Human cathelicidin antimicrobial peptides
View Patent ↗Provided are peptide and peptide consensus sequences, which inhibit bacterial growth and/or viral growth and mimic the activity of LL-37, CRAMP, and/or FALL-39. The peptides are useful as antimicrobials, anti-inflammatories and antiviral agents.
1. A substantially purified polypeptide:
(a) consisting of 16-20 amino acids in length; and
(b) containing the sequence X 1 X 2 X 3 X 4 X 5 X 6 IKX 7 FX 8 X 9 X 10 LX 11 P (SEQ ID NO:1), wherein X 1 , X 2 , and X 6 are individually K or R; wherein X 3 is I or K; wherein X 4 is V or G; wherein X 5 is Q or R; wherein X 7 , X 9 , X 10 , and X 11 are each individually any amino acid; wherein X 8 is L or F and wherein the polypeptide comprises antibacterial and/or, antifungal activity.
2. The substantially purified polypeptide of claim 1 , wherein the polypeptide consists of a sequence selected from the sequence consisting of:
(a)
KRIVQRIKDFLRNLVP;
(SEQ ID NO: 13)
(b)
KRIVQRIKDFLRNLVPR;
(SEQ ID NO: 14)
(c)
KRIVQRIKDFLRNLVPRT;
(SEQ ID NO: 15)
(d)
KRIVQRIKDFLRNLVPRTE;
(SEQ ID NO: 16)
and
(e)
KRIVQRIKDFLRNLVPRTES.
(SEQ ID NO: 17)
3. A substantially purified polypeptide
(a) consisting of about 26 to 28 amino acids in length; and
(b) containing the sequence X 1 X 2 X 3 X 4 X 5 X 6 IKX 7 FX 8 X 9 X 10 LX 11 P (SEQ ID NO:1), wherein X 1 , X 2 , and X 6 are individually K or R; wherein X 3 is I or K; wherein X 4 is V or G; wherein X 5 is Q or R; wherein X 7 , X 9 , X 10 , and X 11 are each individually any amino acid; wherein X 8 is L or F and wherein the polypeptide comprises antibacterial and/or, antifungal activity.
4. The substantially purified polypeptide of claim 3 , wherein the polypeptide comprises a sequence selected from the group consisting of:
(SEQ ID NO: 18)
(a)
KSKEKIGKEFKRIVQRIKDFLRNLVP;
(SEQ ID NO: 19)
(b)
KSKEKIGKEFKRIVQRIKDFLRNLVPR;
(SEQ ID NO: 20)
(c)
KSKEKIGKEFKRIVQRIKDFLRNLVPRT;
(SEQ ID NO: 21)
(d)
KSKEKIGKEFKRIVQRIKDFLRNLVPRTE;
and
(SEQ ID NO: 22)
(e)
KSKEKIGKEFKRIVQRIKDFLRNLVPRTES.
5. A substantially purified polypeptide consisting of a sequence selected from the group consisting of:
(SEQ ID NO: 23)
(a)
RKSKEKIGKEFKRIVQRIKDFLRNLVP;
(SEQ ID NO: 24)
(b)
RKSKEKIGKEFKRIVQRIKDFLRNLVPR
(SEQ ID NO: 25)
(c)
RKSKEKIGKEFKRIVQRIKDFLRNLVPRT;
(SEQ ID NO: 26)
(d)
RKSKEKIGKEFKRIVQRIKDFLRNLVPRTE;
(SEQ ID NO: 27)
(e)
RKSKEKIGKEFKRIVQRIKDFLRNLVPRTES
(SEQ ID NO: 28)
(f)
LGDFFRKSKEKIGKEFKRIVQRIKDELRNLVPRTES.
6. A method of inhibiting the growth of a bacteria or fungus comprising contacting the bacteria or fungus with an inhibiting effective amount of a peptide
(a) consisting of 16-20 amino acids in length; and
(b) containing the sequence X 1 X 2 X 3 X 4 X 5 X 6 IKX 7 FX 8 X 9 X 10 LX 11 P (SEQ ID NO:1), wherein X 1 , X 2 , and X 6 are individually K or R; wherein X 3 is I or K; wherein X 4 is V or G; wherein X 5 is Q or R; wherein X 7 , X 9 , X 10 , and X 11 are each individually any amino acid; wherein X 8 is L or F and wherein the polypeptide comprises antibacterial and/or antifungal activity.
7. The method of claim 6 , wherein the peptide consists of a sequence selected from the group consisting of:
(a)
KRIVQRIKDFLRNLVP;
(SEQ ID NO: 13)
(b)
KRIVQRIKDFLRNLVPR;
(SEQ ID NO: 14)
(c)
KRIVQRIKDFLRNLVPRT;
(SEQ ID NO: 15)
(d)
KRIVQRIKDFLRNLVPRTE;
(SEQ ID NO: 16)
and
(e)
KRIVQRIKDFLRNLVPRTES.
(SEQ ID NO: 17)
8. A method of inhibiting the growth of a bacteria or fungus comprising contacting the bacteria or fungus with an inhibiting effective amount of a peptide
(a) consisting of about 26 to 28 amino acids in length; and
(b) containing the sequence X 1 X 2 X 3 X 4 X 5 X 6 IKX 7 FX 8 X 9 X 10 LX 11 P (SEQ ID NO:1), wherein X 1 , X 2 , and X 6 are individually K or R; wherein X 3 is I or K; wherein X 1 is V or G; wherein X 5 is Q or R; wherein X 7 , X 9 , X 10 , and X 11 are each individually any amino acid; wherein X 8 is L or F and wherein the polypeptide comprises antibacterial and/or antifungal activity.
9. The method of claim 8 , wherein the peptide comprises a sequence selected from the group consisting of:
(SEQ ID NO: 18)
(a)
KSKEKIGKEFKRIVQRIKDFLRNLVP;
(SEQ ID NO: 19)
(b)
KSKEKIGKEFKRIVQRIKDFLRNLVPR;
(SEQ ID NO: 20)
(c)
KSKEKIGKEFKRIVQRIKDFLRNLVPRT;
(SEQ ID NO: 21)
(d)
KSKEKIGKEFKRIVQRIKDFLRNLVPRTE;
and
(SEQ ID NO: 22)
(e)
KSKEKIGKEFKRIVQRIKDFLRNLVPRTES.
10. A method of inhibiting the growth of a bacteria or fungus comprising contacting the bacteria or fungus with an inhibiting effective amount of a polypeptide consisting of a sequence selected from the group consisting of:
(SEQ ID NO: 23)
(a)
RKSKEKIGKEFKRIVQRIKDFLRNLVP;
(SEQ ID NO: 24)
(b)
RKSKEKIGKEFKRIVQRIKDFLRNLVPR;
(SEQ ID NO: 25)
(c)
RKSKEKIGKEFKRIVQRIKDFLRNLVPRT;
(SEQ ID NO: 26)
(d)
RKSKEKIGKEFKRIVQRIKDFLRNLVPRTE;
(SEQ ID NO: 27)
(e)
RKSKEKIGKEFKRIVQRIKDFLRNLVPRTES.
11. The method of claim 6 , 8 , or 10 , wherein the contacting is in vitro.
12. The method of claim 11 , wherein the contacting is on a surface suspected of having a bacteria or fungus.
13. The method of claim 6 , 8 or 10 , wherein the contacting is in vivo.
14. The method of claim 13 , wherein the contacting in vivo is by topical administration.
15. The method of claim 6 , 8 or 10 , wherein the bacteria is gram positive.
16. The method of claim 15 , wherein bacteria is Staphylococcus aureus or S. epidermidis.
17. The method of claim 6 , 8 or 10 , wherein the bacteria is gram negative.
18. The method of claim 17 , wherein the bacteria is selected from the group consisting of E. coli, P. aeruginosa , and S. typhimurium.
19. The method of claim 6 , 8 or 10 , wherein the peptide is administered in combination with at least one antibiotic.
20. The method of claim 19 , wherein the class of antibiotic is selected from the group consisting of aminoglycosides, penicillins, cephalosporins, carbapenems, monobactams, quinolones, tetracyclines, glycopeptides, chloramphenicol, clindamycin, trimethoprim, sulfamethoxazole, nitrofurantoin, rifampin and mupirocin.
21. The method of claim 19 , wherein the antibiotic is selected from the group consisting of amikacin, gentamicin, kanamycin, netilmicin, tobramycin, streptomycin, azithromycin, clarithromycin, erythromycin, erythromycin estolate/ethylsuccinate/gluceptatellactobionate/stearate, penicillin G, penicillin V, methicillin, nafcillin, oxacillin, cloxacillin, dicloxacillin, ampicillin, amoxicillin, ticarcillin, carbenicillin, mezlocillin, azlocillin, piperacillin, cephalothin, cefazolin, cefaclor, cefamandole, cefoxitin, cefuiroxime, cefonicid, cefmetazole, cefotetan, cefprozil, loracarbef, cefetamet, cefoperazone, cefotaxime, ceftizoxime, ceftriaxone, ceftazidime, cefepime, cefixime, cefpodoxime, cefsulodin, imipenem, aztreonam, fleroxacin, nalidixic acid, norfloxacin, ciprofloxacin, ofloxacin, enoxacin, lomefloxacin, cinoxacin, doxycycline, minocycline, tetracycline, vancomycin, and teicoplanin.
22. A composition comprising a substantially purified polypeptide of claim 1 and a pharmaceutically acceptable carrier.
23. The composition of claim 22 , wherein the composition is a lotion, cream, gel, ointment or spray.
24. The pharmaceutical composition of claim 22 , wherein the pharmaceutical compositions is for topical administration.
25. The pharmaceutical composition of claim 22 , comprising a carbonate buffer system.
26. The pharmaceutical composition of claim 25 , wherein the carbonate buffer system comprises a bicarbonate concentration of between about 15 mM and about 500 mM bicarbonate.
27. A method of decontaminating a surface comprising contacting the surface with a composition comprising a cathelicidin functional fragment
(a) consisting of 16-20 amino acids in length; and
(b) containing the sequence X 1 X 2 X 3 X 4 X 5 X 6 IKX 7 FX 8 X 9 X 10 LX 11 P (SEQ ID NO:1), wherein X 1 , X 2 , and X 6 are individually K or R; wherein X 3 is I or K; wherein X 1 is V or G; wherein X 5 is Q or R; wherein X 7 , X 9 , X 10 , and X 11 are each individually any amino acid; wherein X 8 is L or F and wherein the polypeptide comprises antibacterial and/or antifungal activity.
28. The method of claim 27 , wherein the composition comprises a carbonate buffer.
29. The method of claim 28 , wherein the carbonate buffer comprises bicarbonate.
30. The method of claim 29 , wherein the bicarbonate comprises between about 15 mM and 500 mM bicarbonate.
31. The method of claim 27 , wherein the composition is in the form of a lotion, cream, gel, ointment or spray.
32. The method of claim 27 , wherein the polypeptide comprises a sequence selected from the sequence consisting of:
(a)
KRIVQRIKDFLRNLVP;
(SEQ ID NO: 13)
(b)
KRIVQRIKDFLRNLVPR-;
(SEQ ID NO: 14)
(c)
KRIVQRIKDFLRNLVPRT;
(SEQ ID NO: 15)
(d)
KRIVQRIKDFLRNLVPRTE;
(SEQ ID NO: 16)
and
(e)
KRIVQRIKDFLRNLVPRTES.
(SEQ ID NO: 17)
33. A method of decontaminating a surface comprising contacting the surface with a composition comprising a cathelicidin functional fragment that
(a) is about 26 to 28 amino acids in length; and
(b) contains the sequence X 1 X 2 X 3 X 4 X 5 X 6 IKX 7 FX 8 X 9 X 10 LX 11 P (SEQ ID NO:1), wherein X 1 , X 2 , and are individually K or R; wherein X 3 is I or K; wherein X 1 is V or G; wherein X 5 is Q or R; wherein X 7 , X 9 , X 10 , and X 11 are each individually any amino acid; wherein X 8 is L or F and wherein the polypeptide comprises antibacterial and/or antifungal activity.
34. The method of claim 33 , wherein the polypeptide comprises a sequence selected from the group consisting of:
(SEQ ID NO: 18)
(a)
KSKEKIGKEFKRIVQRIKDFLRNLVP;
(SEQ ID NO: 19)
(b)
KSKEKIGKEFKRIVQRIKDFLRNLVPR;
(SEQ ID NO: 20)
(c)
KSKEKIGKEFKRIVQRIKDFLRNLVPRT;
(SEQ ID NO: 21)
(d)
KSKEKIGKEFKRIVQRIKDFLRNLVPRTE;
and
(SEQ ID NO: 22)
(e)
KSKEKIGKEFKRIVQRIKDFLRNLVPRTES.
35. A method of decontaminating a surface comprising contacting the surface with a composition comprising a cathelicidin functional fragment consists of a sequence selected from the group consisting of:
(SEQ ID NO: 23)
(a)
RKSKEKIGKEFKRIVQRIKDFLRNLVP;
(SEQ ID NO: 24)
(b)
RKSKEKIGKEFKRIVQRIKDFLRNLVPR;
(SEQ ID NO: 25)
(c)
RKSKEKIGKEFKRIVQRIKDFLRNLVPRT;
(SEQ ID NO: 26)
(d)
RKSKEKIGKEFKRIVQRIKDFLRNLVPRTE;
(SEQ ID NO: 27)
(e)
RKSKEKIGKEFKRIVQRIKDFLRNLVPRTES
(SE ID NO: 28)
(f)
LGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES.