2 is selected from hydrogen or halogen; R3 is selected from hydrogen, C1-C6 alkyl or halogen; R4 is selected from hydrogen, C1-C6 alkyl or C3-C7 cycloalkyl; wherein the C1-C6 alkyl is unsubstituted or substituted with hydroxyl; R5 is selected from hydrogen or halogen; X, Z and Z1 are each independently selected from CH or nitrogen.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R1 is (S)-oxetan-2-yl; R2 is selected from hydrogen or halogen; R3 is selected from hydrogen, C1-C3 alkyl or halogen; R4 is selected from hydrogen, C1-C6 alkyl or C3-C7 cycloalkyl; wherein the C1-C6 alkyl is unsubstituted or substituted with hydroxyl; R5 is selected from hydrogen or fluorine; X, Z and Z1 are each independently selected from CH or nitrogen.

3. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein R1 is (S)-oxetan-2-yl; R2 is selected from hydrogen or fluorine; R3 is selected from hydrogen, methyl, fluorine or chlorine; R4 is selected from hydrogen, methyl, hydroxymethyl or cyclopropyl; R5 is selected from hydrogen or fluorine; X and Z are each independently selected from CH or nitrogen; Z1 is CH.

4. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, which is selected from:

5. A pharmaceutical composition comprising the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

6. The pharmaceutical composition according to claim 5, wherein The carrier is an excipient.

7. Use of the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof in the preparation of a glucagon-like peptide-1 receptor agonist.

8. Use of the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to any one of claims 5 to 6, in the preparation of a medicament for preventing and / or treating a disease, condition or disorder associated with glucagon-like peptide-1 activity, wherein: Such diseases, conditions or disorders may be modulated or treated by agonizing the glucagon-like peptide-1 receptor.

9. The use according to claim 8, wherein The disease, condition or disorder is selected from the group consisting of: Type 2 diabetes, insulin resistance, and obesity.

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