Griffithsin, glycosylation-resistant griffithsin, and related conjugates, compositions, nucleic acids, vectors, host cells, methods of production and methods of use
Inventors
Boyd, Michael R. • Mori, Toshiyuki • O'Keefe, Barry R.
Assignees
US Department of Health and Human Services
Publication Number
US-8394764-B2
Publication Date
2013-03-12
Expiration Date
2025-05-27
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Abstract
An isolated and purified nucleic acid molecule that encodes a polypeptide comprising at least eight contiguous amino acids of SEQ ID NO: 3, wherein the at least eight contiguous amino acids have anti-viral activity, as well as an isolated and purified nucleic acid molecule that encodes a polypeptide comprising at least eight contiguous amino acids of SEQ ID NO: 3, wherein the at least eight contiguous amino acids have anti-viral activity, and, when the at least eight contiguous amino acids comprise amino acids 1-121 of SEQ ID NO: 3, the at least eight contiguous amino acids have been rendered glycosylation-resistant, a vector comprising such an isolated and purified nucleic acid molecule, a host cell comprising the nucleic acid molecule, optionally in the form of a vector, a method of producing an anti-viral polypeptide or conjugate thereof, the anti-viral polypeptide itself, a conjugate or fusion protein comprising the anti-viral polypeptide, and compositions comprising an effective amount of the anti-viral polypeptide or conjugate or fusion protein thereof. Further provided are methods of inhibiting prophylactically or therapeutically a viral infection of a host.
Core Innovation
The invention provides an isolated and purified nucleic acid molecule that encodes a polypeptide comprising at least eight contiguous amino acids of SEQ ID NO: 3, wherein these amino acids have anti-viral activity. This includes nucleic acids encoding a polypeptide comprising amino acids 1-121 of SEQ ID NO: 3 rendered glycosylation-resistant, vectors comprising such nucleic acids, host cells transformed therewith, methods of producing anti-viral polypeptides or conjugates, the anti-viral polypeptides or conjugates themselves, and compositions containing effective amounts of these polypeptides or conjugates. Methods of inhibiting viral infections prophylactically or therapeutically by administering these anti-viral polypeptides or conjugates to a host are also provided.
The problem addressed by the invention stems from limitations in treating and preventing infections by viruses such as HIV and influenza. Existing anti-retroviral agents like AZT suffer from toxicity and limited therapeutic indices, with a need for agents acting early in the viral replicative cycle to inhibit infection and transmission. Preventive means against HIV, particularly via heterosexual transmission, lack effective vaccines and require novel strategies such as topical microbicides. Influenza infection remains a major cause of illness and mortality with limited treatment or preventive options due to viral mutation. There is a critical need for new broad-spectrum anti-viral agents that are virus-specific, potent against diverse strains, stable under physiological and environmental conditions, and readily produced.
The invention overcomes these challenges by providing griffithsin, an anti-viral polypeptide isolated from Griffithsia sp., possessing broad-spectrum activity against viruses including retroviruses and influenza virus. Griffithsin binds to high mannose oligosaccharide moieties on viral envelope glycoproteins such as gp120, thereby inhibiting viral binding, fusion, and entry. The invention contemplates variants, fragments, fusion proteins, and conjugates of griffithsin, including glycosylation-resistant forms, and provides nucleic acid sequences and expression systems for large-scale production. The use of griffithsin and its derivatives as prophylactic and therapeutic agents, including delivery via recombinant host cells, topical applications, and as components of antiviral compositions and conjugates, addresses the shortcomings in existing antiviral therapeutics and prophylactics.
Claims Coverage
The independent claims relate to methods of inhibiting viral infection in a host by administering an effective amount of anti-viral polypeptides or conjugates comprising specific sequences with glycosylation-resistant modifications.
Use of glycosylation-resistant antiviral polypeptides to inhibit viral infections
A method involving administration of an anti-viral polypeptide or conjugate comprising SEQ ID NO:3 or a variant thereof where amino acid residues at positions 45, 60, 71, or 104 are rendered glycosylation-resistant, providing anti-viral activity to inhibit viral infection in a host.
Topical administration for influenza viral infection treatment
Administration of the glycosylation-resistant antiviral polypeptide or conjugate topically to the host, specifically to the respiratory system, including administration as an aerosol or microparticulate powder, for treating influenza viral infection.
Use against retroviral infections including human immunodeficiency virus
Application of the anti-viral polypeptide or conjugate comprising SEQ ID NO:3 or SEQ ID NO:2 for inhibiting retroviral infections, notably human immunodeficiency viral infections such as HIV-1 and HIV-2.
The claims cover methods of prophylactic or therapeutic viral infection inhibition by administering anti-viral polypeptides or conjugates comprising SEQ ID NO:3 or SEQ ID NO:2, including glycosylation-resistant variants, with specific emphasis on topical administration for influenza and retroviral infections, notably HIV.
Stated Advantages
The invention provides anti-viral agents that act early in the viral replicative cycle.
The antiviral agents are virus-specific and potent against a broad spectrum of viral strains and isolates.
The agents are stable under physiological and environmental conditions and can be readily and inexpensively produced.
Griffithsin possesses broad anti-retroviral and anti-influenza activity, including virucidal effects and inhibition of viral entry.
Recombinant production methods enable large-scale, efficient manufacturing of active polypeptides.
Agents can be delivered via various routes including topical, inhalation, and parenteral, enabling versatile therapeutic and prophylactic use.
Documented Applications
Prophylactic and therapeutic inhibition of viral infections in hosts, specifically retroviruses including HIV-1 and HIV-2, and influenza virus.
Topical microbicides to prevent sexual transmission of HIV infection by application to genital mucosa.
Therapeutic and prophylactic treatment of influenza infections, including aerosol administration to the respiratory system.
Use of recombinant host cells, including genetically engineered lactobacilli, for in vivo delivery of griffithsin to mucosal surfaces.
Formulations and compositions for antiviral treatment administered orally, topically, parenterally, or via other routes to prevent or treat viral infections.
Removal or inactivation of infectious virus from biological samples such as blood products, tissues, or fluids via griffithsin coupled to solid supports (e.g., magnetic beads).
Induction of immune responses using compositions comprising naturally-occurring non-infectious virus treated with griffithsin for vaccine or immunotherapy applications.
Methods for producing anti-griffithsin antibodies that can be used for detection, purification, or removal of viruses or infected cells.
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