Chlorotoxin variants, conjugates, and methods for their use
Chlorotoxin variants, chlorotoxin variant conjugates, compositions that include the chlorotoxin variants or conjugates, and methods for using the chlorotoxin variants, conjugates, and compositions.
1. A chlorotoxin conjugate comprising a modified chlorotoxin polypeptide covalently conjugated to a fluorescent moiety comprising indocyanine green, the modified chlorotoxin polypeptide comprising an amino acid sequence corresponding to that of native chlorotoxin (SEQ ID NO: 1) in which lysine residue K15, lysine residue K23, or a combination thereof are substituted with an amino acid other than lysine, and wherein the chlorotoxin conjugate binds to chlorotoxin binding sites expressed by cancer cells.
2. The chlorotoxin conjugate of claim 1 , wherein the lysine residue K15 and the lysine residue K23 of the modified chlorotoxin polypeptide are substituted by an amino acid other than lysine.
3. The chlorotoxin conjugate of claim 2 , wherein the amino acid other than lysine is a natural amino acid or a non-natural amino acid.
4. The chlorotoxin conjugate of claim 2 , wherein the amino acid other than lysine is alanine or arginine.
5. The chlorotoxin conjugate of claim 1 , wherein the modified chlorotoxin polypeptide comprises a lysine residue at position 27.
6. The chlorotoxin conjugate of claim 1 , wherein the modified chlorotoxin polypeptide comprises a lysine residue at position 15.
7. The chlorotoxin conjugate of claim 1 , wherein the modified chlorotoxin polypeptide comprises a lysine residue at position 23.
8. The chlorotoxin conjugate of claim 1 , wherein the modified chlorotoxin polypeptide is conjugated to a therapeutic, diagnostic, imaging, or targeting agent, or a moiety that increases the circulatory half-life of the modified chlorotoxin polypeptide.
9. The chlorotoxin conjugate of claim 8 , wherein the therapeutic agent is a chemotherapeutic agent or a biological therapeutic agent.
10. The chlorotoxin conjugate of claim 8 , wherein the therapeutic agent is methotrexate, docetaxel, cisplatin, etoposide, cDNA, siRNA, shRNA, or RNAi.
11. The chlorotoxin conjugate of claim 8 , wherein the diagnostic or imaging agent is a fluorescent label, a radiolabel, or a magnetic resonance imaging label.
12. The chlorotoxin conjugate of claim 1 , wherein the modified chlorotoxin polypeptide comprises the amino acid sequence of SEQ ID NO: 1 in which the lysine residue K15, the lysine residue K23, or a combination thereof are substituted with a basic, non-polar, polar or acidic amino acid, or a non-natural amino acid, amino acid analog, or amino acid mimetic thereof.
13. The chlorotoxin conjugate of claim 1 , wherein the modified chlorotoxin polypeptide comprises the amino acid sequence of SEQ ID NO: 1 in which the lysine residue K15 and the lysine residue K23 are substituted by alanine or arginine.
14. The chlorotoxin conjugate of claim 1 , wherein the modified chlorotoxin polypeptide comprises the amino acid sequence of SEQ ID NO: 1 in which the lysine residue K15 is substituted by alanine and the lysine residue K23 is substituted by arginine.
15. The chlorotoxin conjugate of claim 1 , wherein the modified chlorotoxin polypeptide comprises the amino acid sequence of SEQ ID NO: 1 in which the lysine residue K15 is substituted by arginine and the lysine residue K23 is substituted by alanine.
16. A tissue or cell that has been removed from a subject, the tissue or cell comprising a cancerous tissue or cancerous cell bound to the chlorotoxin conjugate of claim 1 .
17. The chlorotoxin conjugate of claim 1 , wherein the modified chlorotoxin polypeptide has an amino acid sequence selected from the group consisting of:
SEQ ID NO: 2;
SEQ ID NO: 3;
SEQ ID NO: 4;
SEQ ID NO: 5; and
SEQ ID NO: 6.
18. The chlorotoxin conjugate of claim 6 , wherein the indocyanine green is conjugated to the lysine residue at position 15.
19. The chlorotoxin conjugate of claim 7 , wherein the indocyanine green is conjugated to the lysine residue at position 23.
20. The chlorotoxin conjugate of claim 5 , wherein the indocyanine green is conjugated to the lysine residue at position 27.
21. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a chlorotoxin conjugate comprising a modified chlorotoxin polypeptide covalently conjugated to a fluorescent moiety comprising indocyanine green, the modified chlorotoxin polypeptide comprising an amino acid sequence corresponding to that of native chlorotoxin (SEQ ID NO: 1) in which lysine residue K15, lysine residue K23, or a combination thereof are substituted with an amino acid other than lysine, and wherein the chlorotoxin conjugate binds to chlorotoxin binding sites expressed by cancer cells.
22. The pharmaceutical composition of claim 21 , wherein the modified chlorotoxin polypeptide comprises a lysine residue at position 27.
23. The pharmaceutical composition of claim 21 , wherein the modified chlorotoxin polypeptide has an amino acid sequence selected from the group consisting of:
SEQ ID NO: 2;
SEQ ID NO: 3;
SEQ ID NO: 4;
SEQ ID NO: 5; and
SEQ ID NO: 6.