Nitrogen-containing heterocyclic compound and application thereof

Abstract

The invention discloses a nitrogen-containing heterocyclic compound and application thereof. The nitrogen-containing heterocyclic compound has a structure as shown in a formula (I). The compound provided by the invention has an excellent glutamine cyclase inhibition effect, can be applied to treatment of tumors, immune diseases, neurological diseases or aging, and has a good application prospect.

Description

Technical Field

[0001] This invention relates to the field of chemical and pharmaceutical technology, to a nitrogen-containing heterocyclic compound and its application, and particularly to a nitrogen-containing heterocyclic compound, its application in inhibiting glutamine cyclase, a pharmaceutical composition comprising the nitrogen-containing heterocyclic compound, and its application. Background Technology

[0002] Cancer is a leading cause of death worldwide, and combating cancer is a serious challenge to human health. For many years, chemotherapy, radiotherapy, surgery, and immunotherapy have been common medical treatments for cancer. Cancer immunotherapy is a treatment method that enhances the body's natural immune defenses to fight tumors. In recent years, the rapid development of this method has brought new hope to the treatment of cancer. Existing cancer immunotherapy drugs are mainly antibody drugs targeting immune checkpoints such as PD-1 / PD-L1 and CTLA-4 / B7. However, these drugs suffer from problems in clinical practice, including immune-related toxicity, poor pharmacokinetic properties, weak penetration into tumor cells, and a narrow range of applicable populations. Meanwhile, tumor cells can achieve immune escape by inducing immunosuppression and reducing their own immunogenicity. For example, by highly expressing immunosuppressive signaling molecules that interact with immune cells, such as the interaction between PD-L1 and PD1, or CD47 and SIRPα, they express "don't eat me" signals, directly inhibiting the immune response and promoting tumor cell evasion of immune surveillance.

[0003] Glutaminyl cyclase (EC2.3.2.5) is an enzyme that catalyzes the intramolecular cyclization of N-terminal glutamine residues in peptides and proteins to generate pyroglutamic acid (pGlu*). It has important biological functions such as altering the N-terminal chemical structure of proteins, regulating activity, and enhancing stability.

[0004] Pyroglutamylation is a post-translational modification in which glutamine or glutamate amino acids are converted to the pyroglutamic acid moiety. Currently, two genes in the human body are known to encode enzymes involved in catalyzing this N-terminal pyroglutamylation reaction: glutamine acyl cyclase / protein (QPCT) encoded by the glutamine-peptide cyclotransferase gene (QPCT) and glutamine acyl cyclase-like protein (QPCTL) encoded by the glutamine-peptide cyclotransferase-like gene (QPCTL).

[0005] QPCT catalyzes the intramolecular cyclization of N-terminal glutamine residues to pyroglutamic acid (pGlu*), releasing ammonia. The gene encoding QPCT is located on chromosome 2p22.3. Human QPCT consists of 361 amino acids and has an N-terminal secretion signal. QPCT was first isolated by Messer in 1963 from the latex of the tropical plant Carica papaya (Messer, M. 1963, Nature 4874, 1299). Twenty-four years later, the corresponding enzymatic activity was found in the animal pituitary gland (Busby, WHJ et al. 1987, J Biol Chem 262, 8532-8536; Fischer, WH and Spiess, J. 1987, Proc Natl Acad Sci USA 84, 3628-3632). For mammalian QPCT, the conversion of Gln to pGlu via QPCT can be confirmed by precursors of TRH and GnRH (Busby, WHJ et al. 1987, J Biol Chem 262, 8532-8536; Fischer, WH and Spiess, J. 1987, Proc Natl Acad Sci USA 84, 3628-3632). Furthermore, initial localization experiments of QPCT showed co-localization with its putative catalytic product in the bovine pituitary gland, further enhancing its proposed function in peptide hormone synthesis (Bockers, TM et al. 1995, J Neuroendocrinol 7, 445-453). It has been confirmed that the enzymatic activity of recombinant human QPCT and QPCT derived from brain extracts catalyzes the cyclization of N-terminal glutamine and glutamate. Most surprisingly, it was found that approximately pH 6.0 favors the cyclase-catalyzed Glu-conversion, while the conversion of Gln- to pGlu-derived compounds occurs at an optimal pH of approximately 8.0. Because inhibiting the activity of recombinant human QPCT and QPCT-derived from porcine pituitary extract can inhibit the formation of pGlu-Aβ-related peptide, the enzyme QPCT is a target for drug development for the treatment of Alzheimer's disease (J Med. Chem. 2017, 60, 2573-2590; Alzheimers ResTher 2018, 10, 107).

[0006] Glutamine acyl-peptide cyclotransferase-like protein (QPCTL) also catalyzes the intramolecular cyclization of N-terminal glutamine residues to pyroglutamate (pGlu*), releasing ammonia. QPCTL is located in the Golgi complex and is encoded by chromosome 19q13.32. Mammalian QPCTLs were identified in 2008, including human and mouse QPCTL (J. Mol. Biol. 2008, 379, 966-80). Compared to QPCT, QPCTL exhibits 2 to 15-fold lower catalytic activity for several synthetic substrates. Furthermore, QPCT is expressed at higher levels in neuronal tissues, while QPCTL expression shows no significant difference between different tissues and organs (FEBS J. 2009, 276, 6522-36). In nine different mouse strains, the highest enzymatic QPCT / QPCTL activity was observed in the ventral brain, followed by the cortex and hippocampus. QPCT knockout significantly reduced QPCT activity in the mouse brain, particularly in the hypothalamus and plasma, although activity was still detectable in peripheral organs such as the liver and spleen, which may be a result of QPCTL expression (Int. J.Dev. Neurosci. 2014, 36, 64-73). The QPCTL protein contains 382 amino acid residues, including an active catalytic domain (Ser53-Leu382), a transmembrane domain (Leu35-Trp52), and an extracellular domain (Met1-Arg34) (J Biol Chem. 286, 14199-14208).

[0007] CD47 (Cluster of Differentiation 47), also known as integrin-associated protein, belongs to the immunoglobulin superfamily and is expressed on the surface of various cells, including normal cells (such as erythrocytes) and various tumor cells. CD47 binds to the signal regulatory protein SIRPα, thereby mediating a series of cascade reactions such as apoptosis, proliferation, and immune responses. SIRPα is mainly expressed in monocytes, most macrophages in tissue subsets, and granulocytes. Its expression on macrophages is relatively stable and is not affected by factors such as inflammation. After SIRPα binds to the N-terminal domain of CD47, tyrosine residues on the immunoreceptor tyrosine-based inhibitory motif (ITIM) of SIRPα are phosphorylated, recruiting and activating phosphatases SHP-1 / SHP-2, which in turn triggers the dephosphorylation of downstream pathway molecules, releasing a "don't eat me" signal and inhibiting the phagocytic activity of macrophages. The interaction between CD47 and SIRPα ultimately triggers a series of negative cellular regulatory effects, including the inhibition of phagocytosis by macrophages and neutrophils, as well as cytotoxic effects, thereby enabling tumor cells to evade immune surveillance.

[0008] Studies have found that blocking the CD47 / SIRPα interaction can promote the phagocytosis of tumor cells by macrophages, thereby exerting an anti-tumor effect. Clinical data also show that CD47 expression is closely related to the survival rate of cancer patients; patients with high CD47 expression often have poor survival rates and prognoses. Currently, several CD47-targeting antibody drugs have entered clinical trials. In solid tumors and hematological malignancies, CD47 antibodies can effectively inhibit tumor growth; however, these drugs can cause serious adverse reactions. This is because CD47 is also expressed in normal human erythrocytes. CD47 antibodies can bind not only to CD47 on the surface of tumor cells but also to CD47 on the surface of erythrocytes, causing severe erythrocyte toxicity and other side effects. Furthermore, due to the broad expression range of CD47, antibody drugs exhibit antigen silencing effects, requiring high doses or frequent administration to achieve effective treatment. Therefore, there is an urgent clinical need to find new CD47 inhibition strategies to provide effective means for tumor immunotherapy.

[0009] Studies have shown that QPCTLs can specifically regulate the CD47 / SIRPα signaling axis in various cell types, and this regulation depends on QPCTL enzyme activity (Nat. Med. 2019, 25, 612-619; Cell Res. 2019, 29, 502-505). Furthermore, knocking out QPCTLs in cells or using QPCTL chemical inhibitors effectively inhibits the N-terminal pyroglutamate level of CD47 and the binding of CD47 / SIRPα, but has no significant effect on the expression of CD47 on the cell surface. QPCTLs regulate the binding of CD47 and SIRPα by catalyzing the formation of pyroglutamate from glutamine at position 19 of CD47. In addition, increased phagocytosis of opsonized cells in vitro and increased clearance of opsonized tumor cells in vivo have been shown to be effects of blocking QPCTL activity or production (Nat. Med. 2019, 25, 612-619). In QPCTL knockout mice, the absence of QPCTL promotes the development of macrophages in the bone marrow (Cell Res. 2019, 29, 502-505) and enhances neutrophil-mediated tumor cell killing (Nat. Med. 2019, 25, 612-619; Cancer Sci 2021, 112, 3029-3040). QPCTL is a Golgi-localized protein that is almost not expressed in mature erythrocytes. Therefore, inhibiting QPCTL protein function to block the CD47 / SIRPα pathway may reduce the impact on CD47 function in normal erythrocytes, thereby avoiding the serious side effects caused by CD47 antibody drugs. In conclusion, QPCTL is a potential target for tumor immunotherapy mediated by the CD47 / SIRPα pathway.

[0010] Besides CD47, QPCT and QPCTL can also pyroglutamate other proteins. For example, β-amyloid protein, known to be involved in Alzheimer's disease, is pyroglutamate-modified by QPCT (Pharmacological Research, 2019 147, 104342), while CC motif chemokine ligand 2 (CCL2) protein is pyroglutamate-modified by QPCTL (EMBO Mol Med, 2011, 3, 510-512). Furthermore, the N-terminus of CX3CL1 can also be pyroglutamate-modified by QPCT and QPCTL (Bioscience Reports, 2017 37, BSR20170712). Recent studies have found that QPCTL can also pyroglutamate the N-terminus of BTN (butyrophilin) ​​family proteins, particularly BTN2A1, BTN3A1, BTN3A2, and BTN3A3 (Cell Mol Immunol, 2024, 21, 362–373). However, due to the significant overlap in enzyme properties and substrate preferences, it cannot be ruled out that QPCT and QPCTL have a certain degree of functional overlap in the pyroglutamylation of these targets.

[0011] However, there is very little research on QPCT or QPCTL inhibitors, and no targeted drugs have been marketed to date.

[0012] Therefore, developing a new QPCT or QPCTL inhibitor to provide new treatments for tumors, immune diseases, or neurological diseases is of great practical significance. Summary of the Invention

[0013] This invention provides a novel QPCT or QPCTL inhibitor for cancer treatment, specifically a nitrogen-containing heterocyclic compound, its use in inhibiting glutamine cyclase, particularly QPCTL protein, a pharmaceutical composition comprising the nitrogen-containing heterocyclic compound, and its use.

[0014] To achieve the above objectives, the present invention provides the following technical solution:

[0015] This invention provides a nitrogen-containing heterocyclic compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof:

[0016]

[0017] in:

[0018] ---- indicates a single key or that the key does not exist;

[0019] m is any integer between 0 and 3, and n is any integer between 0 and 3;

[0020] R1 is a 5-10 heteroaryl group optionally substituted with one or more R7s;

[0021] R2 is cyano, nitro, -C(=O)N(R8)2, -NR8C(=O)R9, -S(=O)2R9 or -NR8S(=O)2R9;

[0022] R3 is a hydrogen atom or a halogen atom;

[0023] R4 is arbitrarily assigned to one or more Cs 1-20 Alkyl-substituted 5-10-membered heteroaryl groups;

[0024] R5 represents a hydrogen atom, a halogen atom, a hydroxyl group, or a carbon atom. 1-20 Alkyl or C 1-20 Alkoxy; optionally, when R5 is C 1-20 In the case of alkyl groups, any carbon atom of R5 is connected to any ring atom of R4 to form a 5-10 membered ring structure;

[0025] R6 represents a hydrogen atom or -NR8C(=O)R9;

[0026] Each R7 is independently a halogen atom, arbitrarily C-treated. 3-10 Cycloalkyl-substituted C 1-20 Alkyl, optionally C 3-10 Cycloalkyl-substituted C 1-20 Alkoxy or optionally C 1-20 Alkyl-substituted C 3-10 Cycloalkyloxy;

[0027] Each R8 atom is independently either a hydrogen atom or a carbon atom. 1-20 alkyl;

[0028] Each R9 atom is independently composed of hydrogen and carbon atoms. 1-20 Alkyl, C 1-20 Halogenated alkyl groups, -NHC 1-20 Alkyl, -C 1-20 Alkylene-C 1-20 Alkoxy, C 3-10 Cycloalkyl, 5-10 membered heterocycloalkyl, 5-10 membered heteroaryl or C 6-10 Aryl; the C 3-10 Cycloalkyl, 5-10 membered heterocycloalkyl, 5-10 membered heteroaryl or C 6-10 Each aryl group is independently and optionally bonded by one or more halogen atoms, C 1-20 Alkyl or -C(=O)OC 1-20 Alkyl substitution.

[0029] In some preferred embodiments, in formula (I):

[0030] m is 0 or 1, n is 0 or 1;

[0031] R1 is a 5-10 nitrogen-containing heteroaryl group optionally substituted with one or more R7s;

[0032] R2 is cyano, nitro, -C(=O)N(R8)2, -NR8C(=O)R9, -S(=O)2R9 or -NR8S(=O)2R9;

[0033] R3 is a hydrogen atom or a halogen atom;

[0034] R4 is arbitrarily assigned to one or more Cs 1-10 Alkyl-substituted 5-10 nitrogen-containing heteroaryl groups;

[0035] R5 represents a hydrogen atom, a halogen atom, a hydroxyl group, or a carbon atom. 1-10 Alkyl or C 1-10 Alkoxy; optionally, when R5 is C 1-10 In the case of alkyl groups, any carbon atom of R5 is connected to any ring atom of R4 to form a 5-6 membered ring structure;

[0036] R6 represents a hydrogen atom or -NR8C(=O)R9;

[0037] Each R7 is independently a halogen atom, arbitrarily C-treated. 3-6 Cycloalkyl-substituted C 1-10 Alkyl, optionally C 3-6 Cycloalkyl-substituted C 1-10 Alkoxy or optionally C 1-10 Alkyl-substituted C 3-6 Cycloalkyloxy;

[0038] Each R8 atom is independently either a hydrogen atom or a carbon atom. 1-10 alkyl;

[0039] Each R9 atom is independently composed of hydrogen and carbon atoms. 1-10 Alkyl, C 1-10 Halogenated alkyl groups, -NHC 1-10 Alkyl, -C 1-10 Alkylene-C 1-10 Alkoxy, C 3-6 Cycloalkyl, 5-6 membered heterocycloalkyl, 5-10 membered heteroaryl or C 6-10 Aryl; the C 3-6 Cycloalkyl, 5-6 membered heterocycloalkyl, 5-10 membered heteroaryl or C 6-10 Each aryl group is independently and optionally bonded by one or more halogen atoms, C 1-10 Alkyl or -C(=O)OC 1-10 Alkyl substitution.

[0040] In some preferred embodiments, in formula (I):

[0041] m is 0 or 1, n is 0 or 1, and m and n are not both 0 at the same time;

[0042] R1 is a pyridinyl group optionally substituted with one or more R7 groups;

[0043] R2 is cyano, nitro, -C(=O)N(R8)2, -NR8C(=O)R9, -S(=O)2R9 or -NR8S(=O)2R9;

[0044] R3 is a hydrogen atom or a halogen atom;

[0045] R4 is arbitrarily assigned to one or more Cs 1-10 Alkyl-substituted groups include: in, Indicates the combination of keys;

[0046] R5 is a hydrogen atom, a halogen atom, a hydroxyl group, or a carbon atom. 1-10 Alkyl; optionally, when R5 is C 1-10 In the case of alkyl groups, any carbon atom of R5 is connected to any ring atom of R4 to form a 5-6 membered ring structure;

[0047] R6 represents a hydrogen atom or -NR8C(=O)R9;

[0048] Each R7 is an independent halogen atom;

[0049] Each R8 atom is independently either a hydrogen atom or a carbon atom. 1-10 alkyl;

[0050] Each R9 atom is independently composed of hydrogen and carbon atoms. 1-10 Alkyl, C 1-10 Halogenated alkyl groups, -NHC 1-10 Alkyl, -C 1-10 Alkylene-C 1-10 Alkoxy, C 3-6 Cycloalkyl, 5-6 membered heterocycloalkyl, 5-10 membered heteroaryl or C 6-10 Aryl; the C 3-6 Cycloalkyl, 5-6 membered heterocycloalkyl, 5-10 membered heteroaryl or C 6-10 Each aryl group is independently and optionally bonded by one or more halogen atoms, C 1-10 Alkyl or -C(=O)OC 1-10 Alkyl substitution.

[0051] In some embodiments, the compound is as shown in formula (II):

[0052]

[0053] Wherein, m, n, R1, R2, R3, R4, R5 and R6 have the definitions described in equation (I).

[0054] In some embodiments, the compound is as shown in formula (III):

[0055]

[0056] Wherein, m, n, R1, R2, R3 and R6 have the definitions described in equation (I);

[0057] p is any integer from 0 to 3, preferably 1 or 2.

[0058] In other embodiments, the compound is as shown in formula (IV):

[0059]

[0060] Wherein, m, n, R1, R2, R3, R4, R5 and R6 have the definitions described in equation (I).

[0061] In some preferred embodiments, the compound is as shown in formula (V):

[0062]

[0063] Wherein, m, n, R2, R4, R5, R8 and R9 have the definitions described in equation (I);

[0064] Preferably, R4 is arbitrarily determined by one or two Cs. 1-10 Alkyl-substituted groups include: in, Indicates the combination of keys;

[0065] Preferably, R5 is a hydrogen atom or a halogen atom.

[0066] In some specific embodiments, the specific structural formula of the nitrogen-containing heterocyclic compound of the present invention is as follows:

[0067]

[0068]

[0069]

[0070]

[0071] The present invention also provides a pharmaceutical composition in which the above-mentioned nitrogen-containing heterocyclic compound or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug is used as the active ingredient. In some embodiments, the pharmaceutical composition contains at least one pharmaceutically acceptable carrier. Those skilled in the art can select the pharmaceutical composition according to actual needs to ensure that the active ingredient therein exerts its full effect.

[0072] This invention also provides a drug combination formulation comprising the above-described nitrogen-containing heterocyclic compound or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug, or the above-described pharmaceutical composition, and an antibody. In some embodiments, the antibody is a PD-1 / PD-L1 antibody or a CD47 antibody.

[0073] The present invention also provides the application of the above-mentioned nitrogen-containing heterocyclic compounds or their pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, isotope-labeled substances or prodrugs, or the above-mentioned pharmaceutical compositions, or the above-mentioned drug combinations, in the inhibition of glutamine cyclase. The above-mentioned nitrogen-containing heterocyclic compounds or their pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, isotope-labeled substances or prodrugs, or the above-mentioned pharmaceutical compositions, or the above-mentioned drug combinations, can act as glutamine cyclase inhibitors, capable of inhibiting glutamine cyclotransferase-like (QPCTL) enzymes and / or glutamine acyltransferase (QPCT), that is, capable of inhibiting, reducing or blocking the activity of QPCTL or QPCT.

[0074] In addition, the present invention also provides the use of the above-mentioned nitrogen-containing heterocyclic compounds or their pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, isotope labels or prodrugs, or the above-mentioned pharmaceutical compositions, or the above-mentioned drug combinations in the preparation of medicaments for the prevention and / or treatment of tumors, immune diseases, neurological diseases or aging.

[0075] In some embodiments, the tumor is selected from at least one of colorectal cancer, lung cancer, gastric cancer, melanoma, myeloma, breast cancer, adenocarcinoma, bladder cancer, and hematologic malignancy.

[0076] In some embodiments, the immune disease is selected from at least one of eczema, alopecia areata, psoriasis, vitiligo, rheumatoid arthritis, lupus syndrome, acne, and hidradenitis suppurativa.

[0077] In some embodiments, the neurological disease is selected from at least one of Alzheimer's disease, Huntington's disease, neurodegeneration of Down syndrome, depression, anxiety disorder, psychosis, and multiple sclerosis.

[0078] The above technical solution is only one feasible technical solution of the present invention. The scope of protection of the present invention is not limited thereto. Those skilled in the art can reasonably adjust the specific design according to actual needs.

[0079] The QPCTL / QPCT activity mentioned above refers to the cyclization of the N-terminal glutamine residue into pyroglutamic acid (pGlu*) or the intramolecular cyclization of the N-terminal L-high glutamine or L-β-high glutamine into cyclopyroglutamine derivatives when ammonia is released.

[0080] EC includes QPCT and QPCTL as the activity of glutamate cyclase (EC), which is further defined as EC activity.

[0081] QPCT inhibitors and glutamine acyl cyclase inhibitors are well known to those skilled in the art, specifically enzyme inhibitors that inhibit the catalytic activity of glutamine acyl cyclase (QPCT) or the activity of its glutamine cyclase (EC).

[0082] QPCTL inhibitors and glutamine cyclotransferase-like protein inhibitors are well known to those skilled in the art, specifically enzyme inhibitors that inhibit the catalytic activity of glutamine cyclotransferase-like protein (QPCTL) or its glutamine cyclase (EC) activity.

[0083] Glutamine cyclase inhibitors include QPCT inhibitors and / or QPCTL inhibitors, which inhibit QPCT / QPCTL and have the same or similar pharmacological effects.

[0084] The effectiveness of QPCTL suppression: In view of the relationship with QPCTL suppression, in the preferred embodiment, the IC of QPCTL suppression is... 50 The concentration is 10 μM or less, preferably 1 μM or less, even more preferably 0.1 μM or less, or 0.01 μM or less, or most preferably 0.001 μM or less. Therefore, although the active substance is described herein as a QPCTL inhibitor for convenience, it should be understood that such nomenclature is not intended to limit the subject matter of the invention to a specific mechanism of action.

[0085] The present invention has the following advantages or beneficial effects:

[0086] This invention provides a novel nitrogen-containing heterocyclic compound that exhibits excellent inhibitory effects on glutamine cyclases (especially QPCTL enzymes), and has promising applications in the treatment of tumors, immune diseases, neurological diseases, or aging. Detailed Implementation

[0087] The present invention will be further described below with reference to specific embodiments, but these are not intended to limit the invention.

[0088] Example 1

[0089] 3-(6-Fluoropyridin-3-yl)-2-[4-(Thiazolyl-5-yl)piperidin-1-yl]-benzyl-1-carboxylonitrile (Compound 1)

[0090] 3-(6-fluoropyridin-3-yl)-2-(4-(thiazol-5-yl)piperidin-1-yl)benzonitrile

[0091]

[0092] The synthetic route for compound 1 is shown above.

[0093] Step a: Compound MC1 (111 mg, 0.36 mmol, 1.5 eq), compound MC2 (39 mg, 0.24 mmol, 1.0 eq), and Na2CO3 (51 mg, 0.48 mmol, 2.0 eq) were dissolved in a mixed solvent of 1,4-dioxane (2 mL) and H2O (0.2 mL). Under argon protection, Pd(dppf)Cl2 (18 mg, 0.024 mmol, 0.1 eq) was added, and the system was heated to 90 °C and reacted for 16 hours. After the reaction was completed, the mixture was cooled to room temperature, extracted with water and EA, and the organic phase was concentrated under reduced pressure and then purified by silica gel column chromatography to obtain a yellow oily compound MC3 (90 mg, 94.0% yield). 1 H NMR (400MHz, CDCl3) δ8.54 (s, 1H), 7.65 (s, 1H), 5.96 (brs, 1H), 3.99-3.96 (m, 2H), 3.54 (t, J = 5.6Hz, 2H), 2.45-2.41 (m, 2H), 1.38 (s, 9H).

[0094] Step b: Intermediate MC3 (1.47 g, 1.0 eq) was dissolved in MeOH (20 mL), and Pd / C (10%, 200 mg) was added under argon protection. The system was purged with hydrogen three times at room temperature, and the reaction was carried out overnight at 45 °C. After the reaction was completed, the mixture was filtered, and the organic phase was concentrated to obtain compound MC4 (1.2 g). 1 HNMR(400MHz, CDCl3)δ8.62(s,1H),7.58(s,1H),4.16-4.11(m,2H),3.03-2.95(m ,1H),2.79(t,J=13.2Hz,2H),1.95-1.91(m,2H),1.63-1.51(m,2H),1.41(s,9H).

[0095] Step c: After removing the Boc protecting group from intermediate MC4, the crude 5-(piperidin-4-yl)thiazole product (0.20 g, 1.21 mmol, 1.0 eq) was dissolved in DMSO (4 mL). Then, 3-bromo-2-fluorobenzonitrile (MC5) (0.29 g, 1.45 mmol, 1.2 eq) and anhydrous potassium carbonate (0.25 g, 1.82 mmol, 1.5 eq) were added sequentially, and the mixture was heated and stirred at 100 °C for 16 hours. After the reaction was completed by TLC monitoring, dichloromethane and water were added for extraction, followed by extraction again with saturated brine and dichloromethane. The organic phase was collected and purified using a SepaBean machine T200 to finally obtain 0.15 g of intermediate MC6.

[0096] In step d, intermediate MC6 (35 mg, 0.10 mmol, 1.0 eq), sodium carbonate (21 mg, 0.2 mmol, 2.0 eq), and 4-fluoro-3-pyridineboronic acid (34 mg, 0.15 mmol, 1.5 eq) were dissolved in a mixed solvent of 2 mL dioxane and 0.2 mL water. Under nitrogen protection, Pd(dppf)Cl2 (7 mg, 0.01 mmol, 0.1 eq) was added, and the reaction was carried out at 90°C for 10 hours. After cooling to room temperature, the reaction mixture was diluted with water and then extracted with ethyl acetate. The organic layer was concentrated and separated by HPLC to obtain a white solid product.

[0097] The data for compound 1 were obtained as follows: 1 H NMR(400MHz,MeOD-d4)δ9.03(s,1H),8.26(d,J=2.5Hz,1H),8.07-7.92(m,1H),7.83-7.63(m,2H),7.54(dd,J=7.7,1.6Hz,1H),7.31(t,J =7.7Hz,1H),7.20(dd,J=8.5,2.6Hz,1H),3.26-3.20(m,1H),3.18-2.98(m,2H),2.07-1.89(m,2H),1.76-1.49(m,2H),1.35-1.28(m,2H). LRMS(ESI)[M+H] + ,found:365.2.

[0098] Example 2

[0099] 3-(6-Fluoropyridin-3-yl)-2-[4-(1-Methyl-1H-1,2,3-triazol-4-yl)piperidin-1-yl]-benzyl-1-carboxylonitrile (Compound 2)

[0100] 3-(6-fluoropyridin-3-yl)-2-(4-(1-methyl-1H-1,2,3-triazol-4-yl)piperidin-1-yl)benzonitrile

[0101]

[0102] The synthetic route for compound 2 is shown above.

[0103] Step a: Compound MC7 (1.05 g, 5.00 mmol, 1.0 eq), trimethylsilyl azidosilane (636 mg, 5.5 mmol, 1.1 eq), and CuI (48 mg, 0.25 mmol, 0.05 equiv) were dissolved in DMF (9 mL) and MeOH (1 mL) under argon protection. The reaction mixture was heated to 100 °C and stirred for 8 h. After the reaction was complete, EA (50 mL) was added for dilution, followed by washing with water (50 mL) and brine (50 mL). The solution was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography to obtain intermediate MC8 (600 mg, 47.6% yield). 1 H NMR (400MHz, CDCl3) δ7.47(s,1H),4.17-4.15(m,2H),2.97-2.82(m,3H),1.99-1.94(m,2H),1.68-1.56(m,2H),1.44(s,9H).

[0104] Step b: Under argon protection at 0°C, intermediate MC8 (30 mg, 0.12 mmol, 1.0 eq) and potassium carbonate (33 mg, 0.24 mmol, 2.0 eq) were dissolved in DMF (2 mL). Iodomethane (25 mg, 0.18 mmol, 1.5 eq) was slowly added to the system, and the mixture was stirred overnight at room temperature. After the reaction was complete, EA (50 mL) was added for dilution, followed by washing with H2O (50 mL) and saline solution (50 mL). The mixture was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography to obtain intermediate MC9 (22 mg, 70.5% yield). ¹H NMR (600 MHz, CDCl₃) δ 7.33 (s, 1H), 4.13–4.10 (m, 5H), 2.88–2.81 (m, 3H), 1.95–1.91 (m, 2H), 1.65–1.57 (m, 2H), 1.45 (s, 9H).

[0105] Steps c and d: Using intermediate MC9, react as in step c of Example 1 to generate intermediate MC10, and then as in step d of Example 1, to obtain the data for compound 2: 1H NMR (400MHz, MeOD-d4) δ8.24(dd,J=2.1,1.1Hz,1H),8.07-7.95(m,1H),7.69(dd,J=7.8,1.7Hz,1H),7.53(dd,J=7.7,1.7Hz,1H),7.43(s,1H),7.29(t, J=7.7Hz,1H),7.19(dd,J=8.4,2.6Hz,1H),4.09(s,3H),3.26-3.15(m,2H), 3.15-3.02(m,2H),2.83-2.66(m,1H),1.92-1.81(m,2H),1.70-1.53(m,2H). LRMS(ESI)[M+H] + ,found:363.2.

[0106] Example 3

[0107] 2-[4-(1H-1,2,3-triazol-1-yl)piperidin-1-yl]-3-(6-fluoropyridin-3-yl)benzyl-1-carboxylonitrile (Compound 3)

[0108] 2-(4-(1H-1,2,3-triazol-1-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile

[0109]

[0110] The synthetic route for compound 3 is shown above.

[0111] Step a: Under argon protection at 0°C, compound MC12 (500 mg, 7.24 mmol, 1.0 eq) was dissolved in DMF (5 mL), and then NaH (320 mg, 60% mmol, 8.0 mmol, 1.1 eq) was slowly added. The reaction was slowly heated to room temperature and stirred for 2 hours. Then, compound MC11 (2.4 g, 8.69 mmol, 1.2 eq) was added to the system, and the system was heated to 60°C and stirred overnight. After the reaction was completed, EA (50 mL) was added for dilution, followed by washing with H2O (50 mL) and brine (50 mL). The solution was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography to obtain intermediate MC13 (900 mg, 49.3% yield). 1H NMR(400MHz, CDCl3)δ7.49(brs,1H),7.45-7.43(m,1H),4.48-4.39(m,1H),4.00(d,J =13.6Hz,2H),2.80-2.74(m,2H),1.97-1.92(m,2H),1.80-1.69(m,2H),1.23(s,9H).

[0112] Steps b and c: Using intermediate MC13, react as in step c of Example 1 to generate intermediate MC14, and then as in step d of Example 1, to obtain the data for compound 3 as follows: 1 H NMR (400MHz, MeOD-d4) δ8.27(dd,J=2.1,1.2Hz,1H),8.10-7.98(m,2H),7.78(d,J=1.2Hz,1H),7.72(dd,J=7.7,1.6Hz,1H),7.56(dd,J=7.7, 1.7Hz,1H),7.33(t,J=7.7Hz,1H),7.22(dd,J=8.5,2.7Hz,1H),4.71-4.53(m,1H),3.30-3.27(m,2H),3.26-3.08(m,2H),2.17-1.98(m,4H). LRMS(ESI)[M+H] + ,found:349.2.

[0113] Example 4

[0114] 2-[4-(1H-1,2,4-triazol-1-yl)piperidin-1-yl]-3-(6-fluoropyridin-3-yl)benzyl-1-carboxylonitrile (Compound 4)

[0115] 2-(4-(1H-1,2,4-triazol-1-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile

[0116]

[0117] The synthesis method is the same as in Example 3, and the route shown above yielded the following data for compound 4: 1H NMR (400MHz, MeOD-d4) δ8.95(d,J=15.8Hz,1H),8.03(td,J=8.0,2.5Hz,1H),7.72(dd,J=7.7,1.7Hz,1H),7.56(dd,J=7.7,1.7Hz,1 H),7.33(t,J=7.7Hz,1H),7.21(dd,J=8.5,2.5Hz,1H),4.56-4.36(m,1H),3.30-3.26(m,2H),3.24-3.06(m,2H),2.14-1.98(m,4H). LRMS(ESI)[M+H] + ,found:349.2.

[0118] Example 5

[0119] 2-[4-(2H-1,2,3-triazol-2-yl)piperidin-1-yl]-3-(6-fluoropyridin-3-yl)benzyl-1-carboxylonitrile (Compound 5)

[0120] 2-(4-(2H-1,2,3-triazol-2-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile

[0121]

[0122] The synthesis method is the same as in Example 3, and the route shown above yielded the following data for compound 5: 1 H NMR (400MHz, DMSO) δ8.28 (d, J=2.5Hz, 1H), 8.05 (td, J=8.2, 2.5Hz, 1H), 7.86-7.73 (m, 3H), 7.56 (dd, J=7.6, 1.7Hz,1H),7.37-7.24(m,2H),4.68-4.48(m,1H),3.26-3.13(m,2H),3.13-2.95(m,2H),2.09-1.84(m,4H). LRMS(ESI)[M+H] + ,found:349.2.

[0123] Example 6

[0124] 2-[4-(4,5-dimethyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl]-3-(6-fluoropyridin-3-yl)benzyl-1-carboxylonitrile (Compound 6)

[0125] 2-(4-(4,5-dimethyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile

[0126]

[0127]

[0128] The synthesis method is the same as in Example 1, and the synthetic route is shown above. The data for compound 6 are as follows: 1 H NMR (400MHz, MeOD-d4) δ8.28(d,J=2.4Hz,1H),8.04(td,J=8.0,2.8Hz,1H),7.75(dd,J=8.0,2.0Hz,1H),7.58(dd,J=7.6,2.0Hz,1H),7.35(t,J=7.6Hz,1 H),7.22(dd,J=8.4,2.8Hz,1H),3.74(s,3H),3.30-3.28(m,2H),3.17-3.07 (m,3H),2.62(s,3H),1.97-1.93(m,2H),1.87-1.77(m,2H).LRMS(ESI)[M+H] + ,found:377.4.

[0129] Example 7

[0130] 3-(6-Fluoropyridin-3-yl)-2-(4-(4-isopropyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzyl-1-carboxylonitrile (Compound 7)

[0131] 3-(6-fluoropyridin-3-yl)-2-(4-(4-isopropyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile

[0132]

[0133] The synthetic route is shown above, and the synthetic method is the same as in Example 1. The data for compound 7 are as follows: 1H NMR (400MHz, MeOD-d4) δ9.47(s,1H),8.56(d,J=2.8Hz,1H),8.52(s,1H),7.82-7.76(m,2H),7.61(dd,J=7.6,1.6Hz,1H),7.37(t, J=8.0Hz,1H),4.82-4.74(m,1H),3.27-3.19(m,4H),1.99-1.94(m,2H),1.89-1.57(m,2H),1.58(d,J=6.8Hz,6H).LRMS(ESI)[M+H] + ,found:391.3.

[0134] Example 8

[0135] 2-(4-(1H-imidazol-5-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzyl-1-carboxylonitrile (Compound 8)

[0136] 2-(4-(1H-imidazol-5-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile

[0137]

[0138] The synthetic route is shown above, and the synthetic method is the same as in Example 1. The data for compound 8 are as follows: 1 H NMR (400MHz, MeOD-d4) δ8.83 (s, 1H), 8.29 (d, J = 2.4Hz, 1H), 8.02 (td, J = 8.0, 2.4Hz,1H),7.74(dd,J=8.0,1.6Hz,1H),7.57(dd,J=7.6,1.6Hz,1H),7.36-7 .32(m,2H),7.21(dd,J=8.4,2.4Hz,1H),3.29-3.24(m,2H),3.16-3.11(m,2H ),2.88-2.79(m,1H),1.98-1.93(m,2H),1.69-1.65(m,2H).LRMS(ESI)[M+H] + ,found:348.2.

[0139] Example 9

[0140] 2-(4-(4-ethyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzyl-1-carboxylonitrile (compound)

[0141] 2-(4-(4-ethyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile

[0142]

[0143] The synthetic route is shown above, and the synthetic method is the same as in Example 1. The data for compound 9 are as follows: 1 H NMR (600MHz, MeOD-d4) δ9.34 (s, 1H), 8.26 (s, 1H), 8.02 (t, J = 7.8Hz, 1H), 7.73 (d ,J=7.8Hz,1H),7.56(d,J=7.8Hz,1H),7.33(t,J=7.8Hz,1H),7.21(dd,J=8.4,2. 4Hz,1H),4.27(q,J=7.2Hz,2H),3.25-3.20(m,2H),3.28(brs,2H),3.21-3.15(m ,1H),1.97-1.93(m,2H),1.85(brs,2H),1.54(t,J=7.2Hz,3H).LRMS(ESI)calcd for C 21 H 22 FN6[M+H] + 377.2, found: 377.3.

[0144] Example 10

[0145] 3-(6-Fluoropyridin-3-yl)-2-(4-(1-methyl-1H-1,2,3-triazol-5-yl)piperidin-1-yl)benzyl-1-carboxynitrile (compound)

[0146] 3-(6-fluoropyridin-3-yl)-2-(4-(1-methyl-1H-1,2,3-triazol-5-yl)piperidin-1-yl)benzonitrile

[0147]

[0148] The synthetic route for compound 10 is shown above:

[0149] Step a: Compound MC32 (251 mg, 1.2 mmol, 1.2 eq), CuSO4·5H2O (3 mg, 0.01 mmol, 0.01 eq), (+)-sodium ascorbate (10 mg, 0.05 mmol, 0.05 eq), and urea (2 mg, 0.02 mmol, 0.02 eq) were dissolved in MeOH / H2O (2 mL / 2 mL). Benzyl azide (133 mg, 1.0 mmol, 1.0 eq) was then added to the system, and the reaction was continued at room temperature for 16 hours. After the reaction was complete, the mixture was diluted with EA (50 mL), washed with H2O (50 mL) and brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography to obtain intermediate MC33 (100 mg, 29.2% yield). 1 H NMR (600MHz, CDCl3) δ7.39-7.32(m,3H),7.27-7.23(m,2H),7.16(s,1H),5.48(s,2H),4 .13-4.09(m,2H),2.93-2.83(m,3H),2.00-1.96(m,2H),1.59-1.51(m,2H),1.44(s,9H).

[0150] Step b: Intermediate MC33 (40 mg, 0.12 mmol, 1.0 eq) was dissolved in MeCN (2 mL), and then methyl iodide (103 mg, 0.72 mmol, 6.0 eq) was added. The reaction system was heated to 60 °C and reacted overnight. After the reaction was completed, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography to obtain intermediate MC34 (50 mg, crude). 1 H NMR(400MHz, CDCl3)δ8.96(s,1H),7.60-7.56(m,2H),7.40-7.37(m,3H),5.90(s,2H),4.38(s, 3H),4.18-4.07(m,2H),3.41-3.32(m,1H),2.92-2.80(m,2H),2.06-1.92(m,4H),1.41(s,9H).

[0151] In step c, intermediate MC34 (50 mg, 1.0 mmol, 1.0 eq) and t-BuOK (29 mg, 2.5 mmol, 2.5 eq) were dissolved in MeCN (10 mL) under argon protection at 0 °C, and the mixture was slowly heated to room temperature and reacted overnight. After the reaction was complete, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography to obtain intermediate MC35 (20 mg, 74.9% yield). 1H NMR(600MHz, CDCl3)δ7.44(s,1H),4.00(s,3H),2.84-2.72(m,3H),1.88(d,J=13.8Hz,2H),1.72(brs,2H),1.63-1.54(m,2H),1.47(s,9H).LRMS(ESI)calcd for C 13 H 23 N4O2[M+H] + 267.2, found:267.3.

[0152] After removing the Boc protecting group using compound MC35 in steps d and e, the synthesis method is the same as step c in Example 1 to obtain intermediate MC36. The data for compound 10 obtained is the same as step d in Example 1. 1 H NMR (400MHz, MeOD-) d4 )δ8.26(d,J=2.4Hz,1H),8.04-7.99(m,1H),7.71(dd,J=8.0,1.6Hz,1H),7.61(s,1H),7.55(dd,J=8.4,1.6Hz,1H),7.31(t,J=7.6Hz ,1H),7.23-7.19(m,1H),4.03(s,3H),3.26-3.14(m,4H),2.92-2.84(m,1H),1.88-1.84(m,2H),1.63-1.59(m,2H).LRMS(ESI)calcd for C 20 H 20 FN6[M+H] + 363.2, found: 363.3.

[0153] Example 11

[0154] 3-(6-Fluoropyridin-3-yl)-2-(4-(3-methyl-4H-1,2,4-triazol-4-yl)piperidin-1-yl)benzyl-1-carboxylonitrile (Compound 11)

[0155] 3-(6-fluoropyridin-3-yl)-2-(4-(3-methyl-4H-1,2,4-triazol-4-yl)piperidin-1-yl)benzonitrile

[0156]

[0157] The synthetic route for compound 11 is shown above:

[0158] Step a: Compound MC37 (814 mg, 11.0 mmol, 1.0 eq) was dissolved in acetonitrile (5 mL), and then DMFDMA (1.35 g, 11.0 mmol, 1.0 eq) was added to the system. The mixture was heated to 50 °C and stirred for 1 hour. Then, compound MC38 (1.0 g, 5.5 mmol, 0.5 eq) and HOAc (1 mL) were added to the system, and the mixture was heated to 100 °C and stirred for 18 hours. After the reaction was completed, the reaction system was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain intermediate MC39 (1.5 g, 52.0% yield). 1 H NMR(600MHz, CDCl3)δ8.00(s,1H),4.19(brs,2H),3.93-3.87(m,1H),2.77-2 .73(m,2H),2.34(s,3H),1.92-1.88(m,2H),1.71-1.63(m,2H),1.35(s,9H).

[0159] Steps b and c: Using intermediate MC39, react as in step c of Example 1 to generate intermediate MC40, and then as in step d of Example 1, synthesize compound 11. The data are as follows: 1 H NMR (600MHz, MeOD-d4) δ9.23(s,1H),8.28(s,1H),8.02(t,J=6.6Hz,1H),7.74(d,J=7.8Hz,1H),7.56(d,J=7.8Hz,1H),7.35(t,J=7.8Hz,1H),7. 21(dd,J=9.0,2.4Hz,1H),4.43-4.37(m,1H),3.33(brs,2H),3.18(br,2H),2.72(s,3H),2.10-2.06(m,2H),1.99-1.96(m,2H).LRMS(ESI)calcd for C 20 H 20 FN6[M+H] + 363.2, found: 363.3.

[0160] Example 12

[0161] 2-(4-(1-ethyl-1H-imidazol-5-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzyl-1-carboxylonitrile (Compound 12)

[0162] 2-(4-(1-ethyl-1H-imidazol-5-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile

[0163]

[0164] The synthetic route is shown above, and the synthetic method is the same as in Example 1. The data for compound 12 obtained are as follows: 1 H NMR (400MHz, MeOD-d4) δ8.87 (s, 1H), 8.28 (d, J = 2.4Hz, 1H), 8.01 (td, J = 8.0, 2.4Hz, 1H), 7. 72(dd,J=7.6,1.6Hz,1H),7.55(dd,J=7.6,1.6Hz,1H),7.36(s,1H),7.32(t,J=7.6Hz,1H), 7.21(dd,J=8.4,2.4Hz,1H),4.23(q,J=7.2Hz,2H),3.27-3.23(m,2H),3.15(brs,2H),2.89 -2.81(m,1H),1.92-1.87(m,2H),1.65-1.61(m,2H),1.53(t,J=7.2Hz,3H).LRMS(ESI)calcd for C 22 H 23 FN5[M+H] + 376.2, found: 376.3.

[0165] Example 13

[0166] 3-(6-Fluoropyridin-3-yl)-2-(4-(4-methylpyridin-3-yl)piperidin-1-yl)benzyl-1-carboxylonitrile (Compound 13)

[0167] 3-(6-fluoropyridin-3-yl)-2-(4-(4-methylpyridin-3-yl)piperidin-1-yl)benzonitrile

[0168]

[0169] The synthetic route is shown above, and the synthetic method is the same as in Example 1. The data for compound 13 are as follows: 1H NMR (400MHz, MeOD-d4) δ8.57-8.53(m,2H),8.31(s,1H),8.04(t,J=8.0Hz,1H),7.88(d,J=6.0Hz,1H),7.73(d,J=8.0Hz,1H),7.57(d,J=8.0Hz, 1H),7.33(t,J=8.0Hz,1H),7.23(dd,J=8.4,2.4Hz,1H),3.29-3.18(m,4H),3.09-3.02(m,1H),2.67(s,3H),1.84-1.70(m,4H).LRMS(ESI)calcd for C 23 H 22 FN4[M+H] + 373.2, found: 373.3.

[0170] Example 14

[0171] 3-(6-Fluoropyridin-3-yl)-2-(4-(3-methylpyridin-4-yl)piperidin-1-yl)benzyl-1-carboxylonitrile (Compound 14)

[0172] 3-(6-fluoropyridin-3-yl)-2-(4-(3-methylpyridin-4-yl)piperidin-1-yl)benzonitrile

[0173]

[0174] The synthetic route is shown above, and the synthetic method is the same as in Example 1. The data for compound 14 are as follows: 1 H NMR (400MHz, MeOD-d4) δ8.64-8.62(m,2H),8.31(s,1H),8.04(t,J=8.0Hz,1H),7.88(d,J=6.0Hz,1H),7.73(d,J=7.6Hz,1H),7.56(d, J=7.6Hz,1H),7.33(t,J=7.6Hz,1H),7.24(dd,J=8.4,2.0Hz,1H),3.30-3.08(m,5H),2.55(s,3H),1.80-1.76(m,4H).LRMS(ESI)calcd for C 23 H 22 FN4[M+H] + 373.2, found: 373.3.

[0175] Example 15

[0176] 3-(6-Fluoropyridin-3-yl)-2-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile (Compound 15)

[0177] 3-(6-fluoropyridin-3-yl)-2-(4-methyl-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile

[0178]

[0179] The synthetic route is shown above, and the synthetic method is the same as in Example 1. The data for compound 15 are as follows: 1 H NMR (600MHz, MeOD-d4) δ9.09 (s, 1H), 8.25 (d, J = 2.4Hz, 1H), 8.00 (td, J = 7.8, 2.4Hz,1H),7.73(dd,J=7.8,1.8Hz,1H),7.58(dd,J=7.8,1.8Hz,1H),7.35(t ,J=7.8Hz,1H),7.19(dd,J=9.0,2.4Hz,1H),3.98(s,3H),3.22(brs,2H),3.1 1(brs,2H),2.28(brs,2H),1.86-1.84(m,2H),1.48(s,3H).LRMS(ESI)[M+H] + ,found:377.2.

[0180] Example 16

[0181] 2-(4-fluoro-4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile (Compound 16)

[0182] 2-(4-Fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile

[0183]

[0184] The synthetic route is shown above, and the synthetic method is the same as in Example 1. The data for compound 16 obtained are as follows: 1H NMR (600MHz, MeOD-d4) δ8.75(s,1H),8.27(d,J=2.4Hz,1H),8.06(td,J=7.8,2.4Hz,1H),7.76(dd,J=7.8,1.2Hz,1H),7.58(dd,J=7.8,1.2Hz ,1H),7.36(t,J=7.8Hz,1H),7.22(dd,J=8.4,2.4Hz,1H),3.90(s,3H),3.40(brs,2H),3.20-3.17(m,2H),2.30-2.23(m,4H).LRMS(ESI)[M+H] + ,found:381.2.

[0185] Example 17

[0186] 3-(6-Fluoropyridin-3-yl)-2-(6-(4-methyl-4H-1,2,4-triazol-3-yl)-3-azabicyclo[3.1.0]hexane-3-yl)benzonitrile (Compound 17)

[0187] 3-(6-Fluoropyridin-3-yl)-2-(6-(4-methyl-4H-1,2,4-triazol-3-yl)-3-azabicyclo[3.1.0]hexan-3-yl)benzonitrile

[0188]

[0189] The synthetic route is shown above, and the synthetic method is the same as in Example 1. The data for compound 17 are as follows: 1 H NMR (600MHz, MeOD-d4) δ8.96(s,1H),8.24(d,J=2.4Hz,1H),7.96(td,J=7.8,2 .4Hz,1H),7.76(dd,J=7.8,1.8Hz,1H),7.59(dd,J=7.8,1.2Hz,1H),7.38(t,J =7.8Hz,1H),7.24(dd,J=8.4,3.6Hz,1H),3.89(s,3H),3.52(d,J=9.6Hz,2H), 3.38(d,J=9.0Hz,2H),2.41-2.39(m,1H),2.27-2.25(m,2H).LRMS(ESI)[M+H] + ,found:361.4.

[0190] Example 18

[0191] 5-Fluoro-3-(6-Fluoropyridin-3-yl)-2-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile (Compound 18)

[0192] 5-fluoro-3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile

[0193]

[0194] The synthetic route is shown above, and the synthetic method is the same as in Example 1. The data for compound 18 are as follows: 1 H NMR (600MHz, MeOD-d4) δ9.10(s,1H),8.43(s,1H),8.20-8.16(m,1H),7.46-7.43(m,1H),7.41-7.39(m,1H),7.25-7.22( m,1H),3.96-3.92(m,5H),3.32-3.28(m,1H),3.06-3.01(m,2H),2.21-2.17(m,2H),2.06-2.01(m,2H).LRMS(ESI)[M+H] + ,found:381.4.

[0195] Example 19

[0196] 3-(6-Fluoropyridin-3-yl)-2-(3-(4-methyl-4H-1,2,4-triazol-3-yl)pyrrolo-1-yl)benzonitrile (Compound 19)

[0197] 3-(6-fluoropyridin-3-yl)-2-(3-(4-methyl-4H-1,2,4-triazol-3-yl)pyrrolidin-1-yl)benzonitrile

[0198]

[0199] The synthetic route is shown above, and the synthetic method is the same as in Example 1. The data for compound 19 are as follows: 1H NMR (400MHz, MeOD-d4) δ9.42 (s, 1H), 8.21 (d, J = 2.4Hz, 1H), 8.01-7.96 (m, 1 H),7.69(dd,J=8.0,1.6Hz,1H),7.35(dd,J=7.6,1.6Hz,1H),7.30(t,J=7.6H z,1H),7.17-7.13(m,1H),3.88(s,3H),3.85-3.78(m,2H),3.60-3.53(m,1H ),3.43-3.30(m,2H),2.49-2.39(m,1H),2.32-2.23(m,1H).LRMS(ESI)calcd for C 19 H 18 FN6[M+H] + 349.2, found:349.3.

[0200] Example 20

[0201] 3-(6-Fluoropyridin-3-yl)-2-(3-(4-methyl-4H-1,2,4-triazol-3-yl)azacyclopropane-1-yl)benzonitrile (Compound 20)

[0202] 3-(6-fluoropyridin-3-yl)-2-(3-(4-methyl-4H-1,2,4-triazol-3-yl)azetidin-1-yl)benzonitrile

[0203]

[0204]

[0205] The synthetic route is shown above, and the synthetic method is the same as in Example 1. The data for compound 20 are as follows: 1 H NMR(400MHz,MeOD-d4)δ9.05(s,1H),8.24(d,J=2.4Hz,1H),8.01-7.95(m,1H),7.58(dd,J=7.6,2.0Hz,1H),7.35(dd,J=7.6,1.6H z,1H),7.15(dd,J=8.4,2.8Hz,1H),6.97(t,J=8.0Hz,1H),4.35(t,J=8.0Hz,2H),4.17-4.07(m,3H),3.70(s,3H).LRMS(ESI)calcd forC 18 H 16 FN6[M+H] + 335.1, found: 335.3.

[0206] Example 21

[0207] 2-(6',7'-dihydro-5'H-spiro[piperidin-4,8'-[1,2,4]triazolo[4,3-a]pyridin]-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile (compound 21)

[0208] 2-(6',7'-dihydro-5'H-spiro[piperidine-4,8'-[1,2,4]triazolo[4,3-a]pyridin]-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile

[0209]

[0210] The synthetic route for compound 21 is shown above:

[0211] Step a: Compound MC60 (536 mg, 2.0 mmol, 1.0 eq) and Lawesson's reagent (409 mg, 2.1 mmol, 1.05 eq) were dissolved in toluene (20 mL). The reaction mixture was heated to reflux and stirred overnight. After the reaction was complete, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography to obtain intermediate MC61 (290 mg, 51.1% yield). 1 H NMR (400MHz, CDCl3) δ8.31 (s, 1H), 4.01 (brs, 2H), 3.34 (brs, 2H), 2.90 (t, J = 11.6H z,2H),2.54(t,J=12.4Hz,2H),1.88(brs,4H),1.52-1.44(m,11H).LRMS(ESI)calcd for C 14 H 25 N₂O₂S[M+H] + 285.2, found:285.3.

[0212] In step b, intermediate MC61 (284 mg, 1.0 mmol, 1.0 eq) and compound MC62 (72 mg, 1.2 mmol, 1.2 eq) were dissolved in DCM (5 mL), followed by the addition of AgOBz (460 mg, 2.0 mmol, 2.0 eq) and HOAc (0.17 mL). The reaction mixture was heated to reflux and stirred overnight. After the reaction was complete, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography to obtain intermediate MC63 (170 mg, 58.0% yield). 1H NMR (400MHz, CDCl3) δ8.08 (s, 1H), 4.01 (t, J = 6.0Hz, 2H), 3.84 (brs, 2H), 3.61 (brs, 2H) ,2.07-1.98(m,4H),1.88-1.85(m,2H),1.63-1.57(m,2H),1.47(s,9H).LRMS(ESI)calcd for C 15 H 25 N4O2[M+H] + 293.2, found:293.3.

[0213] After removing the Boc protecting group using intermediate MC63 in steps c and d, intermediate MC64 is obtained as in step c of the synthetic route in Example 1. Following step d of the synthetic route in Example 1, the data for compound 21 are as follows: 1 H NMR(400MHz,MeOD-d4)δ9.21(s,1H),8.29(d,J=2.4Hz,1H),8.08-8.02(m,1H),7.74(dd,J=8.0,1.6Hz,1H),7.57(dd,J=8.0,2.0Hz,1H),7.3 4(t,J=7.6Hz,1H),7.23(dd,J=8.4,2.4Hz,1H),4.23(t,J=6.4Hz,2H),3.25(brs,4H),2.14-2.00(m,6H),1.81-1.75(m,2H).LRMS(ESI)calcd for C 22 H 22 FN6[M+H] + 389.2, found: 389.3.

[0214] Example 22

[0215] 2-(5',6'-dihydrospiro[piperidine-4,7'-pyrrolo[2,1-c][1,2,4]triazol]-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile (compound 22)

[0216] 2-(5',6'-dihydrospiro[piperidine-4,7'-pyrrolo[2,1-c][1,2,4]triazol]-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile

[0217]

[0218] The synthetic route is shown above, and the synthetic method is the same as in Example 21. The data for compound 22 are as follows: 1 H NMR (400MHz, MeOD-d4) δ9.23 (s, 1H), 8.29 (d, J = 2.4Hz, 1H), 8.06 (td, J = 8.0, 2.4Hz, 1H), 7.75 (dd, J = 7.6, 1.6Hz, 1H), 7.57 (dd, J = 7.6, 1.6Hz, 1H), 7. 34(t,J=7.6Hz,1H),7.24(dd,J=8.4,2.4Hz,1H),4.33(t,J=7.2Hz,2H),3.20-3.14(m,2H),2.72(t,J=7.2Hz,4H),1.94-1.84(m,4H).LRMS(ESI)calcd for C 21 H 20 FN6[M+H] + 375.2, found:375.2.

[0219] Example 23

[0220] 2-(4-(4-ethyl-4H-1,2,4-triazol-3-yl)-4-fluoropiperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile (compound 23)

[0221] 2-(4-(4-ethyl-4H-1,2,4-triazol-3-yl)-4-fluoropiperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile

[0222]

[0223] The synthetic route is shown above, and the synthetic method is the same as in Example 16. The data for compound 23 are as follows: 1 H NMR (600MHz, MeOD-d4) δ9.34(s,1H),8.26(s,1H),8.02(t,J=7.8Hz,1H),7.73(d,J=7.8Hz,1H),7.56(d,J=7.8Hz,1H),7.33(t,J=7.8Hz,1H),7.21 (dd,J=8.4,2.4Hz,1H),4.27(q,J=10.8Hz,2H),3.44-3.34(m,2H),3.18-3.15(m,2H),2.25-2.18(m,4H),1.54(t,J=10.8Hz,3H).LRMS(ESI)calcd for C 21 H 21F2N6[M+H] + 395.2, found: 395.3.

[0224] Example 24

[0225] 3-(6-Fluoropyridin-3-yl)-2-(4-hydroxy-4-(1-methyl-1H-imidazol-5-yl)piperidin-1-yl)benzonitrile (Compound 24)

[0226] 3-(6-fluoropyridin-3-yl)-2-(4-hydroxy-4-(1-methyl-1H-imidazol-5-yl)piperidin-1-yl)benzonitrile

[0227]

[0228] The synthetic route for compound 24 is shown above:

[0229] Step a: Dissolve MC71 (1.32 g, 16.0 mmol, 1.0 eq) in THF (30 mL). Under argon protection, cool the system to 78 °C, slowly add n-BuLi (10 mL, 1.6 M, 1.0 eq), and continue stirring for 1 hour. Then add triethylchlorosilane (2.4 g, 16.0 mmol, 1.0 eq), slowly heat to room temperature, then cool again to -78 °C, slowly add n-BuLi (10 mL, 1.6 M, 1.0 eq), and continue stirring for 1 hour. Then heat the system to -15 °C and cool again to -78 °C. Add a THF (10 mL) solution of MC72 (3.0 g, 15.0 mmol, 0.94 equiv) to the system. Heat the system to room temperature and continue stirring overnight. After the reaction was complete, the system was diluted with EA (50 mL), washed successively with H2O (50 mL) and saline (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The intermediate MC73 (1130 mg, 25.0% yield) was obtained by silica gel column chromatography. 1 HNMR (400MHz, CDCl3): δ7.20(s,1H),6.69(s,1H),3.86(brs,2H),3.79(s,3H),3.27(brs,2H),1.95-1.86(m,4H),1.45(s,9H).

[0230] After removing the Boc protecting group using intermediate MC73 in steps b and c, intermediate MC74 is obtained as in step c of the synthetic route in Example 1, and the data for compound 24 obtained as in step d of the synthetic route in Example 1 are as follows: 1H NMR (400MHz, MeOD-d4) δ8.81(s,1H),8.27(d,J=2.4Hz,1H),8.01(td,J=8.0,2.4Hz,1H),7.72(dd,J=8.0,1.6Hz,1H),7.54(dd,J=7.6,1.6Hz,1H) ,7.43(d,J=7.6Hz,1H),7.31(t,J=7.6Hz,1H),7.20(dd,J=8.4,2.4Hz,1H),3.42(brs,2H),3.06-3.00(m,2H),1.99-1.95(m,4H).LRMS(ESI)calcd for C 21 H 21 FN5O[M+H] + 378.2, found:378.2.

[0231] Example 25

[0232] 3-(6-Fluoropyridin-3-yl)-2-(4-methoxy-4-(1-methyl-1H-imidazol-5-yl)piperidin-1-yl)benzonitrile (Compound 25)

[0233] 3-(6-fluoropyridin-3-yl)-2-(4-methoxy-4-(1-methyl-1H-imidazol-5-yl)piperidin-1-yl)benzonitrile

[0234]

[0235] The synthetic route for compound 25 is shown above:

[0236] Step a: Intermediate MC73 (300 mg, 1.07 mmol, 1.0 eq) was dissolved in DMF (10 mL). NaH (85 mg, 60%, 2.14 mmol, 2.0 eq) was added under argon protection at 0 °C. The system was heated to room temperature and stirred for 2 hours. Then, iodomethane (303 mg, 2.14 mmol, 2.0 eq) was added, and the system was heated to reflux and stirred overnight. After the reaction was complete, the system was cooled to room temperature, diluted with EA (20 mL), washed once with water (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain intermediate MC74 (305 mg, 96.8% yield). 1H NMR(400MHz, CDCl3)δ7.41(s,1H),6.89(s,1H),3.95-3.83(m,2H),3.75(s,3H),3 .23-3.12(m,2H),3.00(s,3H),2.15-2.10(m,2H),1.88-1.80(m,2H),1.47(s,9H).

[0237] After removing the Boc protecting group using intermediate MC74 in steps b and c, intermediate MC75 is obtained as in step c of the synthetic route in Example 1, and the data for compound 25 obtained as in step d of the synthetic route in Example 1 are as follows: 1 H NMR(400MHz,MeOD-d4)δ8.88(s,1H),8.26(d,J=2.0Hz,1H),8.01(td,J=8.0,2.4Hz, 1H),7.72(dd,J=7.6,1.6Hz,1H),7.55(dd,J=8.0,1.6Hz,1H),7.52(d,J=1.6Hz,1H) ,7.32(t,J=8.0Hz,1H),7.19(dd,J=8.4,2.4Hz,1H),4.00(s,3H),3.35(brs,2H),3. 07(s,3H),3.05-3.00(m,2H),2.24-2.19(m,2H),1.94-1.90(m,2H).LRMS(ESI)calcd for C 22 H 23 FN5O[M+H] + 392.2, found: 392.3.

[0238] Example 26

[0239] 3-(6-Fluoropyridin-3-yl)-2-(4-hydroxy-4-(1-methyl-1H-imidazol-4-yl)piperidin-1-yl)benzonitrile (Compound 26)

[0240] 3-(6-fluoropyridin-3-yl)-2-(4-hydroxy-4-(1-methyl-1H-imidazol-4-yl)piperidin-1-yl)benzonitrile

[0241]

[0242] The synthetic route is shown above, and the synthetic method is the same as in Example 24. The data for compound 26 are as follows: 1H NMR(600MHz,MeOD-d4)δ8.28(s,1H),8.03-7.99(m,1H),7.74(dd,J=7.8,1.8Hz,1 H),7.56(dd,J=7.2,1.2Hz,1H),7.50(d,J=1.8Hz,1H),7.44(d,J=2.4Hz,1H),7.3 4(t,J=7.2Hz,1H),7.21(dd,J=8.4,2.4Hz,1H),4.06(s,3H),3.47-3.42(m,2H),3 .07(d,J=12.0Hz,2H),2.13-2.03(m,2H),1.94(d,J=11.4Hz,2H).LRMS(ESI)calcd for C 21 H 21 FN5O[M+H] + 378.2, found: 378.3.

[0243] Example 27

[0244] 3-(6-Fluoropyridin-3-yl)-2-(4-methoxy-4-(1-methyl-1H-imidazol-4-yl)piperidin-1-yl)benzonitrile (Compound 27)

[0245] 3-(6-fluoropyridin-3-yl)-2-(4-methoxy-4-(1-methyl-1H-imidazol-4-yl)piperidin-1-yl)benzonitrile

[0246]

[0247]

[0248] The synthetic route is shown above, and the synthetic method is the same as in Example 25. The data for compound 27 are as follows: 1H NMR(600MHz,MeOD-d4)δ8.26(d,J=2.4Hz,1H),8.04-7.99(m,1H),7.74(dd,J=7.8,1. 8Hz,1H),7.59(d,J=1.8Hz,1H),7.57(dd,J=7.8,1.8Hz,1H),7.53(d,J=2.4Hz,1H),7. 35(t,J=7.2Hz,1H),7.2(dd,J=8.4,2.4Hz,1H),4.04(s,3H),3.43-3.34(m,2H),3.14( s,3H),3.06(d,J=13.8Hz,2H),2.30-2.26(m,2H),2.04-1.98(m,2H).LRMS(ESI)calcd forC 22 H 23 FN5O[M+H] + 392.2, found: 392.3.

[0249] Example 28

[0250] 3-(6-Fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzylamine (Compound 28)

[0251] 3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzamide

[0252]

[0253] The synthetic route of compound 28 is shown above: QP5020 (10 mg, 0.028 mmol, 1.0 eq) and K2CO3 (1.0 mg, 0.0072 mmol, 0.25 eq) were dissolved in DMSO (1.0 mL), and H2O2 (3 μL) was added at room temperature. The reaction system was heated to 50 °C and stirred for 2 h. After the reaction was completed, the system was concentrated under reduced pressure and then purified by prep-HPLC (10% to 100% MeCNin H2O containing 0.1% TFA over 90 min) to obtain compound 28 (2 mg, 20% yield). The data for compound 28 are as follows: 1H NMR (400MHz, MeOD-d4) δ9.13(s,1H),8.30(s,1H),8.08-8.03(m,1H),7.45(d,J=7.6Hz,1H),7.35(d,J=7.6Hz,1H),7.27(t ,J=7.6Hz,1H),7.17(d,J=8.4Hz,1H),3.86(s,3H),3.16-3.03(m,5H),1.87-1.83(m,2H),1.60(brs,2H).LRMS(ESI)calcd forC 20 H 22 FN6O[M+H] + 381.2, found: 381.3.

[0254] Example 29

[0255] 2-(4-fluoro-4-(1-methyl-1H-imidazol-5-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile (Compound 29)

[0256] 2-(4-fluoro-4-(1-methyl-1H-imidazol-5-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile

[0257]

[0258]

[0259] The synthetic route for compound 29 is shown above:

[0260] Step a: Intermediate MC74 (10 mg, 0.027 mmol, 1.0 eq) was dissolved in DCM (2 ml). DAST (18 mg, 0.11 mmol, 4.0 eq) was added to the system at 0 °C, and the mixture was stirred at room temperature for 4 h. After the reaction was complete, the solvent was removed by rotary evaporation under reduced pressure. Intermediate MC80 was obtained as a white oily liquid (7 mg, 71.6% yield) by silica gel column chromatography. 1 HNMR (400MHz, CDCl3) δ7.78 (dd, J=8.0, 1.6Hz, 1H), 7.55 (dd, J=8.0, 1.6Hz, 1H), 7.43 (s, 1H), 7.05-6.99(m,2H),3.85-3.76(m,5H),3.26-3.22(m,2H),2.40-2.28(m,4H).LRMS(ESI)calcd for C 16 H17 BrFN4[M+H] + 363.1, found:363.1.

[0261] Step b, the same as step d in Example 1, yielded the following data for compound 29: 1 H NMR (600MHz, MeOD-d4) δ8.91 (s, 1H), 8.28 (d, J = 2.4Hz, 1H), 8.01 (td, J = 7.8, 2. 4Hz,1H),7.75(dd,J=7.8,2.4Hz,1H),7.65(s,1H),7.57(dd,J=7.2,1.8Hz,1H) ,7.35(t,J=7.8Hz,1H),7.20(dd,J=8.4,2.4Hz,1H),4.01(s,3H),3.43-3.34(m ,2H),3.16-3.13(m,2H),2.29-2.25(m,2H),2.18-2.01(m,2H).LRMS(ESI)calcd for C 21 H 20 F2N5[M+H] + 380.2, found: 380.3.

[0262] Example 30

[0263] N-(2-cyano-4-(6-fluoropyridin-3-yl)-3-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)butyramide (compound 30)

[0264] N-(2-cyano-4-(6-fluoropyridin-3-yl)-3-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)butyramide

[0265]

[0266] The synthesis method of compound 30 is shown above:

[0267] Step a: Compound MC81 (500 mg, 2.325 mmol, 1.0 eq), butyryl chloride (250 mg, 2.325 mmol, 1.0 eq), and DIPEA (460 μL, 1.2 eq) were dissolved in DCM (10 mL) and stirred overnight at room temperature. After the reaction was complete, the mixture was cooled to room temperature, and water and EA were added for extraction. The organic phase was concentrated under reduced pressure and then purified by silica gel column chromatography to obtain a yellow oily compound MC82 (410 mg, 62.4% yield). 1H NMR (400MHz, CDCl3) δ8.22(d,J=9.2Hz,1H),7.73(t,J=8.0Hz,1H),7.64(s,1H),7.23( dd,J=8.4,2.4Hz,1H),2.45(t,J=7.2Hz,2H),1.83-1.73(m,2H),1.03(t,J=7.2Hz,3H).

[0268] Step b: After removing the Cbz protecting group from intermediate MC83, the crude product (0.20 g, 1.21 mmol, 1.0 eq) was dissolved in DMSO (4 mL). Then, intermediate 3 (0.41 g, 1.45 mmol, 1.2 eq) and anhydrous potassium carbonate (0.25 g, 1.82 mmol, 1.5 eq) were added sequentially, and the mixture was heated and stirred at 100 °C for 16 hours. After the reaction was completed by TLC monitoring, dichloromethane and water were added for extraction, followed by extraction again with saturated brine and dichloromethane. The organic phase was collected and purified using a SepaBean machine T200, finally yielding 0.15 g of intermediate 5. 1 H NMR (400MHz, MeOD-d4) δ8.09-8.04(m,2H),7.71(d,J=9.2Hz,1H),7.59(s,1H),3.70(s,3H),3.50-3.43(m,2H),2.96-2.88( m,1H),2.43(t,J=7.2Hz,2H),2.33-2.29(m,2H),1.81-1.75(m,2H),1.64(s,2H),1.31-1.24(m,2H),1.03(t,J=7.2Hz,3H).

[0269] Step c: Intermediate MC84 (43 mg, 0.10 mmol, 1.0 eq), sodium carbonate (21 mg, 0.2 mmol, 2.0 eq), and 4-fluoro-3-pyridineboronic acid (34 mg, 0.15 mmol, 1.5 eq) were dissolved in a mixed solvent of 2 mL dioxane and 0.2 mL water. Under nitrogen protection, Pd(dppf)Cl2 (7 mg, 0.01 mmol, 0.1 eq) was added, and the reaction was carried out at 90°C for 10 hours. After cooling to room temperature, the reaction mixture was diluted with water and then extracted with ethyl acetate. The organic layer was concentrated and separated by HPLC to obtain a white solid product. The data for compound 30 are as follows: 1H NMR (400MHz, MeOD-d4) δ9.09(s,1H),8.27(s,1H),8.03(t,J=8.0Hz,1H),7.54(d,J=8.4Hz,1H),7.41(d,J=8.4Hz,1H),7.23(dd,J=8 .4,2.4Hz,1H),3.88(s,3H),3.16-3.14(m,2H),2.47(t,J=7.2Hz,2H),1.98-1.75(m,6H),1.36-1.31(m,3H),1.08(t,J=7.2Hz,3H). LRMS(ESI)[M+H] + ,found:448.1.

[0270] Example 31

[0271] 2-Fluoro-5-(2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(methanesulfonyl)phenyl)pyridine (compound 31)

[0272] 2-fluoro-5-(2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(methylsulfonyl)phenyl)pyridine

[0273]

[0274] The synthetic route for compound 31 is shown above:

[0275] Step a: 1-Bromo-2-fluorobenzene MC85 (1.75 g, 10.0 mmol, 10.0 eq) was dissolved in tetrahydrofuran (30 mL) under argon protection. After cooling the system to -78 °C, LDA (6 mL, 2 M, 1.2 eq) was added dropwise over 1 h while maintaining the temperature at -78 °C. Then, DMSO (1.1 mL, 13.0 mmol, 1.3 eq) was added to the system, and the reaction was stirred at -78 °C for 2 h. After the reaction was completed, the reaction was terminated with H2O (20 mL), then extracted with EA (50 mL) and H2O (50 mL), washed with brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (PE:EA 20:1–6:1) to obtain compound MC86 (1.4 g, 63.6% yield). 1 H NMR (400MHz, CDCl3) δ7.35 (t, J = 6.8 Hz, 1H), 7.19 (t, J = 8.0 Hz, 1H), 6.98 (t, J = 8.0 Hz, 1H), 3.82 (s, 3H).

[0276] Step b: MC86 (1.40 g, 6.4 mmol, 1.0 equiv) was dissolved in DCM (30 mL) under argon protection. After the reaction system was cooled to 0 °C, m-CPBA (2.2 g, 30.0 mmol, 2.2 eq) was slowly added. The system was slowly warmed to room temperature to continue the reaction. After the reaction was completed, DCM (70 mL) was added to dilute the system, and then the mixture was washed with saturated NaHCO3aq (500 mL) and brine (50 mL). The solution was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography to obtain a white solid compound MC87 (600 mg, 37.4% yield). 1 H NMR (400MHz, CDCl3) δ7.94-7.90(m,1H),7.88-7.83(m,1H),7.27-7.22(m,1H),3.25(s,3H).

[0277] Step c: Using intermediate MC87 and compound MC83, MC88 (105 mg, 21.7% yield) was obtained via the synthetic route step c in Example 1. 1 H NMR (400MHz, CDCl3) δ8.07-8.04(m,2H),7.77(dd,J=8.0,2.4Hz,1H),7.25(t,J=8.0Hz,1H),3.89-3.82(m, 2H),3.68(s,3H),3.58(s,3H),3.16-3.11(m,2H),2.87-2.80(m,1H),2.30-2.18(m,2H),2.03-1.97(m,2H).

[0278] Step d: Using intermediate MC88, as in step d of the synthetic route in Example 1, the data for compound 31 are as follows: 1 HNMR(400MHz,MeOD-d4)δ9.07(s,1H),8.25(d,J=2.4Hz,1H),8.18(dd,J=7.2,2.4Hz,1H),7.98(td,J=8.0,2.4Hz,1H),7.54-7.48(m,2H),7.24(dd ,J=8.4,2.4Hz,1H),3.82(s,3H),3.41-3.34(m,5H),2.95-2.88(m,1H),2.66-2.59(m,2H),2.12-2.01(m,2H),1.92-1.88(m,2H).LRMS(ESI)calcd for C 20 H 23 FN5O2S[M+H] +416.2, found: 416.3.

[0279] Example 32

[0280] 2-Fluoro-5-(2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-nitrobenzene)pyridine (compound 32)

[0281] 2-fluoro-5-(2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-nitrophenyl)pyridine

[0282]

[0283] The synthesis route is shown above:

[0284] Step a: After removing the Cbz protecting group from compound MC83, 4-(4-methyl-4H-[1,2,4]triazol-3-yl)-piperidine (1.0 g, 6.02 mmol, 1.0 eq) was dissolved in DMSO (15 mL). Potassium carbonate (2.5 g, 18.06 mmol, 3.0 eq) and 1-bromo-2-fluoro-3-nitrobenzene (MC89) (2.6 g, 12.04 mmol, 2 eq) were added to the system. The mixture was heated to 60 °C and reacted with stirring for 4 hours. After cooling to room temperature, the mixture was filtered, and the solution was concentrated under pressure. The solution was then washed with DCM and water, and the organic phase was concentrated under reduced pressure. After purification by silica gel column chromatography, 0.65 g of intermediate MC90 (30% yield) was obtained. 1 H NMR(400MHz,MeOD-d4)δ8.40(s,1H),7.91(d,J=7.6Hz,1H),7.66(dd,J=7.6,2.4Hz,1H),7.16(t,J=7.6Hz,1H) ,3.79(s,3H),3.63-3.56(m,2H),3.43-3.32(m,2H),3.17-3.08(m,1H),2.26-2.15(m,2H),2.10-2.05(m,2H).

[0285] Step b: Intermediate 1-(2-bromo-6-nitrobenzene)-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidine (0.65 g, 1.78 mmol, 1.0 eq), Na₂CO₃ (0.38 g, 3.56 mmol, 2.0 eq), and pinacol ester of 2-fluoropyridine 5-borate (0.6 g, 2.67 mmol, 1.5 eq) were dissolved in a mixed solvent of 1,4-dioxane (9 mL) and H₂O (1 mL). After purging the reaction system with nitrogen, Pd(dppf)Cl₂ (0.13 g, 0.18 mmol, 0.1 eq) was added. The system was heated to 90 °C and stirred for 12 hours under nitrogen protection. After cooling to room temperature, the system was diluted with water and extracted with ethyl acetate. The organic phase was concentrated under reduced pressure and purified by column chromatography to give compound 32 in a total of 0.45 g (yield 66%). 1 H NMR (400MHz, MeOD-d4) δ9.03(s,1H),8.35(d,J=2.4Hz,1H),8.12(d,J=8.0,2.8Hz,1H),7.71(dd,J=8.0,1.6Hz,1H),7.53(dd,J=7.6,1.6 Hz,1H),7.37(t,J=8.0Hz,1H),7.23-7.20(m,1H),3.83(s,3H),3.14-3.01(m,5H),1.85-1.80(m,2H),1.63-1.53(m,2H).LRMS(ESI)calcd for C 19 H 20 FN6O2[M+H] + 383.2, found:383.2.

[0286] Example 33

[0287] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)formamide (compound 33)

[0288] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)formamide

[0289]

[0290] The synthetic route for compound 33 is shown above:

[0291] In step a), compound 33 (0.6 g, 1.57 mmol, 1.0 eq) and reduced iron powder (0.88 g, 15.7 mmol, 10.0 eq) were dissolved in a mixed solvent of EtOH (9 mL) and H2O (1 mL), and then NH4Cl (0.83 g, 15.7 mmol, 10.0 eq) was added. The reaction system was heated to 70 °C and stirred for 3 h. After the reaction was completed, DCM (20 mL) was added to dilute the system, and the mixture was filtered and concentrated under reduced pressure to obtain the crude product MC91 (500 mg).

[0292] Step b: At room temperature, MC91 (25 mg, 0.071 mmol, 1.0 eq) was dissolved in Ac2O (0.1 mL) and HCOOH (0.3 mL) solutions. The system was heated to 60 °C and stirred for 2 hours. After the reaction was completed, the solvent was removed by rotary evaporation under reduced pressure, and compound 33 was purified by prep-HPLC to obtain a white solid (14 mg, 51.7% yield). 1 H NMR(400MHz,MeOD-d4)δ9.18-9.11(m,1H),8.50(s,1H),8.26-8.20(m,1H),7.99-7.91(m,1H),7.26-7.17(m,2 H),7.04-6.90(m,1H),3.85(s,3H),3.09-2.94(m,3H),2.65-2.61(m,2H),2.12-1.91(m,4H).LRMS(ESI)calcd for C 20 H 22 FN6O[M+H] + 381.2, found:381.2.

[0293] Example 34

[0294] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)acetamide (compound 34)

[0295] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)acetamide

[0296]

[0297] The synthetic route for compound 34 is shown above: MC91 (25 mg, 0.071 mmol, 1.0 eq) was dissolved in DCM (2.0 mL) at 0 °C. AcCl (8 mg, 0.102 mmol, 1.3 eq) and TEA (10 mg, 0.102 mmol, 1.3 eq) were added to the system with stirring. The mixture was heated to room temperature and the reaction was continued for 2 hours. After the reaction was completed, the solvent was removed by rotary evaporation under reduced pressure, and then compound 35 was obtained as a white solid (10 mg, 35.6% yield) by prep-HPLC. 1 H NMR(400MHz,MeOD-d4)δ8.91(brs,1H),8.21(brs,1H),7.96(brs,1H),7.24-7.16(m,3H),6.99(brs,1H ),3.80(s,3H),3.06-2.95(m,3H),2.70-2.57(m,2H),2.24(s,3H),2.04-1.90(m,4H).LRMS(ESI)calcd forC 21 H 24 FN6O[M+H] + 395.2, found: 395.3.

[0298] Example 35

[0299] 1,1,1-Trifluoro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide (compound 35)

[0300] 1,1,1-trifluoro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide

[0301]

[0302] The synthetic route is shown above, and the synthetic method is the same as that for compound 33. The data for compound 35 are as follows: 1 H NMR(400MHz,MeOD-d4)δ9.11(s,1H),8.26(brs,1H),8.01(brs,1H),7.39-7.17(m,4H ),3.86(s,3H),3.21-2.99(m,4H),2.66(brs,1H),2.22-1.85(m,4H).LRMS(ESI)calcd forC20 H 21 F4N6O2S[M+H] + 485.1, found: 485.2.

[0303] Example 36

[0304] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide (compound 36)

[0305] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide

[0306]

[0307] The synthetic route for compound 36 is shown above. The crude MC91 product obtained above (30 mg, 0.09 mmol, 1.0 eq) was dissolved in DCM (2.0 mL), and methanesulfonyl chloride (16 mg, 0.14 mmol, 1.5 eq) and TEA (18 mg, 0.18 mmol, 1.5 eq) were added at 0 °C. The reaction was heated to room temperature and stirred for 2 hours. After the reaction was complete, the solvent was removed under reduced pressure, and the product was purified by prep-HPLC to obtain compound 36 as a yellow solid product (20 mg, yield 54%). 1 H NMR(400MHz,MeOD-d4)δ9.12(s,1H),8.22(s,1H),7.97(brs,1H),7.55-7.46(m,1H),7.26(t,J=8.0Hz,1H),7.20-7.17(m ,1H),6.95(brs,1H),3.87(s,3H),3.16(s,3H),3.10-2.99(m,3H),2.69-2.64(m,2H),2.04-1.94(m,4H).LRMS(ESI)calcd forC 20 H 24 FN6O2S[M+H] + 431.2,found:431.2.

[0308] Example 37

[0309] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)cyclopropylsulfonamide (compound 37)

[0310] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)cyclopropanesulfonamide

[0311]

[0312] The synthesis method was the same as in Example 33, and the data for compound 37 were as follows: 1 H NMR(600MHz,MeOD-d4)δ8.98(s,1H),8.22(s,1H),7.97(brs,1H),7.71-7.46(m,1H),7.25(t,J=7.8Hz,1H),7.18(d,J=8.4Hz, 1H),6.94(brs,1H),3.81(s,3H),3.10-2.96(m,3H),2.75-2.65(m,3H),2.04-1.93(m,4H),1.16-1.07(m,4H).LRMS(ESI)calcd for C 22 H 26 FN6O2S[M+H] + 457.2, found: 457.3.

[0313] Example 38

[0314] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)ethylsulfonamide (compound 38)

[0315] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)ethanesulfonamide

[0316]

[0317] The synthetic route is shown above, and the synthetic method is the same as in Example 33. Compound 38 was obtained as a white solid (8 mg, 17% yield), and the structural characterization data are as follows: 1H NMR(600MHz,MeOD-d4)δ9.11(s,1H),8.21(s,1H),7.95(brs,1H),7.56(brs,1H),7.25(t,J=7.8Hz,1H),7.19(d,J=7.8Hz,1H),6.9 2(brs,1H),3.85(s,3H),3.31-3.26(m,2H),3.09-2.99(m,3H),2.65(brs,2H),1.99-1.93(m,4H),1.39(brs,3H).LRMS(ESI)calcd for C 21 H 26 FN6O2S[M+H] + 445.2, found:445.3.

[0318] Example 39

[0319] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)propyl-1-sulfonamide (compound 39)

[0320] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)propane-1-sulfonamide

[0321]

[0322] The synthetic route is shown above, and the synthetic method is the same as in Example 33. The data for compound 39 are as follows: 1 H NMR(600MHz,MeOD-d4)δ9.03(s,1H),8.21(s,1H),7.95(brs,1H),7.56(brs,1H),7.25(t,J=7.8Hz,1H),7.19(d,J=8.4Hz,1H),6.92 (brs,1H),3.83(s,3H),3.27(brs,2H),3.08-2.96(m,3H),2.64(brs,2H),1.98-1.87(m,6H),1.08(t,J=7.2Hz,3H).LRMS(ESI)calcd forC 22 H 28 FN6O2S[M+H] + 459.2, found:459.3.

[0323] Example 40

[0324] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)pyridine-2-sulfonamide (compound 40)

[0325]

[0326] The synthetic route is shown above, and the synthetic method is the same as in Example 33. The data for compound 40 are as follows: 1 H NMR(600MHz,MeOD-d4)δ9.15(s,1H),8.96(s,1H),8.82-8.80(m,1H),8.25-7.88(m,3H),7.66-7. 44(m,2H),7.17-7.01(m,3H),3.87(s,3H),3.32-2.54(m,5H),2.04-1.88(m,4H).LRMS(ESI)calcd for C 24 H 25 FN7O2S[M+H] + 494.2, found: 494.3.

[0327] Example 41

[0328] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-2-methylpropyl-1-sulfonamide (compound 41)

[0329] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-2-methylpropane-1-sulfonamide

[0330]

[0331]

[0332] The synthetic route is shown above, and the synthetic method is the same as in Example 33. The data for compound 41 obtained are as follows: 1H NMR(600MHz,MeOD-d4)δ9.11(s,1H),8.21(s,1H),7.95(brs,1H),7.55(brs,1H),7.26(t,J=7.8Hz,1H),7.19(d,J=8.4Hz,1H),6.92(brs,1H ),3.85(s,3H),3.20(brs,2H),3.08-2.98(m,3H),2.64(brs,2H),2.29(brs,1H),1.98-1.93(m,4H),1.12(d,J=7.2Hz,6H).LRMS(ESI)calcd for C 23 H 30 FN6O2S[M+H] + 473.2, found: 473.3.

[0333] Example 42

[0334] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)butyl-1-sulfonamide (compound 42)

[0335] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)butane-1-sulfonamide

[0336]

[0337] The synthetic route is shown above, and the synthetic method is the same as in Example 33. The data for compound 42 are as follows: 1 H NMR(600MHz,MeOD-d4)δ9.10(s,1H),8.21(s,1H),7.96(brs,1H),7.57(brs,1H),7.26(t,J=7.8Hz,1H),7.19(d,J=7.8Hz,1H),6.92(brs, 1H),3.85(s,3H),3.31-3.26(m,2H),3.09-2.98(m,3H),2.65(brs,2H),1.99-1.81(m,6H),1.49(brs,2H),0.95(brs,3H).LRMS(ESI)calcd for C 23 H 30 FN6O2S[M+H] + 473.2, found: 473.3.

[0338] Example 43

[0339] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)propyl-2-sulfonamide (compound 43)

[0340] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)propane-2-sulfonamide

[0341]

[0342] The synthetic route is shown above, and the synthetic method is the same as in Example 33. The data for compound 43 are as follows: 1 H NMR(600MHz,MeOD-d4)δ9.07-8.96(m,1H),8.21(s,1H),7.96-7.93(m,1H),7.60(brs,1H),7.24(t,J=7.8Hz,1H),7.21-7.18(m,1H),6. 90(brs,1H),3.81(s,3H),3.50(brs,1H),3.09-2.94(m,3H),2.69-2.63(m,2H),1.98-1.89(m,4H),1.43-1.41(m,6H).LRMS(ESI)calcd forC 22 H 28 FN6O2S[M+H] + 459.2, found:459.3.

[0343] Example 44

[0344] 3-Chloro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)propyl-1-sulfonamide (Compound 44)

[0345] 3-chloro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)propane-1-sulfonamide

[0346]

[0347] The synthetic route is shown above, and the synthetic method is the same as in Example 33. The data for compound 44 are as follows: 1 H NMR(600MHz,MeOD-d4)δ9.02(s,1H),8.22(s,1H),7.95(brs,1H),7.56(brs,1H),7.26(t,J=7.8Hz,1H),7.19(d,J=8.4Hz,1H),6.94( LRMS(ESI)calcd forC 22 H 27 ClFN6O2S[M+H] + 493.2, found: 493.3.

[0348] Example 45

[0349] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-N-methylmethanesulfonamide (compound 45)

[0350] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-N-methylmethanesulfonamide

[0351]

[0352] The synthetic method is as described above. Compound 36 (20 mg, 0.05 mmol, 1.0 eq) was dissolved in THF (2.0 mL), and potassium carbonate (14 mg, 0.10 mmol, 2.0 eq) and methyl iodide (14 mg, 0.10 mmol, 2.0 eq) were added at room temperature. The reaction was stirred at room temperature for 2 hours. After the reaction was completed, the solvent was removed under reduced pressure, and the product was separated and purified by prep-HPLC to obtain compound 45 as a white solid product (8 mg, yield 40%). The data for compound 45 are as follows: 1H NMR(400MHz,MeOD-d4)δ9.11(s,1H),8.29(s,1H),8.05(brs,1H),7.50-7.45(m,1H),7.29-7.26(m,2H ),7.19-7.15(m,1H),3.86(s,3H),3.33(s,3H),3.09-2.96(m,7H),2.04-1.66(m,5H).LRMS(ESI)calcd for C 21 H 26 FN6O2S[M+H] + 445.2, found:445.3.

[0353] Example 46

[0354] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)thiophene-2-sulfonamide (compound 46)

[0355] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)thiophene-2-sulfonamide

[0356]

[0357] The synthetic route is shown above, and the synthetic method is the same as in Example 33. The data for compound 46 are as follows: 1 H NMR(600MHz,MeOD-d4)δ9.02(s,1H),8.13(brs,1H),7.89-7.79(m,2H),7.63(brs,2H), 7.26-6.92(m,4H),3.82(s,3H),3.03-2.53(m,5H),1.91-1.88(m,4H).LRMS(ESI)calcd forC 23 H 24 FN6O2S2[M+H] + 499.1, found: 499.3.

[0358] Example 47

[0359] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methylamino-1-sulfonamide (compound 47)

[0360] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methylamino-1-sulfonamide

[0361]

[0362] The synthetic route is shown above, and the synthetic method is the same as in Example 33. The data for compound 47 are as follows: 1 H NMR(600MHz,MeOD-d4)δ9.09(s,1H),8.20(s,1H),7.94(brs,1H),7.54(d,J=7.2Hz,1H),7.25(t,J=7.8Hz,1H),7.19(d ,J=8.4Hz,1H),6.86(brs,1H),3.85(s,3H),3.09-2.96(m,3H),2.80-2.65(m,5H),2.04-1.93(m,4H).LRMS(ESI)calcd for C 20 H 25 FN7O2S[M+H] + 446.2, found:446.3.

[0363] Example 48

[0364] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-4-methylbenzenesulfonamide (Compound 48)

[0365] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-4-methylbenzenesulfonamide

[0366]

[0367] The synthetic route is shown above, and the synthetic method is the same as in Example 33. The data for compound 48 are as follows: 1H NMR(400MHz,MeOD-d4)δ9.16(s,1H),8.10(brs,1H),7.85(brs,1H),7.72(d,J=7.6Hz,2H),7.56(brs,1H),7.36(d,J=8.0H z,2H),7.16-7.13(m,2H),6.88(brs,1H),3.86(s,3H),3.00-2.49(m,5H),2.41(s,3H),1.93-1.89(m,4H).LRMS(ESI)calcd for C 26 H 28 FN6O2S[M+H] + 507.2, found:507.2.

[0368] Example 49

[0369] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-2-methoxyethyl-1-sulfonamide (compound 49)

[0370] N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-2-methoxyethane-1-sulfonamide

[0371]

[0372] The synthetic route is shown above, and the synthetic method is the same as in Example 33. The data for compound 49 are as follows: 1 H NMR(600MHz,MeOD-d4)δ9.15(s,1H),8.20(s,1H),7.95(brs,1H),7.61(brs,1H),7.25(t,J=7.8Hz,1H),7.18(d,J=8.4Hz,1H),6.90(brs ,1H),3.86(s,3H),3.84(brs,2H),3.55(brs,2H),3.27(s,3H),3.08-2.98(m,3H),2.63(brs,2H),1.99-1.93(m,4H).LRMS(ESI)calcdfor C 22 H 28 FN6O3S[M+H] + 475.2, found:475.2.

[0373] Example of effect 1

[0374] The method for testing the QPCTL enzyme inhibitory activity of the compound is as follows: The reaction system consisted of QPCTL protease, the fluorescent substrate glutamine 7-amino-4-methylcoumarin (Gln-AMC), the compound, and pyroglutamylaminopeptidase I (PGPEP I). The reaction was carried out in a black 384-well plate. First, 12.5 μL of QPCTL protease (7 ng / μL) and 2.5 μL of the compound at different concentrations were mixed and reacted in a shaker at 37°C and 100 rpm for 10 minutes. Then, 10 μL of the fluorescent substrate Gln-AMC (final concentration 200 μM) was added, and the reaction was carried out in a shaker at 37°C and 100 rpm for 20 minutes. Finally, 25 μL of PGPEP I (final concentration 2.87 ng / μL) was added to the system, and the reaction was carried out in a shaker at 37°C and 100 rpm for 30 minutes. The fluorescence intensity of the microplates was read using a TECANinfinite 200Pro microplate reader at excitation / emission wavelengths of 380 / 460 nm. The obtained fluorescence signal values ​​were analyzed using GraphPad Prism 8.0 to obtain the inhibition rate and IC50 of the compounds. 50 The activity data are shown in Table 1 below:

[0375] Table 1. Activity data of representative compounds

[0376] Compound numbering <![CDATA[Enzyme activity IC 50 (nM)]]> Compound numbering <![CDATA[Enzyme activity IC 50 (nM)]]> PQ912 54.6 Compound 25 >1000 SEN177 131.2 Compound 26 >1000 Compound 1 >1000 Compound 27 >1000 Compound 2 >1000 Compound 28 458.2 Compound 3 >1000 Compound 29 292.1 Compound 4 >1000 Compound 30 1.3 Compound 5 >1000 Compound 31 28.4 Compound 6 94.3 Compound 32 5.4 Compound 7 >1000 Compound 33 34.3 Compound 8 >1000 Compound 34 478.2 Compound 9 50 Compound 35 18.8 Compound 10 >1000 Compound 36 1.4 Compound 11 >1000 Compound 37 3.6 Compound 12 6.2 Compound 38 1.5 Compound 13 >1000 Compound 39 3.9 Compound 14 >1000 Compound 40 1.9 Compound 15 19.8 Compound 41 2.5 Compound 16 7.6 Compound 42 2.6 Compound 17 >1000 Compound 43 0.6 Compound 18 719.9 Compound 44 33.3 Compound 19 365.9 Compound 45 59 Compound 20 >1000 Compound 46 1.7 Compound 21 33.9 Compound 47 4.0 Compound 22 19.6 Compound 48 0.4 Compound 23 11.6 Compound 49 3.5 Compound 24 505.6

[0377] Based on the above experimental results, it can be seen that the compound of the present invention has excellent QPCTL enzyme inhibitory activity and can be used as a glutamine cyclase inhibitor. Its anti-tumor effect will be further studied.

[0378] Example 2

[0379] The method for testing the cellular activity of the compound is as follows:

[0380] The inhibitory activity of the compound against N-terminal pyroglutamate modification (pGlu-CD47) of CD47 cells was detected as follows: 293T cells were seeded at a density of 10,000 / well in 48-well plates. After treatment with different concentrations of the compound for 48 hours, the in vitro activity of the compound was tested using flow cytometry (FACS). The level of N-terminal pyroglutamate modification (pGlu-CD47) of CD47 on the cell surface was detected using the CD47 flow cytometry antibody CC2C6. After treatment with the compound, the cells were washed with PBS and resuspended in 100 μL of FACS buffer. Then, 1 μL of cell flow cytometry antibody was added, and the cells were incubated on ice in the dark for 30 minutes. After washing with FACS buffer, the cells were resuspended in 200 μL of FACS buffer. The cells were analyzed by flow cytometry, and the mean fluorescence intensity (MFI) was analyzed using GraphPad Prism 8.0 to calculate the pGlu-CD47 inhibition rate of the compound. The activity data of representative compounds are shown in Table 2 below.

[0381] Table 2. Cell activity data representing compounds

[0382]

[0383] Experimental results show that the compound of the present invention has a good inhibitory effect on the N-terminal pyroglutamate modification of CD47 on the surface of 293T cells and has extremely high cell activity.

[0384] PQ912 (CAS: 1276021-65-8) and SEN177 (CAS: 2117405-13-5) are both known glutamine acyl cyclase inhibitors. As positive control drugs, they demonstrate that the compounds of the present invention have better enzyme inhibitory activity and antitumor activity, and demonstrate that the compounds of the present invention have better drug development potential.

[0385] Those skilled in the art should understand that variations can be implemented by combining existing technology with the above embodiments, which will not be elaborated here. Such variations do not affect the essence of the present invention, and will not be elaborated here either.

[0386] The preferred embodiments of the present invention have been described above. It should be understood that the present invention is not limited to the specific embodiments described above, and the devices and structures not described in detail should be understood as being implemented in a conventional manner in the art. Any person skilled in the art can make many possible variations and modifications to the technical solutions of the present invention using the methods and techniques disclosed above, or modify them into equivalent embodiments with equivalent changes, without departing from the scope of the present invention. This does not affect the essential content of the present invention. Therefore, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the present invention's technical solutions still fall within the protection scope of the present invention.

Claims

1. A nitrogen-containing heterocyclic compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof: in: ---- indicates a single key or that the key does not exist; m is any integer between 0 and 3, and n is any integer between 0 and 3; R1 is a 5-10 heteroaryl group optionally substituted with one or more R7s; R2 is cyano, nitro, -C(=O)N(R8)2, -NR8C(=O)R9, -S(=O)2R9 or -NR8S(=O)2R9; R3 is a hydrogen atom or a halogen atom; R4is optionally substituted with one or more C 1-20 alkyl substituted 5-10 membered heteroaryl; R5 represents a hydrogen atom, a halogen atom, a hydroxyl group, or a carbon atom. 1-20 Alkyl or C 1-20 Alkoxy; optionally, when R5 is C 1-20 In the case of alkyl groups, any carbon atom of R5 is connected to any ring atom of R4 to form a 5-10 membered ring structure; R6 represents a hydrogen atom or -NR8C(=O)R9; Each R7 is independently a halogen atom, arbitrarily C-treated. 3-10 Cycloalkyl-substituted C 1-20 Alkyl, optionally C 3-10 Cycloalkyl-substituted C 1-20 Alkoxy or optionally C 1-20 Alkyl-substituted C 3-10 Cycloalkyloxy; Each R8 atom is independently either a hydrogen atom or a carbon atom. 1-20 alkyl; Each R9 atom is independently composed of hydrogen and carbon atoms. 1-20 Alkyl, C 1-20 Halogenated alkyl groups, -NHC 1-20 Alkyl, -C 1-20 Alkylene-C 1-20 Alkoxy, C 3-10 Cycloalkyl, 5-10 membered heterocycloalkyl, 5-10 membered heteroaryl or C 6-10 Aryl; the C 3-10 Cycloalkyl, 5-10 membered heterocycloalkyl, 5-10 membered heteroaryl or C 6-10 Each aryl group is independently and optionally bonded by one or more halogen atoms, C 1-20 Alkyl or -C(=O)OC 1-20 Alkyl substitution.

2. The compound according to claim 1, or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug, characterized in that, m is 0 or 1, n is 0 or 1; R1 is a 5-10 nitrogen-containing heteroaryl group optionally substituted with one or more R7s; R2 is cyano, nitro, -C(=O)N(R8)2, -NR8C(=O)R9, -S(=O)2R9 or -NR8S(=O)2R9; R3 is a hydrogen atom or a halogen atom; R4 is arbitrarily assigned to one or more Cs 1-10 Alkyl-substituted 5-10 nitrogen-containing heteroaryl groups; R5 represents a hydrogen atom, a halogen atom, a hydroxyl group, or a carbon atom. 1-10 Alkyl or C 1-10 Alkoxy; optionally, when R5 is C 1-10 In the case of alkyl groups, any carbon atom of R5 is connected to any ring atom of R4 to form a 5-6 membered ring structure; R6 represents a hydrogen atom or -NR8C(=O)R9; Each R7 is independently a halogen atom, arbitrarily C-treated. 3-6 Cycloalkyl-substituted C 1-10 Alkyl, optionally C 3-6 Cycloalkyl-substituted C 1-10 Alkoxy or optionally C 1-10 Alkyl-substituted C 3-6 Cycloalkyloxy; Each R8 atom is independently either a hydrogen atom or a carbon atom. 1-10 alkyl; Each R9 atom is independently composed of hydrogen and carbon atoms. 1-10 Alkyl, C 1-10 Halogenated alkyl groups, -NHC 1-10 Alkyl, -C 1-10 Alkylene-C 1-10 Alkoxy, C 3-6 Cycloalkyl, 5-6 membered heterocycloalkyl, 5-10 membered heteroaryl or C 6-10 Aryl; the C 3-6 Cycloalkyl, 5-6 membered heterocycloalkyl, 5-10 membered heteroaryl or C 6-10 Each aryl group is independently and optionally bonded by one or more halogen atoms, C 1-10 Alkyl or -C(=O)OC 1-10 Alkyl substitution.

3. The compound according to claim 2, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof, characterized in that, m is 0 or 1, n is 0 or 1, and m and n are not both 0 at the same time; R1 is a pyridinyl group optionally substituted with one or more R7 groups; R2 is cyano, nitro, -C(=O)N(R8)2, -NR8C(=O)R9, -S(=O)2R9 or -NR8S(=O)2R9; R3 is a hydrogen atom or a halogen atom; R4 is arbitrarily assigned to one or more Cs 1-10 Alkyl-substituted groups include: in, Indicates the combination of keys; R5 is a hydrogen atom, a halogen atom, a hydroxyl group, or a carbon atom. 1-10 Alkyl; optionally, when R5 is C 1-10 In the case of alkyl groups, any carbon atom of R5 is connected to any ring atom of R4 to form a 5-6 membered ring structure; R6 represents a hydrogen atom or -NR8C(=O)R9; Each R7 is an independent halogen atom; Each R8 atom is independently either a hydrogen atom or a carbon atom. 1-10 alkyl; Each R9 atom is independently composed of hydrogen and carbon atoms. 1-10 Alkyl, C 1-10 Halogenated alkyl groups, -NHC 1-10 Alkyl, -C 1-10 Alkylene-C 1-10 Alkoxy, C 3-6 Cycloalkyl, 5-6 membered heterocycloalkyl, 5-10 membered heteroaryl or C 6-10 Aryl; the C 3-6 Cycloalkyl, 5-6 membered heterocycloalkyl, 5-10 membered heteroaryl or C 6-10 Each aryl group is independently and optionally bonded by one or more halogen atoms, C 1-10 Alkyl or -C(=O)OC 1-10 Alkyl substitution.

4. The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof, characterized in that, The compound is shown in formula (II): Wherein, m, n, R1, R2, R3, R4, R5 and R6 have the definitions as described in any one of claims 1-3.

5. The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof, characterized in that, The compound is shown in formula (III): Wherein, m, n, R1, R2, R3 and R6 have the definitions as described in any one of claims 1-3; p is any integer from 0 to 3, preferably 1 or 2.

6. The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof, characterized in that, The compound is shown in formula (IV): Wherein, m, n, R1, R2, R3, R4, R5 and R6 have the definitions as described in any one of claims 1-3.

7. The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof, characterized in that, The compound is shown in formula (V): Wherein, m, n, R2, R4, R5, R8 and R9 have the definitions as described in any one of claims 1-3; Preferably, R4 is arbitrarily determined by one or two Cs. 1-10 Alkyl-substituted groups include: in, Indicates the combination of keys; Preferably, R5 is a hydrogen atom or a halogen atom.

8. The following nitrogen-containing heterocyclic compounds or their pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, isotope-labeled substances, or prodrugs:

9. A pharmaceutical composition comprising, as an active ingredient, a nitrogen-containing heterocyclic compound according to any one of claims 1-8 or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof; preferably, the pharmaceutical composition contains at least one pharmaceutically acceptable carrier.

10. A combination of drugs comprising a nitrogen-containing heterocyclic compound according to any one of claims 1-8 or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug, or a pharmaceutical composition according to claim 9, and an antibody; preferably, the antibody is a PD-1 / PD-L1 antibody or a CD47 antibody.

11. The use of the nitrogen-containing heterocyclic compound or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label or prodrug according to any one of claims 1-8, or the pharmaceutical composition according to claim 9, or the combination of drugs according to claim 10, in the inhibition of glutamine cyclase.

12. The use of the nitrogen-containing heterocyclic compound according to any one of claims 1-8, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof, or the pharmaceutical composition according to claim 9, or the combination of drugs according to claim 10, in the preparation of a medicament for the prevention and / or treatment of tumors, immune diseases, neurological diseases, or aging; Preferably, the tumor is selected from at least one of colorectal cancer, lung cancer, gastric cancer, melanoma, myeloma, breast cancer, adenocarcinoma, bladder cancer, and hematologic malignancy; Preferably, the immune disease is selected from at least one of eczema, alopecia areata, psoriasis, vitiligo, rheumatoid arthritis, lupus erythematosus syndrome, acne, and hidradenitis suppurativa; Preferably, the neurological disease is selected from at least one of Alzheimer's disease, Huntington's disease, Down syndrome, depression, anxiety disorder, psychosis, and multiple sclerosis.

Images