CA-3291895-A1 - PREFUSION RSV F PROTEINS AND THEIR USE

Abstract

Disclosed are Respiratory Syncytial Virus (RSV) antigens including a recombinant RSV F protein stabilized in a prefusion conformation. Also disclosed are nucleic acids encoding the antigens and methods of producing the antigens. Methods for generating an immune response in a subject are also disclosed. In some embodiments, the method is a method for treating or preventing a RSV infection in a subject by administering a therapeutically effective amount of the antigen to the subject.

Inventors

• Peter D. Kwong
• Cinque Soto
• Sanjay Srivatsan
• Guillaume Stewart-Jones
• Lei Chen
• Man Chen
• Gwo-Yu Chuang
• Jason Gorman
• Gilad Ofek
• Mallika Sastry
• YongPing Yang
• Barney S. Graham
• Tongqing Zhou
• Jason S. McLellan
• Michael Gordon Joyce
• Masaru Kanekiyo
• Baoshan Zhang
• Jeffrey Boyington
• Ivelin Stefanov Georgiev
• Marie Pancera

Assignees

• THE UNITED STATES OF AMERICA, AS REPRESENTED BY THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES

Dates

Publication Date

20260302

Application Date

20140313

Priority Date

20130313

Claims (1)

1. <pat:ClaimStatement>CLAIMS:</pat:ClaimStatement> <pat:Claims com:id="claims"> <pat:Claim com:id="CLM-00001"> <pat:ClaimNumber>1</pat:ClaimNumber> <pat:ClaimText>1. An isolated immunogen, comprising a recombinant RSV F protein or an extracellular domain thereof comprising a S190I substitution that stabilizes the recombinant RSV F protein or an extracellular domain thereof in a prefusion conformation, wherein the amino acid positions of the RSV F protein are according to a reference RSV F protein sequence set forth as SEQ ID NO: 124. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00002"> <pat:ClaimNumber>2</pat:ClaimNumber> <pat:ClaimText>2. The immunogen of claim 1, wherein the recombinant RSV F protein or extracellular domain thereof comprises an F2 polypeptide and an F1 polypeptide comprising amino acid sequences at least 90% identity to residues 26-109 and 137-513, respectively, or 26- 103 and 145-513, respectively, of SEQ ID NO: 910. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00003"> <pat:ClaimNumber>3</pat:ClaimNumber> <pat:ClaimText>3. The immunogen of claim 1 or claim 2, wherein the immunogen specifically binds to a D25 or a AM22 prefusion-specific antibody. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00004"> <pat:ClaimNumber>4</pat:ClaimNumber> <pat:ClaimText>4. The immunogen of any one of claims 1-3, wherein the RSV F protein or extracellular domain thereof is a RSV A, B, or bovine RSV F protein or extracellular domain thereof comprising the amino acid substitution. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00005"> <pat:ClaimNumber>5</pat:ClaimNumber> <pat:ClaimText>5. The immunogen of any one of claims 1-4, wherein the recombinant RSV F protein or extracellular domain thereof is a single chain protein comprising F₂ and F₁ polypeptides linked by a heterologous peptide linker, or directly linked. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00006"> <pat:ClaimNumber>6</pat:ClaimNumber> <pat:ClaimText>6. The immunogen of claim 5, wherein position 105 of the F₂ polypeptide is linked to position 145 of the F₁ polypeptide by a Gly-Ser linker; or position 103 of the F₂ polypeptide is directly linked to position 145 of the F₁ polypeptide. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00007"> <pat:ClaimNumber>7</pat:ClaimNumber> <pat:ClaimText>7. The immunogen of any one of claims 1-6, comprising a multimer of the recombinant RSV F protein or extracellular domain thereof. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00008"> <pat:ClaimNumber>8</pat:ClaimNumber> <pat:ClaimText>8. The immunogen of any one of claims 1-7, wherein the recombinant RSV F protein or extracellular domain thereof is linked to a trimerization domain. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00009"> <pat:ClaimNumber>9</pat:ClaimNumber> <pat:ClaimText>9. The immunogen of claim 8, wherein the trimerization domain is a Foldon domain. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00010"> <pat:ClaimNumber>10</pat:ClaimNumber> <pat:ClaimText>10. The immunogen of claim 1, wherein a C-terminal residue of an F₁ polypeptide of the recombinant RSV F protein or extracellular domain thereof is linked to a foldon trimerization domain. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00011"> <pat:ClaimNumber>11</pat:ClaimNumber> <pat:ClaimText>11. The immunogen of claim 8, comprising the extracellular domain of the RSV F protein linked to the trimerization domain, and an amino acid sequence at least 90% identical to residues 26-109 and 137-544 or 26-103 and 145-544 of SEQ ID NO: 910. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00012"> <pat:ClaimNumber>12</pat:ClaimNumber> <pat:ClaimText>12. The immunogen of any one of claims 1-7, wherein a C-terminal residue of an F₁ polypeptide of the recombinant RSV F protein or extracellular domain thereof is linked to a transmembrane domain. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00013"> <pat:ClaimNumber>13</pat:ClaimNumber> <pat:ClaimText>13. The immunogen of any one of claims 1-12, wherein a Fab of monoclonal antibody D25 or AM22 specifically binds to the immunogen with a K_d of 1 µM or less. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00014"> <pat:ClaimNumber>14</pat:ClaimNumber> <pat:ClaimText>14. A virus-like particle comprising the immunogen of any one of claims 1-13. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00015"> <pat:ClaimNumber>15</pat:ClaimNumber> <pat:ClaimText>15. A protein nanoparticle comprising the immunogen of any one of claims 1-13. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00016"> <pat:ClaimNumber>16</pat:ClaimNumber> <pat:ClaimText>16. The protein nanoparticle of claim 15, wherein the protein nanoparticle is a ferritin nanoparticle, an encapsulin nanoparticle, a Sulfur Oxygenase Reductase (SOR) nanoparticle, a lumazine synthase nanoparticle or a pyruvate dehydrogenase nanoparticle. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00017"> <pat:ClaimNumber>17</pat:ClaimNumber> <pat:ClaimText>17. A nucleic acid molecule encoding the immunogen of any one of claims 1-13. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00018"> <pat:ClaimNumber>18</pat:ClaimNumber> <pat:ClaimText>18. The nucleic acid molecule of claim 17, wherein the nucleic acid molecule encodes a precursor protein of the immunogen or protein nanoparticle. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00019"> <pat:ClaimNumber>19</pat:ClaimNumber> <pat:ClaimText>19. The nucleic acid molecule of claim 17 or claim 18, wherein the nucleic acid molecule is an RNA molecule. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00020"> <pat:ClaimNumber>20</pat:ClaimNumber> <pat:ClaimText>20. A vector comprising the nucleic acid molecule of any one of claims 17-19. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00021"> <pat:ClaimNumber>21</pat:ClaimNumber> <pat:ClaimText>21. The vector of claim 20, wherein the vector is a viral vector. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00022"> <pat:ClaimNumber>22</pat:ClaimNumber> <pat:ClaimText>22. An isolated host cell comprising the vector of claim 20. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00023"> <pat:ClaimNumber>23</pat:ClaimNumber> <pat:ClaimText>23. An immunogenic composition comprising an effective amount of the immunogen of any one of claims 1-13, the virus-like particle of claim 14, the protein nanoparticle of claim 15 or claim 16, the nucleic acid molecule of any one of claims 17-19, or the vector of claim 20 or claim 21, and a pharmaceutically acceptable carrier. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00024"> <pat:ClaimNumber>24</pat:ClaimNumber> <pat:ClaimText>24. The immunogen of any one of claims 1-13 for use in generating an immune response to RSV F protein in a subject. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00025"> <pat:ClaimNumber>25</pat:ClaimNumber> <pat:ClaimText>25. An isolated immunogen, comprising: an extracellular domain of an RSV F protein fused to a foldon trimerization domain, wherein: the extracellular domain of the RSV F protein comprises a S190I substitution; the extracellular domain of the RSV F protein fused to the foldon trimerization domain comprises an amino acid sequence at least 90% identical to residues 26-109 and 137-513, respectively, or 26-103 and 145-513, respectively, of SEQ ID NO: 910; and wherein the amino acid positions are according to a reference RSV F protein sequence set forth as SEQ ID NO: 124. </pat:ClaimText> </pat:Claim> </pat:Claims>