A PCSK9-binding antibody that structurally mimics the EGF(A) domain of LDL-receptor reduces LDL cholesterol in vivo

J Lipid Res. 2011 Jan;52(1):78-86. doi: 10.1194/jlr.M011445. Epub 2010 Oct 19.

Abstract

Proprotein convertase subtilisin-like/kexin type 9 (PCSK9) regulates LDL cholesterol levels by inhibiting LDL receptor (LDLr)-mediated cellular LDL uptake. We have identified a fragment antigen-binding (Fab) 1D05 which binds PCSK9 with nanomolar affinity. The fully human antibody 1D05-IgG2 completely blocks the inhibitory effects of wild-type PCSK9 and two gain-of-function human PCSK9 mutants, S127R and D374Y. The crystal structure of 1D05-Fab bound to PCSK9 reveals that 1D05-Fab binds to an epitope on the PCSK9 catalytic domain which includes the entire LDLr EGF(A) binding site. Notably, the 1D05-Fab CDR-H3 and CDR-H2 loops structurally mimic the EGF(A) domain of LDLr. In a transgenic mouse model (CETP/LDLr-hemi), in which plasma lipid and PCSK9 profiles are comparable to those of humans, 1D05-IgG2 reduces plasma LDL cholesterol to 40% and raises hepatic LDLr protein levels approximately fivefold. Similarly, in healthy rhesus monkeys, 1D05-IgG2 effectively reduced LDL cholesterol 20%-50% for over 2 weeks, despite its relatively short terminal half-life (t(1/2) = 3.2 days). Importantly, the decrease in circulating LDL cholesterol corresponds closely to the reduction in free PCSK9 levels. Together these results clearly demonstrate that the LDL-lowering effect of the neutralizing anti-PCSK9 1D05-IgG2 antibody is mediated by reducing the amount of PCSK9 that can bind to the LDLr.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / metabolism
  • Binding Sites
  • Cholesterol Ester Transfer Proteins / metabolism
  • Cholesterol, LDL / blood*
  • Fluoroimmunoassay
  • Humans
  • Immunoglobulin Fab Fragments / chemistry
  • Immunoglobulin Fab Fragments / pharmacology*
  • Immunoglobulin G / immunology
  • Immunoglobulin G / metabolism
  • Macaca mulatta
  • Male
  • Mice
  • Mice, Transgenic
  • Proprotein Convertase 9
  • Proprotein Convertases
  • Receptors, LDL / chemistry*
  • Receptors, LDL / metabolism
  • Serine Endopeptidases / chemistry
  • Serine Endopeptidases / immunology*

Substances

  • Antibodies, Monoclonal
  • CETP protein, human
  • Cholesterol Ester Transfer Proteins
  • Cholesterol, LDL
  • Immunoglobulin Fab Fragments
  • Immunoglobulin G
  • Receptors, LDL
  • PCSK9 protein, human
  • Proprotein Convertase 9
  • Proprotein Convertases
  • Serine Endopeptidases

Associated data

  • PDB/2XTJ