WO2025247243A1 - Nitrogen-containing heterocyclic compound and use thereof - Google Patents

Abstract

Disclosed in the present invention are a nitrogen-containing heterocyclic compound and use thereof. The compound has a structure represented by formula (I). The compound of the present invention has an excellent inhibitory effect on glutaminyl cyclase, can be used in treating a tumor, an immunological disease, a neurological disease, obesity, or aging, and has good application prospects.

Description

A nitrogen-containing heterocyclic compound and its application Technical Field

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

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.

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.

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

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, W.H.J. et al. 1987, J Biol Chem 262, 8532-8536; Fischer, W.H. 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, W.H.J. et al. 1987, J Biol Chem 262, 8532-8536; Fischer, W.H. 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, T.M. et al. 1995, J Neuroendocrinol 7, 445-453). It has been confirmed that the enzyme activity of recombinant human QPCT and QPCT derived from brain extracts both catalyze the cyclization of N-terminal glutamine and glutamate. Most surprisingly, it was found that approximately pH 6.0 favored the cyclase-catalyzed Glu-conversion, while the conversion of Gln- to pGlu-derived compounds occurred at an optimal pH of approximately 8.0. Since inhibiting the activity of recombinant human QPCT and QPCT-derived from porcine pituitary extracts 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 Res Ther 2018, 10, 107).

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).

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, and phosphatases SHP-1/SHP-2 are recruited and activated, 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.

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.

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, 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.

Besides CD47, QPCT and QPCTL can also pyroglutamate other proteins. For example, β-amyloid protein, which is known to be involved in Alzheimer's disease, is pyroglutamate-modified by QPCT (Pharmacological Research, 2019 147, 104342), while C-C motif chemokine ligand 2 (CCL2) protein is pyroglutamate-modified by QPCTL (EMBO Mol Med, 2011, 3, 510-512). In addition, the N-terminus of CX3CL1 can also be pyroglutamate-modified by QPCT and QPCTL (Bioscience Reports, 2017 37, BSR20170712). Recent studies have found that QPCTLs 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, a certain degree of functional overlap between QPCTs and QPCTLs in the pyroglutamate ...

However, there are still no targeted drugs available for QPCT or QPCTL inhibitors.

Therefore, developing a new QPCT or QPCTL inhibitor to provide new treatments for tumors, immune diseases, neurological diseases, obesity, or aging is of great practical significance.

Summary of the Invention

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

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

This invention provides 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 from 0 to 3, and n is any integer from 0 to 3;

X is a carbon atom or a nitrogen atom;

_R1 is a 5-10 heteroaryl group optionally substituted by one or more _R7s ;

_R2 can be cyano, nitro, -C(=O)N( _R8 ) ₂ , _-NR8C (=O) _R9 , -S(=O) _{2R9 or -NR8S} (=O) _{2R9 ;}

_R3 is a hydrogen atom, a halogen atom, a trifluoromethyl atom, a -NR ₈ C(=O)R ₉ , or a C _1-20 alkoxy atom;

_R4 is a 5-10 heteroaryl group optionally substituted with one or more _C1-20 alkyl groups;

_R5 is a hydrogen atom, a halogen atom, a hydroxyl group, a _C1-20 alkyl group, or a _C1-20 alkoxy group; optionally, when _R5 is a _C1-20 alkyl group, 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 R ₇ is independently a halogen atom, a C _1-20 alkyl atom optionally substituted with a C _3-10 cycloalkyl atom, a C _1-20 alkoxy atom optionally substituted with a C _3-10 cycloalkyl atom, or a C _3-10 cycloalkyloxy atom optionally substituted with a C _1-20 alkyl atom;

Each R ₈ is independently a hydrogen atom or a _C1-20 alkyl group;

Each R ₉ is independently a hydrogen atom, a _C1-20 alkyl, a _C1-20 haloalkyl, a _-NHC1-20 alkyl, a _-C1-20 alkylene- _C1-20 alkoxy, a _C3-10 cycloalkyl, a 5-10 heterocyclic alkyl, a 5-10 heteroaryl, or a _C6-10 aryl; the _C3-10 cycloalkyl, 5-10 heterocyclic alkyl, 5-10 heteroaryl, or _C6-10 aryl is independently optionally substituted by one or more halogen atoms, a _C1-20 alkyl, or a -C(=O) _OC1-20 alkyl; or, R ₈ and R ₉ are linked to form a 5-10 cyclic structure.

In some preferred embodiments, in formula (I):

m is 0 or 1, n is 0 or 1;

X is a carbon atom or a nitrogen atom;

_R1 is a 5-10 nitrogen-containing heteroaryl group optionally substituted by one or more _R7 ;

_R2 can be cyano, nitro, -C(=O)N( _R8 ) ₂ , _-NR8C (=O) _R9 , -S(=O) _{2R9 or -NR8S} (=O) _{2R9 ;}

_R3 is a hydrogen atom, a halogen atom, a trifluoromethyl atom, a -NR ₈ C(=O)R ₉ or a C _1-10 alkoxy atom;

_R4 is a 5-10 nitrogen-containing heteroaryl group optionally substituted with one or more _C1-10 alkyl groups;

_R5 is a hydrogen atom, a halogen atom, a hydroxyl group, a _C1-10 alkyl group, or a _C1-10 alkoxy group; optionally, when _R5 is a _C1-10 alkyl group, 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 R ₇ is independently a halogen atom, a _C1-10 alkyl group, a _C1-10 alkyl group optionally substituted with a _C3-6 cycloalkyl group, a _C1-10 alkoxy group optionally substituted with a _C3-6 cycloalkyl group, or a _C3-6 cycloalkyloxy group optionally substituted with a _C1-10 alkyl group.

Each R ₈ is independently a hydrogen atom or a _C1-10 alkyl group;

Each R ₉ is independently a hydrogen atom, a C _1-10 alkyl, a C _1-10 haloalkyl, a -NHC _1-10 alkyl, a -C _1-10 alkylene-C _1-10 alkoxy, a C _3-6 cycloalkyl, a 5-6 membered heterocyclic alkyl, a 5-10 membered heteroaryl, or a C _6-10 aryl; the C _3-6 cycloalkyl, 5-6 membered heterocyclic alkyl, 5-10 membered heteroaryl, or C _6-10 aryl are each optionally substituted by one or more halogen atoms, C _1-10 alkyl, or -C(=O)OC _1-10 alkyl; or, R ₈ and R ₉ are linked to form a 5-6 membered ring structure.

In some preferred embodiments, in formula (I):

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

X is a carbon atom or a nitrogen atom;

_R1 is a pyridinyl group optionally substituted with one or more _R7 groups;

_R2 can be cyano, nitro, -C(=O)N( _R8 ) ₂ , _-NR8C (=O) _R9 , -S(=O) _{2R9 or -NR8S} (=O) _{2R9 ;}

_R3 is a hydrogen atom, a halogen atom, a trifluoromethyl atom, a -NR ₈ C(=O)R ₉ , or a C _1-10 alkoxy atom;

_R4 is any of the following groups optionally substituted with one or more _C1-10 alkyl groups: in, Indicates the combination of keys;

_R5 is a hydrogen atom, a halogen atom, a hydroxyl group, or a _C1-10 alkyl group; optionally, when _R5 is a _C1-10 alkyl group, 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 R ₇ is independently substituted with a methyl, methoxy, or cyclopropyl methoxy or halogen atom;

Each R ₈ is independently a hydrogen atom or a _C1-10 alkyl group;

Each _R9 is independently a hydrogen atom, a _C1-10 alkyl, a _C1-10 haloalkyl, a _-NHC1-10 alkyl, a _-C1-10 alkylene- _C1-10 alkoxy, a _C3-6 cycloalkyl, a 5-6 heterocyclic alkyl, a 5-10 heteroaryl, or a _C6-10 aryl; the _C3-6 cycloalkyl, 5-6 heterocyclic alkyl, 5-10 heteroaryl, or _C6-10 aryl are each optionally substituted by one or more halogen atoms, _C1-10 alkyl, or -C(=O) _OC1-10 alkyl; or, _R8 and _R9 are linked to form a five-membered ring structure.

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

Wherein, m, n, X, _R1 , _R2 , _R3 , _R4 , _R5 and _R6 have the definitions described in equation (I).

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

Wherein, m, n, X, _R1 , _R2 , _R3 and _R6 have the definitions described in equation (I);

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

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

Wherein, m, n, X, _R1 , _R2 , _R3 , _R4 , _R5 and _R6 have the definitions described in equation (I).

In other embodiments, the compound is as shown in formula (V):

In some preferred embodiments, the compound is as shown in formula (VI) or (VII):

Wherein, m, n, X, _R1 , _R4 , _R5 , _R8 and _R9 have the definitions described in equation (I);

Preferably, _R4 is a group optionally substituted with one or two _C1-10 alkyl groups, such as the following groups: in, Indicates the combination of keys;

Preferably, _R5 is a hydrogen atom or a halogen atom. In some preferred embodiments, the compound is as shown in formula (VIII):

Wherein, m, n, X, _R1 , _R2 , _R3 , _R5 and _R6 have the definitions described in equation (I);

Z represents a hydrogen atom or a nitrogen atom;

_R10 is a benzyl group substituted with a hydrogen atom, a _C1-10 alkyl group, or a _C1-10 alkoxy group; optionally, _R5 and _R10 are linked to form a 5-10 membered ring structure.

R ₁₁ is a hydrogen atom, a _C1-10 alkyl group, or an amino group.

Preferably, _R10 is a _C1-10 alkyl group. More preferably, _R10 is a _C1-2 alkyl group. The _C1-2 alkyl group includes methyl, ethyl, and _-CD3 .

Preferably, _R11 is a hydrogen atom, a _C1-5 alkyl group, or an amino group.

In some preferred embodiments, _R4 is Z, _R10 , and _R11 have the definitions described in equation (VIII).

In some preferred embodiments, _R1 is one of the following groups:

in, Indicates the combination of keys;

Each R ₁₂ is an independent halogen atom;

R ₁₃ is a methoxy or halogen atom substituted with methyl, methoxy, or cyclopropyl groups;

Each R ₁₄ is independently a _C1-10 alkyl group.

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

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.

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.

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.

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, obesity or aging.

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.

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.

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.

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.

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.

EC includes QPCT and QPCTL as glutamate cyclase (EC) activities, which are further defined as EC activities.

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).

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.

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

Efficacy of QPCTL inhibition: In view of its relationship with QPCTL inhibition, in a preferred embodiment, the _IC50 of QPCTL inhibition 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.

The present invention has the following advantages or beneficial effects:

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

Detailed Implementation

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

Example 1

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

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

The synthetic route for compound 1 is shown in the figure above.

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). Pd(dppf) _Cl2 (18 mg, 0.024 mmol, 0.1 eq) was added under argon protection. 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, CDCl ₃ ) δ 8.54 (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).

Step b: Intermediate MC3 (1.47 g, 1.0 eq) was dissolved in MeOH (20 mL). Pd/C (10%, 200 mg) was added under argon protection. The system was purged three times with hydrogen at room temperature, and the reaction was carried out overnight at 45 °C. After the reaction was complete, the mixture was filtered, and the organic phase was concentrated to obtain compound MC4 (1.2 g). ^¹H NMR (400 MHz, _CDCl₃ ) δ 8.62 (s, 1H), 7.58 (s, 1H), 4.16–4.11 (m, 2H), 3.03–2.95 (m, 1H), 2.79 (t, J = 13.2 Hz, 2H), 1.95–1.91 (m, 2H), 1.63–1.51 (m, 2H), 1.41 (s, 9H).

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.

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.

The data for compound 1 were as follows: ^¹H NMR (400MHz, MeOD- _d⁴ ) δ 9.03 (s, ¹H), 8.26 (d, J = 2.5 Hz, ¹H), 8.07–7.92 (m, ¹H), 7.83–7.63 (m, 2H), 7.54 (dd, J = 7.7, 1.6 Hz, ¹H), 7.31 (t, J = 7.7 Hz, 1H), 7.20 (dd, J = 8.5, 2.6 Hz, 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.

Example 2

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

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

The synthetic route for compound 2 is shown in the figure above.

In step a), compound MC7 (1.05 g, 5.00 mmol, 1.0 eq), trimethylsilyl azido (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 was heated to 100 °C and stirred for 8 h. After the reaction was completed, EA (50 mL) was added for dilution, followed by washing with water (50 mL) and brine (50 mL), drying with anhydrous sodium sulfate, and then concentrating under reduced pressure. The concentrate was purified by silica gel column chromatography to obtain intermediate MC8 (600 mg, 47.6% yield). ^¹H NMR (400 MHz, _CDCl₃ ) δ 7.47 (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).

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).

Steps c and d: Using intermediate MC9, the reaction proceeds as described in step c of Example 1 to generate intermediate MC10. Then, as described in step d of Example 1, the data for compound 2 are as follows: ^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,1 H),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.

Example 3

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

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

The synthetic route of compound 3 is shown in the figure above.

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). ¹ H NMR (400MHz, CDCl ₃ )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).

Steps b and c: Using intermediate MC13, react as in step c of Example 1 to generate intermediate MC14, then as in step d of Example 1, to obtain the data for compound 3 as follows: ^¹H NMR (400MHz, MeOD-d ₄₎ )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.

Example 4

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

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

The synthesis method was the same as in Example 3, as shown in the route above. The data for compound 4 were as follows: ^¹H NMR (400MHz, MeOD- _d⁴ ) δ 8.95 (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, 1H), 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.

Example 5

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

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

The synthesis method was the same as in Example 3, as shown in the route above. The data for compound 5 were as follows: ^¹H NMR (400MHz, DMSO) δ 8.28 (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.

Example 6

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)

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

The synthesis method was the same as in Example 1, and the synthetic route is shown in the figure above. The data for compound 6 were as follows: ^¹H 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,1H),7.22(d d,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.

Example 7

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

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data of compound 7 are as follows: ^¹H NMR (400MHz, MeOD- _d⁴ ) δ 9.47 (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.

Example 8

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

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data for compound 8 are as follows: ^¹H 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.

Example 9

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

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data for compound 9 are as follows: ^¹H 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 FN ₆ [M+H] ⁺ 377.2, found: 377.3.

Example 10

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

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

The synthetic route of compound 10 is shown in the figure above:

In 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). ^¹H NMR (600 MHz, _CDCl₃ ) δ 7.39–7.32 (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).

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 (400 MHz, _CDCl3 ) δ 8.96 (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).

In step c, intermediates 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 solution was concentrated under reduced pressure and purified by silica gel column chromatography to obtain intermediate MC35 (20 mg, 74.9% yield). ^¹H NMR (600 MHz, _CDCl₃ ) δ 7.44 (s, 1H), 4.00 (s, 3H), 2.84–2.72 (m, 3H), 1.88 (d, J = 13.8 Hz, 2H), 1.72 (brs, 2H), 1.63–1.54 ( _m , 2H), 1.47 (s, 9H) _. LRMS (ESI) calcd _{for C₁₃H₂₃N₄O₂ [} M+H] ^⁺ 267.2, found: 267.3.

After removing the Boc protecting group using compound MC35 in steps d and e, the intermediate MC36 was synthesized using the same method as step c in Example 1. The data for compound 10 obtained using step d in Example 1 are as follows: ^1H 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 FN ₆ [M+H] ⁺ 363.2,found:363.3.

Example 11

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

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

The synthetic route of compound 11 is shown in the figure above:

Step a: Dissolve compound MC37 (814 mg, 11.0 mmol, 1.0 eq) in acetonitrile (5 mL), then add DMFDMA (1.35 g, 11.0 mmol, 1.0 eq) to the system, heat to 50 °C and stir for 1 hour. Then add compound MC38 (1.0 g, 5.5 mmol, 0.5 eq) and HOAc (1 mL) to the system, heat to 100 °C and stir 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). ^1H NMR (600 MHz, _CDCl3 ) δ 8.00 (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).

Steps b and c: Using intermediate MC39, the reaction proceeds as described in step c of Example 1 to generate intermediate MC40. Then, as described in step d of Example 1, compound 11 is synthesized. The data are as follows: ^1H 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 FN ₆ [M+H] ⁺ 363.2,found:363.3.

Example 12

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

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data for compound 12 are as follows: ^¹H NMR (400MHz, MeOD- _d⁴ ) δ 8.87 (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 = 2.4Hz, 1H). ＝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 FN ₅ [M+H] ⁺ 376.2,found:376.3.

Example 13

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

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data for compound 13 are as follows: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 8.57-8.53 (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 _{C₂₃H₂₂} FN ₄ [M+H] ⁺ 373.2,found:373.3.

Example 14

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

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data for compound 14 are as follows: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 8.64-8.62 (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 C₂₃H₂₂FN₄ [ M} +H] ^⁺ 373.2, found: 373.3.

Example 15

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

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data for compound 15 are as follows: ^¹H 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),3.11(brs,2H),2.28(brs,2H),1.86-1.84(m,2H),1.48(s,3H).LRMS(ESI)[M+H] ⁺ ,found:377.2.

Example 16

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

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data of compound 16 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 8.75 (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.

Example 17

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)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data for compound 17 are as follows: ^¹H 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.

Example 18

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

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data of compound 18 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 9.10 (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.

Example 19

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

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

The synthetic route is shown in the figure above. 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,1H),7.69(dd,J＝8.0,1.6Hz,1H),7.35(dd,J＝7.6,1.6Hz,1H),7.30(t,J＝7.6Hz,1H),7.1 7-7.13(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 FN ₆ [M+H] ⁺ 349.2, found:349.3.

Example 20

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

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data for compound 20 are as follows: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 9.05 (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, 1.6Hz, 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 for C₁₈H₁₆FN₆ [} M+ ^H ] _⁺ 335.1, found: 335.3.

Example 21

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)

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

The synthetic route of compound 21 is shown in the figure above:

In 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), and the reaction system was heated to reflux and stirred overnight. After the reaction was completed, the mixture was concentrated under reduced pressure and purified by silica gel column chromatography to obtain intermediate MC61 (290 mg, 51.1% yield). ^¹H NMR (400 MHz, _CDCl₃ ) δ 8.31 (s, 1H), 4.01 (brs, 2H), 3.34 (brs, 2H), 2.90 (t, J = 11.6 Hz, 2H), 2.54 (t, J = 12.4 Hz, 2H), 1.88 (brs, 4H), 1.52–1.44 (m, 11H). _{LRMS (} ESI) calcd for _{C₁₄H₂₅N₂O₂S} [M+H] ^⁺ _285.2 , found: 285.3.

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), and then AgOBz (460 mg, 2.0 mmol, 2.0 eq) and HOAc (0.17 mL) were added. The reaction system was heated to reflux and stirred overnight. After the reaction was completed, 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 (400 MHz, _CDCl₃ ) δ 8.08 (s, 1H), 4.01 (t, J = 6.0 Hz, 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 C₁₅H₂₅N₄O₂ [} M+H] ^⁺ 293.2, found: 293.3.

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: ^¹H 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),7.34(t,J＝7.6 Hz,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 FN ₆ [M+H] ⁺ 389.2,found:389.3.

Example 22

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)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 21. The data for compound 22 are as follows: ^¹H NMR (400MHz, MeOD- _d⁴ ) δ 9.23 (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 FN ₆ [M+H] ⁺ 375.2,found:375.2.

Example 23

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

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 16. The data for compound 23 are as follows: ^¹H NMR (600MHz, MeOD- _d⁴ ) δ 9.34(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 ₂₁ F ₂ N ₆ [M+H] ⁺ 395.2, found: 395.3.

Example 24

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

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

The synthetic route of compound 24 is shown in the figure above:

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 purified by silica gel column chromatography. ^¹H NMR (400 MHz, _CDCl₃ ): δ 7.20 (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).

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. The data for compound 24 obtained as in step d of the synthetic route in Example 1 are as follows: ^1H NMR (400MHz, MeOD-d ₄₎ )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 FN ₅ O[M+H] ⁺ 378.2,found:378.2.

Example 25

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

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

The synthetic route of compound 25 is shown in the figure above:

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 to the system 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 saline (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 (400 MHz, _CDCl3 ) δ 7.41 (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).

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. The data for compound 25 obtained as in step d of the synthetic route in Example 1 are as follows: ^1H NMR (400MHz, MeOD-d ₄ )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 FN ₅ O[M+H] ⁺ 392.2, found: 392.3.

Example 26

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

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

The synthetic route is shown in the figure above. 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,1H),7.56(dd,J＝ 7.2,1.2Hz,1H),7.50(d,J＝1.8Hz,1H),7.44(d,J＝2.4Hz,1H),7.34(t,J＝7.2 Hz,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 FN ₅ O[M+H] ⁺ 378.2, found:378.3.

Example 27

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

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

The synthetic route is shown in the figure above. 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 for C ₂₂ H ₂₃ FN ₅ O[M+H] ⁺ 392.2,found:392.3.

Example 28

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

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

The synthetic route of compound 28 is shown in the figure 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% MeCN in _H₂O containing 0.1% TFA over 90 min) to obtain compound 28 (2 mg, 20% yield). The data for compound 28 were: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 9.13 (s, 1H), 8.30 (s, 1H), 8.08–8.03 (m, 1H), 7.45 (d, J = 7.6 Hz, 1H), 7.35 (d, J = 7.6 Hz, 1H), 7.27 (t, J = 7.6 Hz, 1H), 7.17 (d, J = 8.4 Hz, 1H), 3.86 (s, 3H), 3.16–3.03 (m, 5H), 1.87–1.83 (m, 2H), 1.60 (brs, 2H) _. LRMS (ESI) calcd for _{C₂₀H₂₂FN₆O [} M+H] ^⁺ 381.2, found: 381.3.

Example 29

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

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

The synthetic route of compound 29 is shown in the figure above:

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 above system at 0 °C and stirred at room temperature for 4 h. After the reaction was complete, the solvent was removed by rotary evaporation under reduced pressure. The intermediate MC80 was obtained by silica gel column chromatography as a white oily liquid (7 mg, 71.6% yield). ^¹H NMR (400 MHz, _CDCl₃ ) δ 7.78 (dd, J = 8.0, 1.6 Hz, 1H), 7.55 (dd, J = 8.0, 1.6 Hz, 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 C₁₆H₁₇BrFN₄ [} M+H] _⁺ 363.1 ^, found: 363.1.

Step b, the same as synthesis step d in Example 1, yielded the following data for compound 29: ^¹H NMR (600MHz, MeOD-d ₄₎ )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 F ₂ N ₅ [M+H] ⁺ 380.2, found: 380.3.

Example 30

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)

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

The synthesis method of compound 30 is shown above:

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). ¹ H NMR (400MHz, CDCl ₃ )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).

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-d ₄ )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),2.4 3(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).

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 were as follows: ^¹H NMR (400MHz, MeOD- _d⁴ ) δ 9.09 (s, 1H), 8.27 (s, 1H), 8.03 (t, J = 8.0 Hz, 1H), 7.54 (d, J = 8.4 Hz, 1H), 7.41 (d, J = 8.4 Hz, 1H), 7.23 (dd, J = 8.4, 2.4 Hz, 1H), 3.88 (s, 3H), 3.16–3.14 (m, 2H), 2.47 (t, J = 7.2 Hz, 2H), 1.98–1.75 (m, 6H), 1.36–1.31 (m, 3H), 1.08 (t, J = 7.2 Hz, 3H). LRMS (ESI) [M+H] ^⁺ , found: 448.1.

Example 31

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

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

The synthetic route of compound 31 is shown in the figure above:

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 mixture was stirred at -78 °C for 2 h. After the reaction was complete, the mixture 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 using PE:EA (20:1-6:1) to obtain compound MC86 (1.4 g, 63.6% yield). ^1H NMR (400 MHz, _CDCl3 ) δ 7.35 (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).

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 _NaHCO3 aq (500 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 a white solid compound MC87 (600 mg, 37.4% yield). ^¹H NMR (400 MHz, _CDCl3 ) δ 7.94–7.90 (m, 1H), 7.88–7.83 (m, 1H), 7.27–7.22 (m, 1H), 3.25 (s, 3H).

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. ^¹H NMR (400 MHz, _CDCl₃ ) δ 8.07–8.04 (m, 2H), 7.77 (dd, J = 8.0, 2.4 Hz, 1H), 7.25 (t, J = 8.0 Hz, 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).

Step d: Using intermediate MC88, as in step d of the synthetic route in Example 1, the data for compound 31 are as follows: ^¹H NMR (400MHz, MeOD-d ₄₎ )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 FN ₅ O ₂ S[M+H] ⁺ 416.2,found:416.3.

Example 32

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

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

The synthesis route is shown in the figure above:

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. Then, it was washed with DCM and water, and the organic phase was concentrated under reduced pressure. After silica gel column chromatography and purification, 0.65 g of intermediate MC90 was obtained (yield 30%). ^¹H NMR (400 MHz, MeOD- _d⁴ ) δ 8.40 (s, 1H), 7.91 (d, J = 7.6 Hz, 1H), 7.66 (dd, J = 7.6, 2.4 Hz, 1H), 7.16 (t, J = 7.6 Hz, 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).

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 then purified by column chromatography to give 0.45 g of compound 32 (66% yield). ^¹H NMR (400 MHz, MeOD- _d⁴ ) δ 9.03 (s, 1H), 8.35 (d, J = 2.4 Hz, 1H), 8.12 (d, J = 8.0, 2.8 Hz, 1H), 7.71 (dd, J = 8.0, 1.6 Hz, 1H), 7.53 (dd, J = 7.6, 1.6 Hz, 1H), 7.37 (t, J = 8.0 Hz, 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 C₁₈H₂₀FN 6} O ₂ [M+H] ⁺ 383.2,found:383.2.

Example 33

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

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

The synthetic route of compound 33 is shown in the figure above:

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).

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) solution, and the system was heated to 60 °C and stirred for 2 hours. After the reaction was complete, 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). ^¹H NMR (400 MHz, MeOD- _d₄ ) δ 9.18–9.11 (m, 1H), 8.50 (s, 1H), 8.26–8.20 (m, 1H), 7.99–7.91 (m, 1H), 7.26–7.17 (m, 2H), 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 C₂₀H₂₂FN₆O [} M+H] ^⁺ 381.2, found: 381.2.

Example 34

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

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

The synthetic route of compound 34 is shown in the figure 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 while stirring. The mixture was heated to room temperature and the reaction continued for 2 hours. After the reaction was complete, 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. ^¹H NMR (400 MHz, MeOD- _d₄ ) δ 8.91 (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 for C₂¹H₂₄FN₆O [} M+H] ^⁺ 395.2, found: 395.3.

Example 35

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)

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)methanesul fonamide

The synthetic route is shown in the figure above. The synthetic method is the same as that for compound 33. The data for compound 35 are as follows: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 9.11 (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 for C₂₀H₂₁F₄N₆O₂S} [M+H _] ^⁺ _485.1 , found: 485.2.

Example 36

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

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

The synthetic route of compound 36 is shown in the figure above. The crude MC91 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 completed, the solvent was removed under reduced pressure. The product was separated and purified by prep-HPLC to obtain compound 36 as a yellow solid product (20 mg, yield 54%). ^¹H NMR (400 MHz, MeOD- _d₄ ) δ 9.12 (s, 1H), 8.22 (s, 1H), 7.97 (brs, 1H), 7.55–7.46 (m, 1H), 7.26 (t, J = 8.0 Hz, 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 for _{C₂₀H₂₄FN₆O 2} S[M+H] ⁺ 431.2,found:431.2.

Example 37

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

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

The synthesis method was the same as in Example 33, and the data for compound 37 were as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 8.98 (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 _{C₂₂H₂₆FN₆O₂S} [M+H] ^⁺ 457.2, found: 457.3.

Example 38

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

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33, yielding compound 38 as a white solid (8 mg, 17% yield). The structural characterization data are as follows: ^¹H NMR (600 MHz, MeOD- _d₄ ) δ 9.11 (s, 1H), 8.21 (s, 1H), 7.95 (brs, 1H), 7.56 (brs, 1H), 7.25 (t, J = 7.8 Hz, 1H), 7.19 (d, J = 7.8 Hz, 1H), 6.92 (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 _{C₂¹H₂₆FN₆O 2} S[M+H] ⁺ 445.2, found: 445.3.

Example 39

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 39 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 9.03(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 for _{C₂₂H₂₈FN₆O₂S [} M+ _H ] ^⁺ 459.2, found:459.3.

Example 40

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)

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)pyridine-2-sulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data of compound 40 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 9.15 (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 C₂₄H₂₅FN₇O₂S [ M} +H] ^⁺ 494.2, found: 494.3.

Example 41

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 (compound 41)

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 41 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 9.11 (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 _{C₂₃H₃₀FN₆O₂} S[M+H] ⁺ 473.2, found: 473.3.

Example 42

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)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 42 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 9.10 (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 C₂₃H₃₀FN₆O₂} S[M+H] ⁺ 473.2, found: 473.3.

Example 43

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)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 43 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 9.07-8.96 (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 for _{C₂₂H₂₈FN₆O₂} S[M+H] ⁺ 459.2, found: 459.3.

Example 44

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)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 44 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 9.02(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(brs, 1H), 3.83(s, 3H), 3.74(s, 2H), 3.46(brs, 2H), 3.10-2.97(m, 3H), 2.65(brs, 2H), 2.31(brs, 2H), 1.97(brs, 4H) ^. LRMS(ESI) calcd for _{C₂₂H₂₇ClFN₆O₂S} [ _M +H _{] ⁺} 493.2, found: 493.3.

Example 45

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)

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

The synthetic method is shown in the figure 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 were obtained as follows: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 9.11 (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 C₂¹H₂₆FN₆O₂S [ M +} H] ^⁺ 445.2, found: 445.3.

Example 46

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)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 46 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 9.02 (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 _{for C₂₃H₂₄FN₆O₂S₂ [} M+H _{]₂ +} ^499.1 , found: 499.3.

Example 47

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)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data of compound 47 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 9.09(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 C₂₀H₂₅FN₇O₂S [} M+H] ^⁺ _446.2 , found: 446.3.

Example 48

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)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 48 are as follows: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 9.16 (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 = 8.0Hz, 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 C₂₆H₂₈FN₆O₂S [} M+H] ⁺ _507.2 , found: 507.2.

Example 49

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)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 49 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 9.15(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) calcd _{for C₂₂H₂₈FN₆O₃S [} M+H] _⁺ 475.2, found:475.2.

Example 55

N-(3-(5-(cyclopropylmethoxy)-6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide (compound 55)

N-(3-(5-(cyclopropylmethoxy)-6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide

The synthetic route is shown in the figure above: Step a, compound 90 (365 mg, 1.0 mmol, 1.0 eq) and reduced iron powder (560 mg, 10.0 mmol, 10.0 eq) were dissolved in a mixed solvent of EtOH (20 mL) and H₂O ( ₂ mL), and then _NH₄Cl (560 g, 10.0 mmol, 10.0 eq) was added. The reaction system was heated to 50 °C and stirred for 3 h. After the reaction was completed, DCM (200 mL) was added to dilute the system, and the mixture was filtered and concentrated under reduced pressure to obtain crude product (500 mg). The crude product (33 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 completed, the solvent was removed under reduced pressure, and the product was separated and purified by silica gel chromatography to obtain compound 92 as a yellow solid product (20 mg, yield 54%). ^¹H NMR (400 MHz, MeOD- _d₄ ) δ 8.58 (s, 1H), 8.41 (s, 1H), 8.07 (s, 1H), 3.68 (s, 3H), 3.64–3.57 (m, 2H), 3.29–3.14 (m, 5H), 2.94–2.88 (m, 1H), 2.23–2.13 (m, 2H), 2.04–1.99 (m, 2H).

In step b, intermediates N-(3-bromo-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide (0.735 g, 1.78 mmol, 1.0 eq), _Na₂CO₃ ( _0.38 g, 3.56 mmol, 2.0 eq) and 3-(cyclopropylmethoxy)-2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxabor-2-yl)pyridine (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 0.587 g (66% yield). ^¹H NMR (400 MHz, MeOD-d₄ ₎ )δ9.08(s,1H),7.76-7.68(m,2H),7.55-7.46(m,1H),7.26(t,J＝8.0Hz,1H),7.20-7.17(m,1H),3.98(s,3H),3.93(d,J＝7.2Hz,1H),3.8 6(s,3H),3.10-2.99(m,3H),2.69-2.64(m,2H),2.04-1.94(m,4H),1.28(brs,4H)0.71-0.66(m,2H),0.41-0.36(m,2H).LRMS(ESI)calcd for C ₂₄ H ₃₀ FN ₆ O ₃ S[M+H] ⁺ 500.2,found:501.3.

Example 57

N-(3-(2-(tert-butyl)-2H-pyrazolo[3,4-b]pyridin-5-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide (compound 57)

N-(3-(2-(tert-butyl)-2H-pyrazolo[3,4-b]pyridin-5-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 55. The data for compound 57 are as follows: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 9.13 (s, 1H), 8.98 (s, 1H), 8.51 (brs, 1H), 8.04 (s, 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), 3.10-2.97 (m, 3H), 2.68-2.65 (m, 2H), 2.05-1.95 (m, 4H), _1.20 (s, 9H) _. LRMS ₍ ESI) calcd for _{C₂₅H₃₃FN₈O₂S} [M+H] ^⁺ 508.2, found: 509.3.

Example 65

N-(2-(4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)phenyl)butane-1-sulfonamide (compound 65)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 65 are: LRMS(ESI)calcd for C ₂₃ H ₂₉ F ₂ N ₆ O ₂ S[M+H] ⁺ 491.2,found:491.3.

Example 67

3-Chloro-N-(2-(4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)phenyl)propane-1-sulfonamide (Compound 67)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 67 are: LRMS(ESI)calcd for C ₂₂ H ₂₆ ClF ₂ N ₆ O ₂ S[M+H] ⁺ 511.1,found:511.2.

Example 68

N-(2-(4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)phenyl)thiophene-2-sulfonamide (compound 68)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 68 are: LRMS(ESI)calcd for C ₂₃ H ₂₃ ClF ₂ N ₆ O ₂ S ₂ [M+H] ⁺ 517.1,found:517.2.

Example 75

3,3,3-Trifluoro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)propane-1-sulfonamide (Compound 75)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 75 are: LRMS(ESI)calcd for C ₂₂ H ₂₅ F ₄ N ₆ O ₂ S[M+H] ⁺ 513.2,found:513.3.

Example 76

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)tetrahydro-2H-pyran-4-sulfonamide (compound 76)

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)tetrahydro-2H-pyran-4-sulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 76 are: LRMS(ESI)calcd for C ₂₄ H ₃₀ FN ₆ O ₃ S[M+H] ⁺ 501.2,found:501.4.

Example 77

4-(N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)sulfanilyl)methyl benzoate (compound 77)

methyl

4-(N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)sulfamoyl)benzoate

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 77 are as follows: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 9.00 (s, 1H), 8.20-8.17 (m, 6H), 7.53 (brs, 1H), 7.16-7.13 (m, 3H), 3.94 (s, 3H), 3.82 (s, 3H), 3.15-2.51 (m, 5H), 1.89-1.86 (m, 4H) _. LRMS (ESI) calcd _{for C₂₇H₂₈FN₆O₄S [} M+H] ⁺ _551.2 , found: 551.2.

Example 78

4-Fluoro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)benzenesulfonamide (Compound 78)

4-fluoro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)benzenesulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data of compound 78 are as follows: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 9.03 (s, 1H), 8.12-7.91 (m, 4H), 7.55 (brs, 1H), 7.32-7.29 (m, 2H), 7.19-7.14 (m, 3H), 3.83 (s, 3H), 3.03-2.52 (m, 5H), _2.03-1.30 (m, 4H) _. LRMS ( _{ESI )} calcd for _{C₂₅H₂₅F₂N₆O₂S} [M+H] ⁺ 511.2, found: 511.2.

Example 79

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-6-methylpyridin-3-sulfonamide (compound 79)

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-6-methylpyridine-3-sulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data of compound 79 are as follows: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 9.15 (s, 1H), 8.81 (brs, 1H), 8.24-8.12 (m, 2H), 7.92 (brs, 1H), 7.53-7.35 (m, 1H), 7.22-7.15 (m, 4H), 3.87 (s, 3H), 3.15-2.56 (m, 8H), 2.03-1.59 (m, 4H). _{LRMS (ESI) calcd for C₂₅H₂₇FN₇O₂S [ M} +H] ⁺ _508.2 , found: 508.2.

Example 80

2-(4-fluoro-4-(4-(methyl-d3)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile (Compound 80)

2-(4-fluoro-4-(4-(methyl-d3)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data for compound 80 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 8.76 (s, 1H), 8.28 (d, J = 2.4Hz, 1H), 8.06 (td, J = 7.8, 2.4Hz, 1H), 7.75 (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), 3.41 (brs, 2H), 3.22-3.14 (m, 2H), 2.30-2.23 (m, 4H) _. LRMS(ESI) calcd for _{C₂₀H₁₆D₃F₂N₆} [ _M +H ^] _⁺ 384.2, found:384.2.

Example 81

N-(2-(4-fluoro-4-(4-(methyl-d3)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)phenyl)methanesulfonamide (compound 81)

N-(2-(4-fluoro-4-(4-(methyl-d3)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)phenyl)methanesulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data of compound 81 are as follows: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 8.53 (s, 1H), 8.20 (d, J = 1.6Hz, 1H), 7.97 (brs, 1H), 7.55 (brs, 1H), 7.26 (t, J = 5.2Hz, 1H), 7.17 (brs, 1H), 6.98 (brs, 1H), 3.17 (s, 3H), 3.06-2.70 (m, 4H), 2.40 (brs, 2H), 2.22-2.17 (m _{, 2H )} . LRMS ( _ESI ) calcd for _{C₂₀H₂₀F₂N₆O₂S} [M+H] ⁺ 452.2, found: 452.2.

Example 83

3-(6-Fluoropyridin-3-yl)-2-(4-(4-(methyl-d3)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile (Compound 83)

3-(6-fluoropyridin-3-yl)-2-(4-(4-(methyl-d3)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data for compound 83 are as follows: ^¹H NMR (400MHz, MeOD- _d4 ) δ 9.01 (s, 1H), 8.26 (d, J = 2.4Hz, 1H), 8.02 (td, J = 8.4, 2.4Hz, 1H), 7.73 (dd, J = 8.0, 1.6Hz, 1H), 7.56 (dd, J = 8.0, 1.6Hz, 1H), 7.33 (t, J = 8.0Hz, 1H), 7.20 (dd, J = 8.4, 2.4Hz, 1H), 3.27 (brs, 2H), 3.17-3.08 (m, 3H), 1.97-1.93 (m, 2H), 1.83-1.79 (m, 2H). ^¹³C NMR (150MHz, MeOD- _d4) )δ164.4(d,J＝238.4Hz),159.1,154.2,148.5(d,J＝14.3Hz),145.8,144.1(d,J＝8.3Hz),137.6,136.9,135.9,1 35.1(d,J＝2.7Hz),126.0,119.5,111.2,110.5(d,J＝37.1Hz),52.5,32.8,32.6-31.9(m),30.8.HRMS(ESI)calcd for C ₂₀ H ₁₇ D ₃ FN ₆ [M+H] ⁺ :366.1916,found:366.1918.

Example 84

3-(5,6-Difluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile (Compound 84)

3-(5,6-difluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data of compound 84 are as follows: ^¹H NMR (400MHz, MeOD) δ 9.02 (s, 1H), 8.05 (s, 1H), 8.01-7.89 (m, 1H), 7.75 (dd, J = 7.6, 1.6Hz, 1H), 7.58 (dd, J = 8.0, 2.0Hz, 1H), 7.34 (t, J = 8.0Hz, 1H), 3.85 (s, 3H), 3.25-3.07 (m, 4H), 1.97 (d, J = 12.4Hz, 2H), 1.89-1.72 (m, 2H). LRMS (ESI) calcd for C ₂₀ H ₁₉ F ₂ N ₆ [M+H] ⁺ 381.2, found: 381.3.

Example 85

3-(6-chloro-5-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile (Compound 85)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data of compound 85 are as follows: ^1H NMR (600MHz, MeOD) δ 9.00 (s, 1H), 8.23-8.10 (m, 2H), 7.75 (d, J = 7.2Hz, 1H), 7.59 (d, J = 7.2Hz, 1H), 7.35 (t, J = 7.8Hz, 1H), 3.85 (s, 3H), 3.26-3.05 (m, 4H), 2.03-1.90 (m, 2H), 1.80 (s, 2H). LRMS (ESI) calcd for C ₂₀ H ₁₉ FClN ₆ [M+H] ⁺ 397.1, found: 397.2.

Example 86

3-(6-fluoro-5-methoxypyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile (Compound 86)

3-(6-fluoro-5-methoxypyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data of compound 86 are as follows: ^1H NMR (400MHz, MeOD) δ 9.08 (s, 1H), 7.76-7.68 (m, 2H), 7.63 (dd, J = 9.6, 2.0Hz, 1H), 7.56 (dd, J = 7.6, 1.6Hz, 1H), 7.31 (t, J = 7.6Hz, 1H), 3.98 (s, 3H), 3.86 (s, 3H), 3.36 (s, 2H), 3.22-3.07 (m, 2H), 1.97 (d, J = 11.6Hz, 2H), 1.92-1.76 (m, 2H). LRMS (ESI) calcd for C ₂₁ H ₂₂ FN ₆ O [M+H] ⁺ 393.2, found: 393.2.

Example 87

3-(6-fluoro-5-methylpyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile (Compound 87)

3-(6-fluoro-5-methylpyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data of compound 87 are as follows: ^1H NMR (600MHz, MeOD) δ 9.02 (s, 1H), 8.07 (s, 1H), 7.88 (d, J = 9.6Hz, 1H), 7.73 (dd, J = 7.8, 1.8Hz, 1H), 7.59-7.54 (m, 1H), 7.34 (t, J = 7.8Hz, 1H), 3.86 (s, 3H), 3.28-3.06 (m, 4H), 2.39 (s, 3H), 1.96 (d, J = 12.6Hz, 2H), 1.83 (s, 2H). LRMS (ESI) calcd for C ₂₁ H ₂₂ FN ₆ [M+H] ⁺ 377.3, found: 377.4.

Example 88

3-(6-fluoro-4-methylpyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile (Compound 88)

3-(6-fluoro-4-methylpyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1, and the data for compound 88 are as follows: ¹ H NMR (400MHz, MeOD) δ9.08 (s, 1H), 7.99 (s, 1H), 7.75 (dd, J = 8.0, 2.0Hz, 1H), 7.45 ( dd,J＝7.6,1.6Hz,1H),7.34(t,J＝8.0Hz,1H),7.11(s,1H),3.84(s,3H),3.43-3.3 4(m,2H),3.22-3.05(m,2H),2.99-2.86(m,1H),2.25(s,3H),1.97(d,J＝13.2Hz,1 H),1.88(d,J＝13.2Hz,1H),1.81-1.69(m,1H),1.70-1.46(m,1H).LRMS(ESI)calcd for C ₂₁ H ₂₂ FN ₆ [M+H] ⁺ 377.3,found:377.3.

Example 89

N-(3-cyano-5-(6-fluoropyridin-3-yl)-4-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-1-methylpiperidin-4-carboxamide (Compound 89)

N-(3-cyano-5-(6-fluoropyridin-3-yl)-4-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-1-methylpiperidine-4-carboxamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data for compound 89 are: LRMS(ESI)calcd for C ₂₇ H ₃₂ FN ₈ O[M+H] ⁺ 503.2,found:503.4.

Example 90

3-(6-Fluoropyridin-3-yl)-2-(4-(4-(4-methoxybenzyl)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile (Compound 90)

3-(6-fluoropyridin-3-yl)-2-(4-(4-(4-methoxybenzyl)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)benzonitrile

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 1. The data for compound 90 are as follows: ^¹H NMR (600MHz, _CDCl₃ ) δ 8.47(s, 1H), 8.16(s, 1H), 7.92(t, J = 8.4Hz, 1H), 7.63(d, J = 8.4Hz, 1H), 7.43(d, J = 7.2Hz, 1H), 7.25-7.23(m, 1H), 7.11-7.09(m, 3H), 6.93(d, J = 7.2Hz, 2H), 5.10(s, 2H), 3.82(s, 3H), 3.22-3.12(m, 4H), 2.83(t, J = 12.0Hz, 1H), 1.86(brs, 2H), 1.73-1.70(m, 2H). ¹³ C NMR(125MHz,MeOD- _d4 )δ164.4(d,J＝238.3Hz),161.9,159.1,154.2,148.5(d,J＝14.1Hz),145.1,144.1(d,J＝8.0Hz),137.6,136.9,135.9,135.1( d,J＝3.3Hz),131.0,126.4,126.0,119.4,115.8,111.2,110.5(d,J＝36.9Hz),55.9,52.6,50.2,33.3,31.3.HRMS(ESI)calcd for C ₂₇ H ₂₆ FN ₆ O[M+H] ⁺ :469.2147,found:469.2149.

Example 91

2-(4-(4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)benzonitrile (Compound 91)

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

The synthetic route is shown in the figure above. Compound 90 (24 mg, 0.05 mmol, 1.0 equiv) was dissolved in TFA (2 mL), and the reaction was heated to 50 °C and stirred for 4 h. After cooling to room temperature, the solvent was removed by rotary evaporation. The crude product was purified by preparative liquid chromatography, and the data for compound 91 were as follows: ^¹H NMR (600 MHz, MeOD- _d4) )δ8.53(brs,1H),8.25(s,1H),8.02(t,J＝7.8Hz,1H),7.72(d,J＝7.8Hz,1H),7.55(d,J＝7.8Hz,1H),7.32(t,J＝7.8Hz,1 H),7.21(d,J＝8.4Hz,1H),3.27-3.24(m,2H),3.13(brs,2H),2.98-2.93(m,1H),1.98-1.94(m,2H),1.81-1.74(m,2H). ^13C NMR(125MHz,MeOD- _d4 )δ164.4(d,J＝238.4Hz),161.5,154.3,148.4(d,J＝14.1Hz),146.0,144.2(d,J＝8.3Hz),137.6,136.8,1 35.9,135.1(d,J＝5.3Hz),125.9,119.5,111.1,110.4(d,J＝36.9Hz),52.6,34.8,31.5.HRMS(ESI)calcd for C ₁₉ H ₁₈ FN ₆ [M+H] ⁺ :349.1571,found:349.1572.

Example 92

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-(methyl-d3)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide (compound 92)

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-(methyl-d3)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 92 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 8.92(s,1H), 8.22(s,1H), 7.96(s,1H), 7.56(s,1H), 7.26(t,J＝7.9Hz,1H), 7.19(d,J＝7.7Hz,1H), 6.94(s,1H), 3.17(s,3H), 3.07(m,2H), 2.96(m,1H), 2.63(m, _2H ), _1.94 (m, _4H ). _{LRMS(ESI) calcd for C₂₀H₂₁D₃FN₆O₂S [} M+H] ^⁺ 434.2, found: 434.2.

Example 93

N-(2-(4-fluoro-4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)phenyl)methanesulfonamide (compound 93)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 93 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 8.43(s, 1H), 8.20(s, 1H), 7.97(s, 1H), 7.55(brs, 1H), 7.26(t, J = 7.8Hz, 1H), 7.18-7.16(m, 1H), 6.98(s, 1H), 3.81(s, 3H), 3.17(s, 3H), 2.92(brs, _{4H), 2.40(brs, 2H), 2.22-1.91(m, 2H). LRMS (ESI) calcd for C₂₀H₂₄FN₆O₂S [ M} +H] ^⁺ _449.2 , found: 449.2.

Example 94

N-(6'-fluoro-4-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-[3,3'-bipyridine]-5-yl)methanesulfonamide (compound 94)

N-(6'-fluoro-4-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-[3,3'-bipyridin]-5-yl)methanesulfonamide

The synthetic route of compound 94 is shown in the figure above:

In step a, MC53 (500 mg, 3.01 mmol, 1 eq), MC104 (1.07 g, 4.52 mmol, 1.5 eq), and potassium carbonate (541 mg, 3.91 mmol) were dissolved in DMSO (5 mL), and the reaction was carried out at 50 °C for 2 h. The reaction was quenched with water and then extracted with EA and water. The organic phase was distilled under reduced pressure, and the concentrate was purified by silica gel column chromatography using DCM:MeOH (40:1-10:1) to give a pale yellow oily liquid compound MC105 (580 mg, 53% yield). ^¹H NMR (600 MHz, CDCl₃) δ 8.73 (s, 2H), 8.07 (s, 1H), 3.68 (s, 3H), 3.46 (m, 2H), 3.16 (t, J = 13.6 Hz, 2H), 2.95 (tt, J = 11.7, 3.7 Hz, 1H), 2.31 (qd, J = 11.9, 4.0 Hz, 2H), 2.01 (m, 2H).

In step b, MC105 (142 mg, 0.39 mmol, 1 eq), Fe (219 mg, 3.9 mmol, 10 eq), and _NH4Cl (211 mg, 3.9 mmol, 10 eq) were dissolved in ethanol (5 mL) and water (1 mL). The temperature was raised to 70 °C, and the mixture was stirred for 2 h. After the reaction was complete, the mixture was filtered while hot, concentrated under reduced pressure, and the concentrate was dissolved in DCM and filtered to remove salt. The filtrate was concentrated under reduced pressure to obtain a grayish-white solid, crude MC106.

In step c, MC106 (131 mg, 0.39 mmol, 1 eq), methanesulfonyl chloride (89 mg, 0.78 mmol, 2 eq) and pyridine (62 mg, 0.78 mmol, 2 eq) were dissolved in DCM (5 mL), the temperature was raised to 50 °C, and the reaction was stirred for 12 h. The solution was quenched with water, extracted with EA and water, concentrated under reduced pressure, and purified by silica gel column chromatography with DCM:MeOH (40:1-10:1) to give a brown oily liquid compound MC107 (138 mg, 88%). ^¹H NMR (400 MHz, CDCl₃) δ 8.58 (s, 1H), 8.41 (s, 1H), 8.07 (s, 1H), 3.68 (s, 3H), 3.57 (m, 2H), 3.14 (s, 3H), 2.91 (m, 1H), 2.18 (m, 2H), 2.01 (m, 2H), 1.83 (m, 2H).

In step d, MC108 (162 mg, 0.39 mmol, 1 eq), 2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)pyridine (132 mg, 0.59 mmol, 1.5 eq), and sodium carbonate (100 mg, 0.78 mmol, 2 eq) were dissolved in 1,4-dioxane (5 mL) and water (0.5 mL). Pd(dppf) _Cl₂ was added under argon protection. The temperature was raised to 85 °C, and the reaction was stirred for 12 h. The reaction solution was concentrated under reduced pressure, and the concentrate was separated and purified by prep-HPLC to obtain a yellow solid compound 94. The data for compound 94 are as follows: ^¹H NMR (600MHz, MeOD) δ 8.99 (s, 1H), 8.56 (s, 1H), 8.39 (d, J = 2.8Hz, 1H), 8.31 (s, 1H), 8.13 (td, J = 7.6, 2.7Hz, 1H), 7.29 (dd, J = 8.4, 2.6Hz, 1H), 3.82 (s, 3H), 3.57 (m, 2H), 3.26 (m, 2H), 3.21 (s, 1H), 3.19 (s, 3H), 1.92 (m, 2H), 1.84 (m, 2H). LRMS (ESI) calcd for C ₁₉ H ₂₃ FN ₆ O ₂ S [M+H] ⁺ 432.2, found: 432.2.

Example 95

N-(5-fluoro-3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide (compound 95)

N-(5-fluoro-3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 95 are: LRMS(ESI)calcd for C ₂₀ H ₂₃ F ₂ N ₆ O ₂ S[M+H] ⁺ 449.1,found:449.2.

Example 96

N-(5-chloro-3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide (compound 96)

N-(5-chloro-3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 96 are: LRMS(ESI)calcd for C ₂₀ H ₂₃ ClFN ₆ O ₂ S[M+H] ⁺ 465.1, found: 465.2.

Example 97

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-5-(trifluoromethyl)phenyl)methanesulfonamide (compound 97)

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-5-(trifluoromethyl)phenyl)methanesulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data of compound 97 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 9.08 (s, 1H), 8.27 (d, J = 2.4Hz, 1H), 8.01 (brs, 1H), 7.76 (brs, 1H), 7.33 (brs, 1H), 7.22 (dd, J = 8.4, 2.4Hz, 1H), 3.84 ₍ s, 3H), 3.19 (s, 3H), 3.14-2.52 (m, 5H), 2.04-1.92 (m, _4H ) _. LRMS (ESI) calcd for _{C₂¹H₂³F₄N₆O₂S} [M+H] ⁺ _499.1 , found: 499.2.

Example 98

N-(3-(6-fluoropyridin-3-yl)-5-methoxy-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide (compound 98)

N-(3-(6-fluoropyridin-3-yl)-5-methoxy-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methanesulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 98 are: LRMS(ESI)calcd for C ₂₁ H ₂₆ FN ₆ O ₃ S[M+H] ⁺ 461.2,found:461.2.

Example 99

N-(3-(6-fluoropyridin-3-yl)-2-((1R,5S)-3-(4-methyl-4H-1,2,4-triazol-3-yl)-8-azabicyclo[3.2.1]octane-8-yl)phenyl)methanesulfonamide (compound 99)

N-(3-(6-fluoropyridin-3-yl)-2-((1R,5S)-3-(4-methyl-4H-1,2,4-triazol-3-yl)-8-azabicyclo[3.2.1]octan-8-yl)phenyl)methanesulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data of compound 99 are as follows: ^¹H NMR (600MHz, MeOD- _d₄ ) δ 9.10 (s, 1H), 8.23 (s, 1H), 7.40-7.38 (m, 1H), 7.13-7.07 (m, 3H), 3.99 (brs, 2H), 3.93 (s, 3H), 3.52-3.45 (m, 1H), 3.18 (s, 3H), 1.90 (brs, 2H), 1.78-1.73 (m, 6H) _{. LRMS (ESI) calcd for C₂₂H₂₆FN₆O₂S [} M+H] _{+ 457.2} , found ^: 457.2.

Example 100

N-(2-(4-(5-amino-4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)phenyl)methanesulfonamide (compound 100)

N-(2-(4-(5-amino-4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-(6-fluoropyridin-3-yl)phenyl)methanesulfonamide

The synthetic route for compound 100 is shown in the figure above.

Step a: Compound MC89 (1.18 g, 5.0 mmol, 1.0 eq), compound MC1 (1.43 g, 10.0 mmol, 2.0 eq), and _K3PO4 (2.12 g, 10.0 mmol, 2.0 eq) were dissolved in a mixed solvent of DMSO (10 mL) _. The system was heated to 80 °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 MC2 (1.6 g, 93.3% yield). ¹ H NMR (400MHz, CDCl ₃ )δ7.78(d,J＝8.0Hz,1H),7.58(d,J＝8.0Hz,1H),7.04(t,J＝8.0Hz,1H),3.73(s,3H),3.18-2.96(m,4H),2.51-2.47(m,1H),1.97-1.93(m,4H).

In step b, compound MC2 (1.6 g, 4.66 mmol, 1.0 eq) and reduced iron powder (2.8 g, 10.0 mmol, 50.0 eq) were dissolved in a mixed solvent of EtOH (20 mL) and _H₂O (2 mL), and then _NH₄Cl (2.8 g, 10.0 mmol, 50.0 eq) was added. The reaction system was heated to 50 °C and stirred for 3 h. After the reaction was completed, DCM (200 mL) was added to dilute the system, and the mixture was filtered and concentrated under reduced pressure to obtain the crude product MC3 (2.0 g). ¹ H NMR (400MHz, CDCl ₃ )δ6.81-6.78(m,2H),6.66(dd,J＝6.4,2.8Hz,1H),4.36(brs,3H),3.71(s,3H),3.59-3. 52(m,2H),2.91-2.85(m,2H),2.48-2.39(m,1H),2.03-1.99(m,2H),1.81-1.70(m,2H).

Step c: Dissolve crude product MC3 (2.0 g, crude) in DCM (20.0 mL), and add methanesulfonyl chloride (773 mg, 6.75 mmol, 1.5 eq) and pyridine (712 mg, 9 mmol, 2.0 eq) at 0 °C. The reaction was heated to 50℃ and stirred for 2 hours. After the reaction was completed, the solvent was removed under reduced pressure, and the product was separated and purified by silica gel chromatography to obtain compound MC4 as a yellow solid product (1.76 g, yield 99%). ^¹H NMR (400 MHz, MeOD- _d₄ ) δ 7.50 (d, J = 8.0 Hz, 1H), 7.22 (d, J = 8.0 Hz, 1H), 7.06 (t, J = 8.0 Hz, 1H), 3.72–3.63 (m, 5H), 3.03 (s, 3H), 2.83–2.79 (m, 2H), 2.48–2.40 (m, 1H), 2.12–2.06 (m, 2H), 1.84–1.73 (m, 2H).

Step d: Dissolve crude product MC4 (1.76 g, 4.5 mmol, 1.0 eq) in EtOH (20.0 mL) and add hydrazine hydrate (3 mL). The reaction was heated to 90℃ and stirred for 16 hours. After the reaction was completed, the solvent was removed under reduced pressure, and the product was separated and purified by silica gel chromatography to obtain compound MC5 as a white solid product (1.24 g, yield 71%). ^¹H NMR (400MHz, DMSO- _d⁶ ) δ 9.02 (brs, 1H), 8.48 (brs, 1H), 7.44 (d, J = 8.0 Hz, 1H), 7.29 (d, J = 7.2 Hz, 1H), 7.15 (t, J = 8.0 Hz, 1H), 4.19 (brs, 2H), 3.52–3.45 (m, 2H), 3.17 (s, 3H), 2.79–2.75 (m, 2H), 2.22–2.15 (m, 1H), 1.87–1.77 (m, 2H), 1.72–1.61 (m, 2H).

Step e: Dissolve MC5 (610 mg, 1.56 mmol, 1.0 eq) in dioxane (10 mL) and water (1 mL), and add CNBr (497 mg, 4.69 mmol, 3.0 eq) and _NaHCO3 (524 mg, 6.24 mmol, 4.0 eq). After stirring at room temperature for 2 hours, the reaction mixture was quenched with 10% NaHCO3 solution ( ₂₀ mL). The precipitated solid was obtained by filtration and washed with water (20 mL). The resulting solid was further dried under vacuum to give MC6 (530 mg, 78.8%) as a pale yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ )δ7.45(d,J＝8.0Hz,1H),7.33(d,J＝7.6Hz,1H),7.17(t,J＝8.0Hz,1H),3. 62-3.56(m,2H),3.22-3.18(m,4H),2.89-2.85(m,2H),2.05-1.86(m,4H).

Step f: Add N-dimethylformamide dimethyl acetal (0.33 mL, 2.5 mmol, 2.0 eq) to a solution of MC6 (530 mg, 1.23 mmol, 1.0 eq) in dioxane (4 mL) at room temperature. Stir the reaction mixture at 100 °C for 2 h. After complete consumption of the starting material as monitored by TLC, concentrate the mixture under reduced pressure to obtain MC7 (600 mg, crude) as a yellow solid. The crude product itself is used in the next step without any further purification.

Step g: Add a solution of 600 mg (crude) of dioxane (6 mL) to a sealing tube, and add methylamine (25% MeOH solution, 6 mL) and AcOH (0.220 mL). Stir the reaction mixture at 110 °C for 16 hours. After the reaction is complete, remove the solvent under reduced pressure, and purify the product by silica gel chromatography to obtain compound MC8 as a white solid product (230 mg, two-step yield 43.6%). LRMS(ESI)calcd for C ₁₅ H ₂₂ BrN ₆ O ₂ S[M+H] ⁺ 429.2,found:429.1.

In step h, intermediate MC8 (85 mg, 0.20 mmol, 1.0 eq), sodium carbonate (64 mg, 0.6 mmol, 3.0 eq), and 4-fluoro-3-pyridineboronic acid (89 mg, 0.4 mmol, 2.0 eq) were dissolved in a mixed solvent of 4 mL dioxane and 0.4 mL water. Under nitrogen protection, Pd(dppf) _Cl₂ (15 mg, 0.02 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 100 were: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 8.95 (s, 1H), 8.63–8.29 (m, 2H), 7.58–7.42 (m, 2H), 7.30 (t, J = 7.4Hz, 1H), 7.01 (brs, 1H), 3.78 (s, 3H), 3.17–3.10 (m, 6H), 2.92–2.68 (m, 2H), 1.96–1.85 (m, 4H). LRMS (ESI) calcd for _{C₂₀H₂₅FN₇O₂S [ M} + _H ] ^⁺ 446.2, found: 446.2.

Example 101

N-(2-(4-(5-amino-4-methyl-4H-1,2,4-triazol-3-yl)-4-fluoropiperidin-1-yl)-3-(6-fluoropyridin-3-yl)phenyl)methanesulfonamide (chemical)

Compound 101)

N-(2-(4-(5-amino-4-methyl-4H-1,2,4-triazol-3-yl)-4-fluoropiperidin-1-yl)-3-(6-fluoropyridin-3-yl)phenyl)methanesulfonamide

The synthetic route for compound 101 is shown in the figure above. The data for compound 101 are: LRMS(ESI)calcd for C ₂₀ H ₂₄ F ₂ N ₇ O ₂ S[M+H] ⁺ 464.2,found:464.2.

Example 102

N-(2-(4-(5-amino-4-methyl-4H-1,2,4-triazol-3-yl)-4-methylpiperidin-1-yl)-3-(6-fluoropyridin-3-yl)phenyl)methanesulfonamide (compound 102)

N-(2-(4-(5-amino-4-methyl-4H-1,2,4-triazol-3-yl)-4-methylpiperidin-1-yl)-3-(6-fluoropyridin-3-yl)phenyl)methanesulfonamide

The synthetic route for compound 102 is shown in the figure above. The data for compound 102 are: LRMS(ESI)calcd for C ₂₁ H ₂₇ FN ₇ O ₂ S[M+H] ⁺ 460.2,found:460.3.

Example 103

2-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)isothiazolidin-1,1-dioxide (compound 103)

2-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)isothiazolidine 1,1-dioxide

The synthetic route for compound 103 is shown in the figure above. The data for compound 103 are: LRMS(ESI)calcd for C ₂₂ H ₂₆ FN ₆ O ₂ S[M+H] ⁺ 457.2,found:457.3.

Example 106

3-Fluoro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)propane-1-sulfonamide (Compound 106)

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

The synthetic route for compound 106 is shown in the figure above. The data for compound 106 are: LRMS(ESI)calcd for C ₂₂ H ₂₇ F ₂ N ₆ O ₂ S[M+H] ⁺ 477.2,found:477.2.

Example 107

4-Chloro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)butane-1-sulfonamide (Compound 107)

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

The synthetic route for compound 107 is shown in the figure above. The data for compound 107 are: LRMS(ESI)calcd for C ₂₃ H ₂₉ FClN ₆ O ₂ S[M+H] ⁺ 507.2,found:507.2.

Example 108

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-3-methylbutane-1-sulfonamide (compound 108)

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-3-methylbutane-1-sulfonamide

The synthetic route for compound 108 is shown in the figure above. The data for compound 108 are: LRMS(ESI)calcd for C ₂₄ H ₃₂ FN ₆ O ₂ S[M+H] ⁺ 487.2,found:487.3.

Example 109

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)benzenesulfonamide (compound 109)

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)benzenesulfonamide

The synthetic route for compound 109 is shown in the figure above. The data for compound 109 are: LRMS(ESI)calcd for C ₂₅ H ₂₆ FN ₆ O ₂ S[M+H] ⁺ 493.2,found:493.2.

Example 110

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)pentane-1-sulfonamide (compound 110)

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)pentane-1-sulfonamide

The synthetic route for compound 110 is shown in the figure above. The data for compound 110 are: LRMS(ESI)calcd for C ₂₄ H ₃₂ FN ₆ O ₂ S[M+H] ⁺ 487.2,found:487.3.

Example 111

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)hexane-1-sulfonamide (compound 111)

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)hexane-1-sulfonamide

The synthetic route for compound 111 is shown in the figure above. The data for compound 111 are: LRMS(ESI)calcd for C ₂₅ H ₃₄ FN ₆ O ₂ S[M+H] ⁺ 501.2,found:501.3.

Example 112

4-Chloro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)benzenesulfonamide (compound 112)

4-chloro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)benzenesulfonamide

The synthetic route for compound 112 is shown in the figure above. The data for compound 112 are: LRMS(ESI)calcd for C ₂₅ H ₂₅ ClFN ₆ O ₂ S[M+H] ⁺ 527.1,found:526.8.

Example 113

4-Chloro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)butane-1-sulfonamide (compound 113)

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

The synthetic route for compound 113 is shown in the figure above. The data for compound 113 are: LRMS(ESI)calcd for C ₂₃ H ₂₉ ClFN ₆ O ₂ S[M+H] ⁺ 507.2,found:507.2.

Example 114

3-Chloro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)benzenesulfonamide (compound 114)

3-chloro-N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)benzenesulfonamide

The synthetic route for compound 114 is shown in the figure above. The data for compound 114 are: LRMS(ESI)calcd for C ₂₅ H ₂₅ ClFN ₆ O ₂ S[M+H] ⁺ 527.1,found:527.2.

Example 115

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-3-hydroxypropane-1-sulfonamide (compound 115)

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-3-hydroxypropane-1-sulfonamide

The synthetic route of compound 115 is shown in the figure above:

Step a: 3-Bromopropanol MC123 (1 g, 7.2 mmol, 1 eq) and DMAP were dissolved in pyridine (15 mL). After the system temperature was lowered to 0 °C, p-methyl tert-butyryl chloride (955 mg, 7.92 mmol, 1.1 eq) was added dropwise, and then the temperature was raised to room temperature and the reaction was stirred for 30 min. The reaction was quenched with saturated _NH4Cl aqueous solution, 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 to obtain a colorless oily liquid compound MC124 (1.38 g, 86% yield). ^¹H NMR (600 MHz, CDCl3) δ 4.19 (t, J = 6.0 Hz, 2H), 3.46 (t, J = 6.6 Hz, 2H), 2.18 (p, J = 6.3 Hz, 2H), 1.20 (s, 9H).

Step b: MC124 (1.38 g, 6.2 mmol, 1 eq) and thiourea (567 mg, 7.44 mmol, 1.2 eq) were dissolved in ethanol (7 ml). The mixture was heated to 80 °C and stirred for 2 h. The solvent was then removed by concentration under reduced pressure. The concentrate was dissolved in DCM, filtered to remove thiourea, and the solvent was removed by rotary evaporation under reduced pressure. The concentrate was then washed with diethyl ether to remove excess MC125, and the filtered product was a white solid crude product MC125 (1.33 g, 98% yield).

Step c: Crude MC125 was dissolved in water (0.9 mL) and acetonitrile (15 mL), the temperature was lowered to 0 °C, tert-butyl hypochlorite was added dropwise, and the reaction was stirred for 30 min. Extracted with ethyl acetate and water, the organic phase was concentrated under reduced pressure to obtain a pale yellow solid crude product MC126 (1.32 g, 88% yield).

In step d, MC91 (40 mg, 0.12 mmol, 1 eq), crude MC126 (88 mg, 0.36 mmol, 3 eq), and pyridine (28 mg, 0.36 mmol, 3 eq) were dissolved in DCM. The temperature was raised to 50 °C, and the reaction was stirred for 12 h. The solvent was then removed by rotary evaporation under reduced pressure. The resulting concentrate was purified by silica gel column chromatography using DCM:MeOH (20:1-10:1) to obtain a brown oily liquid compound MC127 (25 mg, 37% yield). [M+H] ⁺ 559.2, found: 559.2.

In step e, MC127 (25 mg, 0.045 mmol, 1 eq), NaOH (20 mg, 0.45 mmol, 10 eq) and tetrabutylammonium hydrogen sulfate (8 mg, 0.023 mmol, 0.5 eq) were dissolved in tetrahydrofuran and stirred at room temperature for 3 h. Reduced pressure concentration was performed, and the concentrate was separated and purified by prep-HPLC to obtain a brownish-yellow solid compound 115. ^¹H NMR (400MHz, MeOD- _d₄ ) δ 9.00 (brs, 1H), 8.22 (brs, 1H), 7.96 (brs, 1H), 7.57 (brs, 1H), 7.25 (t, J = 7.9Hz, 1H), 7.19 (d, J = 7.3Hz, 1H), 6.93 (brs, 1H), 3.82 (s, 3H), 3.67 (m, 2H), 3.37 (m, 2H), 3.08 (d, J = 11.4Hz, 2H), 2.99 (s, 1H), 2.64 (m, 2H), 2.03 (m, 2H), 1.96 (m, _{4H ) .} LRMS (ESI) calcd for _{C₂₂H₂₈FN₆O₃} S[M+H] ⁺ 475.2, found:475.3.

Example 116

4-(N-(3-(6-fluoropyridin-3-yl)-2-(5-(4-methyl-4H-1,2,4-triazol-3-yl)-1,2-oxazin-2-yl)phenyl)sulfanyl)butyric acid

4-(N-(3-(6-fluoropyridin-3-yl)-2-(5-(4-methyl-4H-1,2,4-triazol-3-yl)-1,2-oxazinan-2-yl)phenyl)sulfamoyl)butanoic acid

The synthetic route for compound 116 is shown in the figure above. The synthetic method is the same as that for compound 115. The data for compound 116 are as follows: ^¹H NMR (400MHz, MeOD- _d⁴ ) δ 9.02 (brs, 1H), 8.22 (brs, 1H), 7.96 (brs, 1H), 7.58 (brs, 1H), 7.26 (t, J = 7.8Hz, 1H), 7.19 (d, J = 8.6Hz, 1H), 6.93 (brs, 1H), 3.83 (s, 3H), 3.32 (m, 4H), 3.11 (m, 2H), 3.00 (s, 1H), 2.64 (m, 2H), 2.51 (m, 2H), 2.10 (m, 2H), 1.97 (m, _{2H ) .} LRMS (ESI) calcd for _{C₂³H₂₈FN₆O₄} S[M+H] ⁺ 503.2,found:503.2.

Example 117

6-(N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)sulfanyl)hexanoic acid (compound 119)

6-(N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)sulfamoyl)hexanoic acid

The synthetic route for compound 117 is shown in the figure above. The synthetic method is the same as that for compound 115. The data for compound 117 are as follows: ^¹H NMR (600MHz, MeOD- _d4) )δ8.92(brs,1H),8.22(brs,1H),7.97(brs,1H),7.58(brs,1H),7.26(t,J＝7.8Hz,1H),7.19(d,J＝6.4Hz,1H),6.92(brs,1H),3.81(brs,3H),3.3 2(brs,4H),3.07(m,J＝12.5Hz,2H),2.95(s,1H),2.65(m,2H),2.27(m,2H),1.97(m,2H),1.85(m,2H),1.61(m,2H),1.50(m,2H).LRMS(ESI)calcd for C ₂₅ H ₃₁ FN ₆ O ₄ S[M+H] ⁺ 503.2, found: 503.2.

Example 118

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-1H-pyrazole-4-sulfonamide (compound 118)

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-1H-pyrazole-4-sulfonamide

The synthetic route for compound 118 is shown in the figure above. The synthetic method is the same as that for compound 115. The data for compound 118 are as follows: ^¹H NMR (600MHz, MeOD-d4) δ 9.02 (brs, 1H), 8.16 (brs, 1H), 7.95 (m, J = 54.3 Hz, 3H), 7.65 (brs, 1H), 7.24 (brs, 1H), 7.18 (brs, J = 8.3, 2.7 Hz, 1H), 6.93 (brs, 1H), 3.84 (s, 3H), 2.94 (s, 1H), 2.78 (m, 2H), 2.57 (m, 2H), 1.94 (m, 4H). LRMS (ESI) calcd for C ₂₂ H ₂₄ FN ₈ O ₂ S [M+H] ⁺ 483.2, found: 483.2.

Example 119

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)-1-methyl-1H-pyrazole-4-sulfonamide

The synthetic route for compound 119 is shown in the figure above. The crude MC91 obtained above (50 mg, 0.14 mmol, 1.0 eq) was dissolved in DCM (2.0 mL), and 1-methyl-1H-pyrazole-4-sulfonyl chloride (38 mg, 0.21 mmol, 1.5 eq) and Py (17 mg, 0.21 mmol, 1.5 eq) were added at 0 °C. The reaction was heated to 50 °C and stirred for 12 hours. After the reaction was complete, the solvent was removed under reduced pressure, and the product was purified by prep-HPLC to obtain compound 101 as a white solid product (30 mg, yield 43%). ¹ H NMR(400MHz,MeOD-d4)δ8.99(brs,1H),8.16(m,2H),7.91(brs,1H),7.72(brs,1H),7.61(brs,1H),7.22(brs,1H),7.16(brs,J =8.4,2.3Hz,1H),6.91(s,1H),3.91(s,3H),3.82(s,3H),2.92(s,1H),2.79(m,2H),2.55(m,2H),1.92(m,4H).LRMS(ESI)calcd for C ₂₃ H ₂₆ FN ₆ O ₄ S[M+H] ⁺ 497.2,found:497.2.

Example 120

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)thiophene-3-sulfonamide (compound 120)

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

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data of compound 120 are as follows: ^¹H NMR (400MHz, MeOD- _d₄ ) δ 9.17 (s, 1H), 8.13-7.88 (m, 2H), 7.61 (brs, 2H), 7.32-6.92 (m, 4H), 3.86 (s, 3H), 2.98-2.53 (m, 5H), 2.03-1.30 ( _m , 4H) _. LRMS (ESI) calcd for _{C₂₃H₂₄FN₆O₂S₂} [M+H] _{⁺ 499.1} ^, found: 499.2.

Example 121

9-(N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)sulfanyl)-N-hydroxynonanoamide

9-(N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-methyl-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)sulfamoyl)-N-hydroxynonanamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 121 are: LRMS(ESI)calcd for C ₂₈ H ₃₉ FN ₇ O ₄ S[M+H] ⁺ 588.3,found:589.3.

Example 122

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 122 are as follows: ^¹H NMR (600MHz, MeOD) δ 8.97(s, 1H), 8.24(s, 1H), 7.98(s, 1H), 7.59(s, 1H), 7.28(t, J = 7.9Hz, 1H), 7.21(s, 1H), 6.94(s, 1H), 3.84(s, 3H), 3.09(d, J = 11.8Hz, 2H), 2.68(s, 2H), 2.32(s, 1H), 2.12(s, 2H), 1.99(s, 6H), 1.58(d, J = 85.4Hz, 6H). LRMS (ESI) calcd for C ₂₆ H ₃₅ FN ₇ O ₄ S[M+H] ⁺ 546.2, found: 546.3.

Example 123

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 123 are as follows: ^¹H NMR (600MHz, MeOD) δ 9.03 (brs, 1H), 8.22 (brs, 1H), 7.96 (brs, 1H), 7.59 (brs, 1H), 7.26 (t, J = 7.9Hz, 1H), 7.20 (brs, 1H), 6.93 (brs, 1H), 3.85 (s, 3H), 3.32 (m, 4H), 3.08 (m, 3H), 2.64 (m, 2H), 2.29 (m, 2H), 2.13 (m, 2H), 1.99 (m, 2H). LRMS (ESI) calcd for C ₂₃ H ₂₉ FN ₇ O ₄ S [M+H] ⁺ 518.2, found: 518.2.

Example 124

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-(methyl-d3)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methylamino-1-sulfonamide (compound 124)

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-(methyl-d3)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)methylamino-1-sulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 124 are: LRMS(ESI)calcd for C ₂₃ H ₂₇ D ₃ FN ₆ O ₂ S[M+H] ⁺ 476.2, found: 476.3.

Example 125

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-(methyl-d3)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)thiophene-2-sulfonamide (compound 125)

N-(3-(6-fluoropyridin-3-yl)-2-(4-(4-(methyl-d3)-4H-1,2,4-triazol-3-yl)piperidin-1-yl)phenyl)thiophene-2-sulfonamide

The synthetic route is shown in the figure above. The synthetic method is the same as in Example 33. The data for compound 125 are: LRMS(ESI)calcd for C ₂₃ H ₂₁ D ₃ FN ₆ O ₂ S ₂ [M+H] ⁺ 502.1,found:502.2.

Example 1

The method for testing the QPCTL enzyme (also known as gQC or isoQC) inhibitory activity of the compound is as follows: The reaction system consists of QPCTL protease, the fluorescent substrate glutamine 7-amino-4-methylcoumarin (Gln-AMC), the compound, and pyroglutamyl aminopeptidase I (PGPEP I). The reaction is carried out in a black 384-well plate. First, 12.5 μL of QPCTL protease (concentration 5.7 ng/μL) and 2.5 μL of the compound at different concentrations are 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) is added and reacted in a shaker at 37°C and 100 rpm for 20 minutes. Finally, 25 μL of PGPEP I (final concentration 3.97 ng/μL) is added to the system and reacted in a shaker at 37°C and 100 rpm for 30 minutes. The fluorescence intensity of the microplates was read using a TECAN Infinite 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, where A ≤ 10 nM, 10 nM < B ≤ 100 nM, 100 nM < C ≤ 1000 nM, and D > 1000 nM. The activity data are shown in Table 1 below.

Table 1. QPCTL activity data for representative compounds

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. Further research can be conducted on its anti-tumor and other therapeutic effects.

Example 2

The method for testing the QPCT enzyme (also known as sQC) inhibitory activity of the compound is as follows: The reaction system consists of QPCT protease, the fluorescent substrate glutamine 7-amino-4-methylcoumarin (Gln-AMC), the compound, and pyroglutamyl aminopeptidase I (PGPEP I). The reaction is carried out in a black 384-well plate. First, 12.5 μL of QPCT protease (concentration 3.82 ng/μL) and 2.5 μL of the compound at different concentrations are 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) is added, and the reaction is carried out in a shaker at 37°C and 100 rpm for 20 minutes. Finally, 25 μL of PGPEP I (final concentration 3.97 ng/μL) is added to the system, and the reaction is carried out in a shaker at 37°C and 100 rpm for 30 minutes. The fluorescence intensity of the microplates was read using a TECAN Infinite 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, where A ≤ 10 nM, 10 nM < B ≤ 100 nM, and 100 nM < C ≤ 1000 nM. The activity data are shown in Table 2 below.

Table 2. QPCT activity data for representative compounds

Based on the above experimental results, it can be seen that the compound of the present invention has excellent QPCT enzyme inhibitory activity and can be used as a glutamine cyclase inhibitor. Further research will be conducted on its therapeutic effects on Alzheimer's disease and anti-tumor diseases.

Example 3

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

The inhibitory activity of the compounds 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 compounds for 48 hours, the in vitro activity of the compounds 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 compounds, 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 then 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 compounds. The activity data of representative compounds are shown in Table 3 below.

Table 3. Cell activity data representing compounds

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.

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.

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.

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 (15)

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 from 0 to 3, and n is any integer from 0 to 3; X is a carbon atom or a nitrogen atom; _R1 is a 5-10 heteroaryl group optionally substituted by one or more _R7s ; _R2 can be cyano, nitro, -C(=O)N( _R8 ) ₂ , _-NR8C (=O) _R9 , -S(=O) _{2R9 or -NR8S} (=O) _{2R9 ; R3} is a hydrogen atom, a halogen atom, a trifluoromethyl atom, a -NR ₈ C(=O)R ₉ , or a C _1-20 alkoxy atom; _R4 is a 5-10 heteroaryl group optionally substituted with one or more _C1-20 alkyl or amino groups; _R5 is a hydrogen atom, a halogen atom, a hydroxyl group, a _C1-20 alkyl group, or a _C1-20 alkoxy group; optionally, when _R5 is a _C1-20 alkyl group, 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 R ₇ is independently a halogen atom, a _C1-20 alkyl group, a _C1-20 alkyl group optionally substituted with a _{C3-10 cycloalkyl} group, a _C1-20 alkoxy group optionally substituted with a _C3-10 cycloalkyl group, or a _C3-10 cycloalkyloxy group optionally substituted with a _C1-20 alkyl group; Each R ₈ is independently a hydrogen atom or a _C1-20 alkyl group; Each R ₉ is independently a hydrogen atom, a C _1-20 alkyl group, a C _1-20 haloalkyl group, a -NHC _1-20 alkyl group, a -C _1-20 alkylene group, a -C _1-20 alkoxy group, a C _3-10 cycloalkyl group, a 5-10 heterocyclic alkyl group, a 5-10 heteroaryl group, or a C _6-10 aryl group; each of the C _1-10 alkyl group, C _3-10 cycloalkyl group, 5-10 heterocyclic alkyl group, 5-10 heteroaryl group, or C _6-10 aryl group is independently optionally substituted by one or more halogen atoms, C _1-20 alkyl groups or -C(=O)OC _1-20 alkyl groups, carboxyl groups, or hydroxyl groups; Alternatively, _R8 and _R9 can be connected to form a 5-10 element ring structure. The compound according to claim 1, or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug, is characterized in that, m is 0 or 1, n is 0 or 1; X is a carbon atom or a nitrogen atom; _R1 is a 5-10 nitrogen-containing heteroaryl group optionally substituted by one or more _R7 ; _R2 can be cyano, nitro, -C(=O)N( _R8 ) ₂ , _-NR8C (=O) _R9 , -S(=O) _{2R9 or -NR8S} (=O) _{2R9 ; R3} is a hydrogen atom, a halogen atom, a trifluoromethyl atom, _-NR8C (=O) _R9 , or a _C1-10 alkoxy atom. _R4 is a 5-10 nitrogen-containing heteroaryl group optionally substituted with one or more _C1-10 alkyl or amino groups; _R5 is a hydrogen atom, a halogen atom, a hydroxyl group, a _C1-10 alkyl group, or a _C1-10 alkoxy group; optionally, when _R5 is a _C1-10 alkyl group, 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 R ₇ is independently a halogen atom, a _C1-10 alkyl group, a _C1-10 alkyl group optionally substituted with a _C3-6 cycloalkyl group, a _C1-10 alkoxy group optionally substituted with a _C3-6 cycloalkyl group, or a _C3-6 cycloalkyloxy group optionally substituted with a _C1-10 alkyl group. Each R ₈ is independently a hydrogen atom or a _C1-10 alkyl group; Each R ₉ is independently a hydrogen atom, a C _1-10 alkyl group, a C _1-10 haloalkyl group, a -NHC _1-10 alkyl group, a -C _1-10 alkylene group, a -C _1-10 alkoxy group, a C _3-6 cycloalkyl group, a 5-6 membered heterocycloalkyl group, a 5-10 membered heteroaryl group, or a C _6-10 aryl group; each of the C _1-10 alkyl group, C _3-6 cycloalkyl group, 5-6 membered heterocycloalkyl group, 5-10 membered heteroaryl group, or C _6-10 aryl group is independently optionally substituted by one or more halogen atoms, C _1-10 alkyl groups or -C(=O)OC _1-10 alkyl groups, carboxyl groups, or hydroxyl groups; Alternatively, _R8 and _R9 can be connected to form a 5-6 element ring structure. The compound according to claim 2, or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug, is 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; X is a carbon atom or a nitrogen atom; _R1 is a pyridinyl group optionally substituted with one or more _R7 groups; _R2 can be cyano, nitro, -C(=O)N( _R8 ) ₂ , _-NR8C (=O) _R9 , -S(=O) _{2R9 or -NR8S} (=O) _{2R9 ; R3} is a hydrogen atom, a halogen atom, a trifluoromethyl atom, _-NR8C (=O) _R9 , or a methoxy atom; _R4 is optionally substituted with one or more of the following groups, either _C1-10 alkyl or amino groups: in, Indicates the combination of keys; _R5 is a hydrogen atom, a halogen atom, a hydroxyl group, or a _C1-10 alkyl group; optionally, when _R5 is a _C1-10 alkyl group, 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 R ₇ is independently substituted with a methyl, methoxy, or cyclopropyl methoxy or halogen atom; Each R ₈ is independently a hydrogen atom or a _C1-10 alkyl group; Each R ₉ is independently a hydrogen atom, a C _1-10 alkyl group, a C _1-10 haloalkyl group, a -NHC _1-10 alkyl group, a -C _1-10 alkylene group, a -C _1-10 alkoxy group, a C _3-6 cycloalkyl group, a 5-6 membered heterocycloalkyl group, a 5-10 membered heteroaryl group, or a C _6-10 aryl group; each of the C _1-10 alkyl group, C _3-6 cycloalkyl group, 5-6 membered heterocycloalkyl group, 5-10 membered heteroaryl group, or C _6-10 aryl group is independently optionally substituted by one or more halogen atoms, C _1-10 alkyl groups or -C(=O)OC _1-10 alkyl groups, carboxyl groups, or hydroxyl groups; Alternatively, _R8 and _R9 can be connected to form a five-element ring structure. The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof, is characterized in that the compound is as shown in formula (II):
Wherein, m, n, X, _R1 , _R2 , _R3 , _R4 , _R5 and _R6 have the definitions as described in any one of claims 1-3. The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof, is characterized in that the compound is as shown in formula (III):
Wherein, m, n, X, _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. The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof, is characterized in that the compound is as shown in formula (IV):
Wherein, m, n, X, _R1 , _R2 , _R3 , _R4 , _R5 and _R6 have the definitions as described in any one of claims 1-3. The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof, is characterized in that the compound is as shown in formula (V):
Wherein, X, _R1 , _R2 , _R3 , _R4 , _R5 and _R6 have the definitions as described in any one of claims 1-3. The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof, is characterized in that the compound is as shown in formula (VI) or (VII):
Wherein, m, n, X, _R1 , _R4 , _R5 , _R8 and _R9 have the definitions as described in any one of claims 1-3; Preferably, _R4 is a group optionally substituted with one or two _C1-10 alkyl or amino groups, such as the following groups: in, Indicates the combination of keys; Preferably, _R5 is a hydrogen atom or a halogen atom. The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof, is characterized in that the compound is as shown in formula (VIII):
Wherein, m, n, X, _R1 , _R2 , _R3 , _R5 and _R6 have the definitions as described in any one of claims 1-3; Z is a carbon atom or a nitrogen atom; _R10 is a hydrogen atom, a _C1-10 alkyl or _C1-10 alkoxy-substituted benzyl group; optionally, _R5 and _R10 are linked to form a 5-10 membered ring structure; R ₁₁ is a hydrogen atom, a _C1-10 alkyl group, or an amino group. The compound according to claim 9, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof, is characterized in that _R1 is one of the following groups:
in,
Indicates the combination of keys; Each R ₁₂ is an independent halogen atom; R ₁₃ is a methoxy or halogen atom substituted with methyl, methoxy, or cyclopropyl groups; Each R ₁₄ is independently a _C1-10 alkyl group. The following are nitrogen-containing heterocyclic compounds or their pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, isotope labels, or prodrugs:




A pharmaceutical composition comprising, as an active ingredient, a nitrogen-containing heterocyclic compound according to any one of claims 1-11 or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug thereof; preferably, the pharmaceutical composition contains at least one pharmaceutically acceptable carrier. A combination of drugs comprising a nitrogen-containing heterocyclic compound according to any one of claims 1-11 or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label or prodrug or a pharmaceutical composition according to claim 12, and an antibody; preferably, the antibody is a PD-1/PD-L1 antibody or a CD47 antibody. The use of a nitrogen-containing heterocyclic compound or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, isotope label, or prodrug according to any one of claims 1-11, or the pharmaceutical composition according to claim 12, or the combination of drugs according to claim 10, in the inhibition of glutamine cyclase. 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 1-11, or the pharmaceutical composition according to claim 12, or the use of 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, obesity 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.