CN107759587B

CN107759587B - [1,2,4] triazolo [1,5-a ] pyridine compound and preparation method and medical application thereof

Info

Publication number: CN107759587B
Application number: CN201710705571.9A
Authority: CN
Inventors: 殷惠军; 闫旭; 宗利斌; 董流欣; 韩亚超; 郗青川; 窦浩帅; 杨琰
Original assignee: National Institutes of Pharmaceutical R&D Co Ltd
Current assignee: National Institutes of Pharmaceutical R&D Co Ltd
Priority date: 2016-08-19
Filing date: 2017-08-17
Publication date: 2021-01-26
Anticipated expiration: 2037-08-17
Also published as: CN107759587A

Abstract

The present invention relates to [1,2,4]]Triazolo [1,5-a]Pyridine compounds, and a preparation method and medical application thereof. In particular, the invention relates to a compound shown in a general formula I, a preparation method thereof, a pharmaceutical composition containing the compound, an application of the compound as a JAK kinase inhibitor, the compound and a medicament containing the compoundThe compositions may be used to treat diseases associated with JAK kinase activity, such as inflammation, autoimmune disorders, cancer, and the like. Wherein the definition of each substituent in the general formula I is the same as that in the specification.

Description

[1,2,4] triazolo [1,5-a ] pyridine compound and preparation method and medical application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a novel [1,2,4] triazolo [1,5-a ] pyridine compound, a preparation method thereof, a pharmaceutical composition containing the compound, and application of the compound in regulating the activity of Janus kinase (JAK) and treating and/or preventing diseases related to the activity of JAK.

Background

Intracellular signaling processes are an efficient way for cells to respond to external stimuli and ultimately elicit specific biological effects. Cytokines are capable of intracellular signaling through a variety of signal transduction pathways, thereby being involved in the regulation of hematopoietic functions and many important biological functions associated with immunity. The Janus kinase (JAK) family of protein tyrosine kinases and the activator of transcription (STAT) play important roles in cytokine signaling.

The Janus kinase (JAK) family plays a role in cytokine-dependent regulation of cellular proliferation and function involved in the immune response. Currently, there are four known mammalian JAK family members: jak1 (also known as Janus kinase-1), Jak2 (also known as Janus kinase-2), Jak3 (also known as Janus kinase, leukocyte; JAKL 1; L-JAK and Janus kinase-3), Tyk2 (also known as protein-tyrosine kinase 2). Jak1, Jak2, and Tyk2 are widely present in various tissues and cells, while Jak3 is only present in the bone marrow and lymphatic system.

Tyk2 was the first JAK kinase discovered and plays an important role in regulating the biological response of IL-12 and bacterial Lipopolysaccharide (LPS), and is also involved in IL-6, IL-10 and IL-12 mediated signal transduction pathways. Targeting Tyk2 could be a new strategy for treating IL-12-, IL-23-, or type I IFN-mediated diseases, including but not limited to rheumatoid arthritis, multiple sclerosis, lupus, psoriasis, psoriatic arthritis, inflammatory bowel disease, uveitis, sarcoidosis, and cancer.

Jak1 plays an important role in regulating the biological response functions of a variety of cytokine receptor families. Jak1 knockout mice have an early postnatal lethal factor phenotype and the nervous system is also compromised, resulting in congenital defects in young mice. The study shows that the mice with the Jak1 gene knockout can generate secretion defects of thymocytes and B cells, and the tissue with the Jak1 gene knockout has obviously weakened response to LIF, IL-6 and IL-10. Clinical trials have shown that selective Jak1 inhibitors also have RA-ameliorating effects in clinical studies, and the Jak1 inhibitor ABT-494 in phase III has positive results in two trials involving rheumatoid arthritis patients who do not respond adequately to methotrexate or one of the Tumor Necrosis Factor (TNF) blockers.

Jak2 plays an important role in Epo, IL-3, GM-CSF, IL-5, Tpo and IFN γ -mediated signal transduction. Jak2 knockout mice have an embryonic lethal factor phenotype and die at day 12.5 of pregnancy due to defective erythropoiesis. Similar phenomena were also observed in Epo knock-out mice, indicating that Epo is closely related to Jak2 kinase activity. Jak2 kinase is not involved in IL-23 and IL-14 receptor family mediated signal transduction. Studies have shown that Jak2 kinase does not respond to IFN γ, but responds to IFN α and IL-6. The mutant Jak2 protein is capable of activating downstream signals in the absence of cytokine stimulation, resulting in spontaneous growth and/or hypersensitivity to cytokines which are believed to play a contributing role in the processes of these diseases. Inhibitors of Jak2 have been described as having therapeutic effects on proliferative diseases.

Jak3 plays an important role in a variety of biological processes, such as lymphocyte proliferation processes, IgExtent receptor mediated mast cell degeneration, prevention of T cell activation, and involvement in signal transduction mediated by all γ C families, including IL-23, IL-4, IL-7, IL-9, IL-15, and IL-21. Jak3 kinase function is not the same in humans and mice, e.g., Severe Combined Immunodeficiency Disease (SCID) patients have normal B cells but lack T cell function. This is because IL-7 plays an important role in B cell proliferation in mice but does not affect B cell proliferation in humans. The Jak3 gene knocks out SCID phenotype of mammals and the specific expression of JAK lymphocytes, making Jak3 an immunosuppressant target. Based on the role of Jak3 in regulating lymphocytes, targeting Jak3 and Jak3 mediated pathways can be useful in the treatment of autoimmune diseases.

After the cytokine binds to the receptor, the receptor forms a dimer, and JAKs coupled to the receptor approach each other and are activated by phosphorylation of tyrosine residues. And then catalyzes phosphorylation of tyrosine residues of the receptor itself to form a "docking site". Signal Transducers and Activators of Transcription (STATs) are a group of cytoplasmic proteins that regulate binding of DNA to target genes. STAT families include STAT1, STAT2, STAT3, STAT4, STAT5a, STAT5b, and STAT 6. STAT recognizes the "docking site" through SH2 domain and is activated by phosphorylation of its C-terminal tyrosine residue by JAK kinases. The activated Stat factor is transferred into the nucleus and plays an important role in regulating innate and adaptive host immune responses.

Activation of the JAK/STAT signaling pathway contributes to the development of a variety of diseases, including, but not limited to, many aberrant immune responses, such as allergy, asthma, rheumatoid arthritis, amyotrophic lateral sclerosis, and multiple sclerosis. It is also associated with cancers such as leukemia (acute myeloid leukemia and acute lymphocytic leukemia), solid tumors (uterine leiomyosarcoma, prostate cancer), and the like.

Rheumatoid Arthritis (RA) is an autoimmune disease characterized by inflammation and destruction of joint structures. When the disease is not treated effectively, substantial disability and pain, and even premature death, result from loss of joint functionality. The aim of RA treatment is therefore not only to delay the progression of the disease but also to obtain a reduction in symptoms, thereby terminating joint destruction. The global prevalence of RA is about 0.8%, with women having a three-fold prevalence rate over men. RA is difficult to treat, there is currently no cure, and treatment focuses on relieving pain and preventing diseased joint degeneration. Clinical treatment strategies include nonsteroidal anti-inflammatory drugs (NSAIDs), hormones, disease-modifying antirheumatic drugs (DMARDS), and biologic drugs, mainly to relieve the symptoms of joint damage and swelling. Clinical application of DMARDS (such as methotrexate, hydroxychloroquine, leflunomide, sulfasalazine) and DMARDS has better effect when being combined with biological drugs. Despite the abundance of anti-RA drugs, pain still exists in more than 30% of patients. Recent studies have shown that intervention of the JAK/STAT signaling pathway is a new approach to RA treatment.

Tofacitinib is the first novel FDA-approved oral JAK inhibitor that acts on JAK1 and JAK3, and is a small molecule compound useful in the treatment of RA. Clinical trials indicate that tofacitinib exhibits a therapeutic effect that is not inferior to TNF inhibitors. The combined use of Methotrexate (MTX) and tofacitinib also has certain therapeutic effects on patients who are not responsive to TNF inhibitors. Therefore, tofacitinib is recommended for clinical first-line single drug administration, and has a therapeutic advantage compared with MTX. Increased phosphorylation of STAT1 and STAT3 was found in the synovial fluid of tofacitinib-treated patients, suggesting that it is primarily through intervention in the JAK/STAT signaling pathway. However, tofacitinib is in relief of RAThe symptoms can also bring about some side effects, and certain infections, malignant tumors and lymphomas are caused. Serious infections and malignancy-induced adverse reactions also have been reported during biopharmaceutical treatment of RA, and novel safety data suggest that the overall risk of infection and mortality for tofacitinib is similar to that of biological agents for treating RA. Given the pleiotropic nature of JAKs in many regulatory pathways and immune processes, non-selective JAK inhibitors carry risks of adverse effects, such as hypercholesterolemia and infection. Selective JAK inhibitors are an important direction of current research. Filgotinib, from Galapagos, Belgium, is a new generation of JAK1 selective inhibitor with reduced risk of anemia or infection with tofacitinib. In a recently completed clinical phase II trial on moderate to severe RA patients who do not respond adequately to methotrexate treatment, the primary endpoint reached after 12 weeks of Filgotinib treatment-80% for ACR20, with a 200mg dose showing statistical significance; at all dose levels ACR50 response and DAS28 reduction were statistically significant compared to controls; the safety level was similar to before, with good tolerability. After 24 weeks, 64% of patients achieved DAS28 remission or low activity; all doses of ACR50 response, ACR70 response and DAS28 reduction showed statistically significant levels, reaching 39% ACR 70. However, Filgotinib was relatively weak against JAK1 IC₅₀Above 10nM, the clinical dose is also relatively high.

RA is a very heterogeneous disease and the therapeutic application of suitable drugs to RA patients is a major challenge. Although a range of JAK inhibitors have been disclosed, there is still a need to develop compounds with better selectivity and potency. Thus, there is a continuing need for new or improved agents that inhibit kinases such as Janus kinases for the development of new, more potent drugs for the treatment of RA or other JAK-associated diseases.

Disclosure of Invention

The inventor designs and synthesizes a series of compounds containing [1,2,4] triazolo [1,5-a ] pyridine skeleton through intensive research, screens the JAK kinase activity, and shows that the compounds have outstanding anti-JAK kinase activity and can be developed into medicines for treating diseases related to the JAK kinase activity.

Therefore, the invention aims to provide a compound shown in the general formula I or a racemate, an enantiomer, a diastereoisomer, a mixture form, a prodrug or a pharmaceutically acceptable salt thereof,

wherein:

R¹selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -C (O) R^a、-C(S)R^a、-C(O)OR^a、-S(O)R^a、-S(O)₂R^aWherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally further substituted with one or more R²Substitution;

each R²Each independently selected from the group consisting of halogen, amino, nitro, cyano, hydroxy, mercapto, oxo, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, -C (O) R^a、-O(O)CR^a、-C(O)OR^a、-C(O)NR^aR^b、-NR^aR^b、-NHC(O)R^a、-S(O)R^a、-S(O)₂R^a、-S(O)NR^aR^b、-S(O)₂NR^aR^b、-NHS(O)R^a、-NHS(O)₂R^a(ii) a Wherein said alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxy, mercapto, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

cy is selected from aryl and heteroaryl;

each R³Independently selected from hydrogen, halogen, alkyl, alkoxy, haloalkyl, haloalkoxy;

R^4aand R^4bEach independently selected from hydrogen, halogen, amino, nitro, cyano, hydroxy, mercapto, oxo, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein said alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxy, mercapto, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

R⁵selected from cycloalkyl, heterocyclyl, aryl, heteroaryl; said cycloalkyl, heterocyclyl, aryl or heteroaryl being optionally further substituted by one or more R⁶Substitution;

each R⁶Each independently selected from the group consisting of halogen, amino, nitro, cyano, hydroxy, mercapto, oxo, imino, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -C (O) R^a、-O(O)CR^a、-C(O)OR^a、-C(O)NR^aR^b、-NR^aR^b、-NHC(O)R^a、-S(O)R^a、-S(O)₂R^a、-S(O)NR^aR^b、-S(O)₂NR^aR^b、-NHS(O)R^a、-NHS(O)₂R^a(ii) a Wherein said alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxy, mercapto, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

R^aand R^bEach independently selected from hydrogen, halogen, hydroxy, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, hydroxy, mercapto, carboxyl, ester, oxo, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

n1 is an integer from 1 to 4;

m1 is an integer of 1 to 4.

In a preferred embodiment of the present invention, the compounds of formula I according to the present invention or their racemates, enantiomers, diastereomers, or mixtures thereof, prodrugs thereof or pharmaceutically acceptable salts thereof, wherein Cy is selected from C₆～C₁₀Aryl or 6-to 10-membered heteroaryl, preferably phenyl or pyridyl.

In another preferred embodiment of the present invention, the compound of formula I according to the present invention, or its racemate, enantiomer, diastereomer, or mixture thereof, or its prodrug, or its pharmaceutically acceptable salt, is a compound of formula II, or its racemate, enantiomer, diastereomer, or mixture thereof, or its prodrug, or its pharmaceutically acceptable salt, as follows:

wherein R is¹、R^4a、R^4bAnd R⁵As defined in formula I.

In another preferred embodiment of the present invention, the compounds of formula I according to the present invention or their racemates, enantiomers, diastereomers, or their mixtures, prodrugs or pharmaceutically acceptable salts thereof, wherein R is^4bAnd R^4cEach independently selected from hydrogen, halogen, alkyl, said alkyl being optionally further substituted by one or more groups selected from halogen, amino, nitro, cyano, hydroxy, mercapto, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl. In another preferred embodiment of the present invention, the compound of formula I according to the present invention, which is a compound of formula III or its racemate, enantiomer, diastereomer, or mixture thereof, prodrug thereof, or pharmaceutically acceptable salt thereof, is represented by formula IA racemate, enantiomers, diastereomers, or mixtures thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof:

wherein R is¹And R⁵As defined in formula I.

In another preferred embodiment of the present invention, the compound of formula I according to the present invention, R, or a racemate, enantiomer, diastereomer or a mixture thereof, a prodrug thereof or a pharmaceutically acceptable salt thereof, R⁵Selected from cycloalkyl, heterocyclyl, preferably C₃～C₆Cycloalkyl or 4-to 7-membered heterocycloalkyl; said cycloalkyl, heterocyclyl is optionally further substituted by one or more R⁶Substitution;

wherein each R is⁶Each independently selected from the group consisting of halogen, amino, nitro, cyano, hydroxy, mercapto, oxo, imino, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -C (O) R^a、-O(O)CR^a、-C(O)OR^a、-C(O)NR^aR^b、-NR^aR^b、-NHC(O)R^a、-S(O)R^a、-S(O)₂R^a、-S(O)NR^aR^b、-S(O)₂NR^aR^b、-NHS(O)R^a、-NHS(O)₂R^a(ii) a Wherein said alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally further substituted with one or more groups selected from halogen, amino, nitro, cyano, oxo, hydroxy, mercapto, carboxyl, ester, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl;

R⁶preferably halogen, oxo, alkyl, -C (O) R^a、-S(O)R^a、-S(O)₂R^aA haloalkyl group; wherein R is^aSelected from alkyl groups.

In another preferred embodiment of the invention, according to the inventionA compound shown as a general formula I or a racemate, an enantiomer, a diastereoisomer, a mixture form, a prodrug or a pharmaceutically acceptable salt thereof, wherein R is⁵Selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, thiomorpholinyl, morpholinyl, piperidinyl, piperazinyl, pyrrolidinyl, azaspirocycloalkyl, azacycloheteroalkyl, preferably R⁵The nitrogen atom of the group is attached to the parent nucleus.

In another preferred embodiment of the present invention, the compounds of formula I according to the present invention or their racemates, enantiomers, diastereomers, or their mixtures, prodrugs or pharmaceutically acceptable salts thereof, wherein R is¹Selected from cycloalkyl, heterocyclyl, aryl, heteroaryl, preferably C₃～C₆Cycloalkyl, 4-to 7-membered heterocyclic group, C₆～C₁₀Aryl, 6-to 10-membered heteroaryl, more preferably cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, piperidinyl, piperazinyl, pyrrolidinyl, morpholinyl, thiomorpholinyl, phenyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, dihydrooxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, naphthyl, quinolinyl, isoquinolinyl, quinazolinyl, quinoxalinyl, indazolyl, indolyl, isoindolyl, benzothiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, benzothienyl, benzofuranyl, benzotriazolyl, pyrazolopyridinyl, triazolopyridinyl, imidazopyridinyl, pyrazolopiperazinyl, pyrazolopyridinyl; wherein said cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally further substituted with one or more R²Substitution;

each R²Each independently selected from the group consisting of halogen, amino, oxo, alkyl, alkoxy, cycloalkyl, heterocyclyl, -C (O) R^a、-S(O)R^a、-S(O)₂R^a、-S(O)NR^aR^b、-S(O)₂NR^aR^b(ii) a Wherein said alkyl, alkoxy, cycloalkyl, heterocyclyl or heteroaryl isOptionally further substituted by one or more groups selected from halogen, hydroxy, alkyl, alkoxy;

R^aand R^bEach independently selected from hydrogen, hydroxy, alkyl, cycloalkyl.

In another preferred embodiment of the present invention, the compound of formula I according to the present invention, R, or a racemate, enantiomer, diastereomer or a mixture thereof, a prodrug thereof or a pharmaceutically acceptable salt thereof, R¹Selected from the group consisting of-C (O) R^a、-C(S)R^aor-C (O) NR^aR^b，

R^aAnd R^bEach independently selected from cycloalkyl, heterocyclyl, aryl, heteroaryl, preferably C₃～C₆Cycloalkyl, 4-to 7-membered heterocyclic group, C₆～C₁₀Aryl, 6-to 10-membered heteroaryl, more preferably cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, phenyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, thienyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl; wherein said cycloalkyl, heterocyclyl, aryl, heteroaryl are optionally further substituted by one or more groups selected from halogen, oxo, alkyl, alkoxy, cycloalkyl, heterocyclyl.

Typical compounds of the invention include, but are not limited to:

n- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) -2-oxopyrrolidine-3-carboxamide;

n- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) pyrrolidine-2-carboxamide;

n- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) -1-methylazetidine-3-carboxamide;

n- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) -1H-pyrazole-5-carboxamide;

n- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) thiophene-2-carboxamide;

n- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) -1H-imidazole-2-carboxamide;

n- (5- (4- ((2, 2-dioxo-2-thia-6-azaspiro [3.3] heptan-6-yl) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide;

n- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarbothioamide;

4- (4- (2- ((1-methyl-1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- (oxazol-2-ylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- (thiazol-2-ylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- (cyclopropylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- ((1, 4-dimethyl-1H-pyrazol-5-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- (pyridin-4-ylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- ((1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- ((1-isopropyl-1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

1- (4- ((5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) amino) -1H-pyrazol-1-yl) ethan-1-one;

4- (4- (2- ((2-fluoro-4, 5-dimethoxyphenyl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- ((2-fluoro-4-methoxyphenyl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

3- ((5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) amino) benzenesulfonamide;

4- (4- (2- ((1-cyclopropyl-1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- ((1-methyl-1H-indazol-6-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- ((5-methyl-4, 5,6, 7-tetrahydropyrazolo [1,5-a ] pyrazin-2-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- ((1- (1-methylazetidin-3-yl) -1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- ((1- (1- (ethylsulfonyl) -1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- (cyclopentylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- (isoxazol-4-ylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- (isothiazol-4-ylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- ((1-methylpiperidin-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- (cyclohexylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- ((1- (2,2, 2-trifluoroethyl) -1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- ((1- (difluoromethyl) -1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- ((1- (2-methoxyethyl) -1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

4- (4- (2- ((1- (2-hydroxyethyl) -1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

n- (1-methyl-1H-pyrazol-4-yl) -5- (4- (morpholinomethyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine;

5- (4- ((4, 4-difluoropiperidin-1-yl) methyl) phenyl) -N- (1-methyl-1H-pyrazol-4-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine;

n- (1-methyl-1H-pyrazol-4-yl) -5- (4- (piperidin-1-ylmethyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine;

n- (1-methyl-1H-pyrazol-4-yl) -5- (4- (pyrrolidin-1-ylmethyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine;

n- (1-methyl-1H-pyrazol-4-yl) -5- (4- ((6-methyl-2, 6-diazaspiro [3.3] hept-2-yl) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine;

n- (1-methyl-1H-pyrazol-4-yl) -5- (4- ((5-methyl-hexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine;

n- (1-methyl-1H-pyrazol-4-yl) -5- (4- ((3-methyl-3, 8-diazabicyclo [3.2.1] octan-8-yl) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine;

4- (4- (2- ((4, 4-dimethyl-4, 5-dihydrooxazol-2-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide;

1- (4- (4- (2- ((1-methyl-1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) piperazin-1-yl) ethan-1-one;

n- (1-methyl-1H-pyrazol-4-yl) -5- (4- ((4- (methylsulfonyl) piperazin-1-yl) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine;

or a racemate, enantiomer, diastereomer or mixture thereof, a prodrug thereof or a pharmaceutically acceptable salt thereof.

The present invention also provides a method for preparing a compound represented by the general formula I, or a racemate, enantiomer, diastereomer or mixture thereof, a prodrug thereof or a pharmaceutically acceptable salt thereof, the method comprising:

1) when R is¹is-C (O) R^a、-C(O)OR^a、-S(O)R^a、-S(O)₂R^aWhen the temperature of the water is higher than the set temperature,

reacting the compound Ia with corresponding acyl chloride at low temperature or room temperature under the alkaline condition to obtain a compound of a general formula I, wherein triethylamine is preferably used as an alkaline reagent;

2) when R is¹When the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic radical, aryl and heteroaryl are used,

under the conditions of high-temperature alkaline microwave or high-temperature alkaline tube sealing and under the action of a catalyst, reacting a compound Ib with a corresponding amine compound to obtain a compound shown in a general formula I, wherein an alkaline reagent is preferably sodium tert-butoxide or cesium carbonate, and a catalyst is preferably Pd₂(dba)₃Xantphos, temperature preferably 160 ℃;

3) when R is¹Is C (S) R^aWhen the temperature of the water is higher than the set temperature,

reacting the compound Ic with a Lawson reagent at a high temperature, wherein the temperature is preferably 120 ℃, so as to obtain a compound in the general formula I;

wherein R is¹、R³、R^a、R^4a、R^4b、R⁵Cy, m1, n1 are as defined in claim 1.

Another aspect of the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I or its racemate, enantiomer, diastereomer, or mixture thereof, prodrug thereof, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, or excipient.

The invention also relates to a method for preparing the composition, which comprises the step of mixing the compound shown in the general formula I or the raceme, the enantiomer, the diastereoisomer, the mixture form, the prodrug or the pharmaceutically acceptable salt thereof with a pharmaceutically acceptable carrier, diluent or excipient.

The invention further relates to a compound shown in the general formula I, or a racemate, an enantiomer, a diastereoisomer, a mixture form, a prodrug or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the compound and application thereof in preparing Janus kinase inhibitors.

The invention further relates to a compound shown in the general formula I or a racemate, an enantiomer, a diastereoisomer, a mixture form, a prodrug or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the compound, and application of the compound in preparation of medicines for preventing and/or treating JAK activity-related diseases. The disease associated with JAK activity may be selected from inflammation, autoimmune diseases, or cancer, wherein the inflammation is for example arthritis, in particular rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, uveitis; such autoimmune diseases as multiple sclerosis, lupus, psoriasis, sarcoidosis; such as breast cancer, cervical cancer, colon cancer, lung cancer, stomach cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor, ovarian tumor, peritoneal tumor, melanoma, solid tumor, glioma, glioblastoma, hepatocellular carcinoma, mastoid renal tumor, head and neck tumor, leukemia, lymphoma, myeloma, and non-small cell lung cancer.

The invention also relates to a compound shown in the general formula I or a racemate, an enantiomer, a diastereoisomer, a mixture form, a prodrug or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the same, and application thereof as a Janus kinase inhibitor.

The invention further relates to a compound shown in the general formula I or a racemate, an enantiomer, a diastereoisomer, a mixture form, a prodrug or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the compound, and application of the compound as a medicine for preventing and/or treating diseases related to JAK activity. The disease associated with JAK activity may be selected from inflammation, autoimmune diseases, or cancer, wherein the inflammation is for example arthritis, in particular rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, uveitis; such autoimmune diseases as multiple sclerosis, lupus, psoriasis, sarcoidosis; such as breast cancer, cervical cancer, colon cancer, lung cancer, stomach cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor, ovarian tumor, peritoneal tumor, melanoma, solid tumor, glioma, glioblastoma, hepatocellular carcinoma, mastoid renal tumor, head and neck tumor, leukemia, lymphoma, myeloma, and non-small cell lung cancer.

The present invention further relates to a method of inhibiting Janus kinase comprising administering to a patient in need thereof a therapeutically effective dose of a compound of formula I or its racemates, enantiomers, diastereomers, or mixture thereof, prodrug thereof or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the same.

The present invention further relates to a method for the prevention and/or treatment of a disease associated with JAK activity comprising administering to a patient in need thereof a therapeutically effective dose of a compound of formula I or its racemate, enantiomer, diastereomer or mixture thereof, prodrug thereof or pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the same. The disease associated with JAK activity may be selected from inflammation, autoimmune diseases, or cancer, wherein the inflammation is for example arthritis, in particular rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, uveitis; such autoimmune diseases as multiple sclerosis, lupus, psoriasis, sarcoidosis; such as breast cancer, cervical cancer, colon cancer, lung cancer, stomach cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor, ovarian tumor, peritoneal tumor, melanoma, solid tumor, glioma, glioblastoma, hepatocellular carcinoma, mastoid renal tumor, head and neck tumor, leukemia, lymphoma, myeloma, and non-small cell lung cancer.

The compounds of formula I of the present invention may be used to form pharmaceutically acceptable acid addition salts with acids according to conventional methods in the art to which the present invention pertains. The acid includes inorganic acids and organic acids, and particularly preferably hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid, and the like.

In addition, the invention also comprises a prodrug of the compound shown in the general formula I. Prodrugs of the compounds of formula I are derivatives of the compounds of formula I which may themselves be less active or even inactive, but which, upon administration, are converted to the corresponding biologically active form under physiological conditions (e.g., by metabolism, solvolysis, or otherwise).

The pharmaceutical compositions containing the active ingredient may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Oral compositions may be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions may contain one or more ingredients selected from the group consisting of: sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide a pleasant to the eye and palatable pharmaceutical preparation. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be inert excipients, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as microcrystalline cellulose, croscarmellose sodium, corn starch or alginic acid; binding agents, for example starch, gelatin, polyvinylpyrrolidone or acacia; and lubricating agents, such as magnesium stearate, stearic acid or talc. These tablets may be uncoated or they may be coated by known techniques which mask the taste of the drug or delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, water soluble taste masking substances such as hydroxypropylmethyl cellulose or hydroxypropyl cellulose, or time extending substances such as ethyl cellulose, cellulose acetate butyrate may be used.

Oral formulations may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with a water soluble carrier, for example polyethylene glycol, or an oil vehicle, for example peanut oil, liquid paraffin or olive oil.

Aqueous suspensions contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone and acacia; dispersing or wetting agents may be a naturally occurring phosphatide, for example lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol (heptadecaethyleneoxy cetanol), or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyethylene oxide sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene oxide sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example ethyl or n-propyl paraben, one or more colouring agents, one or more flavouring agents and one or more sweetening agents, such as sucrose, saccharin or aspartame.

Oil suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oil suspension may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable preparation. These compositions can be preserved by the addition of antioxidants such as butylated hydroxyanisole or alpha-tocopherol.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent or one or more preservatives. Suitable dispersing or wetting agents and suspending agents are illustrative of the examples given above. Other excipients, for example sweetening, flavoring and coloring agents, may also be present. These compositions are preserved by the addition of an antioxidant such as ascorbic acid.

The pharmaceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin or mixtures thereof. Suitable emulsifying agents may be naturally-occurring phosphatides, for example soy bean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyethylene oxide sorbitol monooleate. The emulsions may also contain sweetening agents, flavouring agents, preservatives and antioxidants. Syrups and elixirs may be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative, a colorant and an antioxidant.

The pharmaceutical compositions may be in the form of a sterile injectable aqueous solution. Among the acceptable vehicles and solvents that may be employed are water, ringer's solution and isotonic sodium chloride solution. The sterile injectable preparation may be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in the oil phase. For example, the active ingredient is dissolved in a mixture of soybean oil and lecithin. The oil solution is then treated to form a microemulsion by adding to a mixture of water and glycerol. The injection solution or microemulsion may be injected into the bloodstream of a patient by local bulk injection. Alternatively, it may be desirable to administer the solutions and microemulsions in a manner that maintains a constant circulating concentration of the compounds of the present invention. To maintain such a constant concentration, a continuous intravenous delivery device may be used. An example of such a device is an intravenous pump model Deltec CADD-PLUS. TM.5400.

The pharmaceutical compositions may be in the form of sterile injectable aqueous or oleaginous suspensions for intramuscular and subcutaneous administration. The suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension prepared in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any blend fixed oil may be used, including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The compounds of the present invention may be administered in the form of suppositories for rectal administration. These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid in the rectum and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, glycerogelatin, hydrogenated vegetable oils, polyethylene glycols of various molecular weights and mixtures of fatty acid esters of polyethylene glycols.

As is well known to those skilled in the art, the dosage of a drug to be administered depends on a variety of factors, including but not limited to: the activity of the particular compound employed, the age of the patient, the weight of the patient, the health of the patient, the patient's integuments, the patient's diet, the time of administration, the mode of administration, the rate of excretion, the combination of drugs, and the like; in addition, the optimal treatment regimen, such as the mode of treatment, the daily amount of compound (I) of the formula or the type of pharmaceutically acceptable salt, can be verified according to conventional treatment protocols.

The compound containing the [1,2,4] triazolo [1,5-a ] pyridine skeleton and pharmaceutically acceptable salts, hydrates or solvates thereof which are shown in the general formula I can be used as active ingredients, mixed with pharmaceutically acceptable carriers or excipients to prepare a composition, and prepared into clinically acceptable dosage forms. The derivatives of the present invention may be used in combination with other active ingredients as long as they do not produce other adverse effects such as allergic reactions and the like. The compounds of the present invention may be used as the sole active ingredient, or may be used in combination with other drugs for the treatment of diseases associated with JAK activity. Combination therapy is achieved by administering the individual therapeutic components simultaneously, separately or sequentially.

According to the invention, through Jak1, Jak2, Jak3 and Tyk2 activity tests, the compounds have remarkable Janus kinase regulating activity, so that the compounds can be used for treating and/or preventing diseases related to the JAK activity, such as inflammation, autoimmune diseases, cancer or other diseases. In particular for the preparation of a medicament for the treatment and/or prophylaxis of rheumatoid arthritis, psoriasis and/or diseases involving cartilage and bone joint degeneration.

Detailed description of the invention

Unless stated to the contrary, terms used in the specification and claims have the following meanings.

The term "alkyl" refers to a saturated aliphatic hydrocarbon group which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-dimethylpentyl, 2, 4-dimethylpentyl, 2-dimethylpentyl, 3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2, 3-dimethylhexyl, 2, 4-dimethylhexyl, 2, 5-dimethylhexyl, 2-dimethylhexyl, 3-dimethylhexyl, 4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2-dimethylpentyl, 2-dimethylhexyl, 3-dimethylpentyl, 2-ethylhexyl, 3-dimethylhexyl, 2, 2-diethylpentyl, n-decyl, 3-diethylhexyl, 2-diethylhexyl, and various branched isomers thereof. More preferred are lower alkyl groups having 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl and the like. The alkyl group may be substituted or unsubstituted, and when substituted, the substituent may be substituted at any available point of attachment, preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halo, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate.

The term "alkylene" means that one hydrogen atom of an alkyl group is further substituted, for example: "methylene" means-CH₂-, "ethylene" means- (CH)₂)₂-, "propylene" means- (CH)₂)₃-, "butylene" means- (CH)₂)₄-and the like.

The term "alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, e.g., ethenyl, 1-propenyl, 2-propenyl, 1-, 2-or 3-butenyl, and the like. The alkenyl group may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio.

The term "alkynyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond, e.g., ethynyl, propynyl, butynyl, and the like. Alkynyl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio.

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, the cycloalkyl ring containing from 3 to 20 carbon atoms, preferably from 3 to 12 carbon atoms, more preferably from 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyl groups include spiro, fused and bridged cycloalkyl groups.

The term "spirocycloalkyl" refers to a 5 to 20 membered polycyclic group sharing one carbon atom (referred to as a spiro atom) between monocyclic rings, which may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system. Preferably 6 to 14, more preferably 7 to 10. Spirocycloalkyl groups are classified into a single spirocycloalkyl group, a double spirocycloalkyl group or a multi spirocycloalkyl group, preferably a single spirocycloalkyl group and a double spirocycloalkyl group, according to the number of spiro atoms shared between rings. More preferably 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered. Non-limiting examples of spirocycloalkyl groups include:

the term "fused cyclic alkyl" refers to a 5 to 20 membered all carbon polycyclic group in which each ring in the system shares an adjacent pair of carbon atoms with other rings in the system, wherein one or more of the rings may contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system. Preferably 6 to 14, more preferably 7 to 10. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic fused ring alkyls according to the number of constituent rings, preferably bicyclic or tricyclic, more preferably 5-or 6-membered bicycloalkyl. Non-limiting examples of fused ring alkyl groups include:

the term "bridged cycloalkyl" refers to a 5 to 20 membered all carbon polycyclic group in which any two rings share two carbon atoms not directly attached, which may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system. Preferably 6 to 14, more preferably 7 to 10. They may be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl groups, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic, depending on the number of constituent rings. Non-limiting examples of bridged cycloalkyl groups include:

the cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, where the ring to which the parent structure is attached is cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, and the like. Cycloalkyl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate.

The term "heterocyclyl" refers to a saturated or partially unsaturated mono-or polycyclic cyclic hydrocarbon substituent containing from 3 to 20 ring atoms wherein one or more of the ring atoms is selected from nitrogen, oxygen, or S (O)_m(wherein m is an integer of 0 to 2) a hetero atom, excluding a cyclic portion of-O-O-, -O-S-or-S-S-And the remaining ring atoms are carbon. Preferably 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; most preferably 3 to 8 ring atoms, of which 1 to 3 are heteroatoms; most preferably 5 to 6 ring atoms, of which 1-2 or 1-3 are heteroatoms. Non-limiting examples of monocyclic heterocyclyl groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, and the like, preferably 1,2, 5-oxadiazolyl, pyranyl, or morpholinyl. Polycyclic heterocyclic groups include spiro, fused and bridged heterocyclic groups.

The term "spiroheterocyclyl" refers to a 5-to 20-membered polycyclic heterocyclic group in which one atom (referred to as the spiro atom) is shared between monocyclic rings, and in which one or more ring atoms is selected from nitrogen, oxygen, or S (O)_m(wherein m is an integer of 0 to 2) and the remaining ring atoms are carbon. It may contain one or more double bonds, but no ring has a completely conjugated pi-electron system. Preferably 6 to 14, more preferably 7 to 10. The spiro heterocyclic group is classified into a mono-spiro heterocyclic group, a di-spiro heterocyclic group or a multi-spiro heterocyclic group, preferably a mono-spiro heterocyclic group and a di-spiro heterocyclic group, according to the number of spiro atoms shared between rings. More preferred are 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered mono spiroheterocyclic groups. Non-limiting examples of spiro heterocyclic groups include:

the term "fused heterocyclyl" refers to a 5 to 20 membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with other rings in the system, one or more rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system in which one or more ring atoms is selected from nitrogen, oxygen or S (O)_m(wherein m is an integer of 0 to 2) and the remaining ring atoms are carbon. Preferably 6 to 14, more preferably 7 to 10. They may be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups, depending on the number of constituent rings, preferablyIs bicyclic or tricyclic, more preferably a 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic group. Non-limiting examples of fused heterocyclic groups include:

the term "bridged heterocyclyl" refers to a 5 to 14 membered polycyclic heterocyclic group in which any two rings share two atoms not directly attached which may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system in which one or more of the ring atoms is selected from nitrogen, oxygen or S (O)_m(wherein m is an integer of 0 to 2) and the remaining ring atoms are carbon. Preferably 6 to 14, more preferably 7 to 10. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups according to the number of constituent rings, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclic groups include:

the heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring to which the parent structure is attached is heterocyclyl, non-limiting examples of which include:

the heterocyclyl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxy or carboxylate.

The term "aryl" refers to a 6 to 14 membered all carbon monocyclic or fused polycyclic (i.e., rings which share adjacent pairs of carbon atoms) group having a conjugated pi-electron system, preferably 6 to 10 membered, such as phenyl and naphthyl, more preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is an aryl ring, non-limiting examples of which include:

the aryl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxy or carboxylate.

The term "heteroaryl" refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl is preferably 5 to 10 membered, containing 1 to 3 heteroatoms; more preferably 5 or 6 membered, containing 1 to 2 heteroatoms; preferably, for example, imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl and the like, preferably imidazolyl, thiazolyl, pyrazolyl or pyrimidinyl, thiazolyl; more preferably pyrazolyl or thiazolyl. The heteroaryl ring may be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring joined together with the parent structure is a heteroaryl ring, non-limiting examples of which include:

heteroaryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate groups.

The term "alkoxy" refers to-O- (alkyl) and-O- (unsubstituted cycloalkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. Alkoxy groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxy or carboxylate groups.

The term "haloalkyl" refers to an alkyl group substituted with one or more halogens wherein alkyl is as defined above.

The term "haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein the alkoxy group is as defined above.

The term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.

The term "hydroxy" refers to an-OH group.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "amino" refers to the group-NH₂。

The term "cyano" refers to — CN.

The term "nitro" means-NO₂。

The term "oxo" refers to ═ O.

The term "carboxy" refers to-C (O) OH.

The term "mercapto" refers to-SH.

The term "carboxylate" refers to-C (O) O (alkyl) or-C (O) O (cycloalkyl), wherein alkyl is as defined above.

The term "acyl" refers to compounds containing the group-C (O) R, where R is alkyl, cycloalkyl, aryl, heteroaryl.

The term "sulfonic acid group" means-S (O)₂OH。

The term "sulfonate group" means-S (O)₂O (alkyl) or-S (O)₂O (cycloalkyl), wherein alkyl is as defined above.

The term "imino" refers to an ═ NR group where R can be selected from hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, and the like.

"optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs or does not. For example, "a heterocyclic group optionally substituted with an alkyl" means that an alkyl may, but need not, be present, and the description includes the case where the heterocyclic group is substituted with an alkyl and the heterocyclic group is not substituted with an alkyl.

"substituted" means that one or more, preferably up to 5, more preferably 1 to 3, hydrogen atoms in the group are independently substituted with a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and that the person skilled in the art is able to determine (experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups having free hydrogen may be unstable in combination with carbon atoms having unsaturated (e.g., olefinic) bonds.

"pharmaceutical composition" means a mixture containing one or more compounds described herein or a physiologically/pharmaceutically acceptable salt or prodrug thereof in admixture with other chemical components, as well as other components such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate administration to an organism, facilitate absorption of the active ingredient and exert biological activity.

"pharmaceutically acceptable salts" refers to salts of the compounds of the present invention which are safe and effective for use in the body of a mammal and which possess the requisite biological activity.

Synthesis of the Compounds of the invention

In order to achieve the purpose of the invention, the invention adopts the following technical scheme.

The preparation method of the compound shown in the general formula I or the salt thereof is as follows.

1) When R is¹is-C (O) R^aThe compounds of formula I can then be prepared from intermediates Ic by reaction with the corresponding acid chlorides according to the process of scheme 1.

Scheme 1

Synthesis of scheme 1:

under the high-temperature alkaline condition, corresponding bromides Ia and R containing boric acid pinacol ester⁵H, reacting to obtain an intermediate Ib, wherein a base reagent of the reaction is preferably potassium carbonate; reacting the intermediate Ib with 5-bromine- [1,2,4] under the action of a catalyst under the high-temperature alkaline condition]Triazolo [1,5-a]Carrying out Suzuki reaction on pyridine-2-amine to obtain an intermediate Ic; the intermediate Ic is reacted with the corresponding acid chloride under low temperature basic conditions to obtain the compound I, and the basic reagent of the reaction is preferably triethylamine.

2) When R is¹is-C (S) R^aThen, the method according to scheme 2 can be followed,

scheme 2

Synthesis of scheme 2:

and (3) reacting the intermediate Id with a Lawson reagent at a high temperature to obtain the compound shown in the general formula I.

3) When R is¹In the case of alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, the compounds of formula I can be prepared by conversion of intermediate Ic to iodo compound Ie and reaction with the corresponding amine according to scheme 3.

Scheme 3

Synthesis of scheme 3:

reacting the intermediate Ic with HI and NaNO under heating₂Reacting to obtain an intermediate Ie; the obtained intermediate Ie reacts with R under the action of a catalyst under the conditions of high-temperature alkaline microwave or high-temperature alkaline tube sealing¹NH₂The reaction is carried out to obtain the compound shown in the general formula I, the alkali reagent of the reaction is preferably sodium tert-butoxide, and the catalyst is preferably Pd₂(dba)₃、Xantphos。

4) When R is¹In the case of alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, the compounds of formula I can also be prepared according to the method of scheme 4.

Scheme 4

Synthesis of scheme 4:

under heating, 5-bromo- [1,2,4]]Triazolo [1,5-a]pyridin-2-Amines with HI, NaNO₂Reacting to obtain an intermediate If; the obtained intermediate Ie reacts with R under the action of a catalyst under the conditions of high-temperature alkaline microwave or high-temperature alkaline tube sealing¹NH₂The intermediate Ig is obtained by reaction of alkali reagent preferably sodium tert-butoxide and catalyst preferably Pd₂(dba)₃Xantphos; the obtained intermediate Ig reacts with an intermediate Ib under the action of a catalyst under the high-temperature alkaline condition to obtain a compound with a general formula I.

The reagents that provide basic conditions include organic bases including, but not limited to, triethylamine, N-diisopropylethylamine, N-butyllithium, lithium diisopropylamide, potassium acetate, sodium tert-butoxide, or potassium tert-butoxide, and inorganic bases including, but not limited to, sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, or cesium carbonate, sodium hydroxide, and lithium hydroxide.

Conditions that provide acidity include, but are not limited to, formic acid, acetic acid, hydrochloric acid, sulfuric acid, methanesulfonic acid.

Solvents used include, but are not limited to: acetic acid, methanol, ethanol, toluene, tetrahydrofuran, dichloromethane, dimethyl sulfoxide, 1, 4-dioxane, water or N, N-dimethylformamide.

Catalysts used include, but are not limited to: pd (dppf) Cl₂、Pd₂(dba)₃、Pd(OAc)₂。

R¹、R^1a、R^1b、R³、R^4a、R^4b、R⁵M1, n1 are as defined in formula (I).

Detailed Description

The present invention is further described below with reference to examples, which are not intended to limit the scope of the present invention.

The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) or/and Mass Spectrometry (MS). NMR shift at 10^-6The units in (ppm) are given. NMR was measured using a Brukerdps model 300 nuclear magnetic spectrometer using deuterated dimethyl sulfoxide (DMSO-d)₆) Deuterated chloroform (CDCl)₃) Deuterated methanol (CD)₃OD), internal standard Tetramethylsilane (TMS).

MS was measured using a 1100Series LC/MSD Trap (ESI) mass spectrometer (manufacturer: Agilent).

Liquid phase preparation lc3000 HPLC and lc6000 HPLC (manufacturer: Innovation Consumer) were used.

HPLC was carried out by using Shimadzu LC-20AD high pressure liquid chromatograph (Agilent TC-C18250X 4.6mm 5 μm column) and Shimadzu LC-2010AHT high pressure liquid chromatograph (Phenomenex C18250X 4.6mm 5 μm column).

Average inhibition rate of kinase and IC₅₀The values were measured using a multifunctional staining 3 microplate reader (Biotech, USA).

The thin layer chromatography silica gel plate is Qingdao ocean chemical GF254 silica gel plate, the specification of the silica gel plate used by Thin Layer Chromatography (TLC) is 0.15 mm-0.2 mm, and the specification of the thin layer chromatography separation and purification product is 0.4 mm-0.5 mm.

Column chromatography generally uses Qingdao marine silica gel 100-200 meshes and 200-300 meshes as a carrier.

Known starting materials of the present invention can be synthesized by or according to methods known in the art, or can be purchased from the companies such as cyber-mart, beijing coup, Sigma, carbofuran, yishiming, shanghai kaya, enokay, nanjing yashi, ann naiji chemical, and the like.

In the examples, the reaction can be carried out in an argon atmosphere or a nitrogen atmosphere, unless otherwise specified.

An argon atmosphere or nitrogen atmosphere means that the reaction flask is connected to a balloon of argon or nitrogen with a volume of about 1L.

The microwave reaction was carried out using a CEM Discover SP type microwave reactor.

In the examples, the solution means an aqueous solution unless otherwise specified.

In the examples, the reaction temperature is, unless otherwise specified, from 20 ℃ to 30 ℃ at room temperature.

The progress of the reaction in the examples was monitored by Thin Layer Chromatography (TLC) using a developing solvent system of: a: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, C: petroleum ether and ethyl acetate system, D: the volume ratio of acetone and solvent is adjusted according to the polarity of the compound.

The eluent system for column chromatography and the developing agent system for thin-layer chromatography used for purifying compounds comprise: a: dichloromethane and methanol system, B: petroleum ether, ethyl acetate and dichloromethane system, C: the volume ratio of the solvent in the petroleum ether and ethyl acetate system is adjusted according to the different polarities of the compounds, and a small amount of basic or acidic reagents such as triethylamine, acetic acid and the like can be added for adjustment.

Example 1: preparation of N- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) -2-oxopyrrolidine-3-carboxamide

Step 1: synthesis of 4- (4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -benzyl) thiomorpholine 1, 1-dioxide (intermediate 1A)

4-Bromomethylphenylboronic acid pinacol ester (50.00g,16.84mmol), 1-dioxide thiomorpholine (27.36g,20.24mmol) and potassium carbonate (27.92g,20.20mmol) were charged into a reaction flask, N-dimethylformamide (250 mL) was added, and the reaction was stirred at 80 ℃ for 4 hours. After cooling to room temperature, the reaction solution was poured into 1250mL of ice-water, stirred for 30 minutes, and filtered by suction to obtain the title product as a white solid (47.2 g, yield 79.7%).

Step 2: synthesis of 4- (4- (2-amino- [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide (intermediate 1B)

Reacting 5-bromo- [1,2,4]Triazolo [1,5-a]Pyridin-2-amine (30.00g,0.14mol) was charged into a 1L three-necked flask, and 4- [4- (4,4,5, 5-tetramethyl- [1,3,2 ] was sequentially added thereto]Dioxapentaborane-2-yl) -benzyl]-1, 1-dioxo-1-thiomorpholine (58.50g,0.15mol), dioxane (400mL), potassium carbonate (58.60g,0.42mol), water (100mL), Pd (dppf) Cl₂(5.78g,0.007 mol). The mixture was warmed to 90 ℃ under argon and the reaction was stirred for 16 hours. After the reaction was completed, it was cooled to room temperature, 400mL of dichloromethane was added, followed by washing twice with 800mL of water, the organic layer was concentrated to dryness, and the residue was purified by column chromatography (eluent: dichloromethane: methanol ═ 100:1) to give the title product as a pale yellow solid 27.2g, yield 54%.

And step 3: synthesis of N- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) -2-oxopyrrolidine-3-carboxamide (Compound 1)

4- (4- (2-amino- [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide (1.40g,3.9mmol) was charged to a 50mL reaction flask, and 7mL of methylene chloride and 1.63mL (11.7mmol) of triethylamine were sequentially added. 2-oxo-pyrrolidine-3-formyl chloride (prepared from 2-oxo-pyrrolidine-3-formic acid in Synlett,27(6),868-875 and 2016) (0.69g and 4.7mmol) is dissolved in 7mL of dichloromethane and added dropwise into the reaction solution, the temperature of the reaction solution is controlled to be 0-10 ℃ in the dropwise adding process, and the reaction solution reacts for 17 hours at room temperature after the dropwise adding is finished. After the reaction is finished, the mixture is washed once by 20mL of 10% hydrochloric acid aqueous solution, a brown solid is separated out, the mixture is filtered, the filtrate is kept stand for layering, an aqueous layer is reserved, the pH of the aqueous layer is adjusted to 10-11 by 10% NaOH aqueous solution, 20mL of dichloromethane is used for extraction, and the dichloromethane layer is concentrated to be dry to obtain a title product N- (5- (4- ((1, 1-dioxy thiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridine-2-yl) -2-oxo pyrrolidine-3-formamide, wherein the light yellow solid is 0.20g, and the yield is 11%.

ESI-MS[M+H](m/z):469.1.

¹H NMR(400MHz,DMSO)：δ10.99(s,1H),7.97(m,2H),7.87(m,1H),7.70(m,2H),7.51(m,2H),7.30(m,1H),3.75(s,2H),3.13(m,4H),2.91(m,4H),2.47(s,1H),2.12(m,3H),1.15(m,1H).

Example 2: preparation of N- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) pyrrolidine-2-carboxamide

The title compound was obtained in the same manner as in example 1, except that prolyl chloride (prepared from Boc-L-proline as the starting material in reference Tetrahedron,65(25), 4841-4845; 2009) was used instead of 2-oxo-pyrrolidine-3-carbonyl chloride.

ESI-MS[M+H](m/z):455.1.

¹H NMR(300MHz,DMSO)：δ10.38(s,1H),7.99(m,2H),7.90(m,1H),7.73(m,2H),7.53(m,2H),7.35(m,1H),3.89(m,1H),3.73(s,2H),3.40(m,2H),3.13(m,4H),2.94(m,4H),1.74(m,4H).

Example 3: preparation of N- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) -1-methylazetidine-3-carboxamide

The title compound was obtained in the same manner as the preparation of example 1, except that 1-methyl-azetidine-3-carbonyl chloride (prepared starting from 3-azetidinecarboxylic acid in reference WO 2009032326) was used instead of 2-oxo-pyrrolidine-3-carbonyl chloride.

ESI-MS[M+H](m/z):455.1.

¹H NMR(300MHz,DMSO)：δ10.45(s,1H),8.01(m,2H),7.88(m,1H),7.72(m,2H),7.49(m,2H),7.36(m,1H),3.73(s,2H),3.31-3.72(m,4H),3.39(m,1H),3.15(m,4H),2.95(m,4H),2.33(s,3H).

Example 4: preparation of N- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) -1H-pyrazole-5-carboxamide

The title compound was obtained in the same manner as the preparation of example 1 except that 1H-pyrazole-5-carbonyl chloride was used instead of 2-oxo-pyrrolidine-3-carbonyl chloride.

ESI-MS[M+H](m/z):452.1.

¹H NMR(300MHz,DMSO)：δ10.60(s,1H),8.02-8.14(m,2H),7.90(m,1H),7.75(m,2H),7.55(m,2H),7.35(m,1H),7.22(m,1H),3.79(s,2H),3.15(m,4H),2.93(m,4H).

Example 5: preparation of N- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) thiophene-2-carboxamide

The title compound was obtained in the same manner as the preparation of example 1 except that thiophene-2-carbonyl chloride was used instead of 2-oxo-pyrrolidine-3-carbonyl chloride.

ESI-MS[M+H](m/z):468.2.

¹H NMR(300MHz,DMSO)：δ10.60(s,1H),8.02-8.14(m,3H),7.90(m,1H),7.75(m,2H),7.55(m,2H),7.35(m,1H),7.22(m,1H),4.71(s,2H),3.15(m,4H),2.93(m,4H).

Example 6: preparation of N- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) -1H-imidazole-2-carboxamide

The title compound was obtained in the same manner as the preparation of example 1, except that imidazole-2-carbonyl chloride (prepared starting from imidazole-2-carboxylic acid, reference CN 102153515) was used instead of 2-oxo-pyrrolidine-3-carbonyl chloride.

ESI-MS[M+H](m/z):452.1.

¹H NMR(300MHz,DMSO)：δ10.55(s,1H),8.09(m,2H),7.87(m,1H),7.75(m,2H),7.55(m,2H),7.35(m,1H),7.25(m,1H),7.15(m,1H),6.98(m,1H),3.79(s,2H),3.16(m,4H),2.99(m,4H).

Example 7: preparation of N- (5- (4- ((2, 2-dioxo-2-thia-6-azaspiro [3.3] heptan-6-yl) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide

Step 1: preparation of N- (5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (intermediate 7A)

50mL of pyridine was added to a reaction flask, cyclopropanecarbonyl chloride (2.9g,28mmol) was added dropwise to the reaction flask at-50 ℃ and stirred for 10 minutes, and then 5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-amine (5g,23mmol) was added to the reaction flask and stirred at room temperature overnight. After the reaction was completed, 100mL of petroleum ether was added to the reaction solution, followed by suction filtration, and the filter cake was washed twice with 100mL of petroleum ether and then once with 100mL of water to obtain the title product as a grayish solid (4.5 g).

Step 2: preparation of N- (5- (4- (hydroxymethyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (intermediate 7B)

N- (5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (2.8g,10mmol), 4-hydroxymethylphenylboronic acid (1.5g,10mmol), 1' -bisdiphenylphosphinoferrocene palladium dichloride (730mg,1mmol), and potassium carbonate (2.8g,20mmol) were sequentially added to a single vial, and then 50mL of 1, 4-dioxane and 10mL of water were added thereto. The reaction was heated to 90 ℃ and stirred for 16 hours under argon. Then, the reaction solution was cooled to room temperature, extracted with ethyl acetate, and the organic layer was separated, dried, and concentrated. The residue was purified by column chromatography (eluent: dichloromethane: methanol ═ 25: 1) to give the title product in the form of a yellow solid 2.1g with a yield of 68%.

And step 3: preparation of N- (5- (4- (bromomethyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (intermediate 7C)

N- (5- (4- (hydroxymethyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (200mg,0.65mmol), triphenylphosphine (260mg,1mmol) and carbon tetrabromide (330mg,1mmol) were placed in a single vial, and 5mL of dichloromethane was added, followed by stirring at room temperature for 15 hours. The reaction solution was stirred with silica gel and directly purified by column chromatography (eluent: dichloromethane: methanol ═ 30: 1) to give a mixture of [5- (4-bromomethyl-phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl ] -cyclopropanecarboxamide and triphenylphosphine oxide as a white solid (150 mg).

And 4, step 4: preparation of N- (5- (4- ((2, 2-dioxo-2-thia-6-azaspiro [3.3] heptan-6-yl) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (Compound 7)

100mg of crude N- (5- (4- (bromomethyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide, 2-thio-6-azaspiro [3.3] heptane-2, 2-dioxide (60mg,0.4mmol) and potassium carbonate (120mg,0.8mmol) were added to a single-necked flask, N-dimethylformamide (2mL) was added thereto, and the mixture was stirred at room temperature for 16 hours. To the reaction solution was added 50mL of ethyl acetate, and the mixture was washed three times with a saturated aqueous solution of sodium chloride, and the organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by thin layer chromatography (developing solvent: chloroform: methanol ═ 20:1) to give the title product as a white solid, 42 mg.

MS[M+H](m/z):438.2.

¹H NMR(300MHz,DMSO)：δ10.50(s,1H),8.11(m,2H),7.83(m,1H),7.75(m,2H),7.51(m,2H),7.35(m,1H),3.73(s,2H),3.35(m,4H),3.21(m,4H),2.09(m,1H),1.12(m,2H),0.85(m,2H).

Example 8: preparation of N- (5- (4- ((1, 1-dioxythiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarbothioamide

Step 1: preparation of N- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (intermediate 8A)

4- (4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -benzyl) thiomorpholine 1, 1-dioxide (intermediate 1A) (20.18g,53mmol), N- (5-bromo- [1,2, 4-mmol)]Triazolo [1,5-a]Pyridin-2-yl) cyclopropanecarboxamide (intermediate 7A) (16.45g,59mmol), Pd (dppf) Cl₂(3.91g,5mmol)、K₂CO₃(22.06g,160mmol) was added to a 250mL three-necked flask, followed by 1, 4-dioxane (100mL) and water (25mL), and the mixture was heated to 90 ℃ under argon atmosphere for 12 hours. After completion of the reaction, it was cooled to room temperature, 300mL of dichloromethane was added to the reaction solution, which was then washed twice with 500mL of water, the organic phase was dried over anhydrous sodium sulfate, the organic phase was concentrated to dryness, and the residue was purified by column chromatography (eluent: dichloromethane: methanol ═ 50: 1) to give the title product as a pale yellow solid, 11 g.

Step 2: preparation of N- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarbothioamide (Compound 8)

N- (5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) cyclopropanecarboxamide (1.00g,2.4mmol) was added to a 50mL single-neck flask, to which were added in this order toluene 20mL, Lawson's reagent (0.48g,1.2 mmol). The mixture was refluxed for 7 hours under an argon atmosphere. After the reaction is finished, cooling to room temperature, carrying out suction filtration, and concentrating the filtrate under reduced pressure to dryness. The residue was purified by column chromatography (eluent: dichloromethane: methanol ═ 50: 1) to give the title product as a yellow solid, 0.8 g.

ESI-MS[M+H](m/z):442.1.

¹H NMR(300MHz,DMSO)：δ10.03(s,1H),8.08(m,2H),7.85(m,1H),7.72(m,2H),7.54(m,2H),7.35(m,1H),3.79(s,2H),3.20(m,4H),2.95(m,4H),2.58(m,1H),0.93(m,2H),0.61(m,2H).

Example 9: preparation of 4- (4- (2- ((1-methyl-1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

Step 1: synthesis of 4- (4- (2-iodo- [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide (intermediate 9A)

HI (8mL) and DMSO (40mL) were added to a 250mL reaction flask. To this was slowly added, while stirring at room temperature, a solution of 4- (4- (2-amino- [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide (intermediate 1B) (4.00g,11mmol) and sodium nitrite (3.08g,44mmol) in dimethylsulfoxide (60 mL). The remaining insoluble sodium nitrite was added to 20mL of water, stirred to full solubility and then added to the reaction flask. Then, the reaction solution was heated to 35 ℃ and stirred to react for 6 hours. After the reaction was completed, it was cooled to room temperature, the pH was adjusted to 9 with an aqueous potassium carbonate solution, then 400mL of ethyl acetate was added for extraction, and the organic phase was washed once with 400mL of a 5% aqueous sodium thiosulfate solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title product as a pale yellow solid 3.0g, yield 57%.

Step 2: synthesis of 4- (4- (2- ((1-methyl-1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide (Compound 9)

Mixing 4- (4- (2-iodine- [1,2, 4))]Triazolo [1,5-a]Pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide (50mg, 0.1)3mmol), 1-methyl-4-aminopyrazole (19mg,0.20mmol), Pd₂(dba)₃(16mg,0.01mmol), Xantphos (8mg,0.01mmol) and sodium tert-butoxide (63mg,0.65mmol) were added sequentially to the reaction flask, followed by dioxane 3mL and argon blanketing for 3 minutes with stirring. Heating to 160 ℃ under microwave condition to react for 30 minutes. After the reaction was completed, the reaction mixture was cooled to room temperature, filtered through 3g of celite, the filter cake was washed with 10mL of methanol, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by column chromatography (eluent: dichloromethane: methanol ═ 50: 1) to give the title product as a white solid, 7mg, yield: 12% and the purity is 96%.

MS[M+H](m/z):438.1.

¹H NMR(300MHz,DMSO)：δ9.16(s,1H),7.97(m,2H),7.63(m,1H),7.44(m,4H),7.35(m,1H),7.09(m,1H),3.72(m,5H),3.10(m,4H),2.88(m,4H).

Example 10: preparation of 4- (4- (2- (oxazol-2-ylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 2-aminooxazole instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):425.1.

¹H NMR(300MHz,DMSO)：δ9.16(s,1H),7.97(m,2H),7.66(m,1H),7.50(m,3H),7.35(m,2H),7.20(m,1H),3.75(s,2H),3.13(m,4H),2.85(m,4H).

Example 11: preparation of 4- (4- (2- (thiazol-2-ylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 2-aminothiazole instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):441.1.

¹H NMR(300MHz,DMSO)：δ9.10(s,1H),7.97(m,2H),7.68(m,1H),7.53(m,2H),7.39(m,2H),7.27(m,1H),6.94(m,1H),3.73(s,2H),3.10(m,4H),2.89(m,4H).

Example 12: preparation of 4- (4- (2- (cyclopropylamine) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using cyclopropylamine instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):398.2.

¹H NMR(300MHz,DMSO)：δ8.03(m,2H),7.78(m,1H),7.60(m,2H),7.49(m,2H),7.35(m,1H),6.08(s,1H),3.75(s,2H),2.43(m,1H),0.76(m,4H).

Example 13: preparation of 4- (4- (2- ((1, 4-dimethyl-1H-pyrazol-5-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 1, 4-dimethyl-1H-pyrazol-5-amine instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):452.1.

¹H NMR(300MHz,DMSO)：δ9.25(s,1H),8.03(m,2H),7.65(m,1H),7.44(m,3H),7.39(m,1H),7.22(m,1H),3.72(m,5H),3.10(m,4H),2.88(m,4H),2.10(s,3H).

Example 14: preparation of 4- (4- (2- (pyridin-4-ylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 4-aminopyridine instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):435.5.

¹H NMR(300MHz,DMSO):δ10.24(s,1H),8.32(m,2H),8.02(m,2H),7.64(m,6H),7.27(m,1H),3.81(s,2H),3.16(m,4H),2.94(m,4H).

Example 15: preparation of 4- (4- (2- ((1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 4-aminopyrazole in place of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):424.1.

¹H NMR(300MHz,DMSO):δ9.10(s,1H),8.00(m,2H),7.60(m,1H),7.50(m,4H),7.30(m,1H),7.03(m,1H),5.18(s,1H),3.69(s,2H),3.09(m,4H),2.85(m,4H).

Example 16: preparation of 4- (4- (2- ((1-isopropyl-1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 4-amino-1-isopropyl-1H-pyrazole in place of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):466.1.

¹H NMR(300MHz,DMSO):δ9.22(s,1H),8.06(m,2H),7.78(m,1H),7.54(m,4H),7.48(m,1H),7.14(m,1H),4.41(m,1H),3.80(m,1H),3.14(m,4H),2.94(m,4H),1.39(m,6H).

Example 17: preparation of 1- (4- ((5- (4- ((1, 1-dioxothiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) amino) -1H-pyrazol-1-yl) ethan-1-one

The title compound was obtained in the same manner as the preparation of example 9 except for using 1- (4-amino-pyrazol-1-yl) -ethanone instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):466.2.

¹H NMR(300MHz,DMSO):δ9.08(s,1H),8.00(m,2H),7.66(m,1H),7.49(m,4H),7.36(m,1H),7.11(m,1H),3.68(s,2H),3.10(m,4H),2.88(m,4H),2.79(s,3H).

Example 18: preparation of 4- (4- (2- ((2-fluoro-4, 5-dimethoxyphenyl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 2-fluoro-4, 5-dimethoxyaniline in place of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):512.2.

¹H NMR(300MHz,DMSO):δ9.13(s,1H),8.04(m,2H),7.75(m,1H),7.54(m,3H),7.48(m,1H),7.06(m,1H),6.83(m,1H),3.79(s,2H),3.95(s,3H),3.84(s,3H),3.16(m,4H),2.95(m,4H).

Example 19: preparation of 4- (4- (2- ((2-fluoro-4-methoxyphenyl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 2-fluoro-4-methoxyaniline in place of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):482.1.

¹H NMR(300MHz,DMSO):δ9.18(s,1H),8.07(m,2H),7.78(m,1H),7.55(m,3H),7.44(m,1H),7.08(m,2H),6.69(m,1H),3.76(s,2H),3.89(s,3H),3.18(m,4H),2.97(m,4H).

Example 20: preparation of 3- ((5- (4- ((1, 1-dioxythiomorpholine) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-yl) amino) benzenesulfonamide

The title compound was obtained in the same manner as the preparation of example 9 except that m-aminobenzenesulfonamide was used instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):513.2.

¹H NMR(300MHz,DMSO):δ9.15(s,1H),8.09(m,2H),7.81(m,1H),7.52(m,8H),5.18(s,2H),3.73(s,2H),3.18(m,4H),2.97(m,4H).

Example 21: preparation of 4- (4- (2- ((1-cyclopropyl-1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 4-amino-1-cyclopropyl-1H-pyrazole in place of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):464.1.

¹H NMR(300MHz,DMSO):δ9.23(s,1H),8.08(m,2H),7.78(m,1H),7.50(m,4H),7.46(m,1H),7.15(m,1H),3.78(s,2H),3.32(m,1H),3.14(m,4H),2.94(m,4H),1.15(m,4H).

Example 22: preparation of 4- (4- (2- ((1-methyl-1H-indazol-6-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as in the preparation of example 9 except for using 1-methyl-6-amino-1H-indazole instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):488.5.

¹H NMR(300MHz,DMSO):δ9.13(s,1H),8.15(m,1H),8.09(m,2H),7.80(m,2H),7.54(m,4H),7.45(m,1H),7.10(m,2H),4.13(s,3H),3.73(s,2H),3.16(m,4H),2.97(m,4H).

Example 23: preparation of 4- (4- (2- (5-methyl-4, 5,6, 7-tetrahydropyrazolo [1,5-a ] pyrazin-2-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except that (5-methyl-4, 5,6, 7-tetrahydropyrazolo [1,5-a ] pyrazin-2-amine was used instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):493.2.

¹H NMR(300MHz,DMSO):δ9.65(s,1H),8.02(m,2H),7.66(m,5H),7.15(m,1H),6.32(m,1H),3.93(m,2H),3.79(s,1H),3.52(s,2H),3.15(s,5H),2.94(s,5H),2.80(m,2H),2.50(m,2H).

Example 24: preparation of 4- (4- (2- ((1- (1-methylazetidin-3-yl) -1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 1- (1-methylazetidin-3-yl) -1H-pyrazol-4-amine instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):493.2.

¹H NMR(300MHz,DMSO):δ9.22(s,1H),8.09(m,2H),7.73(m,1H),7.52(m,4H),7.46(m,1H),7.15(m,1H),4.40(m,1H),3.79(m,4H),3.71(s,2H),2.28(s,3H).

Example 25: preparation of 4- (4- (2- ((1- (1- (ethylsulfonyl) -1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 1- (ethylsulfonyl) -1H-pyrazol-4-amine instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):516.2.

¹H NMR(300MHz,DMSO):δ9.13(s,1H),8.02(m,2H),7.53(m,5H),7.39(m,1H),7.13(m,1H),3.68(s,2H),3.10(m,7H),2.88(m,4H),1.35(m,3H).

Example 26: preparation of 4- (4- (2- (cyclopentylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using cyclopentylamine in place of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):426.2.

¹H NMR(300MHz,DMSO):δ8.03(m,2H),7.66(m,1H),7.50(m,3H),7.35(m,1H),6.13(s,1H),3.75(s,2H),3.13(m,4H),3.02(m,1H),2.85(m,4H),1.83(m,4H),1.65(m,4H).

Example 27: preparation of 4- (4- (2- (isoxazol-4-ylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using isoxazol-4-amine in place of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):425.2.

¹H NMR(300MHz,DMSO)：δ9.20(s,1H),8.01(m,2H),7.66(m,1H),7.50(m,3H),7.35(m,1H),6.37(m,1H),3.71(s,2H),3.13(m,4H),2.85(m,4H).

Example 28: preparation of 4- (4- (2- (isothiazol-4-ylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using isothiazol-4-amine in place of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):441.2.

¹H NMR(300MHz,DMSO)：δ9.27(s,1H),8.01(m,2H),7.62(m,1H),7.50(m,4H),7.37(m,1H),7.25(m,1H),3.71(s,2H),3.12(m,4H),2.83(m,4H).

Example 29: preparation of 4- (4- (2- ((1-methylpiperidin-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 1-methylpiperidin-4-amine instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):455.2.

¹H NMR(300MHz,DMSO):δ8.06(m,2H),7.82(m,1H),7.47(m,3H),7.38(m,1H),5.95(s,1H),3.66(s,2H),3.51(m,4H),2.92(m,4H),2.63(m,4H),2.51(m,2H),2.41(m,2H),2.18(s,3H),1.81(m,2H),1.56(m,2H).

Example 30: preparation of 4- (4- (2- (cyclohexylamino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except that cyclohexylamine was used instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):440.2.

¹H NMR(300MHz,DMSO):δ8.02(m,2H),7.66(m,1H),7.50(m,3H),7.35(m,1H),6.13(s,1H),3.69(s,2H),3.13(m,4H),3.06(m,1H),2.85(m,4H),1.98(m,4H),1.73(m,5H),1.39(m,2H).

Example 31: preparation of 4- (4- (2- ((1- (2,2, 2-trifluoroethyl) -1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 1- (2,2, 2-trifluoroethyl) -1H-pyrazol-4-amine instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):506.2.

¹H NMR(300MHz,DMSO):δ8.18(s,1H),8.06(m,2H),7.82(m,2H),7.50(m,3H),7.38(m,1H),7.18(m,1H),4.25(m,2H),3.66(s,2H),3.51(m,4H),2.92(m,4H).

Example 32: preparation of 4- (4- (2- ((1- (difluoromethyl) -1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 1- (difluoromethyl) -1H-pyrazol-4-amine instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):474.2.

¹H NMR(300MHz,DMSO):δ8.18(s,1H),8.06(m,2H),7.82(m,2H),7.50(m,3H),7.38(m,1H),7.36(m,1H),7.15(m,1H),3.66(s,2H),3.51(m,4H),2.92(m,4H).

Example 33: preparation of 4- (4- (2- ((1- (2-methoxyethyl) -1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 1- (2-methoxyethyl) -1H-pyrazol-4-amine in place of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):482.2.

¹H NMR(300MHz,DMSO):δ8.18(s,1H),8.06(m,2H),7.82(m,2H),7.47(m,3H),7.38(m,1H),7.18(m,1H),3.85(m,4H),3.66(s,2H),3.51(m,4H),3.23(s,3H),2.92(m,4H).

Example 34: preparation of 4- (4- (2- ((1- (2-hydroxyethyl) -1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 1- (2-hydroxyethyl) -1H-pyrazol-4-amine instead of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):468.2.

¹H NMR(300MHz,DMSO):δ8.18(s,1H),8.06(m,2H),7.82(m,2H),7.47(m,3H),7.38(m,1H),7.18(m,1H),4.96(m,1H),3.92(m,2H),3.66(m,4H),3.51(m,4H),2.91(m,4H).

Example 35: preparation of N- (1-methyl-1H-pyrazol-4-yl) -5- (4- (morpholinomethyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

Step 1: synthesis of 5-bromo-2-iodo- [1,2,4] triazolo [1,5-a ] pyridine (intermediate 35A)

The title compound was obtained in the same manner as the preparation of intermediate 9A except that 5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-amine was used instead of 4- (4- (2-amino- [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide.

Step 2: synthesis of 5-bromo-N- (1-methyl-1H-pyrazol-4-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine (intermediate 35B)

Mixing 5-bromo-2-iodo- [1,2, 4%]Triazolo [1,5-a]Pyridine (500mg,1.6mmol), 1-methyl-4-aminopyrazole (190mg,2.0mmol), Pd₂(dba)₃(160mg,0.1mmol), Xantphos (80mg,0.1mmol) and sodium tert-butoxide (630mg,6.5mmol) were added sequentially to the reaction flask, followed by addition of 20mL dioxane and 3 minutes under argon with stirring. Heating to 160 ℃ under microwave condition to react for 30 minutes. After the reaction was completed, the reaction mixture was cooled to room temperature, filtered through 3g of celite, the filter cake was washed with 10mL of methanol, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by column chromatography (eluent: dichloromethane: methanol ═ 50: 1) to give the title product as a white solid 230mg, yield: 51 percent.

And step 3: synthesis of 4- (4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) morpholine (intermediate 35C)

4-Bromomethylphenylboronic acid pinacol ester (5.00g,1.68mmol), morpholine (1.74g,2.00mmol) and potassium carbonate (2.80g,2.00mmol) were charged into a reaction flask, and N, N-dimethylformamide (50 mL) was added thereto, followed by stirring at 80 ℃ for 4 hours. After cooling to room temperature, the reaction solution was poured into 250mL of ice water, stirred for 30 minutes, and filtered by suction to give the title product as a white solid (3.7 g) with a yield of 72.6%.

And 4, step 4: synthesis of N- (1-methyl-1H-pyrazol-4-yl) -5- (4- (morpholinomethyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine (Compound 35)

Reacting 5-bromo-N- (1-methyl-1H-pyrazol-4-yl) - [1,2, 4%]Triazolo [1,5-a]Pyridin-2-amine (100mg,0.34mmol)Into a three-necked flask, 4- (4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) morpholine (121mg,0.40mol), dioxane 10mL, potassium carbonate (196mg,1.42mmol), water 2mL, Pd (dppf) Cl₂(51mg,0.07 mmol). The mixture was warmed to 90 ℃ under argon and the reaction was stirred for 16 hours. After the reaction was completed, it was cooled to room temperature, 20mL of dichloromethane was added, followed by washing twice with 20mL of water, the organic layer was concentrated to dryness, and the residue was purified by column chromatography (eluent: dichloromethane: methanol ═ 20:1) to give the title product as a white solid 23mg, yield 17%.

MS[M+H](m/z):390.1.

¹H NMR(300MHz,DMSO)：δ9.20(s,1H),7.99(m,2H),7.68(m,1H),7.50(m,4H),7.39(m,1H),7.11(m,1H),3.76(s,3H),3.59(m,6H),2.40(m,4H).

Example 36: preparation of 5- (4- ((4, 4-difluoropiperidin-1-yl) methyl) phenyl) -N- (1-methyl-1H-pyrazol-4-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

The title compound was obtained in the same manner as the preparation of example 35 except that 4, 4-difluoropiperidine was used instead of morpholine.

MS[M+H](m/z):424.2.

¹H NMR(300MHz,DMSO):δ9.21(s,1H),8.02(m,2H),7.52(m,6H),7.14(m,1H),3.76(s,3H),3.65(s,2H),2.53(m,4H),2.00(m,4H).

Example 37: preparation of N- (1-methyl-1H-pyrazol-4-yl) -5- (4- (piperidin-1-ylmethyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

The title compound was obtained in the same manner as the preparation of example 35 except that piperidine was used instead of morpholine.

MS[M+H](m/z):388.3.

¹H NMR(300MHz,DMSO):δ9.17(s,1H),7.99(m,2H),7.66(m,1H),7.48(m,4H),7.35(m,1H),7.10(m,1H),3.73(m,5H),2.50(m,4H),1.55(m,4H),1.43(m,2H).

Example 38: preparation of N- (1-methyl-1H-pyrazol-4-yl) -5- (4- (pyrrolidin-1-ylmethyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

The title compound was obtained in the same manner as the preparation of example 35 except that pyrrolidine was used instead of morpholine.

MS[M+H](m/z):374.2.

¹H NMR(300MHz,DMSO):δ9.16(s,1H),7.95(m,2H),7.66(m,1H),7.48(m,4H),7.35(m,1H),7.10(m,1H),3.69(m,2H),3.05(m,2H),2.49(m,2H),1.74(m,4H).

Example 39: preparation of N- (1-methyl-1H-pyrazol-4-yl) -5- (4- ((6-methyl-2, 6-diazaspiro [3.3] hept-2-yl) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

The title compound was obtained in the same manner as the preparation of example 35 except that 2-methyl-2, 6-diazaspiro [3.3] heptane oxalate was used instead of morpholine.

MS[M+H](m/z):415.2.

¹H NMR(300MHz,DMSO):δ9.18(s,1H),7.98(m,2H),7.66(m,1H),7.48(m,4H),7.37(m,1H),7.09(m,1H),3.73(m,5H),3.50-3.28(m,8H),2.23(s,3H).

Example 40: preparation of N- (1-methyl-1H-pyrazol-4-yl) -5- (4- ((5-methyl-hexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

The title compound was obtained in the same manner as the preparation of example 35 except that 2-methyloctahydropyrrolo [3,4-c ] pyrrole was used instead of morpholine.

MS[M+H](m/z):429.2.

¹H NMR(300MHz,DMSO):δ9.20(s,1H),8.00(m,2H),7.65(m,1H),7.46(m,4H),7.37(m,1H),7.09(m,1H),3.69(m,5H),3.39(m,2H),2.53(m,4H),2.26(m,4H),2.09(s,3H).

Example 41: preparation of N- (1-methyl-1H-pyrazol-4-yl) -5- (4- ((3-methyl-3, 8-diazabicyclo [3.2.1] octan-8-yl) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

The title compound was obtained in the same manner as the preparation of example 35 except that 3-methyl-3, 8-diazabicyclo [3.2.1] octane dihydrochloride was used instead of morpholine.

MS[M+H](m/z):429.2.

¹H NMR(300MHz,DMSO):δ9.18(s,1H),7.95(m,2H),7.66(m,1H),7.50(m,4H),7.35(m,1H),7.10(m,1H),3.73(m,5H),2.83(m,2H),2.51(m,2H),2.29(m,2H),2.15(m,5H),1.53(m,4H).

Example 42: preparation of 4- (4- (2- ((4, 4-dimethyl-4, 5-dihydrooxazol-2-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) thiomorpholine 1, 1-dioxide

The title compound was obtained in the same manner as the preparation of example 9 except for using 4, 4-dimethyl-4, 5-dihydrooxazol-2-amine in place of 1-methyl-4-aminopyrazole.

MS[M+H](m/z):4552.

¹H NMR(300MHz,CDCl₃):δ8.66(s,1H),8.02(m,2H),7.51(m,2H),7.44(m,2H),7.02(m,1H),4.19(s,2H),3.74(m,2H),3.06(m,8H),1.50(s,6H).

Example 43: preparation of 1- (4- (4- (2- ((1-methyl-1H-pyrazol-4-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzyl) piperazin-1-yl) ethan-1-one

The title compound was obtained in the same manner as the preparation of example 35 except that 1- (piperazin-1-yl) eth-1-one was used instead of morpholine.

MS[M+H](m/z):431.2.

¹H NMR(300MHz,DMSO):δ9.16(s,1H),8.01(m,2H),7.64(m,1H),7.50(m,4H),7.36(m,1H),7.12(m,1H),3.70(m,5H),3.31(m,4H),2.81(m,4H),2.15(s,3H).

Example 44: preparation of N- (1-methyl-1H-pyrazol-4-yl) -5- (4- ((4- (methylsulfonyl) piperazin-1-yl) methyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

The title compound was obtained in the same manner as the preparation of example 35 except that 1- (methylsulfonyl) piperazine was used instead of morpholine.

MS[M+H](m/z):467.2.

¹H NMR(300MHz,DMSO):δ9.18(s,1H),8.00(m,2H),7.64(m,1H),7.50(m,4H),7.35(m,1H),7.10(m,1H),3.69(m,5H),2.88(m,7H),2.69(m,4H).

Biological evaluation

Test example 1: in vitro kinase Activity

Experimental materials: JAK1, JAK2, JAK3 and Tyk2 kinases, substrates for kinases GFP-STAT1, ATPLATANTHASCREEN^TMTb-anti-pSTAT1, EDTA, TR-FRET Dilution Buffer for kinase reaction, and Filgotinib as a control were obtained from the following materials as shown in Table 1.

TABLE 1

Preparing a medicine: the compound of the present invention was dissolved in DMSO solvent to prepare a 10mM stock solution. The final compound reaction maximum concentration is 10uM, 3 times of dilution, 10 concentration gradient, each concentration gradient is provided with 3 multiple holes.

The experimental method comprises the following steps: adding JAK1(500ng/mL), JAK2(15ng/mL), JAK3(250ng/mL) and Tyk2(200ng/mL) kinases into 384 reaction plates containing the compounds, and incubating for 15 minutes in an incubator at 25 ℃; then, 4. mu.L of the substrate mixture (20. mu.M ATP and 0.1. mu.M GFP-STAT1) was added to a 384 reaction plate containing JAK kinase and the compound, and the reaction was carried out in an incubator at 25 ℃ for 1 hour; mu.L of the antibody mixture (10mM EDTA and 2nM antibody) was added to the 384 reaction plate and reacted in a constant temperature incubator at 25 ℃ for 1 hour; the 384 reaction plates were taken out and read on an Envision multifunctional plate reader.

IC of the compound was obtained using the following non-linear fit equation₅₀(median inhibitory concentration):

Y＝Bottom+(Top-Bottom)/(1+10^((LogIC50-X)*HillSlope))

x: log value of compound concentration

Y: emissivity (emision Ratio)

Bottom: minimum, Top: highest value, Hill Slope: slope of

The inhibitory activity of the compounds of the present invention against JAK1, JAK2, JAK3 and Tyk2 is shown in table 2 below.

TABLE 2 IC for in vitro JAK kinase inhibitory Activity of Compounds of the invention₅₀

Remarking: IC (integrated circuit)₅₀Values from 0 to 10nM are marked A; 10-100nM is marked B; greater than 100 is labeled C; NT stands for not tested.

As is clear from table 2, the compound of the present invention has excellent inhibitory activity against four kinases, JAK1, JAK2, JAK3 and Tyk2, wherein the inhibitory activity against JAK1 is 5 times or more higher than that against Filgotinb.

From the test results, it is clear that the compound of the invention has good in vitro anti-JAK kinase activity, and is equivalent to or superior to clinical II stage antirheumatic drug Filgotinib.

Test example 2: type II collagen-induced DBA/1J mouse arthritis model (CIA)

The CIA model is an animal model widely used for studying the activity of drugs for treating rheumatoid arthritis in humans.

Animals: DBA/1J mice, sex: male, number: 30, body weight: 14-16g, week age range: 6-7 weeks old, purchased from Witonglihua laboratory animal technology, Inc., Beijing, SPF grade, animal production license number: SCXK (Jing) 2012-0001, issuing unit: the scientific and technical committee of Beijing.

Grouping: model control group, administration (30mg/Kg) group (Compound of example 35)

An appropriate amount of the Bovine Type II Collage (from Chondrex. Inc.) was taken, dissolved in 0.05M acetic acid (4mg collagen/mL) and refrigerated overnight at4 ℃. The mixture was well emulsified with an equal amount of Freund's complete adjuvant in an ice bath, and each DBA/1J mouse was injected with 0.1mL (200. mu.g collagen) of the emulsion from the 2cm of the caudal root until the needle tip was inserted 0.5cm from the caudal root. Starting clinical symptom observation and arthritis scoring after 4 weeks of model building, sorting scores of all animals with symptoms, randomly grouping layer by layer, and dividing into 2 groups, namely a model control group (oral gavage is given with 0.5% CMC as a solvent) and an administration group (oral gavage is given with 30mg/kg of the compound of the embodiment 35); 2 times/day for 14 days; arthritis scores and weight changes were observed every 3 days after dosing.

Arthritis index score: the arthritis index was scored according to Wood's arthritis scoring criteria, as follows.

0 minute: is normal

1 minute: red swelling involving 1 finger joint

And 2, dividing: the mild red swelling of more than 2 finger joints or the entire paw

And 3, dividing: severe red and swollen foot and paw

And 4, dividing: severe redness and swelling of the paw, stiffness of the joints, and lack of elasticity.

Lesions were divided between 0 and 4 for each of the 4 paws and the total integral of the limb was calculated. The integrals (arthritis index) at different times were compared. The results are shown in table 3 below.

TABLE 3 Effect of the Compounds of example 35 of the invention on the arthritis score in CIA model animals

Remarking: p <0.05 compared to model control group.

And (4) conclusion: as can be seen from the above table 3, the compound of the present invention significantly reduces the arthritis score of CIA model animals, which indicates that the compound of the present invention has a significant improvement effect on the arthritis symptoms of CIA mice.

Claims

1. A compound, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

n- (1-methyl-1H-pyrazol-4-yl) -5- (4- (morpholinomethyl) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine.

2. A pharmaceutical composition comprising a compound according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, diluent or excipient.

3. Use of a compound according to claim 1 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 2 in the preparation of a Janus kinase inhibitor.

4. Use of a compound according to claim 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 2, in the manufacture of a medicament for the treatment of a disease associated with Janus kinase activity, wherein the disease is selected from inflammation, an autoimmune disease, or cancer.

5. The use of claim 4, wherein the inflammation is arthritis.

6. Use according to claim 5, wherein the arthritis is rheumatoid arthritis, psoriatic arthritis.

7. The use of claim 4, wherein the inflammation is inflammatory enteritis, uveitis, or psoriasis.

8. The use of claim 4, wherein the autoimmune disease is multiple sclerosis or lupus.

9. The use of claim 4, wherein the cancer is a solid tumor.

10. The use of claim 4, wherein the cancer is breast cancer, cervical cancer, colon cancer, lung cancer, stomach cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor, ovarian tumor, peritoneal tumor, melanoma, glioma, glioblastoma, papillary renal tumor, head and neck tumor, leukemia, lymphoma, myeloma, or non-small cell lung cancer.

11. The use of claim 4, wherein the cancer is hepatocellular carcinoma.