Development of Dry Powder Formulations of AS01B containing vaccines using Thin-Film Freeze-Drying

AS01B is a liposomal formulation of two immunostimulants namely 3-O-desacyl-4’-monophosphoryl lipid A (MPL) and QS-21. The liposomal formulation of AS01B reduces the endotoxicity of MPL and the lytic activity of QS-21; however, it renders the adjuvant sensitive to accidental slow freezing. The liposomal formulation also represents a major challenge towards the formulation of dry powders of vaccines containing AS01B. In the present study, we tested the feasibility of applying thin-film freeze-drying (TFFD) to engineer dry powders of the AS01B liposomal adjuvant alone or vaccines containing AS01B as an adjuvant. Initially, we showed that after the AS01B liposomal adjuvant was subjected to TFFD using sucrose as a stabilizer at 4% w/v , the particle size distribution of AS01B liposomes reconstituted from the dry powder was identical to the liquid adjuvant before drying. We then showed using ovalbumin (OVA) as a model antigen adjuvanted with AS01B (AS01B/OVA) that subjecting the AS01B/OVA vaccine to TFFD and subsequent reconstitution did not negatively affect the AS01B liposome integrity, nor the immunogenicity of the vaccine. Importantly, the thin-film freeze-dried vaccine was not sensitive to repeated freezing-and-thawing. Finally, the feasibility of using TFFD to prepare dry powders of AS01B-adjuvanted vaccines was further confirmed using AS01B-adjuvanted Fluzone Quadrivalent and Shingrix, which contains AS01B. It is concluded that the TFFD technology can enable the formulation of AS01B-adjuvanted vaccines as freezing-insensitive dry powders in single-vial presentation. ### Competing Interest Statement ZC and ROW report financial support by TFF Pharmaceuticals, Inc. ZC reports a relationship with TFF Pharmaceuticals, Inc. that includes: equity or stocks and funding. ROW reports a relationship with TFF Pharmaceuticals, Inc. that includes: consulting or advisory, equity or stocks, and funding. HX report a relationship with TFF Pharmaceuticals, Inc. that includes: consulting or advisory. ZC and ROW have a patent (Dry solid aluminum adjuvant-containing vaccines and related methods thereof) pending to TFF Pharmaceuticals, Inc. ZC, ROW, KA and HX have a patent (Dry liposome adjuvant-containing vaccines and related methods thereof) pending to UT Austin. ZC, ROW and KA have a patent (Dry liposome formulations and related methods thereof) pending to UT Austin.