RESUMO
A novel class of timolol derivatives with nitric oxide (NO)-donating moieties achieved chemical stability yet under physiologically relevant conditions released timolol and NO. Hindered esters A were designed and synthesized, whose 'triggered' release relied on enzymatic hydrolysis of the nitrate ester in A to B, that in turn cyclized to liberate timolol.
Assuntos
Óxido Nítrico/metabolismo , Timolol/química , Antagonistas Adrenérgicos beta/metabolismo , Estabilidade de Medicamentos , Ésteres/química , Ésteres/metabolismo , Guanilato Ciclase/antagonistas & inibidores , Guanilato Ciclase/metabolismoRESUMO
The structure-based design, chemical synthesis, and biological evaluation of various 2-pyridone-containing human rhinovirus (HRV) 3C protease (3CP) inhibitors are described. These compounds are comprised of a peptidomimetic binding determinant and a Michael acceptor moiety, which forms an irreversible covalent adduct with the active site cysteine residue of the 3C enzyme. The 2-pyridone-containing inhibitors typically display improved 3CP inhibition properties relative to related peptide-derived molecules along with more favorable antiviral properties. The cocrystal structure of one pyridone-derived 3CP inhibitor complexed with HRV-2 3CP is also described along with certain ab initio conformation analyses. Optimization of the 2-pyridone-containing compounds is shown to provide several highly active 3CP inhibitors (k(obs)/[I] > 500,00 M(-1) s(-1)) that function as potent antirhinoviral agents (EC(50) = <0.05 microM) against multiple virus serotypes in cell culture. One 2-pyridone-containing 3CP inhibitor is shown to be bioavailable in the dog after oral dosing (F = 48%).
Assuntos
Antivirais/síntese química , Peptídeos/química , Inibidores de Proteases/síntese química , Piridonas/síntese química , Rhinovirus/enzimologia , Proteínas Virais/antagonistas & inibidores , Proteases Virais 3C , Administração Oral , Animais , Antivirais/química , Antivirais/farmacologia , Disponibilidade Biológica , Cristalografia por Raios X , Cisteína Endopeptidases , Cães , Estabilidade de Medicamentos , Humanos , Técnicas In Vitro , Ligantes , Microssomos Hepáticos/metabolismo , Modelos Moleculares , Mimetismo Molecular , Inibidores de Proteases/química , Inibidores de Proteases/farmacologia , Ligação Proteica , Piridonas/química , Piridonas/farmacologia , Relação Estrutura-AtividadeRESUMO
The optimization of the pharmacokinetic performance of various 2-pyridone-containing human rhinovirus (HRV) 3C protease (3CP) inhibitors following oral administration to either beagle dogs or CM-monkeys is described. The molecules described in this work are composed of a 2-pyridone-containing peptidomimetic binding determinant and an alpha,beta-unsaturated ester Michael acceptor moiety which forms an irreversible covalent adduct with the active site cysteine residue of the 3C enzyme. Modification of the ester contained within these compounds is detailed along with alteration of the P(2) substituent present in the peptidomimetic portion of the inhibitors. The pharmacokinetics of several inhibitors in both dogs and monkeys are described (7 h plasma concentrations after oral administration) along with their human plasma stabilities, stabilities in incubations with human, dog, and monkey microsomes and hepatocytes, Caco-2 permeabilities, and aqueous solubilities. Compounds containing an alpha,beta-unsaturated ethyl ester fragment and either an ethyl or propargyl P(2) moiety displayed the most promising combination of 3C enzyme inhibition (k(obs)/[I] 170 000-223 000 M(-1) s(-1)), antiviral activity (EC(50) = 0.047-0.058 microM, mean vs seven HRV serotypes), and pharmacokinetics following oral administration (7 h dog plasma levels = 0.248-0.682 microM; 7 h CM-monkey plasma levels = 0.057-0.896 microM).
Assuntos
Antivirais/síntese química , Antivirais/farmacologia , Cisteína Endopeptidases/metabolismo , Inibidores de Proteases/síntese química , Inibidores de Proteases/farmacologia , Piridonas/síntese química , Piridonas/farmacologia , Rhinovirus/enzimologia , Proteínas Virais/metabolismo , Proteases Virais 3C , Animais , Antivirais/farmacocinética , Disponibilidade Biológica , Proteínas Sanguíneas/metabolismo , Células CACO-2 , Cães , Desenho de Fármacos , Meia-Vida , Hepatócitos/metabolismo , Humanos , Técnicas In Vitro , Indicadores e Reagentes , Macaca fascicularis , Espectroscopia de Ressonância Magnética , Masculino , Microssomos Hepáticos/metabolismo , Inibidores de Proteases/farmacocinética , Ligação Proteica , Rhinovirus/efeitos dos fármacos , Solubilidade , Relação Estrutura-AtividadeRESUMO
PURPOSE: Nitric oxide (NO) is involved in a variety of physiological processes including ocular aqueous humor dynamics by targeting mechanisms that are complementary to those of prostaglandins. Here, we have characterized a newly synthesized compound, NCX 125, comprising latanoprost acid and NO-donating moieties. METHODS: NCX 125 was synthesized and tested in vitro for its ability to release functionally active NO and then compared with core latanoprost for its intraocular pressure (IOP)-lowering effects in rabbit, dog, and nonhuman primate models of glaucoma. RESULTS: NCX 125 elicited cGMP formation (EC(50) = 3.8 + or - 1.0 microM) in PC12 cells and exerted NO-dependent iNOS inhibition (IC(50) = 55 + or - 11 microM) in RAW 264.7 macrophages. NCX 125 lowered IOP to a greater extent compared with equimolar latanoprost in: (a) rabbit model of transient ocular hypertension (0.030% latanoprost, not effective; 0.039% NCX 125, Delta(max) = -10.6 + or - 2.3 mm Hg), (b) ocular hypertensive glaucomatous dogs (0.030% latanoprost, Delta(max)= -6.7 + or - 1.2 mm Hg; 0.039% NCX 125, Delta(max) = -9.1 + or - 3.1 mm Hg), and (c) laser-induced ocular hypertensive non-human primates (0.10% latanoprost, Delta(max) = -11.9 + or - 3.7 mm Hg, 0.13% NCX 125, Delta(max) = -16.7 + or - 2.2 mm Hg). In pharmacokinetic studies, NCX 125 and latanoprost resulted in similar latanoprost-free acid exposure in anterior segment ocular tissues. CONCLUSIONS: NCX 125, a compound targeting 2 different mechanisms, is endowed with potent ocular hypotensive effects. This may lead to potential new perspectives in the treatment of patients at risk of glaucoma.