NON-ATP / CATALYTIC SITE p38 MITOGEN ACTIVATED PROTEIN KINASE INHIBITORS

Abstract

To provide compounds that inhibit p38α MAPK protein, and methods of using the same, for treating or preventing diseases such as cancer or inflammatory diseases.SOLUTION: For example, a compound illustrated below is disclosed.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates generally to compounds that are inhibitors of p38 mitogen-activated protein kinase (MAPK) protein, and more particularly, but not exclusively, to compounds that inhibit p38α MAPK protein by binding to a pocket near the ED substrate docking site of p38α MAPK, and methods of using such compounds as treatments for disease. [Background technology]

[0002] p38 mitogen-activated protein kinase (MAPK) contributes to the pathogenesis of many diseases, but currently available p38 catalytic inhibitors (e.g., SB203580) have low efficacy and cause toxicity, likely due to loss of activity against non-inflammatory p38 isoforms (e.g., p38β) and p38α-dependent counterregulatory responses (e.g., MSK1 / 2). Therefore, there is a need in the art for new therapeutic agents and therapies that address the selective inhibition of p38α MAPK and selectively block specific p38α MAPK functions to maintain important counterregulatory and homeostatic functions, with applications for the treatment of inflammatory and neoplastic diseases. Summary of the Invention

[0003] The present disclosure provides a compound of formula A, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof: [ka] In Formula A, Ar and Ar 1is independently selected from monocyclic or polycyclic optionally substituted cycloalkyl, monocyclic or polycyclic optionally substituted heterocycloalkyl, monocyclic or polycyclic optionally substituted aryl, monocyclic or polycyclic optionally substituted arylalkyl, monocyclic or polycyclic optionally substituted heteroaryl, and monocyclic or polycyclic optionally substituted heteroarylalkyl; R 1 and R 2 are independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted alkylheteroaryl; R 1 and R 2 can be optionally linked to form a carbocyclic or heterocyclic ring, and L 1 and L 2 are independently a bond, -NR a -, -S-, -S(O)-, -S(O)2-, -O-, -CR a 2-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NR a -, -NR a C(O)-, -C(O)NR a SO2-, -SO2NR a C(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NR a -, -NR a C(O)O-, -S(O) t N(R a )-(t is 1 or 2), -N(R a )S(O) t-(t is 1 or 2), disubstituted alkyl, disubstituted heteroalkyl, disubstituted alkenyl, disubstituted alkynyl, disubstituted cycloalkyl, disubstituted heterocycloalkyl, disubstituted aryl, disubstituted arylalkyl, disubstituted heteroaryl, and disubstituted heteroarylalkyl, any optional substituent independently at each occurrence is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR a , -SR a , -OC(O)-R a , -SC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -C(O)SR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, -N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (t is 1 or 2), and PO3 (Ra )2 and R a is independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl. In some embodiments, the optional substituents are independently selected from R 3 , R 4 , R 5 , and R 6 are selected from, independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted alkylheteroaryl; R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 Any two of Ar can be optionally linked to form a carbocyclic or heterocyclic ring. 1 is an optionally substituted aryl ring or an optionally substituted heteroaryl ring. In some embodiments, Ar is a monocyclic or polycyclic optionally substituted aryl, a monocyclic or polycyclic optionally substituted arylalkyl, a monocyclic or polycyclic optionally substituted heteroaryl, or a monocyclic or polycyclic optionally substituted heteroarylalkyl. In some embodiments, Ar is a 5- or 6-membered optionally substituted aryl or a 5- or 6-membered optionally substituted heteroaryl, and L 1 and L 2is linked to Ar in a 1,2, 1,3, or 1,4 substitution pattern. In some embodiments, Ar is a 1,2-, 1,3-, or 1,4-disubstituted phenyl, pyridine, pyrimidine, pyrazine, or triazine. In some embodiments, Ar is a 1,2-, or 1,3-disubstituted furan, thiophene, pyrrole, thiazole, imidazole, oxazole, triazole, or pyrazole. In some embodiments, Ar is a 1,2-, 1,3-, 1,4-, 1,5-, 1,6-, 1,7-, or 1,8-disubstituted naphthalene, quinoline, isoquinoline, or quinazoline. In some embodiments, Ar is a 1,2-, 1,3-, 1,4-, 1,5-, 1,6-, or 1,7-disubstituted indole or imidazole. In some embodiments, L 1 is a linker selected from -CH2-, -C(CH3)2-, and -C(CH2CH2)-; L 2 is a linker selected from -NHCH2-, -CH2NH-, -NHCO-, -CONH-, -SON2NH-, and -NHSO2-, and -NR 1 R 2 teeth, [ka] Selected from R 7 is hydrogen or optionally substituted alkyl, and Ar 2 is a heterocycle.

[0004] In one aspect, the disclosure relates to a compound of Formula I, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof: [ka]

[0005] In formula I, R 1 , R 2 , R 3 , R 4 , R 5 , and R 6are each independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted alkylheteroaryl; and R 1 and R 2 can be optionally linked to form a carbocyclic or heterocyclic ring, and L 1 is a linker selected from -CH2-, -C(CH3)2-, and -C(CH2CH2)-; L 2 is a linker selected from -NHCH2-, -CH2NH-, -NHCO-, -CONH-, -SO2NH-, and -NHSO2-; Ar 1 is an optionally substituted aryl ring or an optionally substituted heteroaryl ring. In some embodiments, -NR of Formula I 1 R 2 The group is selected from: [ka] In the formula, R 7 is hydrogen or optionally substituted alkyl, and Ar 2 is a heterocycle. In some embodiments, —NR 1 R 2 The group is selected from the following: [ka] In some embodiments, L 1 is -CH-. In some embodiments, R 3 , R 4 , R 5 , and R 6 is hydrogen. In some embodiments, L 2 is selected from -NHCH2-, -NHCO-, and -NHSO2-. In some embodiments, Ar 1 is selected from: [ka] In the formula, R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , and R 13 each independently represents hydrogen, halogen, -NR 1 R 2 , alkoxy, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted alkylheteroaryl; R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , and R 13 Any two vicinal rings of Ar can be optionally linked to form a carbocyclic or heterocyclic ring. 1 is selected from: [ka]

[0006] In another aspect, the present disclosure relates to a compound of Formula II, which is any of the compounds of Formulas 1001-1180 disclosed in Table 1. [ka]

[0007] In another aspect, the present disclosure relates to compounds of any of formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), and 1087 (SF-7-044). [Table 1-1] [Table 1-2]

[0008] In another aspect, the disclosure relates to a compound of any of Formulas A, I, II, and 1001-1180, which is a p38α MAPK inhibitor. In another aspect, the disclosure relates to a compound of any of Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044), which is a p38α MAPK inhibitor. In some embodiments, the p38α MAPK inhibitor is a selective inhibitor of p38α MAPK.

[0009] In another aspect, the disclosure relates to a p38α MAPK inhibitor of any of Formula A, Formula I, Formula II, and Formulas 1001-1180, wherein the p38α MAPK inhibitor can bind to a pocket near the ED substrate docking site of p38α MAPK and is defined by at least residues R49, H107, L108, and K165 of p38α MAPK. In some embodiments, the binding pocket is defined by residues R49, H107, L108, M109, G110, A157, V158, E163, L164, and K165 of p38α MAPK. In some embodiments, the p38α MAPK inhibitor is a selective inhibitor of p38α MAPK.

[0010] In another aspect, the present disclosure relates to a p38α MAPK inhibitor of any of formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044), wherein the p38α MAPK inhibitor is capable of binding to a pocket near the ED substrate docking site of p38α MAPK, defined by at least residues R49, H107, L108, and K165 of p38α MAPK. In some embodiments, the binding pocket is defined by residues R49, H107, L108, M109, G110, A157, V158, E163, L164, and K165 of p38α MAPK. In some embodiments, the p38α MAPK inhibitor is a selective inhibitor of p38α MAPK.

[0011] In one aspect, the disclosure relates to a pharmaceutical composition comprising a compound of any of Formula A, Formula I, Formula II, and Formulas 1001-1180, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof, and a physiologically compatible carrier vehicle, wherein the amount of compound in the composition is a therapeutically effective amount for the treatment or prevention of a disease alleviated by inhibiting the activity of p38α MAPK. In another aspect, the present disclosure relates to a pharmaceutical composition comprising a compound of any of Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof, and a physiologically compatible carrier vehicle, wherein the amount of compound in the composition is a therapeutically effective amount for the treatment or prevention of a disease alleviated by inhibiting the activity of p38α MAPK. In some embodiments, the disease is cancer or an inflammatory disease. In some embodiments, the disease is selected from the group consisting of rheumatoid arthritis, cardiovascular disease, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), asthma, acute respiratory distress syndrome (ARDS), and acute lung injury (ALI).In some embodiments, the cancer is acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, cholangiocarcinoma, bladder cancer, brain cancer, breast cancer, bronchogenic carcinoma, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngeal carcinoma, cystadenocarcinoma, embryonal carcinoma, endothelioma, ependymoma, epithelial carcinoma, esophageal cancer, Ewing's tumor, fibrosarcoma, gastric cancer, glioblastoma multiforme, glioma, head and neck cancer. , hemangioblastoma, hepatocellular carcinoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendothelial sarcoma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteogenic sarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma , sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, gastric cancer, sweat gland carcinoma, synovium, testicular cancer, small cell lung cancer, pharyngeal cancer, uterine cancer, Wilms' tumor, blood cancer, acute erythroleukemia, acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monoblastic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute nonlymphocytic leukemia, acute promyelocytic leukemia, acute anaplastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin's lymphoma, polycythemia vera, and Waldenstrom's macroglobulinemia.

[0012] In one aspect, the disclosure relates to a method of treating or preventing a disease that is alleviated by inhibiting p38α MAPK protein in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a p38α MAPK inhibitor, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof, wherein the p38α MAPK inhibitor is a compound of any of Formulas A, Formula I, Formula II, and Formulas 1001-1180. In another aspect, the disclosure relates to a method of treating or preventing a disease that is alleviated by inhibiting p38α MAPK protein in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a p38α MAPK inhibitor, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof, wherein the p38α MAPK inhibitor is a compound of Formula A, Formula I, Formula II, and Formulas 1001-1180. 1), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044). In some embodiments, the p38α MAPK inhibitor is administered in dosage unit form. In some embodiments, the dosage unit comprises a physiologically compatible carrier medium. In some embodiments, the disease is cancer or an inflammatory disease. In some embodiments, the disease is selected from the group consisting of rheumatoid arthritis, cardiovascular disease, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), asthma, acute respiratory distress syndrome (ARDS), and acute lung injury (ALI).In some embodiments, the cancer is acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, cholangiocarcinoma, bladder cancer, brain cancer, breast cancer, bronchogenic carcinoma, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngeal carcinoma, cystadenocarcinoma, embryonal carcinoma, endothelioma, ependymoma, epithelial carcinoma, esophageal cancer, Ewing's tumor, fibrosarcoma, gastric cancer, glioblastoma multiforme, glioma, head and neck cancer. , hemangioblastoma, hepatocellular carcinoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendothelial sarcoma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteogenic sarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma In some embodiments, the compound is selected from the group consisting of sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, gastric cancer, sweat gland carcinoma, synovium, testicular cancer, small cell lung cancer, pharyngeal cancer, uterine cancer, Wilms' tumor, blood cancer, acute erythroleukemia, acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monoblastic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute nonlymphocytic leukemia, acute promyelocytic leukemia, acute anaplastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin's lymphoma, polycythemia vera, and Waldenstrom's macroglobulinemia. In some embodiments, the compound selectively inhibits p38α MAPK. In some embodiments, inhibition of p38α MAPK does not result in a loss of p38α-dependent counterregulatory responses. In some embodiments, the p38α-dependent counterregulatory responses involve mitogen- and stress-activated protein kinase-1 (MSK1) or MSK2. In some embodiments, inhibition of p38α MAPK stabilizes endothelial or epithelial barrier function. In some embodiments, inhibition of p38α MAPK reduces inflammation. In some embodiments, inhibition of p38α MAPK alleviates lung injury. In some embodiments, inhibition of p38α MAPK alleviates LPS-induced lung injury. In some embodiments, inhibition of p38α MAPK modulates leukocyte trafficking. In some embodiments, inhibition of p38α MAPK modulates cytokine expression.

[0013] In one aspect, the disclosure relates to a method of treating or preventing a disease that is alleviated by inhibiting p38α MAPK protein in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a composition comprising a compound of any of Formula A, Formula I, Formula II, and Formulas 1001-1180, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof, and a physiologically compatible carrier vehicle. In another aspect, the disclosure relates to a method of treating or preventing a disease alleviated by inhibiting p38α MAPK protein in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a composition comprising a compound of any of Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof, and a physiologically compatible carrier vehicle. In some embodiments, the composition is administered in a dosage unit form. In some embodiments, the dosage unit comprises a physiologically compatible carrier medium. In some embodiments, the disease is cancer or an inflammatory disease. In some embodiments, the disease is selected from the group consisting of rheumatoid arthritis, cardiovascular disease, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), asthma, acute respiratory distress syndrome (ARDS), and acute lung injury (ALI).In some embodiments, the cancer is acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, cholangiocarcinoma, bladder cancer, brain cancer, breast cancer, bronchogenic carcinoma, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngeal carcinoma, cystadenocarcinoma, embryonal carcinoma, endothelioma, ependymoma, epithelial carcinoma, esophageal cancer, Ewing's tumor, fibrosarcoma, gastric cancer, glioblastoma multiforme, glioma, head and neck cancer. , hemangioblastoma, hepatocellular carcinoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendothelial sarcoma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteogenic sarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma In some embodiments, the compound is selected from the group consisting of sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, gastric cancer, sweat gland carcinoma, synovium, testicular cancer, small cell lung cancer, pharyngeal cancer, uterine cancer, Wilms' tumor, blood cancer, acute erythroleukemia, acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monoblastic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute nonlymphocytic leukemia, acute promyelocytic leukemia, acute anaplastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin's lymphoma, polycythemia vera, and Waldenstrom's macroglobulinemia. In some embodiments, the compound selectively inhibits p38α MAPK. In some embodiments, inhibition of p38α MAPK does not result in a loss of p38α-dependent counterregulatory responses. In some embodiments, the p38α-dependent counterregulatory responses involve mitogen- and stress-activated protein kinase-1 (MSK1) or MSK2. In some embodiments, inhibition of p38α MAPK stabilizes endothelial or epithelial barrier function. In some embodiments, inhibition of p38α MAPK reduces inflammation. In some embodiments, inhibition of p38α MAPK alleviates lung injury. In some embodiments, inhibition of p38α MAPK alleviates LPS-induced lung injury. In some embodiments, inhibition of p38α MAPK modulates leukocyte trafficking. In some embodiments, inhibition of p38α MAPK modulates cytokine expression.

[0014] In one aspect, the disclosure relates to a therapeutically effective amount of a p38α MAPK inhibitor, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof, for use in the treatment and prevention of a disease alleviated by inhibiting p38α MAPK protein in a patient in need thereof, wherein the p38α MAPK inhibitor is a compound of any of Formula A, Formula I, Formula II, and Formulas 1001-1180. In another aspect, the disclosure relates to a therapeutically effective amount of a p38α MAPK inhibitor, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof, for use in the treatment or prevention of a disease alleviated by inhibiting p38α MAPK protein in a patient in need thereof, wherein the p38α MAPK inhibitor is any of compounds of formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), and 1087 (SF-7-044). In some embodiments, the p38α MAPK inhibitor is administered in a dosage unit form. In some embodiments, the dosage unit comprises a physiologically compatible carrier medium. In some embodiments, the disease is cancer or an inflammatory disease. In some embodiments, the disease is selected from the group consisting of rheumatoid arthritis, cardiovascular disease, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), asthma, acute respiratory distress syndrome (ARDS), and acute lung injury (ALI).In some embodiments, the cancer is acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, cholangiocarcinoma, bladder cancer, brain cancer, breast cancer, bronchogenic carcinoma, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngeal carcinoma, cystadenocarcinoma, embryonal carcinoma, endothelioma, ependymoma, epithelial carcinoma, esophageal cancer, Ewing's tumor, fibrosarcoma, gastric cancer, glioblastoma multiforme, glioma, head and neck cancer. , hemangioblastoma, hepatocellular carcinoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendothelial sarcoma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteogenic sarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma In some embodiments, the compound is selected from the group consisting of sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, gastric cancer, sweat gland carcinoma, synovium, testicular cancer, small cell lung cancer, pharyngeal cancer, uterine cancer, Wilms' tumor, blood cancer, acute erythroleukemia, acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monoblastic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute nonlymphocytic leukemia, acute promyelocytic leukemia, acute anaplastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin's lymphoma, polycythemia vera, and Waldenstrom's macroglobulinemia. In some embodiments, the compound selectively inhibits p38α MAPK. In some embodiments, inhibition of p38α MAPK does not result in a loss of p38α-dependent counterregulatory responses. In some embodiments, the p38α-dependent counterregulatory responses involve mitogen- and stress-activated protein kinase-1 (MSK1) or MSK2. In some embodiments, inhibition of p38α MAPK stabilizes endothelial or epithelial barrier function. In some embodiments, inhibition of p38α MAPK reduces inflammation. In some embodiments, inhibition of p38α MAPK alleviates lung injury. In some embodiments, inhibition of p38α MAPK alleviates LPS-induced lung injury. In some embodiments, inhibition of p38α MAPK modulates leukocyte trafficking. In some embodiments, inhibition of p38α MAPK modulates cytokine expression. [Brief explanation of the drawings]

[0015] The foregoing summary, as well as the following detailed description of embodiments of the present invention, will be better understood when read in conjunction with the appended figures and drawings.

[0016] [Figure 1] Design of substrate-selective p38 inhibitors. Figure 1a shows the structure of p38α, showing the CD, ED, DEF, and activation sites. Figure 1b shows a comparison between the p38α and β structures. The CD and ED sites are colored red and blue, and the CADD target is colored yellow. The sequences containing the CADD target on p38α and the corresponding site on p38β differ by only three of 10 amino acids (highlighted in yellow). Figure 1c shows the overlap of the CADD target structures in apo (PDB:1p38, green) and dually phosphorylated (PDB:3PY3, yellow) mouse p38α. Figure 1d shows an overview of the CADD screening strategy. Figure 1e shows a DSF screen of compounds added to recombinant p38α or ERK2 at 10, 25, 50, or 100 μM, with binding indicated by an increase in melting temperature. Compounds that bind to ERK2 and p38α are highlighted in yellow. Only those that bind to p38α are highlighted in blue. Figure 1f shows the chemical structures of UM60, UM101, and SB203580. [Figure 2]The biological effects of p38 inhibitors are shown in Figures 2a and 2b. Figures 2a and 2b show the effects of 10 μM SB203580 (SB) or the indicated concentrations of UM60 or UM101 on HMVECL permeability (Figure 2a) and IL-8-specific neutrophil TEM (Figure 2b). Cells were pretreated with DMSO or compound for 1 hour and then incubated with 10 ng / ml TNFα for 6 hours before the permeability assay (Figure 2a), or incubated at 39.5°C for 6 hours without additional stimulation before the TEM assay (Figure 2b). Figures 2c and 2d show mean ± SE. * indicates p<0.0001 (vs. DMSO), † indicates p<0.0001 (vs. SB), and ‡ indicates p<0.005 (vs. 37°C). Male CD1 mice were pretreated with 1 mg SB or 0.1–1 mg UM101 prior to intratracheal (it) instillation of 50 μg LPS and heat exposure. * indicates p<0.05 (vs. DMSO). [Figure 3]The biochemical effects of substrate-selective p38 inhibitors are shown. Figures 3a and 3b illustrate heat maps from RNASeq showing the IPA pathway inhibited by SB203580 alone or by SB203580 and UM101 (Figure 3a), as well as the IPA pathway inhibited only by UM101 (Figure 3b). Figure 3c shows the biochemical effects of substrate-selective p38 inhibitors on HeLa cells pretreated with 50 μM UM101 or 10 μM SB203580 (SB) for 30 min, then treated with anisomycin for 10–60 min and immunoblotted for phosphorylated MK2, Stat-1, and total p38. Figure 3d shows DSF analysis of UM101 and SB203580 (SB) binding to recombinant p38α and p38β. Mean values ​​± SE for four experiments. *, †, and § indicate p<0.0001 vs. p38α (with DMSO), vs. p38β (with DMSO), and vs. p38β (with SB203580), respectively. P<0.0001 by MANOVA for the difference between UM101 binding to p38α and p38β. Figure 3e shows DSF analysis of UM101 and SB203580 (SB) binding to recombinant wild-type p38α and mutant p38α harboring four mutations in the CADD target pocket. Mean ± SE of four experiments. * and † indicate p<0.0001 vs. wild-type (with DMSO) and vs. mutant (with DMSO), respectively. P<0.0001 by MANOVA for the difference between UM101 binding to wild-type and mutant p38α. Figures 3f-k show STD-NMR analysis of UM101 with p38α (Figures 3f and 3g), p38β (Figures 3h and 3i), and p38α mutants (Figures 3j and 3k). 1D spectra (Figures 3f, 3h, and 3j) and STD spectra (Figures 3g, 3i, and 3k) from the same samples are shown. Tentative peak assignments are shown in Figure 3f. The inset shows the structure of UM101 with proton labeling. [Figure 4]Preliminary analysis of IL-8 and IL-1β mRNA by qRT-PCR prior to RNASeq. HMVECLs were preincubated with 0.4% DMSO, 10 μM SB203580, or 100 μM UM101 for 1 hour, then stimulated with 10 ng / ml TNFα for 4 hours. Total RNA was harvested, reverse transcribed, and analyzed by qRT-PCR to obtain fold changes relative to unstimulated control cells calculated using the delta-delta method with GAPDH as the housekeeping gene. [Figure 5] Four-quadrant map of RNASeq analysis. Genes with at least 10 reads in one sample per set and at least a 2-fold increase in TNFα are shown. The major component indicates the direction of change in UM101 / SB203580-treated cells relative to DMSO-treated cells. [Figure 6] FragMap of p38α SILCS. The nonpolar map (green) shows the putative binding pocket, and the location of the ED site is indicated. Maps of hydrogen bond donors (blue) and acceptors (red) are shown. [Figure 7] Figure 1 illustrates compound UM101 overlaid on a SILCS FragMap, which shows the aromatic (purple), aliphatic (green), positively charged (cyan), hydrogen bond acceptor (red), and hydrogen bond donor (blue) functional groups on the p38α backbone in wireframe with contours at -1.0 kcal / mol. The side chains of the four residues mutated in our CADD site-disrupting mutant are shown. The spatial distribution of the FragMap indicates where each functional group makes a favorable contribution to binding. [Figure 8] This shows that UM101 and analogs inhibit thrombin-induced cell permeability. Endothelial cells were pretreated with or without 10 or 15 mM UM101, SF-6-222 (6222), or SF-7-009 (7009). Cell permeability was measured using transepithelial / transendothelial electrical resistance (TEER). [Figure 9]UM101 and analogs inhibit acute lung injury in a mouse model. Mice were pretreated with 1 mg of test compound intraperitoneally (ip) and then treated with 100 mg of LPS intratracheally (it) at 39°C to induce inflammation. Lung injury was assessed by measuring total protein in lung lavage fluid. [Figure 10] This shows that UM101 and analogs inhibit acute lung injury in a mouse model. All mice received 50 μg of LPS (it) and were maintained at 39°C for 24 hours. All treatments were 1 mg in 0.5 ml of DMSO and were administered 6 hours after LPS. Mean ± SM, n = 5. * p = 0.03 (vs. DMSO #1), † p = 0.16 (vs. DMSO #2). Lung injury was assessed by measuring total protein in lung lavage fluid. DETAILED DESCRIPTION OF THE INVENTION

[0017] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All patents and publications mentioned herein are incorporated by reference in their entirety.

[0018] definition As used herein, the terms "administer," "administration," or "administering" refer to (1) providing, giving, dispensing, and / or prescribing by or under the direction of either a medical professional or their authorized representative in accordance with this disclosure, and / or (2) injecting, ingesting, or consuming into a mammal in accordance with this disclosure.

[0019] The terms "co-administration," "co-administering," "administered in combination with," "administering in combination with," "simultaneous," and "concurrent," as used herein, encompass the administration of two or more active pharmaceutical ingredients to a subject such that both active pharmaceutical ingredients and / or their metabolites are present in the subject at the same time. Co-administration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or administration in a composition in which two or more active pharmaceutical ingredients are present. Co-administration in separate compositions and administration in a composition in which both agents are present are preferred.

[0020] The terms "active pharmaceutical ingredient" and "drug" include the p38α MAPK inhibitors described herein, more specifically, the p38α MAPK inhibitors described by Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044). The terms "active pharmaceutical ingredient" and "drug" may also include compounds described herein that bind to p38α MAPK protein, thereby modulating the activity of p38α MAPK protein.

[0021] The term "isostere" refers to a group or molecule whose chemical and / or physical properties are similar to those of another group or molecule. A "bioisostere" is a type of isostere, and refers to a group or molecule whose biological properties are similar to those of another group or molecule. For example, in the case of the p38α MAPK inhibitors described herein, the carboxylic acid can be replaced with one of the following bioisosteres in place of the carboxylic acid, including, but not limited to, alkyl ester (COOR), acylsulfonamide (CONR-S02R), hydroxamic acid (CONR-OH), hydroxamate (CONR-OR), tetrazole, hydroxyisoxazole, isoxazol-3-one, and sulfonamide (S02NR), where each R can independently represent hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0022] The term "in vivo" refers to events that take place inside a subject's body.

[0023] The term "in vitro" refers to events that take place outside a subject's body. In vitro assays include cell-based assays, in which live or dead cells are used, and can also include cell-free assays, in which no intact cells are used.

[0024] The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound or combination of compounds described herein sufficient to accomplish its intended purpose, including, but not limited to, disease treatment. A therapeutically effective amount may vary depending on the intended use (in vitro or in vivo), the subject and disease state being treated (e.g., the subject's weight, age, and sex), the severity of the disease state, the method of administration, etc., and can be readily determined by one of ordinary skill in the art. The term also applies to a dose that induces a specific response in target cells (e.g., reduced platelet adhesion and / or cell migration). The specific dose will vary depending on the particular compound selected, the dosing regimen followed, whether the compound is administered in combination with other compounds, the timing of administration, the tissue to which it is administered, and the physical delivery system in which the compound is delivered.

[0025] A "therapeutic benefit," as that term is used herein, encompasses a therapeutic benefit and / or a prophylactic benefit. A prophylactic benefit includes delaying or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof.

[0026] As used herein, the terms "treat," "treatment," and / or "treating" can refer to the management of a disease, disorder, or pathological condition, or a symptom thereof, with the intent to cure, ameliorate, stabilize, and / or control the disease, disorder, or pathological condition, or a symptom thereof. More specifically, with respect to the control of a disease, disorder, or pathological condition, "control" can include the absence of condition progression, as assessed by response to the methods recited herein, which response can be complete (e.g., the disease goes into remission) or partial (e.g., any symptoms associated with the condition are reduced or alleviated). As used herein, the terms "prevent," "preventing," and / or "prevention" can refer to reducing the risk of a disease, disorder, or pathological condition occurring.

[0027] As used herein, the terms "modulate" and "modulation" refer to a change in the biological activity of a biological molecule (e.g., a protein, gene, peptide, antibody, etc.), where such a change may be related to an increase in the biological activity of the biological molecule (e.g., increased activity, agonism, activation, expression, up-regulation, and / or increased expression) or a decrease in the biological activity (e.g., decreased activity, antagonism, suppression, deactivation, down-regulation, and / or decreased expression). For example, the compounds described herein may modulate (i.e., inhibit) p38α MAPK protein. In some embodiments, the compounds described herein may selectively modulate (i.e., selectively inhibit) p38α MAPK protein compared to other MAPK or p38 MAPK proteins. In some embodiments, the compounds described herein may selectively modulate (i.e., selectively inhibit) p38α MAPK protein compared to other MAPK or p38 MAPK proteins.

[0028] The terms "QD," "qd," or "qd" mean once a day, once a day, or once daily. The terms "BID," "bid," or "bid" mean twice a day, twice a day, or twice daily. The terms "TID," "tid.", or "tid" mean three times a day, three times a day, or three times daily. The terms "QID," "qid," or "qid" mean four times a day, four times a day, or four times daily.

[0029] The term "pharmaceutically acceptable salts" refers to salts derived from various organic and inorganic counterions known in the art. Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids. Preferred inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Preferred organic acids from which 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, and salicylic acid. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins. Specific examples include isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salt is selected from ammonium salts, potassium salts, sodium salts, calcium salts, and magnesium salts. The term "cocrystal" refers to a molecular complex derived from several cocrystal formers known in the art. Unlike salts, cocrystals typically do not involve hydrogen transfer between the cocrystal and the drug, but instead involve intermolecular interactions such as hydrogen bonding, aromatic ring stacking, or dispersion forces between the cocrystal former and the drug in the crystal structure.

[0030] A "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" or "physiologically compatible" carrier or carrier medium is intended to include any and all solvents, dispersion media, coating agents, antibacterial and antifungal agents, isotonic and absorption delaying agents, and inactive ingredients. The use of such pharmaceutically acceptable carriers or pharmaceutically acceptable excipients for active pharmaceutical ingredients is well known in the art. Except insofar as any conventional pharmaceutically acceptable carrier or pharmaceutically acceptable excipient is incompatible with the active pharmaceutical ingredient, its use in the therapeutic compositions of the present invention is contemplated. Additional active pharmaceutical ingredients, such as other drugs, can also be incorporated into the compositions and methods described.

[0031] "Prodrug" refers to a derivative of a compound described herein, the pharmacological action of which results from conversion to an active compound by chemical or metabolic processes in vivo. Prodrugs include compounds in which an amino acid residue, or a polypeptide chain of two or more (e.g., two, three, or four) amino acid residues, is covalently linked via an amide or ester bond to a free amino, hydroxyl, or carboxylic acid group of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044). Amino acid residues include, but are not limited to, the 20 naturally occurring amino acids, generally designated by one- or three-letter symbols, but also include, for example, 4-hydroxyproline, hydroxylysine, desmosine, isodesmosine, 3-methylhistidine, β-alanine, γ-aminobutyric acid, citrulline, homocysteine, homoserine, ornithine, and methionine sulfone. Additional types of prodrugs are also encompassed. For example, free carboxyl groups can be derivatized as amides or alkyl esters (e.g., methyl esters and acetoxymethyl esters). As used herein, prodrug esters include esters and carbonates formed by reacting one or more hydroxyls of the compounds of the present invention with alkyl-, alkoxy-, or aryl-substituted acylating agents using procedures known to those skilled in the art to produce acetates, pivalates, methyl carbonates, benzoates, and the like. As a further example, free hydroxyl groups may be derivatized using groups including, but not limited to, hemisuccinate, phosphate ester, dimethylaminoacetate, and phosphoryloxymethyloxycarbonyl, as reviewed in Advanced Drug Delivery Reviews, 1996, 19, 115.Carbamate prodrugs of hydroxyl and amino groups are also included, as are carbonate prodrugs, sulfonate prodrugs, sulfonate esters, and sulfate esters of hydroxyl groups. Free amines can also be derivatized to amides, sulfonamides, or phosphonamides. All of the prodrug moieties described can incorporate groups including, but not limited to, ether, amine, and carboxylic acid functionalities. Furthermore, any compound that can be converted in vivo to provide a bioactive agent (e.g., compounds of Formulas A, I, II, III, and IV) is a prodrug within the scope of the present invention. Various forms of prodrugs are well known in the art. Comprehensive descriptions of prodrugs and prodrug derivatives are found in: (a) The Practice of Medicinal Chemistry, Camille G. Wermuth et al., (Academic Press, 1996), (b) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985), and (c) A Textbook of Drug Design and Development, P. Krogsgaard-Larson and H. Bundgaard, eds., (Harwood Academic Publishers, 1991). In general, prodrugs may be designed to improve penetration of a drug across biological membranes in order to obtain improved drug absorption, extend the duration of action of a drug (sustained release of the parent drug from the prodrug, reduced first-pass metabolism of the drug), target drug action (e.g., organ or tumor targeting, lymphocyte targeting), alter or improve the water solubility of a drug (e.g., iv preparations and eye drops), improve topical drug delivery (e.g., dermal and ocular drug delivery), improve the chemical / enzymatic stability of a drug, or reduce off-target drug effects, and more generally, improve the therapeutic efficacy of the compounds utilized in the present invention.

[0032] Unless otherwise stated, chemical structures depicted herein are intended to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds in which one or more hydrogen atoms are replaced by deuterium or tritium, or in which one or more carbon atoms are replaced by 13 C- or 14 Compounds replaced by C-enriched carbons are within the scope of the present invention.

[0033] When ranges are used herein to describe physical or chemical properties, such as, for example, molecular weight or chemical formula, all combinations and subcombinations of ranges, and specific embodiments therein, are intended to be included. The use of the term "about" when referring to a numerical value or numerical range means that the referenced numerical value or numerical range is approximate within experimental variation (or within statistical experimental error), and thus the numerical value or numerical range may vary. The variation is typically 0% to 15%, or 0% to 10%, or 0% to 5% of the stated numerical value or numerical range. The term "comprising" (and related terms such as "comprise" or "comprises," or "having" or "including") includes embodiments thereof, such as, for example, any composition, method, or process embodiment that "consists of" or "consists essentially of" the described features.

[0034] "Alkyl" refers to an alkyl group containing no unsaturation and having 1 to 10 carbon atoms (e.g., (C 1~10 ) alkyl or C 1~10"Alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, having a substituted or unsubstituted alkyl group (alkyl). Whenever it appears herein, a numerical range such as "1 to 10" refers to each integer within the given range; for example, "1 to 10 carbon atoms" means that the alkyl group can consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms, although this definition is also intended to encompass occurrences of the term "alkyl" where no numerical range is specifically specified. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, isobutyl, tertiary butyl, pentyl, isopentyl, neopentyl, hexyl, septyl, octyl, nonyl, and decyl. The alkyl moiety may be attached to the remainder of the molecule by a single bond, such as, for example, methyl (Me), ethyl (Et), n-propyl (Pr), 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), and 3-methylhexyl. Unless otherwise stated in the specification, alkyl groups are independently selected from heteroalkyl, acylsulfonamido, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NRa )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0035] "Alkylaryl" refers to an -(alkyl)aryl radical, where the aryl and alkyl are as disclosed herein and are optionally substituted with one or more of the substituents described as suitable substituents for aryl and alkyl, respectively.

[0036] "Alkylhetaryl" refers to an -(alkyl)hetaryl radical, where the hetaryl and alkyl are as disclosed herein and are optionally substituted with one or more of the substituents described as suitable substituents for aryl and alkyl, respectively.

[0037] "Alkylheterocycloalkyl" refers to an -(alkyl)heterocyclic radical, where the alkyl and heterocycloalkyl are as disclosed herein and are optionally substituted with one or more of the substituents described as suitable substituents for heterocycloalkyl and alkyl, respectively.

[0038] An "alkene" moiety refers to a group consisting of at least two carbon atoms and at least one carbon-carbon double bond. An "alkyne" moiety refers to a group consisting of at least two carbon atoms and at least one carbon-carbon triple bond. The alkyl moiety may be branched, straight-chain, or cyclic, whether saturated or unsaturated.

[0039] "Alkenyl" refers to an alkyl group consisting solely of carbon and hydrogen atoms, containing at least one double bond, and having from 2 to 10 carbon atoms (i.e., (C 2~10 ) alkenyl or C 2~10 "Alkenyl" refers to a straight or branched hydrocarbon chain radical group having an alkyl group (e.g., alkylsulfonamido, alkylsulfonamido, alkylsulfonamido, aryl ... a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a, -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0040] "Alkenyl-cycloalkyl" refers to an -(alkenyl)cycloalkyl radical, in which the alkenyl and cycloalkyl are as disclosed herein and are optionally substituted with one or more of the substituents described as suitable substituents for alkenyl and cycloalkyl, respectively.

[0041] "Alkynyl" refers to an alkyl group consisting solely of carbon and hydrogen atoms, containing at least one triple bond, and having from 2 to 10 carbon atoms (i.e., (C 2~10 ) alkynyl or C 2~10"Alkynyl" refers to a straight or branched hydrocarbon chain radical group having an alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, acylsulfonamido, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR, -OR, -OR. Each time it appears herein, a numerical range such as "2 to 10" refers to each integer within the given range; for example, "2 to 10 carbon atoms" means that the alkynyl group can consist of 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms. The alkynyl can be attached to the remainder of the molecule by a single bond, for example, ethynyl, propynyl, butynyl, pentynyl, and hexynyl. Unless otherwise stated herein, the alkynyl group can be independently selected from alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, acylsulfonamido, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR, -OR, -OR. a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a)2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0042] "Alkynyl-cycloalkyl" refers to an -(alkynyl)cycloalkyl radical, in which the alkynyl and cycloalkyl are as disclosed herein and are optionally substituted with one or more of the substituents described as suitable substituents for alkynyl and cycloalkyl, respectively.

[0043] "Acyl sulfonamide" refers to the group -C(=O)NR a -S(=O)2R a Each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0044] "Carboxaldehyde" refers to the --(C.dbd.O)H radical.

[0045] "Carbonyl" refers to the group -C(=O)-. The carbonyl group can be substituted with the following exemplary substituents: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, acylsulfonamido, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -NR a-OR a -, -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0046] "Carboxyl" refers to the --(C.dbd.O)OH radical.

[0047] "Cyano" refers to the -CN radical.

[0048] "Cycloalkyl" refers to a monocyclic or polycyclic radical that contains only carbon and hydrogen and may be saturated or partially unsaturated. Cycloalkyl groups contain 3 to 10 ring atoms (i.e., (C 3~10 ) cycloalkyl or C 3~10cycloalkyl). Wherever it appears herein, a numerical range such as "3 to 10" refers to each integer within the given range; for example, "3 to 10 carbon atoms" means that the cycloalkyl group can consist of up to and including 10 carbon atoms, such as 3 carbon atoms. Illustrative examples of cycloalkyl groups include, but are not limited to, the following moieties: cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl, and the like. Unless stated otherwise in the specification, cycloalkyl groups are independently selected from alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, acylsulfonamido, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR a , -SR a , -S(O) t R a -(t is 1 or 2), -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t ORa (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0049] "Cycloalkyl-alkenyl" refers to a -(cycloalkyl)alkenyl radical, in which the cycloalkyl and alkenyl are as disclosed herein and are optionally substituted with one or more of the substituents described as suitable substituents for cycloalkyl and alkenyl, respectively.

[0050] "Cycloalkyl-heterocycloalkyl" refers to a -(cycloalkyl)heterocycloalkyl radical, where the cycloalkyl and heterocycloalkyl are as disclosed herein and are optionally substituted with one or more of the substituents described as suitable substituents for cycloalkyl and heterocycloalkyl, respectively.

[0051] "Cycloalkyl-heteroaryl" refers to a -(cycloalkyl)heteroaryl radical, where the cycloalkyl and heteroaryl are as disclosed herein and are optionally substituted with one or more of the substituents described as suitable substituents for cycloalkyl and heteroaryl, respectively.

[0052] The term "alkoxy" refers to an -O-alkyl group containing from 1 to 8 carbon atoms of a straight, branched, or cyclic configuration and combinations thereof, attached to the parent structure through an oxygen. Examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, and cyclohexyloxy. A "lower alkoxy" refers to an alkoxy group containing from 1 to 6 carbons.

[0053] The term "substituted alkoxy" refers to an alkoxy in which the alkyl moiety is substituted (i.e., -O-(substituted alkyl)). Unless otherwise stated in the specification, the alkyl portions of the alkoxy group can independently be alkyl, heteroalkyl, alkenyl, acylsulfonamido, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a(t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0054] The term "alkoxycarbonyl" refers to a group of the formula (alkoxy)(C=O)- attached through the carbonyl carbon, where the alkoxy group has the indicated number of carbon atoms. 1~6 An alkoxycarbonyl group is an alkoxy group having from 1 to 6 carbon atoms attached through its oxygen to a carbonyl linker. A "lower alkoxycarbonyl" refers to an alkoxycarbonyl group in which the alkoxy group is a lower alkoxy group.

[0055] The term "substituted alkoxycarbonyl" refers to the group (substituted alkyl)-OC(O)-, which is attached to the parent structure via a carbonyl functionality. Unless otherwise stated in the specification, the alkyl portion of the alkoxycarbonyl group is independently selected from alkyl, heteroalkyl, acylsulfonamido, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(Ra )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0056] "Acyl" refers to the groups (alkyl)-C(O)-, (aryl)-C(O)-, (heteroaryl)-C(O)-, (heteroalkyl)-C(O)-, and (heterocycloalkyl)-C(O)-, which are attached to the parent structure via a carbonyl functionality. When the R radical is heteroaryl or heterocycloalkyl, the heterocycle or chain atoms contribute to the total number of chain or ring atoms. Unless otherwise specified herein, the alkyl, aryl, or heteroaryl portions of the acyl group are independently alkyl, heteroalkyl, acylsulfonamido, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR a , -SR a , -S(O)t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0057] "Acyloxy" refers to the R(C=O)O- radical, where R is alkyl, aryl, heteroaryl, heteroalkyl, or heterocycloalkyl, as described herein. When the R radical is heteroaryl or heterocycloalkyl, the heterocycle or chain atoms contribute to the total number of chain or ring atoms. Unless otherwise stated in the specification, R of an acyloxy group is independently alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R ais independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0058] "Amino" or "amine" refers to the -N(R a )2 radical groups, each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl, unless otherwise stated in the specification. a ) Two groups have two R a When substituted, they can be combined with the nitrogen atom to form a 4-, 5-, 6-, or 7-membered ring. For example, —N(R a )2 is meant to include, but is not limited to, 1-pyrrolidinyl and 4-morpholinyl. Unless otherwise stated in the specification, amino groups may independently be alkyl, acylsulfonamido, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(Ra )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0059] The term "substituted amino" also refers to the group -NHR, respectively, as described above. d and N.R. d R d The term "N-oxide" refers to the N-oxide of the formula: N-oxides can be prepared by treating the corresponding amino group with, for example, hydrogen peroxide or m-chloroperbenzoic acid.

[0060] An "amide" or "amido" is a group of the formula -C(O)NR a R b or -NR a C(O)R b refers to a chemical moiety having an R a and R b is selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heterocycle (bonded through a ring carbon), each of which moieties can be optionally substituted. a R b R for amide a and R bcan optionally form a 4-membered ring, 5-membered ring, 6-membered ring, or 7-membered ring together with the nitrogen to which they are attached. Unless otherwise specified herein, the amide group is optionally substituted with one or more of the substituents described herein, independently, for alkyl, amino, cycloalkyl, aryl, heteroaryl, or heterocycloalkyl. The amide can be an amino acid or peptide molecule attached to the compounds disclosed herein, thereby forming a prodrug. The procedures and specific groups for making such amides are known to those skilled in the art and can be easily found in well-known sources, such as Greene and Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley & Sons, New York, NY, 1999 (incorporated herein in its entirety by reference).

[0061] "Aromatic" or "aryl" or "Ar" refers to a group of 6 to 10 ring atoms (e.g., C6-C8) with at least one ring having a conjugated pi-electron system that is carbocyclic (e.g., phenyl, fluorenyl, and naphthyl). 10 Aromatic or C6-C 10"aryl" refers to aromatic radicals, including aryls. Divalent radicals formed from substituted benzene derivatives and having a free valence on a ring atom are named as substituted phenylene radicals. Divalent radicals derived from monovalent polycyclic hydrocarbon radicals ending in "-yl" by removing one hydrogen atom from the carbon atom having the free valence are named by appending "-ydene" to the name of the corresponding monovalent radical; for example, a naphthyl group with two points of attachment is called a naphthylidene. Wherever they appear herein, a numerical range such as "6 to 10" refers to each integer within the given range; for example, "6 to 10 ring atoms" means that the aryl group can consist of six ring atoms, seven ring atoms, etc., up to and including 10 ring atoms. This term includes monocyclic or fused-ring polycyclic (i.e., rings sharing adjacent pairs of ring atoms) groups. Unless otherwise stated in the specification, aryl moieties are independently selected from alkyl, heteroalkyl, acylsulfonamido, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O)t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0062] "Aralkyl" or "arylalkyl" refers to an (aryl)alkyl-radical in which the aryl and alkyl are as disclosed herein and are optionally substituted with one or more of the substituents described as suitable substituents for the aryl and alkyl, respectively.

[0063] "Ester" refers to a chemical radical of the formula -COOR, where R is selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heterocycle (bonded through a ring carbon). Procedures and specific groups for making esters are known to those skilled in the art and can be readily found in prominent sources such as Greene and Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley & Sons, New York, NY, 1999, which is incorporated herein by reference in its entirety. Unless otherwise specified herein, ester groups are independently selected from alkyl, acylsulfonamido, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR a , -SR a , -S(O)t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0064] "Fluoroalkyl" refers to an alkyl radical, as defined above, that is substituted by one or more fluoro radicals, as defined above, for example, trifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc. The alkyl portion of the fluoroalkyl radical can be optionally substituted as defined above for an alkyl group.

[0065] The terms "halo," "halide," or alternatively "halogen" are intended to mean fluoro, chloro, bromo, or iodo. The terms "haloalkyl," "haloalkenyl," "haloalkynyl," and "haloalkoxy" include alkyl, alkenyl, alkynyl, and alkoxy structures that are substituted with one or more halo groups or combinations thereof. For example, the terms "fluoroalkyl" and "fluoroalkoxy" include haloalkyl and haloalkoxy groups, respectively, where the halo is fluorine.

[0066] "Heteroalkyl," "heteroalkenyl," and "heteroalkynyl" refer to optionally substituted alkyl, alkenyl, and alkynyl radicals having one or more skeletal atoms selected from atoms other than carbon, e.g., oxygen, nitrogen, sulfur, phosphorus, or combinations thereof. Numerical ranges may be given, e.g., C1-C4 heteroalkyl refers to the total chain length, in this example, 4 atoms in length. Heteroalkyl groups are independently selected from alkyl, heteroalkyl, alkenyl, alkynyl, acylsulfonamido, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo, trimethylsilanyl, -OR a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(Ra )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0067] "Heteroalkylaryl" refers to an -(heteroalkyl)aryl radical, where the heteroalkyl and aryl are as disclosed herein and are optionally substituted with one or more of the substituents described as suitable substituents for heteroalkyl and aryl, respectively.

[0068] "Heteroalkylheteroaryl" refers to a -(heteroalkyl)heteroaryl radical, where the heteroalkyl and heteroaryl are as disclosed herein and are optionally substituted with one or more of the substituents described as suitable substituents for heteroalkyl and heteroaryl, respectively.

[0069] "Heteroalkylheterocycloalkyl" refers to a -(heteroalkyl)heterocycloalkyl radical, where the heteroalkyl and heterocycloalkyl are as disclosed herein and are optionally substituted with one or more of the substituents described as suitable substituents for heteroalkyl and heterocycloalkyl, respectively.

[0070] "Heteroalkylcycloalkyl" refers to a -(heteroalkyl)cycloalkyl radical, where the heteroalkyl and cycloalkyl are as disclosed herein and are optionally substituted with one or more of the substituents described as suitable substituents for heteroalkyl and cycloalkyl, respectively.

[0071] "Heteroaryl" or "heteroaromatic" or "HetAr" refers to a 5- to 18-membered aromatic radical (e.g., C5-C6) containing one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur. 13Heteroaryl refers to a heteroaryl group (heteroaryl) and may be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system. Wherever it appears herein, a numerical range such as "5 to 18" refers to each integer within the given range; for example, "5 to 18 ring atoms" means that the heteroaryl group can consist of 5 ring atoms, 6 ring atoms, and so on, up to and including 18 ring atoms. Divalent radicals derived from monovalent heteroaryl radicals ending in "-yl" by removing one hydrogen atom from the atom bearing the free valence are named by appending "-yden" to the name of the corresponding monovalent radical; for example, a pyridyl group with two points of attachment is a pyridylidene. An N-containing "heteroaromatic" or "heteroaryl" moiety refers to an aromatic group in which at least one of the skeletal atoms of the ring is a nitrogen atom. Polycyclic heteroaryl groups can be fused or non-fused. The heteroatom(s) of the heteroaryl radical are optionally oxidized. One or more nitrogen atoms, if present, are optionally quaternized. Heteroaryl can be attached to the remainder of the molecule through any atom of the ring(s). Examples of heteroaryl include azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzoxazolyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl. benzofurazanyl, benzothiazolyl, benzothienyl (benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furazanyl, furanonyl, furo[3,2-c]pyridinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl , isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, isoxazol-3-one, 5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl, Phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazo Linyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, thiapyranyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,

[0033] Unless otherwise specified herein, heteroaryl moieties are independently selected from alkyl, acylsulfonamido, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo, trimethylsilanyl, -OR, a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0072] Substituted heteroaryl also includes ring systems substituted with one or more oxide (—O—) substituents, such as, for example, pyridinyl N-oxide.

[0073] "Heteroarylalkyl" refers to a moiety having an aryl moiety, as described herein, bound to an alkylene moiety, as described herein, where the bond to the rest of the molecule is through the alkylene group.

[0074] "Heterocycloalkyl" refers to a stable 3- to 18-membered non-aromatic cyclic radical containing 2 to 12 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, oxygen, and sulfur. Wherever it appears herein, a numerical range such as "3 to 18" refers to each integer within the given range; for example, "3 to 18 ring atoms" means that the heterocycloalkyl group can consist of 3 ring atoms, 4 ring atoms, and so on, up to and including 18 ring atoms. Unless otherwise stated herein, a heterocycloalkyl radical is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems. The heteroatoms of a heterocycloalkyl radical can be optionally oxidized. One or more nitrogen atoms, if present, are optionally quaternized. A heterocycloalkyl radical is partially or fully saturated. A heterocycloalkyl can be attached to the remainder of the molecule through any atom of the ring(s). Examples of such heterocycloalkyl radicals include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless otherwise stated in the specification, heterocycloalkyl moieties are independently selected from alkyl, acylsulfonamido, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamate, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo, trimethylsilanyl, -OR a , -SR a , -S(O) t R a -(t is 1 or 2), -OC(O)-Ra , -N(R a )2, -C(O)R a , -C(O)OR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or -PO(OR a )2, and each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0075] "Heterocycloalkyl" also includes bicyclic ring systems in which one non-aromatic ring typically has 3 to 7 ring atoms and includes at least 2 carbon atoms, as well as 1 to 3 heteroatoms independently selected from oxygen, sulfur, and nitrogen, and a combination including at least one of the foregoing heteroatoms, and the other ring typically has 3 to 7 ring atoms and optionally includes 1 to 3 heteroatoms independently selected from oxygen, sulfur, and nitrogen, and is not aromatic.

[0076] "Hydroxamate" is -C(O)NR aOR a Each R refers to a part a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0077] "Nitro" refers to the -NO2 radical.

[0078] "Oxa" refers to the -O- radical.

[0079] "Oxo" refers to the =O radical.

[0080] "Isomers" are different compounds with the same molecular formula. "Stereoisomers" are isomers that differ only in the way their atoms are arranged in space, i.e., they have different stereochemical configurations. Enantiomers are a pair of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. The term "(±)" is used to refer to a racemic mixture, where applicable. "Diastereomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. Absolute stereochemistry is specified according to the Cahn-Ingold-Prelog RS system. When a compound is a pure enantiomer, the stereochemistry at each chiral carbon can be specified as either (R) or (S). Resolved compounds of unknown absolute configuration can be designated as (+) or (-) depending on the direction (dextrorotatory or levorotatory) they rotate plane-polarized light at the wavelength of the sodium D line. Some of the compounds described herein contain one or more asymmetric centers and may therefore give rise to enantiomers, diastereomers, and other stereoisomeric forms that can be defined in terms of absolute stereochemistry as (R) or (S). The present chemical compounds, pharmaceutical compositions, and methods are meant to include all such possible isomers, including racemic mixtures, optically pure forms, and intermediate mixtures. Optically active (R)- and (S)-isomers can be prepared using chiral synthons or chiral reagents or resolved using conventional techniques. When a compound described herein contains an olefinic double bond or other center of geometric asymmetry, unless otherwise specified, it is intended that the compound include both the E- and Z-geometric isomers.

[0081] As used herein, "enantiomeric purity" refers to the relative amount, expressed as a percentage, of a particular enantiomer relative to the other enantiomer. For example, if a compound potentially having either (R)- or (S)-isomeric configuration exists as a racemic mixture, the enantiomeric purity is about 50% for either the (R)- or (S)-isomer. If the compound has one isomeric form that is more prevalent than the other (e.g., 80% (S)-isomer and 20% (R)-isomer), the enantiomeric purity of the compound for the (S)-isomeric form is 80%. The enantiomeric purity of a compound can be determined by several methods known in the art, including, but not limited to, chromatography using a chiral support, polarimetry of rotation of polarized light, nuclear magnetic resonance spectroscopy using chiral shift reagents (including, but not limited to, lanthanide-containing chiral complexes or sparkling reagents), or derivatization of the compound with a chiral compound (such as Mosher's acid) followed by chromatography or nuclear magnetic resonance spectroscopy.

[0082] In some embodiments, enantiomerically enriched compositions have higher therapeutic efficacy per unit mass than the racemic mixture of the composition.Enantiomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high-pressure liquid chromatography (HPLC) and chiral salt formation and crystallization, or preferred enantiomers can be prepared by asymmetric synthesis.See, for example, Jacques, et al., Enantiomers, Racemates and Resolutions, Wiley Interscience, New York (1981); E.L. Eliel, Stereochemistry of Carbon Compounds, McGraw-Hill, New York (1962); and E.L. Eliel and S.H. Wilen, Stereochemistry of Organic Compounds, Wiley-Interscience, New York (1994).

[0083] The terms "enantiomerically enriched" and "non-racemic," as used herein, refer to a composition in which the weight percent of one enantiomer is greater than the amount of that enantiomer in a control mixture of racemic composition (e.g., greater than 1:1 weight percent). For example, an enantiomerically enriched preparation of an (S)-enantiomer refers to a preparation of a compound having more than 50% by weight (such as at least 75% by weight, or such as 80% by weight) of the (S)-enantiomer relative to the (R)-enantiomer. In some embodiments, the enrichment can be significantly greater than 80% by weight, providing a "substantially enantiomerically enriched" or "substantially non-racemic" preparation, referring to a preparation of a composition having at least 85% by weight (such as at least 90% by weight, or such as at least 95% by weight) of one enantiomer relative to the other. The terms "enantiomerically pure" or "substantially enantiomerically pure" refer to a composition that contains at least 98% of a single enantiomer and less than 2% of the opposite enantiomer.

[0084] "Moiety" refers to a specific segment or functional group of a molecule. A chemical moiety is often a recognized chemical substance that is embedded in or appended to a molecule.

[0085] "Tautomers" are structurally distinct isomers that interconvert via tautomerization. "Tautomerization" is a form of isomerization and includes prototropic tautomerization or proton shift tautomerization, which are considered a subset of acid-base chemistry. "Prototropic tautomerization" or "proton shift tautomerization" involves the migration of a proton with a change in bond order, often involving the swapping of a single bond with an adjacent double bond. When tautomerization is possible (e.g., in solution), a chemical equilibrium of tautomers can be achieved. An example of tautomerization is keto-enol tautomerization. A specific example of keto-enol tautomerization is the interconversion of the tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one. Another example of tautomerization is phenol-keto tautomerization. A specific example of phenol-keto tautomerization is the interconversion of pyridin-4-ol and pyridin-4(1H)-one tautomers.

[0086] A "leaving group or atom" is any group or atom that cleaves from a starting material under selected reaction conditions, thus facilitating reaction at a specific site. Examples of such groups, unless otherwise specified, include halogen atoms and mesyloxy, p-nitrobenzenesulfonyloxy, and tosyloxy groups.

[0087] "Protecting group" is intended to mean a group that selectively blocks one or more reactive sites in a polyfunctional compound so that a chemical reaction can be selectively carried out at an otherwise unprotected reactive site, and then the group can be easily removed or deprotected after the selective reaction is complete. Various protecting groups are disclosed, for example, in T.H. Greene and P.G.M.Wuts, Protective Groups in Organic Synthesis, 3rd Edition, John Wiley & Sons, New York (1999).

[0088] "Solvate" refers to a compound that is physically associated with one or more molecules of a pharmaceutically acceptable solvent.

[0089] "Substituted" means that the referenced group may have one or more additional groups, radicals, or moieties attached, for example, individually and independently selected from acyl, alkyl, alkylaryl, cycloalkyl, aralkyl, aryl, carbohydrate, carbonate, heteroaryl, heterocycloalkyl, hydroxamate, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, halo, carbonyl, ester, thiocarbonyl, isocyanato, thiocyanato, isothiocyanato, nitro, oxo, perhaloalkyl, perfluoroalkyl, phosphate, silyl, sulfinyl, sulfonyl, sulfonamidyl, sulfoxyl, sulfonate, urea, and amino (including mono- and di-substituted amino groups), and their protected derivatives. The substituents themselves may be substituted; for example, a cycloalkyl substituent may itself have a halide substituent at one or more of its ring carbons. The term "optionally substituted" refers to optional substitution with the specified group, radical, or moiety.

[0090] "Sulfanyl" refers to groups including -S-(optionally substituted alkyl), -S-(optionally substituted aryl), -S-(optionally substituted heteroaryl), and -S-(optionally substituted heterocycloalkyl).

[0091] "Sulfinyl" refers to groups including -S(O)-H, -S(O)-(optionally substituted alkyl), -S(O)-(optionally substituted amino), -S(O)-(optionally substituted aryl), -S(O)-(optionally substituted heteroaryl), and -S(O)-(optionally substituted heterocycloalkyl).

[0092] "Sulfonyl" refers to groups including -S(O2)-H, -S(O2)-(optionally substituted alkyl), -S(O2)-(optionally substituted amino), -S(O2)-(optionally substituted aryl), -S(O2)-(optionally substituted heteroaryl), and -S(O2)-(optionally substituted heterocycloalkyl).

[0093] "Sulfonamidyl" or "sulfonamide" refers to the -S(=O)2-NRR radical, where each R is independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heterocycle (bonded through a ring carbon). The R groups of -NRR of the -S(=O)2-NRR radical, together with the nitrogen to which they are attached, may form a 4-, 5-, 6-, or 7-membered ring. The sulfonamide group is optionally substituted with one or more substituents described for alkyl, cycloalkyl, aryl, and heteroaryl, respectively.

[0094] "Sulfoxyl" refers to the -S(=O)2OH radical.

[0095] "Sulfonate" refers to the radical -S(=O)2-OR, where R is selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heterocycle (bonded through a ring carbon). The sulfonate group is optionally substituted on R with one or more substituents described for alkyl, cycloalkyl, aryl, and heteroaryl, respectively.

[0096] The compounds of the present invention also include crystalline and amorphous forms of the compounds, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, nonsolvated polymorphs (including anhydrous forms), conformational polymorphs, and amorphous forms, and mixtures thereof. "Crystalline form" and "polymorph" are intended to include all crystalline and amorphous forms of the compounds, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, nonsolvated polymorphs (including anhydrous forms), conformational polymorphs, and amorphous forms, and mixtures thereof, unless a specific crystalline or amorphous form is referenced.

[0097] p38 mitogen-activated protein kinase (MAPK), its inhibition, and selective inhibition of p38α The p38 mitogen-activated protein kinase (MAPK) family of stress- and cytokine-activated kinases contributes to the pathogenesis of many human diseases, including cancer, rheumatoid arthritis, cardiovascular disease, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), asthma, acute respiratory distress syndrome (ARDS), and acute lung injury (ALI). Among the many important biological processes regulated by p38 MAPK, regulation of endothelial and epithelial barrier function, leukocyte trafficking, and cytokine expression is central to the pathogenesis of acute and chronic inflammatory disorders. Preclinical studies support pharmacological inhibition of p38 as a promising treatment for inflammatory diseases, but p38 inhibitors have had very limited success in clinical trials due to dose-limiting toxicity and lack of efficacy. Of the 36 Phase II clinical trials of p38 inhibitors listed on www.clinicaltrials.gov, results from only eight studies have been published or listed on ClinicalTrials.gov, demonstrating little clinical benefit and moderate toxicity.

[0098] All available p38 inhibitors block catalytic activity by either directly competing for ATP binding or allosterically inducing a conformational change that prevents ATP from accessing the catalytic site. Davidson et al. identified CMPD1, a purported p38α substrate-selective inhibitor, which selectively inhibited MK2 phosphorylation in in vitro kinase assays but was subsequently shown to bind near the p38α active site and lack substrate selectivity in cells. Nearly all available inhibitors are active against both p38α and p38β, and some are also active against additional isoforms. However, genetic and pharmacological studies have identified p38α as the proinflammatory isoform, while other studies have demonstrated that p38β signaling is cytoprotective. Therefore, inhibition of p38β may contribute to both the lack of efficacy and toxicity of non-isoform-selective p38 inhibitors. However, the broad structural conservation of catalytic modules across most protein kinases presents a challenge in developing catalytic inhibitors with high selectivity, particularly for individual p38 isoforms.

[0099] Without wishing to be bound by any particular theory, even if catalytic inhibitors were absolutely selective for p38α, by design, these compounds would likely block all p38α signaling events, many of which are essential for re-establishing and maintaining homeostasis. For example, p38α not only activates the expression of pro-inflammatory cytokines, but also activates anti-inflammatory cytokines and counter-regulatory dual specificity protein phosphatase-2 (DUSP2) via p38α substrates, MSK1 / 2. The transient decrease and subsequent rebound in serum C-reactive protein (CRP) levels observed in clinical trials of p38 catalytic inhibitors may be caused by the loss of MSK1 / 2-dependent anti-inflammatory signaling.

[0100] As an alternative to catalytic inhibitors, in some embodiments, the compounds and methods of the present invention target a substrate-binding groove in p38α that extends between the two acidic patches, the CD and ED domains, distinct from the DEF substrate-binding pocket. Downstream substrates, upstream activating kinases, and possibly scaffolding molecules all interact with p38 through these sites. In some embodiments, computer-aided drug design (CADD) and / or rational design are used to target low-molecular-weight compounds to a pocket near the p38α ED substrate-binding site, while anti-inflammatory MSK1 / 2 binds to the CD site. The p38α ED substrate-binding site is required for phosphorylation of the p38α substrate MAPK-activated protein kinase-2 (MAPKAPK2, MK2), which is known to mediate endothelial permeability and transendothelial migration (TEM) of neutrophils in vitro and pulmonary edema in a mouse lung injury model. Using this algorithm, p38α-binding compounds with high efficacy were identified. This compound stabilizes endothelial barrier function in human pulmonary microvascular endothelial cells (HMVECLs), inhibits LPS-induced proinflammatory gene expression in THP1 cells, and may be well tolerated and more potent than SB203580 in alleviating experimental ALI.

[0101] Acute lung injury treated with p38α MAPK inhibitors and methods described herein Acute respiratory distress syndrome (ARDS) is a common cause of respiratory failure with a 30-40% mortality rate and a lack of effective treatment options. p38 signaling is crucial for the pathogenesis of ARDS, but clinical trials of p38 inhibitors have been disappointing due to dose-limiting toxicity, despite the fact that all p38 inhibitors inactivate the catalytic site of p38 and block phosphorylation of all p38 substrates.

[0102] Acute respiratory distress syndrome (ARDS) is characterized by the acute onset of non-hydrostatic pulmonary edema, primarily due to neutrophil-mediated damage to the alveolar epithelium and capillary endothelial barrier dysfunction. The complex network of pro- and anti-inflammatory mediators activated during ARDS is crucial for the pathogenesis of acute lung injury (ALI) and the multiple organ failure that often accompanies ARDS. However, therapeutic agents targeting pro-inflammatory mediators have proven ineffective in ARDS. Lung parenchymal damage leads to reduced compliance, intrapulmonary shunting, and ventilation-perfusion mismatch, usually resulting in the need for mechanical ventilation. However, the cyclic recruitment / derecruitment and hyperinflation of alveoli caused by mechanical ventilation itself can cause neutrophil-dependent inflammation and lung injury, even in previously normal lungs. Understanding this mechanism has led to a phase III randomized clinical trial demonstrating that low-tidal-volume mechanical ventilation improves survival in patients with ARDS. Two additional supportive strategies have been shown to improve mortality in patients with severe ARDS, neuromuscular blockade and prone positioning. A third intervention, restrictive fluid management, has been shown to reduce time on mechanical ventilation and length of ICU stay but did not reduce mortality. Despite these improvements in supportive care, mortality in patients with ARDS remains 30-40% in the United States, representing approximately 74,500 deaths per year, highlighting the importance of developing new therapeutic agents that target relevant pathogenic mechanisms.

[0103] p38 mitogen-activated protein kinases (MAPKs) are a family of stress- and cytokine-activated kinases activated by many of the pathogenic signals associated with ARDS, including inflammatory mediators, thermal hyperthermia (FRH), and cyclic stretching. p38 MAPKs are activated in patients at risk for ARDS, and as discussed below, p38 MAPKs are involved in multiple processes that contribute to the pathogenesis of ARDS, making this MAPK family an attractive therapeutic target. As proof of concept, SB203580, a prototypic pyridinylimidazole compound, has been shown to inhibit the kinase activity of p38α and p38β, but not p38γ or p38δ, blocking multiple processes that contribute to the pathogenesis of ARDS. Endothelial p38 signaling is activated by neutrophil binding and is required for neutrophil transendothelial migration (TEM). Cross-linking of ICAM-1 on human umbilical vein endothelial cells (HUVECs) stimulates p38α activation, HSP27 phosphorylation, F-actin reorganization, ICAM-1 aggregation, and cell stiffening, and increases neutrophil migration to HUVEC intercellular junctions, all of which are blocked by SB203580. Cross-linking of E-selectin on HUVECs activates p38 and p38-dependent cytoskeletal reorganization, stress fiber formation, and neutrophil TEM. Similarly, ligation of ICAM-1 to β2 integrin on neutrophils activates neutrophil p38 and p38-dependent chemokinesis and chemotaxis. Pretreatment with a p38 inhibitor was protective in mouse models of lung injury associated with ventilator-induced lung injury, complement-induced lung injury, and the intestinal perforation model of sepsis, but not against hemorrhage- and endotoxemia-induced lung injury. The excessive impairment of endothelial barrier function caused by FRH (a 2–3°C increase in core temperature) in experimental ALI was shown to be associated with p38 activation and blocked by SB203580. The efficacy of SB203580 in mitigating acute lung injury and multiple pathogenic processes contributing to relatively high p38α and p38β expression in human lungs supports the central role of these two p38 isoforms in the development of ARDS.

[0104] Despite these compelling preclinical data, only one clinical trial has attempted to evaluate p38 inhibition as a treatment strategy in ARDS. This early Phase IIa trial of SB-681323 / dilmapimod in patients at risk for ARDS (ClinicalTrials.gov number NCT00996840) showed that dilmapimod was safe at the administered dose and moderately reduced serum C-reactive protein (CRP) levels, but was not powered to analyze its effect on the incidence or severity of ARDS. Currently, www.clinicaltrials.gov lists a total of 74 clinical trials of p38 inhibitors, including 26 Phase I, 47 Phase II, and one Phase III trial. Phase II and III trials have examined the safety and efficacy of 10 different p38 catalytic inhibitors for 13 different diseases / indications, including analgesia (6 trials), osteoarthritis (2 trials), rheumatoid arthritis (13 trials), Alzheimer's disease (2 trials), ankylosing spondylitis (1 trial), cardiomyopathy (1 trial), psoriasis (2 trials), atherosclerosis (5 trials), depression (2 trials), COPD (8 trials), ARDS risk (1 trial), cancer (4 trials), and glomerulosclerosis (1 trial). While only a portion of the data is in the public domain, the failure of most of these drugs appears to be due to their side effect profile or lack of efficacy at the doses used. Of the 48 Phase II and III trials, 36 have been completed and 3 have been terminated prematurely, but the results of only 8 trials have been published or listed on ClinicalTrials.gov. Two trials of VX-702 in rheumatoid arthritis showed a small increase in the proportion of treated subjects with ACR20 symptom scores compared to placebo. Of two published studies of p38 inhibitors for pain, one reported a mild reduction in pain, and the other showed no effect. Of two published studies of p38 inhibitors in COPD, one showed no effect, and the other showed an increase in FEV1 / 100 ml and a decrease in serum CRP levels in the treatment group, but with associated toxicities (rash, pharyngitis, QTc prolongation). GW85655 (rosmapimod) improved vasorelaxation and reduced serum CRP in patients with hypercholesterolemia.In a ninth clinical trial not listed on clinicaltrials.gov, BIRB796 (doramapimod) transiently reduced serum CRP levels in patients with Crohn's disease without clinical benefit. Collectively, these studies demonstrate the therapeutic potential of p38 inhibition in a wide range of human diseases but emphasize the limited efficacy of currently available p38 inhibitors at doses that can be safely administered to humans.

[0105] p38 MAPK, like most protein kinases, shares a conserved bilobed structure and catalytic site, with its hydrophobic ATP-binding pocket located between the N- and C-terminal lobes. While most available protein kinase inhibitors compete with ATP for binding to the ATP-binding pocket of the catalytic site, the extensive structural conservation of the catalytic module across most protein kinases presents a challenge for developing highly specific catalytic p38 inhibitors. The pyridinylimidazole inhibitor SB203580 binds to the ATP-binding site of p38α and p38β, but its access to the ATP-binding site of p38γ and p38δ is blocked by a bulky methionine. However, proteomic analysis revealed sub-μM IC 50 Several additional kinases inhibited by SB203580 were identified in , including Rip-like interacting caspase-like apoptosis-regulating protein kinase (RICK / Rip2), casein kinase (CK)-1δ, and cyclin G-associated kinase (GAK).

[0106] High-throughput biochemical screening for p38 inhibitors has uncovered a new class of diaryl urea compounds. Rather than directly binding to the ATP-binding pocket, these compounds bind to an allosteric site that induces a conformational change in p38 that prevents ATP access to the catalytic ATP-binding pocket. Three allosteric p38 inhibitors, BIRB796 / dolmapimod, GW856553 / rosmapimod, and SB-681323 / dilmapimod, have entered clinical trials but, like ATP-competitive inhibitors, have not progressed beyond Phase II, with the exception of the LATITUDE trial and the ongoing Phase III trial of losmapimod in patients with acute coronary syndromes (clinicaltrials.gov, number NCT02145468). Because allosteric inhibitors are not affected by the presence of the gatekeeper methionine, these compounds inhibit all four p38 isoforms. However, BIRB796 also exhibits an IC of 0.1 μM. 50 Jnk2α2 at 77°C, and an IC of 1.4 μM 50 The lack of specificity of ATP-competitive and allosteric inhibitors of p38 is likely a major cause of off-target toxicity.

[0107] An equally important source of toxicity for p38 inhibitors likely stems from the broad range of functions of each p38 MAPK isoform. Both types of inhibitors block the catalytic site of p38, and ATP-competitive and allosteric inhibitors block all p38 phosphorylation events. Because p38 phosphorylates at least 66 known substrates with important biological activities, dose-limiting toxicity may be unavoidable with these agents.

[0108] MAPK p38 and ERK family members share a structural feature: a substrate-binding groove located in the C-terminal lobe of the protein opposite the catalytic domain. The binding groove extends between two acidic patches, the CD and ED domains. This region of p38 not only binds p38 substrates but also upstream kinases and scaffolding proteins. Our group previously used computer-aided drug design (CADD) to identify small molecules that target the substrate-binding groove rather than the catalytic module of ERK2, thereby developing a new class of ERK1 / 2 MAPK inhibitors with improved toxicity profiles. As described herein, a similar strategy can be used to identify small molecular weight compounds that target a pocket near the ED substrate-binding site of p38α, which is required for phosphorylation of MK2, a p38 substrate known to mediate pulmonary endothelial permeability and pulmonary edema in vitro in a mouse lung injury model. CADD was used to search a database of commercially available compounds and identified 150 small molecular weight compounds predicted to bind to the target pocket near the ED binding site of the p38α target. Twenty structurally distinct compounds were obtained from this list and screened for selective binding to p38α but not ERK2 by differential scanning fluorometry (DSF), and then analyzed for their ability to reduce pathogenic endothelial barrier alterations in human pulmonary microvascular endothelial cells (HMVECL), reduce cytokine expression in THP1 monocytes in vitro, and alleviate ALI induced in mice. Of the 20 CADD-selected compounds tested, five bound to p38α with sufficient affinity to be detected by DSF, two selectively bound to p38α but not ERK2, and they were more effective than SB203580 in stabilizing endothelial barrier function in vitro. One of these compounds was well tolerated and more potent than SB203580 in alleviating experimental ALI.

[0109] In certain embodiments, the p38α MAPK inhibitors described herein may be used in the treatment of acute respiratory distress syndrome (ARDS) and / or acute lung injury (ALI).

[0110] p38α MAPK inhibitors and methods for inhibiting p38α MAPK In one embodiment, the present invention includes compounds that can be p38α MAPK inhibitors and / or modulators of p38α MAPK protein activity, for example, compounds that can bind to a pocket near the ED substrate docking site of p38α MAPK, or pharmaceutically acceptable salts, solvates, hydrates, cocrystals, or prodrugs thereof. In one embodiment, the p38α MAPK inhibitor is a selective inhibitor of p38α MAPK. In one embodiment, the p38α MAPK inhibitor binds to p38α MAPK near the substrate binding groove of p38α MAPK that extends between the two acidic patches, the CD domain and the ED domain. In another embodiment, the p38α MAPK inhibitor causes inhibition of MK2 phosphorylation.

[0111] In one embodiment, lead compounds for p38α MAPK inhibitors have been identified that have favorable biological effects in human cell culture models and mouse models of inflammatory lung injury. In one embodiment, the p38α MAPK inhibitors have been identified using a CADD strategy. The CADD target pocket of p38α differs from the corresponding pocket of p38β by three out of ten amino acids, providing an opportunity for p38α selectivity. In some embodiments, the sequence of the target pocket includes at least amino acids R49, H107, L108, and K165 of p38α MAPK. In some embodiments, the sequence of the target pocket is R49, H107, L108, M109, G110, A157, V158, E163, L164, and K165 of p38α MAPK. In one embodiment, the p38α MAPK inhibitors have been identified using a rational design strategy.

[0112] In some embodiments, the p38α MAPK inhibitor is a compound of formula A, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof: [ka] In Formula A, Ar and Ar 1is independently selected from monocyclic or polycyclic optionally substituted cycloalkyl, monocyclic or polycyclic optionally substituted heterocycloalkyl, monocyclic or polycyclic optionally substituted aryl, monocyclic or polycyclic optionally substituted arylalkyl, monocyclic or polycyclic optionally substituted heteroaryl, and monocyclic or polycyclic optionally substituted heteroarylalkyl; R 1 and R 2 are independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted alkylheteroaryl; R 1 and R 2 can be optionally linked to form a carbocyclic or heterocyclic ring, and L 1 and L 2 are independently a bond, -NR a -, -S-, -S(O)-, -S(O)2-, -O-, -CR a 2-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NR a -, -NR a C(O)-, -C(O)NR a SO2-, -SO2NR a C(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NR a -, -NR a C(O)O-, -S(O) t N(R a )-(t is 1 or 2), -N(R a )S(O) t-(t is 1 or 2), disubstituted alkyl, disubstituted heteroalkyl, disubstituted alkenyl, disubstituted alkynyl, disubstituted cycloalkyl, disubstituted heterocycloalkyl, disubstituted aryl, disubstituted arylalkyl, disubstituted heteroaryl, and disubstituted heteroarylalkyl, any optional substituent independently at each occurrence is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -OR a , -SR a , -OC(O)-R a , -SC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a , -C(O)SR a , -OC(O)N(R a )2, -C(O)N(R a )2, -N(R a )C(O)OR a , -N(R a )C(O)R a , -N(R a )C(O)N(R a )2, -N(R a )C(NR a )N(R a )2, -N(R a )S(O) t R a (t is 1 or 2), -S(O) t R a (t is 1 or 2), -S(O) t OR a (t is 1 or 2), -S(O) t N(R a )2 (t is 1 or 2), and PO3 (Ra )2 and R a is independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl. In some embodiments, the optional substituents are independently selected from R 3 , R 4 , R 5 , and R 6 are selected from, independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted alkylheteroaryl; R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 Any two of Ar can be optionally linked to form a carbocyclic or heterocyclic ring. 1 is an optionally substituted aryl ring or an optionally substituted heteroaryl ring. In some embodiments, Ar is a monocyclic or polycyclic optionally substituted aryl, a monocyclic or polycyclic optionally substituted arylalkyl, a monocyclic or polycyclic optionally substituted heteroaryl, or a monocyclic or polycyclic optionally substituted heteroarylalkyl. In some embodiments, Ar is a 5- or 6-membered optionally substituted aryl or a 5- or 6-membered optionally substituted heteroaryl, and L 1 and L 2is linked to Ar in a 1,2, 1,3, or 1,4 substitution pattern. In some embodiments, Ar is a 1,2-, 1,3-, or 1,4-disubstituted phenyl, pyridine, pyrimidine, pyrazine, or triazine. In some embodiments, Ar is a 1,2-, or 1,3-disubstituted furan, thiophene, pyrrole, thiazole, imidazole, oxazole, triazole, or pyrazole. In some embodiments, Ar is a 1,2-, 1,3-, 1,4-, 1,5-, 1,6-, 1,7-, or 1,8-disubstituted naphthalene, quinoline, isoquinoline, or quinazoline. In some embodiments, Ar is a 1,2-, 1,3-, 1,4-, 1,5-, 1,6-, or 1,7-disubstituted indole or imidazole. In some embodiments, L 1 is a linker selected from -CH2-, -C(CH3)2-, and -C(CH2CH2)-; L 2 is a linker selected from -NHCH2-, -CH2NH-, -NHCO-, -CONH-, -SON2NH-, and -NHSO2-, and -NR 1 R 2 teeth, [ka] Selected from R 7 is hydrogen or optionally substituted alkyl, and Ar 2 is a heterocycle.

[0113] In one embodiment, the p38α MAPK inhibitor is a compound of Formula I, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof: [ka]

[0114] In formula I, R 1 , R 2 , R 3 , R 4 , R 5 , and R6 are each independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted alkylheteroaryl; and R 1 and R 2 can be optionally linked to form a carbocyclic or heterocyclic ring, and L 1 is a linker selected from -CH2-, -C(CH3)2-, and -C(CH2CH2)-; L 2 is a linker selected from -NHCH2-, -CH2NH-, -NHCO-, -CONH-, -SO2NH-, and -NHSO2-; Ar 1 is an optionally substituted aryl ring or an optionally substituted heteroaryl ring. In some embodiments, -NR of Formula I 1 R 2 The group is selected from: [ka] In the formula, R 7 is hydrogen or optionally substituted alkyl, and Ar 2 is a heterocycle. In some embodiments, —NR 1 R 2 The group is selected from the following: [ka] In some embodiments, L 1 is -CH-. In some embodiments, R 3 , R 4 , R 5 , and R 6 is hydrogen. In some embodiments, L 2 is selected from -NHCH2-, -NHCO-, and -NHSO2-. In some embodiments, Ar 1 is selected from: [ka] In the formula, R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , and R 13 each independently represents hydrogen, halogen, -NR 1 R 2 , alkoxy, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkoxy, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted alkylheteroaryl; R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , and R 13 Any two vicinal rings of Ar can be optionally linked to form a carbocyclic or heterocyclic ring. 1 is selected from: [ka]

[0115] In one embodiment, the p38α MAPK inhibitor is a compound of formula II, which may be selected from the compounds of any one of formulas 1001-1180 in Table 1. [ka] [Table 2-1] [Table 2-2] [Table 2-3] Table 2-4 Table 2-5 Table 2-6 Table 2-7 Table 2-8 Table 2-9 Table 2-10 Table 2-11 Table 2-12 Table 2-13 Table 2-14 Table 2-15 Table 2-16 Table 2-17 [Table 2-18] [Table 2-19] [Table 2-20] [Table 2-21]

[0116] In one embodiment, the p38α MAPK inhibitor is a compound of any of formulas 1001 (SF-6-221), 1032 (SF-7-008), SF-7-009, 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), and 1087 (SF-7-044). [Table 3-1] [Table 3-2]

[0117] Using complementary techniques, we confirmed the selective binding of UM101 to p38α. DSF detected ligand-induced protein stabilization, showing that UM101 caused a concentration-dependent increase in the melting temperature of p38α, but not p38β (Figure 3d). Compared to SB203580, the smaller effect of UM101 on p38α melting suggests a lower p38α binding affinity for substrate-selective inhibitors, similar to that of catalytic inhibitors of substrate-selective ERK inhibitors. The smaller effect of SB203580 on p38β compared to p38α is consistent with its known affinity for p38α, approximately 10-fold higher. STD-NMR, which measures low-affinity protein:ligand binding via nonscalar magnetization transfer from protein to ligand protons, confirmed specific UM101 binding to p38α, with the interaction localized to its aromatic ring. Furthermore, the binding of UM101 to its CADD target was confirmed by showing that mutating four of the ten amino acids in the target pocket abolished UM101 binding while maintaining SB203580 binding.

[0118] In some embodiments, the p38α MAPK inhibitor causes a concentration-dependent increase in the melting temperature of p38α MAPK. The difference in melting temperature ΔTm (°C) is measured at a p38α MAPK inhibitor concentration of 1 nM to 1000 μM. In one embodiment, the difference in melting temperature ΔTm (°C) is measured at a p38α MAPK inhibitor concentration of 100 μM. In one embodiment, ΔTm is about 0.1 to about 2°C. In one embodiment, ΔTm is about 0.01 to about 0.05°C. In one embodiment, ΔTm is about 0.01 to about 0.1°C. In one embodiment, ΔTm is about 0.03 to about 0.7°C. In one embodiment, ΔTm is about 0.06 to about 1.5°C. In one embodiment, ΔTm is about 1°C to about 2°C. In one embodiment, ΔTm is about 1.5 to about 2°C. In one embodiment, ΔTm is about 0.1°C. In one embodiment, ΔTm is about 0.2°C. In one embodiment, ΔTm is about 0.3°C. In one embodiment, ΔTm is about 0.4°C. In one embodiment, ΔTm is about 0.5°C. In one embodiment, ΔTm is about 0.6°C. In one embodiment, ΔTm is about 0.7°C. In one embodiment, ΔTm is about 0.8°C. In one embodiment, ΔTm is about 0.9°C. In one embodiment, ΔTm is about 1°C. In one embodiment, ΔTm is about 1.1°C. In one embodiment, ΔTm is about 1.2°C. In one embodiment, ΔTm is about 1.3°C. In one embodiment, ΔTm is about 1.4°C. In one embodiment, ΔTm is about 1.5°C. In one embodiment, ΔTm is about 1.6°C. In one embodiment, ΔTm is about 1.7°C. In one embodiment, ΔTm is about 1.8°C. In one embodiment, ΔTm is about 1.9°C. In one embodiment, the ΔTm is about 2°C. In one embodiment, the ΔTm is about 0.735°C. In one embodiment, the ΔTm is about 0.667°C.

[0119] In some embodiments, the p38α MAPK inhibitor has a log P of about -5 to about 10. In some embodiments, the p38α MAPK inhibitor has a log P of about -3 to about 8. In some embodiments, the p38α MAPK inhibitor has a log P of about 0 to about 5. In some embodiments, the p38α MAPK inhibitor has a log P of about 0.1 to about 3. Log P is a measure of drug solubility and is defined as the logarithm of the octanol / water partition coefficient of the drug. In one embodiment, the p38α MAPK inhibitor has a log P of about 0.1 to about 1. In one embodiment, the p38α MAPK inhibitor has a log P of about 0.5 to about 1.5. In one embodiment, the p38α MAPK inhibitor has a log P of about 0.75 to about 2. In one embodiment, the p38α MAPK inhibitor has a log P of about 1 to about 2.5. In one embodiment, the p38α MAPK inhibitor has a log P of about 1.75 to about 3. In one embodiment, the p38α MAPK inhibitor has a log P of about 0.1. In one embodiment, the p38α MAPK inhibitor has a log P of about 0.25. In one embodiment, the p38α MAPK inhibitor has a log P of about 0.5. In one embodiment, the p38α MAPK inhibitor has a log P of about 0.75. In one embodiment, the p38α MAPK inhibitor has a log P of about 1. In one embodiment, the p38α MAPK inhibitor has a log P of about 1.25. In one embodiment, the p38α MAPK inhibitor has a log P of about 1.5. In one embodiment, the p38α MAPK inhibitor has a log P of about 1.75. In one embodiment, the p38α MAPK inhibitor has a log P of about 2. In one embodiment, the p38α MAPK inhibitor has a log P of about 2.25. In one embodiment, the p38α MAPK inhibitor has a log P of about 2.5. In one embodiment, the p38α MAPK inhibitor has a log P of about 2.75. In one embodiment, the p38α MAPK inhibitor has a log P of about 3. In one embodiment, the p38α MAPK inhibitor has a log P of about 0.28. In one embodiment, the p38α MAPK inhibitor has a log P of about 2.31.

[0120] Phosphorylation of MK2 requires binding to the ED site adjacent to the CADD target pocket in p38α MAPK. In some embodiments, the target pocket is defined by at least amino acids R49, H107, L108, and K165 of p38α MAPK. In some embodiments, the target pocket is defined by amino acids selected from the group consisting of R49, H107, L108, M109, G110, A157, V158, E163, L164, and K165 of p38α MAPK, and combinations thereof. In some embodiments, the target pocket is defined by amino acids R49, H107, L108, M109, G110, A157, V158, E163, L164, and K165 of p38α MAPK. Western blot analysis confirmed that UM101 partially inhibited MK2 phosphorylation in anisomycin-stimulated HeLa cells, but this was less than that of 10 μM SB203580, which had IC values ​​of 0.01 for p38α and p38β, respectively. 50 Concentrations 200- and 20-fold higher than MK2 failed to completely block MK2 phosphorylation in HeLa cells, which may reflect contributions from p38γ or p38δ, as both isoforms are expressed.

[0121] In one embodiment, the present invention relates to a method for inhibiting p38α MAPK, whereby inhibition of p38α MAPK stabilizes endothelial or epithelial barrier function. The selective p38α-binding compounds UM60 and UM101 both exhibited endothelial barrier stabilization and macrophage cytokine modulation effects similar to those of SB203580, validating the ED targeting strategy. UM101 stabilized the endothelial barrier more effectively than SB203580 (Figures 2a and 2b), despite having less effect on MK2 phosphorylation. In one embodiment, endothelial barrier permeability can be measured by exposure to TNFα and hyperthermia, either separately or in combination, followed by measurement of permeability to 10 kDa dextran. In one embodiment, endothelial barrier stabilization is assessed by measuring permeability after pretreatment with a compound of the present invention, and stabilization is expressed as a percentage reduction in the increase in permeability before and after pretreatment. Pretreatment with a p38α MAPK inhibitor can be performed at various concentrations, for example, 10, 25, 50, or 100 μM. In one embodiment, the increase in permeability to 10 kDa dextran can be reduced by 5% to more than 100%. In one embodiment, the increase in permeability is reduced by about 5%. In one embodiment, the increase in permeability is reduced by about 10%. In one embodiment, the increase in permeability is reduced by about 20%. In one embodiment, the increase in permeability is reduced by about 30%. In one embodiment, the increase in permeability is reduced by about 40%. In one embodiment, the increase in permeability is reduced by about 50%. In one embodiment, the increase in permeability is reduced by about 60%. In one embodiment, the increase in permeability is reduced by about 70%. In one embodiment, the increase in permeability is reduced by about 80%. In one embodiment, the increase in permeability is reduced by about 90%. In one embodiment, the increase in permeability is reduced by about 100%. In one embodiment, the increase in permeability is reduced by more than 100%. In one embodiment, the increase in permeability is reduced by about 71%. In one embodiment, the increase in permeability is reduced by about 74%. In one embodiment, the increase in permeability is reduced by about 89%. In one embodiment, the increase in permeability is reduced by about 100%.

[0122] Because UM101 stabilized the endothelial barrier more effectively than SB203580 (Figures 2a and 2b) despite having less of an effect on MK2 phosphorylation (Figure 3c), we assessed additional molecular effects by comparing the effects of UM101 and SB203580 on global gene expression using RNA-Seq in TNFα-treated HMVECLs. TNFα increased the expression of 511 genes by more than twofold, of which 61 were reduced, and pretreatment with 10 μM SB203580 increased 38. Despite using a UM101 concentration greater than 10-fold that required to stabilize barrier function in HMVECLs (Figures 2a and 2b), UM101 only modified the expression of 38 of the 99 SB203580-modified genes. PathwayNet analysis showed that UM101 blocked only 7 of the 15 SB203580-blocked transcription factors. MSK1 / 2 were among those spared by UM101, which, given the anti-inflammatory effects of MSK1 / 2, is advantageously consistent with the targeting strategy of UM101 to the ED site.

[0123] The partial functional overlap between UM101 and SB203580 revealed by RNA-Seq is consistent with the design of UM101 as a selective inhibitor of non-catalytic substrates, but may also be the result of off-target effects of SB203580, including receptor-interacting protein kinase-2, cyclin G-associated kinase, and casein kinase-1δ. However, none of the SB203580-inhibited transcription factors identified by PathwayNet analysis are known substrates of these kinases as analyzed using PhosphoNetworks.

[0124] Although the high concentrations of UM101 used in this analysis may have caused some p38-independent effects, the data presented here support the conclusion that UM101 exerts its biological effects primarily by modifying p38α: (1) DSF and STD-NMR demonstrate p38α-specific binding of UM101, (2) UM101 p38α binding is abolished by mutating four of the ten target pocket amino acids, (3) both UM60 and UM101 bind to p38α and affect endothelial function similarly to SB203580, (4) UM101 partially blocked phosphorylation of the p38 substrates MK2 and Stat-1 in TNFα-stimulated HeLa cells, and (5) UM101 inhibited the expression of approximately half of the genes inhibited by SB203580. UM101 may be more effective than SB203580 in stabilizing the endothelial barrier. This is due to the selective sparing of potential counter-regulatory genes, such as GM-CSF, MSK1 / 2-dependent anti-inflammatory genes, and p38β-dependent pro-survival genes.

[0125] In one embodiment, the present invention relates to a method for inhibiting p38α MAPK, comprising contacting p38α MAPK with a compound capable of binding to a pocket near the ED substrate docking site of p38α MAPK, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof. In one embodiment, the compound selectively inhibits p38α MAPK. In one embodiment, the p38α MAPK inhibitor binds to p38α MAPK near the substrate-binding groove of p38α MAPK, which extends between two acidic patches, the CD domain and the ED domain. In one embodiment, the binding pocket is defined by at least residues R49, H107, L108, and K165 of p38α MAPK. In one embodiment, the binding pocket is defined by residues R49, H107, L108, M109, G110, A157, V158, E163, L164, and K165 of p38α MAPK. In some embodiments, the p38α MAPK inhibitor causes a concentration-dependent increase in the melting temperature of p38α MAPK, hi other embodiments, the p38α MAPK inhibitor causes inhibition of MK2 phosphorylation. In one embodiment, the compound is any compound of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly any compound of Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof.

[0126] In one embodiment, the present invention relates to a method for inhibiting p38α MAPK, wherein inhibition of p38α MAPK does not result in a loss of p38α-dependent counterregulatory responses. In some embodiments, the p38α-dependent counterregulatory responses involve mitogen- and stress-activated protein kinase-1 (MSK1) or MSK2. By targeting a pocket near the ED substrate docking site of p38α, the inhibitors described herein avoid interference with CD4+-specific substrates, including MSK1 / 2, thereby limiting inflammation through the expression of IL-10 and DUSP2. Among the effects of MSK1 / 2 deletion in mice is increased and prolonged LPS-induced expression of IL-6, a CRP regulator, suggesting a possible mechanism for the rebound in serum CRP observed in some clinical trials of catalytic p38 inhibitors.

[0127] In one embodiment, the present invention relates to a method for inhibiting p38α MAPK, whereby inflammation is reduced. In one embodiment, the effect of a p38α MAPK inhibitor on inflammatory cytokine expression is compared by pretreating PMA-differentiated THP1 cells with a p38α MAPK inhibitor, then stimulating them with LPS, collecting RNA after a period of time, and analyzing it using a PCR-based cytokine array. In some embodiments, the p38α MAPK inhibitor inhibits the expression of various genes, such as IL-1A, IL-8, TNFSF8, CXCL5, CCL7, CCL17, TNFSF9, IL-1B, CXCL1, TNFSF15, CCL5, CCL4, CCL20, CXCL2, TNF, or BMP6. In some embodiments, the p38α MAPK inhibitor inhibits the expression of Smad3, which drives the differentiation of Foxp3 T regulatory cells and suppresses interferon-γ. The p38α MAPK inhibitor can be used at any suitable concentration (e.g., 10, 25, 50, or 100 μM). In one embodiment, the reduction in inflammation is measured by comparing the fold change in mRNA levels at various concentrations of the p38α MAPK inhibitor with unstimulated PMA-differentiated THP1 cells.

[0128] In some embodiments, p38α MAPK inhibitors regulate TNFα-induced gene expression in HMVECLs, as demonstrated by RNA Seq. In one embodiment, HMVECLs are pretreated with an appropriate concentration (for example, 10 μM or 100 μM) of p38α MAPK inhibitor for a certain period of time, and then stimulated with TNFα for a certain period of time. The p38α MAPK inhibitors of the present invention inhibit genes such as PRRG4, TSLP, CCL17, EXOC3L4, MMP9, IDO1, CXCL10, CD200, SLC15A3, VDR, IL1B, GPR88, CD207, TCHH, HAS3, GBP1P1, MUC4, ELOVL7, CXCL11, GBP4, PLA1A, or CXCL5.

[0129] In one embodiment, the present invention relates to a method for inhibiting p38α MAPK, thereby reducing lung injury (including, but not limited to, LPS-induced lung injury) in a subject. In one embodiment, the efficacy of p38α MAPK inhibitors in reducing transalveolar protein and neutrophil extravasation was compared in a mouse model of LPS / hyperthermia-induced ALI (Figures 2c and 2d). In one embodiment, subjects receive intraperitoneal injection(s) of a p38α MAPK inhibitor at concentrations of 100, 250, 300, 400, 500, 750, 1000 μg, etc. in a suitable carrier (e.g., DMSO) for a period of time prior to intratracheal instillation of LPS and / or transfer to a heating chamber. Lung lavage fluid from the subjects is measured for protein and / or neutrophils. Compared to control subjects, subjects pretreated with a p38α MAPK inhibitor have reduced lavage protein concentrations and neutrophil content. In some embodiments, the reduction is between about 5% and about 100%. In one embodiment, the reduction is greater than about 5%. In one embodiment, the reduction is greater than about 10%. In one embodiment, the reduction is greater than about 20%. In one embodiment, the reduction is greater than about 30%. In one embodiment, the reduction is greater than about 40%. In one embodiment, the reduction is greater than about 50%. In one embodiment, the reduction is greater than about 60%. In one embodiment, the reduction is greater than about 70%. In one embodiment, the reduction is greater than about 80%. In one embodiment, the reduction is greater than about 90%. In one embodiment, the reduction is about 100%. In one embodiment, the reduction is less than about 10%. In one embodiment, the reduction is less than about 20%. In one embodiment, the reduction is less than about 30%. In one embodiment, the reduction is less than about 40%. In one embodiment, the reduction is less than about 50%. In one embodiment, the reduction is less than about 60%. In one embodiment, the reduction is less than about 70%. In one embodiment, the reduction is less than about 80%. In one embodiment, the reduction is less than about 90%. In one embodiment, the reduction is about 100%. In one embodiment, the reduction is about 44.1%. In one embodiment, the reduction is about 43.9%. In one embodiment, the reduction is about 92.9%. In one embodiment, the reduction is about 44.4%. In one embodiment, the reduction is about 49.5%.In one embodiment, the reduction is about 55.3%. In one embodiment, the reduction is about 54%.

[0130] In one embodiment, the present invention relates to a method of inhibiting p38α MAPK, thereby modulating leukocyte trafficking.

[0131] In one embodiment, the present invention relates to a method of inhibiting p38α MAPK, thereby modulating cytokine expression.

[0132] Treatment method The compounds and compositions described herein can be used in methods for treating diseases.In some embodiments, the compounds and compositions described herein can be used in methods for treating diseases associated with upregulation and / or downregulation of p38α MAPK protein.

[0133] In one embodiment, the present invention relates to a method of treating a disease alleviated by inhibiting p38α MAPK protein in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a p38α MAPK inhibitor, wherein the p38α MAPK inhibitor is a compound capable of binding to a pocket near the ED substrate docking site of p38α MAPK, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof. In one embodiment, the binding pocket is defined by at least residues R49, H107, L108, and K165 of p38α MAPK. In one embodiment, the binding pocket is defined by residues R49, H107, L108, M109, G110, A157, V158, E163, L164, and K165 of p38α MAPK. In one embodiment, the p38α MAPK inhibitor is a compound of any of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly any of Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof. In some embodiments, the p38α MAPK inhibitor is a selective inhibitor of p38α MAPK.

[0134] In one embodiment, the present invention relates to a method of treating a disease alleviated by inhibiting p38α MAPK protein in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a p38α MAPK inhibitor in dosage unit form, which in one embodiment comprises a physiologically compatible carrier medium.

[0135] In one embodiment, the present invention relates to a method of treating a disease that can be alleviated by inhibiting p38α MAPK protein in a patient in need thereof, comprising administering a therapeutically effective amount of a p38α MAPK inhibitor to the patient, wherein the disease is cancer or an inflammatory disease. In some embodiments, the disease is rheumatoid arthritis, cardiovascular disease, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), asthma, acute respiratory distress syndrome (ARDS), or acute lung injury (ALI). In one embodiment, the disease is a hyperproliferative disease. In some embodiments, the hyperproliferative disorder is cancer. In some embodiments, the cancer is selected from the group consisting of pancreatic cancer, breast cancer, prostate cancer, lymphoma, skin cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain cancer, head-neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck carcinoma, breast cancer, ovarian carcinoma, lung cancer, small cell lung cancer, Wilms' tumor, cervical cancer, testicular cancer, bladder carcinoma, pancreatic carcinoma, gastric cancer, colon carcinoma, prostate cancer carcinoma), genitourinary cancer, thyroid cancer, esophageal cancer, myeloma, multiple myeloma, adrenal cancer, renal cell carcinoma, endometrial cancer, adrenocortical carcinoma, malignant pancreatic insulinoma, malignant carcinoid cancer, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic granulocytic leukemia, acute granulocytic leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, polycythemia vera, essential thrombocytosis, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, osteosarcoma, primary macroglobulinemia, or retinoblastoma.In other embodiments, the cancer is acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, cholangiocarcinoma, bladder cancer, brain cancer, breast cancer, bronchogenic carcinoma, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngeal carcinoma, cystadenocarcinoma, embryonal carcinoma, endothelioma, ependymoma, epithelial carcinoma, esophageal carcinoma, Ewing's tumor, fibrosarcoma, gastric cancer, glioblastoma multiforme, glioma, head and neck cancer. Cancer, hemangioblastoma, hepatocellular carcinoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendothelial sarcoma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteogenic sarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma tumor, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, gastric cancer, sweat gland carcinoma, synovium, testicular cancer, small cell lung cancer, pharyngeal cancer, uterine cancer, Wilms' tumor, blood cancer, acute erythroleukemia, acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monoblastic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute nonlymphocytic leukemia, acute promyelocytic leukemia, acute anaplastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin's lymphoma, polycythemia vera, or Waldenstrom's macroglobulinemia.

[0136] In some embodiments, the hyperproliferative disorders (eg, cancer) treated by the compounds and compositions described herein include cells that have expression of p38α MAPK protein and / or p38α MAPK-related proteins.

[0137] In one embodiment, the present invention relates to a method of treating a disease that is alleviated by inhibiting p38α MAPK protein in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a p38α MAPK inhibitor, wherein the p38α MAPK inhibitor is a compound capable of binding to a pocket near the ED substrate docking site of p38α MAPK, or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof, and one or more additional therapeutic agents (including chemotherapeutic and / or immunotherapeutic agents).

[0138] The effectiveness of the compounds and compound combinations described herein in treating the indicated diseases or disorders can be tested using various models known in the art and described herein, which provide guidance for the treatment of human diseases. Any and all of the described treatment methods can include medical follow-up to determine the therapeutic or prophylactic effect achieved in subjects treated with the compound(s) and / or composition(s) described herein.

[0139] Pharmaceutical Composition In one embodiment, the active pharmaceutical ingredient or combination of active pharmaceutical ingredients (eg, any of the p38α MAPK inhibitors of the present invention) is provided as a pharmaceutically acceptable composition.

[0140] In one embodiment, the present invention relates to a pharmaceutical composition for treating a disease alleviated by inhibiting the activity of p38α MAPK, comprising a therapeutically effective amount of a p38α MAPK inhibitor, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof, and a physiologically compatible carrier vehicle, for treating a disease alleviated by inhibiting the activity of p38α MAPK in a patient in need thereof, wherein the p38α MAPK inhibitor is a compound capable of binding to a pocket near the ED substrate docking site of p38α MAPK. In one embodiment, the binding pocket is defined by at least residues R49, H107, L108, and K165 of p38α MAPK. In one embodiment, the binding pocket is defined by residues R49, H107, L108, M109, G110, A157, V158, E163, L164, and K165 of p38α MAPK. In one embodiment, the p38α MAPK inhibitor is a compound of any of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly any of Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, or prodrug thereof. In some embodiments, the p38α MAPK inhibitor is a selective inhibitor of p38α MAPK.

[0141] In one embodiment, the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a p38α MAPK inhibitor, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof, and a physiologically compatible carrier medium for treating a disease alleviated by inhibiting the activity of p38α MAPK in a patient in need thereof, wherein the disease is cancer or an inflammatory disease. In one embodiment, the disease is rheumatoid arthritis, cardiovascular disease, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), asthma, acute respiratory distress syndrome (ARDS), or acute lung injury (ALI). In one embodiment, the disease is acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, cholangiocarcinoma, bladder cancer, brain cancer, breast cancer, bronchogenic carcinoma, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngeal carcinoma, cystadenocarcinoma, embryonal carcinoma, endothelioma, ependymoma, epithelial carcinoma, esophageal carcinoma, Ewing's tumor, fibrosarcoma, gastric cancer, glioblastoma multiforme, glioma, head and neck cancer. , hemangioblastoma, hepatocellular carcinoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendothelial sarcoma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteogenic sarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, Cancers include sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, gastric cancer, sweat gland carcinoma, synovium, testicular cancer, small cell lung cancer, pharyngeal cancer, uterine cancer, Wilms' tumor, blood cancer, acute erythroleukemia, acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monoblastic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute nonlymphocytic leukemia, acute promyelocytic leukemia, acute anaplastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin's lymphoma, polycythemia vera, and Waldenstrom's macroglobulinemia.

[0142] In some embodiments, the active pharmaceutical ingredient provided in the pharmaceutical composition of the present invention, e.g., any of the active pharmaceutical ingredients among the p38α MAPK inhibitors of the present invention, e.g., any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly any of Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044), may be present in, for example, 100%, 90%, 80%, 95%, 100% or more of the pharmaceutical composition. 0%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02% , 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or less than 0.0001% (w / w, w / v, or v / v).

[0143] In some embodiments, the active pharmaceutical ingredient provided in the pharmaceutical composition of the present invention, e.g., any of the p38α MAPK inhibitors of the present invention, e.g., any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-01 The concentration of each of the active pharmaceutical ingredients of any of SF-6-217, SF-6-223, SF-6-224, SF-6-219, SF-6-222, SF-7-009, and SF-7-044 is 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19.75%, 19.50%, 19.25%, 20% or 25% of the pharmaceutical composition. 19%, 18.75%, 18.50%, 18.25% 18%, 17.75%, 17.50%, 17.25% 17%, 16.75%, 16.50%, 16.25% 16%, 15.75%, 15.50%, 15.25% 15%, 14.75%, 14.50%, 14.25% 14%, 13.75%, 13.50%, 13.25% 13%, 12.75%, 12.50%, 12.25% 12%, 11.75%, 11.50%, 11.25% 11%, 10.75%, 10.50%, 10.25% 10%, 9.75%, 9.50%, 9.25% 9%, 8.75%, 8.50%, 8.25% 8%, 7.75%, 7.50%, 7.25% 7%, 6.75%, 6.50%, 6.25% 6%, 5.75%, 5.50%, 5.25% 5%,4.75%,4.50%,4.25%,4%,3.75%,3.50%,3.25%,3%,2.75%,2.50%,2.25%,2%,1.75%,1.50%,125%,1%,0.5%,0.4%,0.3%,0.2%,0.1%,0.09%,0.08%,0.07%,0.06%,0.05%,0.04%,0 .03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.More than 0.001% (w / w, w / v, or v / v).

[0144] In some embodiments, the active pharmaceutical ingredient provided in the pharmaceutical composition of the present invention, e.g., any of the active pharmaceutical ingredients of the p38α MAPK inhibitors of the present invention, e.g., any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044) The individual concentrations of any active pharmaceutical ingredient may be from about 0.0001% to about 50%, from about 0.001% to about 40%, from about 0.01% to about 30%, from about 0.02% to about 29%, from about 0.03% to about 28%, from about 0.04% to about 27%, from about 0.05% to about 26%, from about 0.06% to about 25%, from about 0.07% to about 24%, from about 0.08% to about 23% of the pharmaceutical composition. , about 0.09% to about 22%, about 0.1% to about 21%, about 0.2% to about 20%, about 0.3% to about 19%, about 0.4% to about 18%, about 0.5% to about 17%, about 0.6% to about 16%, about 0.7% to about 15%, about 0.8% to about 14%, about 0.9% to about 12%, or about 1% to about 10% (w / w, w / v, or v / v).

[0145] In some embodiments, the active pharmaceutical ingredient provided in the pharmaceutical composition of the present invention, e.g., any of the active pharmaceutical ingredients of the p38α MAPK inhibitors of the present invention, e.g., any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 106 The concentration of each of the active pharmaceutical ingredients of any of SF-1 (SF-6-219), SF-1085 (SF-6-222), SF-7-009, and SF-1087 (SF-7-044) is within the range of about 0.001% to about 10%, about 0.01% to about 5%, about 0.02% to about 4.5%, about 0.03% to about 4%, about 0.04% to about 3.5%, about 0.05% to about 3%, about 0.06% to about 2.5%, about 0.07% to about 2%, about 0.08% to about 1.5%, about 0.09% to about 1%, or about 0.1% to about 0.9% (w / w, w / v, or v / v) of the pharmaceutical composition.

[0146] In some embodiments, the active pharmaceutical ingredient provided in the pharmaceutical composition of the present invention is any of the aforementioned p38α MAPK inhibitors of the present invention, such as any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 ( The amount of each of the active pharmaceutical ingredients in any of SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044) is 10g, 9.5g, 9.0g, 8.5g, 8.0g, 7.5g, 7.0g, 6.5g, and 6. 0g, 5.5g, 5.0g, 4.5g, 4.0g, 3.5g, 3.0g, 2.5g, 2.0g, 1.5g, 1.0g, 0.95g, 0.9g, 0.85g, 0.8g, 0.75g, 0.7g, 0 .65g, 0.6g, 0.55g, 0.5g, 0.45g, 0.4g, 0.35g, 0.3g, 0.25g, 0.2g, 0.15g, 0.1g, 0.09g, 0.08g, 0.07g, 0.06 g, 0.05g, 0.04g, 0.03g, 0.02g, 0.01g, 0.009g, 0.008g, 0.007g, 0.006g, 0.005g, 0.004g, 0.003g, 0.002g, 0.001g, 0.0009g, 0.0008g, 0.0007g, 0.0006g, 0.0005g, 0.0004g, 0.0003g, 0.0002g, or 0.0001g.

[0147] In some embodiments, the active pharmaceutical ingredient provided in the pharmaceutical composition of the invention is any of the p38α MAPK inhibitors of the invention, such as any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-218), 1044 (SF-6-221), 1045 (SF-6-222), 1046 (SF-6-218), 1047 (SF-6-218), 1048 (SF-6-221), 1049 (SF-6-221), 1050 (SF-6-221), 1051 (SF-6-221), 1052 (SF-6-221), 1053 (SF-6-221), 1054 (SF-6-221), 1055 (SF-6-221), 1056 (SF-6-221), 1057 (SF-6-221), 1058 (SF-6-221), 1059 (SF-6-221), 1060 (SF-6-221), 1061 (SF-6-221), 1062 (SF-6-221), 1063 (SF-6-221), 1064 (SF-6-221), 1065 (SF-6-221), 1066 (SF-6-221), 1067 (SF-6-221), 1068 (SF-6-221 The amount of each of the active pharmaceutical ingredients of any of SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044) is 0.0001g, 0.0002g, 0.0003g, 0.0004g, 0.0005g, 0.0006g, 0.0007g, 0.0008g, 0.0009g, 0.001g, 0.0015g, 0.002g, 0.0025 ... g,0.003g,0.0035g,0.004g,0.0045g,0.005g,0.0055g,0.006g,0.0065g,0.007g,0.0075g,0.008g,0.0085g,0.009g,0. 0095g, 0.01g, 0.015g, 0.02g, 0.025g, 0.03g, 0.035g, 0.04g, 0.045g, 0.05g, 0.055g, 0.06g, 0.065g, 0.07g, 0.075g, 0.08 g, 0.085g, 0.09g, 0.095g, 0.1g, 0.15g, 0.2g, 0.25g, 0.3g, 0.35g, 0.4g, 0.45g, 0.5g, 0.55g, 0.6g, 0.65g, 0.7g, 0.75g, 0.8g, 0.85g, 0.9g, 0.95g, 1g, 1.5g, 2g, 2.5g, 3g, 3.5g, 4g, 4.5g, 5g, 5.5g, 6g, 6.5g, 7g, 7.5g, 8g, 8.5g, 9g, 9.5g, or greater than 10g.

[0148] Each of the active pharmaceutical ingredients according to the present invention is effective over a wide dosage range. For example, in the treatment of adult humans, dosages ranging from 0.01 to 1000 mg, 0.5 to 100 mg, 1 to 50 mg, and 5 to 40 mg per day are examples of dosages that may be used. The exact dosage will depend on the route of administration, the form in which the compound is administered, the gender and age of the subject being treated, the weight of the subject being treated, and the preference and experience of the attending physician. Clinically established dosages of the p38α MAPK inhibitors of the present invention may also be used, if desired.

[0149] In one embodiment, the molar ratio of the two active pharmaceutical ingredients in the pharmaceutical composition ranges from 10:1 to 1:10, preferably from 2.5:1 to 1:2.5, more preferably about 1:1. In one embodiment, the weight ratio of the molar ratio of the two active pharmaceutical ingredients in the pharmaceutical composition is selected from the group consisting of 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, and 1:20. In one embodiment, the weight ratio of the molar ratio of the two active pharmaceutical ingredients in the pharmaceutical composition is selected from the group consisting of 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, and 1:20.

[0150] In one embodiment, a pharmaceutical composition described herein, e.g., any of the p38α MAPK inhibitors of the present invention, e.g., any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly any of Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044), is used to treat an inflammatory disease. In one embodiment, the pharmaceutical compositions described herein (e.g., any of the p38α MAPK inhibitors of the present invention) are used to treat rheumatoid arthritis, cardiovascular disease, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), asthma, acute respiratory distress syndrome (ARDS), or acute lung injury (ALI).

[0151] In one embodiment, a pharmaceutical composition described herein, e.g., any of the p38α MAPK inhibitors of the present invention, e.g., any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly any of Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044), is used to treat a hyperproliferative disorder associated with overexpression or upregulation and / or downregulation of p38α MAPK protein.In some embodiments, the pharmaceutical compositions described herein are useful for treating cancers associated with overexpression or upregulation and / or downregulation of p38α MAPK protein (e.g., pancreatic cancer, breast cancer, prostate cancer, lymphoma, skin cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain cancer, head-neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck carcinoma, breast cancer, ovarian carcinoma, lung cancer, small cell lung cancer, Wilms' tumor, cervical cancer, testicular cancer, bladder carcinoma, pancreatic carcinoma, gastric cancer, colon carcinoma, prostate cancer, etc.). carcinoma), genitourinary cancer, thyroid cancer, esophageal cancer, myeloma, multiple myeloma, adrenal cancer, renal cell carcinoma, endometrial cancer, adrenocortical carcinoma, malignant pancreatic insulinoma, malignant carcinoid cancer, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic granulocytic leukemia, acute granulocytic leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, polycythemia vera, essential thrombocytosis, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, osteosarcoma, primary macroglobulinemia, or retinoblastoma).

[0152] Listed below are non-limiting pharmaceutical compositions and methods for their preparation.

[0153] Pharmaceutical Composition for Oral Administration In one embodiment, the present invention provides a pharmaceutical composition for oral administration, comprising an active pharmaceutical ingredient or a combination of active pharmaceutical ingredients, such as a p38α MAPK inhibitor as described herein, and a pharmaceutical excipient suitable for oral administration.

[0154] In some embodiments, the present invention provides solid pharmaceutical compositions for oral administration, comprising (i) an effective amount of an active pharmaceutical ingredient or combination of active pharmaceutical ingredients, and (ii) a pharmaceutical excipient suitable for oral administration. In selected embodiments, the composition further comprises (iii) an effective amount of a third active pharmaceutical ingredient, and optionally, (iv) an effective amount of a fourth active pharmaceutical ingredient.

[0155] In some embodiments, the pharmaceutical composition may be a liquid pharmaceutical composition suitable for oral ingestion. Pharmaceutical compositions of the present invention suitable for oral administration can be presented in separate dosage forms, such as capsules, sachets, or tablets, or as liquids or aerosol sprays, each containing a predetermined amount of the active ingredient as a powder, or in granules, solutions, or suspensions in aqueous or non-aqueous liquids, oil-in-water emulsions, water-in-oil liquid emulsions, powders for reconstitution, powders for oral ingestion, bottles (containing powders or liquids in bottles), orally dissolvable films, lozenges, pastes, tubes, gums, and packs. Such dosage forms can be prepared by any of the pharmaceutical methods, but all methods include the step of bringing the active ingredient(s) into association with a carrier, which constitutes one or more necessary ingredients. Generally, the compositions are prepared by uniformly and intimately admixing the active ingredient(s) with a liquid carrier or finely divided solid carrier, or both, and then, if necessary, shaping the product into the desired presentation. For example, tablets can be prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in free form (such as a powder or granules), optionally mixed with excipients (such as, but not limited to, binders, lubricants, inert diluents, and / or surface-active or dispersing agents). Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

[0156] Because water can accelerate the degradation of some compounds, the present invention further encompasses anhydrous pharmaceutical compositions and dosage forms. Pharmaceutical techniques may add water (e.g., 5%) as a means of simulating long-term storage to determine characteristics such as shelf life or the stability of a formulation over time. The anhydrous pharmaceutical compositions and dosage forms of the present invention can be prepared using anhydrous or low-moisture ingredients and low-moisture or low-humidity conditions. Pharmaceutical compositions and dosage forms of the present invention containing lactose can be made anhydrous if they are expected to be in substantial contact with moisture and / or humidity during manufacture, packaging, and / or storage. Anhydrous pharmaceutical compositions can be prepared and stored to maintain their anhydrous nature. Therefore, anhydrous compositions can be packaged using known materials to prevent exposure to water, and they can be included in suitable formulation kits. Examples of suitable packaging include, but are not limited to, sealed foils, sealed plastics, unit-dose containers, blister packs, and strip packs.

[0157] Each active pharmaceutical ingredient can be intimately mixed and combined with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. Carriers can take a variety of forms depending on the desired preparation form for administration. When preparing compositions for oral dosage forms, any common pharmaceutical medium can be used as a carrier. For example, for oral liquid preparations (such as suspensions, solutions, and elixirs) or aerosols, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, etc. can be used as carriers; for oral solid preparations, starch, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, etc. can be used as carriers; in some embodiments, lactose is not required. For example, suitable carriers include powders, capsules, and tablets, including solid oral preparations. If necessary, tablets can be coated by standard aqueous or non-aqueous techniques.

[0158] Binders suitable for use in pharmaceutical compositions and dosage forms include, but are not limited to, corn starch, potato starch, or other starches, gelatin, natural and synthetic gums (e.g., acacia), sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, calcium carboxymethylcellulose, sodium carboxymethylcellulose), polyvinylpyrrolidone, methylcellulose, pregelatinized starch, hydroxypropyl methylcellulose, microcrystalline cellulose, and mixtures thereof.

[0159] Examples of suitable fillers for use in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof.

[0160] Disintegrants can be used in the compositions of the present invention to provide tablets that disintegrate when exposed to an aqueous environment. Too much disintegrant can result in tablets that disintegrate in the bottle. Too little can be insufficient to cause disintegration, thus altering the rate and extent of release of the active ingredient from the dosage form. Therefore, a dosage form of the compound disclosed herein can be formed using a sufficient amount of disintegrant that is neither too little nor too much so as not to adversely alter the release of the active ingredient(s). The amount of disintegrant used can vary based on the type of formulation and mode of administration and is readily understood by those skilled in the art. About 0.5 to about 15% by weight of disintegrant, or about 1 to about 5% by weight of disintegrant, can be used in the pharmaceutical composition. Disintegrants that can be used to form pharmaceutical compositions and dosage forms of the invention include, but are not limited to, agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pregelatinized starch, other starches, clays, other algins, other celluloses, gums, or mixtures thereof.

[0161] Lubricants that can be used to form the pharmaceutical compositions and dosage forms of the present invention include, but are not limited to, calcium stearate, magnesium stearate, sodium stearyl fumarate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, or mixtures thereof. Additional lubricants include, for example, syloid silica gel, coagulated aerosol of synthetic silica, silicified microcrystalline cellulose, or mixtures thereof. Lubricants can optionally be added in an amount of less than about 0.5% by weight or less than about 1% by weight of the pharmaceutical composition.

[0162] When aqueous suspensions and / or elixirs are desired for oral administration, the active pharmaceutical ingredient(s) may be combined with various sweetening or flavoring agents, coloring agents or pigments, and, if necessary, emulsifying and / or suspending agents, diluents such as water, ethanol, propylene glycol, glycerin, and various combinations thereof.

[0163] Tablets may be uncoated or may be coated by known techniques, which delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained effect over a longer period of time.For example, time-delay materials such as glyceryl monostearate or glyceryl distearate can be used.Also, the formulation for oral use can be presented as a hard gelatin capsule in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate or kaolin, or as a soft gelatin capsule in which the active ingredient is mixed with an aqueous medium or an oil medium (for example, peanut oil, liquid paraffin or olive oil).

[0164] Surfactants that can be used to form the pharmaceutical compositions and dosage forms of the present invention include, but are not limited to, hydrophilic surfactants, lipophilic surfactants, and mixtures thereof, i.e., a mixture of hydrophilic surfactants can be used, a mixture of lipophilic surfactants can be used, or a mixture of at least one hydrophilic surfactant and at least one lipophilic surfactant can be used.

[0165] Suitable hydrophilic surfactants generally have an HLB value of at least 10, while suitable lipophilic surfactants generally have an HLB value of about 10 or less. The empirical parameter used to characterize the relative hydrophilicity and relative hydrophobicity of nonionic amphiphilic compounds is the hydrophilic-lipophilic balance ("HLB" value). Surfactants with lower HLB values ​​are more lipophilic or hydrophobic and have higher solubility in oils, while surfactants with higher HLB values ​​are more hydrophilic and have higher solubility in aqueous solutions. Hydrophilic surfactants are generally considered to be compounds with an HLB value greater than about 10, as well as anionic, cationic, or zwitterionic compounds to which the HLB scale does not generally apply. Similarly, lipophilic (i.e., hydrophobic) surfactants are compounds with an HLB value of about 10 or less. However, the HLB value of a surfactant is generally only a rough guide used to enable the formulation of emulsions for industrial, pharmaceutical, and cosmetic applications.

[0166] Hydrophilic surfactants can be either ionic or nonionic.Suitable ionic surfactants include but are not limited to alkylammonium salts, fusidate salts, fatty acid derivatives of amino acids, oligopeptides and polypeptides, glyceride derivatives of amino acids, oligopeptides and polypeptides, lecithin and hydrogenated lecithin, lysolecithin and hydrogenated lysolecithin, phospholipids and their derivatives, lysophospholipids and their derivatives, carnitine fatty acid ester salts, alkyl sulfate salts, fatty acid salts, docusate sodium, acyl lactylates, mono- and diacetylated tartaric acid esters of mono- and diglycerides, succinylated mono- and diglycerides, citrate esters of mono- and diglycerides, and mixtures thereof.

[0167] Among the aforementioned groups, examples of ionic surfactants include lecithin, lysolecithin, phospholipids, lysophospholipids, and derivatives thereof, carnitine fatty acid ester salts, alkyl sulfates, fatty acid salts, sodium docusate, acyl acrylates, mono- and diacetylated tartaric acid esters of mono- and diglycerides, succinylated mono- and diglycerides, citrate esters of mono- and diglycerides, and mixtures thereof.

[0168] Ionic surfactants include lecithin, lysolecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylserine, PEG-phosphatidylethanolamine, PVP-phosphatidylethanolamine, and lactylic acid esters of fatty acids. The ionized forms may be esters of glycerin, stearoyl-2-lactylate, stearoyl lactylate, succinylated monoglycerides, mono / diacetylated tartaric acid esters of mono / diglycerides, citrate esters of mono / diglycerides, cholyl sarcosine, caproate, caprylate, caprate, laurate, myristate, palmitate, oleate, ricinoleate, linoleate, linolenate, stearate, lauryl sulfate, teracecyl sulfate, docusate, lauroylcarnitine, palmitoylcarnitine, myristoylcarnitine, and mixtures thereof.

[0169] Examples of hydrophilic nonionic surfactants include alkyl glucosides, alkyl maltosides, alkyl thioglucosides, lauryl macrogol glycerides, polyoxyalkylene alkyl ethers (e.g., polyethylene glycol alkyl ethers), polyoxyalkylene alkylphenols (e.g., polyethylene glycol alkylphenols), polyoxyalkylene alkylphenol fatty acid esters (e.g., polyethylene glycol fatty acid monoesters and polyethylene glycol fatty acid diesters), polyethylene glycol glycerol fatty acid esters, polyglycerol fatty acid esters, polyoxyalkylene sorbitan fatty acid esters (e.g., polyethylene glycol Examples of suitable polyols include, but are not limited to, glycerol sorbitan fatty acid esters, hydrophilic transesterification products of polyols (including at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids, and sterols), polyoxyethylene sterols (including their derivatives and analogs), polyoxyethylated vitamins and their derivatives, polyoxyethylene-polyoxypropylene block copolymers and mixtures thereof, polyethylene glycol sorbitan fatty acid esters, and hydrophilic transesterification products of polyols (including at least one member of the group consisting of triglycerides, vegetable oils, and hydrogenated vegetable oils). The polyol may be glycerol, ethylene glycol, polyethylene glycol, sorbitol, propylene glycol, pentaerythritol, or a sugar.

[0170] Other hydrophilic nonionic surfactants include PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32 dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate, PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32 distearate, PEG-40 stearate, and PEG-100 stearate. Salt, PEG-20 Dilaurate, PEG-25 Glyceryl Trioleate, PEG-32 Dioleate, PEG-20 Glyceryl Laurate, PEG-30 Glyceryl Laurate, PEG-20 Glyceryl Stearate, PEG-20 Glyceryl Oleate, PEG-30 Glyceryl Oleate, PEG-30 Glyceryl Laurate, PEG-40 Glyceryl Laurate, PEG-40 Palm Kernel Oil, PEG-50 Hydrogenated Castor Oil, PEG-40 Castor Oil, PEG-35 Castor Oil, PEG-60 Castor Oil, PEG-40 Hydrogenated Castor Oil, PEG-60 Hydrogenated Castor Oil, PEG-60 Corn Oil, PEG-6 Capric / Caprylic Glycerides, PEG-8 Capric / Caprylic Glycerides, Polyglyceryl-10 Laurate, PEG-30 Cholesterol, PEG-25 Phytosterols, PEG-30 Soy Sterols, PEG-20 Trioleate, PEG-40 Sorbitan Oleate, PEG-80 Sorbitan Laurate, Polysorbate 20, Polysorbate 80, POE-9 Lauryl Ether, POE- Examples of suitable glycerin-based copolymers include, but are not limited to, PEG-23 lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearyl ether, tocopheryl PEG-100 succinate, PEG-24 cholesterol, polyglyceryl-10 oleate, Tween 40, Tween 60, sucrose monostearate, sucrose monolaurate, sucrose monopalmitate, PEG10-100 nonylphenol series, PEG15-100 octylphenol series, and poloxamer.

[0171] Suitable lipophilic surfactants include, by way of example only, fatty alcohols, glycerol fatty acid esters, acetylated glycerol fatty acid esters, lower alcohol fatty acid esters, propylene glycol fatty acid esters, sorbitan fatty acid esters, polyethylene glycol sorbitan fatty acid esters, sterols and sterol derivatives, polyoxyethylated sterols and sterol derivatives, polyethylene glycol alkyl ethers, sugar esters, sugar ethers, lactic acid derivatives of mono- and diglycerides, hydrophobic transesterification products of polyols (including at least one member of the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols), oil-soluble vitamins / vitamin derivatives, and mixtures thereof.In this group, preferred lipophilic surfactants include glycerol fatty acid esters, propylene glycol fatty acid esters, and mixtures thereof, or hydrophobic transesterification products of polyols, including at least one member of the group consisting of vegetable oils, hydrogenated vegetable oils, and triglycerides.

[0172] In one embodiment, the composition may contain a solubilizing agent to ensure good solubilization and / or dissolution of the compound of the present invention and minimize precipitation of the compound of the present invention. This may be particularly important for compositions for parenteral use (e.g., injectable compositions). A solubilizing agent may also be added to increase the solubility of hydrophilic drugs and / or other components (such as surfactants) or to maintain the composition as a stable or homogeneous solution or dispersion.

[0173] Examples of suitable solubilizing agents include, but are not limited to, alcohols and polyols (e.g., ethanol, isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene glycol, butanediol and its isomers, glycerol, pentaerythritol, sorbitol, mannitol, transcutol, dimethyl isosorbide, polyethylene glycol, polypropylene glycol, polyvinyl alcohol, hydroxypropyl methylcellulose and other cellulose derivatives, cyclodextrin and cyclodextrin derivatives), ethers of polyethylene glycol having an average molecular weight of about 200 to about 6000 (e.g., tetrahydrofurfuryl alcohol PEG ether (glycofurol) or methoxy PEG), amides and other nitrogen-containing compounds (e.g., 2-pyrrolidone, 2-piperidine diphosphate), and the like. Examples of suitable solubilizing agents include propylene glycol monoacetate, propylene glycol diacetate, propylene glycol hydroxypropyl ether, propylene glycol methyl ...

[0174] Mixtures of solubilizers can also be used. Examples include, but are not limited to, triacetin, triethyl citrate, ethyl oleate, ethyl caprylate, dimethylacetamide, N-methylpyrrolidone, N-hydroxyethylpyrrolidone, polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxypropyl cyclodextrin, ethanol, polyethylene glycol 200-100, glycofurol, transcutol, propylene glycol, and dimethyl isosorbide. Particularly preferred solubilizers include sorbitol, glycerol, triacetin, ethyl alcohol, PEG-400, glycofurol, and propylene glycol.

[0175] The amount of solubilizer that can be included is not particularly limited. The amount of a given solubilizer may be limited to a biotolerable amount, which can be easily determined by one skilled in the art. In some situations, for example, to maximize the drug concentration, it may be advantageous to include an amount of solubilizer that far exceeds the biotolerable amount and remove the excess solubilizer using conventional techniques such as distillation or evaporation before providing the composition to a patient. Thus, when present, the solubilizer may be present in a weight ratio of 10%, 25%, 50%, 100%, or up to about 200% by weight based on the combined weight of the drug and other excipients. If desired, very small amounts of solubilizer, such as 5%, 2%, 1%, or even less, may be used. Typically, the solubilizer may be present in an amount of about 1% to about 100% by weight, more typically about 5% to about 25% by weight.

[0176] The composition can further comprise one or more pharmaceutically acceptable additives and excipients, including, but not limited to, release agents, antifoaming agents, buffers, polymers, antioxidants, preservatives, chelating agents, viscosity modifiers, tonicity agents, flavors, colorants, odorants, opacifying agents, suspending agents, binders, fillers, plasticizers, lubricants, and mixtures thereof.

[0177] In addition, acids or bases can be incorporated into the composition to facilitate processing, enhance stability, or for other reasons. Examples of pharmaceutically acceptable bases include amino acids, amino acid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide, sodium bicarbonate, aluminum hydroxide, calcium carbonate, magnesium hydroxide, magnesium aluminum silicate, synthetic aluminum silicate, synthetic hydrocalcite, magnesium aluminum hydroxide, diisopropylethylamine, ethanolamine, ethylenediamine, triethanolamine, triethylamine, triisopropanolamine, trimethylamine, tris(hydroxymethyl)aminomethane (TRIS), etc. Also suitable are bases that are salts of pharmaceutically acceptable acids (e.g., acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acids, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid, hydroquinosulfonic acid, isoascorbic acid, lactic acid, maleic acid, oxalic acid, para-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, uric acid, and the like). Salts of polybasic acids, such as sodium phosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphate, can also be used. When the base is a salt, the cation can be any convenient, pharmaceutically acceptable cation, such as ammonium, alkali metals, and alkaline earth metals. Examples can include, but are not limited to, sodium, potassium, lithium, magnesium, calcium, and ammonium.

[0178] Suitable acids are pharmaceutically acceptable organic or inorganic acids. Examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, boric acid, phosphoric acid, etc. Examples of suitable organic acids include acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acid, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid, hydroquinosulfonic acid, isoascorbic acid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid, para-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, and uric acid.

[0179] Injectable pharmaceutical composition In some embodiments, a pharmaceutical composition is provided for injection and comprises an active pharmaceutical ingredient (such as a p38α MAPK inhibitor, e.g., any of the compounds of Formula A, Formula I, Formula II, any of Formulas 1001-1180, particularly any of Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044)), or a combination of active pharmaceutical ingredients, and a pharmaceutical excipient suitable for injection.

[0180] Forms into which the compositions of the present invention may be incorporated for administration by injection include aqueous or oily suspensions, or emulsions (with sesame, corn, cottonseed, or peanut oil, as well as elixirs, mannitol, dextrose, or sterile aqueous solutions, and similar pharmaceutical vehicles).

[0181] Conventionally, aqueous solutions in physiological saline are also used for injection. Ethanol, glycerol, propylene glycol and liquid polyethylene glycol (and their suitable mixtures), cyclodextrin derivatives, and vegetable oils can also be used. Appropriate fluidity can be maintained, for example, by using coating agents (such as lecithin, to maintain the required particle size in the case of dispersions) and surfactants. Prevention of microbial action can be achieved by various antibacterial and antifungal agents (for example, parabens, chlorobutanol, phenol, sorbic acid, and thimerosal).

[0182] Sterile injectable solution can be prepared by incorporating active pharmaceutical ingredients or a combination of active pharmaceutical ingredients into a suitable solvent containing various other ingredients listed above in the required amount, and then sterilizing by filtration.Generally, dispersion is prepared by incorporating various sterilized active ingredients into a sterile vehicle that contains basic dispersion medium and other necessary ingredients listed above.For the sterile powder used to prepare sterile injectable solution, the specific desired preparation method is vacuum drying and freeze-drying technology, and from the solution that has been previously sterilized and filtered, powder of active ingredients and any other desired ingredients can be obtained.

[0183] Pharmaceutical Compositions for Topical Delivery In some embodiments, a pharmaceutical composition is provided for transdermal delivery and includes an active pharmaceutical ingredient (such as a p38α MAPK inhibitor described herein, e.g., any of the compounds of Formula A, Formula I, Formula II, any of Formulas 1001-1180, particularly any of Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044)), or a combination of active pharmaceutical ingredients, and a pharmaceutical excipient suitable for transdermal delivery.

[0184] The composition of the present invention can be formulated into solid, semi-solid or liquid form preparations suitable for local or topical administration, such as gel, water-soluble jelly, cream, lotion, suspension, foam, powder, slurry, ointment, solution, oil, paste, suppository, spray, emulsion, saline solution, dimethyl sulfoxide (DMSO)-based solution, etc. Generally, carriers with higher density can provide a certain area with prolonged exposure to active ingredients.In contrast, solution formulations can provide a more immediate exposure of active ingredients to selected areas.

[0185] Pharmaceutical compositions can also contain suitable solid or gel phase carriers or excipients, which are compounds that increase the permeability of therapeutic molecules across the permeability barrier of the stratum corneum of the skin or assist in the delivery of therapeutic molecules.There are many such penetration-enhancing molecules, and they are known to those skilled in the art of topical formulations.Examples of such carriers and excipients include, but are not limited to, moisturizers (e.g., urea), glycols (e.g., propylene glycol), alcohols (e.g., ethanol), fatty acids (e.g., oleic acid), surfactants (e.g., isopropyl myristate and sodium lauryl sulfate), pyrrolidone, glycerol monolaurate, sulfoxides, terpenes (e.g., menthol), amines, amides, alkanes, alkanols, water, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers (such as polyethylene glycol).

[0186] Another exemplary formulation for use in the methods of the present invention employs transdermal delivery devices ("patches"). Such transdermal patches can be used to provide continuous or discontinuous infusion of an active pharmaceutical ingredient or a combination of active pharmaceutical ingredients, with or without another active pharmaceutical ingredient, in controlled amounts.

[0187] The construction and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. See, for example, U.S. Patent Nos. 5,023,252, 4,992,445, and 5,001,139 (the entireties of which are incorporated herein by reference). Such patches can be constructed for continuous, pulsatile, or on-demand delivery of pharmaceutical agents.

[0188] Pharmaceutical Composition for Inhalation Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders. The liquid or solid compositions may include a suitable pharmaceutically acceptable excipient as described above and a p38α MAPK inhibitor described herein, such as any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly any of the inhibitors of Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044). Preferably, the composition is administered by oral or nasal respiratory route for local or systemic effect.Preferably, the composition in a pharmaceutically acceptable solvent can be atomized by using inert gas.Atomized solution can be inhaled directly from the atomizing device, or the atomizing device can be attached to a face mask tent or an intermittent positive pressure breathing machine.Solution, suspension, or powder composition can be administered from the device that delivers the formulation in an appropriate manner, preferably orally or nasally.Dry powder inhalers can also be used to provide the inhalation delivery of the composition.

[0189] Other pharmaceutical compositions Pharmaceutical compositions of the p38α MAPK inhibitors described herein, for example, any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-224), 1044 (SF-6-225), 1045 (SF-6-226), 1046 (SF-6-227), 1047 (SF-6-228), 1048 (SF-6-229), 1049 (SF-6-230), 1050 (SF-6-231), 1051 (SF-6-232), 1052 (SF-6-233), 1053 (SF-6-234), 1054 (SF-6-235), 1055 (SF-6-236), 1056 (SF-6-237), 1057 (SF-6-238), 1058 (SF-6-239), 1059 (SF-6-239), 1060 (SF-6-239), 1061 (SF-6-239), 1062 (SF-6-239), 1063 (SF-6-239), 1064 (SF-6-239), 1065 (SF-6-239), 1066 (SF-6-239), 1067 (SF-6-239), 1068 (SF-6-239), 1069 (SF-6-239), Pharmaceutical compositions of any of the inhibitors of SF-23), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044) can also be prepared from the compositions described herein and one or more pharmaceutically acceptable excipients suitable for sublingual, buccal, rectal, intraosseous, intraocular, intranasal, epidural, or intraspinal administration. The preparation of such pharmaceutical compositions is well known in the art. See, for example, Anderson, Philip O.; Knoben, James E.; Troutman, William G, eds., Handbook of Clinical Drug Data, Tenth Edition, McGraw-Hill, 2002, and Pratt and Taylor, eds., Principles of Drug Action, Third Edition, Churchill Livingston, NY, 1990, each of which is incorporated herein by reference in its entirety.

[0190] The administration of the active pharmaceutical ingredient or the combination of active pharmaceutical ingredients, or its pharmaceutical composition, can be carried out by any method that allows the compound to be delivered to the site of action.These methods include oral route, intraduodenal route, parenteral injection (including intravenous, intraarterial, subcutaneous, intramuscular, intravascular, intraperitoneal, or infusion), topical (e.g., transdermal application), rectal administration, local delivery by catheter or stent, or inhalation.The active pharmaceutical ingredient or the combination of active pharmaceutical ingredients can also be administered intraadiposely or intrathecally.

[0191] Exemplary parenteral administration forms include solutions or suspensions of the active compounds in sterile aqueous solutions, for example, aqueous propylene glycol or dextrose. Such dosage forms may be suitably buffered, if necessary.

[0192] kit The present invention also provides kits. The kits include an active pharmaceutical ingredient or combination of active pharmaceutical ingredients (either alone or in suitable packaging) and written materials that may include instructions for use, clinical trial discussions, and a list of side effects. Such kits may also include information such as scientific references, package inserts, clinical trial results, and / or summaries thereof, demonstrating or establishing the activity and / or benefits of the composition, and / or describing dosage, administration, side effects, drug interactions, or other information useful to healthcare providers. Such information may be based on the results of various studies (e.g., studies using laboratory animals, including in vivo models, and studies based on human clinical trials). The kits may further include another active pharmaceutical ingredient. In selected embodiments, the active pharmaceutical ingredient or combination of active pharmaceutical ingredients is provided as a separate composition in a separate container within the kit. In selected embodiments, the active pharmaceutical ingredient or combination of active pharmaceutical ingredients is provided as a single composition within the kit container. Suitable packaging and additional items for use (e.g., measuring cups for liquid preparations, foil packaging to minimize exposure to air, etc.) are known in the art and may be included in the kit. The kits described herein may be provided, marketed, and / or promoted to healthcare providers (including physicians, nurses, pharmacists, prescribers, etc.) The kits may also, in selected embodiments, be sold directly to consumers.

[0193] In some embodiments, the present invention provides kits, comprising a therapeutically effective amount of an active pharmaceutical ingredient (e.g., a p38α MAPK inhibitor, e.g., any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-7-014), 1038 (SF-7-016), 1039 (SF-7-019), 1040 (SF-7-019), 1041 (SF-7-020), 1042 (SF-7-023), 1043 (SF-7-024), 1044 (SF-7-025), 1045 (SF-7-026), 1046 (SF-7-027), 1047 (SF-7-028), 1048 (SF-7-029), 1049 (SF-7-030), 1050 (SF-7-031), 1051 (SF-7-032), 1052 (SF-7-033), 1053 (SF-7-034), 1054 (SF-7-035), 1055 (SF-7-036), 1056 (SF-7-037), 1057 (SF-7-038), 1058 (SF-7-039), 1060 (SF-7-039), 1061 (SF-7-039), 1062 (SF-7-039), 1063 (SF-7-039), 1064 The kit includes a composition comprising a combination of active pharmaceutical ingredients, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof, of any of the compounds selected from the group consisting of SF-6-217, 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044). These compositions are typically pharmaceutical compositions. The kit is for co-administration of the active pharmaceutical ingredients or combination of active pharmaceutical ingredients, either simultaneously or separately.

[0194] In some embodiments, the present invention provides kits, the kits including: (1) a therapeutically effective amount of an active pharmaceutical ingredient (e.g., a p38α MAPK inhibitor, e.g., any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (1) a composition comprising one or more of the active pharmaceutical ingredients selected from the group consisting of SF-6-223, 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044), or a combination of active pharmaceutical ingredients, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof; and (2) a diagnostic test for determining whether a patient's cancer is a specific subtype of cancer. Any of the foregoing diagnostic methods can be utilized in a kit.

[0195] The kit described above is preferably used for the treatment of the diseases and conditions described herein. In some embodiments, the kit is used for the treatment of inflammatory diseases. In some embodiments, the kit is used for the treatment of rheumatoid arthritis, cardiovascular disease, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), asthma, acute respiratory distress syndrome (ARDS), or acute lung injury (ALI). In certain embodiments, the kit is used for the treatment of hyperproliferative disorders (e.g., cancer).

[0196] In certain embodiments, the kits described herein are used to treat cancer. In some embodiments, the kits described herein are used to treat pancreatic cancer, breast cancer, prostate cancer, lymphoma, skin cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain cancer, head-neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck cancer, breast cancer, ovarian carcinoma, lung cancer, small cell lung cancer, Wilms' tumor, cervical cancer, testicular cancer, bladder carcinoma, pancreatic carcinoma, gastric cancer, colon carcinoma, prostate cancer. carcinoma), genitourinary cancer, thyroid cancer, esophageal cancer, myeloma, multiple myeloma, adrenal cancer, renal cell carcinoma, endometrial cancer, adrenocortical carcinoma, malignant pancreatic insulinoma, malignant carcinoid cancer, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic granulocytic leukemia, acute granulocytic leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, polycythemia vera, essential thrombocytosis, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, osteosarcoma, primary macroglobulinemia, and retinoblastoma. In certain embodiments, the kits described herein are used to treat malignant melanoma.

[0197] Dosage and Dosage Regimen The amount of pharmaceutical composition administered using the methods herein, e.g., a p38α MAPK inhibitor, e.g., any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF The dosage of any of SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044) will depend on the human or mammal being treated, the severity of the disorder or condition, the rate of administration, the kinetics of the active pharmaceutical ingredient, and the discretion of the prescribing physician. However, effective dosages range from about 0.001 to about 100 mg per kg of body weight per day (e.g., about 1 to about 35 mg / kg / day), in single or divided doses. For a 70 kg human, this would amount to about 0.05 to 7 g / day (e.g., about 0.05 to about 2.5 g / day). In some cases, dosage levels below the lower end of the aforementioned ranges may be more than sufficient, while in other cases even larger doses may be used without causing any adverse side effects (e.g., by dividing such larger doses into several smaller doses to be administered throughout the day). Dosages of pharmaceutical compositions and active pharmaceutical ingredients are given in mg / kg body weight, or mg / m 2 It may be provided in units of body surface area.

[0198] In some embodiments, the present invention includes a method of treating cancer in a human subject suffering from a cancer in which the cancer cells overexpress p38α MAPK, the method comprising administering a therapeutically effective dose of an active pharmaceutical ingredient, i.e., a p38α MAPK inhibitor, such as any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 103 The method includes administering to a human subject an inhibitor of any of SF-6-4 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044).

[0199] In some embodiments, the present invention includes a method of treating cancer in a human subject suffering from a cancer in which the cancer cells overexpress p38α MAPK, the method comprising administering to a subject a therapeutically effective dose of an active pharmaceutical ingredient, i.e., a p38α MAPK inhibitor, such as any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), The method includes administering to a human subject an inhibitor of any of 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044) to inhibit or reduce the activity of p38α MAPK protein.

[0200] In some embodiments, the pharmaceutical composition or active pharmaceutical ingredient is administered in a single dose. Such administration can be by injection (e.g., intravenous injection) to rapidly introduce the active pharmaceutical ingredient. However, other routes (including the preferred oral route) can be used if necessary. A single dose of the pharmaceutical composition can also be used to treat acute conditions.

[0201] In some embodiments, the pharmaceutical composition or active pharmaceutical ingredient is administered in multiple doses. In one embodiment, the pharmaceutical composition is administered in multiple doses. Dosing can be once, twice, three times, four times, five times, six times, or more than six times daily. Dosing can be monthly, once every two weeks, once weekly, or once every other day. In other embodiments, the pharmaceutical composition is administered from about once to about six times daily. In some embodiments, the pharmaceutical composition is administered once daily, while in other embodiments, the pharmaceutical composition is administered twice daily, and in other embodiments, the pharmaceutical composition is administered three times daily.

[0202] Administration of the active pharmaceutical ingredient can continue for as long as necessary. In selected embodiments, the pharmaceutical composition is administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, the pharmaceutical composition is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day(s). In some embodiments, the pharmaceutical composition is administered chronically, for example, for the treatment of chronic effects. In some embodiments, administration of the pharmaceutical composition continues for less than about 7 days. In yet other embodiments, administration continues for more than about 6, 10, 14, 28 days, 2 months, 6 months, or 1 year. In some cases, continuous dosing is achieved and maintained for as long as necessary.

[0203] In some embodiments, an effective dosage of an active pharmaceutical ingredient disclosed herein, e.g., any of the compounds of Formula A, Formula I, Formula II, or any of Formulas 1001-1180, particularly any of Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044), is about 1 mg to about 500 mg, about 10 mg to about 300 mg, or about 20 mg to about 250 mg. , about 25 mg to about 200 mg, about 10 mg to about 200 mg, about 20 mg to about 150 mg, about 30 mg to about 120 mg, about 10 mg to about 90 mg, about 20 mg to about 80 mg, Approximately 30mg to approximately 70mg, approximately 40mg to approximately 60mg, approximately 45mg to approximately 55mg, approximately 48mg to approximately 52mg, approximately 50mg to approximately 150mg, approximately 60mg to approximately 140mg, approximately 70 In some embodiments, the effective dose of an active pharmaceutical ingredient disclosed herein is less than about 25 mg, less than about 50 mg, less than about 75 mg, less than about 100 mg, less than about 125 mg, less than about 150 mg, less than about 175 mg, less than about 200 mg, less than about 225 mg, or less than about 250 mg. In some embodiments, the effective dosage of an active pharmaceutical ingredient disclosed herein is greater than about 25 mg, greater than about 50 mg, greater than about 75 mg, greater than about 100 mg, greater than about 125 mg, greater than about 150 mg, greater than about 175 mg, greater than about 200 mg, greater than about 225 mg, or greater than about 250 mg.

[0204] In some embodiments, the active pharmaceutical ingredient disclosed herein, e.g., any of the compounds of Formula A, Formula I, Formula II, Formulas 1001-1180, particularly Formulas 1001 (SF-6-221), 1032 (SF-7-008), 1034 (SF-7-010), 1035 (SF-7-011), 1036 (SF-7-012), 1037 (SF-6-217), , 1043 (SF-6-223), 1049 (SF-6-224), 1061 (SF-6-219), 1085 (SF-6-222), SF-7-009, and 1087 (SF-7-044) may be administered in an amount ranging from about 0.01 to about 200 mg / kg, or from about 0.1 to 100 mg / kg, or from about 1 to 50 mg / kg.

[0205] In some embodiments, the active pharmaceutical ingredient is administered at a dosage of 10 to 200 mg twice daily (including 50, 60, 70, 80, 90, 100, 150, or 200 mg twice daily). In some embodiments, the active pharmaceutical ingredient is administered at a dosage of 10 to 500 mg twice daily (including 1, 5, 10, 15, 25, 50, 75, 100, 150, 200, 300, 400, or 500 mg twice daily).

[0206] In some cases, dosage levels below the lower limit of the aforementioned range may be more than sufficient, while in other cases, even larger doses may be used without causing adverse side effects (e.g., by dividing such larger doses into several smaller doses to be administered throughout the day). Of course, as those skilled in the art will understand, the actual dose administered will depend on the condition being treated, the age, health, and weight of the recipient, the type of concurrent treatment, if any, and the frequency of treatment. Furthermore, the effective dose can be determined by those skilled in the art based on routine empirical activity tests to measure the biological activity of the compound(s) in bioassays and thus establish the appropriate dose to be administered.

[0207] An effective amount of the combination of active pharmaceutical ingredients can be administered in either single or multiple doses by any of the accepted modes of administration for drugs having similar utilities, including rectal, buccal, intranasal and transdermal routes, by intraarterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, topically, or as inhalation.

[0208] In some embodiments, the compositions described herein further include controlled-release, sustained-release, or long-term release therapeutic dosage forms for administering the compounds described herein, which involve incorporating the compound into a suitable delivery system in the formulation of a particular composition. The dosage form controls the release of the compound(s) so that effective concentrations of the compound(s) in the bloodstream can remain relatively constant over time to improve therapeutic outcomes and / or minimize side effects. Furthermore, the controlled-release system will result in minimal peak-to-trough fluctuations in plasma levels of the compound.

[0209] The following examples further illustrate the present invention. These examples are provided for illustrative purposes only and should not be construed as limiting the invention in any way. [Example]

[0210] material and method Chemicals, recombinant proteins, and antibodies: Mouse anti-human p38α antibody and rabbit anti-phosphorylated MK2 (T222) and anti-phosphorylated Stat-1 (S727) antibodies were purchased from Cell Signaling Technology (Danvers, MA). The coding sequences of human p38α variant 2 and p38β (with an N-terminal HA tag) were amplified by PCR and cloned into pRSetA (Thermo Fisher). Mutations were introduced into p38α using Quikchange (Stratagene) and confirmed by bidirectional sequencing. Plasmids were transformed into E. coli BL21, and proteins were purified using a cobalt column (TALON™; Clontech Laboratories; Mountain View, CA) and confirmed by SDS-PAGE and Western blot. Compounds identified in the CADD screen were purchased from Maybridge Chemical Co. (Belgium).

[0211] CADD Identification of Lead Compounds (Figure 1d): Based on the X-ray crystal structure of mouse p38α / MAPK14 (PDB ID: 1P38), a stepwise iterative CADD process was used to screen an in silico database of small molecule compounds available from the Maybridge Chemical Screening Collection for their potential to bind in a pocket near the ED substrate-binding site (Figures 1a and 1b). In silico preparation of p38α conformations was performed using the CHARMM36 and Nanoscale Molecular Dynamics (NAMD) programs (CGenFF) general force fields to identify potential local ligand-binding pockets. Protein structures were subjected to clustering to identify 20 representative protein conformations to account for protein flexibility. The screening process consisted of the following steps: (1) identifying potential inhibitor binding sites; (2) ranking compounds based on their van der Waals (VDW) energy and electrostatic interaction energy with the protein binding pocket using the program DOCK with size-based score normalization; (3) subjecting the top 50,000 compounds to a second round of in silico screening involving further relaxation of the ligand during simulated binding, and selecting the top 1,000 compounds based on their total interaction energy, including ligand size-based score normalization; (4) performing a cluster analysis based on the chemical fingerprints of the top-scoring compounds using the program MOE (Chemical Computing Group) to identify chemically diverse compounds; and selecting a final list of potential p38α interacting compounds based on 4DBA, a scalar bioavailability metric that considers physicochemical descriptors in Lipinski's rule of five.

[0212] Mouse unphosphorylated p38α / MAPK14 variant-1 differs from its human variant-2 by only two amino acids (H48L and A263T), and mouse variant-2 and human variant-1 differ by only 14 amino acids between residues 230 and 254. Neither these amino acid differences nor the phosphorylation state of p38α (Figure 1c) are predicted to significantly alter the structure of the CD or ED sites or our CADD target sites, validating the use of mouse unphosphorylated p38α variant-1 for CADD searches and unphosphorylated recombinant human p38α variant-2 protein for DSF screening.

[0213] Site Identification by Ligand Competition Saturation (SILCS): Using the site identification by ligand competition saturation (SILCS) method (Figure 6, potential binding sites in green), an in silico map of all potential ligand-binding pockets of p38α, including the ED site target, was completed. The SILCS method creates free energy maps (Grid Free Energy; GFE FragMaps) of functional group interaction patterns of p38α, which enable the identification of putative binding sites and rapid free energy estimation of ligand binding to various p38α sites (ligand GFE or LGFE). The SILCS GFE FragMaps consider protein flexibility, protein desolvation, functional group desolvation, and functional group-protein interactions, resulting in highly accurate mapping of the protein for use in database screening and lead optimization. Each step of in silico CADD screening of any compound database begins with the SILCS pharmacophore approach, which takes protein flexibility into account. Secondary screening is based on the MC-SILCS approach, in which relative free energies of binding are calculated. Final screening based on chemical diversity, physiochemical properties maximizing absorption, distribution, metabolism, and excretion (ADME) characteristics, and potential chemical optimization generates a list of compounds to test for selective p38α binding and biological activity. Additional rounds of screening are performed using a modified CADD strategy based on proteomic and structural analysis of lead compounds. Database searches for structural analogs of lead compounds from previous rounds are performed using the program MOE (Chemical Computing Group).

[0214] Alternative CADD methods: The program Dock can be used with scoring based on Dock van der Waals (vdW) interaction energies normalized to molecular weight (MW). This method identifies compounds that sterically fit into the binding site, with a bias toward low MW compounds. Further ranking of compounds uses a generalized linear response method, including the free energy of solvation based on the implicit generalized Born (GB) model.

[0215] Alternative p38α targets: The search strategy can be modified to target the CD or DEF site. Because activation of p38α is required for the formation of the DEF pocket, dual-phosphorylated p38α is used for the DSF screen.

[0216] Differential Scanning Fluorescence (DSF): Binding of CADD-selected compounds to p38α and p38β isoforms was experimentally tested using DSF to assess changes in the melting temperature (ΔTm) of the target protein upon interaction with the test compound. SYPRO Orange (Invitrogen; diluted 1:1000 in 10 mM HEPES, 150 mM NaCl, pH 7.5) and 1 μM unphosphorylated recombinant human p38α were added to a 96-well PCR plate, followed by the addition of 50 nM–200 μM test compound in 100% DMSO (final DMSO concentration of 2%). The plate was mixed, sealed, and centrifuged at 1000 rpm for 1 minute. Melting curves were obtained using an Applied Biosystems real-time PCR instrument. Melting points were determined from the first derivative curves. Additionally, p38β or target-disrupted p38α mutants were also used.

[0217] Although DSF is less sensitive than other assays for ligand:protein binding, it is low-cost and has relatively high throughput. DSF detected p38α binding in 25% of CADD-identified compounds screened, increasing selective p38α binding by 10%, demonstrating the good efficiency of both the CADD and DSF screening strategies. The 10% hit rate of the CADD search for substrate-selective p38α inhibitors was similar to that of searches for substrate-selective ERK inhibitors and much greater than the typical 0.1–0.01% hit rate using experimental screening alone.

[0218] Cell culture: HMVECL were purchased from Promocell (Heidelberg, DE), maintained in endothelial cell growth medium MV2, and used at passages 3–10 and tested after confluence according to the supplier's protocol. The THP1 human monocytic cell line (American Type Culture Collection / ATCC, no. TIB202) was maintained in RPMI 1640 supplemented with 2 mM L-glutamine, 1 mM sodium pyruvate, 10 mM HEPES buffer, pH 7.3, penicillin, streptomycin, 0.05 mM β-mercaptoethanol, and 10% defined fetal bovine serum (FBS; Gibco Life Technologies, Grand Island, NY). HeLa cells (ATCC no. CCL-2) were cultured in DMEM containing 4.5 g / L glucose, 1 mM sodium pyruvate, 2 mM L-glutamine, penicillin, streptomycin, and 10% FBS. Before experiments, THP1 cells were differentiated by treating them with 5 ng / ml phorbol 12-myristate 13-acetate (PMA, Sigma-Aldrich) for 24 hours, washing with PBS, and culturing them in PMA-free medium at 37°C for an additional 24 hours.

[0219] Endothelial permeability assay: The permeability of HMVECL monolayers was assessed by measuring the transendothelial flux of 10 kDa dextran conjugated to Cascade Blue fluorescent dye in Matrigel-coated 3 μm pore size Transwell plates for 30 min at 37°C.

[0220] Cells were treated with 1-100 μM test compound, 10 μM SB203580, or DMSO for 1 hour, followed by treatment with 10 ng / ml rhTNFα for 6 hours at 39.5°C. Permeability was assessed by adding 100 μg / ml Cascade Blue-conjugated 10 kDa dextran to the bottom wells for 30 minutes at 37°C and analyzing the fluorescence (400 / 420 nm) of the top wells.

[0221] Neutrophil transendothelial migration (TEM) assay: Using a protocol approved by the University of Maryland Institutional Review Board, neutrophils were isolated from heparinized venous blood drawn from healthy volunteers, and TEM of calcein-labeled neutrophils was measured via HMVECL.

[0222] The cytotoxicity of HMVECL exposed to 10-100 μM of each compound is analyzed by MTS assay (Promega), LDH release (Promega), and immunoblot for activated caspase-3 (Cell Signaling).

[0223] Macrophage cytokine expression: We evaluated the ability of test compounds to block LPS-induced cytokine expression in PMA-differentiated THP1 cells using qRT-PCR and Luminex-based immunoassays (UMB Cytokine Core Lab). THP1 cells differentiated with 5 ng / ml PMA for 24 hours were treated with 1–100 μM test compound, 10 μM SB203580, or DMSO for 1 hour, followed by treatment with 100 ng / ml ultrapure E. coli 0111:B4 LPS (InvivoGen) for 3 hours (qRT-PCR; Real Time Primers) or 24 hours (supernatants for immunoassay).

[0224] Murine acute lung injury model: Male CD-1 mice weighing 25–30 g were purchased from Charles River and housed at the Baltimore Veterans Administration Medical Center Animal Care Facility under AALAC-approved conditions. All protocols were approved by the University of Maryland, Baltimore, IACUC. Inhibitors were tested via intratracheal administration (i.t.) in mice. LPS / FRH-induced ALI model: Mice were pretreated with SB203580 or a putative p38 inhibitor (in 2% or less DMSO) via intraperitoneal (i.t.) injection 0.5 ml 1 h before intratracheal instillation of 50 μg LPS and switched to a 37°C incubator to raise the core temperature to approximately 39.5°C. Mice were euthanized 24 h later, and lungs were lavaged with a total of 2 ml of PBS. Cells were counted, and the cell-free lavage fluid was analyzed for protein content using the Bradford method (Biorad).

[0225] To implant intraperitoneal thermistors, animals are anesthetized with isoflurane inhalation during surgery. Mice receive 0.05–0.1 mg / kg buprenorphine analgesia (sc) every 12 hours for 2 days postoperatively. Buprenorphine analgesia is administered if significant distress occurs during the ALI model. LPS is administered in 50 μl of PBS via instillation in the posterior oropharynx during isoflurane anesthesia. p38 inhibitors are administered via IP injection with a 25 g needle while the mice are conscious and lightly restrained.

[0226] The combination of FRH and intratracheal LPS induces robust pulmonary neutrophil influx, cytokine expression, and protein leakage by 12–24 h, leading to 50% mortality beginning at 48 h. In this model, UM101 was more potent than SB203580 in reducing neutrophil and protein accumulation in the BAL. Therefore, to minimize the number of mice required for this screening, lung injury, pulmonary and extrapulmonary inflammation, and drug toxicity are measured at a single 24 h time point, including BAL protein, neutrophil, and proinflammatory cytokine content, serum levels of IL-6, creatinine, and AST (Abcam), and cardiac troponin I (MyBiosource). Novel compounds are tested at doses of 4, 12, and 40 mg / kg and compared to vehicle (DMSO) and SB203580 (40 mg / kg)-treated controls. All vehicle- and drug-treated mice are exposed to LPS / FRH (it) and compared to naive mice. Four mice per group may be used.

[0227] Generally, screening is performed in a preventative model and final candidates are evaluated in a therapeutic model.

[0228] Inhibition of substrate phosphorylation: We performed functional analysis of UM101 in blocking p38-dependent phosphorylation of MK2 and Stat-1 in HeLa cells. Cells were pretreated with SB203580 or UM101 for 30 min and then activated with 10 μM anisomycin for 30 min. Cell extracts prepared in RIPA buffer containing protease and phosphatase inhibitors were separated by SDS-PAGE, transferred to PVDF membranes, blocked with 5% dried nonfat milk, and probed with primary antibodies against phosphorylated MK2 and phosphorylated Stat-1, as well as total p38α as a loading control. Bands were detected using secondary antibodies conjugated to infrared fluorophores and infrared fluorescence imaging (Odyssey; LICOR).

[0229] Cytotoxicity Assay: Cytotoxicity was monitored in parallel on monolayers of HMVEC-L formed in 96-well culture plates using a colorimetric assay that measures the reduction of 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) to a formazan dye (CellTiter96™, Promega, Madison, WI) according to the manufacturer's protocol, and quantifies product formation by measuring absorbance at 490 nm.

[0230] Gene expression: RNA integrity was confirmed using an Agilent Bioanalyzer 2100, and all samples were confirmed to have an RNA integrity score (RIN) of 10 before further analysis. Poly(A)-enriched samples were reverse transcribed and sequenced using the Illumina HiSeq platform to generate at least 90 million reads per sample. Intergenic sequences accounted for less than 0.7% of all reads, indicating minimal genomic DNA contamination. Raw data were analyzed using the TopHat read alignment tool and the Homo sapiens genome reference sequence (Ensembl version GRCh38.78). Differential gene expression was analyzed using the DESeq R package (Bioconductor) and a negative binomial distribution model. The criteria for significant differences in gene expression were: (1) a false discovery rate (FDR) of less than 0.05, (2) expression levels greater than the 10th percentile, and (3) a fold change of 2-fold or greater. Differential gene expression patterns were further analyzed using PathwayNet (Troyanskaya Lab, Princeton) and Ingenuity™ Pathway Analysis (Qiagen) tools. Cytokine gene expression in THP1 cells was analyzed by quantitative RT-PCR using primers and SYBR-green reaction mixtures (Biorad) from a commercially available PCR array (HCA-II Array; Real Time Primers; Elkins Park, PA) and a BioRad iCycler IQ optical module, according to the supplier's protocol. Data were quantified using the Ct difference method of gene expression and normalized to the level of the housekeeping gene, GAPDH, using Ct values ​​automatically determined by the thermocycler.

[0231] Saturation Transfer Difference Nuclear Magnetic Resonance (STD-NMR): A 40 mM stock solution of UM101 was prepared in d6-DMSO. The STD-NMR sample contained 150 mM NaCl, 50 mM phosphate (pH 7), 200 μM UM101, and 5 μM p38 protein in DO. Spectra were recorded at 300 K on an Agilent DD2 500-MHz spectrometer equipped with a 5 mm inverted HFCN probehead. During each transient, the protein was saturated using a series of 58 Gaussian-shaped pulses (50 ms, with a 1 ms delay between pulses) for a total saturation time of 3 s using the vendor-supplied STD-ES pulse sequence. Resonant irradiation of the protein was performed at 0.5 ppm, and non-resonant irradiation was performed at 30 ppm. The vendor-supplied WATERGATE pulse sequence was used to suppress the water signal in the STD spectrum. Resonant and non-resonant pulse sequences were internally subtracted. A total of 16,384 transients were collected for each STD experiment with a 1-second delay between collections, a spectral width of 6000 Hz, and an acquisition time of 1.3 seconds.

[0232] Comparative proteome and phosphopeptide expression profiling by mass spectrometry (MS): Protein expression and the percentage of phosphorylation of specific proteins are quantified label-free using mass spectrometry-based techniques. Specifically, LC-MS / MS is used to compare the effects of compounds of the present invention on protein phosphorylation patterns and proteome expression in TNFα-stimulated HMVECL cells and LPS-stimulated THP1 cells with those of SB203580. Cells were treated with 10 μM SB203580 or EC 50 and E.C. 90Cells are pretreated with test compounds (based on HMVECL permeability and THP1 IL-8 expression assays) for 30 min. For phosphopeptide analysis, cells are stimulated for 0.5, 1.5, and 4 h. Tryptic phosphopeptides are enriched using the commercially available Tio2 enrichment protocol (Pierce) and then analyzed on a nanoUPLC-coupled Thermo Orbitrap Fusion Tribrid mass spectrometer using three strategies: (1) hybrid electron transfer (ETD) / higher-energy collisional (HCD) dissociation (EThcD), (2) data-dependent decision tree (DDDT) logic, (3) HCD product-dependent ETD (HCD-pd-ETD), and / or (4) ion mobility-coupled parallel mass spectrometry (UDMS). e For comparative proteome expression analysis, cells were stimulated for 4 and 12 hours and lysates were analyzed by UDMS. E and ADAPT-DDA on a nanoUPLC-coupled Waters Synapt G2S and / or nanoUPLC-coupled Thermo Orbitrap Fusion Tribrid. Relative peptide abundances were measured by comparing MS1 peak areas of peptide ions and analyzed using the different fragmentation strategies described above (EThcD, DDDT, HCD-pd-ETD, and UDMS). e ) is used to confirm its identity and phosphorylation event by MS2 sequencing. The Aligned AMRT (Accurate Mass and Retention Time) cluster quantification algorithm as described is used for label-free quantification.

[0233] Immunoblot analysis: Changes in the total in vivo proteome and phosphoproteome are confirmed by immunoblotting using commercial antibodies and infrared fluorescence imaging (Odyssey; LICOR). In vitro kinase assays are performed in reactions containing recombinant active p38α and one or more recombinant substrate proteins and analyzed by immunoblotting with phosphorylation-site-specific antibodies.

[0234] X-ray crystallography: X-ray crystallography provides high-resolution analysis of compound binding to p38α. The primary approach for growing p38α involves cocrystallization of compound crystals at a 2:1 compound:p38α molar ratio. Alternatively, the compound is soaked in preformed p38α crystals. Diffraction-quality protein crystals are grown and screened using an automated system including an Alchemist DT screenmaker, a Gryphon drop setter with an LCP module, and a Minstrel DT UV / Vis automated visualization system (Rigaku). These structures are solved by molecular replacement using known p38α structures and standard crystallography software (SBGrid).

[0235] Analysis of p38 binding kinetics: The KD of compounds of the present invention is estimated by DSF. ITC is performed to generate thermodynamic information to refine compound KD calculations and facilitate ligand optimization. Data are collected using an Auto ITC HT microcalorimeter (MicroCal). Recombinant p38α (10 μM) and a stock concentration of test compound (200 μM) are prepared and degassed in the same buffer containing low ionization energy (e.g., 50 mM phosphate or 50 mM NaCl in citrate). Exotherms / endotherms during compound titration are measured and analyzed using MicroCal software.

[0236] Pharmacokinetic / pharmacodynamic (PK / PD) analysis of lead compounds: Compounds are comprehensively analyzed for in vivo toxicity and efficacy as both preventive and therapeutic agents in an intratracheal LPS + FRH-induced mouse ALI model. This model is a short-term model of human ARDS, amenable to parenteral administration of therapeutic agents, and characterized by extensive endothelial permeability, neutrophil accumulation, proinflammatory cytokine and chemokine expression, epithelial damage, and approximately 50% mortality beginning at 48 hours. Results are generalizable to other inflammatory diseases. Compounds are solubilized in DMSO to a final concentration of 1% or less and administered as a single intraperitoneal injection. The maximum tolerated dose (MTD) is determined by monitoring mice for signs of distress (including changes in motor activity, weight loss, reduced grooming, and ruffled hair), creatinine, BUN, aspartate transaminase (AST), and cardiac troponin levels for 24 hours. Inhibitors are administered either 30 minutes before or 8 hours after LPS+FRH as prophylactic or therapeutic models, respectively.

[0237] Animal number and gender: All studies were performed in CD1 mice, a robust strain with validated ALI and pneumonia models. Dose escalation follows published guidelines, using two mice per dose and observation for 24 hours. Differences in survival rates are tested in groups of 20 mice (detecting a survival difference of 75% vs. 25%, α=0.05, β=0.2). Group sizes of six mice are used for BAL and plasma analysis of injury / inflammation, as well as lung homogenates for analysis of apoptosis signaling, and four mice per group are used for histology. Survival experiments are performed with equal numbers of male and female mice, and differences are compared by two-way ANOVA. Additional experiments are performed to analyze any unexpected gender differences discovered in drug effects. In some embodiments, male mice are used in the experiments.

[0238] The maximum tolerated dose (MTD) of the most potent structurally distinct compound is determined by measuring toxicity at 20, 40, and 80 mg / kg (ip) in two mice per dose, monitored for 24 hours, and euthanized. Serum is analyzed for markers of liver, kidney, and heart toxicity. Kidneys, heart, liver, and lungs (inflated) are fixed, paraffin-embedded, stained with hematoxylin and eosin, and examined for inflammation and damage. Control mice receive vehicle (1% DMSO). The toxic compound is replaced by the next structurally distinct compound on the candidate list.

[0239] Activity of inhibitors to block FRH-enhanced LPS-induced ALI: Compounds are tested at the MTD in the LPS- and FRH-induced ALI model.

[0240] Effect of pretreatment on survival: In LPS / FRH-challenged mice (groups of 20 mice), the efficacy of pretreatment with test compound at the MTD on survival is compared with SB203580 at 40 mg / kg and vehicle (1% DMSO). Mice receive pretreatment as a single 0.5 ml injection and, 30 minutes later, receive 50 μg of LPS via it instillation and placed at an ambient temperature of 37°C. This exposure increases core temperature from 36.5°C to 39.5°C, which is confirmed in some mice using telemetric temperature monitoring (Data Sciences International; St. Paul, MN). Mice are monitored for survival using moribundity instead of death. Compounds that demonstrate a survival advantage over DMSO are further analyzed for efficacy when administered 24 hours after LPS. Ineffective compounds are replaced with the next compound on the candidate list. Effective compounds are further tested at 10% and 30% of the MTD.

[0241] Post-LPS effect on survival: Compounds effective as pretreatments will be analyzed for efficacy at the same dose using the same protocol, except that the LPS instillation and initiation of FRH will be delayed until 8 hours. Compounds that confer a survival advantage over SB203580 will be analyzed for biological effect and pharmacokinetics. Ineffective compounds will be replaced with the next compound on the list.

[0242] Compound Effects on Inflammation, Lung Injury, and Permeability: The most effective compounds in survival experiments will be further analyzed for their effects on lung injury and inflammation in the LPS+FRH ALI model. Mice were placed in a 1000 mL LPS / FRH incubator at 30 min before or 8 h after LPS / FRH challenge. 50 Mice were pretreated with each compound (based on survival experiments), 40 mg / kg SB203580, or DMSO and euthanized 24 hours after LPS. Six mice per group were analyzed for neutrophil content by counting modified Giemsa-stained cytopreps, total protein by the Bradford method, and cytokine levels by Luminex-based immunoassays (UMB Cytokine Core Lab). After lavage, lungs were excised, snap-frozen in liquid nitrogen, and homogenates prepared for immunoblotting of candidate p38α substrates to confirm the substrate inhibitor effects observed in vitro. Lungs from four mice per group were flattened and fixed at 20 cm H2O, Prefer™, paraffin-embedded, and stained with hematoxylin and eosin or immunostained with GR-1 to assess lung injury and neutrophil infiltration, as well as TUNEL staining and immunostaining for active caspase-3 to assess apoptosis. Serum IL-6 is measured as an index of systemic inflammation.

[0243] Pharmacokinetics of novel p38 modulators: Characterize the pharmacokinetics (PK) of active compounds in mice. First, develop bioanalytical methods for each compound and validate them according to FDA guidance. Then, perform PK studies to characterize pulmonary uptake and key PK parameters (i.e., clearance (CL), volume of distribution (Vd), and maximum plasma concentration (C)) that characterize each compound. max ), C maxArrival time (T max ), area under the plasma concentration curve (AUC) and half-life (t 1 / 2 ) are determined. PK parameters are used to estimate the time required to reach steady-state plasma concentrations (corresponding to 5 half-lives) and provide guidance for dose selection for further PD studies. Additionally, these studies help rank the tested p38 modulators in terms of lung / plasma concentration ratios. For each study, CD1 mice (n = 30) are treated with a single ip dose (10-50 mg / kg) of the selected p38 modulator (the dose range for each p38 modulator will depend on the results of the studies outlined above). In some embodiments, mice (n = 3 / time point) are euthanized pre-dose and at 5, 15, 30, 60, 120, 240, 360, 600, and 720 minutes post-dose. Blood and lung samples are analyzed using validated HPLC methods.

[0244] Data Analysis: Pathways modified by compounds of the present invention compared with SB203580 are predicted by: (1) analyzing comparative proteome expression using Ingenuity pathway analysis and PathwayNet, as well as RNASeq data from UM101, and (2) analyzing comparative phosphoproteomes using quantitative approaches and bioinformatics. Mass spectrometry results are confirmed by analyzing the phosphorylation of candidate substrates in cells and by in vitro kinase assays with immunoblots. Off-target binding suggested by proteomic data is confirmed by evaluating test compounds over a wide concentration range and in vitro kinase reactions using DSF and STD-NMR, as well as phosphoimmunoblots for specific substrates. By identifying common pathways modified by multiple lead compounds and their interactions with p38α, the common p38α effects required for their favorable biological activity are predicted and incorporated into the CADD algorithm for subsequent search and lead optimization.

[0245] Since the objective of this invention is, in one embodiment, to identify and characterize the PD / PK properties of novel anti-inflammatory compounds, these compounds are, in one embodiment, tested in order of activity based on functional screening, and compounds that fail toxicity or efficacy in survival studies are replaced with the next most potent structurally distinct compound. Compounds are compared to vehicle alone and SB203580 using one-way ANOVA / Fisher's PLSD. PK data are analyzed by the naive mean method. Compartmental modeling is used to estimate various pharmacokinetic parameters using the Phoenix platform (ver. 1.3, Pharsight, Sunnyvale, CA). Several compartmental models are evaluated to determine the best-fit model. Equal weighting, 1 / y, 1 / y^, 1 / y 2 , and 1 / y^ 2 Various weighting schemes are used, including: (where y is the observed drug concentration and y^ is the model-predicted drug concentration). In some embodiments, the final model is selected based on goodness-of-fit plots, weighted sum of squared residuals, random distribution of residuals, precision of parameter estimates, Akaike's Information Criterion, and Schwarz's Criterion. After the final model is developed, plasma CL, Vd, C max , T max , AUC, and t 1 / 2 Estimates of PK parameters are reported, including: Lung uptake is expressed as the lung / plasma (L / P) concentration ratio.

[0246] Alternative approach: If phosphorylation site-specific antibodies are unavailable and phosphorylation does not cause a detectable shift on immunoblots, cell lysates can be enriched using TiO2 prior to immunoblotting. The incubation time can be adjusted as needed based on the results of in vitro and in vivo proteomics and immunoblotting. Despite the use of maximal starting material or isolated cell fractions, low protein abundance can prevent the detection of phosphorylated proteins in cell lysates. In this case, LC-MS-MS can be used to enhance in vivo cellular phosphoproteome analysis and comprehensively analyze the effects of inhibitors on phosphopeptide patterns in an in vitro kinase assay of p38α using cell lysates as substrates after inactivating the endogenous kinase with 5'-4-fluorosulfonylbenzoyl adenosine (FSBA). If results without labeling are equivocal, stable isotope dimethyl labeling can be used. Other supporting techniques include hydrogen-deuterium exchange mass spectrometry and NMR, as well as DSF / STD-NMR to assess binding to wild-type p38α and CADD target mutants. Surface plasmon resonance (SPR) (Biacore T200 Core) can be evaluated as an alternative to ITC to reduce protein / compound requirements.

[0247] Statistical methods: Data are presented as mean ± SE. Differences between more than two groups were analyzed by applying Tukey's honestly significant difference test to one-way analysis of variance (ANOVA). Differences between dose-response curves were analyzed by multivariate ANOVA (MANOVA), with a p-value of less than 0.05 considered significant.

[0248] Example 1: CADD modeling of p38 MAPK substrate docking site, compound identification, and screening of compounds for direct, selective interaction with p38α The inhibitors and methods of the present invention involve a CADD-based strategy to identify low molecular weight compounds predicted to bind near the ED substrate docking site of mouse unphosphorylated p38α (MAPK14 variant-1; PDB: 1P38), which is over 99% identical to human p38α (variant-2) (Figure 1a). The ED and CD sites of p38α are located at either end of the substrate-binding groove, located opposite the catalytic site of the protein (Figure 1a). A pocket near the ED binding site containing 10 amino acids was identified, only 7 of which are identical between p38α and p38β (Figure 1b). Structural superposition of mouse unphosphorylated (PDB: 1p38) and dually phosphorylated p38α (PDB: 3PY3) revealed that the target pockets of the two forms overlap almost perfectly (Figure 1c).

[0249] An overview of the CADD screening and compound testing protocol is shown in Figure 1d. Compounds from the Maybridge screening collection were analyzed for binding to the target p38α pocket based on van der Waals (VDW) and electrostatic interaction energies, chemical diversity via chemical fingerprint-based cluster analysis, solubility, molecular weight, and the number of hydrogen-bonding functional groups that maximize bioavailability.

[0250] From a panel of 150 diverse compounds (Table 3) selected for potential biological testing, 20 structurally distinct compounds were selected for functional analysis (Table 2). [Table 4] [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4]

[0251] Using DSF, test compounds were screened at 10–100 μM for binding to recombinant p38α and ERK2 (Figure 1e, Table 1). Five compounds induced concentration-dependent stabilization of p38α and demonstrated binding. Three of these compounds also stabilized ERK2 (3, 5, and 141, highlighted in yellow (denoted with a "*")), while two, namely, UM60 (N2,N7-di(2-hydroxyethyl)-9-oxo-9H-2,7-fluorenedisulfonamide) and UM101 (4-chloro-N-{4-[(1,1-dioxo-1lambda-6,4-thiazin-4-yl)methyl]phenyl}benzamide), stabilized p38α but not ERK2 (highlighted in blue (denoted with a "+")). These two structurally distinct compounds (Fig. 1f), added at 100 μM, increased the melting temperature of p38α by approximately 0.7°C, compared with the 6°C increase caused by SB203580.

[0252] MC-SILCS docking of UM101 to the ED site and GFE FragMap analysis identified several structural features that could be modified to improve selectivity and potency (Figure 7). The modifiable sites on UM101 correspond to the sites identified as interacting with p38α in the NMR STD analysis (Figures 3f-3k).

[0253] Example 2: Effect of compounds on endothelial barrier function We tested the ability of UM60 and UM101 to stabilize the endothelial barrier to macromolecules and neutrophils in TNFα- and heat-challenged HMVECL monolayers (Figure 2). A 6-hour exposure to 1 ng / ml TNFα combined with heat (39.5°C) increased permeability to 10 kDa dextran by 2.8-fold compared to untreated cells at 37°C. Pretreatment with 10 μM SB203580 for 30 minutes reduced TNFα / heat-induced permeability by 50% (Figure 2a). Pretreatment with 10 and 25 μM UM60 had no effect on permeability, but 100 μM UM60 reduced the TNFα / hyperthermia-induced increase in permeability by 71%, whereas 10, 25, and 100 μM UM101 reduced the TNFα / hyperthermia-induced increase in permeability by 74%, 89%, and over 100%, respectively.

[0254] Preincubation of HMVECL at 39.5°C for 6 hours resulted in a subsequent transendothelial migration (TEM) of IL-8-specific neutrophils of 22.8 ± 0.45 × 10 3 to 31.8±0.54×10 3 (neutrophil count) (Figure 2b). Pretreatment with 10 μM SB203580 reduced hyperthermia-enhanced neutrophil TEM by 84%. UM60 at 10 and 25 μM and UM101 at 10 μM reduced the hyperthermia-enhanced increase in TEM by 18%, 89%, and 95%. UM60 at 50 μM and UM101 at 25 and 50 μM reduced TEM below baseline levels. When added to HMVECLs at 100 μM for 48 h, neither compound was toxic in the LDH release and MTS assays.

[0255] Example 3: Comparison of the efficacy of SB203580 and UM101 in mouse ALI We compared the efficacy of UM60, UM101, and SB203580 in mitigating transalveolar protein and neutrophil extravasation in a mouse model of LPS / hyperthermia-induced ALI (Figures 2c and 2d). Mice received a single intraperitoneal injection of 100, 300, 500, or 1000 μg of UM101, 1000 μg of UM60, or 1000 μg of SB203580 in 0.5 ml of 2% DMSO 30 min before intratracheal instillation of 50 μg of LPS and were then transferred to a hyperthermic chamber. Control mice received DMSO. Four of six UM60-treated mice, one of six SB203580-treated mice, and one of 11 DMSO-treated control mice died within 24 h. All 16 UM101-pretreated mice survived. Lung lavage fluid from DMSO-pretreated, LPS / heat-challenged mice contained 1.09 ± 0.19 mg / ml protein and 3.97 ± 1.07 × 10 6 Compared to DMSO-treated controls, lavage protein concentration and neutrophil content in mice pretreated with 1000 μg of SB203580 were reduced by 42% and 46.8%, respectively. Lavage protein concentration in mice pretreated with 100 μg, 300 μg, 500 μg, and 1000 μg of UM101 was reduced by 0%, 44.1%, 43.9%, and 92.9%, respectively, and lavage neutrophil content was reduced by 44.4%, 49.5%, 55.3%, and 54%, respectively.

[0256] Example 4: Effects of SB203580 and UM101 on LPS-induced gene expression in human THP1 promonocytes The effects of UM101 and SB203580 on inflammatory cytokine expression were compared by pretreating PMA-differentiated THP1 cells with 25 μM SB203580 or 10, 25, or 100 μM UM101 for 30 minutes, then stimulating them with 100 ng / ml LPS. After 4 hours, RNA was harvested and analyzed by PCR-based cytokine array analysis. SB203580 inhibited the expression of 7 of the 16 LPS-stimulated genes in the array: IL-1α, IL-8, TNFSF8 (CD30 ligand), TNFSF9 (CD137 ligand), CXCL5, CCL7, and CCL17 (Table 4). UM101 inhibited the expression of all SB203580-inhibited genes except TNFSF9, and also inhibited the four SB203580-insensitive genes: IL-1β, CXCL1, TNFSF15, and CCL5. [Table 6]

[0257] Example 5: Comparison of the effects of SB203580 and UM101 on TNFα-induced gene expression in HMVECL We compared the effects of UM101 and SB203580 on TNFα-induced gene expression in HMVECLs using RNASeq. HMVECLs were pretreated with 10 μM SB203580 or 100 μM UM101 for 1 hour, followed by stimulation with 10 ng / ml TNFα for 3 hours. A UM101 concentration 10-fold higher than its biologically effective dose in the HMVECL barrier assay was used to ensure identification of any partial overlap with SB203580. The TNFα concentration and duration of stimulation used were based on previously published studies and confirmed by preliminary qRT-PCR analysis of IL-8 and IL-1β mRNA expression (Figure 4). After filtering the RNASeq results for genes with 10 or more reads in at least one sample per experiment, we found 511 genes that were upregulated by more than two-fold and 520 genes that were downregulated by more than two-fold by TNFα treatment (Table 5). Table 7-1 Table 7-2 Table 7-3 Table 7-4 Table 7-5 Table 7-6 Table 7-7 Table 7-8 Table 7-9 Table 7-10 Table 7-11 Table 7-12 Table 7-13 Table 7-14 Table 7-15 Table 7-16 Table 7-17 Table 7-18 Table 7-19 Table 7-20 Table 7-21 Table 7-22 Table 7-23

[0258] SB203580 inhibited the expression of 61 TNFα-induced genes, 28 of which were also inhibited by UM101 (Table 6, Table 5, Figure 5). SB203580 increased the expression of 38 genes, 10 of which were also increased by UM101. Of the 28 genes inhibited by both SB203580 and UM101, 22 encoded known proteins, including IL-1β, CCL17, MMP9, IDO1, CXCL5, CXCL10, and CXCL11, hyaluronan synthase-3, MUC4, and PLA2 (Table 6). Of the 33 genes inhibited by SB203580 but not by UM101, 24 encoded known proteins and included GM-CSF, IL-1α, TNFα, IL-12 receptor-β1, and hyaluronan synthase-2 (Table 6 ). [Table 8-1] [Table 8-2]

[0259] The differentially expressed genes were further analyzed using PathwayNet and Ingenuity™ tools to identify transcription factors and biological pathways regulated by the two inhibitors. PathwayNet analysis suggested that UM101 inhibits some transcription factors inhibited by SB203580 (Stat-1, c-Fos, c-Jun, NFκB, p53, PPARγ, and Sp1), but not others (ATF1, ATF2, Elk1, c / EBPβ, USF1, SMAD3, FOXO1, and CREB via MSK1 / 2). Ingenuity™ analysis suggested that both SB203580 and UM101 inhibited dendritic cell maturation, triggering receptor expressed on myeloid cells-1 (TREM1), high-mobility group box 1 (HMGB1), and NFκB pathways, and both increased liver X receptor / retinoid X receptor (LXR / RXR) activation, whereas only SB203580 inhibited IL-6, acute-phase, and cholecystokinin / gastrin-mediated pathways (Figure 3a). UM101 at 100 μM reduced the expression of 115 genes and increased the expression of 119 genes that were not modified by SB203580 (Table 5). Ingenuity™ pathway analysis suggested reduced Toll-like receptor and Wnt / β-catenin signaling, and increased nitric oxide in cardiovascular disease pathways (Figure 3b).

[0260] Example 6: Comparison of the effects of SB203580 and UM101 on p38 MAPK substrate phosphorylation profiles To assess whether UM101 selectively inhibits phosphorylation of its target, HeLa cells were pretreated with 10 μM SB203580, 50 μM UM101, or 0.1% DMSO (vehicle control) for 30 min and then treated with the p38 activator anisomycin (25 μg / ml). Phosphorylated MK2 and Stat-1 were analyzed by immunoblotting (Fig. 3c). Anisomycin-stimulated phosphorylation of MK2 and Stat-1 was reduced by both 10 μM SB203580 and 50 μM UM101, but was more reduced by SB203580.

[0261] Example 7: Analysis of specific binding of UM101 to p38α DSF was used to analyze the concentration-specific binding of UM101 to p38α and p38β. SB203580 stabilized both p38α and p38β, whereas UM101 stabilized only p38α (Figure 3d). To confirm that UM101 bound to the CADD target pocket, DSF was used to compare the binding of UM101 and SB203580 to wild-type p38α and mutant p38α (R49K / HL107-8TF / K165R, in which four of the ten target pocket amino acids were substituted) (Figure 3e). The mutant showed binding to SB203580, identical to that of wild-type p38α, but not to UM101.

[0262] Saturation transfer difference (STD)-NMR was used to confirm the selective binding of UM101 to the CADD target pocket of p38α. The 1D spectrum of UM101 in the presence of p38α is shown in Figure 3f, and the STD spectrum of the same sample is shown in Figure 3g. Peaks in the 1D spectrum are labeled according to tentative peak assignments for UM101 in aqueous form, based on the assignments for UM101 in 2 mM d6-DMSO obtained from the use of 1D proton and C13 and 2D-HMBC experiments. The shifts in the peaks in the STD spectrum correspond well with those in the 1D spectrum, thus indicating that protons on both aromatic rings of UM101 interact with p38α. In contrast, although the 1D spectra of UM101 with p38β and mutant p38α were similar to those of UM101 / p38α (Figures 3h and 3j), the interaction of UM101 with p38β and mutant p38α was much weaker, as indicated by the barely discernible aromatic proton peaks in the STD spectra of UM101 with p38β (Figure 3i) and mutant p38α (Figure 3k).

[0263] Example 8: Synthetic methods for preparing exemplary compounds of the present invention General Chemistry Methods: All air- or moisture-sensitive reactions were carried out under a positive pressure of nitrogen using oven-dried glassware. Chemical reagents and anhydrous solvents were obtained from commercial sources and used as received.

[0264] The p38α MAPK inhibitor of the present invention can be prepared by methods generally known in the art. For example, compound UM101 can be prepared as shown in Scheme 1. UM101 can be prepared in two steps from three commercially available fragments (Scheme 1), which facilitates its optimization. Acylation of 4-aminobenzaldehyde with 4-chlorobenzoyl chloride in the presence of diisopropylamine (DIPEA) produces an intermediate aldehyde. Subsequent reductive amination of the aldehyde with thiomorpholine 1,1-dioxide and sodium triacetoxyborohydride (NaBH(OAc)3) gives UM101. [ka]

[0265] Structure-activity relationships (SAR) focused on UM101 and additional lead compounds were performed to determine the pharmacophore and the information used to achieve its optimization, for example, by feeding this information back into the CADD model to improve its predictability and facilitate subsequent design cycles. An overview of proposed modifications to UM101 is shown in Scheme 2. This was driven by SILCS molecular modeling and addresses improved binding affinity and specificity, as well as enhanced physicochemical properties. Importantly, STD-NMR analysis of UM101 confirmed that both of its aromatic rings interact with proteins, and therefore modifications to these rings will affect binding affinity. First, the positively charged (under physiological conditions) piperidine nitrogen of UM101 is retained in some embodiments because it is predicted to interact with negatively charged residues such as D112 and D168. According to FragMaps, aliphatic (e.g., cyclohexyl) or aromatic (e.g., furan) substituents away from the central phenyl ring should enhance binding to proteins through interactions with V30, V38, A51, I84, L108, and L167. Additionally, the presence of a hydrogen bond acceptor map overlapping with the aliphatic map suggests that, in some embodiments, a combined aliphatic / hydrogen bond acceptor group, such as OEt, is incorporated at the ortho position relative to the aniline nitrogen. Hydrogen bond donor groups (e.g., NH2, OH) and / or acceptor groups (e.g., OMe, isoxazole) are incorporated at the ortho and meta positions of the peripheral chlorophenyl ring in some embodiments, and in other embodiments, may replace the chloropyridyl ring. These changes further increase the solubility of the compound. In some embodiments, the chlorine on the chlorophenyl ring interacts moderately with proteins, as judged by SILCS GFE analysis, but this site is altered with alternative hydrophobic and more polar groups in other embodiments. The trans amide bond is modified in some embodiments to a rigid E-alkene, and in other embodiments to a more flexible sulfonamide.Based on SILCS GFE analysis, the sulfone SO group contributes -0.5 kcal / mol to binding, indicating that in some embodiments, this region of the molecule can be utilized to optimize the physicochemical properties of the molecule without compromising binding affinity. For example, the SO group is replaced with a polar oxygen atom in some embodiments, and with an NMe group in other embodiments. [ka]

[0266] Example 9: Novel non-ATP / catalytic site p38 mitogen-activated protein kinase inhibitors Using computer-aided drug design (CADD), we identified novel small molecule compounds that target selected substrate-binding sites and inhibit p38 mitogen-activated protein (MAP) kinase activity, which contributes to acute lung injury. These compounds are unique in that they are non-ATP / catalytic kinase inhibitors, offering an alternative to current p38 MAP kinase inhibitors that target the ATP-binding or catalytic site and block all enzymatic activity. Compound UM101 targets the p38α MAP kinase isoform, which is the major p38 MAP kinase isoform involved in tissue damage associated with chronic inflammation and acute lung injury. Current p38 MAP kinase inhibitors target the ATP-binding or catalytic site, block all enzymatic activity, and lack isoform selectivity. In contrast, the compounds disclosed herein are unique in that their target site is directed against protein-substrate interactions, specifically blocking p38α MAP kinase function.

[0267] Novel UM101 analogs were synthesized (Schemes 3-5) and tested for their interaction with p38α MAP kinase and biological activity. [ka] [ka] [ka]

[0268] Two lines of evidence suggest that this lead compound may improve lung barrier function and prevent lung injury. First, this compound can block thrombin-induced cell permeability in human vascular endothelial cells (Figure 8). Second, in mice challenged with LPS to induce pulmonary inflammation, protein leakage into bronchoalveolar lavage fluid was reduced in the presence of the parent compound UM101 and analogs SF-6-222 and SF-7-009 (Figure 9), or analogs SF-6-222, SF-7-009, and SF-7-044 (Figure 10). As shown in Figure 9, UM101 and analogs inhibit acute lung injury in a mouse model. Mice were pretreated with 1 mg of the test compound (ip) and then treated with 100 mg of LPS (it) at 39°C to induce inflammation. Lung injury was assessed by measuring total protein in the lung lavage fluid. As shown in Figure 10, UM101 and analogs inhibit acute lung injury in a mouse model. All mice received 50 μg of LPS (it) and were maintained at 39°C for 24 hours. All treatments were 1 mg in 0.5 ml of DMSO and were administered 6 hours after LPS. Mean ± SM, n = 5. *, p = 0.03 (vs. DMSO #1), †, p = 0.16 (vs. DMSO #2). Lung injury was assessed by measuring total protein in lung lavage fluid.

[0269] A useful application of the present invention is to provide new therapies for acute lung injury and other diseases caused by inflammation and endothelial permeability. Furthermore, the present invention aims to overcome problems related to toxicity and poor efficacy of current p38 MAP kinase inhibitors used in the clinic to treat inflammation.

[0270] Several patent and non-patent publications are cited herein in order to describe the state of the art related to the present invention, the entire disclosure of each of which is incorporated herein by reference.

[0271] While specific embodiments of the present invention have been described and / or exemplified above, various other embodiments will be apparent to those skilled in the art from the foregoing disclosure. Accordingly, the present invention is not limited to the specific embodiments described and / or exemplified, but is capable of considerable variation and modification without departing from the scope and spirit of the appended claims.

[0272] Furthermore, as used herein, the term "about" means that amounts, sizes, formulations, parameters, shapes, and other quantities and characteristics need not be exact but may be approximate and / or larger or smaller, as desired, to reflect tolerances, conversion factors, rounding, measurement error, etc., as well as other factors known to those of ordinary skill in the art. In general, an amount, size, formulation, parameter, shape, or other quantity or characteristic is "about" or "approximately" whether or not expressly stated as such.

[0273] Furthermore, the transitional terms "comprising," "consisting essentially of," and "consisting of," when used in the accompanying claims in their original and amended forms, define the scope of the claim in terms of what additional, unrecited claim elements or steps, if any, are excluded from the claim(s). The term "comprising" is intended to be inclusive or open-ended and does not exclude any additional, unrecited elements, methods, steps, or materials. The term "consisting of" excludes any elements, steps, or materials other than those specified in the claim, and in the latter case, excludes impurities normally associated with the specified material(s). The term "consisting essentially of" limits the scope of the claim to certain elements, steps, or materials that do not materially affect the basic and novel characteristic(s) of the claimed invention. All compounds, compositions, formulations, and methods described herein that embody the present invention may, in alternative embodiments, be more specifically defined by any of the transitional terms "comprising," "consisting essentially of," and "consisting of." References JPEG2023106403000078.jpg213155 JPEG2023106403000079.jpg214155 JPEG2023106403000080.jpg214155 JPEG2023106403000081.jpg215155 JPEG2023106403000082.jpg215155 JPEG2023106403000083.jpg214155 JPEG2023106403000084.jpg216155 JPEG2023106403000085.jpg215155 JPEG2023106403000086.jpg222155 JPEG2023106403000087.jpg222155 JPEG2023106403000088.jpg139155

Claims

1. A compound of formula I, or a pharma- ceutically acceptable salt thereof: 【Chemistry 1】 [In formula I, L １ is -CH ２ -, -C(CH ３ ) ２ -, and -C(CH ２ CH ２ ) - is selected from L ２ is -NHSO ２ --NHCH ２ --, --CH ２ NH-, -NHCO-, -CONH-, and -SO ２ NH-, -NR 1 R 2 is 【Chemistry 2】 (In the formula, R 7a is selected from hydrogen, OH, C 1-10 alkyl, and substituted C 1-10 alkyl. is selected from R ３ , R ４ , R ５ , and R ６ each is hydrogen; Ar １ teeth, 【Chemistry 3】 (In the formula, R ７ , R ８ , R ９ , R １０ , R １１ , R １２ , and R １３ Each of is independently hydrogen, -NR 1 R 2 , or -NR １a R ２a and R 1a and R 2a are independently halogen, C 1-10 alkyl, or substituted C 1-10 alkyl. . . . is selected.

2. L １ is -CH ２ 2. The compound of claim 1, wherein: -, or a pharma- ceutically acceptable salt thereof.

3. L ２ is -NHSO ２ 3. The compound according to claim 1 or 2, wherein: -, or a pharma- ceutically acceptable salt thereof.

4. The compound according to claim 1 or 2, or a pharma- ceutically acceptable salt thereof, wherein L2 is -NHCH2-.

5. The compound according to claim 1 or 2, or a pharma- ceutically acceptable salt thereof, wherein L2 is -NHCO-.

6. R １a , and R ２a Each of is independently hydrogen, and C １－３ 6. The compound of any one of claims 1 to 5, or a pharma- ceutically acceptable salt thereof, wherein: R is selected from the group consisting of aryl, ... and aryl;

7. R １a , and R ２a or a pharma- ceutically acceptable salt thereof. The compound of any one of claims 1 to 5, wherein each of is independently methyl.

8. Ar １ teeth, 【Chemistry 5】 8. The compound according to any one of claims 1 to 7, which has the structure: or a pharma- ceutically acceptable salt thereof.

9. The following formula 【Chemistry 6】 2. The compound of claim 1, which is compound 1049 shown in Figure 1, or a pharma- ceutically acceptable salt thereof.

10. A pharmaceutical composition comprising a compound according to any one of claims 1 to 9, or a pharma- ceutically acceptable salt thereof.

11. The pharmaceutical composition of claim 10, which is for oral administration.

12. The pharmaceutical composition according to claim 10 or 11 for treating cancer.

13. 12. The pharmaceutical composition according to claim 10 or 11, for treating an inflammatory disease selected from the group consisting of rheumatoid arthritis, cardiovascular disease, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), asthma, acute respiratory distress syndrome (ARDS), and acute lung injury (ALI).

14. Use of a compound according to any one of claims 1 to 9 or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition according to claim 10 or 11, for the preparation of a medicament for treating cancer.

15. Use of a compound according to any one of claims 1 to 9 or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition according to claim 10 or 11, for the preparation of a medicament for treating an inflammatory disease selected from the group consisting of rheumatoid arthritis, cardiovascular disease, multiple sclerosis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), asthma, acute respiratory distress syndrome (ARDS), and acute lung injury (ALI).

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