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Title:  Method of inducing apoptosis in cancer cells using an extract of Melothria indica Lou

United States Patent:  6,312,737

Inventors:  Tsai; David (Westlake Village, CA)

Assignee:  Ambryx Biotechnology, Inc. ()

Appl. No.:  568625

Filed:  May 10, 2000

Abstract

The invention includes a method of inducing apoptosis in cancer cells by administering an extract of Melothria indica Lou to the cancer cells. The invention also includes a method of using an extract of Melothria indica Lou to induce apoptosis in cancer cells by administering the extract to the cancer cells. The cancer cells can be leukemia cells and prostate cancer cells. Further included in the invention is a method for purifying the Melothria indica Lou into an extract used for inducing apoptosis in cancer cells.

SUMMARY OF THE INVENTION

The present invention introduces such refinement. In its preferred embodiments, the present invention has several aspects or facets that can be used independently, although they are preferably employed together to optimize their benefits.

Melothria indica Lou (Melothria) is a perennial berbaceous vine that thrives on hillsides and in the woods and bushes. Melothria, which emerged recently, is a popular herb in Taiwan, China and India and is believed to be beneficial in the treatment of sore throats, acute conjunctivitis and inflammation.

We have discovered, however, that the Melothria extract selectively induces apoptosis in cancer cells.

Thus, the invention includes a method of inducing apoptosis in cancer cells by administering an extract of Melothria indica Lou to the cancer cells, with the greatest apoptotic effects known at this time occurring in leukemia cells and prostate cancer cells. The invention also includes a method of using an extract of Melothria indica Lou to induce apoptosis in cancer cells by administering the extract to the cancer cells.

The invention further includes a method for purifying Melothria indica Lou, which comprises the following steps:

Extracting the Melothria indica Lou with a solvent to form an extract, concentrating the extract, applying the extract to a SCD-100 reverse phase chromatography column equilibrated with phosphate buffer saline, developing the SCD-100 column using an isocratic elution of phosphate buffer saline for about forty (40) minutes with a flow rate of about 0.4 ml/min, collecting at least fraction eleven (11) from the column, applying fraction eleven to a RPP-100 reverse-phase chromatography column equilibrated with 0.1% trifluoroacetic acid in water, developing the column by a linear gradient using solvent A and solvent B, solvent A being 0.1% trifluoroacetic acid in water and solvent B being water/CH3 CN/0.1% trifluoroacetic acid in a ratio of 49.5:49.5:1, generating a gradient using 100% solvent A and 0% solvent B at time zero, then increasing solvent B from 0% to 100% after 20 ml elution volume, collecting at least fraction two from the column, applying fraction two to a AX-100 anion exchange chromatography column equilibrated with solvent C, solvent C being 10% phosphate buffer saline in water, eluting the column with a solvent gradient including solvent C and solvent D, solvent D being 100% phosphate buffer saline, the solvent gradient is created starting with 100% solvent C and 0% of solvent D at time zero, then increasing solvent D from 0% to 100% after 20 ml elution volume, collecting at least fraction 12 from the column, applying fraction twelve to a silica gel column equilibrated with a solvent E, solvent C being CH3 CN:ethanol:water at a ratio of 84.5:15:0.5, isocratically eluting the column with solvent C, collecting at least fractions eight and nine.

Claim 1 of 4 Claims

I claim:

1. A method of inducing apoptosis in leukemia cells by administering an extract of fruit of Melothria indica Lou to a subject in an effective amount to induce apoptosis of said leukemia.

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If you want to learn more about this patent, please go directly to the U.S. Patent and Trademark Office Web site to access the full patent.

 

 

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