Pharmaceutical and preclinical evaluation of Advax adjuvant as a dose-sparing strategy for ant venom immunotherapy

Abstract

a b s t r a c tA major challenge in broader clinical application of Jack Jumper ant venom immunotherapy (JJA VIT) is the scarcity of ant venom which needs to be manually harvested from wild ants. Adjuvants are commonly used for antigen sparing in other vaccines, and thereby could potentially have major benefits to extend JJA supplies if they were to similarly enhance JJA VIT immunogenicity. The purpose of this study was to evaluate the physicochemical and microbiological stability and murine immunogenicity of low-dose JJA VIT formulated with a novel polysaccharide adjuvant referred to as delta inulin or Advax TM . Jack Jumper ant venom (JJAV) protein stability was assessed by UPLC-UV, SDS-PAGE, SDS-PAGE immunoblot, and ELISA inhibition. Diffraction light scattering was used to assess particle size distribution of Advax; pH and benzyl alcohol quantification by UPLC-UV were used to assess the physicochemical stability of JJAV diluent, and endotoxin content and preservative efficacy test was used to investigate the microbiological properties of the adjuvanted VIT formulation. To assess the effect of adjuvant on JJA venom immunogenicity, mice were immunised four times with JJAV alone or formulated with Advax adjuvant. JJA VIT formulated with Advax was found to be physicochemically and microbiologically stable for at least 2 days when stored at 4 and 25 • C with a trend for an increase in allergenic potency observed beyond 2 days of storage. Low-dose JJAV formulated with Advax adjuvant induced significantly higher JJAV-specific IgG than a 5-fold higher dose of JJAV alone, consistent with a powerful allergen-sparing effect. The pharmaceutical data provides important guidance on the formulation, storage and use of JJA VIT formulated with Advax adjuvant, with the murine immunogenicity studies providing a strong rationale for a planned clinical trial to test the ability of Advax adjuvant to achieve 4-fold JJAV dose sparing in JJA-allergic human patients.

Evaluation of a Novel Adjuvanted Vaccine for Ultrashort Regimen Therapy of Artemisia Pollen-Induced Allergic Bronchial Asthma in a Mouse Model

“…Advax™ adjuvant has been shown to induce both Th1 and Th2 cell responses, enhancing the production of IgG without increasing IgE ( 38 ). Advax adjuvant has already been tested successfully in preclinical ( 39 ) and clinical studies ( 40 ) of ASIT against bee and ant venom allergy. Advax can be used in combination with a CpG oligonucleotide (Advax-CpG) that stimulates TLR-9, which results in even stronger activation of humoral and cellular immunity ( 41 ).…”

The Peptide Venom Composition of the Fierce Stinging Ant Tetraponera aethiops (Formicidae: Pseudomyrmecinae)

“…Consequently, the allergenic venoms of fire ants of the genus Solenopsis (Myrmicinae), Pachycondyla spp. (Ponerinae), or Australian Myrmecia (Myrmeciinae) have especially been studied [5][6][7][8][9]. Also, the painful venoms of the bullet ant Paraponera clavata and of ponerine ants of the genera Dinoponera and Neoponera have been the subject of several studies which led to the isolation and characterization of tens of bioactive peptides [10,11].…”

Unravelling the potential of insects for medicinal purposes – A comprehensive review

“… Cicadidae periostracum - cyclic peptide [ 89 ] Hymenoptera Apidae Apis mellifera L. Venom - melittin [ 88 ] Apidae sp. Propolis - Caffeic acid phenethyl ester [ 90 ] Formicidae Polyrhachis dives Smith Dopamine derivatives [ 61 ] Myrmecia pilosula F. Smith Venom [ 171 ] Dinoponera quadriceps Kempf Venom [ [172] , [173] , [174] ] Scoliidae Scolia decorata ventralis Smith Venom - peptides [ 175 ] I00–I99 Blattodea Corydiidae Eupolyphaga sinensis Walker Serine proteases [ 176 ] Protein [ 176 ] Blattidae American cockroach L. Xinmailong [ 96 ] Hymenoptera Apidae Apis mellifera L. …”