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CN-121293297-B - HIV nanoparticle vaccine

CN121293297BCN 121293297 BCN121293297 BCN 121293297BCN-121293297-B

Abstract

The present application relates to an HIV nanoparticle vaccine. The application designs immunogen with stable conformation aiming at MPER and CD4bs respectively through a structural biological strategy and a nanoparticle vaccine platform, and displays the immunogen through virus-like particles (VLPs) respectively, thus finally constructing a combined double epitope vaccine (Cocktail). The vaccine has immune effect in a mouse model, can induce a double-target specific antibody of MPER and CD4bs, and has good broad-spectrum property when exciting the neutralization activity of the multi-subtype HIV pseudovirus.

Inventors

  • ZHANG HUI
  • LIU JIE

Assignees

  • 广州千扬生物医药技术有限公司

Dates

Publication Date
20260505
Application Date
20251204

Claims (9)

  1. The HIV virus gp41 membrane proximal outer region scaffold protein is characterized in that the amino acid sequence is shown as SEQ ID NO. 5.
  2. 2. A recombinant protein is characterized in that the HIV virus gp41 membrane proximal outer region scaffold protein is coupled with ep24 protein with an amino acid sequence shown as SEQ ID NO.6 through linker.
  3. 3. A nanoparticle, characterized by being self-assembled from the recombinant protein of claim 2.
  4. 4. A nanoparticle composition comprising nanoparticle A and nanoparticle B obtained by self-assembly of the recombinant protein according to claim 2, The nanoparticle A is obtained by self-assembling a recombinant protein with an amino acid sequence shown as SEQ ID NO. 27-30 and a Gv-5aa-ep24 recombinant protein with an amino acid sequence shown as SEQ ID NO. 23; or is obtained by self-assembly of recombinant proteins with amino acid sequences shown as SEQ ID NO. 19 and 24-26 and Gv-5aa-ep24 recombinant proteins with amino acid sequences shown as SEQ ID NO. 23.
  5. 5. An HIV vaccine comprising the nanoparticle of claim 3 or the nanoparticle composition of claim 4.
  6. 6. A pharmaceutical combination composition comprising nanoparticle composition 1 and nanoparticle composition 2, The nanoparticle composition 1 is prepared by self-assembling a composition of a recombinant protein with an amino acid sequence shown as SEQ ID NO. 27-30 and a Gv-5aa-ep24 recombinant protein with an amino acid sequence shown as SEQ ID NO. 23 to obtain nanoparticles, and the nanoparticles obtained by self-assembling the recombinant proteins according to claim 2, The nanoparticle composition 2 is prepared by self-assembling a composition of a recombinant protein with an amino acid sequence shown as SEQ ID NO. 19 and 24-26 and a Gv-5aa-ep24 recombinant protein with an amino acid sequence shown as SEQ ID NO. 23, and the nanoparticle prepared by self-assembling the recombinant protein according to claim 2.
  7. 7. A vaccine combination composition comprising nanoparticle composition 1 and nanoparticle composition 2, The nanoparticle composition 1 is prepared by self-assembling a composition of a recombinant protein with an amino acid sequence shown as SEQ ID NO. 27-30 and a Gv-5aa-ep24 recombinant protein with an amino acid sequence shown as SEQ ID NO. 23 to obtain nanoparticles, and the nanoparticles obtained by self-assembling the recombinant proteins according to claim 2, The nanoparticle composition 2 is prepared by self-assembling a composition of a recombinant protein with an amino acid sequence shown as SEQ ID NO. 19 and 24-26 and a Gv-5aa-ep24 recombinant protein with an amino acid sequence shown as SEQ ID NO. 23, and the nanoparticle prepared by self-assembling the recombinant protein according to claim 2.
  8. 8. A biomaterial characterized by being any one of the following: (1) A nucleic acid molecule encoding the HIV virus gp41 membrane proximal outer scaffold protein of claim 1 or the recombinant protein of claim 2; (2) An expression cassette comprising the nucleic acid molecule of (1); (3) A recombinant vector comprising the nucleic acid molecule of (1) or the expression cassette of (2); (4) A recombinant microorganism comprising the nucleic acid molecule of (1), the expression cassette of (2), or the recombinant vector of (3); (5) A cell line comprising the nucleic acid molecule of (1), the expression cassette of (2), or the recombinant vector of (3).
  9. 9. Use of the HIV virus gp41 membrane proximal outer scaffold protein of claim 1, the recombinant protein of claim 2, the nanoparticle of claim 3, the nanoparticle composition of claim 4, the HIV vaccine of claim 5, the drug combination composition of claim 6, the vaccine combination composition of claim 7, or the biomaterial of claim 8 for the preparation of a medicament for the prevention and treatment of HIV.

Description

HIV nanoparticle vaccine Technical Field The application relates to the technical field of HIV vaccines, in particular to an HIV nanoparticle vaccine. Background Human Immunodeficiency Virus (HIV) causes acquired immunodeficiency syndrome (AIDS) by infection of cd4+ T cells, which places a great burden on public health worldwide. Although highly effective antiretroviral therapy (HAART) can delay the course of disease, vaccines are still considered as a fundamental strategy to terminate HIV epidemics. HIV vaccine development has been performed so far for over forty years, and many large clinical trials have failed to provide long lasting, broad spectrum protection, mainly due to high variation of viral surface envelope glycoproteins (Env), glycosylation camouflage barriers, instability of antigen structure, lack of immune pathways inducing bNAbs, and the like. Therefore, how to design accurate immunogens around conserved and functional key epitopes and improve the immune effect by means of a novel platform becomes a hotspot for research of structural vaccines. Based on the differences in HIV-1 genomic sequences, it is largely divided into M groups (main group), N groups (non-main group) and O groups (other groups). Of these, group M is the main group that is popular worldwide, while groups N and O are relatively rare. Group M is the major panel of HIV-1, and contains the majority of HIV-1 strains. Group M is further divided into 9 major subtypes (A-K, where subtype I is absent) which have different epidemic conditions in different regions around the world, where HXB2 is the classical reference strain for subtype B, one of the earliest widely studied HIV-1 strains and 426C is a representative strain for subtype C. In addition to the above subtypes, HIV-1 also exists in a number of recombinant forms (Circulating Recombinant Forms, CRF). Recombinant strains are strains formed by genetic recombination between different subtypes. A number of recombinant forms have been identified worldwide, for example CRF01_AE, CRF02_AG, CRF07_BC, CRF08_BC, and CRF08_BC, respectively, are recombinant from subtype A and subtype E, CRF02_AG, and CRF07_BC, respectively. The HIV-1 envelope protein (Env) is the major antigen on the viral surface and consists of two subunits gp120 and gp 41. Env proteins play a key role in the entry of viruses into host cells. Broad spectrum neutralizing antibodies (broadly neutralizing antibodies, bNAbs) are a class of antibodies capable of neutralizing multiple strains of HIV-1. It is directed mainly to conserved regions on the HIV-1 envelope glycoprotein (Env). Gp120 is responsible for binding to receptors on the surface of host cells, thereby mediating viral entry into the cells. gp120 can be divided into an Outer Domain (OD) and an Inner Domain (ID). gp120-OD contains many epitopes that neutralize antibodies and is an important target for HIV-1 vaccine design. Optimization of bacterially expressed gp120-OD using consensus sequence design, cyclic substitution and structure-directed mutagenesis produced a range of variants with improved yield, biophysical properties, stability and affinity, suitable for targeting a wide range of neutralizing antibody CD4 binding sites (PMID: 30093409). The CD4 binding site (CD 4 bs) is a functional region located in gp120 and is directly involved in binding to the host CD4 receptor. Representative bNAbs such as VRC01 class antibodies target this site. VRC01 is a broad spectrum neutralizing antibody derived from HIV-infected individuals, isolated from U.S. NIH VACCINE RESEARCH CENTER (VRC), targeting the gp120 CD4 binding site (CD 4 bs), and is capable of neutralizing most HIV-1 strains. The CD4bs region, while relatively conserved, is surrounded by multiple N-linked glycosylation sites (e.g., N276, N197, N301, etc.) at its periphery, creating a steric barrier that makes routine immunization difficult to elicit antibodies against this epitope. Thus, in the design of CD4bs vaccines, partial removal of interfering sugar groups to expose critical amino acids, while preserving epitope conformation, is a design focus. Among them, the best known antigens for vaccines based on CD4bs are eOD-GT8.eOD-GT8 is an HIV-1 gp120 ectodomain artificial immunogen designed by IAVI (International AIDS vaccine initiative organization) in combination with Stokes institute, aimed at specifically activating VRC 01-type broad-spectrum neutralizing antibody precursor B cells by means of a germ-line targeting strategy. The method adopts an optimized ectodomain and glycosyl masking strategy, improves the affinity and specificity to target B cells, enters a phase 1 clinical test, is used for evaluating the safety and immunogenicity in healthy adults, and provides an important theoretical and experimental basis for developing HIV vaccines (PMID: 26089355). The membrane proximal outer region (Membrane Proximal External Region, MPER) is positioned at the front end of the gp41 transmembrane region,