Prefusion RSV F proteins and their use
Inventors
Kwong, Peter • Graham, Barney • Mascola, John • Ou, Li • Druz, Aliaksandr • Chen, Man • Kong, Wing-pui • Georgiev, Ivelin Stefanov • Rundlet, Emily • Joyce, Michael Gordon • Tsybovsky, Yaroslav • Thomas, Paul • Pancera, Marie • Sastry, Mallika • Soto, Cinque • Van Galen, Joseph • Stewart-Jones, Guillaume • Yang, YongPing • Zhang, Baoshan • Baxa, Ulrich
Assignees
US Department of Health and Human Services
Publication Number
US-12275758-B2
Publication Date
2025-04-15
Expiration Date
2037-03-29
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
Embodiments of a recombinant Respiratory Syncytial Virus (RSV) F ectodomain trimer stabilized in a prefusion conformation are provided. Also disclosed are nucleic acids encoding the RSV F ectodomain trimer and methods of producing the RSV F ectodomain trimer. Methods for inducing an immune response in a subject are also disclosed. In some embodiments, the method can be a method for treating or preventing a RSV infection in a subject by administering a therapeutically effective amount of the recombinant RSV F ectodomain trimer to the subject.
Core Innovation
Embodiments of recombinant Respiratory Syncytial Virus (RSV) F ectodomain trimers stabilized in a prefusion conformation are provided, including nucleic acids encoding these trimers and methods of their production. These recombinant RSV F ectodomain trimers are useful for inducing an immune response in subjects, particularly for preventing or reducing RSV infection through administration of a therapeutically effective amount of the recombinant protein.
The problem addressed relates to RSV being a major cause of bronchiolitis and pneumonia in infants, as well as causing repeated infections across ages, especially in the elderly and immunocompromised. While the RSV F protein in prefusion conformation induces a greater neutralizing immune response than post-fusion conformations, existing immunogens such as DS-Cav1, which stabilize the prefusion conformation with certain substitutions, still leave room for improved immunogenicity and stability to better prevent severe disease and support maternal immunization protocols.
The invention discloses recombinant RSV F proteins developed via iterative structure-based design methods that further increase RSV-protective titers approximately fourfold beyond prior immunogens like DS-Cav1. These novel immunogens include genetically linked F1 and F2 subunits with the fusion peptide deleted, specific cavity-filling and disulfide bond substitutions to stabilize the trimeric protein in the prefusion state. Additionally, these recombinant proteins exhibit superior manufacturing attributes such as no requirement for furin cleavage and enhanced antigenic stability at high temperatures, with certain embodiments showing over tenfold improved stability compared to DS-Cav1 at 60° C. This enhanced combination of immunogenicity and stability is suitable for applications like maternal immunization to provide passive immunity to newborn infants.
Claims Coverage
The claims disclose multiple inventive features primarily related to RNA and mRNA molecules encoding recombinant RSV F proteins with specific stabilizing mutations, as well as immunogenic compositions and methods of inducing immune responses. Key inventive features involve the combination of deletions, peptide linkers, amino acid substitutions, and inter-protomer disulfide bonds for stabilization in the prefusion conformation.
RNA molecule encoding stabilized recombinant RSV F protein with specific deletions, linkers, and amino acid substitutions
An RNA molecule encoding a recombinant RSV F protein with deletion of positions 104-144 and a glycine-serine peptide linker between positions 103 and 145, and specific amino acid substitutions S155C, S290C, S190F, V207L, A149C, Y458C, corresponding to a reference sequence (SEQ ID NO: 57), with the recombinant protein comprising specified residue ranges of sequences set forth as SEQ ID NOs: 1, 5, 10, 15, or 20.
Inclusion of transmembrane domain and cytoplasmic tail in encoded RSV F protein
The RNA molecule encodes an RSV F protein that includes an RSV F transmembrane domain and cytoplasmic tail, or a precursor protein comprising a signal peptide, F2 polypeptide, F1 ectodomain, and transmembrane domain and cytosolic tail.
Immunogenic compositions comprising the RNA molecules and methods of inducing immune response
Immunogenic compositions including the RNA molecules encoding stabilized RSV F proteins, and methods of inducing an immune response to RSV F protein in subjects by administering an effective amount of such RNA molecules to generate the immune response.
Messenger RNA molecule encoding stabilized recombinant RSV F protein with similar deletions, linkers, and amino acid substitutions
A messenger RNA molecule encoding a recombinant RSV F protein with the same deletion and peptide linker between positions 103 and 145, amino acid substitutions S155C, S290C, S190F, V207L, A149C, Y458C, according to reference sequence SEQ ID NO: 57, comprising amino acid sequences as residues 26-474 or 31-479 of SEQ ID NOs: 1, 5, 10, 15, or 20.
Immunogenic compositions comprising the mRNA molecules and methods of inducing immune response
Immunogenic compositions including the mRNA molecules encoding stabilized RSV F proteins, and methods of inducing an immune response in a subject by administering an effective amount of such mRNA molecules.
RNA molecule encoding recombinant RSV F protein with alternative inter-protomer disulfide substitutions in absence of certain cysteine substitutions
An RNA molecule encoding a recombinant RSV F protein with deletion of positions 104-144 and a glycine-serine peptide linker, amino acid substitutions S155C, S290C, S190F, V207L, and either (A) Q98C and Q361C substitutions, or (B) N183GC and N428C substitutions, according to reference sequence SEQ ID NO: 57, with specified residue ranges from other SEQ IDs set forth in the claims.
The claims cover RNA and mRNA molecules encoding recombinant RSV F proteins with specific engineered deletions, peptide linkers, and amino acid substitutions including inter-protomer disulfide bonds to stabilize the protein in a prefusion conformation, as well as related immunogenic compositions and methods of inducing immune responses in subjects, including humans and bovines, for prevention or treatment of RSV infection.
Stated Advantages
Increased RSV-protective titers approximately fourfold higher than prior RSV F-based immunogens such as DS-Cav1.
Absence of a requirement for furin cleavage for the recombinant RSV F protein.
Increased antigenic stability to heat inactivation with several embodiments over tenfold more stable than DS-Cav1 at 60° C.
An unexpectedly superior combination of immunogenicity and stability.
Documented Applications
Use of recombinant RSV F ectodomain trimers as vaccine antigens to induce an immune response to RSV.
Administration to a subject for treating or preventing an RSV infection.
Maternal immunization protocols to induce immune response in pregnant subjects that provide passive immunity to infants against RSV infection in the first six months of life.
Use in prime-boost vaccination protocols, including DNA-primer and protein-boost vaccination approaches.
Use of recombinant RSV F ectodomain trimers on protein nanoparticles or virus-like particles for immunization.
Interested in licensing this patent?