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 method of controlling an insect comprising applying a peptide suspension to the insect's environment, wherein said peptide suspension is made by a process for treating a toxic peptide to increase its topical insecticidal activity, said process comprising:
a) mixing a polar organic solvent with said toxic 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, of at least or about 1.5 mg/ml wherein said final peptide suspension comprises 90% or more polar organic solvent and 10% or less polar aprotic solvent.
2. The method 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 method 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 method 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 method of claim 1 , wherein said toxic peptide is lyophilized before the first solvent is mixed with the toxic peptide.
6. The method of claim 1 , wherein either one or both of the peptide solvents are sonicated after mixing with said peptide.
7. The method 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 method of claim 7 , wherein said polar aprotic solvent [H] 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 method of claim 8 , wherein said polar aprotic solvent [H] 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 method 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 method of claim 10 , wherein said toxic peptide is produced by a spider.
12. The method of claim 11 , wherein said toxic peptide is produced by the Australian funnel web spider of genus Atrax or Hadronyche.
13. The method of claim 1 , wherein the length of said toxic peptide is from about 10 to 200 amino acids.
14. The method of claim 1 , wherein said toxic peptide has from 1 to 5 disulfide bonds.
15. The method of claim 1 , wherein said toxic peptide has at least 50% sequence identity to any one of SEQ ID NOs: 1 to 174.
16. The method of claim 15 , wherein said toxic peptide has at least 70% sequence identity to any one of SEQ ID NOs: 1 to 174.
17. The method of claim 16 , wherein said toxic 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.
18. The method of claim 16 wherein said toxic peptide has at least 85% sequence identity to any one of SEQ ID NOs: 1 to 174.
19. The method of claim 18 wherein said toxic peptide has at least 90% sequence identity to any one of SEQ ID NOs: 1 to 174.
20. A method of controlling an insect comprising applying a peptide formulation of at least or about 1.5 mg/ml to the insect's environment, wherein said peptide formulation is comprised of:
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.
21. The method of claim 20 wherein said insecticidal peptide has at least 50% sequence identity to a natural peptide produced by a spider, scorpion, snake, mite, snail, or plant.