Plasma and Liver Protein Binding of N-Acetylgalactosamine-Conjugated Small Interfering RNA.
Drug Metab Dispos
; 47(10): 1174-1182, 2019 10.
Article
em En
| MEDLINE
| ID: mdl-31097425
ABSTRACT
Understanding small interfering RNA (siRNA) fraction unbound (f u) in relevant physiologic compartments is critical for establishing pharmacokinetic-pharmacodynamic relationships for this emerging modality. In our attempts to isolate the equilibrium free fraction of N-acetylgalactosamine-conjugated siRNA using classic small-molecule in vitro techniques, we found that the hydrodynamic radius was critical in determining the size exclusion limit requirements for f u isolation, largely validating the siRNA "rigid rod" hypothesis. With this knowledge, we developed an orthogonally validated 50 kDa molecular-mass cutoff ultrafiltration assay to quantify f u in biologic matrices including human, nonhuman primate, rat, and mouse plasma, and human liver homogenate. To enhance understanding of the siRNA-plasma interaction landscape, we examined the effects of various common oligonucleotide therapeutic modifications to the ribose and helix backbone on siRNA f u in plasma (f u,plasma) and found that chemical modifications can alter plasma protein binding by at least 20%. Finally, to gain insight into which specific plasma proteins bind to siRNA, we developed a qualitative screen to identify binding "hits" across a panel of select purified human plasma proteins.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Acetilgalactosamina
/
Proteínas Sanguíneas
/
RNA Interferente Pequeno
/
Fígado
Tipo de estudo:
Qualitative_research
Limite:
Adult
/
Animals
/
Female
/
Humans
Idioma:
En
Revista:
Drug Metab Dispos
Ano de publicação:
2019
Tipo de documento:
Article