Search

BR-112018072248-B1 - Modified HIV-1 envelope GP140 protein, its use, and composition of an HIV-1 vaccine.

BR112018072248B1BR 112018072248 B1BR112018072248 B1BR 112018072248B1BR-112018072248-B1

Abstract

The present invention provides immunogens for HIV-1 vaccine. Some of the immunogens contain a soluble gp140-derived protein carrying a modified N-terminus of the HR1 region in gp41. Some of the immunogens contain a trimeric Env-derived HIV-1 protein presented on an 8-nanoparticle platform. The invention also provides methods for using the HIV-1 vaccine immunogens to elicit an immune response or treat HIV infections.

Inventors

  • Leopold Kong
  • IAN A. WILSON
  • Natalia De Val
  • ANDREW B. WARD
  • Dennis Burton
  • Linling He
  • Jiang Zhu

Assignees

  • THE SCRIPPS RESEARCH INSTITUTE

Dates

Publication Date
20260317
Application Date
20170501
Priority Date
20160502

Claims (20)

  1. 1. Modified HIV-1 envelope gp140 protein, characterized in that it comprises a gp120 polypeptide and a gp41 polypeptide, wherein the N-terminal amino acid residues 548-568 of the heptad 1 (HR1) region of the gp41 polypeptide are replaced by a loop sequence of 6 to 14 amino acid residues in length that stabilizes the pre-fusion gp140 structure, wherein the numbering of the amino acid residues corresponds to the HxB2 nomenclature.
  2. 2. Modified HIV-1 envelope protein gp140, according to claim 1, characterized in that the gp41 polypeptide is gp41ECTO.
  3. 3. Modified HIV-1 envelope protein gp140, according to claim 1, characterized in that the gp120 and gp41 polypeptides are from different HIV-1 strains.
  4. 4. Modified HIV-1 envelope gp140 protein, according to claim 1, characterized in that it is derived from the HIV-1 strain BG505.
  5. 5. Modified HIV-1 envelope gp140 protein according to claim 1, characterized in that the loop sequence comprises (GS)n (SEQ ID NO: 23), wherein n is an integer between 3 and 7 inclusive.
  6. 6. Modified HIV-1 envelope gp140 protein, according to claim 1, characterized in that the loop sequence comprises (GS)4 (SEQ ID NO: 24).
  7. 7. Modified HIV-1 envelope gp140 protein, according to claim 1, characterized in that the loop sequence comprises 10 amino acid residues.
  8. 8. Modified HIV-1 envelope gp140 protein, according to claim 7, characterized in that the loop sequence comprises any of the SEQ ID Nos: 1-5.
  9. 9. Modified HIV-1 envelope gp140 protein, according to claim 1, characterized in that the loop sequence comprises 8 amino acid residues.
  10. 10. Modified HIV-1 envelope gp140 protein according to claim 9, characterized in that the loop sequence comprises any one of the SEQ ID Nos: 6-10.
  11. 11. Modified HIV-1 envelope gp140 protein according to claim 1, characterized in that it further comprises a flexible linker sequence comprising (G4S)2 (SEQ ID NO: 22) or SGS, which replaces residues 508-511 in the cleavage site, between gp120 and gp41.
  12. 12. Modified HIV-1 envelope gp140 protein, according to claim 1, characterized in that it further comprises a linker sequence comprising the sequence set forth in any of the SEQ ID Nos: 16-20 which replaces residues 501-518 in the cleavage site, between gp120 and gp41, and wherein the numbering of the amino acid residues corresponds to that of the HIV-1 strain BG505. SOSIP.664 gp140.
  13. 13. Modified HIV-1 envelope gp140 protein according to claim 1, characterized in that it further comprises an engineered disulfide linkage between gp120 and gp41, between residues A501C and T605C.
  14. 14. Modified HIV-1 envelope gp140 protein according to claim 1, characterized in that it comprises a gp140 trimer with each monomer comprising a gp120 polypeptide and a gp41ECTO polypeptide, wherein the gp41ECTO polypeptide is derived from the BG505 strain of HIV-1, and wherein the N-terminus of the heptad 1 (HR1) region (SEQ ID No: 28) in the gp41ECTO polypeptide is replaced by a loop sequence as shown in SEQ ID No: 6.
  15. 15. Modified HIV-1 envelope gp140 protein according to claim 14, characterized in that it further comprises (a) a (G4S)2 linker sequence (SEQ ID NO: 22) that replaces residues 508-511 in the cleavage site and (b) an engineered disulfide bond between residues A501C and T605C.
  16. 16. HIV-1 vaccine composition, characterized in that it comprises an HIV-1 Env-derived trimer immunogen presented in a self-assembling nanoparticle or a virus-like particle (VLP), wherein the HIV-1 Env-derived trimer immunogen comprises a gp120 polypeptide and a gp41 polypeptide, as defined in any one of claims 1 to 15, wherein the N-terminal amino acid residues 548-568 of the heptad 1 (HR1) region of the gp41 polypeptide are replaced by a loop sequence of 6 to 14 amino acid residues in length that stabilizes the pre-fusion gp140 structure, wherein the numbering of the amino acid residues corresponds to the HxB2 nomenclature.
  17. 17. HIV-1 vaccine composition according to claim 16, characterized in that the HIV-1 Env-derived trimer immunogen is V1V2, gp120 or gp140.
  18. 18. HIV-1 vaccine composition according to claim 16, characterized in that the HIV-1-derived immunogenic trimer Env is a modified gp140 protein comprising a gp120 polypeptide and a gp41ECTO polypeptide, and wherein the N-terminal of the heptad 1 (HR1) region of the gp41ECTO polypeptide is replaced by a loop sequence of about 6 to about 14 amino acid residues that stabilize the structure of the pre-fusion gp140.
  19. 19. HIV-1 vaccine composition according to claim 18, characterized in that the loop sequence comprises (a) (GS)n (SEQ ID NO:23), wherein n is any integer between 3 and 7 inclusive, or (b) a sequence rationally redesigned by means of ensemble-based protein design.
  20. 20. HIV-1 vaccine composition according to claim 18, characterized in that the modified gp140 protein is covalently fused to the nanoparticle platform.

Description

CROSS-REFERENCE TO RELATED REQUESTS [001] This patent application claims the benefit of priority to U.S. Provisional Patent Application No. 62/330,604 (filed May 2, 2016). The full description of the priority application is incorporated herein by reference in its entirety and for all purposes. GOVERNMENT STATEMENT OF SUPPORT [002] The invention was made with government support under AI100663 and AI084817 granted by the National Institutes of Health. The government has certain rights in the invention. BACKGROUND OF THE INVENTION [003] Human immunodeficiency virus type 1 (HIV-1) is the leading cause of acquired immunodeficiency syndrome (AIDS), which is considered one of the world's major health problems. It is an RNA virus of the Retroviridae family. The HIV-1 genome encodes at least nine proteins that are divided into three classes: the main structural proteins Gag, Pol, and Env; the regulatory proteins Tat and Rev; and the accessory proteins Vpu, Vpr, Vif, and Nef. HIV-1 can be divided into several different clades, for example, A, B, C, D, E, F, G, H, J, and K, which vary in prevalence worldwide. Each clade comprises different strains of HIV-1 that have been grouped together based on their genetic similarity. [004] The initial phase of the HIV-1 replicative cycle involves the virus binding to susceptible host cells followed by fusion of viral and cellular membranes. These events are mediated by outer viral envelope glycoproteins that are first synthesized as a fusion-incompetent precursor envelope glycoprotein (Env) known as gp160. The genetic diversity of HIV-1 makes it extremely difficult to develop an effective vaccine against strains for multiple HIV-1 clades. Great efforts have been made in the last two decades to produce a preventive HIV vaccine. Although several candidate vaccines have been developed, they have failed to prevent HIV-1 infection in clinical trials. [005] Generating an antibody response capable of neutralizing a wide range of clinical isolates remains a major challenge in vaccine development for human immunodeficiency virus type 1 (HIV-1). There is a strong and urgent need for a vaccine that is safe and effective worldwide. The present invention addresses this and other unmet needs in the art. SUMMARY OF THE INVENTION [006] In one aspect, the invention provides modified HIV-1 envelope gp140 proteins. The proteins are composed of a gp120 polypeptide and a gp41 polypeptide, with the N-terminus of the heptad 1 (HR1) region of the gp41 polypeptide being replaced by a loop sequence of about 6 to about 14 amino acid residues in length that stabilizes the pre-fusion gp140 structure. In some of these proteins, the gp41 polypeptide is gp41ECTO. Preferably, the modified HIV-1 gp140 protein is a trimer. In some embodiments, the gp120 polypeptide and the gp41 polypeptide are derived from the same HIV-1 strain or subtype. For example, both the gp120 polypeptide and the gp41 polypeptide in the modified HIV-1 gp140 protein may be derived from the HIV-1 BG505 strain. In some embodiments, the gp120 polypeptide and the gp41 polypeptide are derived from different strains or subtypes of HIV-1. For example, a gp41 polypeptide engineered from the BG505 strain of HIV-1, as exemplified here, can be used to form chimeric gp140 immunogens with a gp120 polypeptide derived from many other strains or subtypes of HIV-1. [007] In some modified HIV-1 envelope gp140 proteins of the invention, the loop sequence contains (GS)n (SEQ ID No.: 23), with n being any integer between 3 and 7 inclusive. In some of these embodiments, the loop sequence is (GS)4 (SEQ ID No.: 24). In some embodiments, the loop sequence is obtained through novel rational design. In some of these embodiments, the loop sequence is obtained through ensemble-based protein design. In some modified HIV-1 gp140 proteins of the invention, the loop sequence contains 10 amino acid residues. Some examples of these loop sequences are shown in SEQ ID Nos.: 1-5. In some modified HIV-1 gp140 proteins, the loop sequence contains 8 amino acid residues. Some examples of these loop sequences are shown in SEQ ID Nos.: 6-10. [008] In some embodiments, the modified HIV-1 gp140 proteins of the invention further contain a flexible linker sequence that replaces the cleavage site sequence between gp120 and gp41. In some of these embodiments, the linker sequence has a (G4S)2 (SEQ ID No.: 22) or SGS sequence, which replaces residues 508-511 in the cleavage site. In some other embodiments, the linker sequence contains 8 amino acid residues and replaces residues 501-518 in the cleavage site. In these embodiments, the numbering of the amino acid residues corresponds to that of the HIV-1 BG505 strain. SOSIP.664 gp140. In some exemplified proteins, the linker sequence contains the sequence shown in any of the SEQ ID Nos.: 16-20. [009] In some embodiments, the modified HIV-1 gp140 proteins of the invention further contain (a) an engineered disulfide bond between gp120 and gp41 and