Treating a melanoma with a whole, leech saliva extract
Methods are also provided for isolating and using a whole-saliva leech extract in the treatment of a melanoma. The methods can include feeding a phagostimulatory agent to a leech; inducing a regurgitation in the leech, the inducing including placing the leech in an environment having a temperature of less than about 0° C.; and, collecting an unrefined, whole saliva in the regurgitation of the cooled leech. The methods can include revitalizing the leech by warming it at a temperature ranging from about 5° C. to about 40° C. Stable, lyophilized, whole-saliva extracts of a leech are also provided, the extract having a stable activity when stored for use at a temperature below about −20° C., the extract maintaining at least 70% of the activity for at least 6 months. The extracts can be used to treat solid tumors, treat liquid tumors, treat diabetes, treat a viral disease, treat a parasitic disease, treat an antibacterial disease, or serve as an anti-oxidant.
1. A method of inhibiting the growth of melanoma cells, the method comprising
contacting a melanoma cell in a fluid with a whole, leech saliva extract; the extract having a peptide molecular weight distribution with molecular weights of 3496 Daltons or greater; wherein the extract is produced from a process comprising:
feeding a phagostimulatory agent to a leech;
inducing a regurgitation in the leech, the inducing including placing the leech in an environment having a temperature of less than 0° C. or about 0° C.; and, collecting an unrefined, whole saliva in the regurgitation of the cooled leech.
2. The method of claim 1 , further comprising revitalizing the leech by warming the leech in a water bath having a temperature ranging from about 5° C. to about 40° C.
3. The method of claim 2 , wherein the method further comprises creating a refined, whole-saliva extract; the creating including removing solid components from the unrefined, whole saliva.
4. The method of claim 2 , wherein the method further comprises lyophilizing separate volumes of the refined, whole saliva extract, the volumes not exceeding about 2 ml each.
5. The method of claim 1 , the leech being Hirudinaria manillensis.
6. A method of treating a melanoma in a subject, comprising administering an effective amount of a refined, whole leech saliva extract to the subject, the extract having a peptide molecular weight distribution with molecular weights of 3496 Daltons or greater; wherein the whole leech saliva extract functions as an agent in the treatment of the melanoma, and the extract is produced from a process comprising:
feeding a phagostimulatory agent to a leech;
inducing regurgitation in the leech, the inducing including placing the leech in an environment having a temperature ranging from about −5° C. to about 15° C.
collecting an unrefined, whole saliva in the regurgitation of the cooled leech;
removing solid components from the unrefined, whole saliva to create a refined, whole saliva; and,
lyophilizing separate volumes of the refined, whole saliva extract, the volumes not exceeding about 2 ml each.
7. The method of claim 6 , further comprising revitalizing the leech by warming the leech in a water bath having a temperature ranging from about 5° C. to about 40° C.
8. The method of claim 6 , the leech being Hirudinaria manillensis.
9. The method of claim 6 , wherein the administration includes administering a second agent selected from the group consisting of an immunomodulatory agent, an antiproliferative, an antineoplastic, an antimitotic, or an anti-inflammatory.
10. The method of claim 6 , wherein administration includes administering irinotecan.
11. The method of claim 6 , wherein administration includes administering carboplatin.
12. The method of claim 6 , wherein the administration includes administering a second agent selected from the group consisting of an antiplatelet, an anticoagulant, an antifibrin, an antithrombin, an antibiotic, an antiallergic, or an antioxidant.
13. The method of claim 6 , wherein the collecting of the whole saliva yields a peptide molecular weight distribution that ranges from 4276 Daltons to 44386 Daltons.
14. The method of claim 6 , wherein the collecting of the whole saliva yields a peptide molecular weight distribution that ranges from 6289 Daltons to 14244 Daltons.
15. The method of claim 6 , wherein the collecting of the whole saliva yields a peptide molecular weight distribution that ranges from 10812 Daltons to 88210 Daltons.
16. The method of claim 6 , wherein the collecting of the whole saliva yields a peptide molecular weight distribution that ranges from 3496 Daltons to 88210 Daltons.