Periodic Reporting for period 3 - ENDFLU (Evaluation of ratioNally Designed Influenza vaccines)
Período documentado: 2023-08-01 hasta 2024-07-31
ENDFLU will develop three complementary next-generation influenza vaccination strategies for the world, fueled by the unique combined expertise of Indian and European partners in rational vaccine design and development:
• Create a repository of pre-pandemic GMP-compliant MVA-based vaccine seeds, each encoding an influenza hemagglutinin (HA), based on European partners demonstration of clinical safety and cross-clade immunogenicity of MVA-H5.
• Optimize and select protein- and MVA-based constructs developed by Indian and European ENDFLU partners, resulting in a rationally combined vaccine formulation that induces broadly protective humoral and cell-mediated immunity against all influenza manifestations. This will be advanced in a Phase I clinical trial and Controlled Human Infection Model (CHIM) study.
• Pre-clinically advance other promising protein- and MVA-based constructs toward future development beyond ENDFLU. Constructs to elicit humoral immunity build on rationally-designed and optimized conserved epitopes of influenza surface proteins. MVA-based constructs to elicit cell-mediated immunity will encode conserved internal protein T cell epitopes. The best candidates will be selected based on biochemical, biophysical, functional and manufacturing criteria, and immunogenicity and protective efficacy in mice and ferrets. The selected protein-based construct will enter a Part A Phase I clinical trial in India assessing safety, immunogenicity and optimal dose, to be combined with the selected MVA- based construct. This protein-MVA combination will enter a Part B Phase I clinical trial in the EU, assessing safety and immunogenicity, followed by the CHIM study. A dissemination and exploitation plan, and collaboration with key scientific advocacy organizations will facilitate the adoption of ENDFLU achievements to combat influenza worldwide.
• Create a repository of pre-pandemic GMP-compliant MVA-based vaccine seeds, each encoding an influenza hemagglutinin (HA), based on European partners demonstration of clinical safety and cross-clade immunogenicity of MVA-H5.
• Optimize and select protein- and MVA-based constructs developed by Indian and European ENDFLU partners, resulting in a rationally combined vaccine formulation that induces broadly protective humoral and cell-mediated immunity against all influenza manifestations. This will be advanced in a Phase I clinical trial and Controlled Human Infection Model (CHIM) study.
• Pre-clinically advance other promising protein- and MVA-based constructs toward future development beyond ENDFLU. Constructs to elicit humoral immunity build on rationally-designed and optimized conserved epitopes of influenza surface proteins. MVA-based constructs to elicit cell-mediated immunity will encode conserved internal protein T cell epitopes. The best candidates will be selected based on biochemical, biophysical, functional and manufacturing criteria, and immunogenicity and protective efficacy in mice and ferrets. The selected protein-based construct will enter a Part A Phase I clinical trial in India assessing safety, immunogenicity and optimal dose, to be combined with the selected MVA- based construct. This protein-MVA combination will enter a Part B Phase I clinical trial in the EU, assessing safety and immunogenicity, followed by the CHIM study. A dissemination and exploitation plan, and collaboration with key scientific advocacy organizations will facilitate the adoption of ENDFLU achievements to combat influenza worldwide.
H2, H5, H7, H9 and H10 synthetic target gene sequences have been designed, cloned into MVA vector plasmids, amplified under pre-GMP conditions and characterised in vitro, to establish an optimized repository of pre-GMP MVA-ENDFLU vector vaccines from influenza A viruses with pandemic potential.
Hemagglutination inhibition assay (HI), viral neutralization assay, ELISAs targeting HA stem/ectodomains and NA, and ELLA assays have been standardised, while ADCC assay is in progress. We have successfully expressed and purified recombinant ferret cytokines and developed a strategy for generating monoclonal antibodies against them, with strong potential for application beyond ENDFLU.
Protein-based constructs generated include HA (A/B) ectodomain fusions, fused with a disulfide-linked oligomerization domain or a folder oligomerization domain, HA stem domains updated to N22 WHO recommended sequences, M2e-NA fusion proteins, the generation of a scaffold for display of 4-fold symmetric proteins on ferritin, informed by deep-learning computational work, and chimeric hemagglutinin constructs.
MVA-based construct consists of a recombinant MVA expressing the full-length polyepitope (MVA-PE) with ubiquitin and with spacer, and was fully characterised in vitro. The encoding shuttle plasmid was transferred for pre-seed GLP batch preparation, and a patent was filed.
The selection of the final protein-based and MVA-based constructs for GMP production is made and consists of Mynflu001, Mynflu002 and MVA-PE.
GMP manufacturing of these constructs is in progress; the preparation of the Phase I clinical trial has been initiated.
Hemagglutination inhibition assay (HI), viral neutralization assay, ELISAs targeting HA stem/ectodomains and NA, and ELLA assays have been standardised, while ADCC assay is in progress. We have successfully expressed and purified recombinant ferret cytokines and developed a strategy for generating monoclonal antibodies against them, with strong potential for application beyond ENDFLU.
Protein-based constructs generated include HA (A/B) ectodomain fusions, fused with a disulfide-linked oligomerization domain or a folder oligomerization domain, HA stem domains updated to N22 WHO recommended sequences, M2e-NA fusion proteins, the generation of a scaffold for display of 4-fold symmetric proteins on ferritin, informed by deep-learning computational work, and chimeric hemagglutinin constructs.
MVA-based construct consists of a recombinant MVA expressing the full-length polyepitope (MVA-PE) with ubiquitin and with spacer, and was fully characterised in vitro. The encoding shuttle plasmid was transferred for pre-seed GLP batch preparation, and a patent was filed.
The selection of the final protein-based and MVA-based constructs for GMP production is made and consists of Mynflu001, Mynflu002 and MVA-PE.
GMP manufacturing of these constructs is in progress; the preparation of the Phase I clinical trial has been initiated.
ENDFLU's repository of (pandemic) seed vaccine candidates will be established based on the expression of influenza hemagglutinin (HA) by the Modified Vaccinia Ankara (MVA) vector. This is a technologically- and clinically advanced platform, with demonstrated broad intra-subtypic protective efficacy against influenza, elicited by humoral and cell-mediated immunity, both in animal models and in humans. The regulatory path towards their use upon the emergence of a new influenza pandemic will be established, in consultation with industry and regulatory authorities in EU (EMA) and India (CDSCO). Clonal isolation and amplification of recombinant MVA expressing H2, H5, H7, H9 and H10 under pre-GMP conditions is completed.
Pre-clinically well-advanced vaccine candidates will be optimized, by leveraging unique European and Indian scientific progress, strong expertise and know-how in the development of next-generation influenza vaccines. These include protein-based constructs based on a range of target influenza proteins and parts thereof as well as MVA constructs with conserved and/or cellular immunity targets. These ENDFLU vaccine candidates and other promising leads will be further pre-clinically advanced throughout the duration of the project. Those reaching rigorous and critical milestones for their eventual production under Good Laboratory Practice (GLP) conditions will be selected to enter GMP production and Phase I clinical studies. One protein-based vaccine candidate and one MVA-delivered vaccine candidate will be selected based on preclinical stability, safety, immunogenicity and protective efficacy data as well as criteria facilitating their rapid, economical, large-scale production and distribution worldwide, including in low- and middle-income countries (LMICs). The selection of the candidates for GMP production is ongoing.
The protein-based vaccine candidate will be tested for safety and immunogenicity in a Phase I dose escalation clinical trial in India. The dose that demonstrates the highest safety and immunogenicity profile will subsequently be combined with the MVA-delivered vaccine candidate as a single MVA-protein formulation to assess its safety and immunogenicity in a Phase I clinical trial, and subsequently to validate its protective efficacy in a CHIM study in the EU.
This approach takes advantage of promising influenza and unique vaccine development strategies of the Indian and European partners to ensure the synergistic development of a next generation rationally designed influenza vaccine candidate that will induce broad and potent humoral immunity against seasonal influenza A and B viruses through the protein-based component and robust and cross-reactive cell-mediated immunity through the MVA-based component.
Finally, a detailed exploitation plan, including a clinical development roadmap, will be prepared for continued clinical development of ENDFLU’s broadly protective influenza vaccine beyond the ENDFLU Phase I clinical trial.
Pre-clinically well-advanced vaccine candidates will be optimized, by leveraging unique European and Indian scientific progress, strong expertise and know-how in the development of next-generation influenza vaccines. These include protein-based constructs based on a range of target influenza proteins and parts thereof as well as MVA constructs with conserved and/or cellular immunity targets. These ENDFLU vaccine candidates and other promising leads will be further pre-clinically advanced throughout the duration of the project. Those reaching rigorous and critical milestones for their eventual production under Good Laboratory Practice (GLP) conditions will be selected to enter GMP production and Phase I clinical studies. One protein-based vaccine candidate and one MVA-delivered vaccine candidate will be selected based on preclinical stability, safety, immunogenicity and protective efficacy data as well as criteria facilitating their rapid, economical, large-scale production and distribution worldwide, including in low- and middle-income countries (LMICs). The selection of the candidates for GMP production is ongoing.
The protein-based vaccine candidate will be tested for safety and immunogenicity in a Phase I dose escalation clinical trial in India. The dose that demonstrates the highest safety and immunogenicity profile will subsequently be combined with the MVA-delivered vaccine candidate as a single MVA-protein formulation to assess its safety and immunogenicity in a Phase I clinical trial, and subsequently to validate its protective efficacy in a CHIM study in the EU.
This approach takes advantage of promising influenza and unique vaccine development strategies of the Indian and European partners to ensure the synergistic development of a next generation rationally designed influenza vaccine candidate that will induce broad and potent humoral immunity against seasonal influenza A and B viruses through the protein-based component and robust and cross-reactive cell-mediated immunity through the MVA-based component.
Finally, a detailed exploitation plan, including a clinical development roadmap, will be prepared for continued clinical development of ENDFLU’s broadly protective influenza vaccine beyond the ENDFLU Phase I clinical trial.