RESUMO
Lipids are used as cellular building blocks and condensed energy stores and also act as signaling molecules. The glycerolipid/ fatty acid cycle, encompassing lipolysis and lipogenesis, generates many lipid signals. Reliable procedures are not available for measuring activities of several lipolytic enzymes for the purposes of drug screening, and this resulted in questionable selectivity of various known lipase inhibitors. We now describe simple assays for lipolytic enzymes, including adipose triglyceride lipase (ATGL), hormone sensitive lipase (HSL), sn-1-diacylglycerol lipase (DAGL), monoacylglycerol lipase, α/ß-hydrolase domain 6, and carboxylesterase 1 (CES1) using recombinant human and mouse enzymes either in cell extracts or using purified enzymes. We observed that many of the reported inhibitors lack specificity. Thus, Cay10499 (HSL inhibitor) and RHC20867 (DAGL inhibitor) also inhibit other lipases. Marked differences in the inhibitor sensitivities of human ATGL and HSL compared with the corresponding mouse enzymes was noticed. Thus, ATGListatin inhibited mouse ATGL but not human ATGL, and the HSL inhibitors WWL11 and Compound 13f were effective against mouse enzyme but much less potent against human enzyme. Many of these lipase inhibitors also inhibited human CES1. Results describe reliable assays for measuring lipase activities that are amenable for drug screening and also caution about the specificity of the many earlier described lipase inhibitors.
Assuntos
Descoberta de Drogas/métodos , Inibidores Enzimáticos/farmacologia , Lipólise/efeitos dos fármacos , Tecido Adiposo/metabolismo , Animais , Hidrolases de Éster Carboxílico/antagonistas & inibidores , Hidrolases de Éster Carboxílico/metabolismo , Ácidos Graxos/metabolismo , Humanos , Lipase/antagonistas & inibidores , Lipase/metabolismo , Lipogênese/fisiologia , Lipólise/fisiologia , Lipase Lipoproteica/antagonistas & inibidores , Lipase Lipoproteica/metabolismo , Camundongos , Monoacilglicerol Lipases/metabolismo , Esterol Esterase/antagonistas & inibidores , Esterol Esterase/metabolismo , Triglicerídeos/metabolismoRESUMO
ATR is a key kinase in the DNA-damage response (DDR) that is synthetic lethal with several other DDR proteins, making it an attractive target for the treatment of genetically selected solid tumors. Herein we describe the discovery of a novel ATR inhibitor guided by a pharmacophore model to position a key hydrogen bond. Optimization was driven by potency and selectivity over the related kinase mTOR, resulting in the identification of camonsertib (RP-3500) with high potency and excellent ADME properties. Preclinical evaluation focused on the impact of camonsertib on myelosuppression, and an exploration of intermittent dosing schedules to allow recovery of the erythroid compartment and mitigate anemia. Camonsertib is currently undergoing clinical evaluation both as a single agent and in combination with talazoparib, olaparib, niraparib, lunresertib, or gemcitabine (NCT04497116, NCT04972110, NCT04855656). A preliminary recommended phase 2 dose for monotherapy was identified as 160 mg QD given 3 days/week.
Assuntos
Neoplasias , Humanos , Proteínas Mutadas de Ataxia Telangiectasia , Neoplasias/tratamento farmacológico , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/uso terapêutico , GencitabinaRESUMO
A weak antagonist of the pyrimidinergic receptor P2Y(14) containing a dihydropyridopyrimidine core was identified through high-throughput screening. Subsequent optimization led to potent, non-UTP competitive antagonists and represent the first reported non-nucleotide antagonists of this receptor. Compound 18q was identified as a 10 nM P2Y(14) antagonist with good oral bioavailability and provided sufficient exposure in mice to be used as a tool for future in vivo studies.
Assuntos
Antagonistas do Receptor Purinérgico P2/síntese química , Pirimidinas/síntese química , Receptores Purinérgicos P2/química , Administração Oral , Animais , Disponibilidade Biológica , Camundongos , Estrutura Molecular , Pan troglodytes , Antagonistas do Receptor Purinérgico P2/química , Pirimidinas/administração & dosagem , Pirimidinas/química , Receptores Purinérgicos P2Y , Relação Estrutura-AtividadeRESUMO
The trifluoroethylamine group found in cathepsin K inhibitors like odanacatib can be replaced by a difluoroethylamine group. This change increased the basicity of the nitrogen which positively impacted the log D. This translated into an improved oral bioavailability in pre-clinical species. Difluoroethylamine compounds exhibit a similar potency against cathepsin K and selectivity profile against other cathepsins when compared to trifluoroethylamine analogs.
Assuntos
Catepsina K/antagonistas & inibidores , Etilaminas/química , Inibidores de Proteases/química , Administração Oral , Amidas/química , Animais , Compostos de Bifenilo/química , Compostos de Bifenilo/farmacologia , Catepsina K/metabolismo , Cães , Etilaminas/síntese química , Etilaminas/farmacocinética , Inibidores de Proteases/síntese química , Inibidores de Proteases/farmacocinética , RatosRESUMO
In this manuscript we wish to report the discovery of MK-7246 (4), a potent and selective CRTH2 (DP2) antagonist. SAR studies leading to MK-7246 along with two synthetic sequences enabling the preparation of this novel class of CRTH2 antagonist are reported. Finally, the pharmacokinetic and metabolic profile of MK-7246 is disclosed.
Assuntos
Carbolinas/química , Pneumopatias/tratamento farmacológico , Receptores Imunológicos/antagonistas & inibidores , Receptores de Prostaglandina/antagonistas & inibidores , Animais , Carbolinas/farmacocinética , Carbolinas/uso terapêutico , Humanos , Macaca mulatta , Microssomos Hepáticos/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores Imunológicos/metabolismo , Receptores de Prostaglandina/metabolismo , Relação Estrutura-AtividadeRESUMO
MK-0674 is a potent and selective cathepsin K inhibitor from the same structural class as odanacatib with a comparable inhibitory potency profile against Cat K. It is orally bioavailable and exhibits long half-life in pre-clinical species. In vivo studies using deuterated MK-0674 show stereoselective epimerization of the alcohol stereocenter via an oxidation/reduction cycle. From in vitro incubations, two metabolites could be identified: the hydroxyleucine and the glucuronide conjugate which were confirmed using authentic synthetic standards.
Assuntos
Compostos de Bifenilo/administração & dosagem , Compostos de Bifenilo/farmacocinética , Catepsina K/antagonistas & inibidores , Inibidores de Cisteína Proteinase/administração & dosagem , Inibidores de Cisteína Proteinase/farmacocinética , Descoberta de Drogas/métodos , Administração Oral , Animais , Disponibilidade Biológica , Compostos de Bifenilo/química , Catepsina K/metabolismo , Inibidores de Cisteína Proteinase/química , Cães , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Humanos , Macaca mulatta , Coelhos , RatosRESUMO
Odanacatib is a potent, selective, and neutral cathepsin K inhibitor which was developed to address the metabolic liabilities of the Cat K inhibitor L-873724. Substituting P1 and modifying the P2 side chain led to a metabolically robust inhibitor with a long half-life in preclinical species. Odanacatib was more selective in whole cell assays than the published Cat K inhibitors balicatib and relacatib. Evaluation in dermal fibroblast culture showed minimal intracellular collagen accumulation relative to less selective Cat K inhibitors.
Assuntos
Compostos de Bifenilo/farmacologia , Catepsinas/antagonistas & inibidores , Inibidores de Cisteína Proteinase/farmacologia , Animais , Azepinas/química , Azepinas/farmacologia , Catepsina K , Colágeno/efeitos dos fármacos , Colágeno/imunologia , Cães , Fibroblastos/efeitos dos fármacos , Humanos , Modelos Biológicos , Estrutura Molecular , Osteoporose Pós-Menopausa/tratamento farmacológico , Pele/citologia , Relação Estrutura-Atividade , Sulfonas/química , Sulfonas/farmacologiaRESUMO
Identification, characterization and structure elucidation of human metabolites of drug candidates is crucial for the pharmaceutical industry to assess their activity against the therapeutic target of interest and potential toxicological effects. It often requires in vitro synthesis of microgram quantities of metabolites of interest with enzymatic preparations, pre-concentration of the reaction mixture by solid phase extraction (SPE), metabolite isolation using HPLC systems coupled to fraction collectors prior to nuclear magnetic resonance characterization. The method reported herein is a rapid and simple technique using solely off-line mixed phase anionic exchange lipophilic SPE cartridges to selectively isolate glucuronide and sulfate metabolites from their parent compound. This approach capitalizes on the pKa differences between the parent compound, devoided of acidic moieties, and the negatively charged glucuronide and/or sulfate metabolites. Once loaded on the SPE cartridge, the incubation mixture is washed successively with a basic aqueous solution, methanol to elute the non-anionic parent compounds, and then with an acidic methanolic solution to protonate and recover the phase II conjugates. Over 100 microg (>95% purity) of 17 alpha-ethynylestradiol-3-glucuronide and 6-gingerol-4'-glucuronide were successfully isolated using this technique, as well as glucuronide and a sulfate conjugates of 1-{4'-[(1R)-2,2-difluoro-1-hydroxyethyl]biphenyl-4-yl}cyclopropanecarboxamide (DHBC) synthesized in-house. Their structures were confirmed by Ultra Performance Liquid Chromatography coupled to Quadrupole-Time of flight (UPLC-QTof) and nuclear magnetic resonance analysis.
Assuntos
Ânions/química , Cromatografia por Troca Iônica/instrumentação , Desintoxicação Metabólica Fase II , Extração em Fase Sólida/métodos , Espectrometria de Massas por Ionização por Electrospray/métodos , Animais , Cromatografia Líquida de Alta Pressão/instrumentação , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia por Troca Iônica/métodos , Glucuronídeos/química , Humanos , Interações Hidrofóbicas e Hidrofílicas , Mucosa Intestinal/metabolismo , Intestinos/química , Espectroscopia de Ressonância Magnética/métodos , Microssomos Hepáticos/química , Microssomos Hepáticos/metabolismo , Ratos , Ratos Sprague-Dawley , Sensibilidade e Especificidade , Extração em Fase Sólida/instrumentação , Solventes/química , Espectrometria de Massas por Ionização por Electrospray/instrumentação , Espectrometria de Massas em Tandem/métodosRESUMO
Condensation of N-tert-butanesulfinamide (S)-1 with trifluoroacetaldehyde hydrate 2a afforded 2-methyl-N-(2,2,2-trifluoroethylidene)propane-2-sulfinamide 3. Without isolation and purification, imine 3 was added to various aryllithium reagents to give highly diastereomerically enriched adducts 5a-g. Acidic methanolysis of 5a-g provided the desired 1-aryl-2,2,2-trifluoroethylamine hydrochloride compounds 6a-g. [reaction: see text].
Assuntos
Etilaminas/síntese química , Hidrocarbonetos Fluorados/síntese química , Compostos de Sulfônio/química , Cromatografia Líquida de Alta Pressão , Hidrocarbonetos Fluorados/química , Iminas/síntese química , Indicadores e Reagentes , Compostos Organometálicos/química , EstereoisomerismoRESUMO
A novel HIV protease inhibitor was designed using a morpholine core as the aspartate binding group. Analysis of the crystal structure of the initial lead bound to HIV protease enabled optimization of enzyme potency and antiviral activity. This afforded a series of potent orally bioavailable inhibitors of which MK-8718 was identified as a compound with a favorable overall profile.
RESUMO
[reaction: see text] The use of anionic polycyclization (AP) in constructing the steroidal backbone of cardenolides was investigated. The reaction of 2-carbomethoxy-2-cyclohexenone I with the enolate of Nazarov reagent II gave, after decarboxylation and aldol condensation, steroid III with control of stereochemistry.
Assuntos
Cardiotônicos/síntese química , Esteroides/síntese química , Cardenolídeos/síntese química , Ciclização , Ouabaína/síntese química , EstereoisomerismoRESUMO
Highly potent, selective, and bioavailable inhibitors of human, mouse, or rat cathepsin S are described. The key structural features combine a sulfonyl moiety attached to a large group in P2 and a small substituent in P3.
Assuntos
Catepsinas/antagonistas & inibidores , Química Farmacêutica/métodos , Animais , Inibidores de Cisteína Proteinase/química , Desenho de Fármacos , Avaliação Pré-Clínica de Medicamentos , Humanos , Concentração Inibidora 50 , Camundongos , Modelos Químicos , Modelos Moleculares , Conformação Molecular , Estrutura Molecular , Ligação Proteica , Ratos , Relação Estrutura-AtividadeRESUMO
Based on our previous study with trifluoroethylamine as a P2-P3 amide isostere of cathepsin K inhibitor, further optimization led to identification of compound 22 (L-873724) as a potent and selective non-basic cathepsin K inhibitor. This compound showed excellent pharmacokinetics and efficacy in an ovariectomized (OVX) rhesus monkey model. The volumes of distribution close to unity were consistent with this compound not being lysosomotropic, which is a characteristic of basic cathepsin K inhibitors.