Autophagy-inducing peptide analogs
An autophagy-inducing compound comprises an autophagy-inducing peptide comprising Beclin 1 residues 269-279 immediately N- and C-terminally flanked by moieties R 1 and R 2 , respectively, wherein up to six of said residues may be substituted, R 1 and R 2 do not naturally flank the Belclin 1 residues, and F270 and F274 are optionally substituted and optionally linked. The compounds may be used to induce autophagy.
1. An autophagy-inducing compound comprising an autophagy-inducing peptide comprising Beclin 1 residues 269-279 (SEQ ID NO:1) or the D-retro-inverso sequence thereof, immediately N- and C-terminally flanked by moieties R 1 and R 2 , respectively, wherein up to six of said residues may be substituted, R 1 and R 2 do not naturally flank the Beclin 1 residues, and F270 and F274 are optionally substituted and optionally linked.
2. The compound of claim 1 having the structure: R 1 -VFNATFEIWHD(SEQ ID NO:03)-R 2 , wherein up to six residues of SEQ ID NO:03 may be substituted, and the two F residues of SEQ ID NO:03 are optionally substituted and optionally linked to each other.
3. The compound of claim 1 wherein R 1 comprises:
a stabilizing agent selected from PEG, oligo-N-methoxyethylglycine (NMEG), albumin, an albumin-binding protein, and an immunoglobulin Fc domain, or
an affinity tag selected from an immuno-tag, biotin, lectin, and chelator, or
a label selected from an optical tag, a chelated lanthanide, a fluorescent dye, and a FRET acceptor/donor.
4. The compound of claim 1 , wherein R 1 comprises a transduction domain, a homing peptide, or a serum stabilizing agent.
5. The compound of claim 1 wherein R 1 is a tat protein transduction domain linked to the peptide through a diglycine linker, particularly a diglycine-T-N linker.
6. The compound of claim 1 wherein R 2 is carboxyl.
7. The compound of claim 1 wherein R 2 comprises an affinity tag or detectable label, particularly a fluorescent label.
8. The compound of claim 1 wherein F270 and F274 are substituted with crosslinkable moieties and/or linked, and each optionally comprises an additional α-carbon substitution selected from substituted, optionally hetero-lower alkyl, particularly optionally substituted, optionally hetero-methyl, ethyl, propyl and butyl.
9. The compound of claim 1 wherein F270 and F274 are substituted with homocysteines connected through a disulfide bridge to generate a ring and tail cyclic peptide.
10. The compound of claim 1 wherein the side chains of F270 and F274 are replaced by a linker:
(CH 2 ) n ONHCOX(CH 2 ) m —, wherein X is CH 2 , NH or O, and m and n are integers 1-4, forming a lactam peptide;
CH 2 OCH 2 CHCHCH 2 OCH 2 —, forming an ether peptide; or
(CH 2 ) n CHCH(CH 2 ) m —, forming a stapled peptide.
11. The compound of claim 1 wherein 1 to 6 residues are alanine substituted.
12. The compound of claim 1 , wherein the peptide comprises at least one of substitutions: H275E and S279D.
13. The compound of claim 1 , wherein the peptide comprises one or more D-amino acids, or one or more L-β-homo amino acids, or one or more D-β-homo amino acids, or one or more N-methylated amino acids.
14. The compound of claim 1 comprising the D-retro-inverso sequence.
15. The compound of claim 1 comprising the D-retro-inverso sequence: RRQRRKKKRGY-GG-DHWIEFTANFV (SEQ ID NO:08).
16. The compound of claim 1 wherein the peptide is acetylated, acylated, formylated, amidated, phosphorylated, sulfated or glycosylated.
17. The compound of claim 1 comprising an N-terminal acetyl, formyl, myristoyl, palmitoyl, carboxyl or 2-furosyl group, and/or a C-terminal hydroxyl, amide, ester or thioester group.
18. The compound of claim 1 wherein the peptide is cyclized.
19. A pharmaceutical composition comprising a compound of claim 1 in unit dosage, administrable form.
20. A method of inducing autophagy, comprising administering to a person in need thereof an effective amount of a compound of claim 1 .