Rational design of heat stable lyophilized rotavirus vaccine formulations

Abstract

To develop a safe and efficacious heat-stable rotavirus vaccine, new lyophilized formulations were developed using rotavirus serotypes constituting RotaTeq®. A series of formulation compositions, differing in buffering agents, bulking agents, cryoprotectants, amino acids and divalent cations, were screened for their ability to provide stability to rotavirus serotypes during lyophilization and when stored under elevated temperatures for extended periods. Lead formulations and lyophilization cycles were further optimized. Stability profiles of thus optimized formulations showed their ability to retain the potency of rotavirus for > 36 months at 5°C, 20 months at 37°C, and 7 months at 45°C. The heat-stable lyophilized rotavirus formulations developed met the all critical quality attributes for appearance, heat-stability during storage, moisture content as well as pH, viability and stability after reconstitution and has great potential to be used as vaccine candidates for improving access in low-income countries.

Thermostabilization of viruses via complex coacervation

“…However, these methods typically require the presence of sugars (e.g., sucrose, mannitol, and trehalose), amino acids, and/or other cryoprotectants and bulking agents that tend to hydrogen bond with viral capsid proteins and/or viral envelopes to entrain surface-bound water and form a stabilizing matrix. [15][16][17][18][19] For example, lyophilized rotavirus vaccines formulated in optimized buffer conditions with polyvinyl pyrrolidone as a bulking agent, sucrose as a cryoprotectant, and L-arginine and glycine as osmolytes, can retain potency for 20 months at 37°C and 7 months at 45°C. 18 An analogous strategy where viral encapsulation in hydrated silica was used in place of an organic matrix slowed the infectivity loss of the human enterovirus type 71 by six-fold at 37°C for 20 days, or at 40°C for 36 hours.…”

“…[15][16][17][18][19] For example, lyophilized rotavirus vaccines formulated in optimized buffer conditions with polyvinyl pyrrolidone as a bulking agent, sucrose as a cryoprotectant, and L-arginine and glycine as osmolytes, can retain potency for 20 months at 37°C and 7 months at 45°C. 18 An analogous strategy where viral encapsulation in hydrated silica was used in place of an organic matrix slowed the infectivity loss of the human enterovirus type 71 by six-fold at 37°C for 20 days, or at 40°C for 36 hours. 20 Although such formulation methods show promise for thermostabilizing vaccines, the outcomes tend to be the result of large-scale trial and error experiments, and there is a need for a simple, low-cost, and versatile approach for stabilizing viruses.…”

A randomized Phase I/II study to evaluate safety and reactogenicity of a heat-stable rotavirus vaccine in healthy adults followed by evaluation of the safety, reactogenicity, and immunogenicity in infants

“…The HSRV formulation offers a stability profile of 7 months at 45°C (first of its kind stability profile reported anywhere for any rotavirus vaccine) 14 and 20 months at 37°C…”

“…HSRV was formulated as a lyophilized cake comprising of GRAS (generally regarded as safe) excipients as described previously. 14 A series of formulation compositions, differing in buffering agents, bulking agents, cryoprotectants, amino acids, and divalent cations, were screened for their ability to provide stability to rotavirus serotypes during lyophilization and when stored under-elevated temperatures for extended periods before arriving at the final lead formulation 14 HSRVused in this study. This lyophilized cake was reconstituted using a reconstitution buffer just prior to administration.…”

“…Lyophilized HSRV vaccine vials from the same batch were kept in stability chambers (Memmert, Germany) at various temperature conditions of 2-8°C, 25°C, 37°C, and 45°C for more than 36 months and assessed periodically for stability by multivalent qPCR-based rotavirus potency assay (M-QPA) as described previously. 14 The composition concentration per dose (2 mL) of the reconstituted HSRV is listed in Table 2. Each of the five constituent rotavirus serotypes of HSRV were viable upon reconstitution as determined by M-QPA assay.…”