PL228170B1 - Application of (E)-2-hydroxy-2',4'-dinitrostilbene derivatives in a pharmaceutical formulation - Google Patents

Description

The subject of the invention is the use of (E) -2-hydroxy-2 ', 4'-dinitrostilbene derivatives of formula 1,

Formula I wherein R-ι, R2, R3, R4 is hydrogen, methoxy or nitro, or a pharmaceutically acceptable form thereof, for the preparation of a pharmaceutical preparation for anti-neoplastic therapy, particularly glioblastoma and cervical carcinoma.

Stilbene derivatives are a group of chemical compounds containing in their structure two aromatic rings connected by two olefinic carbon atoms. The substituents on the phenyl rings determine the chemical, physicochemical and biological properties. The naturally occurring stilbenes are dominated by hydroxyl, methoxy and carboxyl substituents. High hopes are attached to the synthetically obtained stilbenes with strongly electron-withdrawing substituents.

Stilbene derivatives have extremely interesting biological properties. Naturally occurring resveratrol ((E) -3,5,4'-trihydroxystilbene) or pterostilbene ((E) -3,5-dimethoxy-4'-hydroxystilbene) are widely used in medicine and dietetics. These compounds show antioxidant, anti-carcinogenic and antifungal activity and slow down the aging process of the body. They prevent the development of coronary artery diseases and support the treatment of insulin resistance.

They also have estrogenic properties, which are why they are used in the prevention and treatment of complications in pregnancy.

The inhibitory effect of resveratrol on the development of neoplastic disease in its subsequent stages was also noted [B. Aggarwal, A. Bhardwaj, R.S. Aggarwal, N.P. Seeram, S. Shishodia and Y. Takada, Role of resveratrol in prevention and therapy of cancer: Preclinical and clinical studies. Anticancer Research, 24 5A (2004), pp. 2783-2840]. The use of resveratrol was found to reduce the incidence of skin cancer in mice by as much as 98%. Resveratrol has been shown to slow cancer tumor growth through multiple complementary mechanisms. It inhibits the enzymatic activity of both forms of cyclooxygenase, which reduces the risk of developing many cancers. Another mechanism by which resveratrol may help to slow disease progression is through disruption of the cell cycle, which often leads to apoptosis of cancer cells. The antiproliferative and pro-apoptotic effects of resveratrol in cell lines have been proven in vitro. However, in some research trials, no effect on the development of neoplastic disease was found, which may suggest an influence of the dosage, method of administration and type of neoplasm on the effectiveness of treatment.

It should be emphasized, however, that the action of stilbene derivatives is not always useful, sometimes even harmful. Such an example is diethylstilbestrol ((E) -4.4 '- (hex-3-en-3,4-olyl) diphenol). Based on the results of studies published in 1971, the use of preparations containing diethylstilbestrol in pregnant women was associated with the occurrence of various types of cancer in their offspring. These included cervical cancer and a vaginal tumor called adenoma [Herbst AL, Ulfelder H, Poskanzer DC. Adenocarcinoma of the vagina. Association of maternal stilbestrol therapy with tumor appearance in young women. The New England Journal of Medicine 1971; 284 (15): 878-881]. The biological activity of these compounds is closely related to the substituents that are attached to the aromatic rings or the double bond.

PL228 170 Β1

Therefore, new derivatives with a broader spectrum of activity against cancer are constantly sought.

Formula 1 wherein R ι, _R2, R3, R4 is hydrogen, methoxy or nitro, use 2-hydroxy-3-methoxy-2 ', 4'-dinitrostilben (JJR1), 2-hydroxy-4-methoxy 2' , 4'-dinitrostilbene (JJR5), 2-hydroxy-3nitro-2 ', 4'-dinitrostilbene (JJR6), 2-hydroxy-3,5-dimethoxy-2', 4'-dinitrostilbene (JJR9) cells were used against tumors human glioblastoma line U87 and human cervical carcinoma line HeLa cells.

The invention is described in more detail in the following embodiment.

Description and evaluation scheme of the antiproliferative activity of (E) -2-hydroxy-2 ', 4'-dinitrostilbene derivatives:

I. 2-hydroxy-3-methoxy-2 ', 4'-dinitrostilbene (JJR1)

II. 2-hydroxy-4-methoxy 2 ', 4'-dinitrostilbene (JJR5)

III. 2-hydroxy-3-nitro-2 ', 4'-dinitrostilbene (JJR6)

IV. 2-hydroxy-3,5-dimethoxy-2 ', 4'-dinitrostilbene (JJR9)

U87 human glioma cells and HeLa cervical carcinoma cells were cultured in vitro in 96-well plates at 3x10 ⁴ (HeLa) and 5x10 ⁴ (U87) plates per well in a volume of 100 µl DMEM culture medium supplemented with fetal bovine serum and antibiotics. After 24 hours incubation at 37 ° C in a humidified atmosphere saturated with CO _2, cells treated with these compounds at concentrations of 100, 50, 25, 12.5, 6.25 and 3.125 μΜ in a final volume of 150 μΙ. Stock solutions of test compounds at a concentration of 50 mM were prepared in dimethylsulfoxide (DMSO), dilutions were made in culture medium. The control cells were cells to which only the culture medium was added. After another 24 and 48 hours of incubation at 37 ° C, the degree of inhibition of proliferation of U87 and HeLa cells was determined. For this purpose, a fluorescence test with Alamar Blue reagent (Invitrogen) was used according to the manufacturer's instructions. On the basis of the plot of inhibition of cell proliferation on the concentration of the tested compound, IC50 values were determined - concentrations inhibiting the proliferation of HeLa and U87 cells in 50% (Table 1).

Table 1

IC50 values (pg / ml) obtained for the (E) -2-hydroxy-2 ', 4'-dinitrostilbene derivatives JJR1, JJR5, JJR6 and JJR9 against U87 and HeLa cells for different incubation times.

IC50 (pg / ml) HeLa IC50 (pg / ml) U87 24 h of incubation 48 h of incubation 24 h of incubation 48 h of incubation JJR1 53.32 ± 1.74 23.15 ± 3.59 - - JJR5 12.74 ± 1.99 5.05 ± 1.05 25.02 ± 2.44 17.18 ± 3.94 JJR6 10.82 ± 4.13 2.34 ± 0.48 49.64 ± 14.14 36.01 ± 5.22 JJR9 8.31 ± 4.03 6.21 ± 1.91 - 41.18 ± 21.77

Claims (1)

wherein R ι, _R2, R3, R4 is hydrogen, methoxy or nitro, or a pharmaceutically acceptable form, in the manufacture of a pharmaceutical preparation for anti-cancer therapy, in particular glioblastoma, and carcinoma of the cervix. Publishing Department of the PPO