RESUMEN
We have applied simulated annealing of chemical potential (SACP) to a diverse set of â¼150 very small molecules to provide insights into new interactions in the binding pocket of human renin, a historically difficult target for which to find low molecular weight (MW) inhibitors with good bioavailability. In one of its many uses in drug discovery, SACP provides an efficient, thermodynamically principled method of ranking chemotype replacements for scaffold hopping and manipulating physicochemical characteristics for drug development. We introduce the use of Constrained Fragment Analysis (CFA) to construct and analyze ligands composed of linking those fragments with predicted high affinity. This technique addresses the issue of effectively linking fragments together and provides a predictive mechanism to rank order prospective inhibitors for synthesis. The application of these techniques to the identification of novel inhibitors of human renin is described. Synthesis of a limited set of designed compounds provided potent, low MW analogs (IC50s<100nM) with good oral bioavailability (F>20-58%).
Asunto(s)
Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Renina/antagonistas & inhibidores , Animales , Disponibilidad Biológica , Diseño de Fármacos , Inhibidores Enzimáticos/síntesis química , Humanos , Ratas , Relación Estructura-Actividad , TermodinámicaRESUMEN
DAPK1 and ZIPK (also called DAPK3) are closely related serine/threonine protein kinases that regulate programmed cell death and phosphorylation of non-muscle and smooth muscle myosin. We have developed a fluorescence linked enzyme chemoproteomic strategy (FLECS) for the rapid identification of inhibitors for any element of the purinome and identified a selective pyrazolo[3,4-d]pyrimidinone (HS38) that inhibits DAPK1 and ZIPK in an ATP-competitive manner at nanomolar concentrations. In cellular studies, HS38 decreased RLC20 phosphorylation. In ex vivo studies, HS38 decreased contractile force generated in mouse aorta, rabbit ileum, and calyculin A stimulated arterial muscle by decreasing RLC20 and MYPT1 phosphorylation. The inhibitor also promoted relaxation in Ca(2+)-sensitized vessels. A close structural analogue (HS43) with 5-fold lower affinity for ZIPK produced no effect on cells or tissues. These findings are consistent with a mechanism of action wherein HS38 specifically targets ZIPK in smooth muscle. The discovery of HS38 provides a lead scaffold for the development of therapeutic agents for smooth muscle related disorders and a chemical means to probe the function of DAPK1 and ZIPK across species.
Asunto(s)
Proteínas Quinasas Asociadas a Muerte Celular/antagonistas & inhibidores , Proteínas Fluorescentes Verdes/metabolismo , Músculo Liso/efectos de los fármacos , Miocitos del Músculo Liso/efectos de los fármacos , Inhibidores de Proteínas Quinasas/farmacología , Pirazoles/farmacología , Pirimidinonas/farmacología , Proteínas Recombinantes de Fusión/metabolismo , Adenosina Trifosfato/metabolismo , Animales , Aorta/citología , Aorta/efectos de los fármacos , Aorta/enzimología , Unión Competitiva , Calcio/metabolismo , Proteínas Quinasas Asociadas a Muerte Celular/genética , Proteínas Quinasas Asociadas a Muerte Celular/metabolismo , Proteínas Fluorescentes Verdes/genética , Células HEK293 , Ensayos Analíticos de Alto Rendimiento , Humanos , Íleon/citología , Íleon/efectos de los fármacos , Íleon/enzimología , Ratones , Contracción Muscular/efectos de los fármacos , Músculo Liso/citología , Músculo Liso/enzimología , Miocitos del Músculo Liso/citología , Miocitos del Músculo Liso/enzimología , Quinasa de Cadena Ligera de Miosina/antagonistas & inhibidores , Quinasa de Cadena Ligera de Miosina/metabolismo , Fosfatasa de Miosina de Cadena Ligera , Fosforilación , Cultivo Primario de Células , Inhibidores de Proteínas Quinasas/química , Proteómica , Pirazoles/química , Pirimidinonas/química , Conejos , Proteínas Recombinantes de Fusión/genéticaRESUMEN
A novel class of Hsp90 inhibitors, structurally distinct from previously reported scaffolds, was developed from rational design and optimization of a compound library screen hit. These aminoquinazoline derivatives, represented by compound 15 (SNX-6833) or 1-(2-amino-4-methylquinazolin-7-yl)-3,6,6-trimethyl-6,7-dihydro-1H-indol-4(5H)-one, selectively bind to Hsp90 and inhibit its cellular activities at concentrations as low as single digit nanomolar.
Asunto(s)
Antineoplásicos/síntesis química , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Indoles/síntesis química , Quinazolinas/síntesis química , Antineoplásicos/farmacología , Línea Celular Tumoral , Proliferación Celular , Descubrimiento de Drogas , Ensayos de Selección de Medicamentos Antitumorales , Proteínas HSP90 de Choque Térmico/química , Humanos , Indoles/farmacología , Modelos Moleculares , Unión Proteica , Quinazolinas/farmacología , Bibliotecas de Moléculas Pequeñas , Relación Estructura-ActividadRESUMEN
Seeking compounds preferentially potent and selective for MMP-13, we reported in the preceding Letter on a series of hydroxamic acids with a flexible benzamide tail groups.(1a) Here, we replace the amide moiety with non-hydrolyzable heterocycles in an effort to improve half-life. We identify a hydroxamate tetrazole 4e that spares MMP-1 and -14, shows >400-fold selectivity versus MMP-8 and >600-fold selectivity versus MMP-2, and has a 4.8 h half-life in rats. X-ray data (1.9 Å) for tetrazole 4c is presented.
Asunto(s)
Amidas/síntesis química , Inhibidores Enzimáticos/síntesis química , Compuestos Heterocíclicos/síntesis química , Ácidos Hidroxámicos/síntesis química , Inhibidores de la Metaloproteinasa de la Matriz , Sulfonas/síntesis química , Amidas/química , Animales , Cristalografía por Rayos X , Inhibidores Enzimáticos/química , Compuestos Heterocíclicos/química , Ácidos Hidroxámicos/química , Metaloproteinasa 13 de la Matriz/química , Modelos Moleculares , Ratas , Relación Estructura-Actividad , Especificidad por Sustrato , Sulfonas/químicaRESUMEN
Continuing our interest in designing compounds preferentially potent and selective for MMP-13, we report on a series of hydroxamic acids with a flexible amide P1' substituents. We identify an amide which spares both MMP-1 and -14, and shows >500 fold selectivity for MMP-13 versus MMP-2 and -8.
Asunto(s)
Amidas/síntesis química , Ácidos Hidroxámicos/síntesis química , Inhibidores de la Metaloproteinasa de la Matriz , Inhibidores de Proteasas/síntesis química , Amidas/química , Humanos , Ácidos Hidroxámicos/química , Concentración 50 Inhibidora , Estructura Molecular , Inhibidores de Proteasas/química , Relación Estructura-Actividad , Especificidad por SustratoRESUMEN
α-Sulfone-α-piperidine and α-tetrahydropyranyl hydroxamates were explored that are potent inhibitors of MMP's-2, -9, and -13 that spare MMP-1, with oral efficacy in inhibiting tumor growth in mice and left-ventricular hypertrophy in rats and in the bovine cartilage degradation ex vivo explant system. α-Piperidine 19v (SC-78080/SD-2590) was selected for development toward the initial indication of cancer, while α-piperidine and α-tetrahydropyranyl hydroxamates 19w (SC-77964) and 9i (SC-77774), respectively, were identified as backup compounds.
Asunto(s)
Antiinflamatorios/síntesis química , Antineoplásicos/síntesis química , Fármacos Cardiovasculares/síntesis química , Ácidos Hidroxámicos/síntesis química , Inhibidores de la Metaloproteinasa de la Matriz , Piperidinas/síntesis química , Piranos/síntesis química , Sulfonas/síntesis química , Animales , Antiinflamatorios/química , Antiinflamatorios/farmacología , Antineoplásicos/química , Antineoplásicos/farmacología , Disponibilidad Biológica , Fármacos Cardiovasculares/química , Fármacos Cardiovasculares/farmacología , Cartílago Articular/efectos de los fármacos , Cartílago Articular/patología , Bovinos , Cristalografía por Rayos X , Humanos , Ácidos Hidroxámicos/química , Ácidos Hidroxámicos/farmacología , Hipertrofia Ventricular Izquierda/tratamiento farmacológico , Macaca fascicularis , Ratones , Ratones Desnudos , Modelos Moleculares , Estructura Molecular , Piperidinas/química , Piperidinas/farmacología , Piranos/química , Piranos/farmacología , Ratas , Relación Estructura-Actividad , Sulfonas/química , Sulfonas/farmacología , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
A chemoproteomics-based drug discovery strategy is presented that utilizes a highly parallel screening platform, encompassing more than 1000 targets, with a focused chemical library prior to target selection. This chemoproteomics-based process enables a data-driven selection of both the biological target and chemical hit after the screen is complete. The methodology has been exemplified for the purine binding proteome (proteins utilizing ATP, NAD, FAD). Screening of an 8000 member library yielded over 1500 unique protein-ligand interactions, which included novel hits for the oncology target Hsp90. The approach, which also provides broad target selectivity information, was used to drive the identification of a potent and orally active Hsp90 inhibitor, SNX-5422, which is currently in phase 1 clinical studies.
Asunto(s)
Evaluación Preclínica de Medicamentos/métodos , Proteínas HSP90 de Choque Térmico/metabolismo , Proteómica/métodos , Adenosina Trifosfato/metabolismo , Administración Oral , Animales , Unión Competitiva , Ensayos Clínicos Fase I como Asunto , Femenino , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Proteínas HSP90 de Choque Térmico/química , Humanos , Ratones , Modelos Moleculares , Conformación Molecular , Bibliotecas de Moléculas Pequeñas/administración & dosificación , Bibliotecas de Moléculas Pequeñas/química , Bibliotecas de Moléculas Pequeñas/metabolismo , Bibliotecas de Moléculas Pequeñas/farmacología , Especificidad por SustratoRESUMEN
A novel class of heat shock protein 90 (Hsp90) inhibitors was developed from an unbiased screen to identify protein targets for a diverse compound library. These indol-4-one and indazol-4-one derived 2-aminobenzamides showed strong binding affinity to Hsp90, and optimized analogues exhibited nanomolar antiproliferative activity across multiple cancer cell lines. Heat shock protein 70 (Hsp70) induction and specific client protein degradation in cells on treatment with the inhibitors supported Hsp90 inhibition as the mechanism of action. Computational chemistry and X-ray crystallographic analysis of selected member compounds clearly defined the protein-inhibitor interaction and assisted the design of analogues. 4-[6,6-Dimethyl-4-oxo-3-(trifluoromethyl)-4,5,6,7-tetrahydro-1H-indazol-1-yl]-2-[(trans-4-hydroxycyclohexyl)amino]benzamide (SNX-2112, 9) was identified as highly selective and potent (IC(50) Her2 = 11 nM, HT-29 = 3 nM); its prodrug amino-acetic acid 4-[2-carbamoyl-5-(6,6-dimethyl-4-oxo-3-trifluoromethyl-4,5,6,7-tetrahydro-indazol-1-yl)-phenylamino]-cyclohexyl ester methanesulfonate (SNX-5422, 10) was orally bioavailable and efficacious in a broad range of xenograft tumor models (e.g. 67% growth delay in a HT-29 model) and is now in multiple phase I clinical trials.
Asunto(s)
Antineoplásicos/administración & dosificación , Antineoplásicos/farmacología , Descubrimiento de Drogas , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , ortoaminobenzoatos/administración & dosificación , ortoaminobenzoatos/farmacología , Administración Oral , Animales , Antineoplásicos/química , Antineoplásicos/farmacocinética , Disponibilidad Biológica , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Ensayos Clínicos como Asunto , Femenino , Compuestos Heterocíclicos de 4 o más Anillos/química , Compuestos Heterocíclicos de 4 o más Anillos/farmacocinética , Compuestos Heterocíclicos de 4 o más Anillos/farmacología , Humanos , Concentración 50 Inhibidora , Ratones , Modelos Moleculares , Conformación Molecular , Profármacos/farmacocinética , Especificidad por Sustrato , ortoaminobenzoatos/química , ortoaminobenzoatos/farmacocinéticaRESUMEN
In the course of our Heat Shock 90 program, certain carbazole compounds were identified which had an off-target antiproliferative activity. To understand the off-target activity, we studied one analog with strong activity. We discovered that it had an effect on tubulin polymerization kinetics and was competitive with colchicine. Additional analogs were made, and a number of potent compounds were identified.
Asunto(s)
Antimitóticos/química , Carbazoles/química , Indoles/química , Antimitóticos/síntesis química , Antimitóticos/farmacología , Carbazoles/síntesis química , Carbazoles/farmacología , Línea Celular Tumoral , Colchicina/farmacología , Ensayos de Selección de Medicamentos Antitumorales , Proteínas HSP90 de Choque Térmico/metabolismo , Humanos , Indoles/síntesis química , Indoles/farmacología , Tubulina (Proteína)/metabolismoRESUMEN
OBJECTIVE: To evaluate the ability of SNX-7081, a novel small molecule inhibitor of Hsp90, to block components of inflammation, including cytokine production, protein kinase activity, and angiogenic signaling. A close analog was evaluated in preclinical in vivo models of rheumatoid arthritis (RA). METHODS: SNX-7081 binding to Hsp90 was characterized in Jurkat cells and RA synovial fibroblasts (RASFs). Inhibition of NF-kappaB nuclear translocation was evaluated in cellular systems, using lipopolysaccharide (LPS), tumor necrosis factor alpha, or interleukin-1beta stimulation. Suppression of cytokine production in THP-1 cells, human umbilical vein endothelial cells, and RASFs was studied. Disruption of MAPK signaling cascades by SNX-7081 following growth factor stimulation was assessed. SNX-7081 was tested in 2 relevant angiogenesis assays: platelet-derived growth factor activation of fibroblasts and LPS-induced nitric oxide (NO) release in J774 macrophages. A close analog, SNX-4414, was evaluated in rat collagen-induced arthritis and adjuvant-induced arthritis, following oral treatment. RESULTS: SNX-7081 showed strong binding affinity to Hsp90 and expected induction of Hsp70. NF-kappaB nuclear translocation was blocked by SNX-7081 at nanomolar concentrations, and cytokine production was potently inhibited. Growth factor activation of ERK and JNK signaling was significantly reduced by SNX-7081. NO production was also sharply inhibited. In animal models, SNX-4414 fully inhibited paw swelling and improved body weight. Scores for inflammation, pannus formation, cartilage damage, and bone resorption returned to normal. CONCLUSION: The present results demonstrate that a small molecule Hsp90 inhibitor can impact inflammatory disease processes. The strong in vivo efficacy observed with SNX-4414 provides preclinical validation for consideration of Hsp90 inhibitors in the treatment of RA.
Asunto(s)
Antiinflamatorios/farmacología , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/metabolismo , Benzamidas/farmacología , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Transducción de Señal/efectos de los fármacos , Administración Oral , Animales , Antiinflamatorios/farmacocinética , Artritis Reumatoide/inmunología , Benzamidas/farmacocinética , Citocinas/metabolismo , Modelos Animales de Enfermedad , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/inmunología , Femenino , Fibroblastos/citología , Proteínas del Choque Térmico HSP72/metabolismo , Humanos , Células Jurkat , Macrófagos/citología , Masculino , Ratones , Proteínas Quinasas Activadas por Mitógenos/metabolismo , FN-kappa B/metabolismo , Células 3T3 NIH , Neovascularización Fisiológica/fisiología , Óxido Nítrico/metabolismo , Ratas , Ratas Endogámicas Lew , Ratas Sprague-Dawley , Transducción de Señal/inmunología , Membrana Sinovial/citología , omega-ConotoxinasRESUMEN
Hsp90 maintains the conformational stability of multiple proteins implicated in oncogenesis and has emerged as a target for chemotherapy. We report here the discovery of a novel small molecule scaffold that inhibits Hsp90. X-ray data show that the scaffold binds competitively at the ATP site on Hsp90. Cellular proliferation and client assays demonstrate that members of the series are able to inhibit Hsp90 at nanomolar concentrations.
Asunto(s)
Antineoplásicos/farmacología , Carbazoles/farmacología , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Adenosina Trifosfato/química , Antineoplásicos/síntesis química , Antineoplásicos/química , Unión Competitiva , Carbazoles/síntesis química , Carbazoles/química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Cristalografía por Rayos X , Ensayos de Selección de Medicamentos Antitumorales , Proteínas HSP90 de Choque Térmico/química , Humanos , Modelos Moleculares , Estructura Molecular , Peso Molecular , Bibliotecas de Moléculas Pequeñas , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
alpha-Piperidine-beta-sulfone hydroxamate derivatives were explored that are potent for matrix metalloproteinases (MMP)-2, -9, and -13 and are sparing of MMP-1. The investigation of the beta-sulfones subsequently led to the discovery of hitherto unknown alpha-sulfone hydroxamates that are superior to the corresponding beta-sulfones in potency for target MMPs, selectivity vs MMP-1, and exposure when dosed orally. alpha-Piperidine-alpha-sulfone hydroxamate 35f (SC-276) was advanced through antitumor and antiangiogenesis assays and was selected for development. Compound 35f demonstrates excellent antitumor activity vs MX-1 breast tumor in mice when dosed orally as monotherapy or in combination with paclitaxel.