AEGiS-13IAC: Identification, characterization and synthesis of lysozyme mimetics with potent anti-HIV activity.

13th International AIDS Conference

Durban, South Africa - July 9-July 14, 2000

Identification, characterization and synthesis of lysozyme mimetics with potent anti-HIV activity.

Int Conf AIDS 2000 Jul 9-14; 13:(abstract no. TuOrA405)

Huang P, Sun Y, Chen H-C, Huang P, Lee-Huang S
P. Huang, American BioSciences, 8 Whittier Place, #7E, Boston, MA 02114, United States, E-mail: p.huang@rcn.com

BACKGROUND: Lysozyme acts against bacterial/viral infection, tumor growth, and modulates host immunity. These combined effects distinguish lysozyme from other drugs. Recently, it was discovered that lysozyme contributes to the anti-HIV activity of b-core of hCG. This opens the way to an entirely new class of anti-AIDS medicines based on mimicking the activity of lysozyme. Our goal is to investigate how lysozyme can best be used as an anti-HIV drug.

METHODS: Rational fragmentation, activity mapping and peptide synthesis were used. Fragments generated were analyzed by HPLC. Identity of the fragments was determined by amino acid sequencing. Anti-viral activity was measured by inhibition on HIV-1.

RESULTS: Limited proteolysis of human lysozyme resulted in ten fragments. Anti-HIV activity was found in a 18-amino acid fragment (HL18) with the sequence RVVRDPQGIRAWVAWRNR. HL18 exhibits comparable anti-HIV activity to intact lysozyme, with 50% inhibition concentration (IC50) at 58-68 nM. Further mapping and modeling generated a derivative of 9 amino acids (HL9) with the sequence RAWVAWRNR. Here, we report that HL9 is potent against a wide spectrum of HIV-1, including primary isolates, laboratory strains and resistant strains with IC50 of 55-68 nM. No toxicity was observed in the dose range of the assay. HL9 also inhibits the proliferation of HHV8 infected cells of AIDS patients with Kaposi's sarcoma. The antiviral activity was confirmed by synthetic peptide with identical sequence. Peptides with scrambled sequences are inactive. Substitution of the R with K, N or Q all result in diminution of the antiviral activity.

CONCLUSION: This is the first identification and synthesis of an anti-HIV mimetic from human lysozyme. Our results suggest the HL9 is a promising candidate as a novel anti-AIDS drug. It is derived from a human protein, it has potent activity against a broad spectrum of HIV isolates, it can be easily synthesized and is readily available.

Keywords: AEGIS, HIV, HIV Infections, Anti-HIV Agents, HIV-1, Muramidase, HIV Seropositivity, Acquired Immunodeficiency Syndrome, Amino Acid Sequence, HIV Protease, HIV Long Terminal Repeat, HIV Integrase, HIV-1 Reverse Transcriptase, HIV Core Protein p24, Chorionic Gonadotropin, Human, chemical synthesis, genetics, immunology
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TuOrA405