An anti-PCSK9 antibody reduces LDL-cholesterol on top of a statin and suppresses hepatocyte SREBP-regulated genes

Int J Biol Sci. 2012;8(3):310-27. doi: 10.7150/ijbs.3524. Epub 2012 Feb 9.

Abstract

Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a promising therapeutic target for treating coronary heart disease. We report a novel antibody 1B20 that binds to PCSK9 with sub-nanomolar affinity and antagonizes PCSK9 function in-vitro. In CETP/LDLR-hemi mice two successive doses of 1B20, administered 14 days apart at 3 or 10 mpk, induced dose dependent reductions in LDL-cholesterol (≥ 25% for 7-14 days) that correlated well with the extent of PCSK9 occupancy by the antibody. In addition, 1B20 induces increases in total plasma antibody-bound PCSK9 levels and decreases in liver mRNA levels of SREBP-regulated genes PCSK9 and LDLR, with a time course that parallels decreases in plasma LDL-cholesterol (LDL-C). Consistent with this observation in mice, in statin-responsive human primary hepatocytes, 1B20 lowers PCSK9 and LDLR mRNA levels and raises serum steady-state levels of antibody-bound PCSK9. In addition, mRNA levels of several SREBP regulated genes involved in cholesterol and fatty-acid synthesis including ACSS2, FDPS, IDI1, MVD, HMGCR, and CYP51A1 were decreased significantly with antibody treatment of primary human hepatocytes. In rhesus monkeys, subcutaneous (SC) dosing of 1B20 dose-dependently induces robust LDL-C lowering (maximal ~70%), which is correlated with increases in target engagement and total antibody-bound PCSK9 levels. Importantly, a combination of 1B20 and Simvastatin in dyslipidemic rhesus monkeys reduced LDL-C more than either agent alone, consistent with a mechanism of action that predicts additive effects of anti-PCSK9 agents with statins. Our results suggest that antibodies targeting PCSK9 could provide patients powerful LDL lowering efficacy on top of statins, and lower cardiovascular risk.

Keywords: LDL receptor; PCSK9; hypercholesterolemia; low density lipoprotein cholesterol (LDL-C); primary hepatocytes; sterol regulatory element binding protein (SREBP).

MeSH terms

  • Animals
  • Antibodies, Monoclonal / pharmacology
  • Antibodies, Monoclonal / therapeutic use*
  • Antibody Affinity
  • Anticholesteremic Agents / administration & dosage
  • Anticholesteremic Agents / therapeutic use*
  • Cells, Cultured / drug effects
  • Cells, Cultured / metabolism
  • Cholesterol Ester Transfer Proteins / genetics
  • Cholesterol Ester Transfer Proteins / metabolism
  • Cholesterol, LDL / blood*
  • Gene Expression Profiling
  • Gene Expression Regulation / drug effects*
  • Hep G2 Cells / drug effects
  • Hep G2 Cells / metabolism
  • Hepatocytes / drug effects*
  • Hepatocytes / metabolism
  • Humans
  • Immunization, Passive*
  • Lipid Metabolism / genetics
  • Liver / drug effects
  • Liver / metabolism
  • Macaca mulatta
  • Metabolic Syndrome / drug therapy
  • Metabolic Syndrome / genetics
  • Metabolic Syndrome / therapy*
  • Mice
  • Mice, Transgenic
  • Proprotein Convertase 9
  • Proprotein Convertases / antagonists & inhibitors*
  • Proprotein Convertases / biosynthesis
  • Proprotein Convertases / genetics
  • Proprotein Convertases / immunology*
  • RNA, Messenger / metabolism
  • Receptors, LDL / biosynthesis
  • Receptors, LDL / genetics
  • Recombinant Proteins / metabolism
  • Serine Endopeptidases / biosynthesis
  • Serine Endopeptidases / genetics
  • Serine Endopeptidases / immunology*
  • Simvastatin / administration & dosage
  • Simvastatin / therapeutic use*
  • Sterol Regulatory Element Binding Proteins / physiology*

Substances

  • Antibodies, Monoclonal
  • Anticholesteremic Agents
  • CETP protein, human
  • Cholesterol Ester Transfer Proteins
  • Cholesterol, LDL
  • RNA, Messenger
  • Receptors, LDL
  • Recombinant Proteins
  • Sterol Regulatory Element Binding Proteins
  • Simvastatin
  • PCSK9 protein, human
  • Pcsk9 protein, mouse
  • Proprotein Convertase 9
  • Proprotein Convertases
  • Serine Endopeptidases