Effects of altered FcγR binding on antibody pharmacokinetics in cynomolgus monkeys.
Antibody interactions with Fcγ receptors (FcγRs), like FcγRIIIA, play a critical role in mediating antibody effector functions and thereby contribute significantly to the biologic and therapeutic activity of antibodies. Over the past decade, considerable work has been directed towards production of antibodies with altered binding affinity to FcγRs and evaluation of how the alterations modulate their therapeutic activity. This has been achieved by altering glycosylation status at N297 or by engineering modifications in the crystallizable fragment (Fc) region. While the effects of these modifications on biologic activity and efficacy have been examined, few studies have been conducted to understand their effect on antibody pharmacokinetics (PK). We present here a retrospective analysis in which we characterize the PK of three antibody variants with decreased FcγR binding affinity caused by amino acid substitutions in the Fc region (N297A, N297G, and L234A/L235A) and three antibody variants with increased FcγRIIIA binding affinity caused by afucosylation at N297, and compare their PK to corresponding wild type antibody PK in cynomolgus monkeys. For all antibodies, PK was examined at a dose that was known to be in the linear range. Since production of the N297A and N297G variants in Chinese hamster ovary cells results in aglycosylated antibodies that do not bind to FcγRs, we also examined the effect of expression of an aglycosylated antibody, without sequence change(s), in E. coli. All the variants demonstrated similar PK compared with that of the wild type antibodies, suggesting that, for the six antibodies presented here, altered FcγR binding affinity does not affect PK.
Department of Pharmacokinetics and Pharmacodynamics; Genentech, Inc; San Francisco, CA USA.
Maya K
MK
Leabman
Department of Biochemical and Cellular Pharmacology; Genentech, Inc; San Francisco, CA USA.
Y Gloria
YG
Meng
Department of Antibody Engineering; Genentech, Inc; San Francisco, CA USA.
Robert F
RF
Kelley
Department of Biochemical and Cellular Pharmacology; Genentech, Inc; San Francisco, CA USA.
Laura E
LE
DeForge
Department of BioAnalytical Sciences; Genentech, Inc; San Francisco, CA USA.
Kyra J
KJ
Cowan
Department of Pharmacokinetics and Pharmacodynamics; Genentech, Inc; San Francisco, CA USA.
Suhasini
S
Iyer
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000378:metabolism / Q000493:pharmacokinetics*; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D014644:Genetic Variation; D008252:Macaca fascicularis; D011485:Protein Binding; D011961:Receptors, Fc / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D011961:Receptors, Fc
FcγR binding; afucosylation; aglycosylation; antibody; pharmacokinetics