Bifunctional molecules for inhibiting HIV entry
Disclosed herein are bifunctional molecules which inhibit HIV entry into the target cell. Also disclosed are novel anti-HIV therapeutics for treatment of patients infected by HIV, including non-B and multi-drug resistant strains.
1. A bifunctional protein consisting essentially of:
a first C-terminal heptad repeat (CHR) peptide of a human immunodeficiency virus 1 (HIV-1) gp41 containing a pocket-binding domain and a heptad repeat (HR)-binding domain, wherein said first CHR peptide is directly linked to;
a flexible linker comprising the amino acid sequence (GGGGS) n , wherein n is an integer between 6 and 8 (SEQ ID NOs:39, 40 and 41), which in turn is directly linked to;
a second CHR peptide of an HIV-1 gp41 containing a HR-binding domain and a tryptophan-rich domain.
2. The bifunctional protein of claim 1 wherein said first CHR peptide comprises the amino acid sequence selected from the group consisting of C34 (SEQ ID NO:11), C36 (SEQ ID NO:12), C38 (SEQ ID NO:3), C46 (SEQ ID NO:13), T1144 (SEQ ID NO:4), T1144-C10 (SEQ ID NO:5), sifuvirtide (SEQ ID NO:14), C35-EK (SEQ ID NO:15), CP621-652 (SEQ ID NO:16), CP32M (SEQ ID NO:17), T1249 (SEQ ID NO:18), PBD-4HR (SEQ ID NO:19), and C36B (SEQ ID NO:20).
3. The bifunctional protein of claim 1 wherein said second CHR peptide comprises the amino acid sequence selected from the group consisting of T20 (SEQ ID NO:2), T20-A (SEQ ID NO:22), and 4HR-LBD (SEQ ID NO:23).
4. The bifunctional protein of claim 1 additionally comprising the amino acid sequence YSSGRIVTD (SEQ ID NO:53) or NSSGRIVTD (SEQ ID NO:42) at the C-terminus of said bifunctional protein.
5. The bifunctional protein of claim 1 wherein said bifunctional protein comprises the amino acid sequence of GPLGSTTWEAWDRAIAEYAARIEALLRALQEQQEKNEAALREL(GGGGS) 7 YTSLIHSLIEESQN QQEKNEQELLELDKWASLWNWFYSSGRIVTD (SEQ ID NO:6).
6. The bifunctional protein of claim 1 wherein said bifunctional protein comprises the amino acid sequence of GPLGSTTWEAWDRAIAEYAARIEALLRALQEQQEKNEAALREL(GGGGS) 7 YTSLIHSLIEESQN QQEKNEQELLELDKWASLWNWFNSSGRIVTD (SEQ ID NO:7).
7. The bifunctional protein of claim 1 wherein said bifunctional protein comprises the amino acid sequence of GPLGSTTWEAWDRAIAEYAARIEALLRALQEQQEKNEAALREL(GGGGS) 6 YTSLIHSLIEESQN QQEKNEQELLELDKWASLWNWF (SEQ ID NO:8).
8. The bifunctional protein of claim 1 wherein said bifunctional protein is produced by recombinant DNA technology.
9. The bifunctional protein of claim 8 wherein said bifunctional protein is produced in an expression system selected from the group consisting of bacteria, yeast, insect cells and mammalian cells.
10. The bifunctional protein of claim 9 wherein said bifunctional protein is produced in Escherichia coli.
11. The bifunctional protein of claim 1 wherein said bifunctional protein is synthesized on a solid or in solution.
12. The bifunctional protein of claim 1 wherein said bifunctional protein is synthesized as several separated segments and then connected together.
13. A method of inhibiting human immunodeficiency virus infection comprising: administering a bifunctional protein of claim 1 to an individual infected with the human immunodeficiency virus, thereby inhibiting entry of the virus into a target cell, and blocking infection of the target cell with the virus.
14. The bifunctional protein of claim 1 comprising the amino acid sequence of TTWEAWDRAIAEYAARIEALLRALQEQQEKNEAALREL(GGGGS) 7 YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNWF (SEQ ID NO:25).
15. The bifunctional protein of claim 1 comprising the amino acid sequence of TTWMEWDREINNYTSLIHSLIEESQNQQEKNEQELLEL(GGGGS) 7 YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNWF (SEQ ID NO:31).
16. The bifunctional protein of claim 1 comprising the amino acid sequence of TTWMEWDREINNYTSLIHSLIEESQNQQEKNEQELLEL(GGGGS) 7 YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNWF (SEQ ID NO:33).