Modified RSV F proteins and methods of their use
Inventors
Pushko, Peter M. • Wu, Yingyun • MASSARE, Michael J. • Liu, Ye • Smith, Gale • Zhou, Bin
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11446374-B2
Publication Date
2022-09-20
Expiration Date
Abstract
The present invention is generally related to modified or mutated respiratory syncytial virus fusion (F) proteins and methods for making and using them, including immunogenic compositions such as vaccines for the treatment and/or prevention of RSV infection.
Core Innovation
The invention relates to a modified RSV fusion (F) glycoprotein used in a vaccine composition and/or immunogenic composition. The modified RSV F glycoprotein includes an inactivated primary furin cleavage site and a wild-type secondary furin cleavage site having an amino acid sequence of RARR (SEQ ID NO: 23). The primary cleavage site is positioned at amino acids 131 to 136, and the secondary cleavage site is positioned at amino acids 106 to 109.
The primary furin cleavage site is inactivated by amino acid substitution of one or more amino acids of arginine 133, arginine 135 and arginine 136. In the described recombinant variants, the RSV F glycoprotein is cleaved into F1 and F2 subunits which are disulfide linked, and a p27 protein is connected to the F1 subunit via an amide bond. The RSV F glycoprotein lacks an N-terminal signal peptide.
The invention further includes variants with a partial deletion in the F1 fusion domain, deleting residues 137 to 146. The document describes expression and formulation of recombinant RSV F spike structures, including purified lollipop-form F spikes in rosette micelles and formulations including virus-like particles (VLPs). Additional product formats include isolated nucleic acids, vectors, cells, and baculovirus vectors.
Claims Coverage
The independent claims identified from the provided material are directed to a vaccine composition comprising a specifically engineered RSV F glycoprotein and an adjuvant. The coverage centers on an inactivated primary furin cleavage site with a wild-type secondary furin cleavage site, defined cleavage into disulfide-linked F1 and F2 subunits with a p27 connection via an amide bond, and additional structural constraints that further narrow the RSV F glycoprotein features.
Vaccine composition with engineered RSV F furin cleavage sites and defined cleavage linkage
A vaccine composition comprising an RSV F glycoprotein having an inactivated primary furin cleavage site and a wild-type secondary furin cleavage site having an amino acid sequence of RARR (SEQ ID NO: 23), wherein the primary cleavage site is at amino acids 131 to 136 and the secondary cleavage site is at amino acids 106 to 109, and wherein the RSV F glycoprotein lacks an N-terminal signal peptide, is cleaved into disulfide linked F1 and F2 subunits, and includes a p27 protein connected to the F1 subunit via an amide bond.
Only the RSV F glycoprotein as the viral glycoprotein in the composition
The vaccine composition wherein the only viral glycoprotein in the composition is the RSV F glycoprotein, together with an adjuvant.
Inactivation of the primary furin cleavage site by arginine substitution at positions 133, 135, and 136
The RSV F glycoprotein wherein the primary furin cleavage site is inactivated by amino acid substitution of one or more amino acids of arginine 133, arginine 135 and arginine 136.
Alum as the adjuvant
The vaccine composition wherein the adjuvant is alum.
Specific primary furin cleavage site substitutions with glutamine at arginine positions 133, 135, and 136
The vaccine composition wherein amino acids arginine 133, arginine 135 and arginine 136 are substituted with glutamine.
Limited deletion in the RSV F1 fusion domain corresponding to residues 137 to 146
The vaccine composition wherein the RSV F glycoprotein includes a deletion of up to 10 amino acids that corresponds to the fusion domain spanning residues 137 to 146 of SEQ ID NO: 2.
RSV F glycoprotein includes a transmembrane domain
The vaccine composition characterized by an RSV F glycoprotein that includes a transmembrane domain.
Across the provided claim coverage, the principal inventive content is a vaccine composition defined by a specifically engineered RSV F glycoprotein with an inactivated primary furin cleavage site and a wild-type secondary furin cleavage site (RARR), defined cleavage into disulfide-linked F1 and F2 subunits with p27 connected to F1 via an amide bond, and constraints on viral glycoprotein content and signal peptide presence, with dependent refinements that specify the adjuvant (alum), define particular arginine-to-glutamine substitutions, and further constrain RSV F structure through fusion-domain deletion and/or inclusion of a transmembrane domain.
Stated Advantages
Enhanced RSV neutralization with alum adjuvant.
Lung RSV inhibition after challenge in mice.
Robust neutralizing responses in cotton rats for RSV-A and RSV-B.
Reduced inflammation versus FI-RSV.
Improved expression and purity.
Micelle morphology and size characterization consistent with the described formulations.
Documented Applications
Vaccine and/or immunogenic compositions comprising the modified RSV F glycoprotein and an adjuvant.
Use in challenge settings including lung RSV inhibition after challenge in mice.
Use in neutralization/challenge contexts including cotton rats challenged with RSV-A and RSV-B.
Formulation of virus-like particles (VLPs) including RSV F together with BRSV M and/or HRSV N.
Interested in licensing this patent?