RESUMO
An application of Fourier transform infrared (FT-IR) spectroscopy equipped with an attenuated total reflectance (ATR) probe for in-line monitoring of a hydrochloride (HCl) salt formation process of 4-{1-methyl-2-piperidin-4-yl-4-[3-(trifluorometryl)phenyl]-1H-imidazol-5-yl}-N-[(1S)-1-phenylethyl]pyridine-2-amine (freebase), an active pharmaceutical ingredient as a P38 MAP kinase inhibitor, is described. The freebase forms both mono- and bis-HCl salts due to its structural features. The mono-HCl salt is the desired product but the bis-salt is an impurity. The key to maximizing the product yield and minimizing the impurity level is to monitor the salt-forming reaction and to terminate it at the correct HCl charge amount. The process analytical technology (PAT) provided real-time data for process control and overcame the limitations that had been previously encountered by other analytical instrumentations, such as high-performance liquid chromatography and titration. Two qualitative approaches for reaction endpoint determination were employed. In the first approach, changes in the concentration of the freebase and bis-salt were monitored via the first derivative concentration profiles. The flat point in the freebase profile and the rise in the bis-salt profile were used as a detection bracket for the endpoint of HCl charging. In the second approach, principal component analysis (PCA) was used to classify the status of the process based on a spectral library consisting of spectra collected around the endpoint. Results indicated that both methods provided adequate accuracy for endpoint control in a small window between 1.0 and 1.05 HCl to freebase mole ratio. Both methods were used to support a scaled up process. Three batches of MAP mono-HCl salt formation were successfully controlled and prepared.
Assuntos
Química Farmacêutica/métodos , Preparações Farmacêuticas/análise , Sais/química , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Tecnologia Farmacêutica/métodos , Calibragem , Cromatografia Líquida de Alta Pressão , Modelos Químicos , Análise de Componente Principal , Sais/análise , Proteínas Quinases p38 Ativadas por Mitógeno/análiseRESUMO
This paper reports a comparison study of the difference between Chiralpak AD-H and AD columns in enantioseparation of dihydropyrimidinone (DHP) acid and its methyl ester under normal phase LC conditions. Unlike those of the AD phase, the van't Hoff plots of retention factors for DHP acid on the AD-H phase were linear. The cyclic van't Hoff plots of selectivity factors for DHP acid on the AD-H phase were non-linear and slightly non-superimposable. No conformational transition was observed on the AD-H phase in the whole temperature range. A single-step temperature program on the AD-H phase showed that the selectivity factors of DHP acid only increased approximately 1.7% in 24 h (versus approximately 50% on the AD phase). For DHP ester, the single-step temperature program showed that the selectivity factors on the AD-H phase remained the same in 24 h while those on the AD phase increased around 3.1%. The enantioselectivity of DHP acid on the AD-H phase was lower than that on the AD phase while the enantioselectivity of DHP ester on the AD-H phase was higher than that on the AD phase. The resolution of DHP acid on the AD-H phase was about the same as that on the AD phase while the resolution of DHP ester on the AD-H phase was much higher than that on the AD phase. The results of DHP acid are opposite of what the vendor suggested while the results of DHP ester are the same as the vendor's application notes. This indicates that the differences between Chiralpak AD-H and AD columns are not only in their particle size, but also in the solvated conformations.
Assuntos
Amilose/análogos & derivados , Amilose/química , Ésteres/isolamento & purificação , Fenilcarbamatos/química , Pirimidinonas/isolamento & purificação , Pirimidinonas/química , Estereoisomerismo , TemperaturaRESUMO
The possible mechanisms for the chiral recognition of 2(S)-(3,5-bis-trifluoromethyl-phenyl)-2-[3(S)-(4-fluorophenyl)-4-(1H-[1,2,4]triazol-3-ylmethyl)-morpholin-2(R)-yloxy]-ethanol (compound A) and its enantiomer with native gamma-cyclodextrin (gamma-CD) were investigated using capillary electrophoresis (CE), reversed-phase liquid chromatography (RPLC), proton (1H), fluorine (19F) and carbon (13C) nuclear magnetic resonance spectroscopy (NMR), electrospray mass spectrometry (ESI-MS) and circular dichroism (CD). All experiments provided clear evidence of the formation of diastereomeric complexes between the enantiomers and gamma-CD. Proton, fluorine and carbon NMR spectra suggested that both aromatic rings, with mono-fluoro and bis-tri-fluoro functional groups, on the guest molecule were partially included into the cavity of the gamma-CD. ESI-MS spectra indicated that the diastereomeric complexes have a 1:1 stoichiometric ratio. The binding constants of the diastereomeric complexes obtained by CE, RPLC and CD were compared. The effects of the gamma-CD concentration, organic modifiers and temperature on the CE-chiral separation were also investigated.
Assuntos
Cromatografia Líquida/métodos , Dicroísmo Circular/métodos , Ciclodextrinas/química , Eletroforese Capilar/métodos , Espectroscopia de Ressonância Magnética/métodos , Espectrometria de Massas por Ionização por Electrospray/métodos , gama-Ciclodextrinas , EstereoisomerismoRESUMO
Enantioseparations of basic pharmaceutical compounds were investigated using different types of sulfated cyclodextrins as chiral selectors. A general strategy for method development was described, together with enantiomeric separations of a number of pharmaceutical related compounds. Based on this strategy, systematic method development approaches for several selected compounds were performed by modifying method parameters, such as the concentration of the chiral selectors, buffer pH, type of organic modifiers, buffer type, temperature and applied voltage. The results of the investigation elucidated the separation mechanism. Many practical aspects were also discussed through several specific examples in order to demonstrate how to develop and validate a precise, sensitive, accurate and rugged separation.
Assuntos
Ciclodextrinas/análise , Sulfatos/análise , Ciclodextrinas/química , Eletroforese Capilar/métodos , Preparações Farmacêuticas/análise , Preparações Farmacêuticas/química , Estereoisomerismo , Sulfatos/químicaRESUMO
Rofecoxib is a highly active and selective cyclo-oxygenase II inhibitor. A stability-indicating method for the assay of rofecoxib has been developed using reverse-phase high-performance liquid chromatography (HPLC). Stress testing of rofecoxib was conducted during the method development and validation. HPLC analysis of rofecoxib solutions stressed under alkaline and photolytic conditions revealed the presence of several degradates. Two main degradates were determined to be the cyclization product formed by photo-cyclization and the dicarboxylate formed by ring opening in the presence of base and oxygen. The identities of these degradates were confirmed by comparison of UV spectra and HPLC retention time with the independently synthesized products. The mechanistic pathways for the formation of these degradates are discussed. Further improvement of the HPLC method's ruggedness has been made based on these studies.
Assuntos
Anti-Inflamatórios não Esteroides/química , Estabilidade de Medicamentos , Lactonas/química , Fotólise , Álcalis , Cromatografia Líquida de Alta Pressão , Concentração de Íons de Hidrogênio , Soluções , SulfonasRESUMO
Due to the high method variability (typically > or = 0.5%, based on a literature survey and internal Merck experience) encountered in the HPLC weight percent (%) assays of various active pharmaceutical ingredients (APIs), it is proposed that the routine use of the test in stability studies should be discouraged on the basis that it is frequently not sufficiently precise to yield results that are stability-indicating. The high method variability of HPLC weight % methods is not consistent with the current ICH practice of reporting impurities/degradation products down to the 0.05% level, and it can lead to erroneous out-of-specification (OOS) results that are due to experimental error and are not attributable to API degradation. For the vast majority of cases, the HPLC impurity profile provides much better (earlier and more sensitive) detection of low-level degradation products. Based on these observations, a Quality-by-Design (QbD) approach is proposed to phase out the HPLC weight % assay from routine API stability testing protocols.
Assuntos
Química Farmacêutica/métodos , Química Farmacêutica/normas , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia Líquida de Alta Pressão/normas , Estabilidade de Medicamentos , Composição de Medicamentos , Contaminação de Medicamentos , Indústria Farmacêutica , Preparações Farmacêuticas/análise , Controle de Qualidade , Reprodutibilidade dos Testes , Tecnologia Farmacêutica/métodos , Tecnologia Farmacêutica/normasRESUMO
A novel approach for the potential on-line determination of the enantiomeric excess (ee) of an M3 antagonist drug substance combining attenuated total reflectance infrared (ATR-IR) spectroscopy, guest-host complexes, and chemometric data analysis is described. Chiral recognition through a formation of diastereomeric complexes was measured by ATR-IR. Small changes on the IR spectra reflect the interaction between the guest (M3) and host (chiral selector). These changes are measured as a function of M3 enantiomer excess. The standard error of prediction is 1.3 ee%. The prediction results based on the IR method were in good agreement with the gravimetric method. The robustness of the calibration model was evaluated by varying the concentration of the chiral selector, the pH of the solution, and the organic solvents. The stability of the calibration model was also demonstrated through measuring different sets of samples on different days.
Assuntos
1-(5-Isoquinolinasulfonil)-2-Metilpiperazina/análogos & derivados , Ciclodextrinas/análise , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Tartaratos/análise , 1-(5-Isoquinolinasulfonil)-2-Metilpiperazina/antagonistas & inibidores , 1-(5-Isoquinolinasulfonil)-2-Metilpiperazina/química , 2-Propanol/química , Acetonitrilas/química , Calibragem , Ciclodextrinas/química , Análise dos Mínimos Quadrados , Metanol/química , Estrutura Molecular , Solventes/química , Estereoisomerismo , Tartaratos/químicaRESUMO
The synthesis of pharmaceutical products frequently involves the use of reactive reagents and the formation of intermediates and by-products. Low levels of some of these may be present in the final drug substance and drug product as impurities. Such chemically reactive impurities may have at the same time the potential for unwanted toxicities including genotoxicity and carcinogenicity and hence can have an impact on product risk assessment. This paper outlines a procedure for testing, classification, qualification, toxicological risk assessment, and control of impurities possessing genotoxic potential in pharmaceutical products. Referencing accepted principles of cancer risk assessment, this document proposes a staged threshold of toxicological concern (TTC) approach for the intake of genotoxic impurities over various periods of exposure. This staged TTC is based on knowledge about tumorigenic potency of a wide range of genotoxic carcinogens and can be used for genotoxic compounds, for which cancer data are limited or not available. The delineated acceptable daily intake values of between approximately 1.5 microg/day for approximately lifetime intake and approximately 120 microg/day for < or = 1 month are virtually safe doses. Based on sound scientific reasoning, these virtually safe intake values do not pose an unacceptable risk to either human volunteers or patients at any stage of clinical development and marketing of a pharmaceutical product. The intake levels are estimated to give an excess cancer risk of 1 in 100,000 to 1 in a million over a lifetime, and are extremely conservative given the current lifetime cancer risk in the population of over 1 in 4 (http://seer.cancer.gov/statfacts/html.all.html). The proposals in this document apply to all clinical routes of administration and to compounds at all stages of clinical development. It is important to note that certain types of products, such as those for life-threatening indications for which there are no safer alternatives, allow for special considerations using adaptations of the principles outlined in this paper.
Assuntos
Contaminação de Medicamentos/prevenção & controle , Mutagênicos/análise , Preparações Farmacêuticas/síntese química , Animais , Carcinógenos/análise , Carcinógenos/química , Carcinógenos/classificação , Guias como Assunto , Humanos , Mutagênicos/química , Mutagênicos/classificação , Medição de Risco/métodos , Relação Estrutura-AtividadeRESUMO
The chiral separation of a substituted imidazole p38 MAP kinase inhibitor and its intermediates was investigated using capillary electrophoresis (CE) with various sulfated cyclodextrins. After initial screens, a single CE chiral method with a randomly sulfated beta-CD was selected for the evaluation of chiral purity for all three compounds. Operational parameters, such as the concentration of the chiral selectors, background electrolyte (or mobile phase) pH, organic modifiers, and temperature were varied in order to achieve an optimized method. The optimal method was validated in terms of linearity, sensitivity, precision, ruggedness, and specificity.
Assuntos
Eletroforese Capilar/métodos , Inibidores Enzimáticos/química , Inibidores Enzimáticos/isolamento & purificação , Imidazóis/química , Imidazóis/isolamento & purificação , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Acetonitrilas , Concentração de Íons de Hidrogênio , Indicadores e Reagentes , Estrutura Molecular , Soluções , Estereoisomerismo , Temperatura , beta-Ciclodextrinas/químicaRESUMO
The possible mechanisms for the chiral recognition of 2-(R)-N-[1-(6-aminopyridin-2-ylmethyl)piperidin-4-yl]-2-[(1R)-3,3-difluorocyclopentyl]-2-hydroxy-2-phenylacetamide (RR-M3), and its enantiomer (SS-M3) with octakis(2,3-di-O-acetyl-6-sulfo)-gamma-cyclodextrin (ODAS-gamma-CD) and octakis(6-sulfo)-gamma-cycöpdextrom enantiomer; (OS-gamma-CD), were investigated using capillary electrophoresis (CE), proton ((1)H), fluorine ((19)F) and carbon ((13)C) nuclear magnetic resonance spectroscopy (NMR), and infrared (IR) spectroscopy. Clear evidence for the formation of diastereomeric complexes between the enantiomers and the two CDs was observed. NMR spectra suggest that the phenyl and difluorocyclopentyl rings are involved in the complexation. The phenyl ring on the guest molecule is deeply penetrated into the cavity of OS-gamma-CD, but it is not included into the cavity of ODAS-gamma-CD. The continuous variation plots built based on the (1)H NMR and IR spectra indicate a 1:1 complex stoichiometric ratio of the M3 enantiomers for both CDs. The affinity of the enantiomers for the two CDs is opposite.