Water soluble extract from plant ofSolanumgenus and the preparation process thereof, and pharmaceutical composition containing the water soluble extract

• US 7,078,063 B2
• Filed: 08/29/2003
• Issued: 07/18/2006
• Est. Priority Date: 06/05/2003
• Status: Active Grant

• Current Assignee: G & E Herbal Biotechnology Company Limited
• Original Assignee: G & E Herbal Biotechnology Company Limited
• Inventors: Kuo, Kou-Wha
• Primary Examiner(s): Coe, Susan D.
• Application Number:
  US10/650,942
• Publication Number:
  US 20040247715A1
• Time in Patent Office: 1,054 Days
• Field of Search: None
• US Class Current: 424/725
• CPC Class Codes: A61K 31/706 containing six-membered rings with nitrogen as a ring hetero atom
• CPC Class Codes: A61K 36/539 Scutellaria (skullcap)
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Abstract

A water soluble extract from a plant of Solanum genus consists essentially of at least 60%–90% of solamargine and solasonine. A process for preparing the water soluble extract from the plant of Solanum genus involves the steps of hydrolysis with an acid, precipitation with a base, and separation treatments using chloroform, alcohol and water as extraction solvents. The water soluble extract prepared from the process can be directly dissolved in pure or neutral pH water to form a yellowish clear and transparent aqueous solution having a water solubility ranging from 2˜20 mg/ml or higher.

The water soluble extract can be used as an active component in a pharmaceutical composition for inhibiting the growth of tumor/cancer cells, in particular liver cancer cells, lung cancer cells and breast cancer cells.

Claims

1. A water-soluble extract from a plant of Solanum genus, consisting essentially of at least 60%–

90% of solamargine and solasonine.

2. The water-soluble extract according to claim 1, which is extracted from a plant of Solanum genus selected from the group consisting of:

Solanum incanum L., Solanum indicum, Solanum nigrum, Solanum capsicastrum, Solanum xanthocarpum, Solanum melongena, Solanum coagulans, Solanum tunigrum, Solanum sodomeum, Solanum turburosum, Solanum aculeastrum, Solanum lycocarpum, Solanum khasianum, Solanum suaveolens, Solanum uporo, Solanum abutiloides, Solanum coccineum, Solanum unguiculatum, Solanum robustum, Solanum anguivi, Solanum platanifolium, Solanum mammosum, and a combination thereof.

3. The water-soluble extract according to claim 2, which is extracted from Solanum incanum L.

4. The water-soluble extract according to claim 2, which is extracted from Solanum nigrum.

5. The water-soluble extract according to claim 1, which is prepared from a process comprising the steps of:

(a) subjecting a plant material of a plant of Solanum genus to an extraction treatment using an acidic aqueous solution with a pH value of 3˜

5, such that an aqueous solution is obtained;

(b) adjusting the pH value of the aqueous solution obtained in step (a) to pH 8˜

10 with a base, such that a precipitate is formed;

(c) washing the precipitate formed in step (b) with water, followed by drying, such that a dried product is obtained;

(d) admixing the dried product obtained in step (c) with chloroform, followed by addition of a suitable amount of a 100% alcohol, such that a chloroform-alcohol mixture is formed;

(e) mixing the chloroform-alcohol mixture formed in step (d) with a water/alcohol solution having a predetermined water;

alcohol ratio, such that a mixture containing a chloroform-based layer and a non-chloroform-based layer is obtained;

(f) removing the chloroform-based layer from the mixture obtained in step (e), followed by addition of a suitable amount of water; and

(g) obtaining a supernatant from the resultant mixture of step (f), followed by drying the supernatant, wherein the resultant dried product is able to be directly dissolved in water to form a yellowish clear and transparent aqueous solution.

6. The water-soluble extract according to claim 5, wherein in step (a) of said process, the used plant material is at least one of the fruit, root, stem, and leaf of said plant of Solanum genus.

7. The water-soluble extract according to claim 6, wherein in step (a) of said process, the used plant material is the fruit of said plant of Solanum genus.

8. The water-soluble extract according to claim 6, wherein in step (a) of said process, the plant material is the whole plant of said plant of Solanum genus.

9. The water-soluble extract according to claim 5, wherein in step (a) of said process, the plant material of said plant of Solanum genus has been chopped in a preliminary treatment.

10. The water-soluble extract according to claim 5, wherein in step (a) of said process, the aqueous solution is obtained by conducting centrifugation subsequent to the extraction treatment.

11. The water-soluble extract according to claim 5, wherein in step (a) of said process, the acidic aqueous solution in the extraction treatment is an aqueous solution containing formic acid, acetic acid, or hydrochloric acid.

12. The water-soluble extract according to claim 5, wherein in step (b) of said process, the base is an alkaline aqueous solution containing a compound selected from the group consisting of alkali hydroxides and ammonium hydroxide.

13. The water-soluble extract according to claim 12, wherein in step (b) of said process, the base is an alkaline aqueous solution containing ammonium hydroxide.

14. The water-soluble extract according to claim 12, wherein in step (b) of said process, the base is an alkaline aqueous solution containing sodium hydroxide.

15. The water-soluble extract according to claim 5, wherein in step (b) of said process, the precipitate is obtained by conducting centrifugation subsequent to the pH value adjustment.

16. The water-soluble extract according to claim 5, wherein in step (c) of said process, the drying treatment is selected from the group consisting of lyophilization, spray-drying, evaporation, heat-drying, and a combination thereof.

17. The water-soluble extract according to claim 5, wherein, in step (c) of said process, the dried product is obtained by washing the precipitate formed in step (b) with water and suspending the washed precipitate in water, followed by lyophilization.

18. The water-soluble extract according to claim 5, wherein in steps (d) and (e) of said process, the alcohol is selected from the group consisting of methanol, ethanol, propyl alcohol, and a combination thereof.

19. The water-soluble extract according to claim 5, wherein in step (f) of said process, the removal of the chloroform-based layer is conducted by centrifugation.

20. The water-soluble extract according to claim 5, wherein in step (g) of said process, the drying treatment is selected from the group consisting of lyophilization, spray-drying, evaporation, heat-drying, and a combination thereof.

21. The water-soluble extract according to claim 1, which is in a form of water-soluble particles with a nanoparticle size.

22. The water-soluble extract according to claim 20, which is in a form of water-soluble particles with a particle size less than 1 μ

m.

23. The water-soluble extract according to claim 1, which is composed of more than 75% of solasonine and solamargine.

24. The water-soluble extract according to claim 1, which has a solasonine to solamargine ratio ranging from 0.3:

1.0 to 1.0;

0.6.

25. The water-soluble extract according to claim 1, which has a solasonine to solamargine ratio ranging from 0.4:

1.0 to 0.9;

1.0.

26. The water-soluble extract according to claim 1, which has a solasonine to solamargine ratio around 0.7:

1.0.

27. The water-soluble extract according to claim 1, which has a water solubility ranging from 2 to 20 mg/ml or higher.

28. The water-soluble extract according to any one of claims 1–

27, which can be used in manufacture of a medicament effective in inhibiting the growth of tumor/cancer cells.

29. A pharmaceutical composition, comprising a water-soluble extract according to any one of claims 1–

27 and, optionally, a pharmaceutically acceptable carrier.

30. A pharmaceutical composition for inhibiting the growth of tumor/cancer cells, comprising a water-soluble extract according to any one of claims 1–

27 and, optionally, a pharmaceutically acceptable carrier.

31. A process for preparing a water-soluble extract from a plant of Solanum genus, comprising the steps of:

(a) subjecting a plant material of a plant of Solanum genus to an extraction treatment using an acidic aqueous solution with a pH value of 3˜

5, such that an aqueous solution is obtained;

(b) adjusting the pH value of the aqueous solution obtained in step (a) to pH 8˜

10 with a base, such that a precipitate is formed;

(c) washing the precipitate formed in step (b) with water, followed by drying, such that a dried product is obtained;

(d) admixing the dried product obtained in step (c) with chloroform, followed by addition of a suitable amount of a 100% alcohol, such that a chloroform-alcohol mixture is formed;

(e) mixing the chloroform-alcohol mixture formed in step (d) with a water/alcohol solution having a predetermined water;

alcohol ratio, such that a mixture containing a chloroform-based layer and a non-chloroform-based layer is obtained;

(f) removing the chloroform-based layer from the mixture obtained in step (e), followed by addition of a suitable amount of water; and

(g) obtaining a supernatant from the resultant mixture of step (f), followed by drying the supernatant, wherein the resultant dried product is able to be directly dissolved in water to form a yellowish clear and transparent aqueous solution.

32. The process according to claim 31, wherein in step (a), the plant material of said plant of Solanum genus has been chopped in a preliminary treatment.

33. The process according to claim 31, wherein in step (a), the plant material is at least one of the fruit, root, stem, and leaf of said plant of Solanum genus.

34. The process according to claim 31, wherein in step (a), the plant material is the fruit of said plant of Solanum genus.

35. The process according to claim 31, wherein in step (a), the plant material is the whole plant of said plant of Solanum genus.

36. The process according to claim 31, wherein in step (a), the plant material is from a plant of Solanum genus selected from the group consisting of Solanum incanum L., Solanum indicum, Solanum nigrum, Solanum capsicastrum, Solanum xanthocarpum, Solanum melongena, Solanum coagulans, Solanum tunigrum, Solanum sodomeum, Solanum turburosum, Solanum aculeastrum, Solanum lycocarpum, Solanum khasianum, Solanum suaveolens, Solanum uporo, Solanum abutiloides, Solanum coccineum, Solanum unguiculatum, Solanum robustum, Solanum anguivi, Solanum platanifolium, Solanum mammosum, and a combination thereof.

37. The process according to claim 36, wherein in step (a), the plant material is from Solanum incanum L.

38. The process according to claim 36, wherein in step (a), the plant material is from Solanum nigrum.

39. The process according to claim 31, wherein in step (a), the aqueous solution is obtained by conducting centrifugation subsequent to the extraction treatment.

40. The process according to claim 31, wherein in step (a), the acidic aqueous solution in the extraction treatment is an aqueous solution containing formic acid, acetic acid, or hydrochloric acid.

41. The process according to claim 31, wherein in step (b), the base is an alkaline aqueous solution containing a compound selected from the group consisting of alkali hydroxides and ammonium hydroxide.

42. The process according to claim 41, wherein in step (b), the base is an alkaline aqueous solution containing ammonium hydroxide.

43. The process according to claim 41, wherein in step (b), the base is an alkaline aqueous solution containing sodium hydroxide.

44. The process according to claim 31, wherein in step (b), the precipitate is obtained by conducting centrifugation subsequent to the pH value adjustment.

45. The process according to claim 31, wherein in step (c), the drying treatment is selected from the group consisting of lyophilization, spray-drying, evaporation, heat-drying, and a combination thereof.

46. The process according to claim 31, wherein in step (c), the dried product is obtained by washing the precipitate formed in step (b) with water and suspending the washed precipitate in water, followed by lyophilization.

47. The process according to claim 31, wherein in steps (d) and (e), the alcohol is selected from the group consisting of methanol, ethanol, propyl alcohol, and a combination thereof.

48. The process according to claim 31, wherein, in step (f), the removal of the chloroform-based layer is conducted by centrifugation.

49. The process according to claim 31, wherein in step (g), the drying treatment is selected from the group consisting of:

lyophilization, spray-drying, evaporation, heat-drying, and a combination thereof.

50. The process according to claim 31, wherein the resultant product from step (g) is in a form of water-soluble particles with a nanoparticle size.

51. The process according to claim 50, wherein the resultant product from step (g) is in a form of water-soluble particles with a particle size less than 1 μ

m.

52. The process according to claim 31, wherein the resultant product from step (g) consists essentially of at least 60%–

90% of solasonine and solamargine.

53. The process according to claim 52, wherein the resultant product from step (g) is composed of more than 75% of solasonine and solamargine.

54. The process according to claim 31, wherein the resultant product from step (g) has a solasonine to solamargine ratio ranging from 0.3:

1.0 to 1.0;

0.6.

55. The process according to claim 31, wherein the resultant product from step (g) has a solasonine to solamargine ratio ranging from 0.4:

1.0 to 0.9;

1.0.

56. The process according to claim 31, wherein the resultant product from step (g) has a solasonine to solamargine ratio around 0.7:

1.0.

57. The process according to claim 31, wherein the resultant product from step (g) has a water solubility ranging from 2 to 20 mg/ml or higher.

58. A pharmaceutical composition comprising a water-soluble extract prepared by a process according to any one of claims 31–

57 and, optionally, a pharmaceutically acceptable carrier.

59. A pharmaceutical composition for inhibiting the growth of tumor/cancer cells, comprising a water-soluble extract prepared by a process according to any one of claims 31–

57 and, optionally, a pharmaceutically acceptable carrier.