Inhibitors of isr and methods of making and using the same

Abstract

The application discloses an ISR inhibitor and a preparation method and application thereof, and belongs to the technical field of biological medicines. The compound of the application is a compound with the structure of formula (I) or formula (II), a stereoisomer thereof, a geometric isomer thereof, a tautomer thereof, a pharmaceutically acceptable salt thereof, a prodrug thereof, a hydrate thereof or a solvate thereof. The compound provided in the application can be effectively combined with eIF2B, has good ATF4 inhibiting capacity, and can better restore protein synthesis when ISR is stimulated. Therefore, the compound of the application can be used as an ISR inhibitor and for treating various related diseases.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a class of compounds with ISR inhibitory activity, their preparation methods, and applications. Background Technology

[0002] The integrated stress response (ISR) is a signaling network induced by protein homeostasis disruption. It primarily controls the rate of protein synthesis, helping cells, tissues, and organisms adapt to changing environments and maintain health. The biological function of ISR is to restore intracellular homeostasis under stress; the accumulation of misfolded proteins activates the ISR response. This pathway operates at the translation initiation level, inhibiting overall protein synthesis while simultaneously promoting the translation of several proteins involved in cellular recovery.

[0003] Activation of the interstitial reticular formation (ISR) is a common factor in many neurodegenerative diseases. ISR activation has been detected in a wide range of brain diseases, including Alzheimer's disease, Parkinson's disease, autistic spectrum disorder (ASD), amyotrophic lateral sclerosis (ALS), and traumatic brain injury.

[0004] ISR also plays a role in other diseases. Mutations in ADAR1 can cause immune diseases (including Aicardi-Goutières syndrome), and pharmacological inhibition of ISR increased the median survival rate of ADAR-mutant mice by 3-fold and restored them to normal body weight. Another study showed that targeted inhibition of ISR can regulate the M1 / M2 phenotype of microglia, exerting a novel anti-inflammatory effect on microglia and representing a potential therapeutic target for neuroinflammation following surgical brain injury. Inhibition of ISR can also alleviate pulmonary fibrosis and treat spinal cord injuries and ischemic diseases (such as myocardial infarction and stroke).

[0005] ISR signaling regulates the concentration of the intracellular eIF2 ternary complex (TC). TC is a heterotrimer composed of the eukaryotic translation initiation factor eIF2, GTP, and charged methionyl-tRNAi (Met-tRNAi). Translation initiation is a multi-step process. The mRNA-protein complex (mRNP) recruits a multifactor complex (MFC) containing initiation factors eIF1, eIF3, and eIF5, as well as the eIF2 complex binding GTP and initiation tRNA, promoting the transfer of the ternary complex to the 40S ribosomal subunit. The MFC is most commonly recruited to the mRNA by the eIF4F complex, which recognizes and binds to the 50cap structure and localizes the MFC to scan from the 50 end to the AUG start codon. Once AUG is recognized, the hydrolysis of GTP by eIF2 allows the 40S ribosome to enter the translation initiation phase, leading to the recruitment of the 60S subunit and the elongation phase of translation initiation. After eIF2–GDP is released from the ribosome, GDP is exchanged for GTP by the guanine nucleotide-exchange factor (GEF) eIF2B, preparing the eIF2 complex for the next initiation. The activity of eIF2B is regulated by phosphorylation of its substrate eIF2, binding of nucleotides and cofactors to eIF2B, and phosphorylation of eIF2B itself. eIF2B activators can promote the formation of eIF2B decameric protein, reverse the effects of ISR, drive translation initiation, and restore protein synthesis.

[0006] Therefore, research on novel compounds with ISR inhibitory activity is of great significance for the treatment of various diseases. Summary of the Invention

[0007] The purpose of this invention is to provide a class of compounds with ISR inhibitory activity, their preparation methods, and applications.

[0008] To achieve the above objectives, the present invention adopts the following technical solution:

[0009] This invention provides compounds having the structure of formula (I) or formula (II), their stereoisomers, their geometric isomers, their tautomers, their pharmaceutically acceptable salts, their prodrugs, their hydrates, or their solvates.

[0010]

[0011] in:

[0012] A is a halogen-substituted biaromatic heterocycle;

[0013] D is selected from

[0014] L is

[0015] W is a five-membered heterocyclic ring;

[0016] n is 0, 1, or 2;

[0017] m is 0 or 1;

[0018] B is a halogen-substituted benzene ring. K is selected from

[0019] G is selected from In some embodiments,

[0020] A is selected from

[0021] W selected B is selected from

[0022] In some specific embodiments, the present invention provides, but is not limited to, the following compounds: stereoisomers, geometric isomers, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, or solvates thereof:

[0023]

[0024]

[0025]

[0026] In this invention, the pharmaceutically acceptable salts include acid addition salts formed by compounds having the structure of formula (I) or formula (II) with the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, trifluoroacetic acid, lactic acid, oxalic acid, adipic acid, glutaric acid, malonic acid, maleic acid, succinic acid, fumaric acid, tartaric acid, citric acid, palmitic acid, benzoic acid, methanesulfonic acid, p-toluenesulfonic acid, salicylic acid, phenylacetic acid, and mandelic acid, and also include acid salts of inorganic bases.

[0027] The present invention also provides a method for preparing the compound shown in formula (I) or formula (II):

[0028]

[0029] Method 1: Compound (III) and compound (V) undergo a condensation reaction under the action of a condensing agent to prepare compound (I);

[0030] Method 2: In a polar aprotic solvent, under the action of cesium carbonate, compound (III) and compound (V) undergo a substitution reaction to prepare compound (I);

[0031] Method 3: Under the action of a condensing agent, compounds (VI) and (VII) undergo a condensation reaction to prepare compound (II);

[0032] The definitions of A, B, D, L, W, K, G, m, and n are as described above.

[0033] The present invention also provides a pharmaceutical composition comprising the active ingredient or main active ingredient of the compound described herein, its stereoisomer, its geometric isomer, its tautomer, its pharmaceutically acceptable salt, its prodrug, its hydrate or its solvate, and supplemented with a pharmaceutically acceptable carrier.

[0034] The present invention also provides the use of the compounds described herein, their stereoisomers, their geometric isomers, their tautomers, their pharmaceutically acceptable salts, their prodrugs, their hydrates or solvates thereof in the preparation of medicaments for the treatment and / or prevention of ISR-mediated diseases.

[0035] The diseases mentioned are neurodegenerative diseases, autoimmune diseases, musculoskeletal diseases, inflammatory diseases, and genetic disorders. Preferably, the neurodegenerative diseases are selected from: cerebral ischemia, brain injury, epilepsy, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, different types of spinocerebellar ataxia, autism spectrum disorders, white matter ablation disorders, intellectual disability syndromes, prion diseases, Creutzfeldt-Jakob disease, stroke, chronic traumatic encephalopathy, spinal cord injury, dementia, frontotemporal dementia (FTD), depression, or social behavioral disorders; preferably, the autoimmune diseases are selected from: systemic lupus erythematosus, type 1 diabetes, multiple sclerosis, or rheumatoid arthritis; preferably, the musculoskeletal diseases are selected from: Myopathy, muscular dystrophy, muscle atrophy, muscle wasting, or sarcopenia; preferably, the inflammatory disease is selected from: arthritis, psoriatic arthritis, psoriasis, juvenile idiopathic arthritis, asthma, allergic asthma, bronchial asthma, tuberculosis, chronic bronchial disease, cystic fibrosis, glomerulonephritis, membranous nephropathy, sarcoidosis, vasculitis, ichthyosis, graft rejection, interstitial cystitis, atopic dermatitis, or inflammatory bowel disease; preferably, the genetic condition is selected from: Down syndrome or MEHMO syndrome (intellectual disability, seizures, hypogonadism, microcephaly, and obesity).

[0036] The compounds provided by this invention can effectively bind to eIF2B, exhibit good ATF4 inhibitory activity, and can effectively restore protein synthesis upon ISR stimulation. Therefore, the compounds of this invention can be used as ISR inhibitors to treat various related conditions. This invention provides a new option for clinically screening and / or preparing drugs for the treatment and / or prevention of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, aging, autism spectrum disorder, white matter ablation disorders, dementia, and frontotemporal dementia (FTD). Detailed Implementation

[0037] The present invention will be further described in detail below with reference to specific embodiments, but this should not be construed as limiting the present invention. Any modifications or substitutions made to the methods, steps, or conditions of the present invention without departing from the spirit and substance of the invention are within the scope of the present invention. Experimental methods and reagents not specified in the embodiments are performed according to conventional conditions in the art.

[0038] Example 1

[0039] Synthesis of the intermediate trans-(4-(5-chlorobenzofuran-2-carbamate)cyclohexyl)tert-butyl carbamate

[0040]

[0041] Step 1: Synthesis of ethyl 5-chlorobenzofuran-2-carboxylate

[0042] 5-Chlorosalicylic acid (0.2 g, 1.28 mmol) was dissolved in N,N-dimethylformamide (10 mL), followed by the addition of ethyl bromoacetate (0.32 g, 1.92 mmol) and potassium carbonate (0.41 g, 3.19 mmol). The mixture was heated to 120 °C and reacted for 10 h. The product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 200 / 1) to give 204 mg of a white solid, yielding 71.09%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 7.89 (dd, J=2.2, 0.6Hz, 1H), 7.80 (dt, J=8.9, 0.8Hz, 1H), 7.7 3(d,J=0.9Hz,1H),7.56(dd,J=8.9,2.3Hz,1H),4.37(q,J=7.1Hz,2H),1.34(t,J=7.1Hz,3H).

[0043] Step 2: Synthesis of 5-chlorobenzofuran-2-carboxylic acid

[0044] Ethyl 5-chlorobenzofuran-2-carboxylate (3.15 g, 14.00 mmol) was dissolved in tetrahydrofuran / ethanol (15 mL / 10 mL), and lithium hydroxide (503 mg, 21.00 mmol) was added. The reaction was carried out at room temperature for 5 h. Under ice bath conditions, the pH was adjusted to 3 with 1 mol / L hydrochloric acid, filtered, and the filter cake was washed with water and a small amount of ethyl acetate, and dried to give 2.29 g of white solid, with a yield of 83.02%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 13.78 (s, 1H), 7.87 (d, J = 2.2Hz, 1H), 7.77 (d, J = 8.8Hz, 1H), 7.63 (d, J = 0.9Hz, 1H), 7.53 (dd, J = 8.9, 2.2Hz, 1H).

[0045] Step 3: Synthesis of trans-(4-(5-chlorobenzofuran-2-carbamate)cyclohexyl)tert-butyl carbamate

[0046] 5-Chlorobenzofuran-2-carboxylic acid (0.60 g, 3.05 mmol) was dissolved in 15 mL of N,N-dimethylformamide, followed by the sequential addition of HATU (2.32 g, 6.10 mmol), N-Boc-trans-1,4-cyclohexanediamine (0.65 mg, 3.05 mmol), and DIPEA (1.58 g, 12.21 mmol). The mixture was stirred at room temperature for 3 h. 30 mL of water was added, and the mixture was filtered. The filter cake was washed with a small amount of dichloromethane and dried to give 0.948 g of a white solid, yield 79.06%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.62 (d, J = 8.1Hz, 1H), 7.87 (d, J = 2.2Hz, 1H), 7.70 (d, J = 8.8Hz, 1H), 7.56 -7.45 (m, 2H), 6.79 (d, J = 8.0Hz, 1H), 3.79 -3.63(m,1H),3.20(d,J=10.6Hz,1H),1.81(d,J=11.5Hz,4H),1.48(d,J=12.5Hz,2H),1.38(s,9H),1.33-1.20(m,2H).

[0047] Example 2

[0048] Synthesis of the intermediate trans-N-(4-aminocyclohexyl)-5-chlorobenzofuran-2-carboxamide

[0049]

[0050] 948 mg (2.41 mmol) of trans-(4-(5-chlorobenzofuran-2-carbamate)cyclohexyl)carbamate tert-butyl ester was dissolved in 10 mL of dichloromethane, and 1 mL of trifluoroacetic acid was added. The reaction mixture was reacted at room temperature for 8 h. The reaction solution was concentrated under reduced pressure, the pH was adjusted to 10 with ammonia, filtered, the filter cake was washed, and dried to give 586 mg of a pale yellow solid, with a yield of 82.95%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.68 (d, J = 8.0Hz, 1H), 7.88 (d, J = 2.7Hz, 1H), 7.70 (d, J = 8.8Hz, 1H), 7. 55-7.45(m,2H),3.75(s,1H),2.99(d,J=8.4Hz,1H),2.01-1.85(m,4H),1.42(qd,J=11.4,5.6Hz,4H).

[0051] Example 3

[0052] Synthesis of ethyl 5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxylate

[0053]

[0054] Step 1: Synthesis of 4-chlorobenzoylhydrazine

[0055] Ethyl 4-chlorobenzoate (5.00 g, 27.08 mmol) was dissolved in methanol (50 mL), and 80% hydrazine hydrate (6.78 g, 135.41 mmol) was added. The mixture was heated to 80 °C and refluxed for 12 h. The reaction solution was concentrated, washed with a small amount of petroleum ether / ethyl acetate, filtered, and dried to give 4.10 g of a white solid, with a yield of 88.65%. 1 H NMR (400MHz, DMSO-d6) δ (ppm): 9.73 (s, 1H), 7.73-7.66 (m, 2H), 7.48-7.41 (m, 2H), 5.87 (s, 2H).

[0056] Step 2: Synthesis of ethyl 2-(2-(4-chlorobenzoyl)hydrazino)-2-oxoethyl acetate

[0057] 4-Chlorobenzoylhydrazide (1.96 g, 11.51 mmol) was dissolved in dichloromethane (15 mL), and triethylamine (3.20 mL, 23.03 mmol) was added. Then, oxaloyl chloride monoethyl ester (3.14 g, 23.03 mmol) was slowly added dropwise under ice bath conditions. After the addition was complete, the ice bath was removed, and the reaction was allowed to proceed at room temperature for 1 h. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1 to 1 / 1) to give 2.36 g of white solid, yield 75.74%.1 H NMR (300MHz, DMSO-d6) δ (ppm): 10.98 (s, 1H), 10.70 (s, 1H), 7.94-7.87 (m, 2H), 7.64-7.58 (m, 2H), 4.22 (q, J = 7.1Hz, 2H), 1.31 (t, J = 7.1Hz, 3H).

[0058] Step 3: Synthesis of ethyl 5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxylate

[0059] Ethyl 2-(2-(4-chlorobenzoyl)hydrazino)-2-oxoethyl acetate (2.11 g, 7.80 mmol) was dissolved in acetonitrile (15 mL), and phosphorus oxychloride (3.59 g, 23.39 mmol) was slowly added. The mixture was heated to 85 °C and the tube was sealed for 8 h. Water was slowly added under ice bath conditions, followed by extraction with ethyl acetate. The extract was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 80 / 1) to give 807 mg of a white solid, with a yield of 40.97%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.13-8.05 (m, 2H), 7.77-7.69 (m, 2H), 4.47 (q, J = 7.1Hz, 2H), 1.38 (t, J = 7.1Hz, 3H).

[0060] Example 4

[0061] Synthesis of trans-N-(4-(5-chlorobenzofuran-2-carbamate)cyclohexyl)-5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxamide (I-1)

[0062]

[0063] Step 1: Synthesis of 5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxylic acid

[0064] Following the method described in step 2 of Example 1, 149 mg of a white solid was obtained, with a yield of 57.91%. 1 H NMR (300MHz, Chloroform-d) δ (ppm): 8.50 (s, 1H), 8.08-8.02 (m, 2H), 7.57-7.50 (m, 2H).

[0065] Step 2: Synthesis of trans-N-(4-(5-chlorobenzofuran-2-carbamate)cyclohexyl)-5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxamide (Ⅰ-1)

[0066] 5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxylic acid (59 mg, 263 μmol) was dissolved in acetonitrile (10 mL), and oxalyl chloride (25 μL, 289 μmol) was added under ice bath conditions, followed by 1 drop of N,N-dimethylformamide. The reaction was allowed to proceed for 30 min. trans-N-(4-aminocyclohexyl)-5-chlorobenzofuran-2-carboxamide (77 mg, 263 μmol) was dissolved in dichloromethane (5 mL), and N,N-diisopropylethylamine (102 mg, 784 μmol) was added. The reaction mixture was then slowly added under ice bath conditions. After the addition was complete, the ice bath was removed, and the reaction was allowed to proceed at room temperature for 3 h. The mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane / methanol = 200 / 1) to give 75 mg of a white solid, with a yield of 57.11%. 1 H NMR(300MHz,DMSO-d6)δ(ppm):9.31(d,J=8.2Hz,1H),8.68(d,J=8.1Hz,1H), 8.13(d,J=2.0Hz,1H),8.10(d,J=2.0Hz,1H),7.88(d,J=2.2Hz,1H),7.76-7.6 8(m,3H),7.54(d,J=1.0Hz,1H),7.49(dd,J=8.8,2.2Hz,1H),3.79(d,J=9.1Hz,2H),1.91(d,J=9.4Hz,4H),1.56(q,J=9.5Hz,4H); MS(ESI)m / z:calculated for C 24 H 20 Cl2N4O4[M+H] + :499.09,found:499.0926.

[0067] Example 5

[0068] Synthesis of intermediate ethyl 5-(4-chlorobenzyl)-1,3,4-oxadiazole-2-carboxylate

[0069]

[0070] Step 1: Synthesis of 2-(4-chlorophenyl)acetylhydrazine

[0071] Following the method described in step 1 of Example 3, 192 mg of white solid was obtained, with a yield of 96.00%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 9.23 (s, 1H), 7.38-7.33 (m, 2H), 7.30-7.25 (m, 2H), 4.22 (d, J = 3.2Hz, 2H), 3.35 (s, 2H).

[0072] Step 2: Synthesis of ethyl 2-(2-(2-(4-chlorophenyl)acetyl)hydrazino)-2-oxoethyl acetate

[0073] Following the method described in step 2 of Example 3, 1.29 g of white solid was obtained, with a yield of 68.30%. 1 H NMR(300MHz,DMSO-d6)δ(ppm):10.81(s,1H),10.30(s,1H),7.42 -7.36(m,2H),7.35-7.29(m,2H),4.26(q,J=7.1Hz,2H),3.52(s,2H),1.28(t,J=7.1Hz,3H).

[0074] Step 3: Synthesis of ethyl 5-(4-chlorobenzyl)-1,3,4-oxadiazole-2-carboxylate

[0075] Following the method described in step 3 of Example 3, 821 mg of white solid was obtained, with a yield of 68.47%. 1 H NMR (300MHz, Chloroform-d) δ7.37-7.33(m,2H),7.31-7.27(m,3H),4.51(q,J=7.1Hz,2H),4.28(s,2H),1.45(t,J=7.1Hz,3H).

[0076] Example 6

[0077] Synthesis of trans-N-(4-(5-chlorobenzofuran-2-carbamate)cyclohexyl)-5-(4-chlorobenzyl)-1,3,4-oxadiazole-2-carboxamide (I-2)

[0078]

[0079] Step 1: Synthesis of 5-(4-chlorobenzyl)-1,3,4-oxadiazole-2-carboxylic acid

[0080] Following the method described in step 2 of Example 1, 138 mg of a white solid was obtained, with a yield of 78.23%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 10.25 (s, 1H), 7.41-7.35 (m, 2H), 7.35-7.28 (m, 2H).

[0081] Step 2: Synthesis of trans-N-(4-(5-chlorobenzofuran-2-carbamoyl)cyclohexyl)-5-(4-chlorobenzyl)-1,3,4-oxadiazole-2-carboxamide (I-2)

[0082] Following the method described in step 2 of Example 4, 48 mg of white solid was obtained, with a yield of 55.78%. 1 H NMR(400MHz,DMSO-d6)δ(ppm):9.23(d,J=8.2Hz,1H),8.66(d,J=8.1Hz,1H),7 .87(d,J=2.2Hz,1H),7.70(d,J=8.9Hz,1H),7.53(d,J=0.9Hz,1H),7.49(dd,J =8.9,2.2Hz,1H),7.47-7.42(m,2H),7.42-7.35(m,2H),4.39(s,2H),3.75(s, 2H),1.86(t,J=10.9Hz,4H),1.52(q,J=11.3Hz,4H); MS(ESI)m / z:calculated for C 25 H 22 Cl2N4O4[M+H] + :513.10,found:513.3..

[0083] Example 7

[0084] Synthesis of intermediate ethyl 5-(4-bromophenyl)-1,3,4-oxadiazole-2-carboxylate

[0085]

[0086] Step 1: Synthesis of 4-bromobenzoylhydrazine

[0087] Following the method described in step 1 of Example 3, 267 mg of white solid was obtained, with a yield of 53.40%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 9.87 (s, 1H), 7.79-7.74 (m, 2H), 7.69-7.64 (m, 2H), 4.53 (s, 2H).

[0088] Step 2: Synthesis of ethyl 2-(2-(4-bromobenzoyl)hydrazino)-2-oxoethyl acetate

[0089] Following the method described in step 2 of Example 3, 252 mg of a white solid was obtained, with a yield of 85.99%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 10.99 (s, 1H), 10.75-10.68 (m, 1H), 7.85-7.80 (m, 2H), 7.78-7.73 (m, 2H), 4.31 (q, J = 7.1Hz, 2H), 1.31 (t, J = 7.1Hz, 3H).

[0090] Step 3: Synthesis of ethyl 5-(4-bromophenyl)-1,3,4-oxadiazole-2-carboxylate

[0091] Following the method described in step 3 of Example 3, 153 mg of white solid was obtained, with a yield of 65.70%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.05-7.98 (m, 2H), 7.90-7.84 (m, 2H), 4.46 (q, J = 7.1Hz, 2H), 1.37 (t, J = 7.1Hz, 3H).

[0092] Example 8

[0093] Synthesis of trans-5-(4-bromophenyl)-N-(4-(5-chlorobenzofuran-2-carbamoyl)cyclohexyl)-1,3,4-oxadiazole-2-carboxamide (I-3)

[0094]

[0095] Step 1: Synthesis of 5-(4-bromophenyl)-1,3,4-oxadiazole-2-carboxylic acid

[0096] Following the method described in step 2 of Example 1, 102 mg of a white solid was obtained, with a yield of 73.62%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 9.39 (s, 1H), 8.00-7.95 (m, 2H), 7.86 -7.81 (m, 2H).

[0097] Step 2: Synthesis of trans-5-(4-bromophenyl)-N-(4-(5-chlorobenzofuran-2-carbamoyl)cyclohexyl)-1,3,4-oxadiazole-2-carboxamide (I-3)

[0098] Following the method described in step 2 of Example 4, 85 mg of a white solid was obtained, with a yield of 27.31%. 1 H NMR (300MHz, DMSO-d6) δ (ppm) 9.30 (d, J = 8.2Hz, 1H), 8.68 (d, J = 8.1Hz, 1H), 8.09-8.00 (m, 2H), 7.93-7.83 (m, 3H), 7.71 (d, J = 8.8Hz, 1H), 7.5 4(d,J=0.9Hz,1H),7.49(dd,J=8.8,2.2Hz,1H),3.79(d,J=8.3Hz,2H),1.91(d,J=9.1Hz,4H),1.56(q,J=9.7Hz,4H); MS(ESI)m / z:calculated for C 24H 20 BrClN4O4[M+H] + :543.04,found:543.0428..

[0099] Example 9

[0100] Synthesis of intermediate ethyl 5-(4-chloro-3-fluorophenyl)-1,3,4-oxadiazole-2-carboxylate

[0101]

[0102] Step 1: Synthesis of 3-fluoro-4-chlorobenzoylhydrazide

[0103] Following the method described in step 1 of Example 3, 1.73 g of white solid was obtained, with a yield of 91.88%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 9.95 (s, 1H), 7.85-7.76 (m, 1H), 7.75-7.66 (m, 2H), 4.47 (s, 2H).

[0104] Step 2: Synthesis of ethyl 2-(2-(4-chloro-3-fluorobenzoyl)hydrazino)-2-oxoethyl acetate

[0105] Following the method described in step 2 of Example 3, 540 mg of white solid was obtained, with a yield of 69.86%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 10.91 (s, 2H), 7.90-7.84 (m, 1H), 7.83-7.76 (m, 2H), 4.31 (q, J = 7.1Hz, 2H), 1.31 (t, J = 7.1Hz, 3H).

[0106] Step 3: Synthesis of ethyl 5-(4-chloro-3-fluorophenyl)-1,3,4-oxadiazole-2-carboxylate

[0107] Following the method described in step 3 of Example 3, 376 mg of a white solid was obtained, with a yield of 74.26%. 1 H NMR (300MHz, Chloroform-d) δ (ppm): 8.00-7.92 (m, 2H), 7.66 -7.59 (m, 1H), 4.58 (q, J = 7.2Hz, 2H), 1.51 (t, J = 7.1Hz, 3H).

[0108] Example 10

[0109] Synthesis of trans-5-(4-chloro-3-fluorophenyl)-N-(4-(5-chlorobenzofuran-2-carbamoyl)cyclohexyl)-1,3,4-oxadiazole-2-carboxamide (I-4)

[0110]

[0111] Step 1: Synthesis of 5-(4-chloro-3-fluorophenyl)-1,3,4-oxadiazole-2-carboxylic acid

[0112] Following the method described in step 2 of Example 1, 243 mg of a white solid was obtained, with a yield of 70.38%. 1 H NMR (300MHz, Methanol-d4) δ (ppm): 8.05-7.94 (m, 2H), 7.74 (dd, J = 8.4, 7.4Hz, 1H).

[0113] Step 2: Synthesis of trans-5-(4-chloro-3-fluorophenyl)-N-(4-(5-chlorobenzofuran-2-carbamoyl)cyclohexyl)-1,3,4-oxadiazole-2-carboxamide (I-4)

[0114] Following the method described in step 2 of Example 4, 104 mg of a white solid was obtained, with a yield of 29.43%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 9.28 (d, J = 8.1 Hz, 1H), 8.65 ( d, J = 8.1 Hz, 1H), 8.11 ( dd, J = 9.6, 1.9 Hz, 1H), 7.97 ( dd, J = 8.5, 1.9 Hz, 1H), 7.93 -7.86(m,2H),7.71(d,J=8.8Hz,1H),7.54(d,J=0.9Hz,1H),7.49(dd,J=8.9,2.2Hz,1H ),3.81(s,2H),1.92(d,J=8.9Hz,4H),1.56(q,J=9.8Hz,4H); MS(ESI)m / z:calculated for C 24 H 19 ClFN4O4[M+H] + :517.08,found:517.1..

[0115] Example 11

[0116] Synthesis of intermediate 3-(4-chlorophenyl)-1,2,4-oxadiazole-5-carboxylic acid

[0117]

[0118] Step 1: Synthesis of (Z)-4-chloro-N'-hydroxybenzamide

[0119] 4-Chlorobenzonitrile (0.30 g, 2.18 mmol) was dissolved in methanol (10 mL), hydroxylamine hydrochloride (227 mg, 3.27 mmol) and sodium carbonate (173 mg, 1.64 mmol) were dissolved in water (1 mL) and added to the methanol solution of 4-chlorobenzonitrile. The mixture was heated to 65 °C and reacted for 15 h. The reaction solution was concentrated under reduced pressure, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 337 mg of crude yellow solid, with a yield of 90.58%. 1 H NMR (400MHz, DMSO-d6) δ (ppm): 9.85 (s, 1H), 7.88-7.80 (m, 2H), 7.58-7.50 (m, 2H), 4.51 (s, 2H).

[0120] Step 2: Synthesis of ethyl 3-(4-chlorophenyl)-1,2,4-oxadiazole-5-carboxylate

[0121] (Z)-4-chloro-N'-hydroxybenzamide (337 mg, 1.98 mmol) was dissolved in tetrahydrofuran, and N,N-diisopropylethylamine (412 μl, 2.96 mmol) was added. Oxaloyl chloride monoethyl ester (405 mg, 2.96 mmol) was then slowly added, and the mixture was heated to 70 °C and reacted for 2 h. The reaction solution was concentrated under reduced pressure, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 150 / 1 to 50 / 1) to give 331 mg of a white solid, with a yield of 66.32%. 1 HNMR (300MHz, DMSO-d6) δ (ppm): 8.13-8.05 (m, 2H), 7.77-7.64 (m, 2H), 4.47 (q, J = 7.1Hz, 2H), 1.38 (t, J = 7.1Hz, 3H).

[0122] Step 3: Synthesis of 3-(4-chlorophenyl)-1,2,4-oxadiazole-5-carboxylic acid

[0123] Following the method described in step 2 of Example 1, 147 mg of white solid was obtained, with a yield of 49.96%. 1 H NMR (300MHz, Methanol-d4) δ (ppm): 8.15-8.09 (m, 2H), 7.60-7.54 (m, 2H).

[0124] Example 12

[0125] Synthesis of trans-N-(4-(5-chlorobenzofuran-2-carbamoyl)cyclohexyl)-3-(4-chlorophenyl)-1,2,4-oxadiazole-5-carboxamide (I-5)

[0126]

[0127] Following the method described in step 2 of Example 4, 107 mg of a white solid was obtained, with a yield of 42.39%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 9.42 (d, J = 8.2Hz, 1H), 8.66 (d, J = 8.1Hz, 1H), 8.13-8.06 (m, 2H), 7.88 (d, J = 2.2Hz, 1H), 7.70 (dt, J = 9.2, 2.4Hz, 3H),7.54(d,J=0.9Hz,1H),7.49(dd,J=8.8,2.2Hz,1H),3.87-3.74(m,2H),1.96-1.85(m,4H),1.56(q,J=9.7Hz,4H); MS(ESI)m / z:calculated for C 24 H 20 Cl2N4O4[M+H] + :499.09,found:499.0934.

[0128] Example 13

[0129] Synthesis of trans-5-chloro-N-(4-(((5-(4-chlorophenyl)-1,3,4-oxadiazol-2-yl)methyl)amino)cyclohexyl)benzofuran-2-carboxamide (I-6)

[0130]

[0131] Step 1: Synthesis of 4-chloro-N'-(2-chloroacetyl)benzoylhydrazide

[0132] 4-Chlorobenzoylhydrazide (1.92 g, 11.28 mmol) was dissolved in dichloromethane (20 mL), followed by the addition of DIPEA (2.92 g, 22.56 mmol) and chloroacetyl chloride (1.35 mL, 16.92 mmol). The reaction mixture was reacted at room temperature for 50 min. The reaction solution was concentrated under reduced pressure, washed with water by stirring, filtered, and the filter cake was washed with petroleum ether / ethyl acetate, filtered, and dried to give 2.11 g of a white solid, with a yield of 75.72%. 1H NMR (300MHz, DMSO-d6) δ (ppm): 10.64 (s, 1H), 10.42 (s, 1H), 7.90 (d, J = 8.3Hz, 2H), 7.60 (d, J = 8.2Hz, 2H), 4.22 (s, 2H).

[0133] Step 2: Synthesis of 2-(chloromethyl)-5-(4-chlorophenyl)-1,3,4-oxadiazole

[0134] 4-Chloro-N'-(2-chloroacetyl)benzoylhydrazide (1.78 g, 7.20 mmol) was dissolved in acetonitrile (15 mL), and POCl3 (2.01 mL, 21.59 mmol) was added. The tube was sealed, and the reaction mixture was heated to 85 °C and reacted for 10 h. Water was slowly added, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 80 / 1) to give 856 mg of a white solid, with a yield of 51.93%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.09-8.00 (m, 2H), 7.77-7.67 (m, 2H), 5.16 (s, 2H).

[0135] Step 3: Synthesis of trans-5-chloro-N-(4-(((5-(4-chlorophenyl)-1,3,4-oxadiazol-2-yl)methyl)amino)cyclohexyl)benzofuran-2-carboxamide (I-6)

[0136] 2-(chloromethyl)-5-(4-chlorophenyl)-1,3,4-oxadiazole (132 mg, 450 μmol) was dissolved in N,N-dimethylformamide (10 mL), and potassium carbonate (124 mg, 900 μmol) was added. The mixture was stirred at room temperature for 10 min, followed by the slow addition of N-(4-aminocyclohexyl)-5-chlorobenzofuran-2-carboxamide (103 mg, 450 μmol). The reaction was carried out at room temperature for 4 h. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 100 / 1) to give 67 mg of a white solid, with a yield of 30.70%. 1H NMR (300MHz, DMSO-d6) δ (ppm): 8.57 (d, J = 8.1Hz, 1H), 8.05-7.99 (m, 2H), 7.86 (d, J = 2.2Hz, 1H), 7.70 (dt, J = 8.9, 2.4Hz, 3H), 7.51 (d, J = 0.9Hz, 1H), 7. 47(dd,J=8.9,2.2Hz,1H),4.05(s,2H),3.81-3.70(m,1H),2.44(d,J=10.7 Hz,1H),2.00-1.91(m,2H),1.83(d,J=12.3Hz,2H),1.47-1.34(m,2H),1.21 -1.10(m,2H);MS(ESI)m / z:calculated for C 24 H 22 Cl2N4O3[M+H] + :485.11,found:485.1133..

[0137] Example 14

[0138] Synthesis of trans-5-chloro-N-(4-(((5-(4-chlorobenzyl)-1,3,4-oxadiazol-2-yl)methyl)amino)cyclohexyl)benzofuran-2-carboxamide (I-7)

[0139]

[0140] Step 1: Synthesis of 2-chloro-N'-(2-(4-chlorophenyl)acetyl)acetylhydrazine

[0141] Following the method described in step 1 of Example 13, 710 mg of a pale yellow solid was obtained, with a yield of 83.67%. 1 H NMR (300MHz,, DMSO-d6) δ (ppm): 10.38-10.29 (m, 2H), 7.41-7.37 (m, 2H), 7.32 (d, J = 8.5Hz, 2H), 4.14 (s, 2H), 3.51 (s, 2H).

[0142] Step 2: Synthesis of 2-(4-chlorobenzyl)-5-(chloromethyl)-1,3,4-oxadiazole

[0143] Following the method described in step 2 of Example 13, 282 mg of white solid was obtained, with a yield of 59.63%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 7.47-7.41 (m, 2H), 7.40-7.35 (m, 2H), 5.03 (s, 2H), 4.35 (s, 2H).

[0144] Step 3: Synthesis of trans-5-chloro-N-(4-(((5-(4-chlorobenzyl)-1,3,4-oxadiazol-2-yl)methyl)amino)cyclohexyl)benzofuran-2-carboxamide (I-7)

[0145] Following the method described in step 3 of Example 13, 89 mg of white solid was obtained, with a yield of 29.08%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.55 (d, J = 8.1Hz, 1H), 7.85 (d, J = 2.2Hz, 1H), 7.68 (d, J = 8.8Hz, 1H), 7.51 (d, J = 1.0Hz, 1H), 7.49 -7.40 (m, 3H), 7.37 -7.31(m,2H),4.28(s,2H),3.90(s,2H),3.77-3.65(m,1H),2.33(d,J=11.0Hz,1H),1.9 0-1.75(m,4H),1.36(q,J=11.9Hz,2H),1.07(q,J=11.4Hz,2H); MS(ESI)m / z:calculated for C 24 H 22 Cl2N4O3[M+H] + :499.12,found:499.1303.

[0146] Example 15

[0147] Synthesis of trans-N-(4-(((5-(4-bromophenyl)-1,3,4-oxadiazol-2-yl)methyl)amino)cyclohexyl)-5-chlorobenzofuran-2-carboxamide (I-8)

[0148]

[0149] Step 1: Synthesis of 4-bromo-N'-(2-chloroacetyl)benzoylhydrazide

[0150] Following the method described in step 1 of Example 13, 163 mg of white solid was obtained, with a yield of 45.03%.

[0151] Step 2: Synthesis of 2-(4-bromophenyl)-5-(chloromethyl)-1,3,4-oxadiazole

[0152] Following the method described in step 2 of Example 13, 86 mg of white solid was obtained, with a yield of 89.87%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.00-7.92 (m, 2H), 7.88-7.79 (m, 2H), 5.15 (s, 2H).

[0153] Step 3: Synthesis of trans-N-(4-(((5-(4-bromophenyl)-1,3,4-oxadiazol-2-yl)methyl)amino)cyclohexyl)-5-chlorobenzofuran-2-carboxamide (I-8)

[0154] Following the method described in step 3 of Example 13, 108 mg of white solid was obtained, with a yield of 29.34%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.58 (d, J = 8.1Hz, 1H), 7.99-7.92 (m, 2H), 7.89 -7.81(m,3H),7.69(d,J=8.8Hz,1H),7.52(d,J=0.9Hz,1H),7.47(dd,J=8.9,2.2Hz,1H),4.05(s,2H),3.81-3.69(m,1H),2.44 (d,J=10.7Hz,1H),2.00-1.91(m,2H),1.83(d,J=12.4Hz,2H),1.45-1.32(m,2H),1.24-1.14(m,2H); MS(ESI)m / z:calculated for C 24 H 22 BrClN4O3[M+H] + :529.06,found:529.0640..

[0155] Example 16

[0156] Synthesis of trans-5-chloro-N-(4-(((5-(4-chloro-3-fluorophenyl)-1,3,4-oxadiazol-2-yl)methyl)amino)cyclohexyl)benzofuran-2-carboxamide (I-9)

[0157]

[0158] Step 1: Synthesis of 4-chloro-N'-(2-chloroacetyl)-3-fluorobenzoylhydrazine

[0159] Following the method described in step 1 of Example 13, 613 mg of white solid was obtained, with a yield of 86.19%.

[0160] Step 2: Synthesis of 2-(4-chloro-3-fluorophenyl)-5-(chloromethyl)-1,3,4-oxadiazole

[0161] Following the method described in step 2 of Example 13, 306 mg of white solid was obtained, with a yield of 53.56%. 1H NMR (300MHz, Chloroform-d) δ (ppm): 7.92-7.83 (m, 2H), 7.64 -7.55 (m, 1H), 4.81 (s, 2H).

[0162] Step 3: Synthesis of trans-5-chloro-N-(4-(((5-(4-chloro-3-fluorophenyl)-1,3,4-oxadiazol-2-yl)methyl)amino)cyclohexyl)benzofuran-2-carboxamide (I-9)

[0163] Following the method described in step 3 of Example 13, 84 mg of a white solid was obtained, with a yield of 27.30%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.59 (d, J = 8.1 Hz, 1H), 8.04-7.97 (m, 1H), 7.87 (dd, J = 3.7 ,1.4Hz,3H),7.69(d,J=8.8Hz,1H),7.51(d,J=0.9Hz,1H),7.47(dd,J=8.8,2.2Hz,1H),4. 06(s,2H),3.75(dt,J=7.8,3.8Hz,1H),2.44(d,J=10.9Hz,1H),1.96(d,J=12.6Hz,2H),1. 83(d,J=11.9Hz,2H),1.48-1.36(m,2H),1.14(d,J=13.0Hz,2H); MS(ESI)m / z:calculated for C 24 H 21 Cl2FN4O3[M+H] + :503.10,found:503.2..

[0164] Example 17

[0165] Synthesis of intermediate (1-(5-chlorobenzofuran-2-carbamate)piperidin-4-yl)tert-butyl carbamate

[0166]

[0167] Step 1: Synthesis of tert-butyl (1-nitrosopiperidin-4-yl)carbamate

[0168] N-Boc-aminopiperidine (300 mg, 1.50 mmol) was dissolved in tetrahydrofuran (10 mL), and 1 mol / L hydrochloric acid (2.70 mL, 2.70 mmol) was added dropwise under ice bath conditions. Then, sodium nitrite (248 mg, 3.59 mmol) dissolved in 5 mL of water was slowly added. The reaction was allowed to proceed for 3 h. The mixture was extracted with ethyl acetate and water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 315 mg of a white solid (91.72% yield). No further processing was required; it was used directly in the next reaction.

[0169] Step 2: Synthesis of (1-aminopiperidin-4-yl)tert-butyl carbamate

[0170] (1-Nitropiperidin-4-yl) tert-butyl carbamate (340 mg, 1.48 mmol) was dissolved in acetic acid / water (1:1, 15 mL), and zinc powder (383 mg, 5.93 mmol) was added. The reaction was maintained at 0 °C for 12 h. The reaction solution was filtered, the filtrate was concentrated, washed with ethyl acetate (30 mL), stirred for 30 min, filtered again, concentrated, and dried to obtain 251 mg of a clear oily liquid, with a yield of 78.62%. No further processing was required, and it was directly used in the next reaction.

[0171] Step 3: Synthesis of tert-butyl (1-(5-chlorobenzofuran-2-carbamate)piperidin-4-yl)carbamate

[0172] 5-Chlorobenzofuran-2-carboxylic acid (114 mg, 0.58 mmol) was dissolved in N,N-dimethylformamide (5 mL), followed by the addition of HATU (331 mg, 0.87 mmol) and N,N-diisopropylethylamine (0.30 mL, 1.74 mmol). The mixture was stirred at room temperature for 15 min, followed by the addition of tert-butyl (1-aminopiperidin-4-yl)carbamate (125 mg, 0.58 mmol). The reaction was allowed to proceed for 10 h. Water was added to the reaction mixture, and the mixture was stirred for 10 min. The mixture was then filtered, washed with a small amount of ethyl acetate, and yielded 123 mg of a white solid, with a yield of 53.79%. 1 H NMR (400MHz, DMSO-d6) δ (ppm): 9.77 (s, 1H), 7.95 (s, 1H), 7.86 (d, J = 2.2Hz, 1H), 7.70 (d, J = 8.8Hz, 1H), 7.53-7.45 (m, 2H), 6.85(d,J=7.8Hz,1H),2.96(d,J=10.5Hz,2H),2.78-2.66(m,4H),1.75(d,J=12.5Hz,2H),1.61-1.47(m,2H),1.39(s,9H).

[0173] Example 18

[0174] Synthesis of N-(1-(5-chlorobenzofuran-2-carbamoyl)piperidin-4-yl)-5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxamide (I-10)

[0175]

[0176] Step 1: Synthesis of N-(4-aminopiperidin-1-yl)-5-chlorobenzofuran-2-carboxamide

[0177] Following the method described in Example 2, 50 mg of a pale yellow solid was obtained, with a yield of 50.54%. 1 H NMR (300MHz, Chloroform-d) δ (ppm): 7.66 (d, J = 2.0Hz, 1H), 7.48 (d, J = 0.8Hz, 1H), 7.46-7.35 (m, 3H), 3.25 (dt, J = 10.6 ,3.9Hz,2H),2.86(dq,J=10.2,6.3,5.2Hz,1H),2.75(td,J=11.0,2.8Hz,2H),1.96(d,J=3.8Hz,2H),1.75-1.67(m,2H).

[0178] Step 2: Synthesis of N-(1-(5-chlorobenzofuran-2-carbamate)piperidin-4-yl)-5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxamide (I-10)

[0179] 5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxylic acid (39 mg, 0.17 mmol) was dissolved in N,N-dimethylformamide (5 mL), followed by the addition of HATU (84 mg, 0.22 mmol) and N-(4-aminopiperidin-1-yl)-5-chlorobenzofuran-2-carboxamide (50 mg, 0.17 mmol). The mixture was stirred at room temperature for 5 min, followed by the addition of N,N-diisopropylethylamine (74 μL, 0.43 mmol). The reaction was allowed to proceed for 6 h. The mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane / methanol = 200 / 1 to 100 / 1) to give 21 mg of a white solid, with a yield of 24.66%. 1H NMR (400MHz, DMSO-d6) δ (ppm): 9.84 (s, 1H), 9.39 (d, J = 8.0Hz, 1H), 8.11 (d, J = 8.2Hz, 2H), 7.88 (s, 1H), 7.72 (t, J = 8. 1Hz,3H),7.55-7.45(m,2H),3.85(s,1H),3.06(d,J=10.3Hz,2H),2.83(s,2H),1.85(s,4H);MS(ESI)m / z:calculated for C 23 H 19 Cl2N5O4[M+H] + :500.08,found:500.1..

[0180] Example 19

[0181] Synthesis of N-(1-(5-chlorobenzofuran-2-carboxamide)piperidin-4-yl)-5-(4-chlorobenzyl)-1,3,4-oxadiazole-2-carboxamide (I-11)

[0182]

[0183] Following the method described in step 2 of Example 18, 34 mg of a white solid was obtained, with a yield of 26.60%. 1 H NMR (400MHz, DMSO-d6) δ (ppm): 9.82 (s, 1H), 9.31 (d, J = 8.1Hz, 1H), 7.87 (d, J = 2.2Hz, 1H), 7.70 (d, J = 8.9Hz, 1H), 7.56- 7.46(m,3H),7.46-7.42(m,2H),7.39(d,J=8.5Hz,2H),4.39(s,2H),3.78(q,J=7.8Hz,1H),3.02(d,J=10.9Hz,2H),2.84 -2.75(m,2H),1.80(s,4H);MS(ESI)m / z:calculated for C 24 H 21 Cl2N5O4[M+H] + :514.10,found:514.5..

[0184] Example 20

[0185] Synthesis of intermediate ((1-(5-chlorobenzofuran-2-carbamate)piperidin-4-yl)methyl)tert-butyl carbamate

[0186]

[0187] Step 1: Synthesis of tert-butyl ((1-nitrosopiperidin-4-yl)methyl)carbamate

[0188] Following the method described in step 1 of Example 17, 876 mg of a white solid was obtained, with a yield of 96.45%. No further processing was required; it was directly used in the next reaction.

[0189] Step 2: Synthesis of tert-butyl ((1-aminopiperidin-4-yl)methyl)carbamate

[0190] Following the method described in step 2 of Example 17, 671 mg of a transparent oily liquid was obtained, with a yield of 81.27%. No further processing was required; it was directly used in the next reaction step.

[0191] Step 3: Synthesis of tert-butyl ((1-(5-chlorobenzofuran-2-carbamate)piperidin-4-yl)methyl)carbamate

[0192] Following the method described in step 3 of Example 17, 151 mg of white solid was obtained, with a yield of 42.45%. 1 H NMR (400MHz, DMSO-d6) δ (ppm): 9.73 (s, 1H), 7.86 (d, J = 2.2Hz, 1H), 7.70 (d, J = 8.8Hz, 1H), 7.53-7.45 (m, 2H), 6.87 (t, J = 5.8Hz, 1H),2.99(d,J=10.4Hz,2H),2.85(t,J=6.1Hz,2H),2.75-2.60(m,2H),1.65(d,J=12.8Hz,2H),1.39(s,9H),1.35-1.14(m,3H).

[0193] Example 21

[0194] Synthesis of N-((1-(5-chlorobenzofuran-2-carboxamide)piperidin-4-yl)methyl)-5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxamide (I-12)

[0195]

[0196] Step 1: Synthesis of N-(4-(aminomethyl)piperidin-1-yl)-5-chlorobenzofuran-2-carboxamide

[0197] Referring to the method described in Example 2, 97 mg of a white solid was obtained, with a yield of 85.70%. MS (ESI) m / z: calculated for C 15 H 18 ClN3O2[M+H] + :308.1,found:308.3.

[0198] Step 5: Synthesis of N-((1-(5-chlorobenzofuran-2-carboxamide)piperidin-4-yl)methyl)-5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxamide (I-12)

[0199] Following the method described in step 2 of Example 18, 41 mg of white solid was obtained, with a yield of 49.07%. 1 H NMR (400MHz, DMSO-d6) δ (ppm): 9.76 (s, 1H), 9.42 (t, J = 6.1Hz, 1H), 8.14-8.08 (m, 2H),7.86(d,J=2.2Hz,1H),7.75-7.68(m,3H),7.51(s,1H),7.48(dd,J=8.9,2.3H z,1H),3.24(t,J=6.4Hz,2H),3.03(d,J=10.3Hz,2H),2.70(t,J=10.9Hz,2H),1.7 5(d,J=12.9Hz,2H),1.65(s,1H),1.36(t,J=11.3Hz,2H); MS(ESI)m / z:calculated for C 24 H 21 Cl2N5O4[M+H] + :514.10,found:514.2..

[0200] Example 22

[0201] Synthesis of N-((1-(5-chlorobenzofuran-2-carbamoyl)piperidin-4-yl)methyl)-5-(4-chlorobenzyl)-1,3,4-oxadiazole-2-carboxamide (I-13)

[0202]

[0203] Following the method described in step 2 of Example 18, 34 mg of a white solid was obtained, with a yield of 39.61%. 1H NMR (300MHz, DMSO-d6) δ (ppm): 9.77 (s, 1H), 9.34 (t, J = 6.0 Hz, 1H), 7.87 (d, J = 2 .3Hz,1H),7.70(d,J=8.9Hz,1H),7.52-7.42(m,4H),7.39(d,J=8.6Hz,2H),4.39 (s,2H),3.17(t,J=6.3Hz,2H),3.00(d,J=10.8Hz,2H),2.76-2.61(m,2H),1.70( d,J=13.4Hz,2H),1.60(s,1H),1.33(d,J=14.8Hz,2H); MS(ESI)m / z:calculated for C 25 H 23 Cl2N5O4[M+H] + :528.11,found:528.5..

[0204] Example 23

[0205] Synthesis of 5-(4-bromophenyl)-N-((1-(5-chlorobenzofuran-2-carbamoyl)piperidin-4-yl)methyl)-1,3,4-oxadiazole-2-carboxamide (I-14)

[0206]

[0207] Following the method described in step 2 of Example 18, 48 mg of white solid was obtained, with a yield of 46.38%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 9.78 (s, 1H), 9.44 (t, J = 6.0Hz, 1H), 8.08-7. 99(m,2H),7.87(dt,J=6.6,2.5Hz,3H),7.70(d,J=8.9Hz,1H),7.52-7.45(m, 2H), 3.24 (t, J = 6.3Hz, 2H), 3.02 (d, J = 10.2Hz, 2H), 2.76-2.66 (m, 2H), 1.75 (d, J = 13.1Hz, 2H), 1.64 (s, 1H), 1.41-1.31 (m, 2H); MS (ESI) m / z: calculated for C 24 H 21 BrClN5O4[M+H] + :558.05,found:558.4..

[0208] Example 24

[0209] Synthesis of 5-(4-chloro-3-fluorophenyl)-N-((1-(5-chlorobenzofuran-2-carboxamide)piperidin-4-yl)methyl)-1,3,4-oxadiazole-2-carboxamide (I-15)

[0210]

[0211] Following the method described in step 2 of Example 18, 30 mg of a white solid was obtained, with a yield of 29.40%. 1 H NMR (400MHz, DMSO-d6) δ (ppm): 9.78 (s, 1H), 9.45 (t, J = 6.0Hz, 1H), 8.11 (dd, J = 9.5, 1.9Hz ,1H),7.96(dd,J=8.5,1.9Hz,1H),7.91(d,J=7.4Hz,1H),7.89-7.85(m,1H),7.70(d,J=8. 8Hz,1H),7.54-7.45(m,2H),3.24(t,J=6.5Hz,2H),3.02(d,J=10.3Hz,2H),2.74-2.65(m, 2H),1.75(d,J=12.9Hz,2H),1.65(s,1H),1.36(t,J=10.8Hz,2H); MS(ESI)m / z:calculated for C 24 H 20 Cl2FN5O4[M+H] + :532.09,found:532.4.

[0212] Example 25

[0213] Synthesis of N-(1-(5-chlorobenzofuran-2-carbonyl)piperidin-4-yl)-5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxamide (I-16)

[0214]

[0215] Step 1: Synthesis of tert-butyl (1-(5-chlorobenzofuran-2-carbonyl)piperidin-4-yl)carbamate

[0216] 5-Chlorobenzofuran-2-carboxylic acid (200 mg, 1.02 mmol) was dissolved in N,N-dimethylformamide (5 mL), followed by the sequential addition of HATU (774 mg, 2.03 mmol) and N,N-diisopropylethylamine (0.53 mL, 3.05 mmol). The mixture was stirred at room temperature for 15 min, followed by the addition of N-Boc aminopiperidine (214 mg, 1.07 mmol), and the reaction was allowed to proceed for 4.5 h. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to give 141 mg of a milky white solid, in 36.58% yield. 1 H NMR (300MHz, Chloroform-d) δ (ppm): 7.64 (dd, J=2.2, 0.6Hz, 1H), 7.47 (dt, J=8.8, 0.8Hz, 1H), 7.37 (dd, J=8.8, 2.1Hz, 1H), 7.23 (d, J=0.9 Hz,1H),6.82(d,J=8.1Hz,1H),4.52(s,2H),3.88-3.68(m,1H),3.35-2.92(m,2H),2.17-2.02(m,2H),1.67(d,J=17.8Hz,2H),1.47(s,9H).

[0217] Step 2: Synthesis of (4-aminopiperidin-1-yl)(5-chlorobenzofuran-2-yl)methyl ketone

[0218] Following the method described in Example 2, 62 mg of a white solid was obtained by purification by silica gel column chromatography (dichloromethane / methanol = 100 / 1), with a yield of 59.77%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 7.82 (d, J = 2.2Hz, 1H), 7.72 (d, J = 8.8Hz, 1H), 7.47 (dd, J = 8.9, 2.2Hz, 1H), 7.32 (s, 1H), 4.04 (s, 2H), 3.08 -2.82(m,3H),1.84-1.75(m,2H),1.28-1.18(m,2H).

[0219] Step 3: Synthesis of N-(1-(5-chlorobenzofuran-2-carbonyl)piperidin-4-yl)-5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxamide (II-7)

[0220] Following the method described in step 2 of Example 18, 145 mg of a white solid was obtained by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1), with a yield of 64.06%. 1H NMR (400MHz, DMSO-d6) δ (ppm): 9.38 (d, J = 8.0Hz, 1H), 8.15-8.08 (m, 2H), 7.84 (d,J=2.2Hz,1H),7.73(dd,J=8.8,2.7Hz,3H),7.48(dd,J=8.8,2.3Hz,1H),7. 37(s,1H),4.33-4.13(m,2H),3.03(s,1H),1.94(d,J=12.8Hz,2H),1.67(qd,J =12.2,11.5,3.6Hz,2H),1.25(q,J=5.5,4.0Hz,2H); MS(ESI)m / z:calculated forC 23 H 18 Cl2N4O4[M+H] + :485.07,found:485.4.

[0221] Example 26

[0222] Synthesis of N-((1-(5-chlorobenzofuran-2-carbonyl)piperidin-4-yl)methyl)-5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxamide (I-17)

[0223]

[0224] Step 1: Synthesis of tert-butyl ((1-(5-chlorobenzofuran-2-carbonyl)piperidin-4-yl)methyl)carbamate

[0225] Following the method described in step 1 of Example 25, 176 mg of a pale yellow solid was obtained, with a yield of 41.92%. 1 H NMR(300MHz,Chloroform-d)δ(ppm):7.66-7.61(m,1H),7.47(dt,J=8.8,0.7Hz,1H),7.36(dd,J=8.8,2.1Hz,1H),7.20(d ,J=0.9Hz,1H),6.83(d,J=8.2Hz,1H),4.69(s,2H),3.09(s,3H),1.85(d,J=12.4Hz,3H),1.47(s,9H),1.37-1.26(m,2H).

[0226] Step 2: Synthesis of (4-(aminomethyl)piperidin-1-yl)(5-chlorobenzofuran-2-yl)methyl ketone

[0227] Following the method described in Example 2, 107 mg of a transparent oily liquid was obtained by silica gel column chromatography (dichloromethane / methanol = 75 / 1), with a yield of 81.59%.1 H NMR (300MHz, Chloroform-d) δ (ppm): 7.63 (dd, J=2.2, 0.6Hz, 1H), 7.47 (dt, J=8.8, 0.8Hz, 1H), 7.36 (dd, J=8.8, 2.1Hz, 1H), 7.20 (d, J=0.9Hz, 1H), 4.58 (d, J = 70.8Hz, 2H), 3.01 (d, J = 93.7Hz, 2H), 2.68 (d, J = 6.5Hz, 2H), 1.90 (d, J = 12.8Hz, 2H), 1.76 (s, 3H), 1.29 (dt, J = 9.6, 2.6Hz, 2H).

[0228] Step 3: Synthesis of N-((1-(5-chlorobenzofuran-2-carbonyl)piperidin-4-yl)methyl)-5-(4-chlorophenyl)-1,3,4-oxadiazole-2-carboxamide (I-17)

[0229] Following the method described in step 2 of Example 18, 34 mg of a white solid was obtained by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1), with a yield of 18.63%. 1 H NMR (400MHz, Chloroform-d) δ (ppm): 8.17-8.08 (m, 2H), 7.64 (d, J = 2.1Hz, 1H) ,7.60-7.52(m,2H),7.47(d,J=8.8Hz,1H),7.39-7.31(m,2H),7.23(d,J=0.9Hz ,1H),4.62(d,J=79.4Hz,2H),3.49(s,2H),2.97(d,J=67.1Hz,2H),2.09-2.01( m,1H),1.95(d,J=13.4Hz,2H),1.47-1.39(m,2H);MS(ESI)m / z:calculatedfor C 24 H 20 Cl2N4O4[M+H] + :499.09,found:499.2..

[0230] Example 27

[0231] Synthesis of 5-chloro-N-(1-(5-(4-chlorophenyl)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)benzofuran-2-carboxamide (I-18)

[0232]

[0233] Step 1: Synthesis of tert-butyl 4-(5-chlorobenzofuran-2-carboxamido)piperidine-1-carboxylic acid

[0234] Following the method described in step 1 of Example 25, 280 mg of a yellow solid was obtained, with a yield of 72.65%. 1 H NMR (300MHz, Chloroform-d) δ (ppm): 7.67 (dd, J=2.1, 0.6Hz, 1H), 7.48-7.42 (m, 2H), 7.39 (dd, J=8.7, 2.1Hz, 1H), 6.52 (d, J=8.2Hz, 1H), 4.17 (dddd, J=15.6, 11.8, 7.8, 4.3Hz, 3H), 3.02-2.90 (m, 2H), 2.05 (dd, J=12.9, 3.7Hz, 2H), 1.71 (d, J=6.7Hz, 2H), 1.47 (s, 9H).

[0235] Step 2: Synthesis of 5-chloro-N-(piperidin-4-yl)benzofuran-2-carboxamide

[0236] Following the method described in Example 2, 162 mg of a white solid was obtained by purification by silica gel column chromatography (dichloromethane / methanol = 50 / 1), with a yield of 78.64%. 1 H NMR(300MHz,DMSO-d6)δ(ppm):8.67(d,J=8.0Hz,1H),7.87(d,J=2.2Hz,1H),7 .70(d,J=8.9Hz,1H),7.54(d,J=0.9Hz,1H),7.48(dd,J=8.8,2.2Hz,1H),3.85( dtd,J=11.6,7.5,4.0Hz,1H),3.05-2.93(m,2H),2.57(dd,J=12.3,2.5Hz,2H) ,1.79-1.69(m,2H),1.48(qd,J=12.1,4.1Hz,2H),1.24(dd,J=6.9,2.6Hz,1H).

[0237] Step 3: Synthesis of 5-chloro-N-(1-(5-(4-chlorophenyl)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)benzofuran-2-carboxamide (I-18)

[0238] Following the method described in step 2 of Example 18, 154 mg of a pale yellow solid was obtained by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1), with a yield of 54.60%. 1H NMR (400MHz, DMSO-d6) δ (ppm): 8.80 (d, J = 8.0Hz, 1H), 8.11-8.05 (m, 2H), 7.88 (d,J=2.2Hz,1H),7.75-7.68(m,3H),7.56(d,J=0.9Hz,1H),7.49(dd,J=8.9,2 .3Hz,1H),4.52(dd,J=27.4,13.6Hz,2H),4.20(dtt,J=11.3,8.0,4.0Hz,1H), 3.49-3.41(m,1H),3.16-3.08(m,1H),1.97(td,J=16.5,15.8,3.9Hz,2H),1.73 -1.56(m,2H);MS(ESI)m / z:calculated for C 23 H 18 Cl2N4O4[M+H] + :485.07,found:485.2..

[0239] Example 28

[0240] Synthesis of 5-chloro-N-((1-(5-(4-chlorophenyl)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)methyl)benzofuran-2-carboxamide (I-19)

[0241]

[0242] Step 1: Synthesis of tert-butyl 4-((5-chlorobenzofuran-2-carboxamido)methyl)piperidine-1-carboxylic acid

[0243] Following the method described in step 1 of Example 25, 226 mg of a pale yellow solid was obtained, with a yield of 61.64%. 1 H NMR(400MHz,Chloroform-d)δ(ppm):7.69 -7.66(m,1H),7.46 -7.43(m,2H),7.39(dd,J=8.7,2.1Hz,1H),6.75(d,J=6.7Hz,1H),4.16(s,2H),3.41(s,2H),2.71(d,J=13.4Hz,2 H),1.85(ddd,J=10.4,7.0,3.5Hz,1H),1.78(d,J=13.8Hz,2H),1.47(s,9H),1.26(ddd,J=16.3,10.0,4.2Hz,2H).

[0244] Step 2: Synthesis of 5-chloro-N-(piperidin-4-ylmethyl)benzofuran-2-carboxamide

[0245] Following the method described in Example 2, 66 mg of a white solid was obtained, with a yield of 39.19%. 1 H NMR (300MHz, Chloroform-d) δ (ppm): 7.67 (d, J=1.9Hz, 1H), 7.47-7.41 (m, 2H), 7.39 (dd, J=8.8, 2.1Hz, 1H), 6.74 (s, 1H), 3.40 (t, J=6 .2Hz,2H),3.15(dt,J=12.3,3.4Hz,2H),2.64(td,J=12.2,2.3Hz,2H),2.16(s,2H),1.79(dt,J=12.3,3.5Hz,3H),1.33-1.28(m,1H).

[0246] Step 3: Synthesis of 5-chloro-N-((1-(5-(4-chlorophenyl)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)methyl)benzofuran-2-carboxamide (I-19)

[0247] Following the method described in step 2 of Example 18, 20 mg of a white solid was obtained by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1), with a yield of 17.77%. 1 H NMR (400MHz, Chloroform-d) δ (ppm): 8.13-8.08 (m, 2H), 7.68 (d, J = 2.0Hz, 1H), 7.57-7.51 (m, 2H), 7.44 (s, 2H),7.40(dd,J=8.9,2.1Hz,1H),6.80(t,J=6.3Hz,1H),5.03(ddt,J=13.7,4.5,2.4Hz,1H),4.81(ddt,J=1 3.5, 4.6, 2.4Hz, 1H), 3.46 (t, J=6.6Hz, 2H), 3.36-3.23 (m, 1H), 2.91 (td, J=12.9, 2.9Hz, 1H), 2.09 (dqd, J= 8.1,6.0,5.1,3.4Hz,1H),2.03-1.96(m,2H),1.44(tdd,J=13.1,8.7,4.9Hz,2H); MS(ESI)m / z:calculated for C 24 H 20 Cl2N4O4[M+H] + :499.09,found:499.2..

[0248] Example 29

[0249] Synthesis of the intermediate trans-(4-(6-chloroquinoline-2-carbamate)cyclohexyl)tert-butyl carbamate

[0250]

[0251] Step 1: Synthesis of 6-chloroquinoline-2-carboxaldehyde

[0252] 2-Methyl-6-chloroquinoline (200 mg, 1.13 mmol) was dissolved in 1,4-dioxane (5 mL), and tin dioxide (750 mg, 6.76 mmol) was added. The mixture was heated to 80 °C and reacted for 3 h. The solution was filtered through diatomaceous earth, concentrated under reduced pressure, and dried to give 198 mg of a pink solid, with a yield of 91.78%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 10.12 (d, J = 0.8Hz, 1H), 8.59 (d, J = 8.5Hz, 1H), 8.31 (d, J =2.4Hz, 1H), 8.26 (d, J = 9.0Hz, 1H), 8.05 (d, J = 8.5Hz, 1H), 7.94 (dd, J = 9.0, 2.4Hz, 1H).

[0253] Step 2: Synthesis of 6-chloroquinoline-2-carboxylic acid

[0254] 6-Chloroquinoline-2-carboxaldehyde (107 mg, 0.56 mmol) was dissolved in tert-butanol (10 mL), and then an aqueous solution (5 mL) containing sodium chlorite (152 mg, 1.68 mmol) and sodium dihydrogen phosphate (201 mg, 1.68 mmol) was slowly added dropwise. The reaction was allowed to proceed for 10 h. Sodium sulfite (228 mg, 2.23 mmol) was dissolved in 5 mL of water and then slowly added to the reaction solution to quench the reaction. After cooling to room temperature, 1 mol / L hydrochloric acid was slowly added dropwise to adjust the pH to 4–5. The mixture was filtered and dried to give 73 mg of a white solid, with a yield of 62.97%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.54 (d, J = 8.6Hz, 1H), 8.26 (d, J = 2.4Hz, 1H), 8.17 (dd, J = 8.8, 3.8Hz, 2H), 7.88 (dd, J = 9.0, 2.4Hz, 1H).

[0255] Step 3: Synthesis of trans-(4-(6-chloroquinoline-2-carbamate)cyclohexyl)tert-butyl carbamate

[0256] Following the method described in step 3 of Example 1, 102 mg of a white solid was obtained, with a yield of 71.82%. 1H NMR(400MHz, DMSO-d6)δ(ppm):8.68(d,J=8.5Hz,1H),8.54(d,J=8.5Hz,1H),8.25(d,J=2.4Hz,1H),8.18(dd,J=8.7,2.6Hz,2H),7.89(dd,J=9.0,2.4H z,1H),6.79(d,J=8.0Hz,1H),3.83-3.71(m,1H),3.25-3.20(m,1H),1.85( d, J=11.3Hz, 4H), 1.55 (q, J=12.5Hz, 2H), 1.39 (s, 9H), 1.31-1.23 (m, 2H).

[0257] Example 30

[0258] Synthesis of trans-5-(4-chlorophenyl)-N-(4-(6-chloroquinoline-2-carbamate)cyclohexyl)-1,3,4-oxadiazole-2-carboxamide (I-20)

[0259]

[0260] Step 1: Synthesis of trans-N-(4-aminocyclohexyl)-6-chloroquinoline-2-carboxamide

[0261] Following the method described in Example 2, 167 mg of a pale yellow solid was obtained, with a yield of 73.52%. 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.71 (d, J = 8.4Hz, 1H), 8.55 (d, J = 8.6Hz, 1H), 8.26 (d, J = 2.4Hz, 1H), 8.17 (t, J = 8.9Hz, 2H), 7.89(dd,J=9.0,2.5Hz,1H),3.82(dtd,J=12.0,8.2,4.3Hz,1H),3.08-2.95(m,1H),2.04-1.87(m,4H),1.64-1.39(m,4H).

[0262] Step 5: Synthesis of trans-5-(4-chlorophenyl)-N-(4-(6-chloroquinoline-2-carbamoyl)cyclohexyl)-1,3,4-oxadiazole-2-carboxamide (I-20)

[0263] Following the method described in step 3 of Example 1, 96 mg of a white solid was obtained, with a yield of 84.03%. 1H NMR (400MHz, DMSO-d6) δ (ppm): 9.29 (d, J = 8.2Hz, 1H), 8.71 (d, J = 8.5Hz, 1H), 8.55 (d, J = 8.6Hz, 1H), 8.26 (d, J = 2.4Hz, 1H), 8 .20(dd,J=8.8,4.6Hz,2H),8.15-8.08(m,2H),7.89(dd,J=9.0,2.4Hz,1H),7.76-7.70(m,2H),3.86(d,J=10.0Hz,2H),1.98 -1.89(m,4H),1.69-1.59(m,4H);MS(ESI)m / z:calculated for C 25 H 21 Cl2N5O3[M+H] + :510.10,found:510.5..

[0264] Example 31

[0265] Synthesis of trans-5-(4-chlorobenzyl)-N-(4-(6-chloroquinoline-2-carboxamido)cyclohexyl)-1,3,4-oxadiazole-2-carboxamide (I-21)

[0266]

[0267] Following the method described in step 3 of Example 1, 43 mg of white solid was obtained, with a yield of 49.82%. 1 H NMR (400MHz, DMSO-d6) δ (ppm): 9.21 (d, J = 8.3Hz, 1H), 8.69 (d, J = 8.5Hz, 1H), 8. 55(d,J=8.6Hz,1H),8.25(d,J=2.5Hz,1H),8.19(dd,J=8.8,4.9Hz,2H),7.88(dd ,J=9.0,2.5Hz,1H),7.47-7.42(m,2H),7.41-7.37(m,2H),4.39(s,2H),3.81(s ,2H),1.94-1.84(m,4H),1.60(q,J=11.9,11.1Hz,4H); MS(ESI)m / z:calculated for C 26 H 23 Cl2N5O3[M+H] + :524.12,found:524.5..

[0268] Example 32

[0269] Synthesis of trans-5-(4-bromophenyl)-N-(4-(6-chloroquinoline-2-carboxamido)cyclohexyl)-1,3,4-oxadiazole-2-carboxamide (I-22)

[0270]

[0271] Following the method described in step 3 of Example 1, 23 mg of a white solid was obtained, with a yield of 21.34%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.74 (d, J = 8.6Hz, 1H), 8.55 (d, J = 8.8Hz, 1H), 8.40 (d, J = 7.9Hz, 1H), 8.29-8.14 (m, 3H), 7.90 (d, J = 9.1Hz, 1H), 7.81 (d ,J=8.5Hz,2H),7.68(d,J=8.0Hz,2H),3.84(s,2H),1.92(d,J=11.5Hz,4H),1.65(d,J=12.5Hz,2H),1.49(d,J=12.3Hz,2H); MS(ESI)m / z:calculated for C 25 H 21 BrClN5O3[M+H] + :554.05,found:554.1..

[0272] Example 33

[0273] Synthesis of trans-5-(4-chloro-3-fluorophenyl)-N-(4-(6-chloroquinoline-2-carbamate)cyclohexyl)-1,3,4-oxadiazole-2-carboxamide (I-23)

[0274]

[0275] Following the method described in step 3 of Example 1, 47 mg of white solid was obtained, with a yield of 54.05%. 1H NMR(400MHz,DMSO-d6)δ(ppm):9.30(d,J=8.2Hz,1H),8.71(d,J=8.4Hz,1H),8 .55(d,J=8.5Hz,1H),8.25(d,J=2.4Hz,1H),8.20(dd,J=8.8,4.7Hz,2H),8.12( dd,J=9.5,1.9Hz,1H),7.97(dd,J=8.4,1.9Hz,1H),7.93-7.87(m,2H),3.94-3 .81(m,2H),1.94(d,J=8.3Hz,4H),1.70-1.59(m,4H); MS(ESI)m / z:calculated for C 25 H 20 Cl2FN5O3[M+H] + :528.09,found:528.4..

[0276] Example 34

[0277] Synthesis of trans-3-(4-chlorophenyl)-N-(4-(6-chloroquinoline-2-carbamoyl)cyclohexyl)-1,2,4-oxadiazole-5-carboxamide (I-24)

[0278]

[0279] Following the method described in step 3 of Example 1, 57 mg of white solid was obtained, with a yield of 53.85%. 1 H NMR (400MHz, DMSO-d6) δ (ppm): 9.43 (d, J = 8.2Hz, 1H), 8.71 (d, J = 8.5Hz, 1H), 8.55 (d, J = 8.6Hz, 1H), 8.26 (d, J = 2.4Hz, 1H), 8.20 (dd, J = 8.8, 4.6Hz, 2H), 8. 12-8.07(m,2H),7.89(dd,J=9.0,2.4Hz,1H),7.73-7.69(m,2H),3.86(s,2H),1.94(d,J=7.9Hz,4H),1.65(q,J=9.9,8.4Hz,4H); MS(ESI)m / z:calculated for C 25 H 21 Cl2N5O3[M+H] + :510.10,found:510.4.

[0280] Example 35

[0281] Synthesis of trans-6-chloro-N-(4-(((5-(4-chlorophenyl)-1,3,4-oxadiazol-2-yl)methyl)amino)cyclohexyl)quinoline-2-carboxamide (I-25)

[0282]

[0283] Following the method described in step 3 of Example 13, 34 mg of a white solid was obtained, with a yield of 49.36%. 1 H NMR(300MHz,DMSO-d6)δ(ppm):8.68(d,J=8.5Hz,1H),8.54(d,J=8.7Hz,1H),8.25(d,J =2.4Hz,1H),8.18(dd,J=8.8,3.0Hz,2H),8.05-7.99(m,2H),7.88(dd,J=9.1,2.4Hz,1H ),7.74-7.69(m,2H),5.33(s,2H),3.80(d,J=10.4Hz,1H),2.00(d,J=6.8Hz,1H),1.88( d,J=11.8Hz,4H),1.64-1.51(m,2H),1.36(t,J=11.9Hz,2H).;MS(ESI)m / z:calculated forC 25 H 23 Cl2N5O2[M+H] + :496.12,found:496.2..

[0284] Example 36

[0285] Synthesis of trans-6-chloro-N-(4-(((5-(4-chlorobenzyl)-1,3,4-oxadiazol-2-yl)methyl)amino)cyclohexyl)quinoline-2-carboxamide (I-26)

[0286]

[0287] Following the method described in step 3 of Example 13, 88 mg of white solid was obtained, with a yield of 65.47%. 1H NMR(300MHz,DMSO-d6)δ(ppm):8.62(d,J=8.5Hz,1H),8.54(d,J=8.6Hz,1H),8.25(d,J=2 .4Hz,1H),8.17(dd,J=8.8,6.2Hz,2H),7.88(dd,J=9.1,2.4Hz,1H),7.46-7.42(m,2H),7. 38-7.33(m,2H),4.29(s,2H),3.92(s,2H),3.80(d,J=11.5Hz,1H),2.41-2.32(m,1H),1.9 2-1.80(m,4H),1.51-1.40(m,2H),1.12(dd,J=14.8,4.5Hz,2H); MS(ESI)m / z:calculated for C 26 H 25 Cl2N5O2[M+H] + :510.14,found:510.5..

[0288] Example 37

[0289] Synthesis of trans-N-(4-(((5-(4-bromophenyl)-1,3,4-oxadiazol-2-yl)methyl)amino)cyclohexyl)-6-chloroquinoline-2-carboxamide (I-27)

[0290]

[0291] Following the method described in step 3 of Example 13, 73 mg of white solid was obtained, with a yield of 51.25%. 1 H NMR(400MHz, DMSO-d6)δ(ppm):8.65(d,J=8.5Hz,1H),8.54(d,J=8.5Hz,1H),8.24(d ,J=2.4Hz,1H),8.18(dd,J=8.8,5.8Hz,2H),7.96-7.92(m,2H),7.88(dd,J=9.1,2.4 Hz,1H),7.86-7.82(m,2H),5.33(s,2H),3.83-3.76(m,1H),2.07-1.93(m,1H),1.89 (d,J=11.7Hz,4H),1.62-1.53(m,2H),1.38-1.33(m,2H).;MS(ESI)m / z:calculated for C 25 H 23 BrClN5O2[M+H] + :540.07,found:540.8.

[0292] Example 38

[0293] Synthesis of trans-6-chloro-N-(4-(((5-(4-chloro-3-fluorophenyl)-1,3,4-oxadiazol-2-yl)methyl)amino)cyclohexyl)quinoline-2-carboxamide (I-28)

[0294]

[0295] Following the method described in step 3 of Example 13, 22 mg of white solid was obtained, with a yield of 21.66%. 1 H NMR(400MHz, DMSO-d6)δ(ppm):8.68(d,J=8.5Hz,1H),8.54(d,J=8.6Hz,1H),8.25(d,J=2.4Hz,1H),8.18(dd,J=8.8,4.1Hz,2H),8.04-7.99(m,1H),7.92 -7.86(m,3H),5.34(s,2H),3.80(d,J=9.0Hz,1H),2.06-1.95(m,1H),1.88(d,J=11 .5Hz,4H),1.58(q,J=11.8Hz,2H),1.36(t,J=11.2Hz,2H); MS(ESI)m / z:calculated for C 25 H 22 Cl2FN5O2[M+H] + :514.11,found:514.0..

[0296] Example 39

[0297] Synthesis of intermediate 2-(5-chlorobenzofuran-2-yl)-5-(piperidin-4-yl)-1,3,4-oxadiazole

[0298]

[0299] Step 1: Synthesis of 5-chlorobenzofuran-2-carbonylhydrazine

[0300] Following the method described in step 1 of Example 3, 248 mg of a white solid was obtained, with a yield of 95.15%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 10.13 (s, 1H), 7.87 (d, J = 2.2Hz, 1H), 7.69 (d, J = 8.8Hz, 1H), 7.52-7.43 (m, 2H), 4.63 (s, 2H).

[0301] Step 2: Synthesis of tert-butyl 4-(2-(5-chlorobenzofuran-2-carbonyl)hydrazine-1-carbonyl)piperidine-1-carboxylic acid

[0302] Following the method described in step 3 of Example 1, 180 mg of a pale yellow solid was obtained, with a yield of 95.60%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 10.68 (s, 1H), 10.03 (s, 1H), 7.92 (d, J = 2.2Hz, 1H), 7.74 (d, J = 8.8Hz, 1H), 7.64 (d, J = 0.9Hz, 1H), 7.52 (dd, J = 8.9,2.2Hz,1H),3.97(d,J=13.1Hz,2H),2.77(d,J=19.6Hz,2H),2.49-2.39(m,1H),1.73(d,J=12.8Hz,2H),1.54–1.43(m,2H),1.41(s,9H).

[0303] Step 3: Synthesis of 2-(5-chlorobenzofuran-2-yl)-5-(piperidin-4-yl)-1,3,4-oxadiazole

[0304] Following the method described in Example 2, 102 mg of a pale yellow solid was obtained, with a yield of 78.71%. MS (ESI) m / z: calculated for C 15 H 14 ClN3O2[M+H] + :304.08,found:304.43.

[0305] Example 39

[0306] Synthesis of 1-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)-2-(4-chlorophenoxy)ethane-1-one (II-1)

[0307]

[0308] Following the method described in step 3 of Example 1, 44 mg of a white solid was obtained after purification, with a yield of 27.74%. 1H NMR (300MHz, DMSO-d6) δ (ppm): 7.91 (d, J = 2.2Hz, 1H), 7.85 (d, J = 8.9Hz, 1H), 7.78 (s, 1H), 7.55 ( dd,J=8.9,2.2Hz,1H),7.37–7.30(m,2H),7.00–6.93(m,2H),4.90(d,J=4.0Hz,2H),4.26(d,J=13 .3Hz,1H),3.88(d,J=13.8Hz,1H),3.46(d,J=6.7Hz,1H),3.27(d,J=12.2Hz,1H),2.97(t,J=12. 2Hz,1H),2.13(s,2H),1.87(q,J=10.8Hz,1H),1.67(q,J=10.0Hz,1H).;MS(ESI)m / z:calculated for C 23 H 19 Cl2N3O4[M+H] + :472.08,found:472.4.

[0309] Example 40

[0310] Synthesis of trans-N-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl)-2-(4-chlorophenoxy)acetamide (II-2)

[0311]

[0312] Step 1: Synthesis of trans-4-(2-(5-chlorobenzofuran-2-carbonyl)hydrazine-1-carbonyl)cyclohexyl)tert-butyl carbamate

[0313] Following the method described in step 3 of Example 1, 160 mg of a pale yellow solid was obtained, with a yield of 80.53%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 10.65 (s, 1H), 9.92 (s, 1H), 7.92 (d, J = 2.2Hz, 1H), 7.74 (d, J = 8.9Hz, 1H), 7.63 (d, J = 0.9Hz, 1H), 7.51 (dd, J = 8.9, 2. 2Hz,1H),6.77(d,J=8.0Hz,1H),3.18(s,1H),2.16(t,J=12.1Hz,1H),1.8 1(t,J=12.4Hz,4H),1.38(d,J=2.7Hz,9H),1.21(dq,J=24.2,12.2Hz,4H).

[0314] Step 2: Synthesis of trans-4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl-1-amine

[0315] Following the method described in Example 2, 99 mg of a pale yellow solid was obtained, with a yield of 72.62%. MS (ESI) m / z: calculated for C 15 H 14 ClN3O2[M+H] + :318.09,found:318.06

[0316] Step 3: Synthesis of trans-N-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl)-2-(4-chlorophenoxy)acetamide (Ⅲ-2)

[0317] Following the method described in step 3 of Example 1, 57 mg of a milky white solid was obtained, with a yield of 37.62%. 1 H NMR(300MHz,DMSO-d6)δ(ppm):8.07(d,J=8.2Hz,1H),7.90(s,1H),7.84(d,J=8.9Hz,1H), 7.77(d,J=6.9Hz,1H),7.54(d,J=9.0Hz,1H),7.40-7.29(m,2H),6.98(dt,J=14.0,6.7Hz, 2H),4.49(s,2H),3.72(d,J=9.2Hz,1H),3.11-2.93(m,1H),2.19(d,J=13.5Hz,2H),1.91( d,J=12.3Hz,2H),1.67(d,J=12.4Hz,2H),1.47(q,J=12.6Hz,2H); MS(ESI)m / z:calculated for C 24 H 21 Cl2N3O4[M+H] + :486.09,found:486.4.

[0318] Example 41

[0319] Synthesis of trans-N-((4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl)methyl)-2-(4-chlorophenoxy)acetamide (II-3)

[0320]

[0321] Step 1: Synthesis of trans-((4-(2-(5-chlorobenzofuran-2-carbonyl)hydrazine-1-carbonyl)cyclohexyl)methyl)tert-butyl carbamate

[0322] Following the method described in step 3 of Example 1, 364 mg of a pale yellow solid was obtained, with a yield of 85.20%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 10.63 (s, 1H), 9.91 (s, 1H), 7.92 (d, J = 2.2Hz, 1H), 7.74 (d, J = 8.9Hz, 1H), 7.63 (d, J=0.9Hz,1H),7.51(dd,J=8.9,2.3Hz,1H),6.83(t,J=5.9Hz,1H),2.92-2.67(m,3H),2.19(t,J=12.0Hz,1H),1.86 -1.67(m,4H),1.38(s,9H),1.32(d,J=10.5Hz,2H),0.98-0.83(m,2H).

[0323] Step 2: Synthesis of trans-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl)methylamine

[0324] Following the method described in Example 2, 235 mg of a pale yellow solid was obtained, with a yield of 88.77%. MS (ESI) m / z: calculated for C 15 H 14 ClN3O2[M+H] + :332.11,found:332.92.

[0325] Step 3: Synthesis of trans-N-((4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl)methyl)-2-(4-chlorophenoxy)acetamide (II-3)

[0326] Following the method described in step 3 of Example 1, 108 mg of white solid was obtained, with a yield of 30.47%. 1H NMR(300MHz,DMSO-d6)δ(ppm):8.16(t,J=6.0Hz,1H),7.90(d,J=2.2Hz,1H),7.84(d,J=8.9H z,1H),7.76(d,J=1.0Hz,1H),7.54(dd,J=8.9,2.2Hz,1H),7.39-7.33(m,2H),7.03-6.97(m,2 H),4.52(s,2H),3.04(t,J=6.4Hz,2H),3.01-2.92(m,1H),2.15(d,J=11.7Hz,2H),1.79(d,J= 12.7Hz, 2H), 1.54 (dd, J=13.5, 10.1Hz, 3H), 1.07 (d, J=12.3Hz, 2H); MS (ESI) m / z: calculated for C 25 H 23 Cl2N3O4[M+H] + :500.11,found:500.4.

[0327] Example 42

[0328] Synthesis of trans-N-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl)-5-(4-chlorophenyl)-1,3,4-oxadiazol-2-carboxamide (II-4)

[0329]

[0330] Following the method described in step 3 of Example 1, 33 mg of a white solid was obtained, with a yield of 35.06%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 9.45 (t, J = 6.1Hz, 1H), 8.15-8.08 (m, 2H), 7.90 (d, J = 2.2Hz, 1H), 7.84 (d, J = 8.8Hz, 1H), 7.75 (d, J = 5.0Hz, 2H), 7.73 -7.70(m,1H),7.54(dd,J=8.9,2.2Hz,1H),3.21(d,J=6.5Hz,2H),3.04(s,1H),2.19(d,J=12.7Hz,2H) ,1.90(d,J=13.0Hz,2H),1.66(d,J=18.3Hz,1H),1.52(d,J=12.9Hz,2H);MS(ESI)m / z:calculatedfor C 26 H 21 Cl2N5O4[M+H] +:538.10,found:538.5..

[0331] Example 43

[0332] Synthesis of trans-N-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl)-3-(4-chlorophenyl)-1,2,4-oxadiazol-5-carboxamide (II-5)

[0333]

[0334] Following the method described in step 3 of Example 1, 30 mg of a white solid was obtained, with a yield of 23.11%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 9.56 (t, J = 6.1 Hz, 1H), 8.13-8.06 (m, 2H), 7.90 (d, J = 2.2H z,1H),7.84(d,J=8.9Hz,1H),7.76(d,J=0.9Hz,1H),7.74-7.65(m,2H),7.54(dd,J=8.9, 2.2Hz,1H),3.22(t,J=6.5Hz,2H),3.04(s,1H),2.19(d,J=12.7Hz,2H),1.90(d,J=13.0H z,2H),1.70(s,1H),1.55(t,J=12.9Hz,2H),1.21-1.13(m,2H); MS(ESI)m / z:calculated for C 26 H 21 Cl2N5O4[M+H] + :538.10,found:538.4..

[0335] Example 44

[0336] Synthesis of intermediate 4-(5-(5-chloro-1H-indol-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexane-1-amine

[0337]

[0338] Step 1: Synthesis of 5-chloro-1H-indole-2-carbonylhydrazide

[0339] Following the method described in step 1 of Example 39, 308 mg of a yellowish-white solid was obtained, with a yield of 91.33%. 1H NMR (400MHz, DMSO-d6) δ11.90–11.77(m,1H),9.88(s,1H),7.67(d,J=2.0Hz,1 H),7.43(d,J=8.7Hz,1H),7.17(dd,J=8.7,2.1Hz,1H),7.07(d,J=2.0Hz,1H).

[0340] Step 2: Synthesis of tert-butyl (4-(2-(5-chloro-1H-indole-2-carbonyl)hydrazine-1-carbonyl)cyclohexyl)carbamate

[0341] Following the method described in step 2 of Example 39, 408 mg of white solid was obtained, with a yield of 99.74%. 1 H NMR (300MHz, DMSO-d6) δ11.89(d,J=2.1Hz,1H),10.42(s,1H),9.86(s,1H),7.73(d,J=2.1Hz,1H),7.45(d,J=8.7Hz,1H),7.25–7.1 5(m,2H),6.77(d,J=8.1Hz,1H),3.10(m,1H)2.17(t,J=11.9Hz,1H),1.80(d,J=12.2Hz,4H),1.38(s,9H),1.38(m,2H),1.20(m,2H).

[0342] Step 3: Synthesis of 4-(5-(5-chloro-1H-indol-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexane-1-amine

[0343] Following the method described in step 3 of Example 39, 205 mg of a yellowish-white solid was obtained, with a yield of 86%. 1 H NMR(500MHz,Chloroform-d)δ9.61(s,1H),7.75(s,1H),7.63(s,1H),7.38(s,1H),7.06(s,1H ),5.69(s,1H),3.81(s,1H),3.24(s,1H),2.96(s,1H),1.80–1.69(m,4H),1.70–1.60(m,4H).

[0344] Example 45

[0345] Synthesis of N-(4-(5-(5-chloro-1H-indol-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl)-2-(4-chlorophenoxy)acetamide (II-6)

[0346]

[0347] Following the method described in step 3 of Example 1, 23 mg of a white solid was obtained, with a yield of 22.53%. 1 H NMR (300MHz, DMSO-d6) δ12.46(s,1H),8.07(d,J=8.0Hz,1H),7.74(d,J=2.0Hz,1H),7.48(d,J=8.8Hz,1H),7.37 (d,J=2.3Hz,1H),7.35(d,J=2.3Hz,1H),7.26(dd,J=8.7,2.1Hz,1H),7.15(s,1H),7.00(d,J=2.3Hz,1H),6.98( d,J=2.3Hz,1H),4.49(s,2H),3.71(ddd,J=11.6,7.6,3.8Hz,1H),3.00(tt,J=11.6,3.5Hz,1H),2.20(d,J=11.7 Hz,2H),1.91(dd,J=12.7,3.8Hz,2H),1.67(qd,J=13.0,3.1Hz,2H),1.57–1.37(m,2H).MS(ESI)m / z:calculated for C 24 H 22 Cl2N4O3[M+H] + :485.11,found:485.4.

[0348] Example 46

[0349] Synthesis of (4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)(5-(4-chlorophenyl)-1,3,4-oxadiazol-1-yl)methyl ketone (II-7)

[0350]

[0351] Following the method described in step 3 of Example 1, 54 mg of a white solid was obtained, with a yield of 40.00%. 1H NMR (300MHz, DMSO-d6) δ8.10(d,J=2.0Hz,1H),8.07(d,J=2.0Hz,1H),7.91(d,J=2.2Hz ,1H),7.85(d,J=8.9Hz,1H),7.79(d,J=0.9Hz,1H),7.74(d,J=2.0Hz,1H),7.72(d,J=2 .0Hz,1H),7.55(dd,J=8.9,2.2Hz,1H),4.50(dd,J=43.3,13.4Hz,2H),3.69–3.49(m,2 H),3.32–3.25(m,1H),2.31–2.16(m,2H),2.05–1.79(m,2H).MS(ESI)m / z:calculated forC 24 H 17 Cl2N5O4[M+H] + 510.07, found: 510.33.

[0352] Example 47

[0353] Synthesis of intermediate 2-(5-chloro-1H-indol-2-yl)-5-(piperidin-4-yl)-1,3,4-oxadiazole

[0354]

[0355] Step 1: Synthesis of tert-butyl 4-(2-(5-chloro-1H-indole-2-carbonyl)hydrazine-1-carbonyl)piperidine-1-carboxylate

[0356] The starting material was 1-(tert-butyloxycarbonyl)piperidine-4-carboxylic acid. Following the synthesis in step 3 of Example 1, 473 mg of a yellowish-white solid was obtained, with a yield of 99.72%. 1 H NMR (300MHz, DMSO-d6) δ11.90(d,J=2.1Hz,1H),10.46(s,1H),9.97(s,1H),7.74(d,J=2.0Hz,1H),7.45(d,J=8.7Hz,1H),7.26–7.1 6(m,2H),3.97(d,J=13.1Hz,2H),2.71(s,1H),1.81–1.69(m,2H),1.50(td,J=12.3,4.1Hz,2H),1.41(s,9H),1.23(t,J=8.2Hz,2H).

[0357] Step 2: Synthesis of 2-(5-chloro-1H-indol-2-yl)-5-(piperidin-4-yl)-1,3,4-oxadiazole

[0358] Following the synthesis in step 3 of Example 39, 156 mg of a brownish-yellow solid was obtained, with a yield of 91.92%. 1 H NMR (400MHz, DMSO-d6) δ11.93(s,1H),7.74(d,J=2.1Hz,1H),7.45(d,J=8.7Hz,1H), 7.23–7.18(m,2H),3.10(s,4H),2.85(s,1H),2.56(s,1H),1.88(d,J=17.2Hz,4H).

[0359] Example 48

[0360] Synthesis of (4-(5-(5-chloro-1H-indol-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)(5-(4-chlorophenyl)-1,3,4-oxadiazol-1-yl)methyl ketone (II-8)

[0361]

[0362] Following the method described in step 3 of Example 1, 53 mg of white solid was obtained, with a yield of 36.63%. 1 H NMR(500MHz,Chloroform-d)δ9.61(s,1H),8.14–8.08(m,3H),7.62(s,1H),7.56(s,1H),7.48–7.39(m,4H),7.17(s,1H),4.50( dd,J=43.3,13.4Hz,2H),3.69–3.49(m,2H),3.32–3.25(m,1H),2.31–2.16(m,2H),2.05–1.79(m,2H).MS(ESI)m / z:calculated for C 24 H 18 Cl2N6O3[M+H] + 509.08, found: 509.35.

[0363] Example 49

[0364] Synthesis of intermediate (1r,4r)-4-((5-(5-chloro-1H-indol-2-yl)-1,3,4-oxadiazol-2-yl)methyl)cyclohexane-1-amine

[0365]

[0366] Step 1: Synthesis of tert-butyl ((1r,4r)-4-(2-(2-(5-chloro-1H-indole-2-carbonyl)hydrazyl)-2-oxoethyl)cyclohexyl)carbamate

[0367] The starting materials were 5-chloro-1H-indole-2-carbonylhydrazide and (1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid, synthesized in step 1 of Example 44. Following the synthesis in step 3 of Example 1, 374 mg of a yellowish-white solid was obtained, with a yield of 78.72%. 1 H NMR (300MHz, DMSO-d6) δ11.94(s,1H),10.40(s,1H),9.87(s,1H),7.74(d,J=2.0Hz,1H),7.44(d,J=8.7Hz,1H),7.29–7.15(m,2H),6.70(d,J= 8.1Hz,1H),3.18(d,J=10.4Hz,1H),2.07(d,J=6.9Hz,2H),1.78(s,4H),1.62(m,1H),1.38(s,9H),1.19(d,J=12.6Hz,1H),1.16–0.92(m,3H).

[0368] Step 2: Synthesis of (1r,4r)-4-((5-(5-chloro-1H-indol-2-yl)-1,3,4-oxadiazol-2-yl)methyl)cyclohexane-1-amine

[0369] Following step 3 of Example 39, 48 mg of a brownish-yellow solid was obtained, with a yield of 30.27%. MS (ESI) m / z: calculated for C 17 H 19 ClN4O[M+H] + :330.82,found:330.83.

[0370] Example 50

[0371] Synthesis of N-((1r,4r)-4-((5-(5-chloro-1H-indol-2-yl)-1,3,4-oxadiazol-2-yl)methyl)cyclohexyl)-2-(4-chlorophenoxy)acetamide (II-9)

[0372]

[0373] Following the method described in step 3 of Example 1, 27 mg of a white solid was obtained, with a yield of 74.32%. 1H NMR (300MHz, DMSO-d6) δ12.45(s,1H),7.95(d,J=8.1Hz,1H),7.74(d,J=2.0Hz,1H) ,7.49(d,J=8.7Hz,1H),7.40–7.28(m,2H),7.26(dd,J=8.8,2.1Hz,1H),7.15(s,1H ),7.03–6.91(m,2H),4.45(s,2H),3.61(d,J=8.3Hz,1H),2.87(d,J=6.4Hz,2H),1. 80(d,J=8.3Hz,5H),1.40–1.23(m,2H),1.28–1.08(m,2H).MS(ESI)m / z:calculated for C 25 H 24 Cl2N4O3[M+H] + :499.12,found:499.39.

[0374] Example 51

[0375] Synthesis of intermediate (1r,4r)-4-((5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)methyl)cyclohexane-1-amine

[0376]

[0377] Step 1: Synthesis of tert-butyl ((1r,4r)-4-(2-(2-(5-chlorobenzofuran-2-carbonyl)hydrazino)-2-oxoethyl)cyclohexyl)carbamate

[0378] The starting materials were 5-chlorobenzofuran-2-carbonylhydrazide and (1r,4r)-4-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid, synthesized in step 1 of Example 39. Following the synthesis in step 3 of Example 1, 582 mg of a white solid was obtained, with a yield of 82.21%. 1 H NMR (300MHz, DMSO-d6) δ10.40(s,1H),9.87(s,1H),7.74(d,J=2.0Hz,1H),7.44(d,J=8.7Hz,1H),7.29–7.15(m,2H),6.70(d,J=8.1Hz, 1H),3.18(d,J=10.4Hz,1H),2.07(d,J=6.9Hz,2H),1.78(s,4H),1.62(m,1H),1.38(s,9H),1.19(d,J=12.6Hz,1H),1.16–0.92(m,3H).

[0379] Step 2: Synthesis of (1r,4r)-4-((5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)methyl)cyclohexane-1-amine.

[0380] Following the synthesis in step 3 of Example 39, 92 mg of a brownish-yellow solid was obtained, with a yield of 73%. 1 H NMR (300MHz, DMSO-d6) δ7.93–7.80(m,2H),7.76(s,1H),7.54(dd,J=8.8,2.2Hz,1H),2.87(d,J=6.4Hz,2H),1.75(s,5H),1.10(d,J=18.2Hz,5H).

[0381] Example 52

[0382] Synthesis of N-((1r,4r)-4-((5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)methyl)cyclohexyl)-2-(4-chlorophenoxy)acetamide (II-10)

[0383]

[0384] Following the method described in step 3 of Example 1, 53 mg of a white solid was obtained, with a yield of 84.32%. 1 H NMR (400MHz, DMSO-d6) δ7.93(d,J=8.1Hz,1H),7.89(d,J=2.2Hz,1H),7.84(d,J =8.8Hz,1H),7.75(d,J=0.9Hz,1H),7.54(dd,J=8.9,2.2Hz,1H),7.41–7.29(m,2 H),7.04–6.92(m,2H),3.61(dtt,J=11.5,7.8,3.9Hz,1H),2.90(d,J=6.5Hz,2H ),1.78(td,J=9.6,8.7,4.4Hz,5H),1.38–1.09(m,4H).MS(ESI)m / z:calculated for C 25 H 23 Cl2N3O4[M+H] + :500.11,found:500.39.

[0385] Example 53

[0386] Synthesis of intermediate 2-(4-chlorophenoxy)-N-((1r,4r)-4-(hydrazine carbonyl)cyclohexyl)acetamide

[0387]

[0388] Step 1: Synthesis of methyl (1r,4r)-4-(2-(4-chlorophenoxy)acetamido)cyclohexane-1-carboxylate

[0389] The starting material was methyl (1r,4r)-4-aminocyclohexane-1-carboxylate, and the method described in step 3 of Example 1 was followed. 335 mg of a white solid was obtained, with a yield of 72.24%. 1 H NMR(300MHz,DMSO-d6)δ7.97(d,J=7.8Hz,1H),7.40–7.29(m,2H),7.03–6.92(m,2H),4.45(s,2H),3.5 9(s,3H),2.25(ddt,J=11.6,7.1,3.5Hz,1H),2.00–1.85(m,2H),1.85–1.71(m,2H),1.49–1.13(m,5H).

[0390] Step 2: Synthesis of 2-(4-chlorophenoxy)-N-((1r,4r)-4-(hydrazine carbonyl)cyclohexyl)acetamide

[0391] Following the method described in step 3 of Example 1, 273 mg of a white solid was obtained, with a yield of 72.47%. 1 H NMR(300MHz, DMSO-d6)δ8.93(s,1H),7.95(d,J=8.1Hz,1H),7.44–7.26(m,2H),7.05–6.89(m,2H),4.45(s,2H),4.14 (s,2H),3.68–3.48(m,1H),2.00(tt,J=12.0,3.5Hz,1H),1.86–1.60(m,4H),1.54–1.34(m,2H),1.34–1.13(m,2H).

[0392] Example 54

[0393] Synthesis of 2-(4-chlorophenoxy)-N-((1r,4r)-4-(5-(6-chloroquinoline-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl)acetamide (II-11)

[0394]

[0395] Step 1: Synthesis of 2-(4-chlorophenoxy)-N-((1r,4r)-4-(2-(6-chloroquinoline-2-carbonyl)hydrazine-1-carbonyl)cyclohexyl)acetamide

[0396] The raw materials were 2-(4-chlorophenoxy)-N-((1r,4r)-4-(hydrazine carbonyl)cyclohexyl)acetamide from step 2 of Example 53 and 6-chloroquinoline-2-carboxylic acid from step 2 of Example 29. The method was described according to step 3 of Example 1. 261 mg of a white solid was obtained, with a yield of 72%. 1 H NMR (300MHz, DMSO-d6) δ10.60(s,1H),10.06(s,1H),8.58(d,J=8.6Hz,1H),8.28(d,J=2.4Hz,1H),8.22–8.09(m,2H),8.06–7.86(m,2H), 7.43–7.25(m,2H),7.07–6.88(m,2H),4.47(s,2H),3.72–3.53(m,1H),2.26(t,J=11.3Hz,1H),1.85(d,J=11.2Hz,4H),1.59–1.18(m,4H).

[0397] Step 2: Synthesis of 2-(4-chlorophenoxy)-N-((1r,4r)-4-(5-(6-chloroquinoline-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl)acetamide (II-11)

[0398] Following the synthesis in step 3 of Example 39, 92 mg of a white solid was obtained, with a yield of 20.00%. 1 H NMR(400MHz, DMSO-d6)δ8.61(d,J=8.7Hz,1H),8.38–8.26(m,2H),8.19(d,J=9.0Hz,1H),8.05(d, J=8.0Hz,1H),7.90(dd,J=9.0,2.4Hz,1H),7.44–7.30(m,2H),7.08–6.93(m,2H),4.49(s,2H),3.7 4(dtd,J=11.9,8.0,4.0Hz,1H),3.09(tt,J=12.0,3.7Hz,1H),2.28–2.17(m,2H),1.93(dd,J=13.2 ,3.7Hz,2H),1.71(qd,J=13.0,3.3Hz,2H),1.49(qd,J=12.8,3.6Hz,2H).MS(ESI)m / z:calculated for C 25 H 22 Cl2N4O3[M+H] + :497.11,found:497.39..

[0399] Example 55

[0400] Synthesis of intermediate 6-chloroquinoline-2-carboxamide

[0401]

[0402] Step 1: Synthesis of 6-bromoquinoline-2-formamide

[0403] 6-Bromoquinoline-2-carboxylic acid (1.49 g, 5.90 mmol) was dissolved in anhydrous acetonitrile (20 mL). Oxaloyl chloride (900 μl, 10.63 mmol) was added under ice bath conditions, followed by 5 drops of N,N-dimethylformamide. The reaction mixture was allowed to react for 3 h. The reaction mixture was then added to a solution of ammonia water (1.14 mL, 29.57 mmol):acetonitrile prepared under ice bath conditions at a ratio of 1:10. The reaction was allowed to proceed until complete after 20 min. The solution was filtered, dried, and yielded 1.16 g of a pale yellow solid, with a yield of 72.64%. 1 H NMR (300MHz, DMSO-d6) δ8.53(d,J=8.5Hz,1H),8.40(d,J=2.2Hz,1H),8.33(s,1H),8. 19(d,J=8.6Hz,1H),8.05(d,J=9.0Hz,1H),7.98(dd,J=9.0,2.2Hz,1H),7.84(s,1H).

[0404] Step 2: Synthesis of 6-bromoquinoline-2-nitrile

[0405] 6-Bromoquinoline-2-carboxamide (500 mg, 1.99 mmol) was dissolved in 10 mL of N,N-dimethylformamide, followed by the addition of phosphorus pentachloride (1.24 g, 5.97 mmol), and the reaction was carried out at 50 °C for 1 h. The mixture was then filtered with water. The filter cake was dissolved in dichloromethane, the filtrate was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1) to give a white solid in 33.83% yield. 1 H NMR (300MHz, DMSO-d6) δ8.64 (d, J=8.5Hz, 1H), 8.52–8.43 (m, 1H), 8.16–8.01 (m, 3H).

[0406] Step 3: Synthesis of 6-bromoquinoline-2-carboxamide

[0407] 6-Bromoquinoline-2-onitrile (220 mg, 943.93 μmol) was added to 10 mL of anhydrous methanol, followed by sodium methoxide (51 mg, 943.93 μmol), and the reaction was carried out at 40 °C for 30 min. NH₄Cl (151.47 mg, 2.83 mmol) was added, and the reaction was continued for 3 h. The pH was adjusted to approximately 8 using NH₃·H₂O. The mixture was filtered through diatomaceous earth and washed with a small amount of methanol. The filtrate was concentrated, and the residue was slurried with dichloromethane to precipitate a solid. The solid was then filtered to obtain a white solid with a yield of 86.51%. 1H NMR (300MHz, DMSO-d6) δ8.74(d,J=8.7Hz,1H),8.64–8.45(m,2H),8.12(d,J=9.0Hz,1H),8.08(dd,J=9.1,2.0Hz,1H),7.73(s,1H),6.26(s,2H).

[0408] Example 56

[0409] Synthesis of 2-(4-chlorophenoxy)-N-((1r,4r)-4-(5-(6-bromoquinolin-2-yl)-4H-1,2,4-triazol-3-yl)cyclohexyl)acetamide (II-12)

[0410]

[0411] The raw materials were 6-bromoquinoline-2-carboxamide from step 3 of Example 55 and 2-(4-chlorophenoxy)-N-((1r,4r)-4-(hydrazine carbonyl)cyclohexyl)acetamide from step 2 of Example 53.

[0412] 6-Bromoquinoline-2-carboxamide (105 mg, 322.29 μmol) and 2-(4-chlorophenoxy)-N-((1r,4r)-4-(hydrazine carbonyl)cyclohexyl)acetamide (78 mg, 322.29 μmol) were added to a sealed tube, dissolved in N,N-dimethylformamide, and reacted at 130 °C for 5 h. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give 17 mg of a white solid, with a yield of 9.75%. 1 H NMR (300MHz, DMSO-d6) δ14.35(d,J=169.1Hz,1H),8.48(dd,J=30.8,8.6Hz,1H),8.36(d,J=20.6Hz,1 H),8.29–8.18(m,1H),8.07(d,J=9.3Hz,1H),8.02(s,1H),7.94(dd,J=16.1,8.9Hz,1H),7.35(dd,J= 7.7,5.4Hz,2H),7.09–6.91(m,2H),4.47(d,J=11.2Hz,2H),3.71(s,1H),2.95–2.65(m,1H),2.11(d, J=12.7Hz,2H),1.90(d,J=12.0Hz,2H),1.56(dq,J=61.1,12.9,12.1Hz,4H).MS(ESI)m / z:calculated for C 25 H 23 BrClN5O2[M+H] +:541.85,found:542.07..

[0413] Example 57

[0414] Synthesis of (5-(4-chloro-3-fluorophenyl)-1,3,4-oxadiazol-2-yl)(4-(5-(5-chlorobenzofuran-2-yl)-1,3,44-oxadiazol-1-yl)piperidin-1-yl)methyl ketone (II-13)

[0415]

[0416] Following the method described in step 3 of Example 1, 72 mg of a white solid was obtained, with a yield of 87.2%. 1 H NMR(400MHz, DMSO-d6)δ7.91(d,J=2.3Hz,1H),7.90–7.87(m,1H),7.85(d,J=8. 8Hz,1H),7.82–7.70(m,3H),7.55(dd,J=8.8,2.2Hz,1H),4.26(d,J=13.3Hz,1H) ,4.09(d,J=13.6Hz,1H),3.59–3.39(m,2H),3.11(t,J=11.4Hz,1H),2.27–2.13( m,2H),1.93(t,J=10.9Hz,1H),1.78(q,J=10.1Hz,1H).MS(ESI)m / z:calculated for C 24 H 16 Cl2FN5O4, [M+H]+: 527.06, found: 527.35.

[0417] Example 58

[0418] Synthesis of (4-(5-(5-chloro-1H-indol-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)(5-(4-chloro-3-fluorophenyl)-1,3,4,4-oxadiazol-1-yl)methyl ketone (II-14)

[0419]

[0420] The raw materials are 2-(5-chloro-1H-indol-2-yl)-5-(piperidin-4-yl)-1,3,4-oxadiazole from step 2 of Example 47 and 5-(4-chloro-3-fluorophenyl)-1,3,4-oxadiazole-2-carboxylic acid from step 1 of Example 10.

[0421] Following the method described in step 3 of Example 1, 27 mg of a white solid was obtained, with a yield of 72.17%. 1H NMR(400MHz,DMSO-d6)δ12.46(d,J=2.1Hz,1H),8.08(dd,J=9.5,1.9Hz,1H),7.98 –7.86(m,2H),7.75(d,J=2.0Hz,1H),7.49(d,J=8.7Hz,1H),7.27(dd,J=8.8,2.0H z,1H),7.22–7.16(m,1H),4.48(dd,J=40.9,13.6Hz,2H),3.69–3.47(m,2H),2.31 –2.15(m,2H),2.04–1.82(m,2H),0.86(t,J=6.7Hz,1H).MS(ESI)m / z:calculated for C 24 H 17 Cl2FN6O3[M+H]+:527.07,found:527.31.

[0422] Example 59

[0423] Synthesis of intermediate 5-chlorophenylfuran-2-carboxamide

[0424]

[0425] Step 1: Synthesis of 5-chlorophenylfuran-2-carboxamide

[0426] Following the method described in step 1 of Example 55, 2.12 g of white solid was obtained, with a yield of 87.24%. 1 H NMR (300MHz, Chloroform-d) δ9.12(s,2H),7.69(dd,J=1.9,0.9Hz,1H),7.38(dt,J=8.8,0.8Hz,1H),7.29(dd,J=8.8,2.0Hz,1H),7.18(dd,J=2.1,0.9Hz,1H).

[0427] Step 2: Synthesis of 5-chlorophenylfuran-2-onitrile

[0428] Following the method described in step 2 of Example 55, 779 mg of a white solid was obtained, with a yield of 77.41%. 1 H NMR (300MHz, DMSO-d6) δ8.09(d,J=1.0Hz,1H),7.96(dd,J=2.2,0.6Hz,1H),7.81(dt,J=9.0,0.8Hz,1H),7.64(dd,J=8.9,2.2Hz,1H).

[0429] Step 3: Synthesis of 5-chlorophenylfuran-2-carboxamide

[0430] Following the method described in step 3 of Example 55, 257 mg of white solid was obtained, with a yield of 98.73%. 1 H NMR (300MHz, DMSO-d6) δ8.30(s,1H),8.25(s,1H),8.17–8.12(m,2H),8.07(d,J=2.2Hz,1H),7.78(d,J=8.9Hz,1H),7.62(dd,J=8.9,2.2Hz,1H).

[0431] Example 60

[0432] Synthesis of N-((1r,4r)-4-(5-(5-chlorobenzofuran-2-yl)-4H-1,2,4-triazol-3-yl)cyclohexyl)-2-(4-chlorophenoxy)acetamide (II-15)

[0433]

[0434] Referring to Example 56, 21 mg of a white solid was obtained, with a yield of 30.20%. 1 H NMR (300MHz, DMSO-d6) δ14.18 (s, 1H), 8.07 (d, J = 8.1 Hz, 1H), 7.78 (d, J = 2. 2Hz,1H),7.71(d,J=8.8Hz,1H),7.43–7.31(m,3H),7.06–6.94(m,2H),4.47 (s,2H),2.88–2.72(m,1H),2.07(d,J=12.4Hz,2H),1.93–1.80(m,2H),1.61(dt,J=13.7,11.0Hz,2H),1.52–1.35(m,2H).MS(ESI)m / z:calculatedfor C 24 H 22 Cl2N4O3[M+H]+:485.11,found:485.11..

[0435] Example 61

[0436] Synthesis of (4-(5-(5-chloro-1H-indol-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)(3-(4-chlorophenyl)-1,2,4-oxadiazol-5-yl)methyl ketone (II-16)

[0437]

[0438] The raw materials are 2-(5-chloro-1H-indol-2-yl)-5-(piperidin-4-yl)-1,3,4-oxadiazole from step 2 of Example 47 and 3-(4-chlorophenyl)-1,2,4-oxadiazole-5-carboxylic acid from step 3 of Example 47.

[0439] Following the method described in step 3 of Example 1, 22 mg of a white solid was obtained, with a yield of 69.84%. 1 H NMR (400MHz, DMSO-d6) δ12.67–12.34(m,1H),8.14–8.04(m,2H),7.75(d,J=2.1Hz,1H),7. 73–7.66(m,2H),7.49(d,J=8.7Hz,1H),7.26(dt,J=8.6,2.3Hz,1H),7.18(dd,J=2.2,0.9Hz ,1H),4.42(d,J=13.3Hz,1H),4.22(d,J=13.6Hz,1H),3.56(ddt,J=14.7,10.7,5.4Hz,2H) ,2.37–2.16(m,2H),1.98–1.83(m,2H),0.86(t,J=6.2Hz,1H).MS(ESI)m / z:calculatedfor C 24 H 18 Cl2N6O3[M+H]+:509.08,found:509.53..

[0440] Example 62

[0441] Synthesis of intermediate 2-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)-2-oxoacetylhydrazine

[0442]

[0443] Step 1: Synthesis of ethyl 2-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)-2-oxoethyl acetate

[0444] The raw materials were 2-(5-chlorobenzofuran-2-yl)-5-(piperidin-4-yl)-1,3,4-oxadiazole and oxaloyl chloride monoethyl ester, obtained in step 3 of Example 39. Following the method described in step 2 of Example 3, 217 mg of a white solid was obtained, with a yield of 92.7%. 1H NMR (300MHz, Chloroform-d) δ7.70 (dd, J=2.1, 0.6Hz, 1H), 7.57 (dt, J=8.8, 0.7Hz, 1H), 7. 47(d,J=0.9Hz,1H),7.43(dd,J=8.9,2.1Hz,1H),4.48–4.32(m,3H),3.85(dtd,J=13.9,4. 4,1.3Hz,1H),3.38(dddd,J=13.8,10.4,5.4,3.6Hz,2H),3.18(dddd,J=13.9,10.7,3.3Hz, 1H),2.36–2.20(m,2H),2.06(ddtd,J=24.3,14.4,10.5,4.1Hz,2H),1.40(t,J=7.1Hz,3H).

[0445] Step 2: Synthesis of 2-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)-2-oxoacetylhydrazine.

[0446] Following the method described in step 1 of Example 3, the reaction was carried out at 95°C. 154 mg of a white solid was obtained, with a yield of 72.17%. MS (ESI) m / z: calculated for C 17 H 16 ClN5O4[M+H]+:390.09,found:390.21.

[0447] Example 63

[0448] Synthesis of intermediate (1s,3s)-3-(trifluoromethoxy)cyclobutane-1-carboxylic acid

[0449]

[0450] Step 1: Synthesis of benzyl(1s,3s)-3-hydroxycyclobutane-1-carboxylate

[0451] Benzyl 3-oxocyclobutane-1-carboxylate (436 mg, 2.13 mmol) was dissolved in 10 mL of anhydrous methanol. NaBH4 (85 mg, 2.24 mmol) was added sequentially at -30 °C, and the reaction was allowed to proceed for 3 h. A saturated solution of NH4Cl was added, and the filtrate was concentrated to remove volatiles. The solution was extracted with EA, and the organic phase was collected and concentrated to give 407 mg of a colorless, transparent liquid, with a yield of 92.33%. 1H NMR (300MHz, Chloroform-d) δ7.45–7.33(m,5H),4.27–4.14(m,1H),2.75–2.55(m,3H),2.48–2.32(s,1H),2.26–2.15(m,2H).

[0452] Step 2: Synthesis of benzyl(1s,3s)-3-(trifluoromethoxy)cyclobutane-1-carboxylate

[0453] In a three-necked flask wrapped in aluminum foil and cooled to -35°C, AgOTf(I) (7.40 g, 28.80 mmol), Selectfluor (5.10 g, 14.40 mmol), KF (2.23 g, 38.40 mmol), and benzyl (1s, 3s)-3-hydroxycyclobutane-1-carboxylate (1.98 g, 9.60 mmol) dissolved in anhydrous ethyl acetate were added sequentially. The mixture was allowed to stand for 30 min under N2 protection. Then, 2-fluoropyridine (2.54 ml, 28.80 mmol) and TMSCF3 (4.26 ml, 28.80 mmol) were added dropwise to maintain the internal temperature below -30°C. The mixture was then transferred to room temperature and reacted overnight. The reaction solution was filtered through diatomaceous earth, washed with EA, concentrated, and the residue eluted by silica gel column chromatography (PE / EA = 200 / 1) to give 1.30 g of a colorless, transparent liquid, with a yield of 49.38%. 1 H NMR(300MHz,Chloroform-d)δ7.38(d,J=14.1Hz,5H),5.17(d,J=2.1Hz,2H),4.59(t,J=7.6Hz,1H),2.85–2.48(m,5H).19F NMR(282MHz,Chloroform-d)δ-59.55.MS(ESI)m / z:calculated forC 13 H 13 F3O3[M+Na]+:297.24,found:297.11

[0454] Step 3: Synthesis of (1s,3s)-3-(trifluoromethoxy)cyclobutane-1-carboxylic acid

[0455] Benzyl(1S,3S)-3-(trifluoromethoxy)cyclobutane-1-carboxylate (1.21 g, 4.40 mmol) was dissolved in THF, and the mixture was reacted overnight with NaOH (880 mg, 22 mmol). The reaction mixture was also concentrated, and the residue was extracted with dichloromethane / 1 mol HCl aqueous solution. The organic phase was collected and evaporated to dryness to give 793 mg of a colorless liquid, yielding 97.94%. 1H NMR (400MHz, DMSO-d6) δ12.63 (s, 1H), 4.47 (p, J = 7.1Hz, 1H), 3.95 (s, 2H), 3.81 3.70(m,1H),2.75(tdt,J=9.0,5.7,2.4Hz,2H),2.15-2.03(m,2H).

[0456] Example 64

[0457] Synthesis of (4-(5-(5-(chlorofuran-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)(5-((1s,3s)-3-(trifluoromethoxy)cyclobutyl)-1,3,4-oxadiazol-1-yl)methyl ketone (II-17)

[0458]

[0459] Step 1: Synthesis of (1s, 3s)-N'-(2-(4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)-2-oxoacetyl)-3-(trifluoromethoxy)cyclobutane-1-formylhydrazine

[0460] Following the method described in step 3 of Example 1, 157 mg of a white solid was obtained, with a yield of 73.21%. 1 H NMR (300MHz, DMSO-d6) δ10.16(s,2H),7.91(d,J=2.2Hz,1H),7.84(d,J=8.9Hz, 1H),7.78(s,1H),7.54(dd,J=8.9,2.2Hz,1H),4.81(p,J=7.6Hz,1H),4.23(d,J= 13.0Hz,2H),4.13–3.95(m,2H),3.06(t,J=12.0Hz,2H),2.78–2.69(m,1H),2.3 0(q,J=9.6Hz,2H),2.15(d,J=13.1Hz,2H),1.83(dq,J=25.7,11.7,10.7Hz,3H).

[0461] Step 2: Synthesis of (4-(5-(5-(chlorofuran-2-yl)-1,3,4-oxadiazol-2-yl)piperidin-1-yl)(5-((1s,3s)-3-(trifluoromethoxy)cyclobutyl)-1,3,4-oxadiazol-1-yl)methyl ketone

[0462] Following the synthesis in step 3 of Example 39, 25 mg of a white solid was obtained, with a yield of 27.13%. 1H NMR(400MHz,Chloroform-d)δ7.70(d,J=2.1Hz,1H),7.57(d,J=8.8Hz,1H),7.48(s,1H),7.43(dd,J=8.8,2.1Hz, 1H),5.00–4.91(m,1H),4.76(p,J=7.6Hz,1H),4.62(dt,J=13.8,4.4Hz,1H),3.71(ddd,J=13.9,10.7,3.1Hz,1H) ,3.44(tdd,J=10.1,7.5,5.2Hz,2H),3.34(ddd,J=13.8,10.7,3.2Hz,1H),2.94(dddt,J=9.9,7.4,5.3,2.6Hz,2H ),2.80(tdd,J=10.3,8.0,2.8Hz,2H),2.34(dt,J=12.6,3.9Hz,2H),2.19–2.06(m,2H).MS(ESI)m / z:calculated forC 23 H 19 ClF3N5O5[M+H]+:528.10,found:528.88..

[0463] Example 65

[0464] Synthesis of intermediate 5-chloro-N-(1-(2-hydrazino-2-oxoacetyl)piperidin-4-yl)benzofuran-2-carboxamide

[0465]

[0466] Step 1: Synthesis of 5-chloro-N-(piperidin-4-yl)benzofuran-2-carboxamide

[0467] The starting material was 5-chlorophenylfuran-2-carboxylic acid and tert-butyl 4-aminopiperidine-1-carboxylate obtained in step 2 of Example 1. The synthesis was performed according to step 3 of Example 1. After debonding with 1.5 ml of trifluoroacetic acid, 2.3 g of a white ester solid was obtained, with a yield of 84.31%. 1HNMR (300MHz, DMSO-d6) δ8.92(d,J=7.6Hz,1H),8.68(s,1H),7.89(d,J=2.2Hz,1H),7.71(d,J=8.8Hz,1H),7.58(d,J=1.0Hz,1H),7.49(dd,J=8.8, 2.2Hz,1H),4.15(d,J=5.2Hz,1H),4.12–4.02(m,1H),3.32(s,1H),3.03( td,J=12.6,3.1Hz,2H),1.97(dd,J=13.8,3.9Hz,2H),1.86–1.72(m,2H).

[0468] Step 2: Synthesis of ethyl 2-(4-(5-chlorophenylfuran-2-carboxamide)piperidin-1-yl)-2-oxoethyl acetate

[0469] Following the method described in step 2 of Example 3, 1.74 g of white solid was obtained, with a yield of 72.3%. 1 H NMR(400MHz,Chloroform-d)δ7.65(d,J=2.0Hz,1H),7.44(d,J=8.6Hz,2H),7.38(dd,J=8.8,2.1Hz ,1H),6.69(d,J=8.1Hz,1H),4.54(ddt,J=13.5,4.5,2.6Hz,1H),4.35(q,J=7.1Hz,2H),4.28(ddt, J=11.7,8.2,4.0Hz,1H),3.75(ddt,J=13.8,5.1,2.8Hz,1H),3.28(ddd,J=14.1,11.9,2.8Hz,1H), 2.93(ddd,J=13.5,12.0,3.0Hz,1H),2.27–2.09(m,2H),1.71–1.51(m,2H),1.37(t,J=7.1Hz,3H).

[0470] Step 3: Synthesis of 5-chloro-N-(1-(2-hydrazino-2-oxoacetyl)piperidin-4-yl)benzofuran-2-carboxamide

[0471] Following the method described in step 1 of Example 3, the reaction was carried out at 85°C. 543 mg of a white solid was obtained, with a yield of 60.17%. 1HNMR(400MHz,DMSO-d6)δ9.77(s,1H),8.75(d,J=7.9Hz,1H),7.88(d,J=2.3Hz,1H), 7.70(d,J=8.8Hz,1H),7.55(s,1H),7.49(dd,J=9.0,2.3Hz,1H),4.51(d,J=23.5Hz, 1H),4.43(s,1H),4.27(d,J=13.3Hz,1H),4.17–4.02(m,1H),3.79(d,J=13.5Hz,1H) ,3.21–3.13(m,1H),2.90–2.78(m,1H),1.85(d,J=11.9Hz,2H),1.63–1.42(m,2H).

[0472] Example 66

[0473] Synthesis of 5-chloro-N-(1-(5-((1s,3s)-3-(trifluoromethoxy)cyclobutyl)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)benzofuran-2-carboxamide (I-29)

[0474]

[0475] Step 1: Synthesis of 5-chloro-N-(1-(2-oxo-2-(2-((1s,3s)-3-(trifluoromethoxy)cyclobutane-1-carbonyl)hydrazyl)acetyl)piperidin-4-yl)benzofuran-2-carboxamide

[0476] The starting materials were 5-chloro-N-(1-(2-hydrazino-2-oxoacetyl)piperidin-4-yl)benzofuran-2-carboxamide obtained in step 3 of Example 65, and (1s,3s)-3-(trifluoromethoxy)cyclobutane-1-carboxylic acid obtained in step 3 of Example 63. Following the synthesis described in step 3 of Example 1, 203 mg of a white solid was obtained, with a yield of 54.89%. 1H NMR(400MHz, DMSO-d6)δ10.05(s,1H),8.79(t,J=7.5Hz,2H),7.93–7.89(m,1H),7.70(s,1H ),7.55–7.51(m,1H),7.48(d,J=2.3Hz,1H),4.82(p,J=7.6Hz,1H),4.31(d,J=14.7Hz,1H), 4.12(dd,J=7.4,3.8Hz,1H),3.98(d,J=13.3Hz,1H),2.98–2.81(m,2H),2.72(tt,J=9.4,7. 5Hz,1H),2.60–2.52(m,2H),2.36–2.19(m,2H),1.57(dddt,J=37.1,19.3,11.4,6.0Hz,4H).

[0477] Step 2: Synthesis of 5-chloro-N-(1-(5-((1s,3s)-3-(trifluoromethoxy)cyclobutyl)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)benzofuran-2-carboxamide

[0478] Following the synthesis in step 3 of Example 39, 26.3 mg of a white solid was obtained, with a yield of 21.07%. 1 H NMR(400MHz,Chloroform-d)δ7.66(d,J=2.0Hz,1H),7.42(s,2H),7.38(dd,J=9.0,2.1Hz,1 H),6.63(d,J=8.1Hz,1H),4.98(dq,J=14.1,3.3Hz,1H),4.80–4.68(m,2H),4.35(tdt,J=11 .6,8.2,4.2Hz,1H),3.53–3.36(m,2H),3.16–3.02(m,1H),2.91(dddt,J=9.9,7.4,5.3,2.7 Hz,2H),2.84–2.70(m,2H),2.32–2.17(m,2H),1.79–1.57(m,2H).MS(ESI)m / z:calculated forC 22 H 20 ClF3N4O5[M+H]+:513.87,found:513.81..

[0479] Example 67

[0480] Synthesis of intermediate 5-chloro-N-(1-(2-hydrazino-2-oxoacetyl)piperidin-4-yl)-1H-indole-2-carboxamide

[0481]

[0482] Step 1: Synthesis of 5-chloro-N-(piperidin-4-yl)-1H-indole-2-carboxamide

[0483] The starting materials were 5-chloro-1H-indole-2-carboxylic acid and tert-butyl 4-aminopiperidine-1-carboxylate. Following the synthesis described in step 1 of Example 65, 1.56 g of a white solid was obtained. The yield was 72.14%. 1 H NMR (400MHz, DMSO-d6) δ11.82(d,J=2.2Hz,1H),8.77(s,1H),8.59(d,J=7.6Hz,1H),7.71(d,J=2.0Hz,1H),7.44(d,J=8.7Hz,1H),7 .23–7.16(m,2H),4.09(dtt,J=10.3,7.0,3.4Hz,1H),3.38–3.31(m,2H),3.12–2.97(m,2H),2.06–1.95(m,2H),1.82–1.66(m,2H).

[0484] Step 2: Synthesis of ethyl 2-(4-(5-chloro-1H-indol-2-carboxamide)piperidin-1-yl)-2-oxoethyl acetate

[0485] Following the method described in step 2 of Example 3, 1.23 g of white solid was obtained, with a yield of 43.21%. 1 H NMR (400MHz, DMSO-d6) δ11.81(d,J=2.2Hz,1H),8.45(d,J=8.0Hz,1H),7.71(d,J=2.0Hz,1H),7.43(d ,J=8.7Hz,1H),7.18(dd,J=8.7,2.1Hz,1H),7.14(dd,J=2.1,0.9Hz,1H),4.31(qd,J=7.1,1.4Hz,2H) ,4.27–4.20(m,1H),4.15(ddq,J=11.5,8.0,4.1,3.7Hz,1H),3.62–3.51(m,1H),3.33–3.23(m,1H),2 .94(td,J=12.8,2.9Hz,1H),1.92(dd,J=13.2,3.9Hz,2H),1.60–1.38(m,2H),1.28(t,J=7.1Hz,3H).

[0486] Step 3: Synthesis of 5-chloro-N-(1-(2-hydrazino-2-oxoacetyl)piperidin-4-yl)-1H-indole-2-carboxamide

[0487] Following the synthesis in step 3 of Example 65, 431 mg of a white solid was obtained, with a yield of 46.98%.1 H NMR (400MHz, DMSO-d6) δ11.85–11.75(m,1H),9.79(s,1H),8.46(d,J=7.6Hz,1H),7.70(d,J=2.1Hz ,1H),7.43(d,J=8.7Hz,1H),7.18(dd,J=8.8,2.1Hz,1H),7.16(d,J=1.6Hz,1H),4.50(d,J=47.4Hz ,2H),4.27(d,J=13.2Hz,1H),4.18–4.03(m,1H),3.78(d,J=13.9Hz,1H),3.19(ddd,J=14.1,11.9, 2.7Hz,1H),2.87(td,J=12.7,2.9Hz,1H),1.97–1.81(m,2H),1.50(dqd,J=39.8,12.4,4.5Hz,2H).

[0488] Example 68

[0489] Synthesis of 5-chloro-N-(1-(5-((1s,3s)-3-(trifluoromethoxy)cyclobutyl)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)-1H-indole-2-carboxamide (I-30)

[0490]

[0491] Step 1: Synthesis of 5-chloro-N-(1-(2-oxo-2-(2-((1s,3s)-3-(trifluoromethoxy)cyclobutane-1-carbonyl)hydrazyl)acetyl)piperidin-4-yl)-1H-indole-2-carboxamide

[0492] The starting materials were 5-chloro-N-(1-(2-hydrazino-2-oxoacetyl)piperidin-4-yl)-1H-indole-2-carboxamide obtained in step 3 of Example 67, and (1s,3s)-3-(trifluoromethoxy)cyclobutane-1-carboxylic acid obtained in step 3 of Example 63. Following the synthesis described in step 3 of Example 1, 925 mg of a white solid was obtained, with a yield of 41.11%. 1H NMR (400MHz, DMSO-d6) δ11.80(d,J=2.1Hz,1H),10.58(s,1H),10.04(s,1H),8.45(d,J=7.8Hz,1H),7.71(d,J=2.0Hz,1H ),7.43(d,J=8.7Hz,1H),7.18(dd,J=8.9,2.0Hz,2H),4.82(p,J=7.6Hz,1H),4.27(d,J=13.2Hz,1H),4.14(pt,J=7.2,3. 6Hz,1H),3.98(d,J=13.4Hz,1H),3.31–3.20(m,1H),2.91(td,J=12.9,3.0Hz,1H),2.72(tt,J=9.4,7.5Hz,1H),2.60–2. 51(m,2H),2.36–2.21(m,2H),1.91(dt,J=13.2,7.3Hz,2H),1.58(qd,J=12.1,4.2Hz,1H),1.46(qd,J=12.3,4.3Hz,1H).

[0493] Step 2: Synthesis of 5-chloro-N-(1-(5-((1s,3s)-3-(trifluoromethoxy)cyclobutyl)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)-1H-indole-2-carboxamide (I-30)

[0494] Following the synthesis in step 2 of Example 66, 57 mg of a white solid was obtained, with a yield of 58.67%. 1 H NMR (400MHz, DMSO-d6) δ8.45(d,J=7.8Hz,1H),7.71(d,J=2.0Hz,1H),7.43(d,J=8.7Hz,1H),7.18(dd,J=8.9,2.0H z,2H),4.82(p,J=7.6Hz,1H),4.27(d,J=13.2Hz,1H),4.14(pt,J=7.2,3.6Hz,1H),3.98(d,J=13.4Hz,1H),3.31–3 .20(m,1H),2.91(td,J=12.9,3.0Hz,1H),2.72(tt,J=9.4,7.5Hz,1H),2.60–2.51(m,2H),2.36–2.21(m,2H),1.91 (dt,J=13.2,7.3Hz,2H),1.58(qd,J=12.1,4.2Hz,1H),1.46(qd,J=12.3,4.3Hz,1H).MS(ESI)m / z:calculatedfor C 22 H 21ClF3N5O4[M+H]+:511.89,found:511.74..

[0495] Example 69

[0496] Synthesis of intermediate 2-((1s,3s)-3-(trifluoromethoxy)cyclobutoxy)acetic acid

[0497]

[0498] Step 1: Synthesis of (1s,3s)-3-(benzyloxy)cyclobutane-1-ol

[0499] Following the synthesis in step 1 of Example 63, 5.40 g of a colorless liquid was obtained, with a yield of 91.37%. 1 H NMR(500MHz,Chloroform-d)δ7.42–7.34(m,2H),7.32–7.22(m,3H),4.60(t,J=1.0H z,2H),3.99(d,J=4.9Hz,1H),3.88(s,1H),3.66(d,J=4.9Hz,1H),1.98–1.85(m,3H).

[0500] Step 2: Synthesis of 2-(3-p-(benzyloxy)cyclobutoxy)tert-butyl acetate

[0501] The starting materials were 3-cis-(benzyloxy)cyclobutanol and tert-butyl 2-chloroacetate obtained in step 1 of Example 63. 3-cis-(benzyloxy)cyclobutanol (1.30 g, 7.29 mmol) and CsCO3 (2.81 g, 14.59 mmol) were dissolved in DMF (10 ml), and after 10 min, tert-butyl 2-chloroacetate (1.10 g, 7.29 mmol) was added, and the reaction was carried out overnight. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give 2.10 g of a colorless liquid, with a yield of 98.47%. 1 H NMR (400MHz, Chloroform-d) δ7.38-7.27(m,5H),4.42(s,2H),3.89(s,2H),3.75-3.62(m,2H),2.65(m,1H),2.09-1.99(s,4H),1.47(s,9H).

[0502] Step 3: Synthesis of 2-(3-cis-hydroxycyclobutoxy)tert-butyl acetate

[0503] Under nitrogen protection, Pd / C (230 mg, 10% Pd / C) was added to 25 ml of MeOH containing 2 g (684 mmol) of 2-(3-p-(benzyloxy)cyclobutoxy)acetic acid tert-butyl ester. The mixture was stirred at 50 °C for 5 h under nitrogen protection. The reaction mixture was filtered through diatomaceous earth, and the filtrate was evaporated to dryness to give 1.27 g of a pale yellow liquid, yielding 91.80%. 1 H NMR (400MHz, Chloroform-d) δ3.95-3.89(m,1H),3.89(s,2H),3.71-1.62(m,1H),2.77-2.67(m,2H),2.00-1.91(m,2H),1.80(s,1H),1.47(s,9H).

[0504] Step 4: Synthesis of 2-(3-chloro-(trifluoromethoxy)cyclobutoxy)tert-butyl acetate

[0505] Following the synthesis in step 2 of Example 63, 1.01 g of a colorless liquid was obtained, with a yield of 41.22%. 1 H NMR (300MHz, Chloroform-d) δ4.36-4.26(m,1H),3.89(s,2H),3.82-3.73(m,1H),2.86-2.70(m,2H),2.37-2.28(m,2H),1.47(s,9H).

[0506] Step 5: Synthesis of 2-((1s,3s)-3-(trifluoromethoxy)cyclobutoxy)acetic acid

[0507] 1.01 g of 2-(3-chloro-(trifluoromethoxy)cyclobutoxy)acetic acid tert-butyl ester was dissolved in 8 ml of DCM, and 2 ml of TFA was added. The reaction mixture was allowed to react overnight. The reaction solution was concentrated and used directly in the next step without purification. 800 mg of a pale yellow liquid was obtained, with a yield of 100%. 1 NMR (400MHz, Chloroform-d): δ4.26-4.19(m,1H),4.00(s,2H),3.73-3.70(m,1H),2.77-2.74(m,2H),2.27-2.24(m,2H).

[0508] Example 70

[0509] Synthesis of N-((1r,4R)-4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexyl)-2-(((1s,3S)-3-(trifluoromethoxy)cyclobutoxy)acetamide (II-18)

[0510]

[0511] The starting materials were (1r,4r)-4-(5-(5-chlorobenzofuran-2-yl)-1,3,4-oxadiazol-2-yl)cyclohexane-1-amine obtained in step 2 of Example 40, and 2-((1s,3s)-3-(trifluoromethoxy)cyclobutoxy)acetic acid obtained in step 5 of Example 69. Following the synthesis in step 3 of Example 1, 34 mg of a white solid was obtained, with a yield of 41.22%. 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.07 (d, J = 8.2 Hz, 1H), 7.90 (s, 1H), 7.84 (d, J = 8.9 Hz, 1H), 7. 77(d,J=6.9Hz,1H),7.54(d,J=9.0Hz,1H),4.49(s,2H),4.26-4.19(m,1H),3.72(d,J=9.2Hz ,1H),3.72-3.70(m,1H),3.11-2.93(m,3H),2.19(d,J=13.5Hz,2H),2.27-2.24(m,2H),1.91 (d,J=12.3Hz,2H),1.67(d,J=12.4Hz,2H),1.47(q,J=12.6Hz,2H); MS(ESI)m / z:calculated forC 23 H 23 ClF3N3O5[M+H] + :514.90,found:514.56.

[0512] Example 71

[0513] Synthesis of intermediate 6-bromo-N-(1-(2-hydrazino-2-oxoacetyl)piperidin-4-yl)quinoline-3-carboxamide

[0514]

[0515] Step 1: Synthesis of 6-bromo-N-(piperidin-4-yl)quinoline-3-carboxamide

[0516] Following the synthesis procedure of step 1 in Example 65, 1.02 g of a yellowish-white solid was obtained. The yield was 74.15%. 1H NMR(300MHz,Chloroform-d)δ8.34(d,J=8.5Hz,1H),8.23(d,J=8.5Hz,1H),8.16 (d,J=8.5Hz,1H),8.06(d,J=2.2Hz,1H),8.02(d,J=9.0Hz,1H),7.85(dd,J=9.0,2 .2Hz,1H),4.16(dddd,J=15.1,10.9,8.4,4.1Hz,1H),3.23(dt,J=12.7,3.7Hz,2H ),2.85(td,J=11.9,2.6Hz,2H),2.13(dd,J=13.1,3.7Hz,2H),1.75–1.54(m,2H).

[0517] Step 2: Synthesis of ethyl 2-(4-(6-bromoquinoline-3-carboxamide)piperidin-1-yl)-2-oxoethyl acetate

[0518] Following the synthesis in step 2 of Example 65, 1.3 g of a white solid was obtained, with a yield of 87.88%. 1 H NMR(300MHz,Chloroform-d)δ8.33(d,J=8.5Hz,1H),8.29–8.23(m,1H),8.19(d,J=8.4Hz,1H),8 .07(d,J=2.1Hz,1H),8.00(d,J=9.0Hz,1H),7.86(dd,J=9.0,2.2Hz,1H),4.56(dd,J=11.6,6.9H z,1H),4.43–4.25(m,3H),3.86–3.70(m,1H),3.32(ddd,J=14.2,11.8,2.8Hz,1H),3.00(ddd,J= 13.6,11.8,3.0Hz,1H),2.30–2.13(m,2H),1.71(pd,J=11.9,4.3Hz,2H),1.40(t,J=7.1Hz,3H).

[0519] Step 3: Synthesis of 6-bromo-N-(1-(2-hydrazino-2-oxoacetyl)piperidin-4-yl)quinoline-3-carboxamide

[0520] Following the synthesis in step 3 of Example 65, 534 mg of a white solid was obtained, with a yield of 78.06%. 1H NMR (400MHz, DMSO-d6) δ9.43(d,J=274.5Hz,1H),8.86(d,J=8.4Hz,1H),8.54(d,J=8.6Hz,1H),8.41(d,J= 2.2Hz,1H),8.19(d,J=8.5Hz,1H),8.09(d,J=9.0Hz,1H),7.99(dd,J=9.0,2.2Hz,1H),4.62–4.38(m,2H),4 .30(d,J=13.2Hz,1H),4.15(dddd,J=15.8,10.5,8.0,4.7Hz,1H),3.82(d,J=13.6Hz,1H),3.27–3.13(m,1H ), 2.86(td,J=13.0,3.1Hz,1H), 1.90(ddd,J=12.3,8.1,3.9Hz,2H), 1.65(dqd,J=28.6,12.3,4.2Hz,2H).

[0521] Example 72

[0522] Synthesis of 6-bromo-N-(1-(5-((1s,3s)-3-(trifluoromethoxy)cyclobutyl)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)quinoline-3-carboxamide (I-31)

[0523]

[0524] Step 1: Synthesis of 6-bromo-N-(1-(2-oxo-2-(2-((1s,3s)-3-(trifluoromethoxy)cyclobutane-1-carbonyl)hydrazyl)acetyl)piperidin-4-yl)quinoline-3-carboxamide

[0525] Following the synthesis in step 1 of Example 66, 347 mg of a white solid was obtained, with a yield of 53.25%. 1H NMR (400MHz, DMSO-d6) δ10.42(d,J=105.6Hz,1H),10.05(s,1H),8.85(d,J=8.3Hz,1H),8.55(d,J=8.5Hz,1H),8.41(d ,J=2.2Hz,1H),8.19(d,J=8.5Hz,1H),8.09(d,J=9.0Hz,1H),7.99(dd,J=9.0,2.3Hz,1H),4.82(p,J=7.6Hz,1H),4.30( d,J=13.2Hz,1H),4.25–4.09(m,1H),4.02(t,J=10.6Hz,1H),3.27(t,J=12.5Hz,1H),2.90(t,J=12.3Hz,1H),2.72(tt ,J=9.2,7.4Hz,1H),2.60–2.51(m,2H),2.38–2.19(m,2H),1.92(d,J=7.4Hz,2H),1.68(dqd,J=41.3,12.2,4.2Hz,2H).

[0526] Step 2: Synthesis of 6-bromo-N-(1-(5-((1s,3s)-3-(trifluoromethoxy)cyclobutyl)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)quinoline-3-carboxamide

[0527] Following the synthesis in step 2 of Example 66, 40 mg of a white solid was obtained, with a yield of 22.51%. 1 H NMR(400MHz,Chloroform-d)δ8.32(d,J=8.4Hz,1H),8.23(d,J=8.6Hz,1H),8.20(d,J=8.2Hz,1H),8.05 (s,1H),7.97(d,J=8.9Hz,1H),7.84(d,J=9.1Hz,1H),5.08–4.90(m,1H),4.74(p,J=7.0,6.1Hz,2H),4. 37(ddp,J=11.8,8.5,4.2Hz,1H),3.63–3.35(m,2H),3.25–3.05(m,1H),3.02–2.86(m,2H),2.78(q,J=1 0.2Hz,2H),2.26(dt,J=12.9,4.0Hz,2H),1.77(dqd,J=24.6,12.0,4.2Hz,2H).MS(ESI)m / z:calculated for C 23 H 21 BrF3N5O4[M+H] + :569.35,found:569.21.

[0528] Example 73

[0529] Synthesis of intermediate 2-(trifluoromethoxy)ethane-1-ol

[0530]

[0531] Step 1: Synthesis of (2-(trifluoromethoxy)ethoxy)methyl)benzene

[0532] The starting material was 2-benzyloxyethanol. Following step 2 of Example 63, a colorless liquid of 1.37 g was obtained, with a yield of 45.76%. 1 HNMR(300MHz,Chloroform-d)δ7.31(s,5H),4.60(s,2H),4.16-4.10(m,2H),3.74-3.67(m,2H).

[0533] Step 2: Synthesis of 2-(trifluoromethoxy)ethane-1-ol

[0534] The colorless liquid obtained in step 3 of Example 69 was 782 mg, with a yield of 96.63%. 1 H NMR (300MHz, Chloroform-d) δ4.16-4.10 (m, 2H), 3.74 (t, J = 4.5Hz, 2H).

[0535] Example 74

[0536] Synthesis of 6-bromo-N-(1-(5-(2-(trifluoromethoxy)ethoxy)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)quinoline-3-carboxamide (I-32)

[0537]

[0538] Step 1: Synthesis of 2-(trifluoromethoxy)ethyl-2-(2-(4-(6-bromoquinoline-3-carboxamide)piperidin-1-yl)-2-oxoacetyl)hydrazine-1-carboxylic acid ester

[0539] At 0 °C, pyridine (134 mg, 1.69 mmol) was added to 2-(trifluoromethoxy)ethane-1-ol (146 mg, 1.13 mmol) and triphosgene (67 mg, 225.48 μmol) in THF (3 mL). After stirring for 60 min, 6-bromo-N-(1-(2-hydrazino-2-oxoacetyl)piperidin-4-yl)quinoline-3-carboxamide (327 mg, 567.39 μmol) was added, and the reaction was carried out overnight at room temperature. The reaction solution was extracted with EA / H2O, and the EA phase was collected. The EA phase was dried over Na2SO4, filtered, the filtrate was concentrated, and the residue was purified by silica gel column chromatography to give 134 mg of white solid, with a yield of 46.53%. 1 H NMR (400MHz, DMSO-d6) δ10.42(d,J=105.6Hz,1H),10.05(s,1H),8.85(d,J=8.3Hz,1H),8.55(d,J=8 .5Hz,1H),8.41(d,J=2.2Hz,1H),8.19(d,J=8.5Hz,1H),8.09(d,J=9.0Hz,1H),7.99(dd,J=9.0,2.3H z,1H),4.30(d,J=13.2Hz,1H),4.25-4.09(m,3H),4.02(t,J=10.6Hz,1H),4.00-3.96(m,2H),3.27(t ,J=12.5Hz,1H),2.90(t,J=12.3Hz,1H),1.92(d,J=7.4Hz,2H),1.68(dqd,J=41.3,12.2,4.2Hz,2H).

[0540] Step 2: Synthesis of 6-bromo-N-(1-(5-(2-(trifluoromethoxy)ethoxy)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)quinoline-3-carboxamide

[0541] Following the synthesis in step 2 of Example 66, 27 mg of a white solid was obtained, with a yield of 32.11%. 1H NMR(300MHz,Chloroform-d)δ8.33(d,J=8.5Hz,1H),8.29–8.23(m,1H),8.19(d,J=8.4Hz,1H),8.07(d,J=2.1Hz,1H), 8.00(d,J=9.0Hz,1H),7.86(dd,J=9.0,2.2Hz,1H),4.40(s,2H),4.56(dd,J=11.6,6.9Hz,1H),4.43–4.25(m,3H),3.9 8(d,J=12.3Hz,1H),3.86–3.70(m,1H),3.82(d,J=12.5Hz,1H),3.32(ddd,J=14.2,11.8,2.8Hz,1H),3.00(ddd,J=13. 6,11.8,3.0Hz,1H),2.30–2.13(m,2H),1.71(pd,J=11.9,4.3Hz,2H),1.40(t,J=7.1Hz,3H).MS(ESI)m / z:calculated for C 21 H 19 BrF3N5O5[M+H] + :558.31,found:558.75.

[0542] Example 75

[0543] Synthesis of 5-chloro-N-(1-(5-(2-(trifluoromethoxy)ethoxy)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)benzofuran-2-carboxamide (I-33)

[0544]

[0545] Step 1: Synthesis of 2-(trifluoromethoxy)ethyl-2-(2-(4-(5-chlorobenzofuran-2-carboxamide)piperidin-1-yl)-2-oxoacetyl)hydrazine-1-carboxylate

[0546] Following the synthesis in step 1 of Example 74, 178 mg of a white solid was obtained, with a yield of 50.83%. 1H NMR (300MHz, DMSO-d6) δ11.27(s,1H),9.49(s,1H),8.92(d,J=7.6Hz,1H),7.89 (d,J=2.2Hz,1H),7.71(d,J=8.8Hz,1H),7.58(d,J=1.0Hz,1H),7.49(dd,J=8.8, 2.2Hz,1H),4.23(s,2H),4.15(d,J=5.2Hz,1H),4.12–4.02(m,3H),3.32(s,1H) ,3.03(td,J=12.6,3.1Hz,2H),1.97(dd,J=13.8,3.9Hz,2H),1.86–1.72(m,2H).

[0547] Step 2: Synthesis of 5-chloro-N-(1-(5-(2-(trifluoromethoxy)ethoxy)-1,3,4-oxadiazol-2-carbonyl)piperidin-4-yl)benzofuran-2-carboxamide

[0548] Following the synthesis in step 2 of Example 66, 19 mg of a white solid was obtained, with a yield of 22.36%. 1 H NMR(400MHz,Chloroform-d)δ7.66(d,J=2.0Hz,1H),7.42(s,2H),7.38(dd,J=9.0,2.1Hz,1H),6.63(d,J=8.1Hz,1H),4.98(dq,J=14.1,3.3Hz,1 H),4.80–4.68(m,1H),4.40-4.35(m,3H),4.02–3.81(m,2H),3.53–3.36(m,2H),2.32–2.17(m,2H),1.79–1.57(m,2H).MS(ESI)m / z:calculated for C 20 H 18 ClF3N4O6[M+H] + :503.83,found:503.09..

[0549] Example 76

[0550] Synthesis of 5-chloro-N-(1-(3-(4-chlorophenyl)-1,2,4-oxadiazol-5-carbonyl)piperidin-4-yl)-1H-indole-2-carboxamide (I-34)

[0551]

[0552] The starting materials were 3-(4-chlorophenyl)-1,2,4-oxadiazole-5-carboxylic acid obtained in step 3 of Example 11 and 5-chloro-N-(piperidin-4-yl)-1H-indole-2-carboxamide obtained in step 1 of Example 67. Following the synthesis described in step 1 of Example 66, 47 mg of a white solid was obtained, with a yield of 54.71%. 1 H NMR (400MHz, DMSO-d6) δ11.79(d,J=2.1Hz,1H),8.46(d,J=7.8Hz,1H),8.19–7.98(m,2H),7.70(d, J=2.1Hz,2H),7.68(d,J=2.0Hz,1H),7.43(dt,J=8.8,0.7Hz,1H),7.22–7.06(m,2H),5.75(s,1H), 4.46(d,J=13.3Hz,1H),4.27–4.16(m,1H),3.43(td,J=13.9,12.9,2.8Hz,1H),3.26–3.06(m,2H), 2.04(s,1H),1.92(d,J=12.7Hz,1H),1.63(tdd,J=23.9,12.1,4.3Hz,2H).MS(ESI)m / z:calculated forC 23 H 19 Cl2N5O3[M+H] + :485.34,found:485.08..

[0553] Experimental Example 1

[0554] ATF4 inhibition rate and IC 50 Measurement

[0555] HEK-293T cells in logarithmic growth phase were seeded in 12-well plates and cultured adherently for 12 h. After incubation with the target compound (500 nM) for 1 h, they were then incubated with carotenoid (Tg, 100 nM) for 3 h. The original culture medium was discarded, and the cells were washed three times with PBS buffer. Cell lysis buffer was added on ice, and protein samples were collected for SDS-PAGE electrophoresis. The samples were then transferred to an NC membrane. After blocking at room temperature for 1 h, the cells were incubated overnight at 4°C with ATF4 primary antibody solution (1:1000). The cells were washed three times with TBST solution and incubated at room temperature for 1 h with HRP-labeled secondary antibody solution. After washing three times with TBST solution, the cells were exposed using a chemiluminescence analyzer. The grayscale values ​​of the bands were analyzed using ImageJ software.

[0556] Table 1. Inhibition rate and partial IC50 of the compounds against ATF4 in HEK-293T cells. 50 value

[0557]

[0558]

[0559] As shown in the table, the compounds in this invention significantly inhibited ATF4 expression in HEK-293T cells. Compared to the lead compound ISRIB, compounds I-13, I-14, I-18, I-19, II-2, and II-4 showed slightly better or roughly equivalent inhibitory activities. Compared to the lead compound ISRIB, compounds I-14, I-18, and II-2 showed significantly higher ATF4 IC50 values. 50 The results showed good numerical values, indicating excellent activity. This suggests that the compound has the ability to activate eIF2B, reverse nerve damage caused by ISR, and treat neurodegenerative diseases.

[0560] Experimental Example 2

[0561] Protein translation rate measured by SDS-PAGE

[0562] CHO cells in the logarithmic growth phase were seeded into 6-well plates and cultured adherently for 12 h. The original culture medium was discarded, the cells were washed once with PBS solution, and serum-free culture medium containing the test compound (500 nM) was added for 1 h. Then, the cells were incubated with carotenoid (Tg, 100 nM) for 24 h. After 24 hours of treatment, cell supernatant was collected and PMSF was added. The mixture was centrifuged at 2000×g and 4℃ for 10 min to remove cell debris. 400 μL of methanol was added and mixed vigorously. 200 μL of chloroform was added, and the mixture was mixed vigorously and vortexed. The mixture was centrifuged at 14000×g for 10 min at 4℃ and the supernatant was discarded. 400 μL of methanol was added to the precipitate and mixed. The mixture was centrifuged at 17000×g for 10 min at 4℃ and the supernatant was discarded. The precipitate was dried and loaded with sample buffer. This was the protein sample in the supernatant. SDS-PAGE electrophoresis was performed, followed by Coomassie brilliant blue staining. The sample was then washed with destaining solution until the background was clean and exposed using a chemiluminescence apparatus.

[0563] Table 2. Determination of the effects of compounds on protein translation in CHO cells.

[0564]

[0565]

[0566] As shown in the table, the compounds in this invention have a good promoting effect on protein translation in CHO cells. Compared with the lead compound ISRIB, compounds I-2, I-9, I-10, I-11, I-15, I-16, I-17, I-25, I-26, I-28, and II-5 showed better protein translation recovery rates and were close to or exceeded the values ​​of the blank group. This indicates that the compounds can significantly promote the recovery of protein synthesis by activating eIF2B, and can reverse nerve damage caused by ISR, thus treating neurodegenerative diseases.

Claims

1. A compound as shown below, 2. A pharmaceutical composition, characterized by, The compound of claim 1 as an active ingredient or a main active ingredient, supplemented with a pharmaceutically acceptable carrier.

3. Use of the compound of claim 1 in the preparation of a medicament for treating and / or preventing neurodegenerative diseases.

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