Heteroaryl compounds as RIP2 kinase inhibitors, compositions thereof, and uses thereof

Abstract

The present disclosure provides heterocyclic compounds having RIP2 kinase inhibitory activity, pharmaceutical compositions containing the same, methods using the same and applications thereof. The present disclosure provides compounds of formula (I) as inhibitors of RIP2 kinase. These compounds can be used to prevent and / or treat diseases and / or conditions associated with RIP2 kinase. [Formula 1] JPEG2024507747000087.jpg50166

Description

Technical Field

[0001] (Cross - reference to related applications) This application claims the benefit of Chinese Patent Application No. 202110272207.4 filed on March 12, 2021, and 202111460309.5 filed on December 2, 2021, all of which are hereby incorporated by reference in their entirety.

[0002] (Technical Field) The present invention relates to the field of medical technology, generally to heteroaryl compounds, and more particularly to novel aminoquinoline compounds that are inhibitors of the nucleotide - binding oligomerization domain (NOD) / receptor - interacting protein 2 (RIP2) pathway. Further, the present invention relates to compositions containing aminoquinoline compounds, methods for their preparation, and their applications in the treatment of RIP2 - related diseases including tumors, autoimmune diseases, neurodegenerative diseases, metabolic diseases, genetic diseases, etc., which are related to the prevention and / or treatment of diseases related to the inhibition of the receptor - interacting protein kinase 2 (RIP2).

Background Art

[0003] NOD - like receptors (NLRs) such as NOD1 and NOD2 (i.e., nucleotide - binding oligomerization domain - containing protein 1 and 2) are pattern - recognition receptors (PRRs). Activation of either NOD1 or NOD2 can lead to a cellular cascade reaction (Cell.Signal.18(2006)2223 - 2229).

[0004] The signal transduction mediated by NOD2 depends on receptor-interacting protein kinase 2 (RIP2). For example, usually, the leucine-rich repeat (LRR) domain of NOD2 is folded and enters the intermediate domain, thereby becoming self-inhibitory. However, when the LRR is recognized and bound by its substrate muramyl dipeptide (MDP), NOD2 changes its conformation and autoactivates (Science 300 (2003) 1584-1587; J. Biol. Chem. 278 (2003) 5509-5512), thereby recruiting and activating downstream RIP2 through the CARD-CRAD (i.e., caspase activation and recruitment domain) interaction between NOD2 and RIP2. Subsequently, RIP2 undergoes autophosphorylation and is ubiquitinated by a series of E3 ubiquitin ligases including X chromosome-linked inhibitor of apoptosis protein (XIAP) and cellular inhibitor of apoptosis protein 1 (cIAP1). Polyubiquitinated RIP2 oligomerizes, promoting the polyubiquitination of NF-κB essential modulator (NEMO, also known as inhibitor of nuclear factor kappa-B kinase subunit gamma (IKK-γ)) and the activation of transforming growth factor-β-activated kinase 1 (TAK1). Subsequently, it controls the recruitment of TAK1 adapter proteins TAB1 and TAB2 / 3, leading to the activation of the mitogen-activated protein kinase (MAPK) signaling pathway (extracellular signal-regulated kinase (ERK), p38, c-Jun N-terminal kinase (JNK)) and the activation of nuclear factor kappa B (NF-κB). For example, the TAB proteins of the TAK1 / TAB2 / TAB3 complex bind to the lysine-63 (K63)-linked polyubiquitin chain, thereby phosphorylating and activating the IKK complex. The activation of the IKK complex promotes the phosphorylation of the inhibitor of κB molecule α (IκBα), leading to the dissociation of IκB and the activation of the NF-κB pathway.The activated heterodimer p65 / p50 then translocates to the nucleus and activates the transcription of genes involved in immune response, cell death pathways, and growth control (Clinical Immunology (2021), 223, 108648).

[0005] Receptor-interacting protein kinase 2 (RIP2, also known as RIPk2, RICK, CARDIAK, or CARD3) is a member of the receptor-interacting serine / threonine protein kinase family involved in innate immune signaling and is encoded by the RIP2 gene on human chromosome 8. The protein encoded by 61 kDa has a C-terminal caspase recruitment domain (CARD), an N-terminal kinase domain, and a bridging intermediate domain (Curr. Biol. 8 (1998) 885-888).

[0006] RIP2 plays an important role in the immune system and is regulated by the intracellular peptidoglycan sensors NOD1 and NOD2 (J. Immunol. 178 (2007) 2380-2386), triggering the innate immune response against bacteria and infections. Initial studies suggested that the kinase activity of RIP2 was unnecessary for the activation of the NF-κB pathway and the production of cytokines. However, transgenic mice lacking the RIP2 protein were deficient in their response to NOD1 or NOD2 agonists, highlighting the important role of the RIP2 kinase site in NOD1 and NOD2 (J. Immunol. 2007, 178 (4), 2380-2386). When RIP2 is activated, serine 176 and tyrosine 474 are autophosphorylated. Phosphorylation of serine 176 is required for the activation of RIP2. However, phosphorylation of tyrosine 474 enhances the activity of RIP2 but is not required for signaling (Genes Dev. 24 (2010), 2666-2677).

[0007] Aberrant regulation of the NOD / RIP2-dependent signaling pathway is involved in a number of human diseases, including asthma, early-onset inflammatory bowel disease, sarcoidosis, Crohn's disease, multiple sclerosis, Blau syndrome (a hyper-rare autoinflammatory disease), etc. RIP2 is upregulated in conditions such as sepsis and Alzheimer's disease. Furthermore, RIP2 can function as a prognostic marker in different cancer types, such as inflammatory breast cancer (a rare and aggressive form of breast cancer with high mortality). RIP2 is overexpressed in patients with inflammatory breast cancer. References: EMBO Mol. Med. 5 (2013) 1278 - 1295; Arthritis Rheum. 43 (2013) 125 - 130; Pediatr. Rheumatol. Online J. 12 (2014) 33; Scientific World Journal 2016 (2016) 2597376; J. Leukoc. Biol. 94 (2013) 927 - 932; Biochem. Biophys. Res. Commun. 281 (2001) 84 - 93; Cancers (Basel) 10 (2018).

[0008] Inhibition of RIP2 kinase activity may inhibit downstream signals of NOD1 / 2. Therefore, by developing small molecule inhibitors that inhibit RIP2 kinase activity, the progression of disease conditions or pathologies caused by the activation of the NF-κB pathway or MAPK pathway can be delayed. As a result, a preventive effect or a therapeutic effect can be expected, and clinical applications are anticipated.

Summary of the Invention

[0009] The present disclosure provides quinoline derivatives as inhibitors of the NOD / RIP2 pathway, as well as their compositions and uses. These disclosed quinoline derivatives, as well as their compositions and uses, can effectively prevent or treat diseases and disorders responsive to RIP receptor inhibition, including, for example, tumors, autoimmune diseases, neurodegenerative diseases, metabolic diseases, and genetic diseases.

[0010] The objective of the present disclosure is to provide an RIP2 inhibitor, as well as its composition and use.

[0011] Aspects of the present disclosure relate to compounds of formula (I):

Chemical formula

Chemical formula

Chemical formula

[0012] In some embodiments of the aspects provided herein, R2 is H.

[0013] In some embodiments, L is a bond, O, or

Chemical formula

[0014] In some embodiments,

Chemical formula

Chemical formula

[0015] In some embodiments,

Chem.

Chem.

[0016] In some embodiments,

Chem.

Chem.

[0017] In some embodiments,

Chem.

Chem.

[0018] In some embodiments, R 3 is, independently, H, halide, -OH, amino, C 1-6 alkyl, C 3-6 cycloalkyl, -O(C 1-6 alkyl), 3-6 membered cycloheteroalkyl, 5-10 membered heteroaryl, where C 1-6 alkyl, C 3-6 cycloalkyl, 3-6 membered cycloheteroalkyl, 5-10 membered heteroaryl are unsubstituted or are substituted with 1-3 groups independently selected from R c herein.

[0019] In some embodiments, R 3is, independently, H, F, methyl, ethyl, n-propyl, i-propyl, methoxy, -OCD3, -OCF3, -OCHF2, -OCH2CH2OH, -OCH2CH2OCH3, [Chemical formula] where * indicates the connection to L.

[0020] In some embodiments, R 3 is, independently, H, methyl, ethyl, n-propyl, i-propyl, methoxy, -OCH2CH2OH, -OCH2CH2OCH3, [Chemical formula] where * indicates the connection to L.

[0021] In some embodiments, R 3 is, independently, H, F, methyl, methoxy, -OCD3, -OCF3, -OCHF2, -OCH2CH2OH, -OCH2CH2OCH3, [Chemical formula] where * indicates the connection to L.

[0022] In some embodiments, R 3 is, independently, F, methoxy, -OCD3, -OCF3, -OCHF2, -OCH2CH2OH, -OCH2CH2OCH3, [Chemical formula] where * indicates the connection to L.

[0023] In some embodiments, R 3 is, independently, methoxy, -OCD3, -OCF3, and -OCHF2, where * indicates the connection to L.

[0024] In some embodiments, R 3-L is independently H, F, methyl, ethyl, n-propyl, i-propyl, methoxy, -OCD3, -OCF3, -OCHF2, -OCH2CH2OH, -OCH2CH2OCH3,

Chem.

[0025] In some embodiments, R 4 is methyl, R 5 is methyl, and R 6 is H or C 1-3 alkyl. In some embodiments, R 4 is methyl; R 5 is methyl; and R 6 is H. In some embodiments, R 7 is H, deuterium, F, Cl, or Br. In some embodiments, R 7 is H or F.

[0026] In some embodiments, the compound of formula (I) is

Chem.

[0027] In some embodiments, the present disclosure provides a pharmaceutical composition comprising a therapeutically effective amount of a compound disclosed herein or a pharmaceutically acceptable salt, ester, solvate, prodrug, isotopically labeled derivative, stereoisomer or tautomer thereof, and a pharmaceutically acceptable carrier.

[0028] In some embodiments, the present disclosure provides a pharmaceutical formulation comprising a compound disclosed herein or a pharmaceutically acceptable salt, ester, solvate, prodrug, isotope-labeled derivative, stereoisomer or tautomer thereof, wherein the pharmaceutical formulation comprises tablets, capsules, injections, granules, powders, suppositories, pills, gels, dispersions, oral solutions, inhalation solutions, suspensions, or dry suspensions.

[0029] In some embodiments, the present disclosure provides a composition comprising: (i) a compound of the present disclosure, or a pharmaceutically acceptable salt, ester, solvate, prodrug, isotope-labeled derivative, stereoisomer or tautomer thereof, or a pharmaceutical composition of the present disclosure; and (ii) at least one additional therapeutic agent selected from the group consisting of anti-tumor agents, agents for treating autoimmune diseases, anti-neurodegenerative agents, agents for treating metabolic diseases, and agents for treating genetic diseases.

[0030] In some embodiments, the present disclosure provides a method for treating a disease or disorder associated with the RIP2 receptor in a mammal suffering therefrom, comprising administering to the mammal a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt, ester, solvate, prodrug, isotope-labeled derivative, or a pharmaceutical composition of the present disclosure, or a composition of the present disclosure, wherein the disease or disorder associated with the RIP2 receptor is a systemic inflammatory response, an autoimmune disease, a tumor, cancer, a metabolic disease or a neurodegenerative disease.

[0031] In some embodiments, the present disclosure provides a method for treating a disease or disorder associated with the RIP2 receptor in a mammal suffering therefrom, comprising administering to the mammal a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt, ester, solvate, prodrug, isotopically labeled derivative, stereoisomer or tautomer thereof, or a pharmaceutical composition of the present disclosure, or a composition of the present disclosure, wherein the disease or disorder associated with the RIP2 receptor is uveitis, dermatitis, acute pneumonia, type 2 diabetes, arthritis, ulcerative colitis, Crohn's disease, early-onset inflammatory bowel disease, extraintestinal inflammatory bowel disease, prevention of ischemia-reperfusion injury in solid organ transplantation, non-alcoholic steatohepatitis, autoimmune hepatitis, asthma, systemic lupus erythematosus, sarcoidosis, Wegener's granulomatosis, interstitial lung disease, pulmonary fibrosis, renal fibrosis, liver fibrosis, myocardial infarction, hypersensitivity pneumonia, ankylosing spondylitis, multiple sclerosis, systemic sclerosis, polymyositis, rheumatoid arthritis, myasthenia gravis, type 1 diabetes, glomerulonephritis, autoimmune thyroiditis, graft rejection, Crohn's disease, Blau syndrome, scleroderma, psoriasis, stomatitis, retinitis pigmentosa, proliferative vitreoretinopathy, Best vitelliform macular dystrophy, eczema, urticaria, vasculitis, eosinophilic fasciitis, wet age-related macular degeneration, dry age-related macular degeneration, diabetic retinopathy, retinopathy of prematurity (ROP), diabetic macular edema, uveitis, retinal vein occlusion, cystoid macular edema, glaucoma, Parkinson's disease, Alzheimer's disease, Huntington's disease, breast cancer, lung cancer, bladder cancer, pancreatic cancer, liver cancer, head and neck squamous cell carcinoma, thyroid cancer, sarcoma, osteosarcoma, desmoid tumor, melanoma, prostate cancer, colorectal cancer, ovarian cancer, cervical cancer, esophageal cancer, gastric cancer, myeloma, lymphoma, mantle cell lymphoma, cutaneous T cell lymphoma, chronic and non-progressive anemia, primary or essential thrombocythemia, leukemia, acute leukemia, chronic leukemia, lymphocytic leukemia, myeloid leukemia, myelodysplastic syndrome, myeloproliferative disorder, brain tumor, astrocytoma, medulloblastoma, schwannoma, primitive neuroectodermal tumor, or pituitary tumor.

[0032] In some embodiments, the compounds disclosed herein, or pharmaceutically acceptable salts, esters, solvates, prodrugs, isotope-labeled derivatives, stereoisomers or tautomers thereof, or the pharmaceutical compositions disclosed herein, are for use in methods of treating the human or animal body by therapy. In some embodiments, the compounds disclosed herein, or pharmaceutically acceptable salts, esters, solvates, prodrugs, isotope-labeled derivatives, stereoisomers or tautomers thereof, or the pharmaceutical compositions disclosed herein, are for use as pharmaceuticals. In some embodiments, the compounds disclosed herein, or pharmaceutically acceptable salts, esters, solvates, prodrugs, isotope-labeled derivatives, stereoisomers or tautomers thereof, or the pharmaceutical compositions disclosed herein, are for use in the treatment of diseases or disorders associated with the RIP2 receptor. Diseases or disorders associated with the RIP2 receptor are systemic inflammatory responses, autoimmune diseases, tumors, cancers, metabolic diseases, or neurodegenerative diseases.In some embodiments, the disease or disorder associated with the RIP2 receptor is uveitis, dermatitis, acute lung injury, type 2 diabetes, arthritis, ulcerative colitis, Crohn's disease, early-onset inflammatory bowel disease, extraintestinal inflammatory bowel disease, prevention of ischemia-reperfusion injury in solid organ transplantation, non-alcoholic steatohepatitis, autoimmune hepatitis, asthma, systemic lupus erythematosus, sarcoidosis, Wegener's granulomatosis, interstitial lung disease, pulmonary fibrosis, renal fibrosis, liver fibrosis, myocardial infarction, hypersensitivity pneumonia, ankylosing spondylitis, multiple sclerosis, systemic sclerosis, polymyositis, rheumatoid arthritis, myasthenia gravis, type 1 diabetes, glomerulonephritis, autoimmune thyroiditis, graft rejection, Crohn's disease, Blau syndrome, scleroderma, psoriasis, stomatitis, retinitis pigmentosa, proliferative vitreoretinopathy, Best vitelliform macular dystrophy, eczema, urticaria, vasculitis, eosinophilic fasciitis, wet age-related macular degeneration, age-related macular degeneration, diabetic retinopathy, retinopathy of prematurity (ROP), retinal macular edema, uveitis, retinal vein occlusion, cystoid macular edema, glaucoma, Parkinson's disease, Alzheimer's disease, Huntington's disease, breast cancer, lung cancer, bladder cancer, pancreatic cancer, liver cancer, head and neck squamous cell carcinoma, thyroid cancer, sarcoma, osteosarcoma, desmoid tumor, melanoma, prostate cancer, colorectal cancer, ovarian cancer, cervical cancer, esophageal cancer, gastric cancer, myeloma, lymphoma, mantle cell lymphoma, cutaneous T-cell lymphoma, chronic non-progressive anemia, primary or essential thrombocythemia, leukemia, acute leukemia, chronic leukemia, lymphocytic leukemia, myelogenous leukemia, myelodysplastic syndrome, myeloproliferative disorder, brain tumor, astrocytoma, medulloblastoma, schwannoma, primitive neuroectodermal tumor, or pituitary tumor.

[0033] In some embodiments,

[0034] In some embodiments, the compound of formula (II):

Chemical formula

Chem.

Chem.

[0035] In some embodiments, the compound of formula (III):

Chem.

Chem.

Chemical formula

[0036] The present disclosure also provides a compound disclosed herein, a pharmaceutical composition disclosed herein, or a formulation of a composition disclosed herein, wherein the formulation is a tablet, capsule, injection, granule, powder, suppository, pill, gel, powder, oral solution, inhalant, suspension, or dry suspension.

[0037] Additional aspects and advantages of the present disclosure will become readily apparent to those skilled in the art from the following detailed description, which illustrates and describes only exemplary embodiments of the present disclosure. As will be realized, the present disclosure is capable of other and different embodiments, and some of the details thereof are capable of modification in various obvious respects all without departing from the present disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature and not restrictive.

Brief Description of the Drawings

[0038]

Figure 1

[0039]

Figure 2

[0040]

Figure 3

[0041]

Figure 4

[0042]

Figure 5

[0043] Before proceeding with the detailed description, it should be understood that the following detailed description is merely exemplary in nature and is not intended to limit the invention or its application and use. Accordingly, the present disclosure has been depicted and described as shown in specific exemplary embodiments for the sake of convenience of explanation, but it will be understood that it can be implemented in various other types of embodiments and equivalents, as well as in various other systems and environments. Furthermore, there is no intention to be bound by any theory presented in the preceding background or the following detailed description.

[0044] (Incorporation by reference) All publications, patents, and patent applications mentioned herein are hereby incorporated by reference to the same extent as if each individual publication, patent, or patent application were specifically and individually indicated to be incorporated by reference. [Embodiments for Carrying Out the Invention]

[0045] Although various embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Without departing from the present invention, numerous variations, modifications, and substitutions may occur to those skilled in the art. It should be understood that various alternatives to the embodiments of the present invention described herein may be employed.

[0046] (Definition) Compounds are generally described herein using standard nomenclature. It should be understood that for compounds having an asymmetric center, all optical isomers and mixtures thereof are included (unless otherwise specified). Further, compounds having a carbon-carbon double bond may exist in Z- and E-forms, and all isomeric forms of such compounds are included in the present invention unless otherwise specified. When a compound exists in various tautomers, the described compound is not limited to any one particular tautomer, but rather is intended to include all tautomers.

[0047] As used herein, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a molecule" includes a plurality of such molecules, and the like.

[0048] The term "about" or "substantially" as used herein generally means within ±15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of a specified amount.

[0049] As used herein, the terms "halogen" or "halide" generally mean fluorine, chlorine, bromine, and iodine. As used herein, the term "haloalkyl" generally refers to an alkyl group substituted with one or more independently selected halogens (e.g., a "C1-C6 haloalkyl" group has 1-6 carbon atoms and at least one halogen). Examples of haloalkyl groups include, but are not limited to, mono-, di- or trifluoromethyl; mono-, di- or trichloromethyl; mono-, di-, tri-, tetra- or pentafluoroethyl; mono-, di-, tri-, tetra- or pentachloroethyl; and 1,2,2-tetrafluoro-L-trifluoromethyl-ethyl, etc.

[0050] As used herein, the term "alkyl" generally means a straight or branched chain saturated aliphatic hydrocarbon. Alkyl groups include, for example, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, n-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, and 3,3-dimethyl-2-butyl, and include groups having 1-8 carbon atoms (C 1-8 alkyl), 1-6 carbon atoms (C 1-6 alkyl) and 1-4 carbon atoms (C1-C4 alkyl). Similarly, C 1-3 alkyl refers to an alkyl group having 1-3 straight or branched carbon atoms, and examples include methyl, ethyl, propyl and isopropyl. In some cases, the substituents of the alkyl group are specifically indicated. For example, "cyanoalkyl" refers to an alkyl group substituted with at least one cyano substituent. In some embodiments, C 1-6 alkyl is preferably methyl, ethyl, n-propyl, isopropyl, or tert-butyl.

[0051] As used herein, the term "alkenyl" generally means a straight-chain or branched-chain alkene group containing at least one unsaturated carbon-carbon double bond. The alkenyl group is C 2-8 alkenyl, C 2-6 alkenyl and C 2-4 including alkenyl groups, each having 2-8, 2-6 or 2-4 carbon atoms, such as, for example, ethenyl, allyl or isopropenyl. As used herein, the term "alkynyl" generally refers to a straight-chain or branched-chain alkyne group, which has one or more unsaturated carbon-carbon bonds, at least one of which is a triple bond. The alkynyl group includes C 2-8 alkynyl, C 2-6 alkynyl and C 2-4 alkynyl groups, each having 2-8, 2-6 or 2-4 carbon atoms.

[0052] As used herein, the term "alkoxy" generally refers to an alkyl group as described above bonded to another chemical moiety via an oxygen bridge. The alkoxy group includes, for example, C 1-6 alkoxy and C 1-4 alkyl groups of different lengths such as alkoxy groups, each having 1-6 or 1-4 carbon atoms. As used herein, the term "OC 1-6 alkyl" generally means that the alkoxy group contains an alkyl group (having 1-6 carbon atoms) bonded to an oxygen atom. Methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, tert-butoxy, n-pentoxy, 2-pentoxy, 3-pentoxy, isopentoxy, neopentoxy, hexy, 2-hexy, 3-hexy and 3-methylpentoxy are representative alkoxy groups.

[0053] As used herein, the term "cycloalkyl" generally means a group consisting of one or more saturated rings in which all ring members are carbon. For example, a particular cycloalkyl group is C 3-8Cycloalkyl, in which case the cycloalkyl group contains one or more rings having from 3 to 8 ring members, all of which are carbon, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Other examples of cycloalkyl groups include adamantyl and the like. The cycloalkyl group does not form an aromatic ring or a heterocyclic ring. As used herein, the term "cycloalkenyl" generally means a group consisting of one or more unsaturated rings in which all ring members are carbon.

[0054] As used herein, the terms "heterocyclic ring", "heterocycle", or "cycloheteroalkyl" generally refer to a ring structure (monocyclic or polycyclic) containing 3 - 12 ring atoms (3 - 12 membered heterocyclic ring), 3 - 8 ring atoms (3 - 8 membered heterocyclic ring or 3 - 8 membered cycloheteroalkyl), 3 - 6 ring atoms (3 - 6 membered heterocyclic ring or 3 - 6 membered cycloheteroalkyl), or 5 - 6 ring atoms (5 - 6 membered heterocyclic ring or 5 - 6 membered cycloheteroalkyl), wherein at least one ring atom is carbon and at least one ring atom is a heteroatom selected from N, O, and S, or a heteroatom group is selected from C(=O), S(=O), S(=O)2. The heterocyclic group may be aromatic or non - aromatic. Piperidine and oxetane are non - limiting examples of non - aromatic heterocyclic rings. Thiazole and pyridine are non - limiting examples of aromatic heterocyclic rings. Other examples of heterocyclic rings include aziridinyl, azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, piperidinyl, morpholinyl, piperazinyl, thiomorpholinyl, tetrahydropyranyl, 1,1 - dioxothiomorpholinyl, butyrolactam, valerolactam, caprolactam, butyrolactone, valerolactone, caprolactone. Similarly, the term "cycloheteroalkenyl" refers to a monocyclic or polycyclic structure composed of carbon atoms (s) and heteroatom(s) / heteroatom group(s), and cycloheteroalkenyl consists of at least one C = C double bond, at least one ring atom that is carbon, and at least one ring atom that is a heteroatom selected from N, O, and S, or a heteroatom group selected from C(=O), S(=O), S(=O)2.

[0055] "Aryl" refers to a monocyclic or fused - ring polycyclic group having 6 - 12 carbon atoms with a fully conjugated π - electron system (C 6-12 aryl) or 6 - 10 aryl (C 6-10It refers to those with all carbon atoms of (aryl). Examples of aryl groups include, but are not limited to, phenyl, naphthalenyl, tetrahydronaphthyl, indanyl, biphenyl, and anthracenyl. The aryl group may be substituted or unsubstituted. Representative substituents include halo, trihalomethyl, alkyl, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, nitro, carbonyl, thiocarbonyl, C-carboxy, O-carboxy, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amide, N-amide, sulfinyl, sulfonyl, amino and -NRXRY, where RX and RY are independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, carbonyl, acetyl, sulfonyl, trifluoromethanesulfonyl, and a 5- or 6-membered heterocyclic alicyclic ring in combination. Exemplary substituted alkyl groups include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, aminomethyl, aminoethyl, hydroxymethyl, methoxymethyl, 2-fluoroethyl, and 2-methoxyethyl, etc.

[0056] As used herein, the term "heteroaryl" generally refers to an aromatic group in which at least one aromatic ring contains at least one heteroatom selected from N, O, and S, and heteroaryl is, for example, a 5- to 12-membered heteroalkyl, a 5- to 10-membered heteroalkyl, a 5- to 7-membered monocyclic structure, or a 7- to 12-membered bicyclic structure. The number of heteroatoms in heteroaryl can be 1, 2, 3, 4, or more. For example, thienyl, pyridyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridin-2(1H)-one, pyridin-4(1H)-one, pyrrolyl, pyrazolyl, thiazolyl, 1,2-,3-triazolyl, 1,2,4-triazolyl, 1,2,-5-oxadiazolyl, imidazolyl, furanyl, tetrazolyl, isothiazolyl, oxazolyl, thiadiazolyl, oxadiazolyl, naphthyl, benzothienyl, indolyl, benzimidazolyl, benzothiazolyl, benzofuranyl, quinolinyl, isoquinolinyl, and quinazolinyl, and is not limited thereto. The heteroaryl group may be substituted or unsubstituted. Typical substituents include halo, trihalomethyl, alkyl, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, nitro, carbonyl, thiocarbonyl, C-carboxy, O-carboxy, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amide, N-amide, sulfinyl, sulfonyl, amino, and -NRXRY, where RX and RY are as defined above.

[0057] As used herein, the term "amino" generally refers to a primary amino group (-NH2), a secondary amino group (-NH-), and a tertiary amino group (

Chemical formula

[0058] As used herein, the term "alkylamino" generally refers to a secondary or tertiary amine having the general structure of NH-R 1 or N(R 1 )(R 2 ), where R1 and R 2 is independently selected from alkyl, cycloalkyl and (cycloalkyl)alkyl groups. Such groups include, for example, mono- and di-(C 1-6 alkyl)amino groups, but are not limited thereto, and each C 1-6 alkyl may be the same or different. It will be apparent that the definition of "alkyl" used in the term "alkylamino" differs from the definition of "alkyl" used for all other alkyl-containing groups in that it includes cycloalkyl groups and (cycloalkyl)alkyl groups.

[0059] As used herein, the term "alkylthio" generally refers to an alkyl-substituted thio group, where the term alkyl is as defined above.

[0060] As used herein, the terms "substituent" and "substituted" generally refer to a molecular moiety being covalently bonded to an atom within the molecule of interest. For example, a ring substituent may be a moiety such as a halogen, an alkyl group, a haloalkyl group, etc., that is covalently bonded to an atom (preferably a carbon atom or a nitrogen atom) that is a ring member. Substituents of an aromatic group are generally covalently bonded to a ring carbon atom. A straight-chain substituent may be a site such as a halogen, an alkyl group, a haloalkyl group, etc., that is covalently bonded to an atom (preferably a carbon atom or a nitrogen atom) that is a straight-chain member.

[0061] As used herein, the term "bicycloheteroalkyl" generally means a bicyclic structure that shares one or two atoms and contains at least one heteroatom independently selected from the group consisting of N, O, and S in the ring. As used herein, the term "bicycloheteroalkylene" generally means a diradical of a bicycloheteroalkyl group that may be bonded to two other groups.

[0062] As used herein, the term "cycloalkylamine" generally means either a ring structure in which an amino group is bonded to a carbon atom within the ring or a ring structure having a nitrogen atom as a member of the ring.

[0063] As used herein, the term "cycloalkylamide" generally refers to either a ring structure having an amide group bonded to a carbon atom in the ring via an amide carbon or a ring structure in which both the amide nitrogen and the amide carbon atom are members of the ring.

[0064] As used herein, the term "cyclourea" generally means a ring structure in which the urea carbon and both urea nitrogen atoms are members of the ring. An example of a cyclourea is oxoimidazolidine.

[0065] As used herein, the term "pharmaceutically acceptable" generally means a form of a compound that is safe for administration to a subject. For example, free base, salt form, solvate, hydrate, prodrug or derivative forms of the compounds described herein that are approved by a regulatory authority such as the US Food and Drug Administration (FDA) for mammalian use by oral ingestion or other routes of administration are pharmaceutically acceptable.

[0066] Compounds of formula (I), (II) or (III) are in the form of pharmaceutically acceptable salts of the free base compounds. As used herein, the term "pharmaceutically acceptable salt" generally refers to salts that are commonly used to form alkali metal salts and addition salts of free acids or free bases and are approved by regulatory authorities. Salts are formed from ionic bonds, charge-charge interactions, covalent bonds, complex formation, coordination, etc. The nature of the salt is not important as long as it is pharmaceutically acceptable.

[0067] As used herein, the term "pharmaceutically acceptable salt" means a salt that, within the scope of sound medical judgment, is suitable for use in contact with the tissues of a subject without undue toxicity, irritation, allergic response, etc., and that has a reasonable benefit / risk ratio. For example, Berge et al., in Pharmaceutical Sciences (1977) 66:1-19, describe pharmaceutically acceptable salts in detail. Pharmaceutically acceptable salts of the compounds provided herein include those derived from suitable inorganic and organic acids and bases. Examples of inorganic acids from which salts can be derived include, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. Examples of organic acids from which salts can be derived include, but are not limited to, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc. Examples of pharmaceutically acceptable non-toxic acid addition salts include salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, perchloric acid, etc., or organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, malonic acid, etc., or by other methods used in the art such as ion exchange.Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, besylate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. In some embodiments, organic acids from which the salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.

[0068] Pharmaceutically acceptable salts derived from suitable bases include alkali metals, alkaline earth metals, ammonium, and other amine salts. Inorganic bases from which the salts can be derived include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Organic bases from which the salts can be derived include primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, and are not limited thereto. Examples include, but are not limited to, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salts are ammonium salts, potassium salts, sodium salts, calcium salts, or magnesium salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Further pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium, and amine cations formed with counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkylsulfonates, arylsulfonates, and the like. Organic bases from which the salts can be derived include, for example, primary amines, secondary amines, tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, and examples include those such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salts are selected from ammonium salts, potassium salts, sodium salts, calcium salts, and magnesium salts. Bis salts (i.e., two counterions) and higher salts (e.g., three or more counterions) are included within the meaning of pharmaceutically acceptable salts.

[0069] As used herein, the term "ester" refers to an organic compound containing an ester linkage including monoester, diester, triester, and polyester.

[0070] As used herein, the term "solvate" refers to a compound further comprising a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces. The solvate can be the disclosed compound or a pharmaceutically acceptable salt thereof. When the solvent is water, the solvate is a "hydrate". Other solvates include, but are not limited to, methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran, dimethyl sulfoxide, and N,N-dimethylformamide. Pharmaceutically acceptable solvates and hydrates can be complexes that contain, for example, from 1 to about 100, or from 1 to about 10, or from 1 to about 2, 3, or 4, solvent or water molecules.

[0071] As used herein, and unless otherwise specified, "prodrug" means a compound that can be converted to a biologically active compound described herein under physiological conditions or by solvolysis. Thus, the term "prodrug" means a precursor of a pharmaceutically acceptable biologically active compound. A prodrug can be inactive when administered to a subject but is converted in vivo to an active compound, for example, by hydrolysis. Discussions of prodrugs are described in Higuchi, T., et al., "Pro-drugs as Novel Delivery Systems," A.C.S. Symposium Series, Vol. 14, and Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference in their entirety. The term "prodrug" also means any covalent carrier that, when such prodrug is administered to a mammalian subject, releases the active formula (I), (II), or (III) in vivo. Prodrugs of the active compounds described herein can be prepared by modifying the functional groups present in the active formula (I), (II) or (III) such that the modification is cleaved either by routine manipulation or in vivo to give the parent active compound. Prodrugs include compounds in which a hydroxy group, an amino group or a mercapto group is bonded to any group that cleaves to give a free hydroxy group, a free amino group or a free mercapto group, respectively, when the prodrug of the active formula (I), (II) or (III) is administered to a mammalian subject.

[0072] The terms "isotopically labeled", "isotopically labeled", "isotopically labeled derivative" and "isotopically labeled" refer to an unnatural proportion of atomic isotopes in one or more of the atoms that make up such a compound. For example, a compound can be radioactively labeled with a radioactive isotope such as, for example, tritium ( 3 H), iodine 125 ( 125 I), carbon 14 ( 14 C), etc. Also, a compound can be 2 H, 11 C, 13 C, 15 N, 17 O, 18 O, 18 F, 32 P, 35 S, 36 Cl. Certain isotopically labeled disclosed compounds (e.g., those labeled with 3 H and 14 C) are useful in tissue distribution assays of the compound and / or substrate. Tritium (i.e., 3 H) and carbon 14 (i.e., 14C) Isotopes can enable ease of preparation and detectability. Further, by substituting with heavy isotopes such as deuterium (i.e., 2H), certain therapeutic advantages resulting from higher metabolic stability (e.g., increased in vivo half-life or decreased dosage) can be obtained. Isotope-labeled disclosed compounds can generally be prepared by replacing an isotope-labeled reagent with an unlabeled reagent. In some embodiments, provided herein are compounds that can also include unnatural proportions of atomic isotopes in one or more of the atoms that make up such compounds. All isotope variations of the compounds of the present disclosure, whether radioactive or not, are encompassed within the scope of the present disclosure.

[0073] As used herein, the term “isomer” generally refers to different compounds having the same molecular formula, including all geometric and stereoisomers. “Stereoisomers” are isomers that differ only in the way the atoms are arranged in space. For example, “isomers” include geometric double bond cis- and trans-isomers, also known as E- and Z-isomers; R- and S-enantiomers; diastereomers, (d)-isomers and (l)-isomers, their racemic mixtures; and other mixtures thereof, as falling within the scope of this disclosure unless otherwise specified. As used herein, the term “tautomer” is a type of isomer that includes two or more interconvertible compounds resulting from at least one formal shift of a hydrogen atom and at least one change in valence (e.g., from a single bond to a double bond, from a triple bond to a single bond, or vice versa).

[0074] In some embodiments, the compound(s) of formula (I), (II) or (III) are used to treat a subject by administering them as a pharmaceutical composition. For this purpose, the compound(s) are, in one embodiment, combined with one or more pharmaceutically acceptable excipients including a carrier, diluent or adjuvant to form a suitable composition, which is described in more detail herein.

[0075] As used herein, the term "excipient" generally refers to any pharmaceutically acceptable additive, carrier, adjuvant, or other suitable component other than the pharmaceutically active ingredient (API) typically included for the purposes of formulation and / or administration.

[0076] As used herein, the term "diluent" generally means an agent used as a filler to achieve the desired volume or weight of a composition. A diluent can be present in a pharmaceutical composition within granules in the form of a single compound or in the form of a mixture of compounds. Non-limiting examples of diluents include lactose, starch, pregelatinized starch, microcrystalline cellulose, siliconized microcrystalline cellulose, cellulose acetate, dextrose, mannitol, sodium phosphate, potassium phosphate, calcium phosphate, fructose, maltose, sorbitol, or sucrose.

[0077] As used herein, the term "adjuvant" generally refers to any substance or mixture of substances that increases the potency or efficacy of a compound disclosed herein against a target when the adjuvant is used in conjunction with the compound disclosed herein. However, when the adjuvant is used alone, no pharmacological effect is observed against the same target.

[0078] As used herein, the terms "treating," "treat," "treatment," and "therapy" generally, but not limited to, refer to therapies including therapeutic treatment, prophylactic treatment, and preventative treatment. Prophylactic treatment generally constitutes either completely preventing the onset of a disorder or delaying the onset of a pre-clinical manifest stage of a disorder in an individual. Treatment includes the medical management of a patient aimed at curing, improving, stabilizing, or preventing a disease, pathological condition, or disorder. This term includes active treatment, i.e., treatment specifically aimed at improving a disease, pathological condition, or disorder, and causal treatment, i.e., treatment aimed at removing the cause of a related disease, pathological condition, or disorder. Further, this term includes palliative treatment, i.e., treatment aimed at alleviating symptoms rather than curing a disease, pathological condition, or disorder, prophylactic treatment, i.e., treatment aimed at minimizing or partially or completely suppressing the onset of a related disease, pathological condition, or disorder, and supportive treatment, i.e., treatment used to supplement another specific treatment aimed at improving a related disease, pathological condition, or disorder.

[0079] As used herein, the terms "prevent" or "prevention" refer to preventing, avoiding, interfering with, forestalling, stopping, or hindering something from happening, particularly by prior action. In this specification, when reduction, inhibition, or prevention is used, it is understood that the use of the other two words is also explicitly disclosed, unless otherwise indicated.

[0080] As used herein, the vocabulary "effective amount" generally refers to the quantification of the amount of each agent that, while avoiding side effects typically associated with alternative therapies, by itself achieves the goal of improving the severity and frequency of occurrence of a disorder over the treatment with each agent. The effective amount is, in one embodiment, administered in a single dosage form or in multiple dosage forms.

[0081] Regardless of the selected route of administration, the compounds of the present invention, and / or the pharmaceutical compositions of the present invention, which may be used in the appropriate hydrated form, are formulated by pharmaceutically acceptable dosage forms or by other conventional methods known to those skilled in the art.

[0082] The actual dosage of the active ingredient in the pharmaceutical composition of the present invention can be varied to obtain an effective amount of the active ingredient that is non-toxic to the patient and achieves the desired therapeutic response for a particular patient, composition, and mode of administration.

[0083] The dosage selected will be determined by a variety of factors including the activity of the particular compound of the invention employed, the route of administration, the time of administration, the rate of excretion of the particular compound employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular hedgehog inhibitor employed, the age, sex, weight, condition, general health and prior history of the patient being treated, and like factors well known in the medical arts.

[0084] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the required pharmaceutical composition. For example, the physician or veterinarian can initiate the dosage of the compound of the invention employed in the pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0085] Generally, a preferred daily dosage of the compounds of the present invention will be that amount of the compound that is the lowest dosage effective to produce a therapeutic effect. Such effective amounts generally depend upon the above factors. Generally, the intravenous, intracerebral and subcutaneous dosages of the compounds of the present invention for a patient will range from about 0.0001 to about 100 mg per kg of body weight per day. The mode of administration can have a significant effect on the dosage. In the case of local routes of administration, higher dosages can be used.

[0086] Optionally, if desired, the effective daily amount of the active compound can be administered in unit dosage form as 2, 3, 4, 5, 6 or more sub-dosages administered separately at appropriate intervals throughout the day. One of ordinary skill in the art will readily understand that the dosage levels can vary as a function of the particular compound, the severity of the symptoms, and the susceptibility of the subject to side effects. The dosages of the particular compounds disclosed herein can be readily determined by one of ordinary skill in the art by a variety of means.

[0087] (Pharmaceutical Composition and Preparation) One embodiment provides a pharmaceutical composition comprising a compound of formula (I), (II) or (III), or a stereoisomer, tautomer, hydrate, solvate or pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0088] In some embodiments, the compounds described herein are formulated into pharmaceutical compositions. The pharmaceutical compositions are formulated by conventional methods using one or more pharmaceutically acceptable inert ingredients that facilitate processing the active compound into a pharmaceutically usable formulation. Appropriate formulation depends on the chosen route of administration. An overview of pharmaceutical compositions described herein is, for example, described in Remington: The Science and Practice of Pharmacy, 19th Edition, Easton, Pa.: Mack Publishing Company (1995); Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania (1975); Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y. (1980); and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Edition, Lippincott Williams & Wilkins (1999), which disclosures are hereby incorporated by reference herein.

[0089] As used herein, a pharmaceutical composition refers to a mixture of a compound of formula (I), (II) or (III) and other chemical components (i.e., pharmaceutically acceptable inert ingredients) such as, for example, carriers, excipients, binders, fillers, suspending agents, flavoring agents, sweetening agents, disintegrants, dispersing agents, surfactants, lubricants, coloring agents, diluents, solubilizing agents, wetting agents, plasticizers, stabilizers, penetration enhancers, wetting agents, defoaming agents, antioxidants, preservatives, or one or more combinations thereof. The pharmaceutical composition facilitates the administration of the compound to a living being. In the practice of the therapeutic or use methods provided herein, a therapeutically effective amount of the compounds described herein is administered in a pharmaceutical composition to a mammal having a disease, disorder, or condition to be treated. In some embodiments, the mammal is a human. The therapeutically effective amount can vary widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors. The compounds can be used alone or in combination with one or more therapeutic agents as components of a mixture.

[0090] The pharmaceutical formulations described herein are administered to a subject by suitable routes of administration including, but not limited to, oral, parenteral (e.g., intravenous, subcutaneous, intramuscular), intranasal, buccal, topical, rectal, or transdermal routes. The pharmaceutical formulations described herein include, but are not limited to, aqueous liquid dispersions, self-emulsifying dispersions, solid solutions, liposomal dispersions, aerosols, solid dosage forms, powders, immediate release formulations, controlled release formulations, fast dissolve formulations, tablets, capsules, pills, delayed release formulations, sustained release formulations, pulsatile release formulations, multiparticulate formulations, and mixed immediate and controlled release formulations.

[0091] All formulations for oral administration are dosage units suitable for such administration. Examples of such dosage units include tablets or capsules. In some embodiments, these contain an amount of active ingredient of from about 1 to 2000 mg, preferably from about 1 to 500 mg, typically from about 5 to 150 mg. The daily dosage suitable for humans or other mammals varies widely depending on the condition of the patient and other factors, but can again be determined using routine methods and practice.

[0092] Conventional compounding techniques include, for example, one or a combination of the methods of (1) dry mixing, (2) direct compression, (3) grinding, (4) dry or non-aqueous granulation, (5) wet granulation, or (6) melting. Other methods include, for example, spray drying, pan coating, melt granulation, granulation, fluidized bed spray drying or coating (e.g., Wurster coating), tangential coating, top spray, tableting, extrusion, and the like.

[0093] (Synthesis method) The method of the present invention may involve the use of at least one compound of formula (I), (II) or (III) that inhibits programmed necrosis in the repair and / or modulation of functional performance of a wide range of cells, tissues and organs, and may have therapeutic and cosmetic uses ranging from the regulation of the formation and repair of nervous tissue, bone and cartilage, the regulation of spermatogenesis, the regulation of smooth muscle, the regulation of the lungs, liver and other organs arising from the primitive gut, the regulation of hematopoietic function, the regulation of skin and hair growth, etc. Accordingly, the methods and compositions of the present invention include the use of inhibitors for all such uses in which an inhibitor of programmed necrosis may be involved. Further, the subject method can be carried out on cells provided in culture (in vitro) or on cells of an entire animal (in vivo).

[0094] The examples and preparations provided below illustrate and exemplify the compounds described herein and methods for preparing such compounds. Generally, the compounds described herein can be prepared by processes known in general chemical techniques.

[0095] The compounds of the present invention can be prepared using various synthetic routes including those described below, starting from commercially available materials. The starting materials of the present invention are either known, commercially available, or can be synthesized analogously to, or in accordance with, methods known in the art. Many starting materials can be prepared according to known processes, in particular, using the processes described in the examples. When synthesizing the starting materials, functional groups are optionally protected with appropriate protecting groups where necessary. The functional groups can be removed according to known procedures in the art.

[0096] The protection of functional groups by protecting groups, the protecting groups themselves, and their removal reactions (generally referred to as "deprotection") are described, for example, in standard references such as J.F.W. McOmie, Protective Groups in Organic Chemistry, Plenum Press, London and New York (1973), T.W. Greene, Protective Groups in Organic Synthesis, Wiley, New York (1981), The Peptides, Volume 3, E. Gross and J. Meienhofer editors, Academic Press, London and New York (1981).

[0097] All synthetic procedures described herein can be carried out under known reaction conditions, preferably under the conditions described herein, either in the absence or presence (usually) of a solvent or diluent.

[0098] The present invention further encompasses "intermediate" compounds, whether isolated or not, containing structures generated from the described synthetic procedures prior to finally obtaining the desired compound. Structures resulting from performing steps from transient starting materials, structures resulting from deviating from the described method(s) at any stage, and structures forming starting materials under reaction conditions are all "intermediates" included in the present invention. Furthermore, structures resulting from using starting materials in the form of reactive derivatives or salts, or structures resulting from compounds obtained by the means of the processes according to the present invention, and structures resulting from treating the compounds of the present invention in situ are also within the scope of the present invention.

[0099] New starting materials and / or intermediates, and processes for their preparation are likewise the subject of the present invention. In selected embodiments, such starting materials are used to obtain the desired compound(s), and the reaction conditions are selected.

[0100] The starting materials of the present invention are those that can be known, commercially available, or synthesized analogously to, or in accordance with, methods known in the art. Many starting materials can be prepared according to known processes, in particular, using the processes described in the examples. When synthesizing starting materials, functional groups are, in some cases, protected with appropriate protecting groups if necessary. Protecting groups, their introduction and removal are as described above.

[0101] All reagents and solvents were obtained commercially unless otherwise specified. All commercially available reagents and solvents were used without purification unless otherwise specified. When necessary, some reagents and solvents were purified by standard techniques. For example, tetrahydrofuran can be purified by distillation from sodium. All thin-layer chromatography (TLC, GF254) analyses and column purifications (100 - 200 mesh) were performed on silica gel (Qingdao Haiyang Chemical Co., Ltd. or Yantai Chemical Co., Ltd.), using petroleum ether (BPM 60 - 90 °C) / ethyl acetate (v / v) as the eluent; and spots were visualized by UV at 254 nm and I2 vapor, or phosphomolybdic acid. All organic layers after extraction were dried over anhydrous Na2SO4 unless otherwise stated. All nuclear magnetic resonance spectra ( 1 1H NMR) were recorded on a Varian-400 spectrometer at 400 MHz using TMS as the internal standard. LC-MS was performed using an Agilent 1100 system equipped with an LC-MSD Trap recorder, a diode array detector (DAD) with detection wavelengths of 214 nm and 254 nm, and an ESI source. The HPCL column was an Agela Durashell C18 3.5 μm 4.6×50 mm column. The gradient was run using 0.1 N aqueous NH4HCO3 and acetonitrile, with a gradient from 5 / 95 to 95 / 5 over the indicated runtime (e.g., 5 minutes) at a flow rate of 1.8 mL / min.

[0102] The size and scale of the synthetic method vary depending on the desired amount of the final product. Specific reactants and amounts are provided in the examples, but it is understood that those skilled in the art know other alternative and equally viable sets of reactants that can yield the same compound. Thus, when common oxidizing agents, reducing agents, and solvents of various properties (non-polar, apolar, polar, etc.) are utilized, equivalents are known in the art and are intended to be used in this method.

[0103] Many of the following steps describe various workups after the completion of the reaction. Workup generally involves quenching the reaction to terminate any remaining catalytic activity and starting reagents. This is typically followed by the addition of an organic solvent and separation of the aqueous layer from the organic layer. The product is usually obtained from the organic layer, while unreacted starting materials, other false by-products, and unwanted chemicals are generally trapped in the aqueous layer and discarded. Workup in standard organic synthesis procedures found in the literature generally involves drying the product by exposure to a drying agent such as anhydrous Na2SO4 to remove excess water or aqueous by-products that are partially dissolved in the organic layer, followed by concentration of the remaining organic layer. Concentration of the product dissolved in the solvent can be achieved by any known means, such as evaporation under pressure, evaporation under elevated temperature and pressure, etc. Such concentration can be achieved by using standard laboratory equipment such as a rotary evaporator distillation. Subsequently, optionally, one or more purification steps may be performed, including, but not limited to, flash column chromatography, filtration through various media, and / or other preparative methods known in the art, and / or crystallization / recrystallization. (See, for example, Addison Ault, “Techniques and Experiments for Organic Chemistry,” 6th ed., University Science Books, Sausalito, Calif., 1998, Ann B. McGuire, Ed., pp. 45-59).

[0104] (Abbreviations)

[0105] DCM means dichloromethane. DCE means 1,2-dichloroethane. DMF means N,N-dimethylformamide. EtOAc or EA means ethyl acetate. MeOH means methyl alcohol. EtOH means ethyl alcohol. Ph2O means diphenyl ether. Dioxane is 1,4-dioxane. Xantphos is (9,9-dimethyl-9H-xanthene-4,5-diyl)bis(diphenylphosphane). Pd2(dba)3 is tris(dibenzylideneacetone)dipalladium(0). DEAD is diethyl azodicarboxylate. NBS is N-bromosuccinimide. CDI is 1,1'-carbonyldiimidazole. THF is tetrahydrofuran. PMNH2 is 4-methoxybenzylamine. Et3N is triethylamine. Con.HCl or conc.HCl means concentrated hydrochloric acid. Sol.HCl means dilute hydrochloric acid. TLC means thin layer chromatography. HATU means 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate. DIPEA means diisopropylethylamine. HPLC means high performance liquid chromatography. LC-MS means liquid chromatography - mass spectrometer. NMR means nuclear magnetic resonance.

[0106] (General synthetic route) The following methods A - J are embodiments of some general synthetic routes leading to the compounds of formula (I), (II) or (III). The detailed reaction conditions for each method can be confirmed in the examples shown below.

[0107] Method A: [Chemical formula]

[0108] When benzothiazol-5-amine was fluorinated with Selectfluor, the corresponding product was obtained (step a).

[0109] Method B

Chemical formula

[0110] After 5 - amino - 2 - bromophenol was acetylated with acetic anhydride and then subjected to the Mitsunobu reaction, N-(4 - bromo - 3 - ((tetrahydrofuran - 3 - yl)oxy)phenyl)acetamide (steps a, b) was obtained. When deacetylated with concentrated hydrochloric acid, the corresponding compound was obtained (step c).

[0111] Method C

Chemical formula

[0112] Commercially available aniline was reacted with 5-(methoxymethylene)-2,2 - dimethyl - 1,3 - dioxane - 4,6 - dione and cyclized at high temperature in an inert solvent to produce a quinoline derivative (steps a, b). Chlorination with POCl3 and coupling with dimethylphosphine oxide gave the corresponding intermediate (steps c, d). The final compound was obtained by the S N Ar reaction with each aromatic amine (step e).

[0113] Method D

Chemical formula

[0114] Demethylation of the 7 - methoxy group of 6 - bromo - 4 - chloro - 7 - methoxyquinoline using BBr3, followed by alkylation of the newly exposed hydroxyl group with an alkyl bromide to obtain the corresponding intermediate (steps a, b). S NThe Ar reaction was carried out in the presence of concentrated hydrochloric acid (step c). The resulting compound was coupled with dimethylphosphine oxide to obtain the final product (step d).

[0115] Method E

Chem.

[0116] After coupling with dimethylphosphine oxide using a palladium catalyst, the corresponding final compound was obtained by the Suzuki coupling reaction with each arylboronic acid (steps a, b).

[0117] Method F

Chem.

[0118] Deprotection of the 1,3-dioxolane-containing intermediate with HCl.EA gave the final compound (step a).)

[0119] Method G

Chem.

[0120] Bromination of commercially available 2-amino-4-methoxybenzoic acid with NBS gave a brominated derivative (step a). This was reacted with nitromethane under basic conditions to generate the corresponding intermediate (step b). Intramolecular cyclization using CDI generated quinoline (step c). The nitro group was reduced with iron powder to generate an amine, which was converted to fluorine by the Sandmeyer reaction (steps e, f). Condensation with benzothiazol-5-amine and a coupling reaction gave the final compound (steps g, h).

[0121] Method H

Chem.

[0122] The oxetane-containing intermediate was obtained by nickel-catalyzed coupling of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline and 3-iodooxetane (step a). Treatment with NBS gave 4-bromo-3-(oxetan-3-yl)aniline, which was reacted with 5-(methoxymethylene)-2,2-dimethyl-1,3-dioxane-4,6-dione (steps b, c). Intramolecular cyclization reaction produced quinoline (step d). Activation of the hydroxyl group with Tf2Of and backward Hartwig coupling with benzothiazol-5-amine gave the corresponding intermediate (step e). The final compound was obtained by coupling reaction with dimethylphosphine oxide (step f).

[0123] Method I

Chemical formula

[0124] 1-(5-Bromo-2-chloropyridin-3-yl)ethan-1-one was obtained by condensation of 5-bromo-2-chloronicotinic acid and N,O-dimethylhydroxylamine, followed by Grignard reaction with methylmagnesium bromide. After treatment with DMF-DMA and then with PMB-NH2, 6-bromo-1-(4-methoxybenzyl)-1,8-naphthyridin-4(1H)-one was obtained. Deprotection of the PMB group gave 6-bromo-1,8-naphthyridin-4-ol, and its hydroxyl group was converted to chlorine with POCl3. S N Ar reaction was carried out under acidic conditions, followed by a coupling reaction to obtain the final compound.

[0125] Method J

Chemical formula

[0126] After brominating 2-amino-4-methoxybenzonitrile with NBS and then performing a Grignard reaction with methylmagnesium bromide, 1-(2-amino-5-bromo-4-methoxyphenyl)ethan-1-one was obtained. Treatment with NaNO2 gave a quinoline derivative. The hydroxyl group was converted to chlorine with POCl3 and reacted with benzo[d]thiazol-5-amine and S N Ar to give the corresponding intermediate. The target product was obtained by coupling with methylphosphine oxide using a palladium catalyst.

[0127] Example 1. Method A

[0128] Synthesis of 4-fluorobenzo[d]thiazol-5-amine

Chemical formula

[0129] Step a. 4-fluorobenzo[d]thiazol-5-amine: To a solution of benzo[d]thiazol-5-amine (3.0 g, 20 mmol) in acetonitrile (80 mL) was added selected fluorine (7.0 g, 20 mmol) in acetonitrile (20 mL). The mixture was stirred at room temperature for 1 hour. Water (200 mL) was added to the reaction solution, and the organic layer was extracted with dichloromethane (100 mL × 3). The combined organic layers were dried over Na2SO4 and concentrated under vacuum. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 4 / 1) to give the desired product (310 mg, 9.2%) as a yellow solid. 1 1H NMR (300 MHz, CDCl3) δ 8.92 (s, 1H), 7.48 (d, J = 8.4 Hz, 1H), 6.97 (t, J = 8.1 Hz, 1H), 3.87 (s, 2H). LC-MS (m / z): 169.0 [M + H] + .

[0130] Example 2. Method B

[0131] Synthesis of 4-bromo-3-((tetrahydrofuran-3-yl)oxy)aniline [Chem.]

[0132] Step a. N-(4-Bromo-3-hydroxyphenyl)acetamide: 5-Amino-2-bromophenol (660 mg, 3.5 mmol) was dissolved in acetic acid (5 mL), and acetic anhydride (396 mg, 3.9 mmol) was added. The mixture was stirred at room temperature for 10 minutes. Water (200 mL) was added, and the resulting solid was filtered and dried under vacuum to obtain the desired product (600 mg, 74%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 10.21 (s, 1H), 9.93 (s, 1H), 7.47 (s, 1H), 7.33 (d, J = 8.8 Hz, 1H), 6.86 (d, J = 8.4 Hz, 1H), 2.01 (s, 3H). LC-MS (m / z): 229.8 [M+H] + .

[0133] Step b. N-(4-Bromo-3-((tetrahydrofuran-3-yl)oxy)phenyl)acetamide: To a solution of N-(4-bromo-3-hydroxyphenyl)acetamide (600 mg, 2.61 mmol) in dry tetrahydrofuran (5 mL) were added PPh3 (1.37 g, 5.22 mmol) and diethyl azodicarboxylate (909 mm, 5.22 mmol). The mixture was stirred under nitrogen at room temperature for 30 minutes. Tetrahydrofuran-3-ol (275 mg, 3.13 mmol) was added, and the mixture was stirred for 2 hours. The solvent was removed in vacuo, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 5 / 1) to obtain the crude product (960 mg) as a white solid. LC-MS (m / z): 299.7 [M+H] + .

[0134] Step c. 4-Bromo-3-((tetrahydrofuran-3-yl)oxy)aniline: Concentrated HCl (4 mL) was added to a solution of the crude intermediate N-(4-bromo-3-((tetrahydrofuran-3-yl)oxy)phenyl)acetamide (960 mg) in ethanol (15 mL). The mixture was stirred at 85 °C for 3 hours. The solvent was concentrated to give the corresponding crude product (620 mg). LC-MS (m / z): 257.8 [M+H]+.

[0135] Example 3. Method C

[0136] (4-(Benzo[d]thiazol-5-ylamino)quinolin-6-yl)dimethylphosphine oxide hydrochloride (A1) was synthesized. [Chemical formula]

[0137] Step a. 5-(((4-Iodophenyl)amino)methylene)-2,2-dimethyl-1,3-dioxane-4,6-dione: 5-(Methoxymethylene)-2,2-dimethyl-1,3-dioxane-4,6-dione (1.3 g, 6.85 mmol) was added to a solution of 4-iodoaniline (1.0 g, 4.57 mmol) in methanol (15 mL). The mixture was stirred at room temperature for 30 minutes. The resulting solid was collected by filtration, washed with ethanol (3 mL), and dried under vacuum to give the desired product (1.2 g, 70%) as a pale yellow solid. 1 1H NMR (400 MHz, DMSO-d6) δ 11.21 (d, J = 14.4 Hz, 1H), 8.54 (d, J = 14.4 Hz, 1H), 7.76 (d, J = 8.0 Hz, 2H), 7.40 (d, J = 8.0 Hz, 2H), 1.67 (s, 6H).

[0138] Step b. 6-Iodoquinolin-4-ol: Diphenyl ether (140 mL) was added to a round-bottom flask, and the solvent was heated to 240 °C for 20 minutes. Intermediate 5-(((4-iodophenyl)amino)methylene)-2,2-dimethyl-1,3-dioxane-4,6-dione (1.2 g, 3.2 mmol) was slowly added. The mixture was stirred for 3 minutes. After cooling to room temperature, petroleum ether (80 mL) was added to the reaction mixture, and the resulting solid was collected by filtration, washed with ethyl acetate (20 mL), and dried under vacuum to give the target product (850 mg, 98%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 11.87 (s, 1H), 8.36 (s, 1H), 7.97 - 7.87 (m, 2H), 7.37 (d, J = 8.4 Hz, 1H), 6.07 (d, J = 7.2 Hz, 1H). LC-MS (m / z): 271.7 [M+H] + .

[0139] Step c. 4-Chloro-6-iodoquinoline: 6-Iodoquinolin-4-ol (200 mg, 0.74 mmol) was added to POCl3 (5 mL), and the mixture was stirred at reflux for 15 minutes to obtain a light brown solution. After cooling to room temperature, the excess POCl3 was removed in vacuo. The residue was dissolved in ethyl acetate (10 mL). The pH was adjusted to 7 using saturated NaHCO3 solution. The organic layer was extracted with ethyl acetate (60 mL × 2). The combined organic layers were dried over Na2SO4 and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 5 / 1) to give the target product (200 mg, 94%). 1 H NMR (400 MHz, DMSO-d6) δ 8.87 (d, J = 4.0 Hz, 1H), 8.54 (s, 1H), 8.15 (d, J = 8.8 Hz, 1H), 7.88 (d, J = 8.8 Hz, 1H), 7.81 (d, J = 4.0 Hz, 1H).

[0140] Step d. (4-Chloroquinolin-6-yl)dimethylphosphine oxide: To a solution of 4-chloro-6-iodoquinoline (200 mg, 0.69 mmol) in 1,4-dioxane (5 mL) was added a solution of dimethylphosphine oxide (81 mg, 1.04 mmol), Et3N (118 mg, 1.17 mmol), Pd2dba3 (32 mg, 0.03 mmol) and Xantphos (40 mg, 0.07 mmol). The mixture was stirred overnight at room temperature under a N2 atmosphere. The solvent was removed in vacuo and the residue was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 50 / 1) to afford the desired product (100 mg, 60%). 1 1H NMR (400 MHz, DMSO-d6) δ 8.96 (d, J = 4.4 Hz, 1H), 8.64 (d, J = 12.4 Hz, 1H), 8.27 - 8.13 (m, 2H), 7.89 (d, J = 4.4 Hz, 1H), 1.79 (s, 3H), 1.76 (s, 3H).

[0141] Step e. (4-(Benzo[d]thiazol-5-ylamino)quinolin-6-yl)dimethylphosphine oxide hydrochloride: (4-Chloroquinolin-6-yl)dimethylphosphine oxide (100 mg, 0.42 mmol) was dissolved in ethanol (3 mL), followed by the addition of benzo[d]thiazol-5-amine (67 mg, 0.46 mmol). The mixture was stirred at reflux for 1 h. After cooling to room temperature, the resulting solid was collected by filtration, washed with ethanol and dried in vacuo to give the desired product (50 mg, 34%) as the hydrochloride salt.

[0142] Example 4. Method D

[0143] (4-(Benzo[d]thiazol-5-ylamino)-7-(2-hydroxyethoxy)quinolin-6-yl)dimethylphosphine oxide (A7) was synthesized.

Chemical Structure

[0144] Step a. 6-Bromo-4-chloroquinolin-7-ol: To a solution of 6-bromo-4-chloro-7-methoxyquinoline (2.8 g, 10.3 mmol) in 1,2-dichloroethane (10 mL) was added BBr3 (10.3 mL, 31.0 mmol) at room temperature. The mixture was stirred at 110 °C for 1 h under microwave irradiation. The reaction was quenched with saturated aqueous Na2SO3 and extracted with dichloromethane (50 mL × 2). The combined organic layers were dried over Na2SO4 and concentrated. The residue was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 50 / 1) to give the desired product (1.7 g, 64%). 1 1H NMR (400 MHz, DMSO-d6) δ 11.86 (s, 1H), 8.87 (d, J = 5.2 Hz, 1H), 8.42 (s, 1H), 7.75 (d, J = 5.2 Hz, 1H), 7.55 (s, 1H). LC-MS (m / z): 257.7 [M + H] + .

[0145] Step b. 2-((6-Bromo-4-chloroquinolin-7-yl)oxy)ethan-1-ol: To a solution of 6-bromo-4-chloroquinolin-7-ol (200 mg, 0.77 mmol) in N,N-dimethylformamide (3 mL) was added K2CO3 (215 mg, 1.55 mmol). The mixture was stirred at 80 °C for 30 min. Subsequently, 2-bromoethanol (193 mg, 1.55 mmol) was added and the mixture was stirred at 80 °C overnight. The solvent was removed in vacuo and the residue was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 50 / 1) to give the desired product (150 mg, 64%). LC-MS (m / z): 301.7 [M + H] + .

[0146] Step c. 2-((4-(Benzo[d]thiazol-5-ylamino)-6-bromoquinolin-7-yl)oxy)ethan-1-ol hydrochloride: To a solution of 2-((6-bromo-4-chloroquinolin-7-yl)oxy)ethan-1-ol (150 mg, 0.5 mmol), benzo[d]thiazol-5-amine (93 mg, 0.62 mmol) was added. The mixture was stirred at reflux for 30 minutes. After cooling to room temperature, the resulting solid was collected by filtration, washed with ethanol, and dried in vacuo to obtain the desired product (120 mg, 58%) as the hydrochloride salt. 1 H NMR (400 MHz, DMSO-d6) δ 14.27 (s, 1H), 11.03 (s, 1H), 9.53 (s, 1H), 9.17 (s, 1H), 8.45 (d, J = 6.4 Hz, 1H), 8.37 (d, J = 8.4 Hz, 1H), 8.20 (s, 1H), 7.60 (d, J = 8.4 Hz, 1H), 7.56 (s, 1H), 6.81 (d, J = 6.8 Hz, 1H), 4.28 (s, 2H), 3.88 (s, 2H). LC-MS (m / z): 415.6 [M+H] + .

[0147] Step d. (4-(Benzo[d]thiazol-5-ylamino)-7-(2-hydroxyethoxy)quinolin-6-yl)dimethylphosphine oxide: To the solution, 2-((4-(benzo[d]thiazol-5-ylamino)-6-bromoquinolin-7-yl)oxy)ethan-1-ol hydrochloride (120 mg, 0.29 mmol) was dissolved in 1,4-dioxane (3 mL), and dimethylphosphine oxide (34 mg, 0.43 mmol), Et3N (50 mg, 0.49 mmol), Pd2dba3 (26 mg, 0.03 mmol), and Xantphos (18 mg, 0.03 mmol) were added. The mixture was stirred at 125 °C for 1 hour under microwave irradiation. The solvent was removed in vacuo, and the residue was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 20 / 1) to obtain the target product (30 mg, 25%).

[0148] Example 5. Method E

[0149] Synthesis of (4-(Benzo[d]thiazol-5-ylamino)-7-(1-methyl-1H-pyrazol-4-yl)quinolin-6-yl)dimethylphosphine oxide (A11) [Chemical formula]

[0150] Step a. 2-Bromo-4-nitroaniline: To a solution of 4-nitroaniline (10 g, 72.5 mmol) in dichloromethane (60 mL) was added NBS (13 g, 72.5 mmol). The mixture was stirred at room temperature overnight. Saturated aqueous NaHCO3 was added to quench the reaction. The organic layer was separated, dried over Na2SO4, and concentrated to give the crude product (15 g, 96%) as a yellow solid. LC-MS (m / z): 216.7 [M+H] + .

[0151] Step b. 2-Bromo-1-iodo-4-nitrobenzene: To a solution of 2-bromo-4-nitroaniline (7.0 g, 32.3 mmol) in acetic acid (80 mL) was added a solution of NaNO2 (2.4 g, 34.8 mmol) in conc. H2SO4 (16 mL) at 0 °C. The reaction mixture was stirred for 4 hours. A mixture of KI (16 g, 96.9 mmol) and I2 (3.5 g, 32.3 mmol) dissolved in water (60 mL) was added, and the reaction mixture was stirred at room temperature overnight. 15% aqueous NaOH was added to quench the reaction. The organic layer was extracted with ethyl acetate (300 mL), dried over Na2SO4, and concentrated to give the desired product (10.0 g, 95%).

[0152] Step c. 3-Bromo-4-iodoaniline: To a solution of 2-bromo-1-iodo-4-nitrobenzene (10.0 g, 30.7 mmol) in ethanol (60 mL) were added NH4Cl (8.2 g, 153 mmol) and iron powder (8.5 g, 153 mmol). The mixture was stirred at 85 °C for 2 hours. The reaction mixture was filtered through diatomaceous earth, and the cake was washed with ethanol. The resulting filtrate was concentrated, and the residue was purified by silica gel column chromatography to give the desired product (6.0 g, 65%) as a yellow solid. 11H NMR (400 MHz, CDCl3) δ 8.45 (s, 1H), 8.08 (d, J = 8.8 Hz, 1H), 7.85 (d, J = 8.4 Hz, 1H), LC-MS (m / z): 297.6 [M+H] + .

[0153] Step d. 7-Bromo-6-iodoquinolin-4-ol: To a solution of 3-bromo-4-iodoaniline (6.0 g, 20 mmol) in ethanol (30 mL) was added 5-(methoxymethylene)-2,2-dimethyl-1,3-dioxane-4,6-dione (7.4 g, 40 mmol). The mixture was stirred at room temperature for 2 h. The resulting solid was collected by filtration and dried in vacuo to give the crude product. Ph2O (30 mL) was added to a round-bottom flask, followed by the addition of the crude product. The reaction was stirred at 220 °C for 3 min. After cooling to room temperature, the resulting solid was filtered, washed with ethyl ether, and dried in vacuo to give the desired product (4.0 g). LC-MS (m / z): 349.5 [M+H] + .

[0154] Step e. N-(7-Bromo-6-iodoquinolin-4-yl)benzo[d]thiazol-5-amine: 7-Bromo-6-iodoquinolin-4-ol (2.0 g, 5.75 mmol) was added to POCl3 (20 mL). The mixture was stirred at 110 °C for 2 h. The solvent was removed in vacuo. The residue was dissolved in ethyl acetate (20 mL × 2) and subsequently concentrated. The resulting solid was dissolved in i-PrOH (15 mL), and benzo[d]thiazol-5-amine (950 mg, 6.33 mol) was added. The mixture was stirred at 95 °C overnight. The solvent was removed in vacuo, and the pH was adjusted to 8 using saturated aqueous NaHCO3. The organic layer was extracted with ethyl acetate (100 mL) and concentrated. The residue was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 20 / 1) to give the desired product (950 mg, 35%) as a yellow solid. 11H NMR (400 MHz, DMSO-d6) δ 9.44 (s, 1H), 9.09 (s, 1H), 8.48 (d, J = 5.6 Hz, 1H), 8.24 - 8.21 (m, 2H), 8.04 (d, J = 2.0 Hz, 1H), 7.53 (dd, J = 4.2, 2.0 Hz, 1H), 7.01 (d, J = 5.6 Hz, 1H). LC-MS (m / z): 481.4 [M+H] + .

[0155] Step f. (4-(Benzo[d]thiazol-5-ylamino)-7-bromoquinolin-6-yl)dimethylphosphine oxide: To a solution of N-(7-bromo-6-iodoquinolin-4-yl)benzo[d]thiazol-5-amine (800 mg, 1.60 mmol) in 1,4-dioxane (10 mL) were added Pd2dba3 (73 mg, 0.08 mmol), Xantphos (93 mg, 0.16 mmol), dimethylphosphine oxide (187 mg, 2.40 mmol) and Et3N (323 mg, 3.20 mmol). The mixture was stirred at 70 °C overnight under a N2 atmosphere. The solvent was removed in vacuo and the resulting residue was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 50 / 1) to give the corresponding product (240 mg, 33%) as a yellow solid. 1 1H NMR (400 MHz, DMSO-d6) δ 11.52 (s, 1H), 9.53 (s, 1H), 9.18 (d, J = 12.8 Hz, 1H), 8.55 (d, J = 6.8 Hz, 1H), 8.42 - 8.31 (m, 2H), 8.19 (s, 1H), 7.59 (d, J = 8.4 Hz, 1H), 6.92 (d, J = 6.4 Hz, 1H), 2.00 (s, 3H), 1.96 (s, 3H). LC-MS (m / z): 432.0 [M+H] + .

[0156] Step g. (4-(Benzo[d]thiazol-5-ylamino)-7-(1-methyl-1H-pyrazol-4-yl)quinolin-6-yl)dimethylphosphine oxide: To a solution of (4-(benzo[d]thiazol-5-ylamino)-7-bromoquinolin-6-yl)dimethylphosphine oxide in 1,4-dioxane / H2O (10.0 mL / 0.5 mL) were added Pd(dppf)Cl2 (10 mg, 0.014 mmol), (1-methyl-1H-pyrazol-4-yl)boronic acid (35 mg, 0.28 mmol) and K2CO3 (40 mg, 0.28 mmol). The mixture was stirred at 100 °C overnight under a N2 atmosphere. The solvent was removed in vacuo and the resulting residue was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 10 / 1) to afford the desired product (10 mg, 17%) as a green solid.

[0157] Example 6. Method F

[0158] (S)-(4-(Benzo[d]thiazol-5-ylamino)-7-(2,3-dihydroxypropoxy)quinolin-6-yl)dimethylphosphine oxide (A18) synthesis [Chemical formula]

[0159] Step a. (S)-(4-(Benzo[d]thiazol-5-ylamino)-7-(2,3-dihydroxypropoxy)quinolin-6-yl)dimethylphosphine oxide: To a solution of (R)-(4-(benzo[d]thiazol-5-ylamino)-7-((2,2-dimethyl-1,3-dioxolan-4-yl)methoxy)quinolin-6-yl)dimethylphosphine oxide (120 mg, 0.25 mmol) in ethanol (5 mL) was added 3N hydrochloric acid in ethyl acetate (2 mL). The mixture was stirred at room temperature for 2 h. The solvent was removed and the residue was dissolved in dichloromethane / methanol (20 mL / 10 mL). The solvent was concentrated and the residue was purified by 18C column chromatography (water / methanol (v / v) = 60 / 40) to afford the title compound (90 mg, 82%) as a yellow solid.

[0160] Example 7. Method G

[0161] Synthesis of (4-(benzo[d]thiazol-5-ylamino)-3-fluoro-7-methoxyquinolin-6-yl)dimethylphosphine oxide (A23)

Chemical formula

[0162] Step a. 2-Amino-5-bromo-4-methoxybenzoic acid: To a solution of 2-amino-4-methoxybenzoic acid (15.0 g, 89.7 mmol) in N,N-dimethylformamide (50 mL) was slowly added NBS (16.0 g, 89.7 mmol) at 0 °C. The mixture was stirred at room temperature for 1 hour. The solvent was concentrated and the resulting solid was washed with ethyl acetate / petroleum ether (200 mL / 400 mL) to obtain the target product as a gray solid (18.0 g, 82%). 1 1H NMR (400 MHz, DMSO-d6) δ 7.76 (s, 1H), 6.41 (s, 1H), 3.79 (s, 3H). LC-MS (m / z): 246.0 [M+H] + .

[0163] Step b. 5-Bromo-4-methoxy-2-((2-nitrovinyl)amino)benzoic acid: To a solution of NaOH (53.9 g, 1347.5 mmol) in water (50 mL) was added nitromethane (21.9 g, 359.2 mmol). The mixture was stirred at 45 °C for 5 minutes. Additional nitromethane (21.9 g, 359.2 mmol) was added at room temperature. The mixture was stirred at room temperature for 10 minutes and at 50 °C for 5 minutes. The solvent was poured onto ice (500 g) and the pH was adjusted to ~2 with concentrated hydrochloric acid. This solvent was added to water (300 mL) containing 2-amino-5-bromo-4-methoxybenzoic acid (22.0 g, 89.8 mmol). Concentrated hydrochloric acid (161.5 mL) was added to the solution and the mixture was stirred overnight. The resulting solid was collected by filtration and dried in vacuo to obtain the desired product (24.0 g, 85%) as a yellow solid. 11H NMR (300 MHz, DMSO-d6) δ 13.07 (d, J = 13.5 Hz, 1H), 8.19 (dd, J = 13.2, 6.3 Hz, 1H), 8.09 (s, 1H), 7.35 (s, 1H), 6.82 (d, J = 6.3 Hz, 1H), 3.99 (s, 3H). LC-MS (m / z): 314.9 [M-H] - .

[0164] Step c. 6-Bromo-7-methoxy-3-nitroquinolin-4(1H)-one: CDI (1.5 g, 9.45 mmol) was added to a solution of 5-bromo-4-methoxy-2-((2-nitrovinyl)amino)benzoic acid (2.0 g, 6.3 mmol) in N,N-dimethylformamide (300 mL). The mixture was stirred at 60 °C overnight. The solvent was removed in vacuo and the resulting solid was washed with acetonitrile (300 mL) to afford the desired product (1.4 g, 72%) as a brown solid. 1 1H NMR (300 MHz, DMSO-d6) δ 8.58 (s, 1H), 7.76 (s, 1H), 6.67 (s, 1H), 6.64 (s, 1H), 3.42 (s, 3H). LC-MS (m / z): 299.0 [M+H] + .

[0165] Step d. 6-Bromo-4-chloro-7-methoxy-3-nitroquinoline: N,N-Dimethylformamide (2 mL) was added to a solution of 6-bromo-7-methoxy-3-nitroquinolin-4(1H)-one (14.0 g, 46.9 mmol) in POCl3 (100 mL). The mixture was stirred at 110 °C overnight. The POCl3 was removed in vacuo. The residue was dissolved in dichloromethane / water (100 mL / 100 mL) and stirred for 30 minutes. The organic layer was separated and the aqueous phase was extracted with dichloromethane (200 mL × 2). The combined organic layers were dried over Na2SO4 and purified by silica gel column chromatography (dichloromethane) to afford the desired product (3.36 g, 23%) as a white solid. 1 1H NMR (300 MHz, CDCl3) δ 9.25 (s, 1H), 8.64 (s, 1H), 7.51 (s, 1H), 4.11 (s, 3H).

[0166] Step e. 6-Bromo-4-chloro-7-methoxyquinolin-3-amine: To a solution of 6-bromo-4-chloro-7-methoxy-3-nitroquinoline (3.4 g, 10.6 mmol) in i-PrOH / H2O (100 mL / 25 mL) was added iron powder (3.0 g, 53.2 mmol) and NH4Cl (2.8 g, 53.2 mmol). The mixture was stirred at 75 °C for 2 h. The solvent was filtered through diatomaceous earth. The filtrate was extracted with dichloromethane (100 mL × 3). The combined organic layers were dried over Na2SO4 and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 2 / 1) to give the desired product as a brown solid (2.5 g, 86%). 1 H NMR (300 MHz, DMSO-d6) δ 8.54 (s, 1H), 8.04 (s, 1H), 7.41 (s, 1H), 5.96 (s, 2H), 3.94 (s, 3H). LC-MS (m / z): 287.0 [M+H] + .

[0167] Step f. 6-Bromo-4-chloro-3-fluoro-7-methoxyquinoline: To a solution of 6-bromo-4-chloro-7-methoxyquinolin-3-amine (2.5 g, 8.74 mmol) in dry tetrahydrofuran (20 mL) was added nitrosonium tetrafluoroborate (1.1 g, 9.42 mmol) at -10 °C. The mixture was stirred at 0 °C for 50 min. The resulting solid was filtered and dissolved in decahydronaphthalene. The mixture was stirred at 170 °C for 5 min. The solvent was evaporated and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 4 / 1) to give the target product as a white solid (435 mg, 17%). 1 H NMR (300 MHz, CDCl3) δ 8.76 (s, 1H), 8.39 (s, 1H), 7.46 (s, 1H), 4.05 (s, 3H). LC-MS (m / z): 289.9 [M+H] + .

[0168] Step g. N-(6-Bromo-3-fluoro-7-methoxyquinolin-4-yl)benzo[d]thiazol-5-amine: To a solution of 6-bromo-4-chloro-3-fluoro-7-methoxyquinoline (435 mg, 1.5 mmol) in ethanol (30 mL) was added benzo[d]thiazol-5-amine (225 mg, 1.5 mmol) and a drop of concentrated hydrochloric acid. The mixture was stirred at 85 °C for 2 h. The solvent was removed in vacuo and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 2 / 1) to give the title compound (85 mg, 14%) as a white solid. 1 H NMR (300 MHz, DMSO-d6) δ 9.35 (s, 1H), 9.24 (s, 1H), 8.77 (d, J = 3.3 Hz, 1H), 8.63 (s, 1H), 8.04 (d, J = 8.7 Hz, 1H), 7.59 (s, 1H), 7.50 (s, 1H), 7.23 (d, J = 7.5 Hz, 1H), 4.01 (s, 3H). LC-MS (m / z): 403.9 [M+H] + .

[0169] Step h. (4-(Benzo[d]thiazol-5-ylamino)-3-fluoro-7-methoxyquinolin-6-yl)dimethylphosphine oxide: To a solution of N-(6-bromo-3-fluoro-7-methoxyquinolin-4-yl)benzo[d]thiazol-5-amine (90 mg, 0.22 mmol) in 1,4-dioxane (3 mL) were added dimethylphosphine oxide (19 mg, 0.24 mmol), Pd2(dba)3 (20 mg, 0.02 mmol), Xantphos (13 mg, 0.02 mmol) and Et3N (45 mg, 0.12 mmol). The mixture was stirred at 125 °C for 2 h under a N2 atmosphere through microwave irradiation. The reaction was concentrated and the resulting solid was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 20 / 1) to give the crude product. This solid was washed with ethyl acetate / ether (1 mL / 4 mL) to give the title compound (14 mg, 16%) as a yellow solid.

[0170] Example 8. Method H

[0171] Synthesis of 4-(Benzo[d]thiazol-5-ylamino)-7-(oxetan-3-yl)quinolin-6-yl)dimethylphosphine oxide (A26)

Chem.

[0172] Step a. 3-(Oxetan-3-yl)aniline: 3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (4.0 g, 18.3 mmol), NiI2 (562 mg, 1.8 mmol) and trans-2-aminocyclohexanol hydrochloride (274 mg, 1.8 mmol) were added to a microwave tube, followed by addition of NaHMDs (9.15 mL, 18.3 mmol) and dry i-PrOH (20 mL). The mixture was stirred at room temperature for 10 minutes under a N2 atmosphere. 3-Iodooxetane (3.4 g, 18.3 mmol) was added and the mixture was stirred at 120 °C for 2 hours under a N2 atmosphere with microwave irradiation. The reaction was quenched with water and the organic layer was extracted with dichloromethane (50 mL × 2). The combined organic layers were dried over Na2SO4 and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 4 / 1) to give the desired product (1.01 g, 37%) as a yellow oil. 1 H NMR (300 MHz, CDCl3) δ 7.21 - 7.09 (m, 1H), 6.81 - 6.71 (m, 2H), 6.60 (d, J = 8.7 Hz, 1H), 5.04 (dd, J = 8.4, 6.0 Hz, 2H), 4.76 (t, J = 6.3 Hz, 2H), 4.21 - 4.06 (m, 1H), 3.70 (s, 2H). LC-MS (m / z): 150.1 [M + H] + .

[0173] Step b. 4-Bromo-3-(oxetan-3-yl)aniline: To a solution of 3-(oxetan-3-yl)aniline (1.01 g, 6.7 mmol) in acetonitrile (15 mL) was added NBS (961 mg, 5.4 mmol) in acetonitrile (5 mL) at 0 °C. The mixture was stirred at this temperature for 30 minutes. The solvent was concentrated and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 4 / 1) to give the title compound (913 mg, 60%) as a white solid. 1 H NMR (300 MHz, CDCl3) δ 7.31 - 7.24 (m, 1H), 6.78 (d, J = 2.4 Hz, 1H), 6.47 (dd, J = 8.7, 2.7 Hz, 1H), 5.05 (dd, J = 7.8, 6.0 Hz, 2H), 4.77 (t, J = 6.6 Hz, 2H), 4.59 - 4.45 (m, 1H), 3.77 (s, 2H). LC-MS (m / z): 228.0 [M+H] + .

[0174] Step c. 5-(((4-Bromo-3-(oxetan-3-yl)phenyl)amino)methylene)-2,2-dimethyl-1,3-dioxane-4,6-dione: To a solution of 4-bromo-3-(oxetan-3-yl)aniline (913 mg, 4.0 mmol) in ethanol (8 mL) was added 5-(methoxymethylene)-2,2-dimethyl-1,3-dioxane-4,6-dione (893 mg, 4.8 mmol). The mixture was stirred at room temperature for 30 minutes. The resulting solid was filtered, washed with ethanol and dried in vacuo to give the title compound (1.18 g, 77%) as a yellow solid. 1 H NMR (300 MHz, CDCl3) δ 11.28 (d, J = 14.4 Hz, 1H), 8.63 (d, J = 14.0 Hz, 1H), 7.61 (d, J = 8.3 Hz, 1H), 7.31 - 7.27 (m, 1H), 7.12 - 7.02 (m, 1H), 5.17 - 5.07 (m, 2H), 4.84 - 4.74 (m, 2H), 4.67 - 4.52 (m, 1H), 1.60 (s, 6H).

[0175] Step d. 6-Bromo-7-(oxetan-3-yl)quinolin-4-ol diphenyl ether (30 mL) was added to a round-bottom flask, and the solvent was heated to 240 °C for 20 minutes. Intermediate 5-(((4-bromo-3-(oxetan-3-yl)phenyl)amino)methylene)-2,2-dimethyl-1,3-dioxane-4,6-dione (1.18 g, 3.1 mmol) was slowly added to the solution. The mixture was stirred for 2 minutes. After cooling to room temperature, the resulting solid was filtered, washed with ether and dried in vacuo to give the target product (490 mg, 56%) as a yellow solid. 1 H NMR (300 MHz, DMSO-d6) δ 11.80 (s, 1H), 8.19 (s, 1H), 7.96 (d, J = 6.9 Hz, 1H), 7.64 (s, 1H), 6.07 (d, J = 7.5 Hz, 1H), 5.11 - 4.93 (m, 2H), 4.68 (t, J = 6.3 Hz, 2H), 4.63 - 4.47 (m, 1H). LC-MS (m / z): 280.0 [M+H] + .

[0176] Step e. N-(6-Bromo-7-(oxetan-3-yl)quinolin-4-yl)benzo[d]thiazol-5-amine: To a solution of 6-bromo-7-(oxetan-3-yl)quinolin-4-ol (250 mg, 0.89 mmol) in dichloromethane (4 mL) were added pyridine (703 mg, 8.9 mmol) and (CF3SO2)2O (1.25 g, 4.45 mmol) dropwise at 0 °C. The mixture was stirred at room temperature for 30 minutes. The solvent was concentrated, and the residue was dissolved in dry 1,4-dioxane (4 mL). Benzo[d]thiazol-5-amine (161 mg, 1.07 mmol), Pd2dba3 (82 mg, 0.09 mmol), Xantphos (52 mg, 0.09 mmol) and Cs2CO3 (870 mg, 2.67 mmol) were added to the solution. The mixture was stirred at 100 °C for 10 minutes. The solvent was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 20 / 1) to give the target product as a gray solid (130 mg, 35%). 11H NMR (300 MHz, DMSO-d6) δ 9.43 (s, 1H), 9.30 (s, 1H), 8.77 (s, 1H), 8.51 (s, 1H), 8.20 (d, J = 8.1 Hz, 1H), 8.02 (s, 1H), 7.94 (s, 1H), 7.53 (d, J = 7.5 Hz, 1H), 7.02 (s, 1H), 5.11 - 4.96 (m, 2H), 4.90 - 4.73 (m, 2H), 4.71 - 4.53 (m, 1H). LC-MS (m / z): 412.0 [M+H] + .

[0177] Step f. (4-(Benzo[d]thiazol-5-ylamino)-7-(oxetan-3-yl)quinolin-6-yl)dimethylphosphine oxide: To a solution of N-(6-bromo-7-(oxetan-3-yl)quinolin-4-yl)benzo[d]thiazol-5-amine (120 mg, 0.29 mmol) in 1,4-dioxane (4 mL) was added dimethylphosphine oxide (45 mg, 0.58 mmol), Cs2CO3 (284 mg, 0.87 mmol), Pd2dba3 (27 mg, 0.03 mmol), and Xantphos (17 mg, 0.03 mmol). The mixture was stirred at 130 °C for 2.5 h under microwave irradiation. The solvent was removed in vacuo, and the residue was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 100 / 9) to afford the title compound (200 mg, 17%) as a yellow solid.

[0178] Example 9. Method I

[0179] (5-(Benzo[d]thiazol-5-ylamino)-1,8-naphthyridin-3-yl)dimethylphosphine oxide (B1) synthesis

Chem.

[0180] Step a. 5-Bromo-2-chloro-N-methoxy-N-methylnicotinamide: To a solution of 5-bromo-2-chloronicotinic acid (3.0 g, 12.7 mmol) in N,N-dimethylformamide (30 mL) was added CDI (3.1 g, 19.1 mmol). The mixture was stirred at room temperature for 1 hour. N,O-Dimethylhydroxylamine hydrochloride (1.5 g, 15.3 mmol) and Et3N (1.9 g, 19.1 mmol) were added and the mixture was stirred for an additional 5 hours. Ethyl acetate (100 mL) was added and the organic layer was washed with saturated aqueous NaHCO3 (400 mL × 3). The organic layer was separated, dried over Na2SO4, and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 2 / 1) to give the title compound (3.0 g, 85%) as a yellow solid. 1 H NMR (400 MHz, CDCl3): δ 8.50 (s, 1H), 7.80 (s, 1H), 3.52 (s, 3H), 3.39 (s, 3H). LC-MS (m / z): 279.0 [M+H] + .

[0181] Step b. 1-(5-Bromo-2-chloropyridin-3-yl)ethan-1-one: To a solution of 5-bromo-2-chloro-N-methoxy-N-methylnicotinamide (2.6 g, 9.3 mmol) in tetrahydrofuran (30 mL) was added methylmagnesium bromide (3.4 mL, 10.2 mmol) dropwise at 0 °C. The mixture was stirred at room temperature for 2 hours. The reaction was quenched with saturated NH4Cl and water (200 mL) was added. The organic layer was extracted with ethyl acetate (200 mL), dried over Na2SO4, and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 15 / 1) to give the title compound (2.0 g, 92%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ 8.55 (s, 1H), 8.02 (s, 1H), 2.70 (s, 3H). LC-MS (m / z): 234.0 [M+H] + .

[0182] Step c. 6-Bromo-1-(4-methoxybenzyl)-1,8-naphthyridin-4(1H)-one: 1-(5-Bromo-2-chloropyridin-3-yl)ethan-1-one (2.00 g, 8.55 mmol) was added to DMF-DMA (10 mL), and the mixture was stirred at 110 °C for 3 h. The solvent was removed in vacuo, and the residue was dissolved in N,N-dimethylformamide (10 mL). PMB-NH2 (1.76 g, 12.82 mmol) and Cs2CO3 (5.57 g, 17.09 mmol) were added, and the mixture was stirred at 110 °C overnight. Ethyl acetate (100 mL) was added, and the organic layer was washed with saturated aqueous NaCl solution (400 mL × 3). The organic layer was separated, dried over Na2SO4, and concentrated. The residue was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 120 / 1) to give the title compound (700 mg, 24%) as a yellow solid. 1 1H NMR (400 MHz, CDCl3) δ 8.81 (d, J = 2.4 Hz, 1H), 8.75 (d, J = 2.4 Hz, 1H), 7.72 (d, J = 8.0 Hz, 1H), 7.23 - 7.20 (m, 2H), 6.87 - 6.85 (m, 2H), 6.33 (d, J = 7.6 Hz, 1H), 5.48 (s, 2H), 3.78 (s, 3H). LC-MS (m / z): 345.1 [M+H] + .

[0183] Step d. 6-Bromo-1,8-naphthyridin-4-ol: 6-Bromo-1-(4-methoxybenzyl)-1,8-naphthyridin-4(1H)-one (700 mg, 0.03 mmol) was added to TFA (2 mL). The mixture was stirred at 110 °C for 2 h under microwave irradiation. The solvent was removed, and the pH was adjusted to 7 using saturated NaHCO3. Water (100 mL) was added. The organic layer was extracted with dichloromethane / methanol (50 mL / 10 mL), dried over Na2SO4, and concentrated. The residue was purified by silica gel column chromatography (dichloromethane / methane (v / v) = 50 / 1) to give the title compound (320 mg, 70%) as a yellow solid. 11H NMR (400 MHz, DMSO-d6) δ 12.37 (s, 1H), 8.85 (s, 1H), 8.53 (s, 1H), 8.02 - 7.96 (m, 1H), 6.15 (d, J = 6.8 Hz, 1H). LC-MS (m / z): 224.9 [M+H] + .

[0184] Step e. 3-Bromo-5-chloro-1,8-naphthyridine: 6-Bromo-1,8-naphthyridin-4-ol (300 mg, 1.33 mmol) was added to POCl3 (4 mL). The mixture was stirred at 110 °C for 1 h. The solvent was removed in vacuo to give the crude product (350 mg) as a brown solid of the title compound.

[0185] Step f. N-(6-Bromo-1,8-naphthyridin-4-yl)benzo[d]thiazol-5-amine: To a solution of 3-bromo-5-chloro-1,8-naphthyridine (350 mg, 1.44 mmol) in ethanol (3 mL) were added benzo[d]thiazol-5-amine (237 mg, 1.58 mmol) and concentrated hydrochloric acid (1 drop). The mixture was stirred at 80 °C for 3 h. Water (30 mL) was added and the pH was adjusted to ~7 with saturated NaHCO3. The organic layer was extracted with dichloromethane / methanol (30 mL / 10 mL × 2), combined, dried over Na2SO4, and concentrated. The residue was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 30 / 1) to give the title compound (105 mg, 20%) as a yellow solid. 1 1H NMR (400 MHz, DMSO-d6) δ 11.78 (s, 1H), 9.81 (s, 1H), 9.55 (s, 1H), 9.28 (s, 1H), 8.55 (d, J = 5.6 Hz, 1H), 8.40 (d, J = 8.0 Hz, 1H), 8.22 (s, 1H), 7.62 (d, J = 8.4 Hz, 1H), 6.96 (d, J = 6.4 Hz, 1H). LC-MS (m / z): 357.0 [M+H] + .

[0186] Step g. (5-(Benzo[d]thiazol-5-ylamino)-1,8-naphthyridin-3-yl)dimethylphosphine oxide: To a solution of N-(6-bromo-1,8-naphthyridin-4-yl)benzo[d]thiazol-5-amine (105 mg, 0.29 mmol) in 1,4-dioxane (4 mL) were added dimethylphosphine oxide (34 mg, 0.44 mmol), Et3N (45 mg, 0.44 mmol), Pd2dba3 (27 mg, 0.03 mmol) and Xantphos (17 mg, 0.03 mmol). The mixture was stirred at 125 °C for 2 h under microwave irradiation. Saturated aqueous NaHCO3 was added to the solution and the solvent was stirred for an additional 30 min. The solvent was removed in vacuo and the residue was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 20 / 1) to give the title compound (10 mg, 10%).

[0187] Example 10. Method J

[0188] Synthesis of (4-(Benzo[d]thiazol-5-ylamino)-7-methoxysinnolin-6-yl)dimethylphosphine oxide (B2)

Chemical formula

[0189] Step a. 2-Amino-5-bromo-4-methoxybenzonitrile: To a solution of 2-amino-4-methoxybenzonitrile (2.0 g, 13.5 mmol) in acetonitrile (30 mL) was added NBS (2.7 g, 14.9 mmol) in acetonitrile (10 mL) at 0 °C. The mixture was stirred at room temperature for 30 min. The solvent was concentrated and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10 / 1) to give the title compound (2.5 g, 82%) as a yellow solid. 1 1H NMR (400 MHz, CDCl3) δ 7.52 (s, 1H), 6.23 (s, 1H), 4.48 (s, 2H), 3.89 (s, 3H). LC-MS (m / z): 228.0 [M+H] + .

[0190] Step b. 1-(2-Amino-5-bromo-4-methoxyphenyl)ethan-1-one: Methylmagnesium bromide (20.5, 61.5 mmol) was added to a two-necked flask. 2-Amino-5-bromo-4-methoxybenzonitrile in tetrahydrofuran (10 mL) was added dropwise to the solution at 0 °C. The mixture was stirred at room temperature for 5 minutes under a nitrogen atmosphere and further stirred at 55 °C for 16 hours. 6N hydrochloric acid solution was added to the reaction solution and stirred at room temperature for 50 minutes. HCl was neutralized with saturated NaHCO3 solution. The organic layer was extracted with dichloromethane (50 mL × 3), combined, dried over Na2SO4, and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 5 / 1) to obtain the desired product (700 mg, 33%) as a gray solid. 1 1H NMR (400 MHz, CDCl3) δ 7.83 (s, 1H), 6.45 (s, 2H), 6.08 (s, 1H), 3.87 (s, 3H), 2.51 (s, 3H). LC-MS (m / z): 244.0 [M + H] + .

[0191] Step c. 6-Bromo-7-methoxysinnolin-4-ol: 1-(2-Amino-5-bromo-4-methoxyphenyl)ethan-1-one (700 mg, 2.9 mmol) was added to concentrated hydrochloric acid (10 mL) at 0 °C and stirred at this temperature for 15 minutes. NaNO2 (221 mg, 3.2 mmol) in water (1 mL) was added to the solution, and the mixture was stirred at 0 °C for 1.5 hours, at room temperature overnight, and at reflux for 6 hours. Hydrochloric acid was neutralized with saturated aqueous NaHCO3. The organic layer was extracted with dichloromethane / methane (35 mL / 7 mL × 5), dried over Na2SO4, and concentrated. The residue was purified by silica gel column chromatography (dichloromethane / methane (v / v) = 50 / 1) to obtain the target product (350 mg, 69%) as a white solid. 1 1H NMR (400 MHz, DMSO-d6) δ 13.43 (s, 1H), 8.15 (s, 1H), 7.73 (s, 1H), 7.01 (s, 1H), 3.98 (s, 3H). LC-MS (m / z): 255.0 [M + H] + .

[0192] Step d. N-(6-Bromo-7-methoxysinnolin-4-yl)benzo[d]thiazol-5-amine hydrochloride: 6-Bromo-7-methoxysinnolin-4-ol (350 mg, 1.4 mmol) was added to POCl3 (4 mL). The mixture was stirred at 110 °C for 2 h. The solvent was concentrated and dissolved in ethanol (5 mL). Benzo[d]thiazol-5-amine (225 mg, 1.5 mmol) was added to the reaction solution, and the mixture was stirred at 80 °C for 2 h. The obtained solid was collected by filtration, washed with ethanol and dried in vacuo to give the title compound (290 mg, 54%) as the hydrochloride salt of a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 15.79 (s, 1H), 12.18 (s, 1H), 9.54 (s, 1H), 9.37 (d, J = 10.4 Hz, 1H), 8.54 (s, 1H), 8.39 (d, J = 8.0 Hz, 1H), 8.30 (s, 1H), 7.68 (d, J = 8.4 Hz, 1H), 7.54 (d, J = 6.4 Hz, 1H), 4.11 (s, 3H). LC-MS (m / z): 386.9 [M + H] + .

[0193] Step e. (4-(Benzo[d]thiazol-5-ylamino)-7-methoxysinnolin-6-yl)dimethylphosphine oxide: To a solution of N-(6-bromo-7-methoxysinnolin-4-yl)benzo[d]thiazol-5-amine hydrochloride (130 mg, 0.34 mmol) in 1,4-dioxane (4 mL) were added dimethylphosphine oxide (52 mg, 0.68 mmol), Et3N (103 mg, 1.02 mmol), Pd2dba3 (31 mg, 0.03 mmol) and Xantphos (17 mg, 0.03 mmol). The mixture was stirred at 120 °C for 4 h under microwave irradiation. Saturated aqueous NaHCO3 was added to the solution, and the solvent was stirred for a further 30 min. The solvent was removed in vacuo, and the residue was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 20 / 1) to give the crude product. This was washed with ethanol / ether (1 mL / 5 mL) to give the desired product (25 mg, 19%) as a yellow solid.

[0194] Table 1 shows the selection of the compounds shown in the third column of the table, prepared according to the method described in detail above.

[0195] Table 1. Selected Compounds of the Invention of the Present Application (A1 - A29, B1 - B2). [Table 1] JPEG0007712515000048.jpg177135JPEG0007712515000049.jpg185134JPEG0007712515000050.jpg154135JPEG0007712515000051.jpg162135JPEG0007712515000052.jpg209140

[0196] Example 11. Binding Affinity Assay

[0197] In most assays, kinase-tagged T7 phage strains were prepared in an E. coli host derived from the BL21 strain. The E. coli was grown to the logarithmic phase, infected with T7 phage, and cultured with shaking at 32 °C until lysis. The lysate was centrifuged and filtered to remove cell debris. The remaining kinase was produced in HEK-293 cells and then tagged with DNA for qPCR detection. Magnetic beads coated with streptavidin were treated with biotinylated small molecule ligands at room temperature for 30 minutes to prepare affinity resin for kinase assay. The ligand beads were blocked with excess biotin, washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween20, 1 mM DTT) to remove unbound ligand, and to reduce non-specific binding. The binding reaction was constituted by binding kinase, ligand affinity beads, and test compound in 1x binding buffer (20% SeaBlock, 0.17x PBS, 0.05% Tween20, 6 mM DTT). The test compound was prepared as an 111-fold stock in 100% DMSO. Kd was determined using an 11-point three-fold dilution series containing three DMSO control points. Compounds for Kd measurement were all dispensed by acoustic transfer (non-contact dispensing) in 100% DMSO. Then, the compounds were directly diluted into the assay such that the final concentration of DMSO was 0.9%. All reactions were carried out in polypropylene 384-well plates. The final volume of each was 0.02 ml. The assay plates were incubated with shaking at room temperature for 1 hour, and the affinity beads were washed with wash buffer (1x PBS, 0.05% Tween 20). Then, the beads were resuspended in elution buffer (1x PBS, 0.05% Tween 20, 0.5 μM non-biotinylated affinity ligand) and incubated with shaking at room temperature for 30 minutes. The kinase concentration in the eluate was measured by qPCR.

[0198] 11-point three-fold serial dilutions of each test compound were prepared in 100% DMSO at 100-fold the final test concentration and then diluted 1-fold in the assay (final DMSO concentration = 1%). Most Kds were determined using the highest concentration of the compound = 30,000 nM. If the first Kd determined was less than 0.5 nM (the lowest concentration tested), the measurement was repeated with serial dilutions starting from a lower highest concentration. A Kd value reported as 40,000 nM indicates that the Kd was determined to be >30,000 nM.

[0199] Binding constants (Kds) were calculated with a standard dose-response curve using Hill's equation:

Equation

[0200] The Hill Slope was set to -1.

[0201] Curves were fitted using non-linear least squares fitting with the Levenberg-Marquardt algorithm.

[0202] Table 2. Binding affinities of test compounds for RIP2 kinase

Table 2

[0203] Conclusion: As shown in Table 2, compounds A1-A16, A20-A21, A24-A26 showed high affinity for RIP2 kinase, and B1 showed low affinity.

[0204] Example 12. THP-1 cell assay

[0205] Acute monoblastic and monocytic leukemia (THP-1) was obtained from ATCC (Catalog No.: TIB-202 TM)。The temperature of the sterile incubator is 37 - 38 °C, the osmotic pressure is 260 - 320 mmol / L, the pH is 7.2 - 7.4, and the carbon dioxide ratio is 5%.

[0206] The cells were seeded in a 96-well plate. After 1 hour, the cells were incubated with the test compound for 2 hours. MDP (10 μL) was added and incubated for 6 hours. The samples were centrifuged for 5 minutes (3000 rmp / min). The IL-8 concentration in the supernatant was detected by IL-8 ELISA.

[0207] Table 3. IL-8 inhibitory effect of test compounds (10 nM)

Table 3

[0208] Conclusion: From Table 3, compounds A6 - A11, A17 - A19, A22 - A23, and A27 - 29 effectively inhibited the IL-8 of THP-1 induced by MDP, while B2 may have no effect.

[0209] Example 13. Thermodynamic solubility test

[0210] Experimental procedure a. Add assay buffer to the compound powder to make a 4 mg / mL solution. b. Shake the sample tube for 1 hour (1000 rpm) and equilibrate overnight at room temperature. c. Centrifuge the sample (10 minutes - 12000 rpm) to precipitate undissolved particles. d. Transfer the supernatant to a new tube. e. Measure the concentration of the supernatant after centrifugation by LCMSMS detection.

[0211] Table 4. Thermodynamic solubility of test compounds

Table 4

[0212] Conclusion: Table 4 shows that compounds A6 - A9 and A13 have good solubility.

[0213] Example 14. CYP Inhibition Test

[0214] Experimental Procedure

[0215] Preheat 0.1M potassium phosphate buffer (K buffer), pH 7.4:

[0216] 100 mM K - buffer: Mix 9.5 mL of stock A with 40.5 mL of stock B, make up the total volume to 500 mL with Milli - Q water, and titrate to pH 7.4 with KOH or H3PO4.

[0217] Stock A (1M monobasic potassium phosphate): Dissolve 136.5 g of monobasic potassium phosphate in 1 L of Milli - Q water.

[0218] Stock B (1M dibasic potassium phosphate): Dissolve 174.2 g of dibasic potassium phosphate in 1 L of Milli - Q water.

[0219] Prepare test compounds and reference inhibitors (400×) in a 96 - well plate: a. Transfer 8 μL of 10 mM test compound to 12 μL of ACN. b. Prepare inhibitors of CYP1A2, CYP2C9 and CYP2D6 as a cocktail: 12 μL of 1 mM α - Naphthoflavon+10 μL of 40 mM sulfaphenazole+10 μL of 10 mM quinidine+8 μL of DMSO. c. Prepare inhibitor spike solutions for CYP3A4, CYP2B6, CYP2C8 and CYP2C19.

[0220] Prepare 4×NADPH cofactor (66.7 mg NADPH, 10 mL of 0.1M K buffer, pH 7.4).

[0221] As shown in the following table, prepare 4×substrates (2 mL for each isoform) (add HLM on ice if necessary).

[0222] Prepare 0.2 mg / mL HLM solution (10 μL of 20 mg / mL and 990 μL of 0.1 M K-buffer).

[0223] Add 400 μL of 0.2 mg / mL HLM to the assay well, and then add 2 μL of 400x test compound (see Step 2.1) to the designated well (see Table 1) on ice.

[0224] Add 200 μL of 0.2 mg / mL HLM to the assay well, and then add 1 μL of reference inhibitor solution (see Steps 2.2 and 2.3) to the designated well (see Table 1) on ice.

[0225] Add the following solutions to the 96-well assay plate on ice: a. Add 30 μL of 2x test compound and reference compound to the 0.2 mg / mL HLM solution (see Steps 6 and 7); b. Add 15 μL of 4x substrate solution (see Step 4).

[0226] Pre-incubate the 96-well assay plate and NADPH solution at 37 °C for 5 minutes.

[0227] Add 15 μL of pre-warmed 8 mM NADPH solution to the assay plate to initiate the reaction (see Step 3).

[0228] Incubate the assay plate at 37 °C. For 3A4 for 5 minutes, for 1A2, 2B6, 2C8, 2C9 and 2D6 for 10 minutes, and for 2C19 for 45 minutes.

[0229] Stop the reaction by adding 120 μL of ACN containing IS (see IS preparation in Table 2).

[0230] After quenching, the plate was shaken for 10 minutes (600 rpm / min) with a vibrator (IKA, MTS 2 / 4), and then centrifuged at 5594 g for 15 minutes (Thermo Multifuge × 3R).

[0231] Transfer 50 μL of the supernatant from each well to a 96-well sample plate containing 50 μL of ultrapure water (Millipore, ZMQS50F01) for LC / MS analysis.

[0232] Table 5. System for CYP inhibition

[0233] Refer to Figure 4.

[0234] Table 6. CYP inhibitory effects of test compounds

Table 5

[0235] Conclusion: As shown in Table 6, compounds A6, A14 - A16 (10 μM) have no obvious effect on CYP isoenzymes. The favorable inhibition suggests low drug / drug interactions of these compounds.

[0236] Example 15. Caco-2 permeability test

[0237] Experimental procedure 1. Pre-warm the HBSS buffer in a 37°C water bath. 2. Take out the compound from -20°C and sonicate it for several minutes (1 minute or more). 3. Solution preparation

[0238] For the A-to-B direction: · HBSS buffer containing 0.3% DMSO and 5 μM LY: Add 150 μL of DMSO and 50 μL of LY (5 mM) to 50 ml of HBSS buffer (pH 7.4). · HBSS buffer containing 0.1% DMSO and 5 μM LY: Add 50 μL of DMSO and 50 μL of LY (5 mM) to 50 mL of HBSS buffer (pH 7.4).

[0239] For the B-to-A direction: · HBSS buffer with 0.3% DMSO: Add 150 μL of DMSO to 50 mL of HBSS buffer (pH 7.4). · HBSS buffer with 0.1% DMSO: Add 50 μL of DMSO to 50 mL of HBSS buffer (pH 7.4).

[0240] Receiver solution buffer:

[0241] For the A-to-B direction:

[0242] Prepare HBSS buffer containing 0.4% DMSO: Add 200 μL of DMSO to 50 mL of HBSS buffer (pH 7.4).

[0243] For the B-to-A direction: · Prepare HBSS buffer containing 0.4% DMSO and 5 μM LY: Add 200 μL of DMSO and 50 μL of LY (5 mM) to 50 mL of HBSS buffer (pH 7.4).

[0244] Table 7. Preparation of donor solution

[0245] Refer to Figure 5. · Remove the cell culture plate from the incubator, wash the cell monolayer with HBSS buffer, and then measure the TEER value at room temperature. · Before loading into the donor chamber, centrifuge the compound solution (from step 3) at 4000 rpm for 5 minutes. · Add the solution based on the volumes described in the following table (confirm that an extra 100 μL of donor sample for T0 is taken as a backup).

[0246] Table 8. Solution volume [Table 6] ·7. To measure the LY concentration in the apical chamber, collect 100 μL of sample from the apical chamber onto an opaque plate for LYT0. ·8. After preheating the apical plate and the basolateral plate at 37 °C for about 5 minutes, place the apical plate on the basolateral plate and start the transport. ·9. Leave the plate standing in an incubator at 37 °C for 90 minutes. ·10. Prepare the 20× solution.

[0247] Table 9. Preparation of the working solution

Table 7

[0248] Table 10. Caco-2 permeability of the test compounds

Table 8

[0249] Conclusion: As can be seen from Table 10, Compounds A6 and A24 had high permeability and no obvious efflux. Compound A8 also showed good permeability but was an efflux substrate. Compounds A9, A14, and A15 had moderate permeability, while Compounds A7, A13, and A16 had low permeability. It can be seen that a slight change in the substituent greatly changes the permeability and causes efflux.

[0250] Example 16. Protein binding test

[0251] Experimental procedure

[0252] Spiking solutions of test compounds and reference compounds 1.1 Solution A (0.5 mM): Add 10 μL of a 10 mM stock solution to 190 μL of DMSO. 1.2 Solution B (0.02 mM): Add 8 μL of Solution A to 192 μL of 0.05 M sodium phosphate buffer. The final DMSO concentration of Solution B is 4%.

[0253] 2. Preparation of test compounds and reference compounds 2.1 Pre-load 380 μL aliquots of plasma into the wells designated for plasma and buffer respectively in a 96-well plate. 2.2 Add 20 μL of Solution B (0.02 mM test compound and reference compound) to the plasma pre-set in the 96-well plate. The final test concentration is 1 μM containing 0.2% DMSO.

[0254] 3. Loading of dialysis samples 3.1 Preparation of plasma for buffer systems (duplicate): Apply 100 μL aliquots of blank dialysis buffer to the receiver side of the dialysis chamber. Next, inject 100 μL of plasma added with test compounds and reference compounds into the donor side of the dialysis cell. 3.2 Preparation of T0 plasma samples for initial concentration (duplicate): 3.2.1 Dispense 25 μL of plasma added with test substance and reference compound as T0 plasma samples into a 96-well sample preparation plate. 3.2.2 Mix with the same amount of blank buffer (50:50, v / v). 3.2.3 Quench the sample with 200 μL of acetonitrile containing internal standard (IS). 3.3 Cover the dialysis block with a plastic lid, place the whole device on a shaker (60 rpm), and leave it standing at 37 °C for 5 hours. 3.4 Preparation of dialysis samples after 5-hour incubation 3.4.1 Pipette 25 μL each from both the donor and receiver sides of the dialysis device into a new sample preparation plate, and mix with the same amount of the opposite matrix (blank buffer → plasma and vice versa). 3.4.2 Quench the samples with 200 μL of acetonitrile containing the internal standard (IS). Vortex all samples (0 h and 5 h) at 600 rpm for 10 min, then centrifuge at 5594 g for 15 min (Thermo Multifuge × 3R). 3.4.3 Transfer 50 μL of the supernatant to a new 96-well plate and mix with 50 μL of Milli-Q water. Cover the sample plate and store in a freezer (-20 °C) until LC / MS / MS analysis.

[0255] Table 11. Results of Protein Binding of Compounds

Table 9

[0256] Conclusion As shown in Table 11, compounds A6, A14 - A16 have moderate protein binding in humans, rats, and dogs, while compound A24 has high protein binding. Compound A25 has high protein binding in rats and moderate protein binding in humans and dogs.

[0257] Example 17. Metabolic Stability Test

[0258] Experimental Procedure

[0259] 1. Buffer:

[0260] Buffer A: 1.0 L of 0.1 M potassium monobasic phosphate buffer containing 1.0 mM EDTA

[0261] Buffer B: 1.0 L of 0.1 M potassium dibasic phosphate buffer containing 1.0 mM EDTA

[0262] Buffer C: 0.1 M potassium phosphate buffer, 1.0 mM EDTA, pH 7.4. While monitoring with a pH meter, titrate 700 mL of Buffer B with Buffer A.

[0263] 2. Spiking solutions for the reference compound (Ketanserin) and the test compound:

[0264] 500 μM spiking solution: Add 10 μL of a 10 mM DMSO stock solution to 190 μL of ACN.

[0265] 1.5 μM spiking solution (0.75 mg / mL): Add 1.5 μM of the 500 μM spiking solution and 18.75 μL of 20 mg / mL liver microsomes to 479.75 μL of Buffer C on ice.

[0266] 3. Dissolve NADPH in Buffer C to prepare a NADPH stock solution (6 mM).

[0267] 4. Dispense 30 μL of the 1.5 μM spiking solution containing 0.75 mg / mL microsome solution into the assay plates designated for each time point (0-, 5-, 15-, 30-, 45-min) on ice.

[0268] 5. At the 0-minute time point, add 135 μL of ACN containing IS to the wells of the 0-minute plate, and then add 15 μL of the NADPH stock solution (6 mM).

[0269] 6. Incubate all other plates at 37 °C for 5 minutes.

[0270] 7. Add 15 μL of the NADPH stock solution (6 mM) to the plate to start the reaction and timing.

[0271] 8. After 5 minutes, 15 minutes, 30 minutes, and 45 minutes, add 135 μL of ACN containing IS to the wells of the corresponding plates to stop the reaction.

[0272] 9. After quenching, shake the plate with a vibrator (IKA, MTS 2 / 4) for 10 minutes (600 rpm / min) and centrifuge at 5594 g for 15 minutes (Thermo Multifuge × 3R).

[0273] 10. Transfer 50 μL of the supernatant from each well to a 96-well sample plate containing 50 μL of ultrapure water (Millipore, ZMQS50F01) for LC / MS analysis.

[0274] Table 12. Metabolic Stability of Test Compounds

Table 10

[0275] Conclusion: Table 12 showed that Compounds A6 - A9 and A13 - A16 exhibited low to moderate clearance in humans, rats, and dogs.

[0276] Example 18. Pharmacokinetic Evaluation

[0277] Objective 1. Evaluation of the Pharmacokinetic Profile of Candidate Compounds in Mice

[0278] Experimental Procedure

[0279] The pharmacokinetic properties of the compounds in mice were tested according to a standard protocol. The candidate compounds were prepared as clear solutions for single intravenous injection (i.v.) and suspensions for oral administration (p.o.). The vehicle for intravenous administration was 5% DMSO + 95% saline, and the vehicle for oral administration was 0.5% CMCNa. In the experiment, 48 male mice and 24 female mice were used and administered intravenously at a dose of 2 mg / kg. Plasma samples were collected at 0 h (before dosing) and 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 24 h after dosing. Additionally, 24 mice were administered orally at 10 mg / kg. Plasma samples were collected at 0 h (before dosing) and 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h after dosing.

[0280] Blood samples were placed in tubes containing K2-EDTA and stored on ice until centrifugation. The blood samples were centrifuged at 6800 g for 6 minutes at 2-8°C within 1 hour after collection and stored frozen at approximately -80°C.

[0281] The analytical results were confirmed using quality control samples for within-assay variation. The accuracy of more than 66.7% of the quality control samples and 50% of all QC samples at each concentration level were between 80% and 120% of the known values.

[0282] A standard set of parameters including area under the curve (AUC(0-t) and AUC(0-∞)), elimination half-life (T1 / 2), maximum plasma concentration (Cmax), and time to maximum plasma concentration (Tmax) were calculated by the study investigator using the non-compartmental analysis module of the FDA-approved pharmacokinetic program Phoenix WinNonlin 7.0 (Pharsight, USA).

[0283] Table 13. Mouse PK of the test compound

Table 11

[0284] Conclusion: Compounds A6, A24, and A25 had excellent plasma exposure and bioavailability in mice.

[0285] Objective 2. Evaluation of the pharmacokinetic profile of candidate compounds in rats

[0286] Experimental procedure

[0287] The pharmacokinetic properties of the compound were tested according to a standard protocol. The candidate compound was prepared as a clear solution for single intravenous injection (i.v.) and a suspension for oral administration (p.o.). The vehicle for intravenous administration was 5% DMSO + 95% saline, and the vehicle for oral administration was 0.5% CMCNa. Nine male rats and three rats were used in the experiment and administered intravenously at a dose of 2 mg / kg. Plasma samples were collected at 0 h (before dosing) and 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, and 24 h after dosing. Three rats were orally administered 10 mg / kg, and three rats were orally administered 100 mg / kg. Plasma samples were collected at 0 h (before dosing) and 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after dosing.

[0288] Blood samples were placed in tubes containing K2-EDTA and stored on ice until centrifugation. Blood samples were centrifuged at 6800 g for 6 minutes at 2 - 8°C within 1 hour after collection and stored frozen at approximately -80°C.

[0289] The analysis results were confirmed using quality control samples for within-assay variation. More than 66.7% accuracy of the quality control samples and 50% of all QC samples at each concentration level were within 80 - 120% of the known values.

[0290] A standard set of parameters including area under the curve (AUC(0-t) and AUC(0-∞)), elimination half-life (T1 / 2), maximum plasma concentration (Cmax), and time to maximum plasma concentration (Tmax) was calculated by the study investigator using the non-compartmental analysis module of the FDA-approved pharmacokinetic program Phoenix WinNonlin 7.0 (Pharsight, USA).

[0291] Table 14. Rat PK of the Test Compound

Table 12

[0292] Conclusion: Compounds A6 and A24 had excellent plasma exposure and bioavailability in rats.

[0293] Objective 3. Evaluation of the pharmacokinetic profile of candidate compounds in beagle dogs

[0294] Experimental procedure

[0295] The pharmacokinetic properties of the compounds in beagle dogs were tested according to the standard protocol. The candidate compounds were prepared as clear solutions for single intravenous injection (i.v.) and suspensions for oral administration (p.o.). The vehicle for intravenous injection was 5% DMSO + 95% saline, and the vehicle for oral administration was 0.5% CMCNa. Nine dogs were used in the experiment. 1 mg / kg was administered intravenously. Plasma samples were collected at 0 h (before dosing) and 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 24 h after dosing. 5 mg / kg was administered orally to 3 dogs. Another 3 dogs were administered 15 mg / kg or 30 mg / kg orally. Plasma samples were collected at 0 h (before dosing) and 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h after dosing.

[0296] Blood samples were placed in tubes containing K2-EDTA and stored on ice until centrifugation. Blood samples were centrifuged at 6800 g for 6 minutes at 2-8°C within 1 hour after collection and stored frozen at approximately -80°C.

[0297] The analytical results were confirmed using quality control samples for within-assay variation. The accuracy of more than 66.7% of the quality control samples and 50% of all QC samples at each concentration level was 80-120% of the known values.

[0298] A standard set of parameters including the area under the curve (AUC(0-t) and AUC(0-∞)), elimination half-life (T1 / 2), maximum plasma concentration (Cmax), and time to maximum plasma concentration (Tmax) was calculated by the study personnel using the non-compartmental analysis module of the FDA-approved pharmacokinetic program Phoenix WinNonlin 7.0 (Pharsight, USA).

[0299] Beagle dog PK in Table 15.A6

Table 13

[0300] Beagle dog PK in Table 16.A24

Table 14

[0301] Conclusion: Compounds A6 and A24 showed excellent plasma exposure and bioavailability in Beagle dogs.

[0302] Example 19. In Vivo Pharmacological Study of MDP (Muramyl Dipeptide)-Induced Peritonitis

[0303] Objective: To evaluate the in vivo activity of RIP2 kinase inhibitors.

[0304] Experimental Procedure

[0305] Mice were divided into four groups: a normal group, a DMSO group, a positive control group (GSK2983559), and a compound group. Thirty minutes before MDP injection (100 μg / mouse, ip), either 10 mg / kg of GSK2983559 or 3, 10 mg / kg of the candidate compound was administered to the mice by gavage. Three hours after MDP administration, blood samples were collected from the ophthalmic vein after anesthesia. IL-6 concentration was detected by ELISA.

[0306] Results: The experimental results were as shown in Figures 2 and 3.

[0307] Conclusion: After MDP injection, the IL-6 level increased significantly compared with the normal group. It was suggested that the downstream inflammatory pathway was activated and the model was successfully established. The IL-6 level in each administration group was lower than that in the model group. Among them, compounds A6 and A24 could more effectively inhibit the IL-6 level than the positive control GSK2983559 at the same dosage. These two compounds could more effectively inhibit the activity of RIP2 kinase than GSK2983559.

Claims

1. A compound of formula (I): 【Chemical 1】 or a pharmaceutically acceptable salt, solvate, isotope-labeled derivative, stereoisomer or tautomer thereof, wherein 【Chemical 2】 is 【Chemical Formula 3】 ; R ２ is H; R ３ -L is methoxy, -OCD ３ 、-OCF ３ 、-OCHF ２ 、-OCH ２ CH ２ OH, -OCH ２ CH ２ OCH ３ 、 [Chemical Formula 4] being R ４ is methyl; R ５ is methyl; R ７ is H.

2. The compound according to claim 1, or a pharmaceutically acceptable salt, solvate, isotope-labeled derivative, stereoisomer or tautomer thereof, wherein 【Chemical Formula 5】 is 【Chemical Formula 6】 is.

3. The compound according to claim 1, or a pharmaceutically acceptable salt, solvate, isotope-labeled derivative, stereoisomer or tautomer thereof, wherein R ３ -L is methoxy, -OCD ３ , -OCF ３ , or -OCHF ２ and is as follows.

4. The compound according to claim 1, or a pharmaceutically acceptable salt, solvate, isotope-labeled derivative, stereoisomer, or tautomer thereof, wherein the compound is 【Chemical Formula 7】 selected from the group consisting of.

5. A pharmaceutical composition comprising a therapeutically effective amount of the compound according to any one of claims 1 - 4 or a pharmaceutically acceptable salt, solvate, isotope-labeled derivative, stereoisomer or tautomer thereof, and a pharmaceutically acceptable carrier.

6. A composition comprising: (i) the compound according to any one of claims 1 - 4, or a pharmaceutically acceptable salt, solvate, isotope-labeled derivative, stereoisomer or tautomer thereof, or the pharmaceutical composition according to claim 5; and (ii) at least one additional therapeutic agent selected from the group consisting of an anti-tumor agent, an agent for treating autoimmune diseases, an anti-neurodegenerative agent, an agent for treating metabolic diseases, and an agent for treating genetic diseases.

7. Use of a therapeutically effective amount of the compound according to any one of claims 1 - 4 or a pharmaceutically acceptable salt, solvate, isotope-labeled derivative, or the pharmaceutical composition according to claim 5, or the composition according to claim 6, in the manufacture of a therapeutic agent for a disease or disorder associated with the RIP2 receptor in a mammal suffering from a disease or disorder associated with the RIP2 receptor, wherein the disease or disorder associated with the RIP2 receptor is a systemic inflammatory response, an autoimmune disease, a tumor, cancer, a metabolic disease or a neurodegenerative disease.

8. Use of a compound according to any one of claims 1 - 4 or a pharmaceutically acceptable salt, solvate, isotopically labeled derivative, stereoisomer or tautomer thereof, or a pharmaceutical composition according to claim 5, or a composition according to claim 6, in the manufacture of a therapeutic agent for a disease or disorder associated with the RIP2 receptor in a mammal suffering from a disease or disorder associated with the RIP2 receptor, wherein the disease or disorder associated with the RIP2 receptor is uveitis, dermatitis, acute pneumonia, type 2 diabetes, arthritis, ulcerative colitis, Crohn's disease, early onset inflammatory bowel disease, extraintestinal inflammatory bowel disease, prevention of ischemia-reperfusion injury in solid organ transplantation, non-alcoholic steatohepatitis, autoimmune hepatitis, asthma, systemic lupus erythematosus, sarcoidosis, Wegener's granulomatosis, interstitial lung disease, pulmonary fibrosis, renal fibrosis, liver fibrosis, myocardial infarction, hypersensitivity pneumonia, ankylosing spondylitis, multiple sclerosis, systemic sclerosis, polymyositis, rheumatoid arthritis, myasthenia gravis, type 1 diabetes, glomerulonephritis, autoimmune thyroiditis, graft rejection, Crohn's disease, Blau syndrome, scleroderma, psoriasis, stomatitis, retinitis pigmentosa, proliferative vitreoretinopathy, Best vitelliform macular dystrophy, eczema, urticaria, vasculitis, eosinophilic fasciitis, wet age-related macular degeneration, dry age-related macular degeneration, diabetic retinopathy, retinopathy of prematurity (ROP), diabetic macular edema, uveitis, retinal vein occlusion, cystoid macular edema, glaucoma, Parkinson's disease, Alzheimer's disease, Huntington's disease, breast cancer, lung cancer, bladder cancer, pancreatic cancer, liver cancer, head and neck squamous cell carcinoma, thyroid cancer, sarcoma, osteosarcoma, desmoid tumor, melanoma, prostate cancer, colorectal cancer, ovarian cancer, cervical cancer, esophageal cancer, gastric cancer, myeloma, lymphoma, mantle cell lymphoma, cutaneous T cell lymphoma, chronic and non-progressive anemia, primary or essential thrombocythemia, leukemia, acute leukemia, chronic leukemia, lymphocytic leukemia, myeloid leukemia, myelodysplastic syndrome, myeloproliferative disorder, brain tumor, astrocytoma, medulloblastoma, schwannoma, primitive neuroectodermal tumor, or pituitary tumor.

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