Stabilized therapeutic small helical antiviral peptides
Provided are constrained peptides that inhibit HIV assembly. Pharmaceutical compositions comprising the above peptides are also provided. Additionally provided are methods of inhibiting replication of a capsid-containing virus in a cell. Also provided are methods of treating a mammal infected with a capsid-containing virus. Further provided are methods of treating a mammal at risk for infection with a capsid-containing virus. Methods of making the above peptides are additionally provided, as are uses of the above peptides and pharmaceutical compositions.
1. An isolated peptide from 10 to 23 amino acids long, wherein the sequence of the amino acids of the isolated peptide comprises (I/L/V)(T/S/A/V/C)(F/I/L/V/Y/M/W)(D/E/S)(D/E)(L/F/I/V/Y/M/W)(L/D/T/F/I/V/Y/M/W)(D/E/A/S)(Y/F/I/L/V/M/W)(Y/F/I/L/V/M/T) (SEQ ID NO:2), wherein two of the amino acids are unnatural amino acids having either R or S stereochemistry at the α-carbon, wherein the unnatural amino acids are each (S)-α-2-(4′-pentenyl)alanine or (S)-α-2-(7′-octenyl)alanine,
wherein the α-carbon of the unnatural amino acids comprises a methyl group and an olefinic group, where the two olefinic groups of the unnatural amino acids are on the same side of the α-helix and are joined to form a cross-link between the two unnatural amino acids,
wherein the two unnatural amino acids replace two of the amino acids at any positions selected from the group consisting of 4 amino acids apart (i and i+4) and 7 amino acids apart (i and i+7); and
wherein the cross-link between the two unnatural amino acids is a C1-C10 alkyl, alkenyl, alkynyl, or (R1-K-R1) n ; each of which is substituted with a 0-6 R2,
wherein R1 is an alkyl, alkenyl, or alkynyl;
K is O, S, SO, SO 2 , CO, CONR4, or
R2 is a halo, C1-C10 alkyl, OR3, N(R3) 2 , SR3SOR3, SO 2 R3, CO 2 R3, R3, a fluorescent moiety, or a radioisotope;
R3 is H or a C1-C10 alkyl;
R4 is H, alkyl, or a therapeutic agent; and
n is an integer from 1-4.
2. The peptide of claim 1 , wherein the sequence of the peptide comprises (I/V)(T/S)(F/W/Y)(E/S)(D/E)L(L/D/T)(D/A/S)(Y/F)(Y/M) (SEQ ID NO:3).
3. The peptide of claim 1 , comprising 11 to 23 amino acids, wherein the amino acid following (Y/F/I/L/V/M/T) is (G/S/T/N/H/C/L/R/D/E/Q/M/K) (SEQ ID NO:4).
4. The peptide of claim 3 , wherein the sequence of the peptide comprises SEQ ID NO:8.
5. The peptide of claim 3 , comprising 12 to 23 amino acids, wherein the amino acid following (G/S/T/N/H/C/L/R/D/E/Q/M/K) is (P/M/R/K) (SEQ ID NO:5).
6. The peptide of claim 5 , wherein the sequence of the peptide comprises SEQ ID NO:9.
7. The peptide of claim 6 , wherein the sequence of the peptide comprises ITFXDLLXYYGP (SEQ ID NO:6) and wherein X represents the unnatural amino acids.
8. The peptide of claim 1 , wherein the cross-link between the two unnatural amino acids is
wherein the (C)s are the α-carbons of the unnatural amino acids.
9. The peptide of claim 1 , wherein the peptide comprises SEQ ID NO:10.
10. The peptide of claim 3 , wherein the peptide comprises SEQ ID NO:11.
11. The peptide of claim 5 , wherein the peptide comprises SEQ ID NO:12.
12. The peptide of claim 1 , comprising
wherein the (C)s are the α-carbons of the unnatural amino acids.
13. The peptide of claim 1 , consisting of
wherein the (C)s are the α-carbons of the unnatural amino acids.
14. The peptide of claim 1 , further comprising a detectable moiety, a therapeutic compound, or an antigen.
15. The peptide of claim 14 , wherein the detectable moiety is a fluorescent moiety.
16. The peptide of claim 14 , wherein the detectable moiety is a radioactive moiety.
17. The peptide of claim 14 , further comprising an antigen.
18. The peptide of claim 17 , wherein the antigen is a human immunodeficiency virus antigen.
19. The peptide of claim 14 , further comprising a therapeutic compound.
20. The peptide of claim 19 , wherein the therapeutic compound comprises an oligopeptide less than 20 amino acids long.
21. The peptide of claim 19 , wherein the therapeutic compound comprises an oligopeptide less than 10 amino acids long.
22. The peptide of claim 19 , wherein the therapeutic compound is an organic compound.
23. The peptide of claim 22 , wherein the organic compound is an antiviral compound.
24. The peptide of claim 19 , wherein the therapeutic compound is bound to the rest of the peptide with an ester bond susceptible to cleavage by a cellular esterase.
25. A pharmaceutical composition comprising the isolated peptide of claim 1 , in a pharmaceutically acceptable carrier.
26. A method of inhibiting replication of a human immunodeficiency virus in a cell, the method comprising contacting the cell with the peptide of claim 1 in a manner sufficient to inhibit replication of the capsid-containing virus in the cell.
27. The method of claim 26 , wherein the cell is in a mammal infected with the human immunodeficiency virus.
28. The method of claim 27 , wherein the mammal is a human.
29. The method of claim 27 , further comprising treating the mammal with at least one additional anti-viral treatment.
30. A method of treating a mammal infected with a human immunodeficiency virus, the method comprising administering the pharmaceutical composition of claim 25 to the mammal in a manner sufficient to treat the mammal.
31. The method of claim 30 , wherein the mammal is a human.
32. The method of claim 30 , wherein the peptide is
wherein the (C)s are the α-carbons of the unnatural amino acids.
33. The method of claim 30 , wherein the mammal is a pregnant female.
34. The method of claim 30 , further comprising treating the mammal with at least one additional anti-viral treatment.