The development and optimisation of an HPLC-based in vitro serum stability assay for a calcitonin gene-related peptide receptor antagonist peptide.
J Pept Sci
; 30(2): e3539, 2024 Feb.
Article
em En
| MEDLINE
| ID: mdl-37605343
ABSTRACT
Evaluation of the stability of peptide drug candidates in biological fluids, such as blood serum, is of high importance during the lead optimisation phase. Here, we describe the optimisation and validation of a method for the evaluation of the stability of a lead calcitonin gene-related peptide antagonist peptide (P006) in blood serum. After initially determining appropriate peptide and human serum concentrations and selection of the quenching reagent, the HPLC method optimisation used two experimental designs, Plackett-Burman design and Taguchi design. The analytical method was validated as complying with the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use guidelines. The optimised method allowed the successful resolution of the parent peptide from its metabolites using RP-HPLC and identification of the major metabolites of P006 by mass spectrometry. This paradigm may be widely adopted as a robust early-stage platform for screening peptide stability to rule out candidates with low in vitro stability, which would likely translate into poor in vivo pharmacokinetics.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Peptídeo Relacionado com Gene de Calcitonina
/
Antagonistas do Receptor do Peptídeo Relacionado ao Gene de Calcitonina
Limite:
Humans
Idioma:
En
Revista:
J Pept Sci
Assunto da revista:
BIOQUIMICA
Ano de publicação:
2024
Tipo de documento:
Article
País de afiliação:
Reino Unido