RESUMO
A series of novel C(1) alkylphosphinic acid analogues of the prostaglandin-F family have been evaluated at the eight human prostaglandin receptors for potential use in the treatment of osteoporosis. Using molecular modeling as a tool for structure-based drug design, we have discovered that the phosphinic acid moiety (P(O)(OH)R) behaves as an isostere for the C(1) carboxylic acid in the human prostaglandin FP binding assay in vitro and possesses enhanced hFP receptor selectivity when compared to the parent carboxylic acid. When evaluated in vivo, the methyl phosphinic acid analogue (4b) produced a bone anabolic response in rats, returning bone mineral volume (BMV) [corrected], to intact levels in the distal femur in the ovariectomized rat (OVX) model. These results suggest that prostaglandins of this class may be useful agents in the treatment of diseases associated with bone loss.
Assuntos
Osso e Ossos/efeitos dos fármacos , Dinoprosta/síntese química , Ácidos Fosfínicos/síntese química , Prostaglandinas F Sintéticas/síntese química , Absorciometria de Fóton , Sequência de Aminoácidos , Animais , Ligação Competitiva , Densidade Óssea/efeitos dos fármacos , Osso e Ossos/diagnóstico por imagem , Osso e Ossos/metabolismo , Células COS , Dinoprosta/análogos & derivados , Dinoprosta/química , Dinoprosta/metabolismo , Dinoprosta/farmacologia , Feminino , Humanos , Modelos Moleculares , Dados de Sequência Molecular , Osteoporose/tratamento farmacológico , Ovariectomia , Ácidos Fosfínicos/química , Ácidos Fosfínicos/metabolismo , Ácidos Fosfínicos/farmacologia , Prostaglandinas F Sintéticas/química , Prostaglandinas F Sintéticas/metabolismo , Prostaglandinas F Sintéticas/farmacologia , Ensaio Radioligante , Ratos , Ratos Sprague-Dawley , Receptores de Prostaglandina/metabolismo , Relação Estrutura-Atividade , Tomografia Computadorizada por Raios X , TransfecçãoAssuntos
Compostos de Bifenilo/síntese química , Ácidos Carboxílicos/síntese química , Inibidores de Metaloproteinases de Matriz , Inibidores de Proteases/síntese química , Compostos de Bifenilo/química , Proteínas Sanguíneas/química , Ácidos Carboxílicos/química , Inibidores de Proteases/química , Solubilidade , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
Potent and selective inhibition of matrix metalloproteinases was demonstrated for a series of sulfonamide-based hydroxamic acids. The design of the heterocyclic sulfonamides incorporates a six- or seven-member central ring with a P2' substituent that can be modified. Binding interactions of this substituent at the S2' site are believed to contribute to high inhibitory potency against stromelysin, collagenase-3 and gelatinases A and B, and to provide selectivity against collagenase-1 and matrilysin. An X-ray structure of a stromelysin inhibitor complex was obtained to provide insights into the SAR and selectivity trends observed for the series.
Assuntos
Compostos Heterocíclicos com 1 Anel/farmacologia , Inibidores de Metaloproteinases de Matriz , Sulfonamidas/farmacologia , Colagenases , Cristalografia por Raios X , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/farmacologia , Compostos Heterocíclicos com 1 Anel/síntese química , Concentração Inibidora 50 , Substâncias Macromoleculares , Metaloproteinase 13 da Matriz , Sensibilidade e Especificidade , Relação Estrutura-Atividade , Sulfonamidas/síntese químicaRESUMO
A series of carboxylic acids were prepared from a propargylglycine scaffold and tested for efficacy as matrix metalloproteinase (MMP) inhibitors. Detailed SAR for the series is reported for four enzymes within the MMP family. The inhibitors were typically potent against collagenase-3 (MMP-13) and gelatinase A (MMP-2), while they spared collagenase-1 (MMP-1) and only moderately inhibited stromelysin (MMP-3). Compound 40 represents a typical inhibition profile of a compound with reasonable potency. Introduction of polar groups was required in order to generate inhibitors with acceptable water solubility, and this often resulted in a loss of potency as in compound 63. High serum protein binding proved to be a difficult hurdle with many compounds such as 48 showing >99% binding. Some compounds such as 64 displayed approximately 90% binding, but no reliable method was discovered for designing molecules with low protein binding. Finally, selected data regarding the pharmacokinetic behavior of these compounds is presented.
Assuntos
Alcinos/síntese química , Ácidos Carboxílicos/síntese química , Glicina/análogos & derivados , Glicina/síntese química , Metaloendopeptidases/antagonistas & inibidores , Inibidores de Proteases/síntese química , Alcinos/química , Ácidos Carboxílicos/química , Glicina/química , Humanos , Metaloproteinase 3 da Matriz/química , Inibidores de Metaloproteinases de Matriz , Metaloendopeptidases/química , Modelos Moleculares , Inibidores de Proteases/química , Relação Estrutura-AtividadeRESUMO
The in vitro evaluation of a new class of potential bone anabolic agents for the treatment of osteoporosis is described. These compounds are potent and selective ligands for the human prostaglandin F receptor (hFP receptor). The compounds lack the olefin unsaturation required for potency in the natural ligand PGF(2)(alpha) yet retain binding affinity for the hFP receptor in the nanomolar to micromolar range. Removal of the alkenes also results in a better selectivity ratio for the hFP receptor over the other prostaglandin receptors tested. A rationale for the selectivity differences of various analogues, based on ligand docking experiments to a putative hFP receptor model, is also described.
Assuntos
Prostaglandinas F/síntese química , Receptores de Prostaglandina/metabolismo , Animais , Ligação Competitiva , Células COS , Desenho de Fármacos , Humanos , Ligantes , Modelos Moleculares , Osteoporose/tratamento farmacológico , Prostaglandinas F/química , Prostaglandinas F/metabolismo , Prostaglandinas F/farmacologia , Ensaio Radioligante , Ratos , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
A new generation of cyclic matrix metalloproteinase (MMP) inhibitors derived from dl-piperazinecarboxylic acid has been described. The design involves: incorporation of hydroxamic acid as the bidentate chelating agent for catalytic Zn(2+), placement of a sulfonamide group at the 1N-position of the piperazine ring to fill the S1' pocket of the enzyme, and finally attachment of diverse functional groups at the 4N-position to optimize potency and peroral absorption. A unique combination of all three elements produced inhibitor 20 with high affinity for MMPs 1, 3, 9, and 13 (24, 18, 1.9, and 1.3 nM, respectively). X-ray crystallography data obtained for MMP-3 cocrystallized with 20 gave detailed information on key binding interactions defining an overall scaffold geometry for piperazine-based MMP inhibitors.
Assuntos
Metaloendopeptidases/antagonistas & inibidores , Piperazinas/síntese química , Inibidores de Proteases/síntese química , Sulfonamidas/síntese química , Animais , Cartilagem/citologia , Cartilagem/efeitos dos fármacos , Bovinos , Células Cultivadas , Cristalografia por Raios X , Desenho de Fármacos , Modelos Moleculares , Piperazinas/química , Piperazinas/farmacologia , Inibidores de Proteases/química , Inibidores de Proteases/farmacologia , Proteínas Recombinantes/antagonistas & inibidores , Estereoisomerismo , Relação Estrutura-Atividade , Sulfonamidas/química , Sulfonamidas/farmacologiaRESUMO
A series of hydroxamates was prepared from an aminoproline scaffold and tested for efficacy as matrix metalloproteinase (MMP) inhibitors. Detailed SAR for the series is reported for five enzymes within the MMP family, and a number of inhibitors, such as compound 47, display broad-spectrum activity with sub-nanomolar potency for some enzymes. Modifications of the P1' portion of the molecule played a key role in affecting both potency and selectivity within the MMP family. Longer-chain aliphatic substituents in this region of the molecule tended to increase potency for MMP-3 and decrease potency for MMP-1, as exemplified by compounds 48-50, while aromatic substituents, as in compound 52, generated broad-spectrum inhibition. The data is rationalized based upon X-ray crystal data which is also presented. While the in vitro peroral absorption seemed to be less predictable, it tended to decrease with longer and more hydrophilic substituents. Finally, a rat model of osteoarthritis was used to evaluate the efficacy of these compounds, and a direct link was established between their pharmacokinetics and their in vivo efficacy.
Assuntos
Ácidos Hidroxâmicos/síntese química , Metaloendopeptidases/antagonistas & inibidores , Prolina/análogos & derivados , Prolina/síntese química , Inibidores de Proteases/síntese química , Animais , Cartilagem Articular/patologia , Cristalografia por Raios X , Humanos , Ácidos Hidroxâmicos/química , Ácidos Hidroxâmicos/farmacologia , Iodoacetatos , Masculino , Metaloproteinase 3 da Matriz/química , Modelos Moleculares , Osteoartrite do Joelho/induzido quimicamente , Osteoartrite do Joelho/tratamento farmacológico , Osteoartrite do Joelho/patologia , Prolina/química , Prolina/farmacologia , Inibidores de Proteases/química , Inibidores de Proteases/farmacologia , Ratos , Ratos Sprague-Dawley , Relação Estrutura-AtividadeRESUMO
The synthesis and enzyme inhibition data for a series of thiazine- and thiazepine-based matrix metalloproteinase (MMP) inhibitors are described. The thiazine- and thiazepine-based inhibitors were discovered by optimization of hetererocyclic sulfonamide-based inhibitors. The most potent series of inhibitors was obtained by modification of the amino acid D-penicillamine. This amino acid provides a gem-dimethyl group on the thiazine or thiazepine ring which has a dramatic effect on the in vitro potency of this series. In particular, the sulfide 4a and the sulfone 5a were potent, broad-spectrum inhibitors of the MMPs with IC(50)'s against MMP-1 of 0.8 and 1.9 nM, respectively. The binding mode of this novel thiazepine-based series of MMP inhibitors was established based on X-ray crystallography of the complex of stromelysin and 4a.
Assuntos
Óxidos S-Cíclicos/síntese química , Inibidores Enzimáticos/síntese química , Metaloendopeptidases/antagonistas & inibidores , Tiazepinas/síntese química , Tiazinas/síntese química , Cristalografia por Raios X , Óxidos S-Cíclicos/química , Desenho de Fármacos , Inibidores Enzimáticos/química , Modelos Moleculares , Relação Estrutura-Atividade , Tiazepinas/química , Tiazinas/químicaRESUMO
A new series of hydroxamic acid-based matrix metalloproteinase (MMP) inhibitors containing a unique phosphinamide motif derived from D-amino acid was designed, synthesized, and tested for enzyme inhibition. Compounds with an R configuration at phosphorus were found to be potent MMP inhibitors while molecules with the S configuration were almost inactive. Structure-activity relationship studies of the series led to the discovery of the potent inhibitor 16 with IC50 = 20.5 nM and 24.4 nM against fibroblast collagenase (MMP-1) and stromelysin (MMP-3), respectively. The binding mode of this novel phosphinamide-based series of MMP inhibitors was established based on X-ray crystallography of the complex of stromelysin and 16.
Assuntos
Ácidos Hidroxâmicos/síntese química , Metaloendopeptidases/antagonistas & inibidores , Compostos Organofosforados/síntese química , Inibidores de Proteases/síntese química , Sítios de Ligação , Cristalografia por Raios X , Desenho de Fármacos , Humanos , Ácidos Hidroxâmicos/química , Ácidos Hidroxâmicos/metabolismo , Ácidos Hidroxâmicos/farmacologia , Metaloproteinase 1 da Matriz , Metaloproteinase 13 da Matriz , Metaloproteinase 3 da Matriz/metabolismo , Metaloproteinase 7 da Matriz , Metaloproteinase 9 da Matriz , Inibidores de Metaloproteinases de Matriz , Modelos Moleculares , Compostos Organofosforados/química , Compostos Organofosforados/metabolismo , Compostos Organofosforados/farmacologia , Inibidores de Proteases/química , Inibidores de Proteases/metabolismo , Inibidores de Proteases/farmacologia , Proteínas Recombinantes/antagonistas & inibidores , Estereoisomerismo , Relação Estrutura-AtividadeAssuntos
Metaloendopeptidases/antagonistas & inibidores , Piperazinas/síntese química , Cristalografia por Raios X , Metaloproteinase 1 da Matriz , Metaloproteinase 3 da Matriz/metabolismo , Metaloproteinase 7 da Matriz , Metaloproteinase 9 da Matriz , Inibidores de Metaloproteinases de Matriz , Metaloendopeptidases/metabolismo , Modelos Moleculares , Conformação Molecular , Piperazinas/química , Piperazinas/metabolismo , Piperazinas/farmacologia , Inibidores de Proteases/síntese química , Inibidores de Proteases/química , Inibidores de Proteases/metabolismo , Inibidores de Proteases/farmacologia , Ligação Proteica , Relação Estrutura-AtividadeRESUMO
A novel series of conformationally constrained matrix metalloprotease inhibitors was identified. The potencies observed for these inhibitors were highly dependent upon the substitution pattern on the caprolactam ring as well as the succinate moiety.
Assuntos
Caprolactama/análogos & derivados , Caprolactama/síntese química , Metaloendopeptidases/antagonistas & inibidores , Inibidores de Proteases/química , Inibidores de Proteases/síntese química , Caprolactama/química , Caprolactama/farmacologia , Colagenases/química , Desenho de Fármacos , Indicadores e Reagentes , Cinética , Metaloproteinase 1 da Matriz , Metaloproteinase 3 da Matriz/química , Inibidores de Metaloproteinases de Matriz , Modelos Moleculares , Conformação Molecular , Estrutura Molecular , Inibidores de Proteases/farmacologia , Conformação Proteica , Relação Estrutura-Atividade , Succinatos/síntese química , Succinatos/química , Succinatos/farmacologiaRESUMO
We have adapted for glucose determination a new approach to kinetic analyses [Anal. Chem. 50, 1611 (1978)]; it is 50-fold less dependent upon some experimental variables than is a more conventional rate method. Modification of a commercially available hexokinase/glucose-6-phosphate dehydrogenase reagent system for glucose provides that the rate of production of NADH be first-order in total glucose concentration within about 30 s after sample and reagent are mixed. In the kinetic method, absorbance vs. time data recorded after 30 s and a multiple-linear-regression program are used to compute the absorbance change that would occur if the reaction were monitored to completion. Results demonstrate a linear relationship between glucose concentration and computed absorbance change. Application of the method to 51 human sera without rigorous control of either temperature or reagent composition yielded a regression equation of y = 1.01x -0.3 when kinetic results (y) were compared with equilibrium results (x) for the same samples analyzed in a hospital laboratory.
Assuntos
Glicemia/análise , Glucosefosfato Desidrogenase/metabolismo , Hexoquinase/metabolismo , Humanos , Cinética , Análise de Regressão , Espectrofotometria Ultravioleta/métodos , TemperaturaRESUMO
This paper describes the evaluation of a system for computer-controlled discrete sampling and stopped-flow mixing for equilibrium and kinetic determinations of several sorts of analytes in human serum. The instrumental system features a wash-out sampling system that permits rapid change-over from one sample and (or) reagent type to another, and a mixing-measurement system that can provide reliable data as soon as 10 ms after reagent and sample are mixed. Examples discussed include equilibrium procedures for glucose and cholesterol, slow kinetic procedures for glucose and lactate dehydrogenase, and a fast kinetic method for thiocyanate. The regression equation for all stopped-flow results (n = 114) vs. results by conventional methods is y = (103 +/- 0.01)x - (0.016 +/- 0.019) for numerical values of y between 0.3 and 3.0. The correlation coefficient for these data was 0.991. These results demonstrate that the stopped-flow method is a viable analytical approach for equilibrium, slow kinetic, and fast kinetic determinations that require measurement times shorter than 0.1 s.