New GMP manufacturing processes to obtain thermostable HIV-1 gp41 virosomes under solid forms for various mucosal vaccination routes

Mario Amacker,Mark Spengler,Markus Schwaller,Richard Johnson,Laura Mason,Toon Stegmann,Michael Adler,Charli Smardon,Beena Punnamoottil,Kirk Jeffery,Farien Bhoelan,Olivia Bonduelle,Jack Sorrell,Andrew Naylor,Behazine Combadière,Desmond Wong,Sylvain Fleury,Olga Belova

doi:10.1038/s41541-020-0190-9

Abstract

The main objective of the MACIVIVA European consortium was to develop new Good Manufacturing Practice pilot lines for manufacturing thermostable vaccines with stabilized antigens on influenza virosomes as enveloped virus-like particles. The HIV-1 gp41-derived antigens anchored in the virosome membrane, along with the adjuvant 3M-052 (TLR7/8 agonist) on the same particle, served as a candidate vaccine for the proof of concept for establishing manufacturing processes, which can be directly applied or adapted to other virosomal vaccines or lipid-based particles. Heat spray-dried powders suitable for nasal or oral delivery, and freeze-dried sublingual tablets were successfully developed as solid dosage forms for mucosal vaccination. The antigenic properties of vaccinal antigens with key gp41 epitopes were maintained, preserving the original immunogenicity of the starting liquid form, and also when solid forms were exposed to high temperature (40 °C) for up to 3 months, with minimal antigen and adjuvant content variation. Virosomes reconstituted from the powder forms remained as free particles with similar size, virosome uptake by antigen-presenting cells in vitro was comparable to virosomes from the liquid form, and the presence of excipients specific to each solid form did not prevent virosome transport to the draining lymph nodes of immunized mice. Virosome integrity was also preserved during exposure to <−15 °C, mimicking accidental freezing conditions. These “ready to use and all-in-one” thermostable needle-free virosomal HIV-1 mucosal vaccines offer the advantage of simplified logistics with a lower dependence on the cold chain during shipments and distribution.

Highlights

The majority of the world population lives in warm regions, and for low and middle-income countries, maintaining the cold chain for preserving biological products is challenging due to unreliable electricity access and inadequate or limited storage facilities
Because they have a similar size and shape to viruses, they belong to the enveloped virus-like particles family
The new adjuvanted human immunodeficiency virus type 1 (HIV-1) vaccine called MYM-V202 contains two types of virosomes, one that displays P1 and the other one with rgp[41], with the adjuvant anchored on the same virosome particle to minimize non-specific immune activation and further improves the vaccine tolerance and safety

Summary

INTRODUCTION

The majority of the world population lives in warm regions, and for low and middle-income countries, maintaining the cold chain for preserving biological products is challenging due to unreliable electricity access and inadequate or limited storage facilities. Because HIV-1 replicates in various mucosal tissues, an HIV subunit vaccine allowing a prime/boost approach, combining two distinct mucosal sites, could more efficiently achieve a broader mucosal tissue coverage in both men and women This explains the strong interest in developing various new galenic virosomal formulations under thermostable solid dosages for mucosal delivery, as early studies with liquid nasal[33,57] and sublingual (SL)[26,58] virosomes induced systemic and mucosal antibodies. The new adjuvanted HIV-1 vaccine called MYM-V202 contains two types of virosomes, one that displays P1 and the other one with rgp[41], with the adjuvant anchored on the same virosome particle to minimize non-specific immune activation and further improves the vaccine tolerance and safety With this new galenic formulation, we have verified that the new excipients were not detrimental to virosome particles, once delivered in vivo for the vaccine-induced immune responses. Both IM effect because they have been evaluated extensively in diverse and ID routes were attractive and tested for monitoring the studies, certain excipients such as the trehalose may represent virosome migration to the draining lymph nodes (Fig. 3)

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DISCUSSION

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METHODS