Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Ketone doai
Reexamination Certificate
1999-05-28
2001-04-17
Goldberg, Jerome D. (Department: 1614)
Drug, bio-affecting and body treating compositions
Designated organic active ingredient containing
Ketone doai
C514S691000
Reexamination Certificate
active
06218434
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to the use of
Nigella sativa
derivatives in the treatment of carcinoma. More specifically, the invention relates to the use of the derivatives thymoquinone (TQ) and dithymoquinone (DIM) in the treatment of parental and multi-drug resistant (MDR) human cancers.
BACKGROUND OF THE INVENTION
Various anti-cancer drugs (a.k.a., antineoplastic agents) are widely used for treatment of both tumorous and leukemic parental cancer cells. Certain cell lines, however, are multi-drug resistant (MDR) and many standard anti-cancer drugs are MDR substrates. MDR, a phenomenon characterized by over-expression of the mdr1 gene product P-glycoprotein and cross-resistance to multiple classes of anticancer drugs, including quinones, seriously complicates and compromises the effective chemotherapy of cancer. For example, as compared to their parental counterparts, many MDR cell lines are over 10-fold more resistant to the well known antineoplastic agents doxorubicin (DOX) and etoposide (ETP). Accordingly, there is a constant search for antineoplastic agents that are not MDR substrates and which are therefore capable of effectively treating MDR cancers.
Naturally occurring substances are a promising source for such agents. Blackseed, the seed of
Nigella sativa
L. [Ranunculaceae], has been employed for thousands of years as a spice and food preservative, as well as a protective and curative remedy for numerous disorders. The historical tradition of blackseed in medicine is substantial. It is the blackseed referred to by the prophet Mohammed as having healing powers; blackseed is also identified as the curative black cumin in the Holy Bible, and is described as the
Melanthion
of Hippocrates and Doscorides and as the
Gith
of Pliny. Several beneficial pharmacological effects have been attributed to various crude and purified components of blackseed, including antihistaminergic, antihypertensive, hypoglycemic, antimicrobial, mast cell stabilizing and anti-inflammatory activities.
Blackseed preparations may have significant in vitro and in vivo antineoplastic activity. A crude methanolic extract of
Nigella sativa
seed has an in vitro IC
50
to Erlich ascites carcinoma, Dalton's ascites lymphoma and sarcoma 180 cells of 1.5, 3 and 1.5 &mgr;g/mL, respectively, while exerting minimal cytotoxicity to normal lymphocytes. This extract is also active in vivo, completely inhibiting the growth of Erlich ascites carcinoma in mice. Exposure to the volatile oil obtained from blackseed is believed to alter the cellular expression of specific polypeptides in Jurkart T lymphoma cells, suggesting that changes in polypeptide expression might play a role in the biological activities attributed to blackseed. In addition to these direct anti-tumor effects, blackseed preparations may have potential for cancer chemoprevention, as well as for reducing the toxicity of standard antineoplastic drugs. Topical application of a blackseed extract can inhibit the two-stage initiation-promotion (by dimethylbenz[a]anthracene-croton oil) of skin carcinogenesis in mice. Intraperitoneal injections of the same blackseed extract may reduce the incidence of soft tissue sarcomas observed after 30 days of subcutaneous 20-methylcholanthrene (MCA) injections by 67% as compared to MCA-treated controls. Another chemoprotective effect is treatment with a crude ethanolic extract of blackseed, which may significantly reduce cisplatin-induced leukopenia and hemoglobinemia in cisplatin-treated mice.
Although the above applications illustrate the antineoplastic potential of crude blackseed preparations, they do not establish the anti-cancer activity of specific blackseed components, in particular that of the quinones thymoquinone (TQ) and dithymoquinone (DIM). Further, no studies have been reported which evaluate the cytotoxicity of the
Nigella sativa
seed components TQ and DIM for multi-drug resistant cancer cells. Thus, comprehensive cytotoxicity assays of these potential anticancer drugs in both parental and MDR tumor cell lines are useful for a thorough evaluation and rational therapeutic development of these agents.
SUMMARY OF THE INVENTION
It is accordingly an aspect of the invention to provide methods and compositions for effectively treating both parental and multi-drug resistant cancers.
It is another aspect of the invention to provide methods and compositions, as above, which employ an extract from a natural source.
It is yet another aspect of the invention to provide methods and compositions, as above, which employ an extract from a natural source which is inexpensive and readily available.
These objects and others set forth below are achieved by treating cancer in an individual in need of such treatment by administering to said individual an antineoplastic amount of a compound selected from the group consisting of thymoquinone, dithimoquinone, and mixtures thereof.
To establish antineoplastic and cytotoxic activity, a crude gum, fixed oil and two purified components of
Nigella sativa
seed, TQ and DIM, were assayed in vitro for their cytotoxicity for several parental and multi-drug resistant (MDR) human tumor cell lines. Although as much as 1% w/v of the gum or oil was devoid of cytotoxicity, both TQ and DIM were cytotoxic for all of the tested cell lines IC
50
's 78 to 393 &mgr;M. Both the parental cell lines and their corresponding MDR variants, over 10-fold more resistant to the standard antineoplastic agents doxorubicin (DOX) and etoposide (ETP), as compared to their respective parental controls, were equally sensitive to TQ and DIM. The inclusion of the competitive MDR modulator quinine in the assay reversed MDR Dx-5 cell resistance to DOX and ETP by 6- to 16-fold, but had no effect on the cytotoxicity of TQ or DIM. Quinine also reduced MDR Dx-5 cell accumulation of the P-glycoprotein substrate
3
H-taxol in a dose-dependent manner. However, neither TQ nor DIM significantly altered cellular accumulation of
3
H-taxol. The inclusion of 0.5% v/v of the radical scavenger DMSO in the assay reduced the cytotoxicity of DOX by as much as 39%, but did not affect that of TQ or DIM. This suggests that TQ and DIM, which are cytotoxic for several types of human tumor cells, may not be MDR substrates, and that radical generation may not be critical to their cytotoxic activity.
REFERENCES:
patent: 4704384 (1987-11-01), Driscoll et al.
patent: 4922901 (1990-05-01), Brooks et al.
patent: 5482711 (1996-01-01), Medenica
patent: 5653981 (1997-08-01), Medenica
Johnson et al., Nat. Prod. Lett., 11(4), 241-250 Abstract Only, 1998.*
David R. Worthen et al., “The In Vitro Anti-tumor Activity of Some Crude and Purified Components of Blackseed,Nigella Sativa L., ” Anticancer Research, vol. 17, 1997, pp. 1-6.
N.J. Salomi et al., “Antitumour Principles fromNigella sativaseeds,”Cancer Letters, vol. 63, 1992, pp. 41-46.
M.J. Salomi et al., “Inhibitory Effects ofNigella sativaand saffron (Crocus sativus) on Chemical Carcinogenesis in Mice,” Nutr. Cancer, vol. 16, issue 1, pp. 67-72.
El-Mofty et al., “Prevention of Skin Tumors Induced by 7,12-dimethylbenz (a) anthracene in Mice by Black Seed Oil,”Oncology Reports, vol. 4, issue 1, 1997, pp. 139-141.
R. Medenica et al., “Immunomodulatory and Anti-Cancer Activity ofNigella sativaPlant Extract in Humans,” Proceedings of the American Association for Cancer Research, vol. 35, Mar. 1994, p. 481.
Crooks Peter A.
Ghosheh Omar A.
Worthen David R.
Goldberg Jerome D.
McDermott & Will & Emery
University of Kentucky Research Foundation
LandOfFree
Use of the naturally-occurring quinones thymoquinone and... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Use of the naturally-occurring quinones thymoquinone and..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Use of the naturally-occurring quinones thymoquinone and... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2486459