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CN-122029184-A - Agents and methods for preventing, treating or limiting henipav infection

CN122029184ACN 122029184 ACN122029184 ACN 122029184ACN-122029184-A

Abstract

Isolated polypeptides and nucleic acids encoding them are provided, wherein the polypeptides comprise (a) an attached glycoprotein G Head Domain (GHD) comprising an amino acid sequence at least 70% identical to the amino acid sequence of any of SEQ ID NOs 1-2, and (b) a multimerization domain capable of producing multimers comprising 2 to 60 copies of the isolated polypeptides, and methods of using the polypeptides and nucleic acids to treat or limit the progression of a henapa virus infection.

Inventors

  • P. Brandis

Assignees

  • 菲莱克斯生物科学有限公司

Dates

Publication Date
20260512
Application Date
20240917
Priority Date
20230919

Claims (20)

  1. 1. An isolated polypeptide comprising: (a) An attached glycoprotein G Head Domain (GHD) comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of any of SEQ ID NO:1, optionally wherein the head domain comprises the amino acid sequence of SEQ ID NO:1, wherein at least 1, 2, 3,4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or all 20 residues of the residues defined by brackets are the first listed residue, and (B) A multimerization domain capable of producing a multimer comprising 2 to 60 copies of the isolated polypeptide.
  2. 2. The isolated polypeptide of claim 1, wherein the multimerization domain produces 4 copies or 60 copies of the isolated polypeptide.
  3. 3. The isolated polypeptide of claim 1 or 2, wherein the multimerization domain comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO 3-4, optionally wherein the multimerization structure comprises the amino acid sequence of SEQ ID NO 3, wherein at least 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or all 18 residues of the residues defined by brackets are the first listed residues.
  4. 4. The isolated polypeptide of any one of claims 1-3, further comprising an amino acid linker located between the head domain and the multimerization domain.
  5. 5. The isolated polypeptide of claim 4, wherein the linker comprises or consists of (GGS) n GGG, wherein n is 3-11、4-11、5-11、6-11、7-11、8-11、9-11、10-11、3-10、4-10、5-10、6-10、7-10、8-10、9-10、3-9、4-9、5-9、6-9、7-9、8-9、3-8、4-8、5-8、6-8、7-8、3、4、5、6、7、8、9、10 or 11, optionally wherein n is 8-11, 9-11, 10-11, 8-10, 9-10, 8-9, 8, 9, 10 or 11.
  6. 6. The isolated polypeptide of any one of claims 1-5, wherein the head domain is located at the carboxy terminus of the multimerization domain.
  7. 7. The isolated polypeptide of any of claims 1-6, comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs 5-6, wherein n is 3-11、4-11、5-11、6-11、7-11、8-11、9-11、10-11、3-10、4-10、5-10、6-10、7-10、8-10、9-10、3-9、4-9、5-9、6-9、7-9、8-9、3-8、4-8 6-8、7-8、3、4、5、6、7、8、9、10 or 11, optionally wherein n is 8-11, 9-11, 10-11, 8-10, 9-10, 8-9, 8, 9, 10 or 11.
  8. 8. The isolated polypeptide of any of claims 1-7, comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of any of SEQ ID NOs 7-10 and 18.
  9. 9. A multimer comprising 60 or more copies of a Head Domain (HD) comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of any of SEQ ID NOs 1-2.
  10. 10. The multimer according to claim 9 comprising 60 or more copies of the isolated polypeptide according to any one of claims 1-8.
  11. 11. A scaffold comprising 60 or more isolated polypeptides according to any one of claims 1-8 on a surface of the scaffold.
  12. 12. The scaffold of claim 11, wherein the isolated polypeptides are all the same polypeptide, or wherein the isolated polypeptides comprise different polypeptides.
  13. 13. A nucleic acid encoding the isolated polypeptide of any one of claims 1-8.
  14. 14. A recombinant expression vector comprising the nucleic acid of claim 13 operably linked to a suitable control sequence.
  15. 15. A recombinant host cell comprising the polypeptide, multimer, scaffold, nucleic acid and/or recombinant expression vector of any of the claims herein.
  16. 16. The nucleic acid of claim 13, wherein the nucleic acid comprises mRNA.
  17. 17. The nucleic acid of claim 16, wherein the mRNA comprises: (a) A 5' cap; (b) Poly (a) tails of 50 to 120 consecutive adenosine residues; (c) A 5' untranslated region comprising a nucleic acid sequence of SEQ ID NO. 12 or 13; (d) A 3' untranslated region comprising one or two copies of a beta-globin mRNA 3' -UTR, optionally wherein the beta-globin mRNA 3' -UTR comprises the nucleic acid sequence of SEQ ID NO:11, and (E) A signal sequence, optionally wherein the signal sequence is located at the N-terminus of the encoded polypeptide, and optionally the signal sequence comprises the amino acid sequence of SEQ ID No. 14.
  18. 18. The nucleic acid of claim 16, wherein the mRNA comprises: (a) A 5' cap; (b) Poly (a) tails of 50 to 120 consecutive adenosine residues; (c) A 5' untranslated region comprising a nucleic acid sequence of SEQ ID NO. 12 or 13; (d) A 3' untranslated region comprising one or two copies of a beta-globin mRNA 3' -UTR, wherein the beta-globin mRNA 3' -UTR comprises the nucleic acid sequence of SEQ ID NO:11, and (E) A signal sequence comprising the amino acid sequence of SEQ ID No. 14 located at the N-terminus of the encoded polypeptide.
  19. 19. A composition comprising a plurality of mrnas encoding 2,3, or 4 polypeptides comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs 5-10 and 18.
  20. 20. A pharmaceutical composition comprising: (a) The polypeptide, multimer, scaffold, nucleic acid, recombinant expression vector and/or cell of any preceding claim, and (B) A pharmaceutically acceptable carrier.

Description

Agents and methods for preventing, treating or limiting henipav infection Description of sequence Listing The computer readable form of the sequence listing is submitted with the present application by electronic submission and is incorporated herein by reference in its entirety. The sequence list is contained in a file created at 2024, 8, 19, with a file name of "23-1410-wo. Xml" and a size of 81,889 bytes. Background Nipah virus (NiV) and Henla virus (HeV) are Henla virus bats-transmitted zoonotics that cause human encephalitis and respiratory symptoms, with mortality between 50% and 100%. NIV was found in 1999, and in the last two decades, bangladesh NIV was almost annually transmitted to humans (strain of bangladesh, NIV-B) and caused epidemic conditions in india (NIV-2) and philippines (strain of malaysia, NIV-2M). Cross-reactive HNV antibodies were detected in African humans and in foxes, highlighting that 20 hundred million people living in areas threatened by HNV extravasation worldwide. To date, there is no approved vaccine or treatment for HNV infection in humans. The HNV genome is a non-segmented single-stranded negative-sense RNA of 18.2kb in length, containing six genes encoding structural proteins N, P, M, F, G and L. HNV entry into host cells requires fusion of the viral membrane and the host plasma membrane by the synergistic action of the attachment (G) glycoprotein and the fusion (F) glycoprotein. The NiV and HeV entry receptors on the surface of host cells are the transmembrane protein tyrosine kinases ephrin-B2 or ephrin-B3. G and F undergo a series of conformational changes upon receptor binding to promote membrane fusion. The NiV G extracellular domain comprises an N-terminal stem (residues 96 to 147), a neck domain (residues 148 to 165), a junction region (residues 166 to 177) and a C-terminal head domain (residues 178 to 602). NiV G forms an interwoven homotetramer of long 200A and wide 120A, and the inter-body disulfide bond formed by the neck and stem is stable. The overall architecture of NiV G employs a unique two-raised and two-lowered configuration. Disclosure of Invention In a first aspect, the invention provides an isolated polypeptide comprising: (a) An attached glycoprotein G Head Domain (GHD) comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of any of SEQ ID NO:1, optionally wherein the head domain comprises the amino acid sequence of SEQ ID NO:1, wherein at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or all 20 of the residues within brackets are the first listed residues, and (B) A multimerization domain capable of producing a multimer comprising 2 to 60 copies of the isolated polypeptide. In one embodiment, the multimerization domain produces 4 copies or 60 copies of the isolated polypeptide. In another embodiment, the multimerization domain comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO. 3-4, optionally wherein the multimerization domain comprises the amino acid sequence of SEQ ID NO. 3, wherein at least 1,2, 3,4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or all 18 of the residues in brackets are the first listed residues. In another embodiment, the polypeptide comprises an amino acid linker between the head domain and the multimerization domain. In another embodiment, the head domain is located at the carboxy terminus of the multimerization domain. In one embodiment, the polypeptide comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:5-6, wherein n is 3-11、4-11、5-11、6-11、7-11、8-11、9-11、10-11、3-10、4-10、5-10、6-10、7-10、8-10、9-10、3-9、4-9、5-9、6-9、7-9、8-9、3-8、4-8、5-8、6-8、7-8、3、4、5、6、7、8、9、10 or 11, optionally wherein n is 8-11, 9-11, 10-11, 8-10, 9-10, 8-9, 8, 9, 10 or 11. In another embodiment, the polypeptide comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of any one of SEQ ID NOs 7-10 and 18. The present disclosure further provides a multimer or scaffold comprising 60 or more copies of a Head Domain (HD) comprising an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of any of SEQ ID NOs 1-2. In one embodiment, the multimer comprises 60 or more copies of an isolated polypeptide of any embodiment or combination of embodiments herein. In various embodiments, the invention provides nucleic acids encoding the isolated polypeptides of any embodiment or combination of embodiments disclosed herein, recombinant expression vectors comprising