Peptide toxin formulation
Procedures are described which use solvents to increase the topical insecticidal activity of toxic insect peptides. These procedures comprise drying the peptides, if needed, followed by the addition of either: 1) a polar organic solvent, with or without water, to a dried peptide, or 2) the addition of polar aprotic solvent or other adjuvant to the dried peptide, followed by the addition of either: 1) a polar organic solvent, with or without water, (where a polar aprotic solvent is added first) or 2) a polar aprotic solvent or other adjuvant to the peptide polar organic solvent (where the polar organic solvent is added first), to the peptide formulation.
1. A process for treating a toxic peptide to increase its topical insecticidal activity, comprising:
a) mixing a polar organic solvent with said peptide to make a polar organic solvent peptide suspension;
b) adding a polar aprotic solvent to said polar organic solvent peptide suspension to create a final peptide suspension;
wherein said final peptide suspension comprises 90% or more polar organic solvent and 10% or less polar aprotic solvent.
2. The process of claim 1 , wherein said polar aprotic solvent is selected from the group consisting of dimethyl sulfoxide, dimethylformamide, dioxane and hexamethylphosphorotriamide or any combination thereof.
3. The process of claim 2 , wherein said polar organic solvent is selected from the group consisting of acetone, methanol, ethanol, propanol, methyl ethyl ketone, diethyl ketone, acetonitrile, and ethyl acetoacetate or any combination thereof.
4. The process of claim 3 , wherein said polar aprotic solvent is selected from the group consisting of dimethyl sulfoxide and dimethylformamide or a combination of the two, and said polar organic solvent is selected from the group consisting of acetone, methanol, ethanol and propanol, or any combination thereof.
5. The process of claim 1 , wherein said toxic peptide is lyophilized before the first solvent is mixed with the toxic peptide.
6. The process of claim 1 , wherein either one or both of the peptide solvents are sonicated after mixing with said peptide.
7. The process of claim 1 , wherein said polar organic solvent and said polar aprotic solvent form a total solvent volume of 100%, and wherein said polar aprotic solvent—comprises from about 8% to about 0.1% of the total solvent volume, and said polar organic solvent comprises from about 92% to about 99.9% of the total solvent volume.
8. The process of claim 7 , wherein said polar aprotic solvent comprises from about 5% to about 0.1% of the total solvent volume, and the polar organic solvent comprises from about 95% to about 99.9% of the total solvent volume.
9. The process of claim 7 , wherein said polar aprotic solvent comprises from about 2% to about 0.1% of the total solvent volume, and the polar organic solvent comprises from about 98% to about 99.9% of the total solvent volume.
10. The process of claim 1 , wherein said toxic peptide has at least 50% sequence identity to a natural peptide produced by a spider, scorpion, snake, mite, snail, or plant.
11. The process of claim 10 , wherein said peptide is produced by a spider.
12. The process of claim 11 , wherein said peptide is produced by the Australian funnel web spider of genus Atrax or Hadronyche.
13. A toxic peptide suspension produced by the process of claim 10 .
14. The process of claim 1 , wherein the length of said toxic peptide is from about 10 to 200 amino acids.
15. The process of claim 1 , wherein said toxic peptide has from 1 to 5 disulfide bonds.
16. The process of claim 1 , wherein said toxic peptide comprises a peptide sequence with at least 50% identity to any one of SEQ ID NOs: 1 to 174.
17. The process of claim 16 , wherein said peptide sequence has at least 70% identity to any one of SEQ ID NOs: 1 to 174.
18. The process of claim 16 , wherein said peptide sequence is selected from the group consisting of SEQ ID NO: 60, SEQ ID NO: 117, SEQ ID NO: 118, and SEQ ID NO: 119.
19. The process of claim 16 , wherein said peptide sequence has at least 85% identity to any one of SEQ ID NOs: 1 to 174.
20. The process of claim 16 , wherein said peptide sequence has at least 90% identity to any one of SEQ ID NOs: 1 to 174.
21. A toxic peptide suspension produced by the process of claim 1 .
22. A peptide formulation comprising:
a) an insecticidal peptide;
b) a polar organic solvent;
wherein said polar organic solvent comprises from about 90% to about 99% of the final solvent volume of the formulation; and
c) a polar aprotic solvent; optionally containing water;
wherein said polar aprotic solvent comprises from about 10% to about 1% of the final solvent volume of the formulation; and
wherein said optional water phase comprises from 0% to about 10% of the final solvent volume of the formulation.