Selenium-containing compounds and use thereof

Abstract

Selenium-containing compounds and the use thereof. The compounds have complete GLP-1R-cAMP signaling pathway activation effect (Emax), and the GLP-1R-cAMP median effective concentrations (EC50) of the compounds all reach the nanomole level, and EC50 of some of the compounds reaches the picomole level, and therefore said compounds are excellent GLP-1 receptor agonist. In addition, the compounds have good druggability. Therefore, the selenium-containing compounds can be used for preparing GLP-1 receptor agonists, and are used for preparing drugs for preventing and / or treating GLP-1 receptor activation related diseases.

Description

Selenium-containing compound and use thereof Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a selenium-containing compound and a use thereof. Background Art

[0002] Selenium, a Period IV-Group VI element, has been less frequently used in medicinal chemistry and drug design. However, in recent years, researchers in this field have shown increasing interest in its application in drug design, development, and synthesis. Selenium, due to its large atomic structure and loose outer valence electrons, exhibits unique redox properties and nucleophilicity. Selenium-containing molecules typically exhibit higher bioreactivity than sulfur-containing molecules in vivo, readily forming Se-S adducts and significantly enhancing the selective uptake of cancer cells. Consequently, selenium-containing molecules are often used as antioxidants, free radical scavengers, or pro-oxidants in anti-inflammatory and anti-tumor therapies. Selenium atoms, as bioisosteres of oxygen atoms, have also been applied in drug design and natural product modification, resulting in numerous compounds acquiring unique pharmaceutical activities. The introduction or replacement of selenium atoms has become a novel strategy for improving compound properties.

[0003] GLP-1 is a rapidly metabolized endogenous incretin hormone produced in the intestine and central nervous system, acting on the GLP-1 receptor (GLP-1R). Intestinal GLP-1 is widely distributed throughout the body via the circulatory system, primarily regulating blood glucose levels and metabolism. Centrally produced GLP-1 functions as a neurotransmitter, suppressing appetite and food intake. Due to their crucial roles in regulating blood glucose and metabolism, GLP-1R agonists have become important agents for the treatment and management of metabolic diseases such as diabetes. Currently approved GLP-1R agonists are peptidomimetic compounds that mimic the GLP-1 peptide, such as exenatide, liraglutide, dulaglutide, and semaglutide. New types and formulations of GLP-1R agonists are currently being explored, including GLP-1 peptide analogs with iteratively optimized properties, orally available peptide agonists, small molecule agonists, and allosteric agonists.

[0004] Danuglipron (PF-06882961) is a small molecule direct GLP-1R agonist of the benzimidazole-6-carboxylic acid class developed by Pfizer, which has picomolar GLP-1R-cAMP signaling pathway agonist activity (EC 50 ) and full agonist effect (E maxDanuglipron is a GLP-1R partial agonist. Structural biology data indicate that the danuglipron compound occupies a portion of the GLP-1 peptide's active site, inducing and mimicking a conformation critical for ligand-receptor interactions through a structural water network. Danuglipron exhibits primate-specific agonist effects, with acceptable oral absorption and in vivo metabolism following oral administration. Glucose tolerance tests have shown that danuglipron can effectively improve glucose tolerance, promote insulin release, and effectively control blood sugar levels in experimental subjects. Phase II clinical trials for danuglipron's blood sugar control indication have concluded, and Phase III clinical trials are underway.

[0005] Currently, there are relatively few reports of small-molecule GLP-1R agonists. However, as the full-length structure of the GLP-1R before ligand binding is revealed, the structural types of small-molecule GLP-1R agonists will diversify. The reported molecule Danuglipron provides a template for this type of structural optimization, but its pharmacokinetic properties and toxicity are still not ideal.

[0006] Summary of the Invention

[0007] Purpose of the invention: The first purpose of the present invention is to provide a selenium-containing compound; the second purpose of the present invention is to provide the use of the selenium-containing compound, including use for preparing a GLP-1 receptor agonist and for preparing a drug for treating or alleviating a disease directly or indirectly by stimulating the GLP-1 receptor.

[0008] This invention has developed a novel selenium-containing, orally effective, small-molecule GLP-1R agonist based on the novel structure of danuglipron, which is used to prepare drugs for metabolic diseases that modulate GLP-1R-related signaling. Compared to peptide drugs, this small-molecule agonist is orally effective and improves the pharmacokinetic properties and cardiotoxicity of danuglipron.

[0009] Technical solution: The purpose of the present invention is achieved through the following technical solution:

[0010] The present invention provides a compound of formula I or a pharmaceutically acceptable salt, enantiomer, stereoisomer, hydrate, solvate or polymorph thereof:

[0011] in,

[0012] Ring A is selected from C 3-8 Cycloalkyl, C 2-9 Heterocyclic group, C 5-12 Spirobicyclic, C 5-12 Spiroheterobicyclic group, C 5-12 Fused bicyclic group, C 5-12Fused heterobicyclic group, C 5-12 Bridged ring or C 5-12 bridged heterocyclic group;

[0013] L is selected from -C(O)-, -O-, -S-, -CR 4 R 5 or -NR 6 -;

[0014] R 4 、R 5 are each independently selected from hydrogen, deuterium, halogen, cyano, C1-C6 alkyl, C1-C6 alkoxy, -NR 5a R 5b , 6-10 membered aryl, 5-8 membered heteroaryl, 3-8 membered saturated or partially saturated cycloalkyl, 3-8 membered saturated or partially saturated heterocyclic group; the C1-C6 alkyl or C1-C6 alkoxy is optionally substituted by one or more of the following substituents: halogen, oxo, cyano, hydroxyl, C3-C6 saturated or partially saturated cycloalkyl; the aryl, heteroaryl, saturated or partially saturated cycloalkyl, saturated or partially saturated heterocyclic group is optionally substituted by one or more of the following substituents: halogen, oxo, cyano or -NR 5a R 5b ;

[0015] R 5a 、R 5b Each is independently selected from hydrogen or C1-C6 alkyl;

[0016] R 6 Selected from hydrogen, C1-C6 alkyl, C3-C10 cycloalkyl, heterocyclic group, -S(O)2R 6a or -S(O)2N(R 6a )(NR 6b ); the cycloalkyl or heterocyclic group is optionally substituted by C1-C6 alkyl, -F or cyano;

[0017] R 6a 、R 6b Each is independently selected from -H, C1-C6 alkyl or C3-C10 cycloalkyl;

[0018] R 1C1-C6 alkyl substituted by 3-6 membered heterocyclyl, C1-C6 alkyl substituted by C3-C6 cycloalkyl, C1-C6 alkyl substituted by 5-8 membered heteroaryl, C1-C6 alkyl substituted by C1-C6 alkoxy, 3-6 membered heterocyclyl, C3-C6 cycloalkyl or 5-8 membered heteroaryl; the 3-6 membered heterocyclyl, C3-C6 cycloalkyl, 5-8 membered heteroaryl or C1-C6 alkoxy is optionally replaced by one of the following: substituted by one or more substituents: H, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 alkylamino, cyano, cyano-substituted C1-C6 alkyl, hydroxy, hydroxy-substituted C1-C6 alkyl, halo-substituted C1-C6 alkyl, halo-substituted C3-C6 cycloalkyl, halo-substituted C1-C6 alkoxy, halo-substituted C1-C6 alkylamino, acyl, amide, aminoacyl, sulfonyl or amino;

[0019] R 2 Each is independently selected from a hydrogen atom, deuterium, halogen, cyano, hydroxyl, oxo, C1-C6 alkyl, C1-C6 alkoxy, amino, 6-10 membered aryl, 5-8 membered heteroaryl, 3-8 membered saturated or partially saturated cycloalkyl, 3-8 membered saturated or partially saturated heterocyclic group;

[0020] R 3 Each is independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl; said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl being optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, cyano, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl;

[0021] X 1 、X 2 、X 3 and X 4 The same or different, and each independently selected from -CR 7 or N atoms;

[0022] R 7 is selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl;

[0023] G is selected from G1 or G2; the structural formulas of G1 and G2 are as follows:

[0024] Ring B and Ring C are each independently selected from a 6-12 membered aryl group or a 6-12 membered heteroaryl group;

[0025] Y and Z are each independently selected from -CH2-, -S-, -SO-, -SO2-, -(O=S=NH)-, -Se- or -Se(O)-;

[0026] R 8 、R 11 are the same or different and are each independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl; said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl being optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, cyano, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl;

[0027] R 9 、R 10 、R 12 、R 13 are the same or different and are each independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, said alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl being optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl;

[0028] EE is selected from -COOH or a carboxyl surrogate; the carboxyl surrogate is selected from:

[0029] m is 0, 1, 2, or 3;

[0030] n is 0, 1, 2, 3 or 4;

[0031] o is 0, 1, 2, 3, 4, or 5;

[0032] p is 0, 1, 2, 3, 4 or 5.

[0033] The present invention also provides a compound of formula II or a pharmaceutically acceptable salt, enantiomer, stereoisomer, hydrate, solvate or polymorph thereof:

[0034] in,

[0035] Ring A is selected from C 3-8 Cycloalkyl, C 2-9 Heterocyclic group, C 5-12 Spirobicyclic, C 5-12 Spiroheterobicyclic group, C5-12 Fused bicyclic group, C 5-12 Fused heterobicyclic group, C 5-12 Bridged ring group or C 5-12 bridged heterocyclic group;

[0036] L is selected from -C(O)-, -O-, -S-, -CR 4 R 5 or -NR 6 -;

[0037] R 4 、R 5 are each independently selected from hydrogen, deuterium, halogen, cyano, C1-C6 alkyl, C1-C6 alkoxy, -NR 5a R 5b , 6-10 membered aryl, 5-8 membered heteroaryl, 3-8 membered saturated or partially saturated cycloalkyl, 3-8 membered saturated or partially saturated heterocyclic group; the C1-C6 alkyl or C1-C6 alkoxy is optionally substituted by one or more of the following substituents: halogen, oxo, cyano, hydroxyl, C3-C6 saturated or partially saturated cycloalkyl; the aryl, heteroaryl, saturated or partially saturated cycloalkyl, saturated or partially saturated heterocyclic group is optionally substituted by one or more of the following substituents: halogen, oxo, cyano or -NR 5a R 5b ;

[0038] R 5a 、R 5b Each is independently selected from hydrogen or C1-C6 alkyl;

[0039] R 6 Selected from hydrogen, C1-C6 alkyl, C3-C10 cycloalkyl, heterocyclic group, -S(O)2R 6a or -S(O)2N(R 6a )(NR 6b ); the cycloalkyl or heterocyclic group is optionally substituted by C1-C6 alkyl, -F or cyano;

[0040] R 6a 、R 6b Each is independently selected from -H, C1-C6 alkyl or C3-C10 cycloalkyl;

[0041] R 1C1-C6 alkyl substituted by 3-6 membered heterocyclyl, C1-C6 alkyl substituted by C3-C6 cycloalkyl, C1-C6 alkyl substituted by 5-8 membered heteroaryl, C1-C6 alkyl substituted by C1-C6 alkoxy, 3-6 membered heterocyclyl, C3-C6 cycloalkyl or 5-8 membered heteroaryl; the 3-6 membered heterocyclyl, C3-C6 cycloalkyl, 5-8 membered heteroaryl or C1-C6 alkoxy is optionally replaced by one of the following: substituted by one or more substituents: H, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 alkylamino, cyano, cyano-substituted C1-C6 alkyl, hydroxy, hydroxy-substituted C1-C6 alkyl, halo-substituted C1-C6 alkyl, halo-substituted C3-C6 cycloalkyl, halo-substituted C1-C6 alkoxy, halo-substituted C1-C6 alkylamino, acyl, amide, aminoacyl, sulfonyl or amino;

[0042] R 2 Each is independently selected from a hydrogen atom, deuterium, halogen, cyano, hydroxyl, oxo, C1-C6 alkyl, C1-C6 alkoxy, amino, 6-10 membered aryl, 5-8 membered heteroaryl, 3-8 membered saturated or partially saturated cycloalkyl, 3-8 membered saturated or partially saturated heterocyclic group;

[0043] R 3 Each is independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl; said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl being optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, cyano, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl;

[0044] X 1 、X 2 、X 3 and X 4 The same or different, and each independently selected from -CR 7 or N atoms;

[0045] R 7 is selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl;

[0046] G is selected from G1 or G2; the structural formulas of G1 and G2 are as follows:

[0047] Ring B and Ring C are each independently selected from a 6-12 membered aryl group or a 6-12 membered heteroaryl group;

[0048] Y and Z are each independently selected from -CH2-, -S-, -SO-, -SO2-, -(O=S=NH)-, -Se- or -Se(O)-;

[0049] R 8 、R 11 are the same or different and are each independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl; said alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl being optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, cyano, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl;

[0050] R 9 、R 10 、R 12 、R 13 are the same or different and are each independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, said alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl being optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl;

[0051] R 14 each independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl, or heteroaryl, said alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl being optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl, or heteroaryl;

[0052] EE is selected from -COOH or a carboxyl surrogate; the carboxyl surrogate is selected from:

[0053] m is 0, 1, 2, or 3;

[0054] n is 0, 1, 2, 3 or 4;

[0055] o is 0, 1, 2, 3, 4, or 5;

[0056] p is 0, 1, 2, 3, 4, or 5;

[0057] q is 0, 1, 2, or 3.

[0058] In certain preferred embodiments,

[0059] Ring A is selected from C 3-8 Cycloalkyl or C 2-9 heterocyclic group;

[0060] L is selected from -CR 4 R 5 ;

[0061] R 1 C1-C6 alkyl substituted by 3-6 membered heterocyclyl, C1-C6 alkyl substituted by C3-C6 cycloalkyl, C1-C6 alkyl substituted by 5-8 membered heteroaryl, C1-C6 alkyl substituted by C1-C6 alkoxy;

[0062] EE is selected from -COOH or carboxyl surrogate;

[0063] R 4 、R 5 As defined in Formula I or Formula II.

[0064] In certain preferred embodiments,

[0065] Ring A is selected from C 2-9 heterocyclic group;

[0066] Y and Z are each independently selected from -Se- or -Se(O)-;

[0067] EE is selected from -COOH,

[0068] The compounds of the general formula I or II of the present invention are preferably the compounds shown in the following Table 1:

[0069] Table 1 Preferred compounds

[0070] The compounds of the general formula I or II described herein may also exist in the form of salts, hydrates, or solvates thereof, which are converted into compounds of the general formula I or II in vivo. For example, within the scope of the present invention, the compounds of the present invention may be converted into pharmaceutically acceptable salts according to processes known in the art and used in the form of salts.

[0071] Certain compounds of the present invention may exist in multiple crystalline forms.

[0072] All tautomeric forms of the compounds of Formula I or Formula II of the present invention are included within the scope of the present invention. The compounds of the present invention may exist in specific geometric or stereoisomeric forms. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups, and all such isomers and mixtures thereof are included within the scope of the present invention.

[0073] The present invention also provides a pharmaceutical composition comprising a compound of Formula I or Formula II or a pharmaceutically acceptable salt, enantiomer, stereoisomer, hydrate, solvate or polymorph thereof and a pharmaceutically acceptable carrier or excipient.

[0074] The pharmaceutical compositions of the present invention can be administered in various known ways, such as orally, parenterally, by inhalation spray, or via an implanted reservoir. The pharmaceutical compositions of the present invention can be administered alone or in combination with other drugs. Oral compositions can be in any orally acceptable dosage form, including but not limited to tablets, capsules, emulsions, suspensions, dispersions, and solutions. Commonly used pharmaceutically acceptable carriers or excipients include stabilizers, diluents, surfactants, lubricants, antioxidants, binders, colorants, fillers, emulsifiers, and the like.

[0075] Sterile injectable compositions can be formulated using suitable dispersing agents or wetting agents and suspending agents according to techniques known in the art. Pharmaceutically acceptable carriers and solvents that can be used include water, mannitol, sodium chloride solution, and the like.

[0076] Actual dosage levels of the active ingredients in the pharmaceutical compositions of the present invention may be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. The selected dosage level will depend upon a variety of factors, including the activity of the specific compound of the present invention or salt thereof employed, the route of administration, the time of administration, the rate of excretion of the specific composition employed, the duration of treatment, other drugs, compounds, and / or materials used in combination with the specific composition employed, the age, sex, weight, general health, and prior medical history of the patient being treated, and similar factors well known in the medical arts.

[0077] The present invention also provides the use of a compound of Formula I or Formula II or a pharmaceutically acceptable salt, enantiomer, stereoisomer, hydrate, solvate or polymorph thereof in the preparation of a GLP-1 receptor agonist.

[0078] The present invention also provides the use of a compound of Formula I or Formula II, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, hydrate, solvate, or polymorph thereof, in the preparation of a medicament for preventing and / or treating diseases associated with GLP-1 receptor stimulation. The medicament directly or indirectly treats or alleviates diseases by stimulating the GLP-1 receptor. Diseases associated with GLP-1 receptor stimulation include diabetes, diabetic complications, metabolic syndrome, obesity, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, Parkinson's disease, Alzheimer's disease, hypertension, hyperlipidemia, atherosclerosis, cardiovascular risk, coronary heart disease, or stroke. Beneficial effects:

[0079] The present invention provides a class of selenium-containing compounds, which have a complete GLP-1R-cAMP signaling pathway agonist effect (E max ), and the compound GLP-1R-cAMP half-maximal effect concentration (EC 50 ) have reached the nanomolar level, and the EC of some compounds 50 The selenium-containing compounds provided by the present invention have the potential to be developed into drugs for the prevention and / or treatment of diseases that are directly or indirectly treated or alleviated by stimulating the GLP-1 receptor. BRIEF DESCRIPTION OF THE DRAWINGS

[0080] FIG1 is a standard curve of the rHLM test of compound 1.

[0081] FIG2 is a standard curve of the rHLM test of PF-06882961.

[0082] Figure 3 is the PK test ct curve of PF-06882961.

[0083] FIG4 is a PK test ct curve of the low-dose group of compound 1.

[0084] FIG5 is a PK test ct curve of the high-dose group of compound 1. DETAILED DESCRIPTION

[0085] The following examples describe the essential contents of the present invention in detail, but are not intended to limit the scope of protection of the present invention. Experimental methods in the examples of the present invention where specific conditions are not specified are generally based on conventional conditions or conditions recommended by the raw material or product manufacturers; reagents where the source is not specified are generally commercially available conventional reagents.

[0086] Identification, characterization and purity determination methods of compounds:

[0087] 1. NMR: Compounds of the present invention1 H-NMR spectra were measured using a Bruker nuclear magnetic resonance spectrometer (300 MHz) and a Bruker nuclear magnetic resonance spectrometer (500 MHz). 13 C-NMR spectra were obtained using a Bruker nuclear magnetic resonance spectrometer (300 MHz). Chemical shifts (δ) are expressed in ppm using tetramethylsilane as the internal standard (0.00 ppm). 1 H-NMR notation: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of a doublet, dt = doublet of a triplet. Coupling constants, when given, are given in Hz.

[0088] 2. MS: The mass spectra of the compounds of the present invention were measured using a Shimadzu LC / MS-8050 triple quadrupole mass spectrometer using ESI as the ionization method. The molecular weights were accurate to one decimal place.

[0089] 3. HRMS: The high-resolution mass spectra of the compounds of the present invention were measured using LC / MS mass spectrometers from Shimadzu and Agilent. The testing method was Q-TOF, and the molecular weight was accurate to four decimal places.

[0090] 4. Purity determination: The purity of the compounds of the present invention was determined using a Shimadzu LC-20AT high performance liquid chromatograph. The test conditions were: Agilent XDS-C18 column (4.6 mm*250 mm), mobile phase: methanol: water (containing 0.1% formic acid) = 70:30, the test compound sample was a chromatographic methanol solution with a concentration of about 1 mg / mL, injection volume: 10 μL, measurement wavelength: 254 nm, detection time: 10 min. Purity was determined using retention time (t R ) and area ratio (%), accurate to two decimal places.

[0091] Example 1 Preparation of Compound 1

[0092] (S)-2-(4-(6-(4-cyano-2-fluorobenzyl)seleno)pyridin-2-yl)piperidin-1-methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 1)

[0093] Compound 1 was prepared using starting material 1-1 according to the following route.

[0094] Step 1: Preparation of 1-(tert-butyl)-4-methyl-4-(6-bromopyridin-2-yl)piperidine-1,4-dicarboxylic acid (Compound 1-3).

[0095] The raw materials, 2,6-dibromopyridine (compound 1-1, 1.6 g, 6.7 mmol) and methyl N-Boc-piperidine-4-carboxylate (compound 1-2, 1.75 g, 8.1 mmol), were dissolved in 3 mL of anhydrous THF and cooled to -78°C under N2 protection. 17 mL of a THF solution of LiHMDS (17 mmol) was slowly added. After reacting for 30 minutes, the mixture was warmed to room temperature and stirred for 2.5 hours. After completion of the reaction, dilute HCl solution was added dropwise to the reaction mixture to adjust the pH to neutral. The reaction mixture was extracted with EA, washed twice with saturated sodium chloride solution, and concentrated to yield a colorless oil, compound 1-3 (2.06 g, 77.1%). 1 H NMR(300MHz, DMSO-d6)δ7.78(t,J=7.8Hz,1H),7.59(dd,J=7.9,0.7Hz,1H),7.53-7.49(m,1H),3.70(d,J= 4.4Hz,1H),3.65(s,4H),3.08(s,2H),2.35-2.23(m,2H),1.97(ddd,J=13.9,7.2,3.2Hz,2H),1.41(s,9H). LC-MS:[M+H] + =398.1 / 400.1 (Br isotope).

[0096] Step 2: Preparation of 4-(6-bromopyridin-2-yl)-1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (Compound 1-4).

[0097] Intermediate 1-3 (2.06 g, 5.16 mmol) was dissolved in 20 mL of methanol, and a 2 M aqueous NaOH solution (26 mL) was added dropwise. The mixture was stirred at 60°C for 2 h. The reaction was complete after TLC. The mixture was acidified with dilute hydrochloric acid to a pH less than 6, extracted with EA, and concentrated to give white crystals, which were directly used in the next step without purification.

[0098] Step 3: Preparation of tert-butyl 4-(6-bromopyridin-2-yl)piperidine-1-carboxylate (Compound 1-5).

[0099] Intermediate 1-4 was dissolved in 45 mL of 1,2-dichloroethane (DCE) and heated to 95°C for 4 h. After TLC, the reaction was completed and dried under reduced pressure to obtain product 1-5 as a pale yellowish-white solid (1.52 g, 86.4%). 1H NMR (300MHz, DMSO-d6) δ7.70(t,J=7.7Hz,1H),7.49(d,J=7.8Hz,1H),7.37(d,J=7.6Hz,1H),4.07(d,J=13. 1Hz, 2H), 2.86 (tt, J=11.9, 2.8Hz, 3H), 1.83 (d, J=12.8Hz, 2H), 1.55 (td, J=12.5, 4.2Hz, 2H), 1.43 (s, 9H). LC-MS:[M+H] + =340.1 / 341.9.

[0100] Step 4: Preparation of tert-butyl 4-(6-selenylhydroxypyridin-2-yl)piperidine-1-carboxylate (Compound 1-6).

[0101] NaOH (22 mg, 0.53 mmol) and elemental selenium powder (42 mg, 0.53 mmol) were dissolved in 3 mL of anhydrous DMF. Under N2 protection, hydrazine hydrate (80%, 18 μL, 0.29 mmol) was added and the mixture was heated to 40°C and stirred for 2 h. A solution of intermediate 1-5 (100 mg, 0.29 mmol) in anhydrous DMF was added to the reaction system, and the mixture was heated to 160°C and stirred for 4 h. TLC indicated the reaction was complete. The reaction solution was filtered, extracted with equal volumes of water and EA, concentrated, and purified by column chromatography (eluent: petroleum ether:ethyl acetate = 30:1, v / v) to afford compound 1-6 (55 mg, 55.2%) as a yellow solid. 1 H NMR(300MHz,DMSO-d6)δ7.71–7.55(m,2H),7.24–7.09(m,1H),4.11–3.98(m,2H), 2.90–2.72(m,3H),1.84–1.73(m,2H),1.53(td,J=12.5,4.2Hz,2H),1.42(s,9H). LC-MS:[M+H] + =343.1.

[0102] Step 5: Preparation of tert-butyl 4-(6-((4-cyano-2-fluorobenzyl)selenyl)pyridin-2-yl)piperidine-1-carboxylate (Compound 1-8).

[0103] Intermediate 1-6 (411 mg, 1.20 mmol) was suspended in 10 mL of PEG-400, protected with N₂. NaBH₄ (136 mg, 3.60 mmol) was added and stirred at room temperature for 0.5 h. 4-Cyano-2-fluorobenzyl bromide (Compound 1-7, 257 mg, 1.20 mmol) was added and stirred for 15 min, resulting in the precipitation of a white insoluble material. TLC indicated the reaction was complete. The reaction solution was extracted with equal volumes of water and EA and purified by column chromatography (eluent: petroleum ether:ethyl acetate = 50:1, v / v) to afford Compound 1-8 (420 mg, 73.7%) as a colorless oil.

[0104] 1 H NMR(300MHz,Chloroform-d)δ7.66(t,J=7.7Hz,1H),7.59–7.54(m,1H),7.49(q,J=1.6Hz,1H),7.48–7.43(m,1H),7.28(d,J=8.1Hz,1H),7.07 (d,J=7.6Hz,1H),4.73–4.58(m,2H),4.52–4.27(m,2H),2.99(tt,J=12.0,4.1Hz,3H),2.06(d,J=12.9Hz,2H),1.98–1.88(m,2H),1.63(s,9H). LC-MS:[M+H] + =476.2.

[0105] Step 6: Preparation of 3-fluoro-4-((6-(piperidin-4-yl)pyridin-2-yl)selenyl)methyl)benzonitrile (Compound 1-9) p-toluenesulfonate.

[0106] Intermediate 1-8 (420 mg, 0.88 mmol) was dissolved in 10 mL of EA, and p-TSA·H2O (502 mg, 2.64 mmol) was added. The mixture was stirred and heated at 60°C. The reaction progress was monitored by TLC. After completion of the reaction, the solvent was evaporated to dryness to obtain crude compound 1-9, which was used directly in the next step without purification.

[0107] Step 7: Preparation of (S)-methyl 2-(4-(6-(4-cyano-2-fluorobenzyl)selenyl)pyridin-2-yl)piperidin-1-methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (Compound 1-11).

[0108] The crude intermediate 1-9 was dissolved in 10 mL of MeCN, and the starting material (S)-methyl 2-(chloromethyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylate (Compound 1-10, 235 mg, 0.80 mmol) and KCO (552 mg, 4.00 mmol) were added. The mixture was heated to 50°C with stirring. The reaction was monitored for completion by TLC. Purification by column chromatography (elution system: dichloromethane:methanol = 100:1, v / v) afforded Compound 1-11 (247 mg, 48.8%) as a pale white oil. 1 H NMR(300MHz,Chloroform-d)δ8.09(d,J=1.6Hz,1H),7.92–7.87(m,1H),7.70(d,J=7.7Hz,1H),7.43(t,J=7 .6Hz,1H),7.32(t,J=7.7Hz,1H),7.25(s,2H),7.04(d,J=7.8Hz,1H),6.86(d,J=7.7Hz,1H),5.14(d,J=6.0 Hz,1H),4.69–4.49(m,3H),4.44(d,J=1.4Hz,2H),4.32(dt,J=9.7,6.2Hz,1H),3.92(d,J=11.6Hz,2H),3.8 8(d,J=2.7Hz,3H),2.97(s,2H),2.78–2.59(m,2H),2.45–2.35(m,1H),2.24(s,2H),1.83(d,J=25.3Hz,4H). LC-MS:[M+H] + =634.2.

[0109] Step 8: Preparation of compound 1.

[0110] Intermediate 1-11 (247 mg, 0.39 mmol) was dissolved in 15 mL of MeCN, and a solution of TBD (114 mg, 0.82 mmol) in water was added. The mixture was stirred at room temperature overnight. TLC confirmed the reaction was complete, and the pH was adjusted to 5-6 using aqueous CA. Extraction with DCM and concentration were performed, followed by column chromatography (elution system: dichloromethane:methanol = 30:1, v / v) to afford Compound 1 (152 mg, 62.9%) as a dark white solid.

[0111] 1H NMR(300MHz,DMSO-d6)δ12.80(s,1H),8.28(s,1H),7.81(d,J=9.2Hz,2H),7.72–7.49( m,4H),7.29(d,J=7.9Hz,1H),7.08(d,J=7.5Hz,1H),5.11(s,1H),4.72(dd,J=37.1,11 .4 Hz, 2H), 4.52 (s, 2H), 4.40 (q, J = 8.9, 8.4 Hz, 2H), 3.90 (dd, J = 54.5, 13.8 Hz, 2H), 3.12–2.82 (m, 2H), 2.79–2.55 (m, 2H), 2.44 (s, 1H), 2.31–2.09 (m, 2H), 1.90–1.68 (m, 4H). HRMS (ESI): calculated value C 31 H 30 FN5O3Se, [M+H] + 620.1506, found 620.1556. HPLC: t R =2.89min, purity 99.56%.

[0112] Example 2 Preparation of Compound 2

[0113] (S)-2-(4-(6-Benzylselenyl)pyridin-2-yl)piperidin-1-methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 2)

[0114] Compound 2 was synthesized using benzyl bromide and intermediate 1-6 as starting materials according to the synthetic route of compound 1. Compound 2 was a white solid, 50.4 mg in total, with a yield of 62.5%. 1H NMR (300MHz, DMSO-d6) δ8.27(d,J=1.6Hz,1H),7.80(dd,J=8.5,1.6Hz,2H),7.76–7.69(m,2H),7.64(d,J=8.5Hz,1 H),7.60–7.50(m,3H),7.28(d,J=7.7Hz,1H),7.07(d,J=7.5Hz,1H),5.18–5.03(m,1H),4.86–4.60(m,2H),4.53(s , 2H), 4.36 (dt, J = 14.7, 6.3 Hz, 2H), 4.04–3.69 (m, 2H), 3.28 (d, J = 6.0 Hz, 1H), 3.04 (d, J = 10.9 Hz, 1H), 2.88 (d, J = 11.1 Hz, 1H), 2.65 (td, J = 17.0, 14.4, 6.4 Hz, 2H), 2.21 (dd, J = 22.5, 11.2 Hz, 2H), 1.86 (td, J = 13.0, 12.5, 6.7 Hz, 4H). HRMS (ESI): calculated value C 30 H 32 N4O3Se,[M+H] + 577.1712, found 577.1708. HPLC: t R =2.45min, purity 97.02%.

[0115] Example 3 Preparation of Compound 3

[0116] (S)-2-(4-(6-(4-cyanobenzyl)selenoyl)pyridin-2-yl)piperidin-1-methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 3)

[0117] Compound 3 was synthesized using 4-cyanobenzyl bromide and intermediate 1-6 as starting materials according to the synthetic route of compound 1. Compound 3 was a white solid, 46.7 mg in total, with a yield of 42.2%. 1H NMR(300MHz,DMSO-d6)δ12.83(s,1H),8.30(d,J=1.6Hz,1H),7.88–7.71(m,4H),7.67(d,J=8.4Hz,1H),7 .63–7.52(m,3H),7.31(d,J=7.8Hz,1H),7.10(d,J=7.6Hz,1H),5.18–5.04(m,1H),4.89–4.63(m,2H),4.5 6 (s, 2H), 4.41 (tq, J = 12.3, 5.6 Hz, 2H), 4.06–3.76 (m, 2H), 3.29 (d, J = 6.0 Hz, 1H), 3.07 (d, J = 11.1 Hz, 1H), 2.91 (d, J = 11.0 Hz, 1H), 2.80–2.60 (m, 2H), 2.24 (dd, J = 22.4, 11.1 Hz, 2H), 1.88 (dd, J = 15.5, 9.4 Hz, 4H). HRMS (ESI): calculated value C 31 H 31 N5O3Se,[M+H] + 602.1665, found 602.1651. HPLC: t R =3.15min, purity 98.92%.

[0118] Example 4 Preparation of Compound 4

[0119] (S)-2-(4-(6-(4-chlorobenzyl)seleno)pyridin-2-yl)piperidin-1-methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 4)

[0120] Compound 4 was synthesized using 4-chlorobenzyl bromide and intermediate 1-6 as starting materials according to the synthetic route of compound 1. Compound 4 was a white solid, 43.7 mg in total, with a yield of 47.1%. 1H NMR (300MHz, DMSO-d6) δ12.74(s,1H),8.27(s,1H),7.80(d,J=8.5Hz,1H),7.65(d,J=8.6Hz ,1H),7.52(t,J=7.7Hz,1H),7.40(d,J=8.1Hz,2H),7.28(dd,J=15.0,7.9Hz,3H),7.06(d,J= 7.6 Hz, 1H), 5.09 (d, J = 7.3 Hz, 1H), 4.86–4.58 (m, 2H), 4.54–4.29 (m, 4H), 4.09–3.72 (m, 2H), 3.27 (t, J = 6.0 Hz, 1H), 3.11–2.83 (m, 2H), 2.67 (s, 2H), 2.23 (d, J = 22.0 Hz, 2H), 1.84 (s, 4H). HRMS (ESI): calculated value C 30 H 31 ClN4O3Se,[M+H] + 611.1323, found 611.1317. HPLC: t R =5.39min, purity 96.75%.

[0121] Example 5 Preparation of Compound 5

[0122] (S)-2-(4-(6-(2-fluorobenzyl)selenoyl)pyridin-2-yl)piperidin-1-methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 5)

[0123] Compound 5 was synthesized using 2-fluorobenzyl bromide and intermediate 1-6 as starting materials according to the synthetic route of compound 1. Compound 5 was a white solid, 42.3 mg in total, with a yield of 38.3%. 1H NMR (300MHz, DMSO-d6) δ8.27(s,1H),7.85–7.75(m,1H),7.65(d,J=8.5Hz,1H),7.53(t,J=7.7Hz, 1H),7.45(t,J=7.5Hz,1H),7.25(dd,J=10.6,7.0Hz,2H),7.20–7.01(m,3H),5.10(d,J=7.7Hz,1H ), 4.88–4.57 (m, 2H), 4.48 (s, 2H), 4.43–4.31 (m, 2H), 4.09–3.71 (m, 2H), 2.96 (dd, J = 49.8, 11.0 Hz, 2H), 2.80–2.59 (m, 2H), 2.44 (d, J = 8.8 Hz, 1H), 2.35–2.11 (m, 2H), 1.85 (dt, J = 13.5, 6.5 Hz, 4H). HRMS (ESI): calculated value C 30 H 31 FN4O3Se,[M+H] + 595.1618, found 595.1617.HPLC: t R =6.51min, purity 94.38%.

[0124] Example 6 Preparation of Compound 6

[0125] (S)-2-(4-(6-(4-chloro-2-fluorobenzyl)selenoyl)pyridin-2-yl)piperidin-1-methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 6)

[0126] Compound 6 was synthesized using 4-chloro-2-fluorobenzyl bromide and intermediate 1-6 as starting materials according to the synthetic route of compound 1. Compound 6 was a white solid, 62.5 mg in total, with a yield of 75.9%. 1H NMR (300MHz, DMSO-d6) δ12.74(s,1H),8.27(s,1H),7.80(d,J=8.5Hz,1H),7.65(d,J=8.6Hz ,1H),7.52(t,J=7.7Hz,1H),7.40(d,J=8.1Hz,2H),7.28(dd,J=15.0,7.9Hz,3H),7.06(d,J= 7.6 Hz, 1H), 5.09 (d, J = 7.3 Hz, 1H), 4.86–4.58 (m, 2H), 4.54–4.29 (m, 4H), 4.09–3.72 (m, 2H), 3.27 (t, J = 6.0 Hz, 1H), 3.11–2.83 (m, 2H), 2.67 (s, 2H), 2.23 (d, J = 22.0 Hz, 2H), 1.84 (s, 4H). HRMS (ESI): calculated value C 30 H 30 ClFN4O3Se,[M+H] + 629.1256, found 629.1218. HPLC: t R =5.79min, purity 97.66%.

[0127] Example 7 Preparation of Compound 7

[0128] (S)-2-(4-(6-((2-methoxy-4-(trifluoromethyl)benzyl)selenoyl)pyridin-2-yl)piperidin-1-methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 7)

[0129] Compound 7 was synthesized using 2-methoxy-4-(trifluoromethyl)benzyl bromide and intermediate 1-6 as starting materials according to the synthetic route of compound 1. Compound 7 was a white solid, 45 mg in total, with a yield of 60.5%. 1H NMR(300MHz,DMSO-d6)δ8.26(s,1H),7.85–7.76(m,1H),7.64(d,J=8.5Hz,1H),7.52(dt,J=7.8,3.8Hz,2 H),7.29–7.15(m,3H),7.06(d,J=7.6Hz,1H),5.10(d,J=7.6Hz,1H),4.80(dd,J=15.0,7.2Hz,1H),4.71– 4.60 (m, 1H), 4.48–4.31 (m, 4H), 3.98 (d, J = 13.5 Hz, 1H), 3.88 (s, 3H), 3.79 (d, J = 13.4 Hz, 1H), 3.05 (d, J = 11.0 Hz, 1H), 2.90 (d, J = 11.1 Hz, 1H), 2.71 (s, 2H), 2.44 (d, J = 8.4 Hz, 1H), 2.48–2.14 (m, 2H), 1.85 (s, 4H). HRMS (ESI): calculated value C 32 H 33 F3N4O4Se,[M+H] + 675.1619, found 675.1657. HPLC: t R =7.22min, purity 96.63%.

[0130] Example 8 Preparation of Compound 8

[0131] (S)-2-(4-(6-(4-cyano-2-fluorobenzyl)seleno)pyridin-2-yl)piperazine-1-methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 8)

[0132] Compound 8 was prepared using starting materials 1-1 and 8-2 according to the following route.

[0133] Step 1: Preparation of tert-butyl 4-(6-bromopyridin-2-yl)piperazine-1-carboxylate (Compound 8-2).

[0134] The starting material, 2,6-dibromopyridine (compound 1-1, 346 mg, 1.46 mmol), was dissolved in 10 mL of DMF. 1-Boc-piperazine (compound 8-1, 353 mg, 1.89 mmol) and K2CO3 (405 mg, 2.93 mmol) were added, and the mixture was heated and stirred at 80°C for 4 h. TLC indicated the reaction was complete. Purification by column chromatography (elution system: petroleum ether:ethyl acetate = 20:1, v / v) afforded 8-2 (331 mg, 66.1%) as a white solid. 1H NMR (300MHz, Chloroform-d) δ7.33 (dd, J=8.4, 7.4Hz, 1H), 6.81 (d, J=7.5Hz, 1H), 6.56 (d, J=8.3Hz, 1H), 3.56 (s, 8H), 1.51 (s, 9H). LC-MS:[M+H] + =342.1.

[0135] Step 2: Preparation of tert-butyl 4-(6-selenylhydroxypyridin-2-yl)piperazine-1-carboxylate (Compound 8-3).

[0136] NaOH (180 mg, 4.50 mmol) and elemental selenium powder (356 mg, 4.50 mmol) were dissolved in 6 mL of anhydrous DMF. Under N₂ protection, hydrazine hydrate (80%, 150 μL, 3.0 mmol) was added and the mixture was heated to 40°C and stirred for 2 h. Intermediate 8-2 (514 mg, 1.50 mmol) was dissolved in 5 mL of anhydrous DMF and added to the reaction system. The mixture was heated to 160°C and stirred for 4 h. TLC indicated the reaction was complete. The reaction solution was filtered, extracted with equal volumes of water and EA, concentrated, and purified by column chromatography (eluent: petroleum ether:ethyl acetate = 15:1, v / v) to afford 8-3 (331 mg, 64.3%) as a yellow solid. 1 H NMR (300MHz, Chloroform-d) δ7.45 (t, J = 7.4 Hz, 1H), 7.12 (dd, J = 7.5, 1.5 Hz, 1H), 6.87 (dd, J = 7.5, 1.5 Hz, 1H), 6.19 (s, 1H), 3.62 (s, 8H), 1.46 (s, 9H). LC-MS:[M+H] + =344.0.

[0137] Step 3: Preparation of tert-butyl 4-(6-(4-cyano-2-fluorobenzyl)selenyl)pyridin-2-yl)piperazine-1-carboxylate (Compound 8-4).

[0138] Intermediate 8-3 (169 mg, 0.49 mmol) was suspended in 10 mL of PEG-400 under N₂ protection. NaBH₄ (56 mg, 0.49 mmol) was added and stirred at room temperature for 0.5 h. 4-Cyano-2-fluorobenzyl bromide (Compound 1-7, 105 mg, 0.49 mmol) was added and stirred for 15 min, resulting in the precipitation of a white insoluble material. TLC indicated the reaction was complete. The reaction solution was extracted with equal volumes of water and EA and purified by column chromatography (eluent: petroleum ether:ethyl acetate = 30:1, v / v) to afford compound 8-4 (128 mg, 73.7%) as a colorless oil. 1H NMR(300MHz,Chloroform-d)δ7.54(t,J=7.7Hz,1H),7.43–7.34(m,3H),6.72(d,J= 7.4Hz, 1H), 6.47 (d, J = 8.4Hz, 1H), 4.51 (d, J = 1.4Hz, 2H), 3.62 (s, 8H), 1.55 (s, 9H). LC-MS:[M+H] + =477.1.

[0139] Step 4: Preparation of compound 8.

[0140] According to the synthesis method of compound 1, intermediate 8-4 was used as the starting material to prepare compound 8. Compound 8 was a light yellow solid, 64.5 mg in total, with a yield of 43.7%. 1 H NMR(300MHz,DMSO-d6)δ8.28(d,J=1.6Hz,1H),7.86–7.77(m,2H),7.69–7.55(m,3H),7.35(t, J=7.9Hz,1H),6.69(d,J=7.4Hz,1H),6.59(d,J=8.4Hz,1H),5.11(d,J=7.3Hz,1H),4.81(dd,J= 15.4,7.2 Hz, 1H), 4.71–4.62 (m, 1H), 4.55–4.33 (m, 4H), 4.05–3.77 (m, 2H), 3.52 (d, J = 5.7 Hz, 4H), 2.70 (d, J = 9.2 Hz, 1H), 2.57 (d, J = 5.2 Hz, 4H), 2.42 (d, J = 9.0 Hz, 1H). HRMS (ESI): calculated value C 30 H 29 FN6O3Se,[M+H] + 621.1523, found 621.1523. HPLC: t R =5.00min, purity 97.48%.

[0141] Example 9 Preparation of Compound 9

[0142] (S)-2-(4-(6-(4-chloro-2-fluorobenzyl)selenoyl)pyridin-2-yl)piperazine-1-methyl)-1-(oxetane-2-methyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 9)

[0143] Compound 9 was synthesized from 4-chloro-2-fluorobenzyl bromide and intermediate 8-4 according to the synthetic route of compound 8. Compound 9 was a yellow solid, 79.4 mg in total, with a yield of 62.5%. 1 H NMR (300MHz, DMSO-d6) δ9.07(s,1H),8.22(s,1H),7.82(d,J=8.4Hz,1H),7.57(d,J=8.4Hz,1H),7.47–7.30( m,3H),7.20(dd,J=8.3,2.2Hz,1H),6.68(d,J=7.4Hz,1H),6.58(d,J=8.4Hz,1H),5.10(dt,J=9.4,4.7Hz,1H ), 4.72 (ddd, J = 40.5, 15.3, 5.1 Hz, 2H), 4.49 (q, J = 7.3 Hz, 1H), 4.38 (d, J = 9.1 Hz, 3H), 4.04–3.94 (m, 1H), 3.81 (d, J = 13.4 Hz, 1H), 3.27 (t, J = 5.9 Hz, 4H), 3.20 (t, J = 5.8 Hz, 4H), 2.77–2.66 (m, 1H), 2.44 (t, J = 9.2 Hz, 1H). HRMS (ESI): calculated value C 29 H 29 ClFN5O3Se,[M+H] + 630.1108, found 630.1180. HPLC: t R =4.99min, purity 94.42%.

[0144] Example 10 Preparation of Compound 10

[0145] 2-(4-(6-(4-cyano-2-fluorobenzyl)selensulfonyl)pyridin-2-yl)piperidin-1-methyl)-1-((S)-oxetan-2-yl)methyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 10)

[0146] Compound 10 was prepared from starting materials 1-9 according to the following route.

[0147] Step 1: Preparation of 3-fluoro-4-((6-(piperidin-4-yl)pyridin-2-yl)selensulfonyl)methyl)benzonitrile (Compound 10-1).

[0148] Crude intermediate 1-9 (108 mg, 0.29 mmol) was dissolved in 5 mL of DCM and 5 mL of MeOH and cooled to 0°C in an ice bath. NCS (46.5 mg, 0.35 mmol) was added and stirred for 0.5 h. To the mixture was added 5 mL of DCM and 10% aqueous NaOH (2.9 mL, 10 mL / mmol), followed by stirring for 5 min. TLC indicated completion of the reaction. Extraction was performed with equal volumes of water and DCM, followed by concentration and purification by column chromatography (elution system: dichloromethane:methanol = 50:1, v / v) to afford 10-1 (81 mg, 71.6%) as a colorless oil.

[0149] 1 H NMR(300MHz,Chloroform-d)δ7.72(t,J=7.7Hz,1H),7.39(ddd,J=17.5,7.8,1.3Hz,2H),7.27–7.14(m,3H),4 .35–4.12(m,2H),3.33–3.21(m,2H),2.84(m,J=22.2,14.8,11.9,3.2Hz,3H),1.99–1.71(m,4H), LC-MS: [M+H] + =392.0.

[0150] Step 2: Preparation of compound 10.

[0151] According to the synthesis method of compound 1, intermediate 10-1 was used as the starting material to prepare compound 10. Compound 10 was a white solid, 18.0 mg in total, with a yield of 40.5%. 1 H NMR (300MHz, DMSO-d6) δ9.07(s,1H),8.22(s,1H),7.82(d,J=8.4Hz,1H),7.57(d,J=8.4Hz,1H),7.47–7.30( m,3H),7.20(dd,J=8.3,2.2Hz,1H),6.68(d,J=7.4Hz,1H),6.58(d,J=8.4Hz,1H),5.10(dt,J=9.4,4.7Hz,1H ), 4.72 (ddd, J = 40.5, 15.3, 5.1 Hz, 2H), 4.49 (q, J = 7.3 Hz, 1H), 4.38 (d, J = 9.1 Hz, 3H), 4.04–3.94 (m, 1H), 3.81 (d, J = 13.4 Hz, 1H), 3.27 (t, J = 5.9 Hz, 4H), 3.20 (t, J = 5.8 Hz, 4H), 2.77–2.66 (m, 1H), 2.44 (t, J = 9.2 Hz, 1H). HRMS (ESI): calculated value C 31H 30 ClFN5O4Se,[M+H] + 630.1520, found 630.1579. HPLC: t R =4.99min, purity 94.42%.

[0152] Example 11 Preparation of Compound 11

[0153] (S)-2-(4-(6-(4-cyano-2-fluorophenyl)selenomethyl)pyridin-2-yl)piperidin-1-methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 11)

[0154] Compound 11 was prepared from starting material 11-1 according to the following route.

[0155] Step 1: Preparation of 3-fluoro-4-hydrogenselenylbenzonitrile (Compound 11-2).

[0156] NaOH (243 mg, 6.07 mmol) and elemental selenium powder (480 mg, 6.07 mmol) were dissolved in 5 mL of anhydrous DMF. Under N2 protection, hydrazine hydrate (80%, 128 μL, 2.02 mmol) was added and the mixture was heated to 40°C and stirred for 2 h. 3-Fluoro-4-iodobenzonitrile (compound 11-1, 500 mg, 2.02 mmol) was dissolved in 5 mL of anhydrous DMF and added to the reaction system. The mixture was heated to 160°C and stirred for 4 h. TLC indicated the reaction was complete. The reaction solution was filtered, extracted with equal volumes of water and EA, concentrated, and purified by column chromatography (eluent: petroleum ether:ethyl acetate = 20:1, v / v) to afford a yellow solid, compound 11-2 (410 mg, 50.9%). 1 H NMR (300MHz, Chloroform-d) δ7.71 (dd, J=8.1, 6.8Hz, 1H), 7.45–7.30 (m, 2H). LC-MS:[M+H] + =201.9.

[0157] Step 2: Preparation of tert-butyl 6-(hydroxymethyl)-3',6'-dihydro-(2,4'-bipyridine)-1'(2'H)-carboxylate (Compound 11-5).

[0158] 6-Chloro-2-hydroxymethylpyridine (compound 11-3, 231 mg, 1.62 mmol) was dissolved in 20 mL of dioxane and 2 mL of H₂O. N-Boc-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (compound 11-4, 999 mg, 3.23 mmol), Pd(dppf)Cl₂ (118 mg, 0.16 mmol), and Cs₂CO₃ (1153 mg, 3.55 mmol) were added. The mixture was heated to 90°C and stirred for 2 h under N₂ protection. TLC indicated the reaction was complete. The reaction solution was extracted with equal volumes of water and EA. The product was concentrated and purified by column chromatography (eluent: petroleum ether:ethyl acetate = 50:1, v / v) to afford compound 11-5 (369 mg, 78.7%) as a colorless oil. 1 H NMR(300MHz,Chloroform-d)δ7.67(t,J=7.7Hz,1H),7.11(d,J=7.7Hz,2H),6.56(s,1H),4.78(s,2H) ,4.31(s,2H),2.91(dd,J=12.2,4.3Hz,2H),1.99(s,2H),1.79(dd,J=12.6,4.4Hz,2H),1.54(s,9H). LC-MS: [M+H]+=291.3.

[0159] Step 3: Preparation of tert-butyl 4-(6-(hydroxymethyl)pyridin-2-yl)piperidine-1-carboxylate (Compound 11-6).

[0160] Intermediate 11-5 (369 mg, 1.27 mmol) was dissolved in 10 mL MeOH, and an equal amount of Pd / C (10%) was added, H2 was replaced, and stirred for 2 h. LC-MS / MS showed that the reaction was complete. The mixture was filtered and the solvent was dried to give a colorless oily liquid, namely compound 11-6 (369 mg, 99.2%). 1 HNMR(300MHz,Chloroform-d)δ7.67(t,J=7.7Hz,1H),7.11(d,J=7.7Hz,2H),4.78( s,2H),4.31(s,2H),2.91(m,4H),1.99(s,2H),1.79(d,J=12.6Hz,2H),1.54(s,9H). LC-MS: [M+H]+=293.3.

[0161] Step 4: Preparation of tert-butyl 4-(6-(bromomethyl)pyridin-2-yl)piperidine-1-carboxylate (Compound 11-7).

[0162] Intermediate 11-6 (369 mg, 1.27 mmol) was dissolved in 10 mL of DCM, and CBr (420 mg, 1.27 mmol) and PPh (499 mg, 1.90 mmol) were added. The mixture was stirred at room temperature for 3 h. TLC indicated the reaction was complete. Purification by column chromatography (eluent: petroleum ether:ethyl acetate = 20:1, v / v) afforded compound 11-7 (234 mg, 52.2%) as an off-white solid. 1 H NMR(300MHz,Chloroform-d)δ7.64(t,J=7.7Hz,1H),7.31–7.27(m,1H),7.07(d,J=7.8Hz,1H),4.53(s,2H),4. 25(d,J=13.1Hz,2H), 2.86(d,J=12.6Hz,3H), 1.92(d,J=13.1Hz,2H), 1.76–1.63(m,2H), 1.48(d,J=1.1Hz,9H). LC-MS: [M+H]+=355.1.

[0163] Step 5: Preparation of tert-butyl 4-(6-(4-cyano-2-fluorophenyl)selenomethyl)pyridin-2-yl)piperidine-1-carboxylate (Compound 11-8).

[0164] Intermediate 11-2 (66 mg, 0.33 mmol) and NaBH4 (37.4 mg, 0.99 mmol) were dissolved in 2 mL of anhydrous ethanol under N2 protection. A solution of Intermediate 11-7 (234 mg, 0.66 mmol) in ethanol (3 mL) was added dropwise and stirred at room temperature overnight. TLC indicated the reaction was complete. The reaction solution was extracted with equal volumes of water and EA and purified by column chromatography (eluting with petroleum ether:ethyl acetate = 30:1, v / v) to afford compound 11-8 (90 mg, 73.7%) as a colorless oil. 1 H NMR(300MHz,Chloroform-d)δ7.72(dt,J=8.1,5.7Hz,1H),7.56(td,J=10.4,9.1,5.3Hz,1H),7.37–7.22(m,2H),7.15(d,J=7.6Hz,1H),7 .01(d,J=7.7Hz,1H), 4.29(d,J=21.9Hz,4H), 2.82(d,J=12.6Hz,3H), 1.86(d,J=13.0Hz,2H), 1.69(dt,J=12.4,6.7Hz,2H), 1.49(s,9H). LC-MS: [M+H]+=476.1.

[0165] Step 6: Preparation of compound 11.

[0166] According to the synthesis method of compound 1, intermediate 11-8 was used as the starting material to prepare compound 10. Compound 10 was a dark white solid, 35 mg in total, with a yield of 67.4%. 1 H NMR(300MHz, DMSO-d6)δ13.12–12.50(m,1H),8.28(d,J=1.6Hz,1H),7.98–7.87(m,1H),7.86–7.74(m,2H),7.72–7.57(m,3H),7.30 (d,J=7.7Hz,1H),7.13(d,J=7.7Hz,1H),5.11(dt,J=9.3,4.7Hz,1H),4.83(dd,J=15.2,7.1Hz,1H),4.68(dd,J=15.1,2.8Hz,1H),4. 56–4.44 (m, 3H), 4.40 (dt, J = 9.0, 5.9 Hz, 1H), 3.97 (d, J = 13.5 Hz, 1H), 3.79 (d, J = 13.4 Hz, 1H), 3.01 (d, J = 11.0 Hz, 1H), 2.87 (d, J = 11.1 Hz, 1H), 2.80–2.68 (m, 1H), 2.60 (d, J = 11.1 Hz, 1H), 2.45 (dd, J = 9.9, 7.4 Hz, 1H), 2.30–2.10 (m, 2H), 1.70 (dd, J = 24.9, 11.4 Hz, 4H). HRMS (ESI): calculated value C 31 H 30 FN5O3Se,[M+H] + 620.1571, found 620.1531. HPLC: t R =3.26min, purity 98.20%.

[0167] Example 12 Preparation of Compound 12

[0168] (S)-2-(4-(6-(4-cyano-2-fluorobenzyl)selenoyl)pyridin-2-yl)piperidin-1-methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-ylhydroxylamine (Compound 12)

[0169] Compound 12 was prepared using compound 1 as a raw material according to the following route.

[0170] Step 1: Preparation of Intermediate 12-2

[0171] Compound 1 (70 mg, 0.11 mmol) was dissolved in 5 mL of anhydrous DMF. EDCI (65 mg, 0.34 mmol), HOBt (45.8 mg, 0.34 mmol), DIPEA (59 μL, 0.34 mmol), and NH2OTHP (compound 12-1, 26.5 mg, 0.22 mmol) were added and stirred at room temperature for 3 h. TLC confirmed the reaction was complete. The reaction solution was extracted with equal volumes of water and EA. The organic phases were combined and concentrated to afford crude intermediate 12-2 (58 mg, 48.8%). LC-MS: [M+H]+ = 719.2.

[0172] Step 2: Preparation of compound 12

[0173] Intermediate 12-2 (58 mg, 0.08 mmol) was dissolved in 5 mL of methanol, and a methanol solution of TFA (6.2 μL, 0.08 mmol) was added dropwise. The mixture was stirred at room temperature for 2 h. The reaction was confirmed to be complete by TLC. Compound 12 was purified by column chromatography (elution system: dichloromethane:methanol = 20:1, v / v) to obtain compound 12 as a white solid (36 mg, 70.3%). 1 H NMR (300MHz, DMSO-d6) δ11.17(s,1H),8.99(s,1H),8.08(s,1H),7.81(d,J=9.8Hz,1H),7.65(d,J=8.2Hz,1H),7.61(s, 3H),7.53(d,J=7.7Hz,1H),7.29(d,J=7.6Hz,1H),7.08(d,J=7.4Hz,1H),5.13(s,1H),4.76(t,J=11.0Hz,1H),4.62(d, J=14.9 Hz, 1H), 4.52 (s, 2H), 4.41 (q, J=6.9 Hz, 2H), 3.97 (d, J=13.4 Hz, 1H), 3.78 (d, J=13.5 Hz, 1H), 3.03 (d, J=11.0 Hz, 1H), 2.89 (d, J=11.0 Hz, 1H), 2.76–2.64 (m, 3H), 2.21 (dd, J=22.6, 11.5 Hz, 2H), 1.82 (d, J=11.9 Hz, 4H). HRMS (ESI): calculated value C 31 H 31 FN6O3Se,[M+H] + 635.1607, found 635.1650. HPLC: t R =2.51min, purity 99.60%.

[0174] Example 13 Preparation of Compound 13

[0175] (S)-2-(4-(6-(4-cyano-2-fluorobenzyl)selenoyl)pyridin-2-yl)piperidin-1-methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxamide methylsulfonamide (Compound 13)

[0176] Compound 13 was prepared using compound 1 as a raw material according to the following route.

[0177] Step 1: Preparation of compound 13.

[0178] Compound 1 (40 mg, 0.06 mmol) was dissolved in 5 mL of anhydrous DCM. 2-chloro-1-methylpyridinium iodide (33 mg, 0.13 mmol), DMAP (11.8 mg, 0.10 mmol), and Et3N (27 μL, 0.19 mmol) were added under ice-cooling. After stirring for 0.5 h, methanesulfonamide (compound 13-1, 6.8 mg, 0.07 mmol) was added and stirred at room temperature for 10 h. TLC confirmed the reaction was complete. The reaction solution was extracted with equal volumes of water and DCM. The organic phases were combined, concentrated, and purified by column chromatography (elution system: dichloromethane:methanol = 15:1, v / v) to afford compound 13, 27 mg, in a 60.0% yield. 1 H NMR (300MHz, DMSO-d6) δ8.35(d,J=1.6Hz,1H),7.83(t,J=8.8Hz,2H),7.73(d,J=8.5Hz,1H),7.70–7.52 (m,3H),7.32(d,J=7.8Hz,1H),7.11(d,J=7.6Hz,1H),5.12(d,J=7.1Hz,1H),4.80(dd,J=15.3,7.0Hz,1 H), 4.65 (d, J = 13.6 Hz, 1H), 4.50 (d, J = 21.6 Hz, 2H), 4.41 (dd, J = 9.2, 6.2 Hz, 3H), 2.86 (s, 1H), 2.79–2.64 (m, 2H), 2.41 (s, 1H), 2.27–2.12 (m, 2H), 1.98 (s, 3H), 1.94 (s, 2H), 1.47 (s, 1H), 1.34 (d, J = 4.7 Hz, 3H). HRMS (ESI): calculated value C 32 H 33 FN6O4SSe,[M+H] + 697.1433, found 697.1486. ​​HPLC: t R =2.90min, purity 93.43%.

[0179] Example 14 Preparation of Compound 14

[0180] (S)-2-(4-(6-(4-cyano-2-fluorobenzyl)selenoyl)pyridin-2-yl)piperidin-1-methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-methylsulfonylurea (Compound 14)

[0181] Compound 14 was prepared from starting material 14-1 according to the following route.

[0182] Step 1: Preparation of tert-butyl ((3-fluoro-4-nitrophenyl)sulfonyl)carbamate (Compound 14-2)

[0183] Dissolve tert-butyl carbamate (10.74 g, 91.7 mmol) in 100 mL of anhydrous acetonitrile and cool to -78°C under N2 protection. Dissolve 3-fluoro-4-nitrobenzenesulfonyl chloride (compound 14-1, 18.26 g, 76.4 mmol) in 150 mL of anhydrous acetonitrile and quickly add. After half an hour of reaction, warm to room temperature and stir for 1.5 hours. After completion of the reaction, add dilute HCl dropwise to the reaction mixture to adjust the pH to neutral, as determined by TLC. The reaction mixture is extracted three times with equal volumes of ethyl acetate and water. The organic layers are combined and concentrated under low pressure to remove the solvent. The residue is purified by column chromatography (eluting with petroleum ether:ethyl acetate = 3:1, v / v) to afford intermediate 14-2 (10.47 g, 42.8%).

[0184] 1 H NMR (300MHz, DMSO-d6) δ8.19 (dd, J = 8.8, 7.3Hz, 1H), 7.78–7.65 (m, 2H), 1.22 (s, 9H). LC-MS: [M+H]+=321.0.

[0185] Step 2: Preparation of (S)-tert-butyl((4-nitro-3-((oxetanin-2-ylmethyl)amino)phenyl)sulfonyl)carbamate (Compound 14-3).

[0186] Intermediate 14-2 (10.47 g, 32.71 mmol) was dissolved in 50 mL of acetonitrile, and DIPEA (6.83 ml, 39.25 mmol) and (S)-oxetan-2-ylmethylamine (3.42 g, 39.25 mmol) were added. The reaction was heated to 60°C for 4 h. After TLC, the reaction was quenched by adding saturated ammonium chloride solution. The reaction solution was extracted three times with equal volumes of ethyl acetate and water. The organic layers were combined and concentrated under low pressure to remove the solvent to afford Intermediate 14-3 (9.18 g, 72.5%). 1H NMR (300MHz, DMSO-d6) δ8.41(t,J=5.7Hz,1H),8.19(d,J=8.9Hz,1H),7.49(d,J=1.8Hz,1H),7.01(dd,J=9.0,1. 8Hz,1H),5.00(t,J=5.9Hz,1H),4.62–4.39(m,2H),3.72–3.57(m,3H),2.76–2.52(m,2H),1.26(d,J=9.3Hz,9H). LC-MS: [M+H]+=388.1.

[0187] Step 3: Preparation of (S)-tert-butyl((4-amino-3-((oxetanin-2-ylmethyl)amino)phenyl)sulfonyl)carbamate (Compound 14-4).

[0188] Intermediate 14-3 (9.18 g, 23.72 mmol) was dissolved in 40 mL of methanol, and 10% Pd / C (918 mg) was added. The reaction was heated to 50°C under N2 protection for 2 h. After completion of the reaction, the reaction solution was filtered through Celite and concentrated under low pressure to remove the solvent. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 100:1, v / v) to obtain Intermediate 14-4 (5.1 g, 60.2%).

[0189] 1 H NMR (300MHz, DMSO-d6) δ11.01(s,1H),6.99(dd,J=8.2,2.1Hz,1H),6.86(d,J=2.1Hz,1H),6.58(d,J=8.2Hz,1H),5.60(s,2H),4.95(dt,J=14 .1,6.0Hz,2H),4.61–4.41(m,2H),3.31(ddt,J=18.9,13.2,6.9Hz,2H),2.67(dtd,J=10.9,8.1,6.2Hz,1H),2.50–2.35(m,1H),1.30(s,9H). LC-MS: [M+H]+=358.1.

[0190] Step 4: Preparation of (S)-tert-butyl((2-(chloromethyl)-1-(oxetanin-2-ylmethyl)-1H-benzo[d]imidazol-6-yl)sulfonyl)carbamate (Compound 14-5).

[0191] Intermediate 14-4 (5.1 g, 14.24 mmol) was dissolved in 30 mL of acetonitrile, and p-toluenesulfonic acid hydrate (2.4 g, 1.42 mmol) and 2-chloro-1,1,1-trimethoxyethane (2.63 g, 17.09 mmol) were added. The reaction was heated to 60°C for 4 h. After completion of the reaction, the reaction solution was extracted three times with equal volumes of ethyl acetate and water. The organic layers were combined and concentrated under low pressure to remove the solvent. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 200:1, v / v) to obtain Intermediate 14-5 (2.4 g, 40.6%).

[0192] 1 H NMR (300MHz, DMSO-d6) δ11.64(s,1H),8.30(d,J=1.8Hz,1H),7.88(d,J=8.6Hz,1H) ,7.74(dd,J=8.6,1.8Hz,1H),5.21–5.09(m,2H),5.05(td,J=7.2,2.7Hz,1H),4.79( dd,J=15.5,6.9Hz,1H),4.67(dd,J=15.5,2.9Hz,1H),4.47(td,J=8.1,7.7,5.8Hz,1 H), 4.32 (dt, J = 9.1, 5.9 Hz, 1H), 2.79–2.61 (m, 1H), 2.46–2.29 (m, 1H), 1.27 (s, 9H). LC-MS: [M+H]+=416.1.

[0193] Step 5: Preparation of (S)-tert-butyl((2-((4-(6-((4-cyano-2-fluorobenzyl)selenyl)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetanin-2-ylmethyl)-1H-benzo[d]imidazol-6-yl)sulfonyl)carbamate (Compound 14-6).

[0194] Intermediate 1-9 (4.18 g, 5.82 mmol) was dissolved in 15 mL of acetonitrile, and K2CO3 (3.98 g, 28.85 mmol) was added. The mixture was stirred at room temperature for 0.5 h, followed by the addition of compound 14-5 (2.4 g, 5.77 mmol). The temperature was raised to 50°C and the reaction was allowed to react for 2 h. After completion of the reaction by TLC, the mixture was quenched by the addition of saturated ammonium chloride solution. The reaction solution was extracted three times with equal volumes of ethyl acetate and water. The organic layers were combined and concentrated under low pressure to remove the solvent. The residue was purified by column chromatography (eluting with dichloromethane:methanol = 50:1, v / v) to afford intermediate 14-6 (3 g, 68.9%).

[0195] 1H NMR (300MHz, DMSO-d6) δ11.58(s,1H),8.23(d,J=1.7Hz,1H),7.90–7.77(m,2H),7.76–7.49(m,4H),7.30(d,J=7.8Hz, 1H),7.08(d,J=7.6Hz,1H),5.78(s,1H),5.12(t,J=8.1Hz,1H),4.82(dd,J=15.3,7.1Hz,1H),4.69(d,J=15.0Hz,1H), 4.53(s,2H),4.47–4.30(m,2H),4.03(d,J=13.6Hz,1H),3.87(d,J=13.6Hz,1H),3.07(d,J=11.0Hz,1H),2.93(d,J=11 .1Hz,1H),2.69(s,2H),2.49–2.34(m,1H),2.27(dd,J=20.3,10.4Hz,2H),1.80(dd,J=24.2,11.6Hz,3H),1.27(s,9H). LC-MS: [M+H]+=755.2.

[0196] Step 6: Preparation of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)selenyl)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetanin-2-ylmethyl)-1H-benzo[d]imidazole-6-sulfonamide (Compound 14-7).

[0197] Intermediate 14-6 (3 g, 3.98 mmol) was dissolved in 20 mL of dichloromethane, and CF3COOH (4.54 g, 39.8 mmol) was added dropwise. The mixture was stirred at room temperature for 2 h. After completion of the reaction by TLC, saturated sodium bicarbonate solution was added to quench the reaction. The reaction mixture was extracted three times with equal volumes of ethyl acetate and water. The organic layers were combined and concentrated under low pressure to remove the solvent, yielding Intermediate 14-7 (2.18 g, 83.8%).

[0198] 1H NMR (300MHz, DMSO-d6) δ8.13(d,J=1.7Hz,1H),7.84–7.50(m,6H),7.29(t,J=3.8Hz,3H),7.08(d,J=7. 6Hz,1H),5.76(s,1H),5.11(d,J=6.8Hz,1H),4.79(dd,J=15.2,7.1Hz,1H),4.70–4.60(m,1H),4.52(s, 2H),4.45–4.34(m,2H),3.98(d,J=13.5Hz,1H),3.80(d,J=13.5Hz,1H),3.03(d,J=11.2Hz,1H),2.88(d ,J=11.2Hz,1H),2.68(s,2H),2.43(q,J=8.3Hz,1H),2.23(dt,J=21.4,11.5Hz,2H),1.88–1.73(m,3H). LC-MS: [M+H]+=655.1.

[0199] Step 7: Preparation of compound 14.

[0200] Intermediate 14-7 (500 mg, 0.76 mmol) was dissolved in 5 mL of acetonitrile, and phenyl N-methylcarbamate (127 mg, 0.84 mmol) and DBU (139 mg, 0.92 mmol) were added. The mixture was stirred at room temperature for 2 h. After TLC, the reaction was quenched by adding saturated ammonium chloride solution. The reaction solution was extracted three times with equal volumes of ethyl acetate and water. The organic layers were combined and concentrated under low pressure to remove the solvent. The residue was purified by column chromatography (eluting with dichloromethane:methanol = 20:1, v / v) to afford compound 14 (397 mg, 73.1%). 1H NMR(300MHz,DMSO-d6)δ10.69(s,1H), 8.25(d,J=1.7Hz,1H),7.87–7.76(m,2H),7.72(dd,J=8.6,1.7Hz,1H),7.63(d,J=5.9Hz,2H),7.55(t,J=7.7H z,1H),7.30(d,J=7.8Hz,1H),7.09(d,J=7.6Hz,1H),6.41(d,J=4.8Hz,1H),5.77(s,1H),5.14–5.06(m,1H),4. 81 (dd, J = 15.2, 7.0 Hz, 1H), 4.74–4.63 (m, 1H), 4.53 (s, 2H), 4.47–4.30 (m, 2H), 3.09 (s, 1H), 2.94 (s, 1H), 2.76–2.59 (m, 2H), 2.48 (s, 1H), 2.34–2.13 (m, 1H), 1.85 (s, 4H), 1.49 (s, 1H), 1.34 (s, 1H), 1.24 (d, J = 4.3 Hz, 3H). HRMS (ESI): calculated value C 32 H 34 FN7O5SSe,[M+H] + 712.1542, found 712.1582. HPLC: t R =3.60min, purity 94.67%.

[0201] Example 15 Preparation of Compound 15

[0202] (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)seleno)pyridin-2-yl)piperidin-1-yl)methyl)-N-(methylcarbamoyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-cyclopropylsulfonylurea (Compound 15)

[0203] Compound 15 was synthesized from cyclopropylcarbamic acid phenyl ester and intermediate 14-7 according to the synthetic route of compound 14. Compound 15 was a white solid, 62 mg in total, with a yield of 66.2%. 1H NMR(300MHz,DMSO-d6)δ10.71(s,1H),8.39–8.29(m,1H),7.98–7.74(m,3H),7.72–7.56(m,3H),7.36(d,J =7.9Hz,1H),7.13(d,J=7.5Hz,1H),7.02–6.96(m,1H),5.77(s,1H),5.04(d,J=7.0Hz,1H),4.89(s,2H),4 .75 (d, J = 15.2 Hz, 1H), 4.59–4.43 (m, 2H), 4.36 (d, J = 8.6 Hz, 1H), 3.83 (s, 2H), 3.04 (s, 1H), 2.74 (s, 1H), 2.37 (d, J = 7.4 Hz, 2H), 2.34–2.11 (m, 3H), 1.55–1.45 (m, 1H), 1.37–1.22 (m, 1H), 1.24 (s, 5H), 0.85 (m, 1H). HRMS (ESI): calculated value C 34 H 36 FN7O4SSe,[M+H] + 738.1771, found 738.1735. HPLC R =4.21min, purity 99.25%.

[0204] Example 16 Preparation of Compound 16

[0205] (S)-2-((4-(6-((2-fluoro-4-(trifluoromethyl)benzyl)selenoyl)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 16)

[0206] Compound 16 was synthesized using 1-(bromomethyl)-2-fluoro-4-(trifluoromethyl)benzene and intermediate 1-6 as starting materials according to the synthetic route of compound 1. Compound 16 was a white solid, 28.4 mg in total, with a yield of 59.9%. 1H NMR (300MHz, DMSO-d6) δ8.28(s,1H),7.82(d,J=8.4Hz,1H),7.78–7.54(m,2H),7.51(dd,J=11.8,7.9Hz,4H),7.32(dd,J=17.3,7.8 Hz,1H),7.09(dd,J=7.7,5.0Hz,1H),5.11(dd,J=7.2,2.8Hz,1H),4.81(dd,J=15.2,7.1Hz,1H),4.66(dd,J=15.1,3.0Hz,1H),4.54( d, J = 3.5 Hz, 3H), 4.49–4.30 (m, 1H), 4.11–3.93 (m, 1H), 3.81 (d, J = 13.4 Hz, 1H), 3.44–3.34 (m, 1H), 3.33–3.14 (m, 1H), 2.94–2.83 (m, 1H), 2.79–2.59 (m, 1H), 2.49–2.36 (m, 1H), 2.35–2.14 (m, 1H), 2.09–1.96 (m, 2H), 1.87 (dd, J = 11.2, 5.0 Hz, 2H), 1.85–1.66 (m, 1H). HRMS (ESI): calculated value C 31 H 30 F4N4O3Se,[M+H] + 663.1492, found 663.1467. HPLC R =4.17min, purity 95.77%.

[0207] Example 17 Preparation of Compound 17

[0208] (S)-2-((4-(6-((4-cyano-2-methoxybenzyl)seleno)pyridin-2-yl)piperidin-1-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 17)

[0209] Compound 17 was synthesized using 4-(bromomethyl)-3-methoxybenzonitrile and intermediate 1-6 as starting materials according to the synthetic route of compound 1. Compound 17 was a white solid, 42.3 mg in total, with a yield of 59.8%. 1H NMR (300MHz, DMSO-d6) δ12.79(s,1H),8.27(s,1H),7.81(d,J=8.4Hz,1H),7.65(d,J=8.4Hz ,1H),7.48(dd,J=14.3,6.7Hz,2H),7.28(dd,J=19.4,7.8Hz,2H),7.05(d,J=7.6Hz,1H),5.1 1(h,J=6.1Hz,1H),4.80(dd,J=15.2,7.1Hz,1H),4.65(dd,J=15.2,2.9Hz,1H),4.43(s,2H), 4.48–4.37(m,1H),4.35(dd,J=10.4,4.9Hz,1H),3.98(d,J=13.5Hz,1H),3.84(d,J=10.4Hz, 4H), 3.05 (d, J = 11.1Hz, 1H), 2.91 (d, J = 11.2Hz, 1H), 2.79–2.59 (m, 2H), 2.43 (s, 1H), 2.35–2.15 (m, 2H), 1.81 (dd, J = 23.9, 11.7Hz, 4H). HRMS (ESI): calculated value C 32 H 33 N5O4Se,[M+H] + 632.1771, found 632.1757. HPLC R =7.88min, purity 94.36%.

[0210] Example 18 Preparation of Compound 18

[0211] 2-((4-(6-((4-cyano-2-fluorobenzyl)seleno)pyridin-2-yl)piperidin-1-yl)methyl)-1-(2-methoxyethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 18)

[0212] Compound 18 was prepared from starting material 18-1 according to the following route.

[0213] Step 1: Preparation of methyl 3-((2-methoxyethyl)amino)-4-nitrobenzoate (18-3).

[0214] Compound 18-1 (1.90 g, 9.54 mmol) was dissolved in 20 mL of ACN, and compound 18-2 (1.00 mL, 11.45 mmol) and DIPEA (3.32 mL, 19.08 mmol) were added. The mixture was heated to 60°C and stirred for 8 h. After completion of the reaction, the mixture was extracted with water and EA. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification by column chromatography (elution system: PE:EA = 70:1, v / v) afforded 18-3 (1.72 g, 70.26% yield) as a yellow solid. 1 H NMR (300MHz, DMSO-d6) δ8.18(dd,J=9.1,4.1Hz,2H),7.57(d,J=1.7Hz,1H),7.15(dd,J=8.9,1.8Hz,1H),3.89(s,3H),3.66–3.50(m,4H),3.34(s,3H). LC-MS:[M+H] + =255.1.

[0215] Step 2: Preparation of methyl 4-amino-3-((2-methoxyethyl)amino)benzoate (18-4).

[0216] Intermediate 18-3 (1.72 g, 6.69 mmol) was dissolved in 20 mL of THF, and 10% Pd / C (170 mg) was added. The mixture was stirred at room temperature for 2 h under a hydrogen atmosphere. After completion of the reaction, the reaction mixture was filtered through celite, washed with DCM, and the filtrate was concentrated under reduced pressure. Purification by column chromatography (elution system: PE:EA = 10:1, v / v) afforded 18-4 (1.32 g, 86.73% yield) as a colorless oil.

[0217] 1 H NMR (300MHz, DMSO-d6) δ7.17(dd,J=8.1,1.9Hz,1H),6.99(d,J=1.9Hz,1H),6.56(d,J=8.1Hz,1H),5.46 (s, 2H), 4.65 (t, J = 5.5Hz, 1H), 3.73 (s, 3H), 3.56 (t, J = 5.6Hz, 2H), 3.30 (s, 3H), 3.22 (q, J = 5.6Hz, 2H). LC-MS:[M+H] + =225.1.

[0218] Step 3: Preparation of methyl 2-(chloromethyl)-1-(2-methoxyethyl)-1H-benzo[d]imidazole-6-carboxylate (18-5).

[0219] Intermediate 18-4 (1.32 g, 5.80 mmol) was dissolved in 20 mL of ACN, and p-TSA·H2O (220.02 mg, 1.16 mmol) and 2-chloro-1,1,1-trimethoxyethane (1.56 mL, 11.60 mmol) were added. The mixture was heated to 60°C and stirred for 4 h. After TLC, the reaction was completed, extracted with water and EA. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification by column chromatography (elution system: PE:EA = 20:1, v / v) afforded a colorless oil (989.27 mg, 59.97% yield).

[0220] 1 H NMR (300MHz, DMSO-d6) δ8.29(dd,J=1.7,0.7Hz,1H),7.87(dd,J=8.5,1.6Hz,1H),7.73(dd,J=8.5 ,0.6Hz,1H),5.10(s,2H),4.61(t,J=5.1Hz,2H),3.90(s,3H),3.70(t,J=5.1Hz,2H),3.21(s,3H). LC-MS:[M+H] + =283.1.

[0221] Step 4: Preparation of methyl 2-((4-(6-((4-cyano-2-fluorobenzyl)selenyl)pyridin-2-yl)piperidin-1-yl)methyl)-1-(2-methoxyethyl)-1H-benzo[d]imidazole-6-carboxylate (18-6).

[0222] Using intermediate 18-5 and intermediate 1-6 as raw materials, the synthetic route of compound 1 was used to obtain white solid 18-6 (307 mg, yield 66.8%).

[0223] Step 5: Preparation of 2-((4-(6-((4-cyano-2-fluorobenzyl)selenyl)pyridin-2-yl)piperidin-1-yl)methyl)-1-(2-methoxyethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 18).

[0224] Compound 18 was synthesized using intermediate 18-6 as the starting material according to the synthetic route of compound 1. Compound 18 was a white solid, 265.28 mg in total, with a yield of 66.1%. 1H NMR (300 MHz, DMSO-d6) δ 12.81 (s, 1H), 8.19 (s, 1H), 7.81 (d, J = 8.7 Hz, 2H), 7.68–7.58 (m, 3H), 7.52 (d, J = 9.9 Hz, 1H), 7.28 (d, J = 7.9 Hz, 1H), 7.07 (d, J = 7.4 Hz, 1H), 4.62 (s, 2H), 4.50 (s, 2H), 3.87 (s, 2H), 3.75 (s, 2H), 3.52 (s, 1H), 2.95 (d, J = 10.6 Hz, 2H), 2.71 (s, 1H), 2.24 (s, 2H), 1.82 (s, 5H). HRMS (ESI): calculated value C 30 H 30 FN5O3Se,[M+H] + 608.1571, found 608.1551; HPLC R =5.85min, purity 97.32%.

[0225] Example 19 Preparation of Compound 19

[0226] 2-((4-(6-((4-cyano-2-fluorobenzyl)seleno)pyridin-2-yl)piperidin-1-yl)methyl)-1-(tetrahydrofuran-3-yl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 19)

[0227] Compound 19 was synthesized using 3-aminotetrahydrofuran and compound 18-1 as starting materials according to the synthetic route of compound 18. Compound 19 was a white solid, with a total amount of 193.39 mg and a yield of 72.5%. 1 H NMR (300MHz, DMSO-d6) δ12.79(s,1H),8.34(s,1H),7.81(d,J=9.1Hz,2H),7.71–7.48(m,4H),7.28(d ,J=7.8Hz,1H),7.07(d,J=7.5Hz,1H),5.67(s,1H),4.52(s,2H),4.28–4.17(m,1H),4.11(dd,J=10.5 , 3.0 Hz, 1H), 4.02–3.89 (m, 2H), 3.84 (d, J = 13.4 Hz, 1H), 3.64 (q, J = 8.7 Hz, 1H), 2.95 (d, J = 10.9 Hz, 1H), 2.84 (d, J = 10.9 Hz, 1H), 2.71 (s, 1H), 2.20 (q, J = 11.9 Hz, 3H), 1.82 (s, 3H), 1.74 (d, J = 11.7 Hz, 2H). HRMS (ESI): calculated value C 31H 30 FN5O3Se,[M+H] + 620.1571, found 620.1551. HPLC R =4.16min, purity 97.96%.

[0228] Example 20 Preparation of Compound 20

[0229] 2-((4-(6-((4-cyano-2-fluorobenzyl)seleno)pyridin-2-yl)piperidin-1-yl)methyl)-1-((1-ethyl-1H-imidazol-5-yl)methyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 20)

[0230] Compound 20 was synthesized using (1-ethyl-1H-imidazol-5-yl)methanamine and compound 18-1 as starting materials according to the synthetic route of compound 18. Compound 20 was a white solid, 64.92 mg in total, with a yield of 49.3%. 1 H NMR (300 MHz, DMSO-d6) δ 12.70 (s, 1H), 8.07 (s, 1H), 7.86–7.76 (m, 3H), 7.72–7.48 (m, 4H), 7.28 (d, J = 7.8 Hz, 1H), 7.02 (d, J = 7.6 Hz, 1H), 6.44 (s, 1H), 4.50 (s, 2H), 4.07–3.92 (m, 2H), 3.84 (s, 2H), 3.27 (t, J = 6.0 Hz, 3H), 3.24–3.13 (m, 3H), 2.92 (d, J = 10.9 Hz, 2H), 2.18 (m, 2H), 1.88 (dd, J = 12.0, 6.0 Hz, 4H). HRMS (ESI): calculated value C 31 H 30 FN5O3Se,[M+H] + 658.1839, found 658.1834. HPLC R =7.92min, purity 94.66%.

[0231] Example 21 Preparation of Compound 21

[0232] 2-((4-(6-((4-cyano-2-fluorobenzyl)seleno)pyridin-2-yl)piperidin-1-yl)methyl)-1-(thiazol-5-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 21)

[0233] Compound 21 was synthesized using thiazole-5-methylamine and compound 18-1 as starting materials according to the synthetic route of compound 18. Compound 21 was a white solid, 58.72 mg in total, with a yield of 55.6%. 1 H NMR (300 MHz, DMSO-d6) δ 8.86 (s, 1H), 8.10 (s, 1H), 7.96 (s, 1H), 7.80–7.67 (m, 2H), 7.60 (d, J = 8.4 Hz, 1H), 7.52 (s, 1H), 7.47 (t, J = 7.7 Hz, 2H), 7.21 (d, J = 7.8 Hz, 1H), 6.98 (d, J = 7.6 Hz, 1H), 5.90 (s, 2H), 4.44 (s, 2H), 3.83 (s, 2H), 2.94 (d, J = 10.9 Hz, 2H), 2.63 (m, 1H), 2.18 (m, 2H), 1.73 (s, 3H), 1.73–1.59 (m, 1H). HRMS (ESI): calculated value C 31 H 27 FN6O2SSe,[M+H] + 646.1138, found 646.1132. HPLC R =4.81min, purity 97.65%.

[0234] Example 22 Preparation of Compound 22

[0235] (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)seleno)pyridin-2-yl)-5,6-dihydro-1,2,4-triazin-1(4H)-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 22)

[0236] Compound 22 was prepared from starting material 22-1 according to the following route.

[0237] Step 1: Preparation of tert-butyl (6-bromopyridin-2-yl)carbamate (22-3).

[0238] Compound 22-1 (10.00 g, 36.76 mmol) was dissolved in 150 mL of DMF, and Cs2CO3 (35.91 g, 110.29 mmol) was added. The mixture was stirred at room temperature for 0.5 h, and compound 22-2 (9.50 mL, 110.29 mmol) was added. The mixture was heated to 70°C and stirred for 3 h. After completion of the reaction, the mixture was extracted with water and EA. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification by column chromatography (elution system: PE:EA = 200:1, v / v) afforded 22-3 (10.51 g, 91.25% yield) as a colorless oil.

[0239] 1 H NMR(300MHz,DMSO-d6)δ7.77–7.63(m,2H),7.36(dd,J=6.3,2.0Hz,1H),5.9 8–5.79(m,1H),5.17–5.03(m,2H),4.43(dt,J=5.4,1.6Hz,2H),1.46(s,9H). LC-MS:[M+H] + =313.1.

[0240] Step 2: Preparation of tert-butyl (6-bromopyridin-2-yl)(2-oxoethyl)carbamate (22-4).

[0241] Intermediate 22-3 (10.51 g, 33.52 mmol) was dissolved in 200 mL of a 1:1 (v / v) mixture of THF:H₂O. Potassium osmate dihydrate (105.05 mg, 0.33 mmol) and NaIO₄ (36.25 g, 167.62 mmol) were added and stirred at room temperature for 1 h. A large amount of white solid precipitated. After completion of the reaction, the mixture was added with saturated sodium thiosulfate solution and stirred for 10 min. The mixture was then extracted with EA. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification by column chromatography (eluting with PE:EA = 100:1, v / v) afforded 22-4 (9.75 g, 92.65% yield) as a colorless oil. 1 H NMR (300MHz, DMSO-d6) δ9.63 (s, 1H), 7.82 (d, J = 8.2 Hz, 1H), 7.74 (t, J = 7.9 Hz, 1H), 7.37 (d, J = 7.4 Hz, 1H), 4.68 (s, 2H), 1.45 (s, 9H). LC-MS:[M+H] + =315.1.

[0242] Step 3: Preparation of tert-butyl 2-(2-((6-bromopyridin-2-yl)(tert-butoxycarbonyl)amino)ethyl)hydrazine-1-carboxylate (22-6).

[0243] Intermediate 22-4 (9.75 g, 30.94 mmol) was dissolved in 150.00 mL of DCE, and compound 22-5 (8.23 g, 61.87 mmol) was added. The mixture was stirred at room temperature for 1 h. After TLC, NaBH3CN (5.84 g, 92.81 mmol) and NaBH(OAc)3 (19.78 g, 92.81 mmol) were added. After stirring at room temperature for approximately 3 h, the reaction was quenched with saturated aqueous NaHCO3 and extracted with DCM. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification by column chromatography (elution system: PE:EA = 10:1, v / v) gave a colorless oily liquid (6.86 g, yield 50.79%). 1 H NMR (300MHz, DMSO-d6) δ8.13(s,1H),7.76–7.66(m,1H),7.62(dd,J=8.2,0.9Hz,1H),7.37(dd,J=7. 5,0.9Hz,1H),4.58(s,1H),3.83(t,J=7.2Hz,2H),2.87(t,J=7.2Hz,2H),1.46(s,9H),1.38(s,9H). LC-MS:[M+H] + =431.1.

[0244] Step 4: Preparation of 6-bromo-N-(2-hydrazineethyl)pyridin-2-amine (22-7).

[0245] Intermediate 22-6 (6.86 g, 15.76 mmol) was dissolved in 120 mL of DCM. 60 mL of a 4 M solution of hydrogen chloride in dioxane was added. The mixture was stirred at room temperature for approximately 5 h. A large amount of white solid precipitated. After TLC analysis, the solvent was evaporated under reduced pressure. The resulting hydrochloride was directly used in the next reaction without purification. LC-MS: [M+H] + =231.1.

[0246] Step 5: Preparation of 6-bromo-N-(2-hydrazineethyl)pyridin-2-amine (22-8).

[0247] The hydrochloride product from the previous step was dissolved in 80 mL of glacial acetic acid, and 40 mL of trimethyl orthoformate was added. The mixture was heated to 100°C and stirred for 12 h under N₂ protection. After completion of the reaction, the solvent was evaporated, and saturated aqueous NaHCO₃ was added. The mixture was extracted with EA. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification by column chromatography (elution system: PE:EA = 2:1, v / v) afforded 22-8 as a white solid (4.18 g, 91.85% yield). 1 H NMR (300MHz, DMSO-d6) δ8.49(s,1H),7.87(d,J=13.9Hz,1H),7.72(m,1H),7.27(m,2H),3.94–3.80(m,2H),3.85(s,2H). LC-MS:[M+H] + =268.9.

[0248] Step 6: Preparation of 4-(6-bromopyridin-2-yl)-1,4,5,6-tetrahydro-1,2,4-triazine (22-9).

[0249] Intermediate 22-8 (4.18 g, 15.29 mmol) was dissolved in 100 mL of a 1:1 (v / v) mixture of DCM:MeOH. In an ice bath (0°C), 15.28 mL of 4M aqueous hydrochloric acid was added dropwise, and the mixture was stirred at room temperature overnight. After completion of the reaction, TLC analysis was performed, and a large amount of saturated aqueous NaHCO₃ was added. The mixture was extracted with DCM, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification by column chromatography (elution system: PE:EA = 1:2, v / v) afforded 22-9 (1.62 g, 43.67% yield) as a yellow solid. 1 H NMR (300MHz, DMSO-d6) δ7.75 (s, 1H), 7.62 (t, J = 8.0 Hz, 1H), 7.08 (dd, J = 16.4, 7.9 Hz, 2H), 6.35 (s, 1H), 3.73 (t, J = 5.0 Hz, 2H), 3.12 (t, J = 5.1 Hz, 2H). LC-MS:[M+H] + =241.0.

[0250] Step 7: Preparation of tert-butyl 4-(6-bromopyridin-2-yl)-5,6-dihydro-1,2,4-triazine-1(4H)-carboxylate (22-10).

[0251] Intermediate 22-9 (1.62 g, 6.66 mmol) was dissolved in 30 mL of DCM, and DMAP (81.42 mg, 0.66 mmol), TEA (1.15 mL, 8.00 mmol), and Boc2O (3.18 mL, 13.32 mmol) were added. The mixture was stirred at room temperature overnight. After TLC, the reaction was extracted with water and DCM. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Purification by column chromatography (elution system: PE:EA = 4:1, v / v) afforded 22-10 (780.03 mg, 34.47% yield) as a white solid.

[0252] 1 H NMR (300MHz, DMSO-d6) δ7.80 (s, 1H), 7.70 (t, J = 7.9 Hz, 1H), 7.22 (dd, J = 8.0, 5.1 Hz, 2H), 3.82 (tt, J = 4.6, 2.7 Hz, 4H), 1.46 (s, 9H). LC-MS:[M+H] + =341.0.

[0253] Step 8: Preparation of (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)selenyl)pyridin-2-yl)-5,6-dihydro-1,2,4-triazin-1(4H)-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 22).

[0254] Compound 22 was synthesized from intermediate 22-10 according to the synthetic route of compound 1. Compound 22 was a white solid, 45.84 mg in total, with a yield of 75.2%. 1 H NMR(300MHz,DMSO-d6)δ12.77(s,1H),8.28(s,1H),7.95(s,1H),7.84–7.77(m,2H ),7.68–7.51(m,4H),7.02(d,J=7.5Hz,1H),6.84(d,J=8.3Hz,1H),5.10–5.01(m,1 H),4.83(dd,J=15.5,7.1Hz,1H),4.70(d,J=14.1Hz,1H),4.56–4.30(m,6H),3.85( s, 2H), 3.14 (d, J = 5.2Hz, 2H), 2.69 (dd, J = 16.5, 4.7Hz, 1H), 2.39 (t, J = 9.4Hz, 1H). HRMS (ESI): calculated value C 29 H 26 FN7O3Se,[M+H] +620.1319, found 620.1325. HPLC R =4.50min, purity 98.18%.

[0255] Example 23 Preparation of Compound 23

[0256] (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)seleno)pyridin-2-yl)piperidin-1-yl)methyl)-4-fluoro-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 23)

[0257] Compound 23 was synthesized from methyl 3,5-difluoro-4-nitrobenzoate and (S)-oxetane-2-methylamine according to the synthetic route of compound 18. Compound 23 was a white solid, 52.25 mg in total, with a yield of 60.8%. 1 H NMR (300MHz, DMSO-d6) δ13.07(s,1H),8.15(s,1H),7.81(d,J=9.9Hz,1H),7.67–7.46(m,4H),7. 29(d,J=7.8Hz,1H),7.08(d,J=7.6Hz,1H),5.15–5.06(m,1H),4.84(d,J=16.0Hz,1H),4.68(d,J =14.8 Hz, 1H), 4.52 (s, 2H), 4.39 (q, J = 8.2 Hz, 3H), 3.99 (d, J = 13.8 Hz, 1H), 3.81 (d, J = 13.8 Hz, 1H), 3.03 (d, J = 10.1 Hz, 1H), 2.88 (d, J = 11.1 Hz, 1H), 2.69 (s, 3H), 2.26 (s, 1H), 1.82–1.77 (m, 4H). HRMS (ESI): calculated value C 31 H 29 F2N5O3Se,[M+H] + 638.1476, found 628.1463. HPLC t R =4.36min, purity 98.19%.

[0258] Example 24 Preparation of Compound 24

[0259] (S)-2-((4-(6-((4-cyano-2-fluorobenzyl)seleno)pyridin-2-yl)-5,6-dihydro-1,2,4-triazin-1(4H)-yl)methyl)-4-fluoro-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 24)

[0260] Compound 24 was synthesized using intermediate 22-10, intermediate 1-7, and methyl 3,5-difluoro-4-nitrobenzoate as raw materials according to the synthetic route of compound 22. Compound 24 was a white solid, 39.82 mg in total, with a yield of 57.9%. 1 H NMR (300 MHz, DMSO-d6) δ 13.11 (s, 1H), 8.17 (s, 1H), 7.96 (s, 1H), 7.81 (d, J = 10.0 Hz, 1H), 7.63–7.47 (m, 4H), 7.03 (d, J = 7.6 Hz, 1H), 6.84 (d, J = 8.0 Hz, 1H), 5.07 (d, J = 8.2 Hz, 1H), 4.88 (d, J = 15.1 Hz, 1H), 4.73 (d, J = 15.2 Hz, 1H), 4.47 (t, J = 10.8 Hz, 5H), 4.37 (s, 1H), 3.86 (s, 2H), 3.17 (s, 2H), 2.69 (s, 1H), 2.39 (s, 1H). HRMS (ESI): calculated value C 29 H 25 F2N7O3Se, [M+H] + 638.1225, found 638.1236. HPLC R =5.84min, purity 96.71%.

[0261] Example 25 Preparation of Compound 25

[0262] (S)-4-Fluoro-2-((4-(6-((2-methoxy-4-(trifluoromethyl)benzyl)seleno)pyridin-2-yl)-5,6-dihydro-1,2,4-triazin-1(4H)-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 25)

[0263] Compound 25 was synthesized using intermediate 22-10, 2-methoxy-4-(trifluoromethyl)benzyl bromide, and methyl 3,5-difluoro-4-nitrobenzoate as starting materials according to the synthetic route of compound 22. Compound 25 was a white solid (45.07 mg) with a yield of 60.4%.

[0264] 1H NMR(300MHz,DMSO-d6)δ13.11(s,1H),8.18(s,1H),7.98(s,1H),7.58–7.44(m,3H),7. 27–7.16(m,2H),7.00(d,J=7.6Hz,1H),6.84(dd,J=14.7,8.4Hz,1H),5.07(d,J=6.9Hz, 1H), 4.88 (dd, J = 15.5, 7.0 Hz, 1H), 4.80–4.68 (m, 1H), 4.59–4.28 (m, 6H), 3.88 (s, 5H), 3.37 (s, 1H), 3.17 (d, J = 6.5 Hz, 1H), 2.67 (d, J = 8.6 Hz, 1H), 2.41 (dd, J = 16.7, 8.0 Hz, 1H). HRMS (ESI): calculated value C 30 H 28 F4N6O4Se,[M+H] + 693.1346, found 693.1344. HPLC R =4.81min, purity 98.68%.

[0265] Example 26 Preparation of Compound 26

[0266] (S)-2-((4-(6-((4-cyano-2-methoxybenzyl)seleno)pyridin-2-yl)-5,6-dihydro-1,2,4-triazin-1(4H)-yl)methyl)-4-fluoro-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 26)

[0267] Compound 26 was synthesized using intermediate 22-10, 4-(bromomethyl)-3-methoxybenzonitrile, and methyl 3,5-difluoro-4-nitrobenzoate as starting materials, following the synthetic route of compound 22. Compound 26 was a white solid, 43.73 mg in total, with a yield of 67.4%. 1H NMR (300MHz, DMSO-d6) δ13.12(s,1H),8.17(s,1H),7.97(s,1H),7.52(dd,J=9.6,6.4Hz,2H),7.4 4(d,J=6.1Hz,2H),7.31(d,J=7.8Hz,1H),7.00(d,J=7.5Hz,1H),6.81(d,J=8.3Hz,1H),5.07(d,J= 8.6 Hz, 1H), 4.88 (dd, J = 15.5, 6.9 Hz, 1H), 4.74 (d, J = 15.1 Hz, 1H), 4.50 (dd, J = 22.7, 13.9 Hz, 3H), 4.39 (s, 3H), 3.87 (d, J = 7.8 Hz, 5H), 3.36 (s, 1H), 2.69 (s, 1H), 2.51 (s, 2H), 2.41 (d, J = 10.0 Hz, 1H). HRMS (ESI): calculated value C 30 H 28 FN7O4Se,[M+H] + 650.1425, found 650.1432. HPLC R =4.13min, purity 99.00%.

[0268] Example 27 Preparation of Compound 27

[0269] (S)-4-Fluoro-2-((4-(6-((2-fluoro-4-(trifluoromethyl)benzyl)seleno)pyridin-2-yl)-5,6-dihydro-1,2,4-triazin-1(4H)-yl)methyl)-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 27)

[0270] Compound 27 was synthesized using intermediate 22-10, 1-(bromomethyl)-2-fluoro-4-(trifluoromethyl)benzene, and methyl 3,5-difluoro-4-nitrobenzoate as starting materials, following the synthetic route of compound 22. Compound 27 was obtained as a white solid, yielding 37.46 mg and a yield of 59.8%.

[0271] 1H NMR (300MHz, DMSO-d6) δ13.14(s,1H), δ8.16(s,1H),7.97(s,1H),7.63(d,J=9.0Hz,2H),7.56(d, J=8.6Hz,1H),7.50(dd,J=7.4,2.8Hz,2H),7.03(d,J=7.5Hz,1H),6.84(d,J=8.3Hz,1H),5.06(d,J =7.6 Hz, 1H), 4.86 (dd, J = 15.4, 7.0 Hz, 1H), 4.73 (d, J = 14.9 Hz, 1H), 4.58–4.35 (m, 5H), 4.39–4.28 (m, 1H), 3.86 (t, J = 4.9 Hz, 2H), 3.16 (q, J = 5.7 Hz, 2H), 2.67 (d, J = 8.8 Hz, 1H), 2.39 (t, J = 8.6 Hz, 1H). HRMS (ESI): calculated value C 29 H 25 F5N6O3Se,[M+H] + 681.1146, found 681.1154. HPLC R =4.51min, purity 98.96%.

[0272] Example 28 Preparation of Compound 28

[0273] (S)-2-((4-(6-((4-chloro-2-fluorobenzyl)seleno)pyridin-2-yl)-5,6-dihydro-1,2,4-triazin-1(4H)-yl)methyl)-4-fluoro-1-(oxetan-2-ylmethyl)-1H-benzo[d]imidazole-6-carboxylic acid (Compound 28)

[0274] Compound 28 was synthesized using intermediate 22-10, 1-(bromomethyl)-4-chloro-2-fluorobenzene, and methyl 3,5-difluoro-4-nitrobenzoate as starting materials, following the synthetic route of compound 22. Compound 28 was a white solid, 39.03 mg in total, with a yield of 56.6%.

[0275] 1H NMR (300MHz, DMSO-d6) δ13.11(s,1H),8.17(s,1H),7.97(s,1H),7.55(d,J=6.8Hz,2H),7. 52–7.33(m,2H),7.19(d,J=8.3Hz,1H),7.02(d,J=7.6Hz,1H),6.83(d,J=8.3Hz,1H),5.11– 5.02 (m, 1H), 4.87 (dd, J = 15.2, 7.0 Hz, 1H), 4.73 (d, J = 15.0 Hz, 1H), 4.58–4.30 (m, 2H), 4.37 (s, 4H), 3.86 (s, 2H), 3.16 (d, J = 6.2 Hz, 2H), 2.70 (d, J = 11.6 Hz, 1H), 2.39 (t, J = 8.6 Hz, 1H). HRMS (ESI): calculated value C 28 H 25 ClF2N6O3Se,[M+H] + 647.0883, found 647.0883. HPLC R =4.83min, purity 97.94%.

[0276] Example 29 Evaluation of GLP-1R-cAMP agonistic effect

[0277] The present invention uses a cAMP signal agonist effect detection method to characterize the agonist activity of a compound on GLP-1R.

[0278] (1) Experimental reagents and instruments (see Table 2 and Table 3)

[0279] Table 2 Experimental reagents and consumables

[0280] Table 3 Experimental instruments and devices

[0281] (2) Experimental cells

[0282] In this study, we used a HEK cell line stably expressing GLP-1R (developed by Beijing Aisiyipu Biotechnology Co., Ltd., expressing the human GLP-1R gene). The number of cells used for testing was 1000 per well. All cell lines were cultured in DMEM supplemented with 10% fetal bovine serum and 100 μg / mL Hygromycin B at 37°C and 5% CO2. The old culture medium was removed and the cells were washed once with PBS. Then, 1 mL of TrypLE was added. TMExpress solution (Beijing Aisiyipu Biotechnology Co., Ltd.), incubate at 37°C for about 2 minutes. When the cells detach from the bottom of the dish, add about 2 mL of complete medium DMEM preheated at 37°C. Gently blow the cell suspension with a pipette to separate the aggregated cells. Transfer the cell suspension to a sterile centrifuge tube and centrifuge at 1000 rpm for 5 minutes to collect the cells for experiments or subculture. To maintain the physiological activity of the cells, the experimental cell fusion is controlled at about 80%. Cell passaging, recovery and freezing are carried out according to conventional methods. All operations follow the standard operating procedures for cell culture of Beijing Aisiyipu Biotechnology Co., Ltd.

[0283] (3) Experimental methods

[0284] The test used the LANCE Ultra cAMP kit.

[0285] The kit contains: Stimulation Buffer, Eu-cAMP, Detection buffer, and ULight-anti-cAMP.

[0286] Step 1: Equilibrate all reagents in the kit to room temperature before use. Prepare 1× Stimulation Buffer according to the kit instructions.

[0287] Step 2: Prepare a serial dilution of the test compound with DMSO, followed by a 10x dilution in 1x Stimulation Buffer. Set the starting concentration to 5000 nM and perform a 5-fold serial dilution to 10 groups, with each test group containing two replicate wells. Each test group served as a negative control with 0.1% DMSO and a positive control with 100 nM GLP-1 (7-37) (provided by Beijing Aisiyipu Biotechnology Co., Ltd.).

[0288] Step 3: Stably transfected cells were cultured to 80% confluency; cells were collected by trypsin digestion, counted, and inoculated into 384-well plates at 9 μL / well.

[0289] Step 4: Take 1 μL of the 10× compound diluted in step 2 and add it to the corresponding experimental wells. After centrifugation, incubate at 37°C for 30 minutes.

[0290] Step 5: Dilute Eu-cAMP to the working concentration with Detection buffer and add 5 μL / well to the corresponding experimental wells.

[0291] Step 6: Dilute ULight-anti-cAMP to the working concentration with detection buffer, then add 5 μL / well to the corresponding experimental wells; centrifuge and incubate at room temperature for 1 hour.

[0292] Step 7: After incubation, use a Biotek multifunctional microplate reader to detect the readings at 665nm and 620nm. Calculate the ratio of the readings at the corresponding wavelengths, R. 665 / 620 , calculate the average ratio (R Group ), positive reference mean ratio and negative reference mean ratio The excitation rate A was calculated according to the formula, and the excitation effect of the compound at the 5000nM level was determined. The compound with excitation effect (A 5000nM The A of the compound test concentration with an agonistic effect greater than 80% was selected and plotted against the corresponding concentration. The curve fitting and the half-maximal agonist activity concentration (EC) were calculated using the nonlinear regression method of GraphPad Prism software. 50 )calculate.

[0293] (4) Calculation method

[0294] The calculation result is rounded to 4 significant digits.

[0295] (5) Experimental results (see Table 4 and Table 5)

[0296] Reference drug Danuglipron (PF-06882961): synthesized with reference to patent WO2018109607A1.

[0297] Table 4 cAMP agonist effects of 5000 nM compounds

[0298] Table 5 EC of compounds for agonism of cAMP signal 50

[0299] The test results show that the compound of the present invention can activate the downstream signal transduction of GLP-1R by stimulating the cAMP signaling pathway, and has a complete GLP-1R agonist effect and a low half-maximal agonist concentration.

[0300] Example 30 Rat Liver Microsome Stability Test

[0301] The present invention uses an LC-MS / MS method to detect the rat liver microsome stability of compound 1, compound 17, compound 26, and PF-06882961.

[0302] (1)LC-MS / MS conditions

[0303] LC-MS / MS model: Shimadzu LCMS-8050;

[0304] Chromatographic column: GL Science ODS-3, 3 μm, 2.1 mm x 50 mm;

[0305] Mobile phase: binary high-pressure gradient elution, 0-2 min, pump A: methanol, 20%; pump B: water (containing 0.1% formic acid), 80%; 2-5 min: pump A: methanol, 80%; pump B: water (containing 0.1% formic acid), 20%; 5-7 min, pump A: methanol, 20%; pump B: water (containing 0.1% formic acid), 80%;

[0306] Detection wavelength: 254 nm; detection time: 7 min; injection volume: 5 μL;

[0307] Calculation method: clarithromycin-internal standard method;

[0308] Internal standard concentration: 10 ng / mL;

[0309] (2) Establishment of standard curve

[0310] Prepare a 100 mM MgCl₂ solution in PBS, a 5 mM NADPH solution in PBS, a 5 mg / mL rat liver microsome (rHLM, purchased from Shanghai Quanyang Biotechnology Co., Ltd.) test solution diluted in PBS (stored at 4°C), a 5 mg / mL inactivated rat liver microsome (rHLM, inactivated by heating at 60°C for 1 hour) test solution diluted in PBS (stored at 4°C), and a 1 mM stock solution of the compound in MeOH. Dilute the stock solution of the compound to an 80 μM test solution using PBS, and then perform a two-fold serial dilution in PBS to ten groups.

[0311] Test system: 120 μL PBS, 20 μL 5 mg / mL inactivated liver microsomes, 10 μL 100 mM MgCl₂ solution, 10 μL compound, mix well, incubate at 37°C for 5 min, and add 40 μL NADPH solution. A total of 200 μL was prepared. The test system consisted of a 10-fold dilution of rHLM, a 20-fold dilution of MgCl₂, a 20-fold dilution of compound, and a 5-fold dilution of NADPH.

[0312] Take a 10 μL sample and add 30 μL of a 12 ng / mL clarithromycin diluted in acetonitrile as an internal standard. Vortex to mix thoroughly. The final concentration of the compound to be tested is an 80-fold dilution of the stock solution. Precipitate the protein, centrifuge at 12,000 rpm at 4°C for 5 minutes, and collect 20 μL of the supernatant. The final concentration to be tested should be between 1000 ng / mL and 0.1 ng / mL. Inject 5 μL of the sample for LC-MS / MS analysis. Plot a standard curve using the internal standard method by plotting concentration ratio (C) against area ratio (A).

[0313] The standard curve formulas for compound 1 and PF-06882961 were prepared using the above test method. The fitting results showed that the peak area response of compound 1 and PF-06882961 was linearly related to the concentration in the range of 0.1 ng / mL-1000 ng / mL, and the fitting regression degree was good.

[0314] The standard curve of compound 1 is shown in FIG1 , and the standard curve of PF-06882961 is shown in FIG2 .

[0315] (3) Test method

[0316] The test method described in step (2) was used for detection in three parallel groups. The concentration of the test sample was 4 μg / mL. After the NADPH solution was added to initiate the reaction, the timing was started and samples were taken at 0, 5, 10, 15, 20, 30, 45, 60, 90, and 120 min. The residual concentration of the compound at each time point was calculated, and the compound retention percentage (HLMRemaining, %) was calculated with the compound concentration at 0 min as 100%. The average value of the natural logarithm of the retention percentage was linearly fitted with the incubation time to obtain the elimination rate constant k (min -1 ) and SEM. Calculate the clearance-related parameters according to the following formula:

[0317] Half-life t 1 / 2 (min) = 0.693 / |k|; intrinsic clearance CL int (mL / min / kg)=(0.693 / t 1 / 2 )*(1 / C rHLM )*scaling factor (rat: 1792);

[0318] The degradation of the compound under rHLM conditions 1 / 2 and CL int The stability of the compound in rat liver microsomes was characterized. The results are shown in Table 6:

[0319] Table 6 rHLM stability of compounds

[0320] The test results showed that the stability of compound 1, compound 17 and compound 26 in rat liver microsomes was better than that of the positive molecule PF-06882961 in rat liver microsomes.

[0321] Example 31 Pharmacokinetics Test

[0322] The present invention uses the LC-MS / MS method to calculate the resident concentrations (c) of compound 1 and PF-06882961 in adult rat plasma at different time points (t) under different administration conditions, draws ct curves for corresponding conditions / corresponding compounds, and uses Winnolin 7.0 software non-modal model to fit and analyze the ct parameters.

[0323] (1) Preliminary preparation

[0324] Rats: SD strain, male, 6-8 weeks old, weighing 180 g-220 g;

[0325] Administration: intravenous injection (1 mg / kg or 10 mg / kg); oral administration (5 mg / kg or 50 mg / kg);

[0326] Prepare solvent formula: 10% DMSO + 40% PEG-400 + 50% normal saline (containing 20% ​​w / v hydroxypropyl-β-cyclodextrin), solution concentration is 1 mg / mL or 5 mg / mL;

[0327] (2)LC-MS / MS conditions

[0328] LC-MS / MS model: Shimadzu LCMS-8050;

[0329] Chromatographic column: GL Science ODS-3, 3 μm, 2.1 mm x 50 mm;

[0330] Mobile phase: binary high-pressure gradient elution, 0-2 min, pump A: methanol, 20%; pump B: water (containing 0.1% formic acid), 80%; 2-5 min: pump A: methanol, 80%; pump B: water (containing 0.1% formic acid), 20%; 5-7 min, pump A: methanol, 20%; pump B: water (containing 0.1% formic acid), 80%;

[0331] Detection wavelength: 254 nm; detection time: 7 min; injection volume: 10 μL or 1 μL;

[0332] Calculation method: clarithromycin-internal standard method;

[0333] Internal standard concentration: 10 ng / mL;

[0334] (3) Establishment of standard curve

[0335] Test compound standard concentrations of 10,000 ng / mL, 1,000 ng / mL, 500 ng / mL, 200 ng / mL, 100 ng / mL, 50 ng / mL, 10 ng / mL, 1 ng / mL, and 0.1 ng / mL were prepared in acetonitrile. A 20 μL sample was diluted with 40 μL of a 20 ng / mL acetonitrile solution and 20 μL of blank rat plasma. Analyzed by LC-MS as described previously, 10 μL of sample was injected each time, and the corresponding ion peak areas were calculated. Using the internal standard method, a standard curve was constructed for the corresponding compound, plotting concentration ratio (C) against area ratio (A).

[0336] (4) Sampling and processing calculation

[0337] Rats were weighed and the dosing volume was calculated. Blood was collected at 1 minute, 5 minutes, 10 minutes, 30 minutes, 45 minutes, 60 minutes, 120 minutes, 240 minutes, 360 minutes, 480 minutes, 540 minutes, and 1440 minutes, for a total of 12 time points. Four parallel experimental groups were set up for each dosing group. The timer started at the end of the dosing time point. At the set blood collection time point, 100 μL to 200 μL of blood was collected from the rat's orbital venous plexus. Plasma was separated according to the universal plasma separation method (4500 rpm, RT, 5 minutes). Then 20 μL of sample plasma was drawn. 40 μL of 20 ng / mL clarithromycin acetonitrile solution and 20 μL of acetonitrile were added to the sample plasma and vortexed to mix and precipitate the protein. Centrifuge at 12000 rpm and 4°C for 5 min, aspirate 40 μL of the supernatant, and analyze according to the above LC-MS method. Inject 10 μL of each sample, calculate the corresponding ion peak area, and use the internal standard method standard curve to calculate the compound concentration.

[0338] The CT curves of compound concentration versus time were plotted, and the pharmacokinetic parameters were calculated using the non-compartmental statistical moment method using Winnolin 7.0 software. Wherein the subscript iv represents intravenous injection, and po represents oral administration. max and T max All are measured values, AUC last The value is calculated by the trapezoidal area method, the area under the curve AUC for 0-+∞ time int_obs =AUC last +C last / HL_Lambda_z, where C last represents the final identifiable minimum effective concentration, and HL_Lambda_z represents the elimination phase half-life. Oral bioavailability F = (AUC int_obsp.o. / D p.o. ) / (AUC int_obsi.v. / D i.v.)*100%, where AUC represents the area under the curve, and D represents the administered dose.

[0339] (5) Test parameters

[0340] The CT curve of PF-06882961 is shown in Figure 3, the CT curve of the low-dose group of Compound 1 is shown in Figure 4, and the CT curve of the high-dose group of Compound 1 is shown in Figure 5. The PK parameters obtained from the test are shown in Table 7.

[0341] Table 7 Pharmacokinetic parameters of the compounds

[0342] *PF-06882961 is administered orally as its Tris salt, synthesized using the method disclosed in patent WO2018109607A1;

[0343] **Oral group V z_obs and CL obs The data show apparent parameters.

[0344] Data showed that the low-dose group of compound 1 had a short intravenous half-life and a long oral half-life. Under intravenous administration, the drug clearance rate was rapid, and the plasma drug inventory was lower than that of the active drug. Under oral administration, drug absorption and metabolism were balanced, and the plasma drug inventory was significantly increased compared to intravenous administration, indicating tissue drug accumulation and sustained release. This resulted in an oral bioavailability that was superior to the active drug PF-06882961, at 20.84%. The high-dose group had moderate intravenous and oral half-lives of 1.37 hours and 2.65 hours, respectively. The in vivo absorption and average AUC values ​​were both higher than those of the active drug, and the oral bioavailability of the high-dose group was higher than that of PF-06882961.

[0345] The above experimental results evaluated the in vitro drug-forming properties of compound 1.

[0346] Example 32 Pharmacokinetics Test

[0347] The present invention uses the LC-MS / MS method to calculate the resident concentration (c) of compound 17 in adult rat plasma at different time points (t) under different administration conditions, draws the ct curve of the corresponding conditions / corresponding compounds, and uses the Winnolin7.0 software non-mode model to fit and analyze the ct parameters.

[0348] (1) Preliminary preparation

[0349] Rats: SD strain, male, 6-8 weeks old, weighing 180 g-220 g;

[0350] Administration: intravenous injection (5 mg / kg); oral administration (30 mg / kg);

[0351] Prepare solvent formula: 10% DMSO + 40% PEG-400 + 50% saline (containing 20% ​​w / v hydroxypropyl-β-cyclodextrin), solution concentration is 5 mg / mL;

[0352] (2)LC-MS / MS conditions

[0353] Same as Example 31.

[0354] (3) Establishment of standard curve

[0355] Same as Example 31.

[0356] (4) Sampling and processing calculation

[0357] Same as Example 31.

[0358] (5) Test parameters

[0359] The PK parameters obtained from the test are shown in Table 8.

[0360] Table 8 Pharmacokinetic parameters of the compounds

[0361] The pharmacokinetic properties of compound 17 were evaluated using intravenous injection at 5 mg / kg and oral gavage at 30 mg / kg. The results showed that compound 17 had moderate intravenous and oral half-lives, with relatively maintained mean plasma drug exposure, apparent volume of distribution, and clearance. The oral bioavailability reached 28.91%. These experimental results demonstrate the in vitro druggability of compound 17.

[0362] Thus, we have obtained a small molecule GLP-1R agonist with good GLP-1R agonist activity and acceptable oral administration efficacy, expanding the application of selenium in drug design and synthesis. However, the structural design and modification of the selenium linker compound series represented by Compounds 1 and 17 remains relatively conservative, and there is still potential for further improvement in druggable properties such as bioavailability and cardiotoxicity.

[0363] As described above, although the present invention has been shown and described with reference to specific preferred embodiments, it should not be construed as limiting the present invention itself. Various changes may be made to it in form and detail without departing from the spirit and scope of the present invention as defined in the appended claims.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt, enantiomer, stereoisomer, hydrate, solvate or polymorph thereof: in, Ring A is selected from C 3-8 Cycloalkyl, C 2-9 Heterocyclic group, C 5-12 Spirobicyclic, C 5-12 Spiroheterobicyclic, C 5-12 Fused bicyclic group, C 5-12 Fused heterobicyclic group, C 5-12 Bridged ring or C 5-12 bridged heterocyclic group; L is selected from -C(O)-, -O-, -S-, -CR 4 R 5 or -NR 6 -; R 4 , R 5 are each independently selected from hydrogen, deuterium, halogen, cyano, C1-C6 alkyl, C1-C6 alkoxy, -NR 5a R 5b , 6-10 membered aryl, 5-8 membered heteroaryl, 3-8 membered saturated or partially saturated cycloalkyl, 3-8 membered saturated or partially saturated heterocyclic group; the C1-C6 alkyl or C1-C6 alkoxy is optionally substituted by one or more of the following substituents: halogen, oxo, cyano, hydroxyl, C3-C6 saturated or partially saturated cycloalkyl; the aryl, heteroaryl, saturated or partially saturated cycloalkyl, saturated or partially saturated heterocyclic group is optionally substituted by one or more of the following substituents: halogen, oxo, cyano or -NR 5a R 5b ; R 5a , R 5b Each is independently selected from hydrogen or C1-C6 alkyl; R 6 Selected from hydrogen, C1-C6 alkyl, C3-C10 cycloalkyl, heterocyclic group, -S(O)2R 6a or -S(O)2N(R 6a )(NR 6b ); the cycloalkyl or heterocyclic group is optionally substituted by C1-C6 alkyl, -F or cyano; R 6a , R 6b Each is independently selected from -H, C1-C6 alkyl or C3-C10 cycloalkyl; R 1 C1-C6 alkyl substituted with 3-6 membered heterocyclyl, C1-C6 alkyl substituted with C3-C6 cycloalkyl, C1-C6 alkyl substituted with 5-8 membered heteroaryl, C1-C6 alkyl substituted with C1-C6 alkoxy, 3-6 membered heterocyclyl, C3-C6 cycloalkyl or 5-8 membered heteroaryl; the 3-6 membered heterocyclyl, C3-C6 cycloalkyl, 5-8 membered heteroaryl or C1-C6 alkoxy is optionally replaced by one of the following: substituted by one or more substituents: H, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 alkylamino, cyano, cyano-substituted C1-C6 alkyl, hydroxy, hydroxy-substituted C1-C6 alkyl, halo-substituted C1-C6 alkyl, halo-substituted C3-C6 cycloalkyl, halo-substituted C1-C6 alkoxy, halo-substituted C1-C6 alkylamino, acyl, amide, aminoacyl, sulfonyl or amino; R 2 Each is independently selected from a hydrogen atom, deuterium, halogen, cyano, hydroxyl, oxo, C1-C6 alkyl, C1-C6 alkoxy, amino, 6-10 membered aryl, 5-8 membered heteroaryl, 3-8 membered saturated or partially saturated cycloalkyl, 3-8 membered saturated or partially saturated heterocyclic group; R 3 Each is independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxyl, cycloalkyl, heterocyclyl, aryl or heteroaryl; the alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, cyano, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl; X 1 , X 2 , X 3 and X 4 The same or different, and each independently selected from -CR 7 or N atoms; R 7 is selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclic, aryl or heteroaryl; G is selected from G1 or G2; the structural formulas of G1 and G2 are as follows: Ring B and Ring C are each independently selected from a 6-12 membered aryl group or a 6-12 membered heteroaryl group; Y and Z are each independently selected from -CH2-, -S-, -SO-, -SO2-, -(O=S=NH)-, -Se- or -Se(O)-; R 8 , R 11 are the same or different and are each independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl; the alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl being optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, cyano, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl; R 9 , R 10 , R 12 , R 13 are the same or different and are each independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl; EE is selected from -COOH or a carboxyl surrogate; the carboxyl surrogate is selected from: m is 0, 1, 2 or 3; n is 0, 1, 2, 3 or 4; o is 0, 1, 2, 3, 4, or 5; p is 0, 1, 2, 3, 4 or 5.

2. The compound according to claim 1, characterized in that Ring A is selected from C 3-8 Cycloalkyl or C 2-9 Heterocyclic group; L is selected from -CR 4 R 5 ; R 1 Selected from C1-C6 alkyl substituted by 3-6 membered heterocyclyl, C1-C6 alkyl substituted by C3-C6 cycloalkyl, C1-C6 alkyl substituted by 5-8 membered heteroaryl, C1-C6 alkyl substituted by C1-C6 alkoxy; EE is selected from -COOH or a carboxyl surrogate.

3. The compound according to claim 2, characterized in that Ring A is selected from C 2-9 Heterocyclic group; Y and Z are each independently selected from -Se- or -Se(O)-; EE is selected from -COOH, 4. The compound according to claim 1, characterized in that Selected from:

5. A pharmaceutical composition, characterized in that: The invention comprises the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt, enantiomer, stereoisomer, hydrate, solvate or polymorph thereof and a pharmaceutically acceptable carrier or excipient.

6. Use of the compound according to any one of claims 1 to 4 in the preparation of a GLP-1 receptor agonist, or in the preparation of a medicament for preventing and / or treating diseases associated with GLP-1 receptor stimulation.

7. The use according to claim 6, characterized in that The diseases associated with stimulating GLP-1 receptors include diabetes, diabetic complications, metabolic syndrome, obesity, non-alcoholic fatty liver disease, non-alcoholic fatty hepatitis, Parkinson's disease, Alzheimer's disease, hypertension, hyperlipidemia, atherosclerosis, cardiovascular risk, coronary heart disease or stroke.

8. A compound of formula II or a pharmaceutically acceptable salt, enantiomer, stereoisomer, hydrate, solvate or polymorph thereof: in, Ring A is selected from C 3-8 Cycloalkyl, C 2-9 Heterocyclic group, C 5-12 Spirobicyclic, C 5-12 Spiroheterobicyclic group, C 5-12 Fused bicyclic group, C 5-12 Fused heterobicyclic group, C 5-12 Bridged ring or C 5-12 bridged heterocyclic group; L is selected from -C(O)-, -O-, -S-, -CR 4 R 5 or -NR 6 -; R 4 , R 5 are each independently selected from hydrogen, deuterium, halogen, cyano, C1-C6 alkyl, C1-C6 alkoxy, -NR 5a R 5b , 6-10 membered aryl, 5-8 membered heteroaryl, 3-8 membered saturated or partially saturated cycloalkyl, 3-8 membered saturated or partially saturated heterocyclic group; the C1-C6 alkyl or C1-C6 alkoxy is optionally substituted by one or more of the following substituents: halogen, oxo, cyano, hydroxyl, C3-C6 saturated or partially saturated cycloalkyl; the aryl, heteroaryl, saturated or partially saturated cycloalkyl, saturated or partially saturated heterocyclic group is optionally substituted by one or more of the following substituents: halogen, oxo, cyano or -NR 5a R 5b ; R 5a , R 5b Each is independently selected from hydrogen or C1-C6 alkyl; R 6 Selected from hydrogen, C1-C6 alkyl, C3-C10 cycloalkyl, heterocyclic group, -S(O)2R 6a or -S(O)2N(R 6a )(NR 6b ); the cycloalkyl or heterocyclic group is optionally substituted by C1-C6 alkyl, -F or cyano; R 6a , R 6b Each is independently selected from -H, C1-C6 alkyl or C3-C10 cycloalkyl; R 1 C1-C6 alkyl substituted with 3-6 membered heterocyclyl, C1-C6 alkyl substituted with C3-C6 cycloalkyl, C1-C6 alkyl substituted with 5-8 membered heteroaryl, C1-C6 alkyl substituted with C1-C6 alkoxy, 3-6 membered heterocyclyl, C3-C6 cycloalkyl or 5-8 membered heteroaryl; the 3-6 membered heterocyclyl, C3-C6 cycloalkyl, 5-8 membered heteroaryl or C1-C6 alkoxy is optionally replaced by one of the following: substituted by one or more substituents: H, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 alkylamino, cyano, cyano-substituted C1-C6 alkyl, hydroxy, hydroxy-substituted C1-C6 alkyl, halo-substituted C1-C6 alkyl, halo-substituted C3-C6 cycloalkyl, halo-substituted C1-C6 alkoxy, halo-substituted C1-C6 alkylamino, acyl, amide, aminoacyl, sulfonyl or amino; R 2 Each is independently selected from hydrogen, deuterium, halogen, cyano, hydroxyl, oxo, C1-C6 alkyl, C1-C6 alkoxy, amino, 6-10 5- to 8-membered aryl, 5- to 8-membered heteroaryl, 3- to 8-membered saturated or partially saturated cycloalkyl, 3- to 8-membered saturated or partially saturated heterocyclic group; R 3 Each is independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxyl, cycloalkyl, heterocyclyl, aryl or heteroaryl; the alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, cyano, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl; X 1 , X 2 , X 3 and X 4 The same or different, and each independently selected from -CR 7 or N atoms; R 7 is selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclic, aryl or heteroaryl; G is selected from G1 or G2; the structural formulas of G1 and G2 are as follows: Ring B and Ring C are each independently selected from a 6-12 membered aryl group or a 6-12 membered heteroaryl group; Y and Z are each independently selected from -CH2-, -S-, -SO-, -SO2-, -(O=S=NH)-, -Se- or -Se(O)-; R 8 , R 11 are the same or different and are each independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl; the alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl being optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, cyano, amino, nitro, cyano, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl; R 9 , R 10 , R 12 , R 13 are the same or different and are each independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl; R 14 each independently selected from hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted with one or more of the following substituents: halogen, alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cyano, amino, nitro, hydroxy, cycloalkyl, heterocyclyl, aryl or heteroaryl; EE is selected from -COOH or a carboxyl surrogate; the carboxyl surrogate is selected from: m is 0, 1, 2 or 3; n is 0, 1, 2, 3 or 4; o is 0, 1, 2, 3, 4, or 5; p is 0, 1, 2, 3, 4 or 5; q is 0, 1, 2, or 3.

9. The compound according to claim 8, characterized in that Selected from:

10. Use of the compound according to claim 8 or 9 in the preparation of a GLP-1 receptor agonist, or in the preparation of a medicament for preventing and / or treating diseases associated with GLP-1 receptor stimulation.

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