RESUMEN
TAK-599 (known as ceftaroline fosamil) is a novel N-phosphono type prodrug of a cephalosporin compound, T-91825, that exhibits strong activity against methicillin resistant Staphylococcus aureus (MRSA). The stability and stabilization of TAK-599 were investigated by kinetic analysis focused on crystallinity and moisture content. Initially it was planned to develop TAK-599 as an injectable formulation using the amorphous solid powder prepared by lyophilization. However, amorphous of TAK-599 free form was found to be chemically unstable even when stored at 8 degrees C, and thus development was focused on the crystalline material. After exhaustive screening of crystallization condition, the monoacetic acid solvate was found to yield TAK-599 in a crystalline form. Physicochemical properties were studied to identify the key factors affecting the stabilization of TAK-599 in order to improve long-term stability, and the results indicated that the crystallinity of TAK-599 correlated with stability. Furthermore, moisture content was also identified in our studies as an important factor in stabilizing TAK-599. TAK-599 containing about 3% moisture was found to be the most stable form. It was concluded that both sufficient crystallinity and strict moisture control of TAK-599 were essential to maintain long-term stability at 25 degrees C.
Asunto(s)
Antibacterianos/química , Antibacterianos/farmacología , Cefalosporinas/química , Cefalosporinas/farmacología , Resistencia a la Meticilina/efectos de los fármacos , Profármacos/química , Profármacos/farmacología , Staphylococcus aureus/efectos de los fármacos , Fenómenos Químicos , Química Física , Cromatografía Líquida de Alta Presión , Estabilidad de Medicamentos , Liofilización , Solubilidad , Espectrofotometría Ultravioleta , Agua/análisis , Difracción de Rayos X , CeftarolinaRESUMEN
Replacement of the 5-oxopyrrolidin-3-yl fragment in the previously reported lead structure with a 1-acetylpiperidin-4-yl group led to the discovery of a novel series of potent CCR5 antagonists. Introduction of small hydrophobic substituents on the central phenyl ring increased the binding affinity, providing low to sub-nanomolar CCR5 antagonists. The selected compound 11f showed excellent antiviral activity against CCR5-using HIV-1 replication in human peripheral blood mononuclear cells (EC50=0.59 nM) and an acceptable pharmacokinetic profile in dogs.
Asunto(s)
Fármacos Anti-VIH/síntesis química , Antagonistas de los Receptores CCR5 , Piperidinas/síntesis química , Animales , Fármacos Anti-VIH/química , Fármacos Anti-VIH/farmacocinética , Fármacos Anti-VIH/farmacología , Células CHO , Células COS , Chlorocebus aethiops , Cricetinae , Cricetulus , Perros , VIH-1/efectos de los fármacos , Humanos , Masculino , Modelos Químicos , Modelos Moleculares , Estructura Molecular , Piperidinas/química , Piperidinas/farmacocinética , Piperidinas/farmacología , Ratas , Receptores CCR5/química , Relación Estructura-ActividadRESUMEN
TAK-599 is a water-soluble prodrug of a cephalosporin compound, T-91825. In vitro and in vivo antibacterial activities of T-91825 and TAK-599, respectively, were examined. T-91825 was active against both gram-positive and gram-negative bacteria, unlike vancomycin and linezolid, which are inactive against gram-negative bacteria. The 90% minimum inhibitory concentration of T-91825 against clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) was 2 micro g/ml. This activity was comparable to those of vancomycin, linezolid, teicoplanin, and arbekacin. T-91825 was similarly active against vancomycin-intermediate S. aureus. In a time-kill study, T-91825 showed more rapid and distinct decrease of viable cells of two MRSA strains than did vancomycin and linezolid in vitro. The effect of TAK-599 against systemic infection caused by clinical isolates of MRSA in mice was comparable or superior to that of vancomycin, linezolid, teicoplanin, and arbekacin. In addition, TAK-599 at a dose of 20 mg/kg significantly decreased bacterial counts in lungs of mice in an experimental pneumonia model caused by MRSA in which vancomycin and linezolid were totally ineffective at the same dose. These results suggest the usefulness of TAK-599 in the treatment of MRSA infections in humans.
Asunto(s)
Butiratos/farmacología , Cefalosporinas/farmacología , Resistencia a la Meticilina , Oxazoles/farmacología , Staphylococcus/efectos de los fármacos , Animales , Butiratos/uso terapéutico , Cefalosporinas/uso terapéutico , Modelos Animales de Enfermedad , Enterococcus/clasificación , Enterococcus/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos CBA , Ratones Endogámicos ICR , Pruebas de Sensibilidad Microbiana , Oxazoles/uso terapéutico , Neumonía Bacteriana/tratamiento farmacológico , Neumonía Bacteriana/microbiología , Infecciones Estafilocócicas/tratamiento farmacológico , Infecciones Estafilocócicas/microbiología , Staphylococcus/clasificación , CeftarolinaRESUMEN
We have previously reported the novel lead compound 1a as a CCR5 antagonist for treatment of HIV-1 infection. SAR studies on incorporating various acyl groups as a replacement for the 5-oxopyrrolidine-3-carbonyl group of the lead structure resulted in the discovery of N-[3-(4-benzylpiperidin-1-yl)propyl]-N,N'-diphenylurea (4a) with significantly improved CCR5 binding affinity. Substitutions (4-Cl, 4e,f; 4-Me, 4i) on the N'-phenyl ring further increased the binding affinity. Introduction of polar substituents on the phenyl ring of the 4-benzylpiperidine moiety enhanced the inhibitory activity of the HIV-1 envelope-mediated membrane fusion (4v,w), suggesting that polar substituents at this position can interfere effectively with HIV-1 cell entry.
Asunto(s)
Fármacos Anti-VIH/síntesis química , Fármacos Anti-VIH/farmacología , Antagonistas de los Receptores CCR5 , Carbanilidas/síntesis química , Carbanilidas/farmacología , Animales , Fármacos Anti-VIH/química , Células CHO , Carbanilidas/química , Cricetinae , VIH-1/efectos de los fármacos , VIH-1/fisiología , Espectroscopía de Resonancia Magnética , Fusión de Membrana/efectos de los fármacosRESUMEN
A novel lead compound, N-(3-[4-(4-fluorobenzoyl)piperidin-1-yl]propyl)-1-methyl-5-oxo-N-phenylpyrrolidine-3-carboxamide (1), was identified as a CCR5 antagonist by high-throughput screening using [(125)I]RANTES and CCR5-expressing CHO cells. The IC(50) value of 1 was 1.9 microM. In an effort to improve the binding affinity of 1, a series of 5-oxopyrrolidine-3-carboxamides was synthesized. Introduction of 3,4-dichloro substituents to the central phenyl ring (10i, IC(50)=0.057 microM; 11b, IC(50)=0.050 microM) or replacing the 1-methyl group of the 5-oxopyrrolidine moiety with a 1-benzyl group (12e, IC(50)=0.038 microM) was found to be effective for improving CCR5 affinity. Compound 10i, 11b, and 12e also inhibited CCR5-using HIV-1 envelope-mediated membrane fusion with IC(50) values of 0.44, 0.19, and 0.49 microM, respectively.
Asunto(s)
Amidas/química , Amidas/farmacología , Fármacos Anti-VIH/síntesis química , Fármacos Anti-VIH/farmacología , Antagonistas de los Receptores CCR5 , Pirrolidinas/química , Pirrolidinas/farmacología , Animales , Células CHO , Células COS , Quimiocina CCL5/antagonistas & inhibidores , Chlorocebus aethiops , Cricetinae , Evaluación Preclínica de Medicamentos , Femenino , VIH-1/efectos de los fármacos , Humanos , Indicadores y Reactivos , Espectroscopía de Resonancia Magnética , Piperidinas/química , Piperidinas/farmacología , Relación Estructura-ActividadRESUMEN
Crystalline 1 (TAK-599) is a novel N-phosphono prodrug of anti-methicillin-resistant Staphylococcus aureus (MRSA) cephalosporin 2a (T-91825) that has high affinity for penicillin-binding protein (PBP) 2' (IC(50); 0.90 microg/mL) and shows potent in vitro anti-MRSA activity (MIC against MRSA N133; 1.56 microg/mL), comparable to that of vancomycin (1.56 microg/mL). Although 2a had insufficient water solubility (2.3 mg/mL) for parenteral administration, 1 showed excellent water solubility (>100 mg/mL, pH 7) as well as good chemical stability in the solid state and solution. In pharmacokinetic studies, when 1 was administered intravenously to rats and monkeys, it was rapidly converted into 2a in the blood. These results show that 1 (TAK-599) is a highly promising parenteral cephalosporin targeted for MRSA infection.