HIV-1 Env fusion peptide nanoparticle carrier conjugates and their use
Inventors
Kwong, Peter • Ou, Li • Mascola, John • Zhang, Baoshan • Xu, Kai • Kong, Wing-pui • Tsybovsky, Yaroslav • Chao, Cara W. • Boyington, Jeffrey • WANG, Yiran
Assignees
US Department of Health and Human Services
Publication Number
US-12053519-B2
Publication Date
2024-08-06
Expiration Date
2039-09-23
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Abstract
Embodiments of immunogenic conjugates including the HIV-1 Env fusion peptide and methods of their use and production are disclosed. In several embodiments, the immunogenic conjugates can be used to generate an immune response to HIV-1 Env in a subject, for example, to treat or prevent an HIV-1 infection in the subject.
Core Innovation
This disclosure provides novel immunogenic conjugates comprising HIV-1 Env fusion peptides conjugated to a self-assembling protein nanoparticle carrier, designed to elicit an immune response to HIV-1 Env in a subject. The self-assembling protein nanoparticle carrier is a multimer comprising fusion proteins, each fusion protein incorporating a self-assembling nanoparticle subunit fused to a heterologous carrier protein. The fusion proteins self-assemble into the nanoparticle carrier onto which HIV-1 Env fusion peptides are conjugated to form immunogenic conjugates usable for priming an immune response against HIV-1 Env fusion peptide epitopes.
The challenge addressed by the invention stems from the global HIV-1 pandemic, where millions are infected yearly despite available antiretroviral therapies. HIV-1 uses protective mechanisms, such as the Env spike composed of gp120 and gp41 proteins, which evade humoral recognition. A significant difficulty in HIV-1 vaccine development is focusing the immune response to target epitopes that elicit broadly neutralizing antibodies. Prior efforts to develop immunogens capable of inducing such responses have not fully met the need.
The invention overcomes this challenge by constructing immunogens that elicit immune responses specifically to the HIV-1 Env fusion peptide using multivalent self-assembling protein nanoparticle carriers conjugated with the fusion peptides. The carriers may also incorporate heterologous T-cell helper epitopes to enhance immunogenicity. The conjugates can generate neutralizing immune responses to HIV-1 Env, providing potential vaccines and diagnostic tools for detecting target antibodies in a polyclonal serum response.
Claims Coverage
The patent claims cover two independent aspects: the immunogenic conjugate comprising the self-assembling protein nanoparticle carrier conjugated to HIV-1 Env fusion peptides, and recombinant self-assembling nanoparticle subunits with engineered cysteine substitutions to enhance nanoparticle stability.
Immunogenic conjugate with HIV-1 Env fusion peptides
An immunogenic conjugate comprising a self-assembling protein nanoparticle carrier formed from a multimer of fusion proteins, each fusion protein comprising a self-assembling protein nanoparticle subunit fused to a heterologous carrier protein. HIV-1 Env fusion peptides comprising from the N-terminus a sequence starting at residue 512 to one of residues 514-521 of an HIV-1 Envelope protein (HXB2 numbering) are conjugated to the nanoparticle carrier. The conjugate elicits an immune response to HIV-1 Env.
Self-assembling nanoparticle subunits with stabilizing cysteine substitutions
Recombinant self-assembling nanoparticle subunits comprising cysteine substitutions that introduce one or more non-native disulfide bonds to increase nanoparticle stability. The cysteine substitutions listed include: 121C and 131C (or variants) in lumazine synthase; 53C and 94C, or 53C and 96C, or 146C and 185C in encapsulin; T81C, 53C and 100C, or 82C and 80C in Acinetobacter phage AP205; and 25C and 127C, 14C and 36C, 29C and 127C, 18C and 36C substitutions in Hepatitis B capsid protein, as per specified reference sequences.
Self-assembling nanoparticle subunits of defined sequences
Recombinant self-assembling nanoparticle subunits consisting of amino acid sequences of ferritin, lumazine synthase, encapsulin, Acinetobacter phage AP205, or Hepatitis B capsid protein as set forth in the SEQ ID NOs provided, suitable for use in forming nanoparticle carriers.
Fusion of self-assembling nanoparticle subunits to heterologous carrier proteins
The recombinant self-assembling nanoparticle subunits can be fused to heterologous carrier proteins selected from tetanus toxin heavy chain C fragment, diphtheria toxin variant CRM197, H. influenzae protein D, Keyhole Limpet Hemocyanin functional unit, Meningococcal outer membrane protein complex, Outer-membrane lipoprotein carrier protein, or Cholera toxin B subunit to form nanoparticle carriers.
The claims define immunogenic conjugates composed of self-assembling protein nanoparticle carriers conjugated with HIV-1 Env fusion peptides capable of eliciting immune responses, and recombinant nanoparticle subunits engineered for enhanced stability via cysteine substitutions, including their nucleic acids, nanoparticle assemblies, conjugated vaccines, compositions, and methods of immune response induction.
Stated Advantages
The immunogenic conjugate provides a multivalent platform with superior binding capability for engaging HIV-1 Env fusion peptide-directed broadly neutralizing antibodies.
Use of the self-assembling protein nanoparticle carrier enhances priming of the immune response to HIV-1 Env fusion peptides compared to monomeric carrier conjugates.
Incorporation of heterologous T-cell helper epitopes in the fusion proteins further improves the immune response elicited by the immunogenic conjugate.
Introducing non-native disulfide bonds in nanoparticle subunits stabilizes the assembled nanoparticles, increasing resistance to disassembly.
Documented Applications
The immunogenic conjugates can be used to generate an immune response to HIV-1 Env in a subject, for example, to treat or prevent HIV-1 infection in the subject.
The immunogenic conjugates and compositions comprising same can be used in coordinate (prime-boost) immunization protocols or combinatorial formulations for prophylactic or therapeutic purposes against HIV-1.
The disclosed immunogenic conjugates are useful as potential vaccines for HIV-1, capable of eliciting neutralizing immune responses including antibodies targeting the HIV-1 Env fusion peptide.
These conjugates can also be used as diagnostic molecules to detect and quantify HIV-1 Env fusion peptide-specific antibodies in a polyclonal serum response.
The recombinant self-assembling nanoparticles conjugated to vaccine antigens can be administered to subjects to generate an immune response against vaccine antigens.
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