Drug – bio-affecting and body treating compositions – Plant material or plant extract of undetermined constitution... – Containing or obtained from cucumbrits
Reexamination Certificate
2000-05-10
2001-11-06
Tate, Christopher R. (Department: 1651)
Drug, bio-affecting and body treating compositions
Plant material or plant extract of undetermined constitution...
Containing or obtained from cucumbrits
C424S725000, C514S001000, C514S908000
Reexamination Certificate
active
06312737
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates generally to cancer treatment and more specifically, to cancer treatments which selectively induce apoptosis in cancer cells.
2. Description of Related Art
Cancer was generally treated with combinations of surgery, chemotherapy and radiation with varying degrees of success. One way cancer has been targeted for treatment is that cancer cells tend to be rapidly dividing cells. Thus, current chemotherapeutic approaches target rapidly dividing tumor cells. This approach is generally ineffective when the cancer is dormant or slow growing. These types of treatments also impact other, noncancerous cells that divide rapidly, causing harmful side effects.
These harmful side effects, however, are not present in treatments that induce apoptosis in the cancer cells. Apoptosis is also called “programmed cell death” or “cell suicide.” (Krammer, et al., “Apoptosis in the APO-1 System”, Apoptosis: The molecular Basis of Cell Death, pp. 87-99 Cold Spring Harbor Laboratory Press, 1991). In contrast to the cell death caused by cell injury, apoptosis is an active process of gene-directed, cellular self-destruction. Apoptosis serves a biologically meaningful function. (Kerr, J. F. R. and J. Searle, J. Pathol. 107:41, 1971).
One of the examples of the biologically meaningful functions of apoptosis is the morphogenesis of an embryo. (Michaelson, J., Biol. Rev. 62:115, 1987). Just like creating a sculpture, clay is added, and clay is removed; organ formation (morphogenesis) of an embryo relies on cell growth (addition of clay) as well as cell death (removal of clay). As a matter of fact, apoptosis plays a key role in the human body from the early stages of embryonic development through to the inevitable decline associated with old age. (Wyllie, A. H., Int. Rev. Cytol. 68:251, 1980).
The normal function of the immune, gastrointestinal and hematopoietic system relies on the normal function of apoptosis. When the normal function of apoptosis goes awry, the result can be any of the following diseases: cancer, viral infections, autoimmune disease/allergies, neurodegeneration or cardiovascular diseases.
The idea that cancer may be caused by insufficient apoptosis emerged only recently (Cope, F. O. and Wille, J. J., “Apoptosis”: The Molecular Basis of Cell Death, Cold Spring Harbor Laboratory Press, p. 61, 1991). This idea opens a door for a new concept in cancer therapy—cancer cells may be killed by encouraging apoptosis. Apoptosis modulation, based on the processes present in normal development, is a potential mechanism for controlling the growth of tumor cells. Restoring apoptosis in tumor cells is an attractive approach because it programs the cancer cells to commit suicide.
Thus, the cancer cells can be killed without killing the host. The success of this treatment, however, is dependent upon the availability of drugs that can selectively induce apoptosis in tumor cells without affecting normal cells.
From the preceding descriptions, it is apparent that the compositions and methods currently being used have significant disadvantages. Thus, important aspects of the technology used in the field of invention remain amenable to useful refinement.
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/CH
3
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 CH
3
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.
All of the foregoing operational principles and advantages of the present invention will be more fully appreciated upon consideration of the following detailed description, with reference to the appended drawings.
REFERENCES:
patent: 40915938A (1997-12-01), None
patent: 10114670A (1998-05-01), None
Krammer, P.H., et. al., Apoptosis in the APO-1 System Apoptosis: The Molecular Basis of Cell Death, 1991, pp. 87-99, Cold Spring Harbor Lab. Press.
Kerr, J.F.R. and Searle, J., A Suggested Explanation for the Paradoxically slow growth rate of basal-cell Carcinomas . . . , J. Path., vol. 107, p. 41-44, (1971).
Michaelson, J., Cell Selection in Development, Biol. Rev. (1987), vol. 62, p.115-139, Great Britain.
Wyllie, A.H., et al., Cell Death: The Significance of Apoptosis, International Review of cytology, vol. 68, p. 251-306, Academic Press, 1980.
Cope, F., et al., Apoptosis: The Molecular Basis of Cell Death, Cold Spring Harbor Laboratory Press, p. 61-86, 1991: “Carciogenesis and Apoptosis: Paradigms . . . ”.
Ambryx Biotechnology, Inc.
Flood Michele
Tate Christopher R.
Trojan Law Offices
LandOfFree
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