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Utility of Cellular Measurements of Non-Specific Endocytosis to Assess the Target-Independent Clearance of Monoclonal Antibodies.
Bryniarski, Mark A; Tuhin, Md Tariqul Haque; Shomin, Carolyn D; Nasrollahi, Fatemeh; Ko, Eunkyung Clare; Soto, Marcus; Chung, Kyu; Poon-Andersen, Carrie; Primack, Ronya; Wong, Diana; Ojeda, Esperanza; Chung, John; Cook, Kevin D; Conner, Kip P.
Afiliación
  • Bryniarski MA; Department of Pharmacokinetics and Drug Metabolism, Amgen Inc., 750 Gateway Blvd, Suite 100, South San Francisco, CA, 94080, USA. Electronic address: mbryniar@amgen.com.
  • Tuhin MTH; Department of Pharmacokinetics and Drug Metabolism, Amgen Inc., 750 Gateway Blvd, Suite 100, South San Francisco, CA, 94080, USA.
  • Shomin CD; Department of Biologics, Amgen Inc., 1 Amgen Center Drive, Thousand Oaks, CA, 91320, USA.
  • Nasrollahi F; Process Development; Pre-Pivotal Drug Product Technologies, Amgen Inc., 1 Amgen Center Drive, Thousand Oaks, CA, 91320, USA.
  • Ko EC; Department of Pharmacokinetics and Drug Metabolism, Amgen Inc., 750 Gateway Blvd, Suite 100, South San Francisco, CA, 94080, USA.
  • Soto M; Pharmacokinetics & Drug Metabolism, Amgen Research, 1 Amgen Center Drive, Thousand Oaks, CA, 91320, USA.
  • Chung K; Department of Pharmacokinetics and Drug Metabolism, Amgen Inc., 750 Gateway Blvd, Suite 100, South San Francisco, CA, 94080, USA.
  • Poon-Andersen C; Department of Pharmacokinetics and Drug Metabolism, Amgen Inc., 750 Gateway Blvd, Suite 100, South San Francisco, CA, 94080, USA.
  • Primack R; Pharmacokinetics & Drug Metabolism, Amgen Research, 1 Amgen Center Drive, Thousand Oaks, CA, 91320, USA.
  • Wong D; Department of Pharmacokinetics and Drug Metabolism, Amgen Inc., 750 Gateway Blvd, Suite 100, South San Francisco, CA, 94080, USA.
  • Ojeda E; Pharmacokinetics & Drug Metabolism, Amgen Research, 1 Amgen Center Drive, Thousand Oaks, CA, 91320, USA.
  • Chung J; Process Development; Pre-Pivotal Drug Product Technologies, Amgen Inc., 1 Amgen Center Drive, Thousand Oaks, CA, 91320, USA.
  • Cook KD; Department of Pharmacokinetics and Drug Metabolism, Amgen Inc., 750 Gateway Blvd, Suite 100, South San Francisco, CA, 94080, USA.
  • Conner KP; Department of Pharmacokinetics and Drug Metabolism, Amgen Inc., 750 Gateway Blvd, Suite 100, South San Francisco, CA, 94080, USA. Electronic address: kipc@amgen.com.
J Pharm Sci ; 113(10): 3100-3111, 2024 Oct.
Article en En | MEDLINE | ID: mdl-39009346
ABSTRACT
Past studies have demonstrated higher clearance for monoclonal antibodies possessing increased rates of non-specific endocytosis. However, this metric is oftentimes evaluated indirectly using biophysical techniques or cell surface binding studies that may not provide insight into the specific rates of cellular turnover. Furthermore, few examples evaluating non-specific endocytosis have been reported for a therapeutic antibody that reached clinical assessment. In the current report, we evaluated a therapeutic human immunoglobulin G2 monoclonal antibody targeted against the interleukin-4 receptor alpha chain (IL-4Rα) that exhibited elevated target independent clearance in previous Phase 1 and 2 studies. We confirmed high non-specific clearance of the anti-IL-4Rα antibody as compared to a reference antibody during pharmacokinetic assessments in wild type mice where target-mediated disposition was absent. We then developed a cell-based method capable of measuring cellular protein endocytosis and demonstrated the anti-IL-4Rα antibody exhibited marked non-specific uptake relative to the reference compound. Antibody homology modeling identified the anti-IL-4Rα antibody possessed positive charge patches whose removal via targeted mutations substantially reduced its non-specific endocytosis. We then expanded the scope of the study by evaluating panels of both preclinical and clinically relevant monoclonal antibodies and demonstrate those with the highest rates of non-specific uptake in vitro exhibited elevated target independent clearance, low subcutaneous bioavailability, or both. Our results support the observation that high non-specific endocytosis is a negative attribute in monoclonal antibody development and demonstrate the utility of a generic cell-based screen as a quantitative tool to measure non-specific endocytosis of protein therapeutics at the single-cell level.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Endocitosis / Anticuerpos Monoclonales Límite: Animals / Humans / Male Idioma: En Revista: J Pharm Sci Año: 2024 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Endocitosis / Anticuerpos Monoclonales Límite: Animals / Humans / Male Idioma: En Revista: J Pharm Sci Año: 2024 Tipo del documento: Article Pais de publicación: Estados Unidos