the present invention may be described by reference to the figures . several varieties of cancer cells were grown in laboratory conditions and tested with gö6976 and the results recorded . the experiments and the results are described below . cell cultures used the following cancer cells , which were supplied from the american type culture collection , rockville , md . : 3y1 rat fibroblast cells overexpressing the epidermal growth factor receptor ( egfr ). these cells were maintained in dulbecco &# 39 ; s modified eagle medium ( dmem ) supplemented with 10 percent bovine calf serum .) small cell lung cancer cells were grown in rpmi 1640 medium with 2 mm l - glutamine adjusted to contain 1 . 5 g / l sodium bicarbonate , 4 . 5 g / l glucose , 10 mm hepes , 1 . 0 mm sodium pyruvate , and 5 % fetal bovine serum . non - small cell lung cancer cells were grown in rpmi 1640 medium supplemented with 5 % fetal bovine serum , 5 ug / ml insulin , 10 ug / ml transferrin , 30 nm sodium selenite , 10 nm hydrocortisone , 10 nm beta - estradiol , 10 mm hepes , and 2 mm l - glutamine . hl60 human acute promyelocytic leukemia cells were grown in rpmi 1640 medium containing 10 percent heat - inactivated fetal bovine serum ( fbs ). for growth of these cells in soft agar , 1 × 10 3 cells were suspended in top agar ( consisting of 20 percent calf serum , 0 . 38 percent agar and the remainder dmem ) and overlaid onto hardened bottom agar ( dmem , 20 percent calf serum and 0 . 7 percent agar ) as reported by qureshi et al ., in biochem . biophys . res . commun . 192 : 969 - 975 . cells were seeded for 24 hours . then they were either treated with gö6976 or left untreated . the cells were then collected periodically and evaluated using trypan blue dye exclusion to asses viability using the method described by cy chen , et al , in j . biol . chem . 273 : 16700 - 16709 . the results of treatment of human breast cancer cells ( mda - mb - 468 and mda - mb - 453 with gö6976 are shown in fig2 and 3 . cultures containing about 5 × 10 5 cells were treated with 0 . 0 , 0 . 5 , 1 . 0 and 2 . 0 micromole of gö6976 and the viable cells were counted at day 4 . as shown in fig2 cell count was reduced to about 1 × 10 5 when treated with 1 micromole of gö6976 and to less than 0 . 3 × 10 5 when treated with 2 . 0 micromolar of gö6976 . in a separate experiment as shown in fig3 the breast cancer cells were treated with 2 . 0 micromoles of gö6976 and measurements made at 24 hours intervals for 4 days . viable cell concentration appears to be reduced by approximately half each 24 hours . in both fig2 and fig3 error bars indicated the estimated standard error based on three independent experiments . the results of treatment of hl60 acute promyelocytic leukemia cells with gö6976 are shown in fig4 . one of two samples of 5 × 10 5 cells was treated with 2 micromoles of gö6976 and the viable cells were counted after four days . at four days the cells in the untreated sample had grown by a factor of nearly 10 to almost 5 × 10 6 cells whereas viable cells in the treated sample was less than 0 . 2 × 10 5 . the results of treatment of lung cancer cells are shown in fig5 . two samples each of about 5 × 10 5 cells small lung cancer cells and non - small lung cancer cells were prepared and one sample of each type was treated with gö6976 and the viable cells were counted at day 4 . in the untreated samples the cancer cells had multiplied by almost a factor of 10 and in the treated samples the count had decreased to less than 0 . 5 of the original count . the results of treatment of osteosarcoma cells are shown in fig6 . one of two samples of about 5 × 10 5 cells was treated with 2 micromoles of gö6976 and the viable cells were counted after four days . at four days the cells in the untreated sample had grown by a factor of nearly 8 to almost 4 × 10 6 cells whereas viable cells in the treated sample were reduced to about half the original count . the results of treatment of a431 human epidermoid carcinoma cells are shown in fig7 . one of two samples of about 5 × 10 5 cells was treated with 2 micromoles of gö6976 and the viable cells were counted each 24 hours for four days . at four days the cells in the untreated sample had grown by a factor of about 24 to almost 1 . 2 × 10 7 cells whereas viable cells in the treated sample had approximately doubled . fig8 shows the effect of gö6976 for blocking cell transformation caused by epidermal growth factor ( egf ). 3y1 cells overexpressing egf receptors showed a transformed morphology upon 100 nanamoles egf treatment for 24 hours . co - treatment with 250 nanamoles gö6976 and egf completely blocked egf - induced cell transformation while treatment with gö6976 alone did not show any toxic effect on 3y1 egfr cells . the effect of gö6976 on blocking egf - induced cell transformation was further confirmed by checking anchorage - independent cell growth . as shown in fig8 cultures of 1 × 10 3 3y1 cells overexpressing egf receptors were prepared by suspending the cells in soft agar . one sample was left untreated , one was treated with 100 nanomoles of egf , one sample was treated with 100 nanamoles of egf and 250 nanamoles of gö6976 and one sample was treated with 250 nanaomoles of gö6976 . in each case the percentage of cell forming colonies were determined three weeks after suspending the cells in soft agar . colony forming efficiency of egf was greatly reduced by gö6976 . applicant tested gö6976 on normal cells including rat 3y1 fibroblast in the concentrations used in the above - described experiments and no significant toxic effects on the cells were noted . toxic effects begin to show up at concentrations of about 5 micromoles in these types of cells . the above described test show clearly that gö6976 has a strong apototic effect on a variety of human cancer cells and that it can inhibit growth of other human cancer cells . based on the egf experiments showing a blocking of the effects of the egf induced cell transformation , gö6976 also can be used as a cancer preventive reagent . the lack of toxic effects on normal cells at the concentrations needed for effectiveness against the cancer cells shows that gö6976 is very valuable chemotheraputic agent . it could be applied in many of the well - known methods currently used for chemotheraputic treatment . for example , it may be injected directly into tumors , injected into the blood stream or taken orally or applied through the skin with patches . the dose chosen should be sufficient to constitute effective treatment but not so high as to cause unacceptable side effects . the state of the cancer and the health of the patient should preferably be closely monitored during and for a reasonable period after treatment . while the above examples describe the effectiveness of gö6976 as a chemotheraputic treatment for various cancers they are not intended as a limitation of the invention . the chemical was effective in killing or reducing the growth rate of all cancer cells tested . therefore , it should be obvious that it would be effective as a treatment to cancer cells not tested that are associated with increased levels of pkc α expression and / or activity such as prostate cancer , pancreatic cancer and melanoma . derivatives of gö6976 that are effective in inhibiting pkc α should also be effective as a cancer treatment . also , since gö6976 prevents cell transformation it should also work as a preventative drug , especially for people having a high risk for particular cancers .
a chemotheraputic cancer treatment in which gö6976 or one of its derivatives is administered to a mammal for the treatment of the cancer . the gö6976 or its derivative is directed to pkc α activity . experiments have shown gö6976 to be effective for the treatment of breast cancer , leukemia , lung cancer , bone cancer and skin cancer . these treatments may be accomplished utilizing gö6976 or its derivatives alone or in combination with other prior art chemotherapy agents or in combination with radiation therapy . in a preferred embodiment gö6976 is used for the treatment of cancer as a preventative drug by preventing cancer cell formation .