Design and preclinical characterization of a novel vaccine adjuvant formulation consisting of a synthetic TLR4 agonist in a thermoreversible squalene emulsion

Int J Pharm. 2015;486(1-2):99-111. doi: 10.1016/j.ijpharm.2015.03.028. Epub 2015 Mar 17.

Abstract

We describe the development, analytical characterization, stability and preclinical efficacy of AF04, a combination adjuvant comprising the synthetic toll-like receptor 4 (TLR4) agonist, E6020, formulated in AF03, a thermoreversible squalene emulsion. By using AF04 with the recombinant major outer membrane protein of Chlamydia trachomatis (Ct-MOMP) and with the recombinant surface glycoprotein gB from human cytomegalovirus (CMV-gB) as model antigens, we show that AF03 and E6020 can synergize to augment specific antibody and Th-1 cellular immune responses in mice. In terms of formulation, we observe that the method used to incorporate E6020 into AF03 affects its partition between the oil and water phases of the emulsion which in turn has a significant impact on the tolerability (IV pyrogenicity test in rabbits) of this novel adjuvant combination.

Keywords: AF03; AF04; Adjuvant; CMV; Chlamydia trachomatis; E6020.

MeSH terms

  • Adjuvants, Immunologic* / chemistry
  • Adjuvants, Immunologic* / pharmacology
  • Animals
  • Antibodies, Bacterial / blood
  • Antibodies, Viral / blood
  • Antigens, Bacterial / immunology
  • Antigens, Viral / immunology
  • Bacterial Outer Membrane Proteins / immunology*
  • Cell Line
  • Chlamydia trachomatis
  • Cytokines / immunology
  • Cytomegalovirus
  • Dendritic Cells / drug effects
  • Dendritic Cells / immunology
  • Drug Design
  • Emulsions
  • Female
  • Glycoproteins / immunology
  • Humans
  • Immunoglobulin G / blood
  • Leukocytes, Mononuclear
  • Mice, Inbred C57BL
  • Rabbits
  • Squalene* / chemistry
  • Squalene* / pharmacology
  • Toll-Like Receptor 4 / agonists*
  • Vaccines* / chemistry
  • Vaccines* / pharmacology
  • Viral Proteins / immunology*

Substances

  • Adjuvants, Immunologic
  • Antibodies, Bacterial
  • Antibodies, Viral
  • Antigens, Bacterial
  • Antigens, Viral
  • Bacterial Outer Membrane Proteins
  • Cytokines
  • Emulsions
  • Glycoproteins
  • Immunoglobulin G
  • Tlr4 protein, mouse
  • Toll-Like Receptor 4
  • Vaccines
  • Viral Proteins
  • Squalene