Proteinaceous molecules and methods of use
This invention relates to peptides and their use for modulating sodium channels. More particularly, the present invention relates to peptides and their use in methods of enhancing Na v 1.1 activity and for treating or preventing conditions associated with Na v 1.1 activity.
1. A method of enhancing Na v 1.1 activity, comprising contacting a Na v 1.1 expressing cell with an isolated, synthetic or recombinant peptide comprising, consisting or consisting essentially of SEQ ID NO: 1:
[SEQ ID NO: 1]
Xaa 1 CRYLFGGCXaa 2 Xaa 3 TXaa 4 DCCKHLXaa 5 CRXaa 6 DXaa 7 Xaa 8 YCZ 1
wherein:
Xaa 1 is absent or is selected from acidic amino acid residues, including Asp and Glu;
Xaa 2 is selected from small amino acid residues, including Ser and Thr, and basic amino acid residues, including Arg, His and Lys;
Xaa 3 is selected from selected from small amino acid residues, including Ser and Thr;
Xaa 4 is selected from small amino acid residues, including Ser, Thr, Ala and Gly;
Xaa 5 is selected from small amino acid residues, including Ser, Thr, Ala and Gly;
Xaa 6 is selected from small amino acid residues, including Ser and Thr;
Xaa 7 is selected from aromatic amino acid residues, including Phe, Trp and Tyr, and hydrophobic amino acid residues, including Ile, Leu and Val;
Xaa 8 is selected from aromatic amino acid residues, including Phe, Trp and Tyr, and basic amino acid residues, including Arg, His and Lys; and
Z 1 is absent or is an amino acid sequence of SEQ ID NO: 2:
[SEQ ID NO: 2]
AWDGTFXaa 9
wherein:
Xaa 9 is absent or is a small amino acid residue, including Ser and Thr;
wherein the peptide is other than a peptide consisting of the amino acid sequence of SEQ ID NO: 4:
[SEQ ID NO: 4]
ECRYLFGGCSSTSDCCKHLSCRSDWKYCAWDGTFS.
2. A method of treating or preventing a condition in respect of which enhancing Na v 1.1 activity is associated with effective treatment, comprising administration of an isolated, synthetic or recombinant peptide comprising, consisting or consisting essentially of SEQ ID NO: 1:
[SEQ ID NO: 1]
Xaa 1 CRYLFGGCXaa 2 Xaa 3 TXaa 4 DCCKHLXaa 5 CRXaa 6 DXaa 7 Xaa 8 YCZ 1
wherein:
Xaa 1 is absent or is selected from acidic amino acid residues, including Asp and Glu;
Xaa 2 is selected from small amino acid residues, including Ser and Thr, and basic amino acid residues, including Arg, His and Lys;
Xaa 3 is selected from selected from small amino acid residues, including Ser and Thr;
Xaa 4 is selected from small amino acid residues, including Ser, Thr, Ala and Gly;
Xaa 5 is selected from small amino acid residues, including Ser, Thr, Ala and Gly;
Xaa 6 is selected from small amino acid residues, including Ser and Thr;
Xaa 7 is selected from aromatic amino acid residues, including Phe, Trp and Tyr, and hydrophobic amino acid residues, including Ile, Leu and Val;
Xaa 8 is selected from aromatic amino acid residues, including Phe, Trp and Tyr, and basic amino acid residues, including Arg, His and Lys; and
Z 1 is absent or is an amino acid sequence of SEQ ID NO: 2:
[SEQ ID NO: 2]
AWDGTFXaa 9
wherein:
Xaa 9 is absent or is a small amino acid residue, including Ser and Thr.
3. The method according to claim 2 , wherein the condition is epilepsy or Alzheimer's disease.
4. The method according to claim 3 , wherein the epilepsy is selected from Dravet syndrome, generalised epilepsy with febrile seizures plus, borderline severe myoclonic epilepsy of infancy and intractable childhood epilepsy with generalised tonic-clonic seizures.
5. The method according to claim 4 , wherein the epilepsy is Dravet syndrome.
6. The method according to claim 2 , wherein Xaa 1 is Glu.
7. The method according to claim 2 , wherein Xaa 2 is Lys or Ser.
8. The method according to claim 2 , wherein Xaa 4 is Ser or Ala.
9. The method according to claim 2 , wherein Xaa 5 is Gly or Ser.
10. The method according to claim 2 , wherein Xaa 7 is Leu or Trp.
11. The method according to claim 2 , wherein Xaa 8 is Lys or Tyr.
12. The method according to claim 2 , wherein Z 1 is the amino acid sequence of SEQ ID NO: 2.
13. The method according to claim 2 , wherein Xaa 9 is Ser.
14. The method according to claim 2 , wherein the peptide of SEQ ID NO: 1 comprises, consists or consists essentially of the amino acid sequence of SEQ ID NO: 4 or 5:
[SEQ ID NO: 4]
ECRYLFGGCSSTSDCCKHLSCRSDWKYCAWDGTFS
or
[SEQ ID NO: 5]
ECRYLFGGCKTTADCCKHLGCRTDLYYCAWDGTF.
15. The method according to claim 2 , wherein the six cysteine residues in the peptide are bonded in pairs to form three disulfide bonds.
16. The method according to claim 15 , wherein the disulfide bonds are formed between the side chains of Cys 2 and Cys 16, Cys 9 and Cys 21, and Cys 15 and Cys 28.
17. An isolated, synthetic or recombinant peptide comprising,
consisting or consisting essentially of SEQ ID NO: 1:
[SEQ ID NO: 1]
Xaa 1 CRYLFGGCXaa 2 Xaa 3 TXaa 4 DCCKHLXaa 5 CRXaa 6 DXaa 7 Xaa 8 YCZ 1
wherein
Xaa 1 is absent or is selected from acidic amino acid residues, including Asp and Glu;
Xaa 2 is selected from small amino acid residues, including Ser and Thr, and basic amino acid residues, including Arg, His and Lys;
Xaa 3 is selected from selected from small amino acid residues, including Ser and Thr;
Xaa 4 is selected from small amino acid residues, including Ser, Thr, Ala and Gly;
Xaa 5 is selected from small amino acid residues, including Ser, Thr, Ala and Gly;
Xaa 6 is selected from small amino acid residues, including Ser and Thr;
Xaa 7 is selected from aromatic amino acid residues, including Phe, Trp and Tyr, and hydrophobic amino acid residues, including Ile, Leu and Val;
Xaa 8 is selected from aromatic amino acid residues, including Phe, Trp and Tyr, and basic amino acid residues, including Arg, His and Lys;
Z 1 is absent or is an amino acid sequence of SEQ ID NO: 2:
[SEQ ID NO: 2]
AWDGTFXaa 9
wherein:
Xaa 9 is absent or is a small amino acid residue, including Ser and Thr; and
wherein the peptide is other than a peptide consisting of the amino acid sequence of
SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 10:
[SEQ ID NO: 4]
ECRYLFGGCSSTSDCCKHLSCRSDWKYCAWDGTFS
[SEQ ID NO: 5]
ECRYLFGGCKTTADCCKHLGCRTDLYYCAWDGTF
[SEQ ID NO: 10]
ECRYLFGGCKTTADCCKHLGCRTDLYYCAWDGTF-NH 2.
18. The peptide according to claim 17 , wherein Xaa 1 is Glu.
19. The peptide according to claim 17 , wherein Xaa 2 is Lys or Ser.
20. The peptide according to claim 17 , wherein Xaa 4 is Ser or Ala.
21. The peptide according to claim 17 , wherein Xaa 5 is Gly or Ser.
22. The peptide according to claim 17 , wherein Xaa 7 is Leu or Trp.
23. The peptide according to claim 17 , wherein Xaa 8 is Lys or Tyr.
24. The peptide according to claim 17 , wherein Z a is the amino acid sequence of SEQ ID NO: 2.
25. The peptide according to claim 17 , wherein Xaa 9 is absent.
26. The peptide according to claim 17 , wherein the peptide comprises, consists or consists essentially of the amino acid sequence of SEQ ID NO: 8:
[SEQ ID NO: 8]
SECRYLFGGCKTTADCCKHLGCRTDLYYCAWDGTF.
27. The peptide according to claim 17 , wherein the six cysteine residues are bonded in pairs to form three disulfide bonds.
28. The peptide according to claim 27 , wherein the disulfide bonds are formed between the side chains of Cys 2 and Cys 16, Cys 9 and Cys 21, and Cys 15 and Cys 28.
29. A composition comprising a peptide according to claim 17 and a pharmaceutically acceptable carrier or diluent.