Prefusion coronavirus spike proteins and their use
Inventors
Graham, Barney • McLellan, Jason • Ward, Andrew • Kirchdoerfer, Robert • Cottrell, Christopher • Joyce, Michael Gordon • Kanekiyo, Masaru • Wang, Nianshuang • Pallesen, Jesper • Yassine, Hadi • Turner, Hannah • Corbett, Kizzmekia
Assignees
Dartmouth College • Scripps Research Institute • US Department of Health and Human Services
Publication Number
US-11964010-B2
Publication Date
2024-04-23
Expiration Date
2037-10-25
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
Coronavirus S ectodomain trimers stabilized in a prefusion conformation, nucleic acid molecules and vectors encoding these proteins, and methods of their use and production are disclosed. In several embodiments, the coronavirus S ectodomain trimers and/or nucleic acid molecules can be used to generate an immune response to coronavirus in a subject. In additional embodiments, the therapeutically effective amount of the coronavirus S ectodomain trimers and/or nucleic acid molecules can be administered to a subject in a method of treating or preventing coronavirus infection.
Core Innovation
The invention relates to recombinant coronavirus spike (S) proteins, particularly alphacoronavirus or betacoronavirus S ectodomain trimers, that are stabilized in the prefusion conformation by one or more amino acid substitutions, especially proline substitutions. These S ectodomain trimers comprise protomers each containing one or two consecutive proline substitutions at or near the junction between a heptad repeat 1 (HR1) domain and a central helix domain, which stabilize the S protein trimer in the prefusion conformation. This structural stabilization prevents the conformational change from prefusion to postfusion state, thereby maintaining a more immunogenic and stable structure. Nucleic acid molecules and vectors encoding these stabilized proteins, as well as methods for their production and use, are also disclosed.
The problem addressed arises from the high pathogenicity and airborne transmissibility of coronaviruses such as MERS-CoV and SARS-CoV, which have caused outbreaks with high case-fatality rates and pose a potential threat for future pandemics. Prior to this invention, effective vaccines and therapeutic agents against these coronaviruses were lacking. The instability of the coronavirus S protein, which undergoes conformational changes between prefusion and postfusion states, complicates vaccine design. The invention specifically solves this by engineering recombinant S ectodomain trimers stabilized in the prefusion conformation, which are more effective immunogens, producing superior immune responses as demonstrated in animal models compared to unstabilized native proteins.
Additional features involve specific mutations, such as double proline substitutions at residues corresponding to those near or within HR1 and the central helix region, tailored to different coronavirus strains including MERS-CoV (e.g., V1060P and L1061P), SARS-CoV (e.g., K968P and V969P), HKU1-CoV, OC43-CoV, and others. These mutations increase protein expression, stability, and induce stronger neutralizing antibody responses. The stabilized S ectodomain trimers can be produced as soluble proteins, linked to trimerization domains like the T4 fibritin foldon, linked to transmembrane domains for membrane anchoring, or displayed on protein nanoparticles to enhance immunogenicity.
Claims Coverage
The independent claims focus on nucleic acid molecules encoding recombinant coronavirus S proteins with specific stabilizing proline substitutions, and their use in immunogenic compositions and methods of generating immune responses.
Nucleic acid encoding recombinant coronavirus S protein with double proline substitutions
A nucleic acid molecule encoding a recombinant coronavirus S protein comprising two proline substitutions located between the heptad repeat 1 (HR1) and the central helix, stabilizing the protein in the prefusion conformation.
Betacoronavirus and broad coronavirus strain applicability
The nucleic acid molecule encodes S proteins from betacoronaviruses including MERS-CoV, SARS-CoV, and several other related coronaviruses, encompassing multiple species and strains.
Additional mutation to inhibit S1/S2 cleavage
The recombinant coronavirus S protein further comprises mutations that inhibit cleavage at the S1/S2 protease cleavage site, enhancing stability of the prefusion conformation.
Immunogenic compositions including the nucleic acid molecules
Formulations comprising the nucleic acid molecule and a pharmaceutically acceptable carrier, enabling administration as vaccines or immunogens.
Methods of generating immune responses
Methods of administering an effective amount of the nucleic acid molecule to a subject to induce an immune response to the coronavirus S protein, which can inhibit coronavirus infection.
mRNA molecules encoding stabilized S proteins
Nucleic acid molecules can be mRNA encoding the stabilized coronavirus S proteins with the double proline substitutions and optional cleavage site mutations.
The claims comprehensively protect nucleic acid molecules encoding recombinant coronavirus S proteins stabilized by double proline substitutions near the HR1-central helix interface, methods for immunizing subjects with these molecules, immunogenic formulations, and widen applicability to multiple coronaviruses and nucleic acid types including mRNA.
Stated Advantages
The prefusion-stabilized coronavirus S ectodomain trimers produce a superior immune response in animal models compared to corresponding non-stabilized trimers.
The amino acid substitutions, particularly double proline substitutions, increase expression yields and thermal stability of the recombinant S proteins.
The stabilized S ectodomain trimers elicit broader and higher potency neutralizing antibody responses targeting multiple epitopes.
Immunization with the prefusion stabilized MERS-CoV S ectodomain trimer protects animals from lethal MERS-CoV challenge.
Documented Applications
Use of the recombinant coronavirus S ectodomain trimers or nucleic acid molecules encoding them to induce an immune response in a subject against the corresponding coronavirus.
Use in immunogenic compositions and vaccines for treating, preventing, or inhibiting coronavirus infection including MERS-CoV and SARS-CoV.
Use of S ectodomain trimers linked to trimerization domains or displayed on protein nanoparticles as vaccine antigens.
Incorporation into viral vectors or virus-like particles for immunization strategies.
Interested in licensing this patent?