GULER AKGEMCI EMINE (TR)
GULER AKGEMCI EMINE (TR)
| CLAIMS 1. An agent comprising Cu (P) complexes of 2-hydroxy-5-methoxyacetophenone thiosemicarbazone and N (4) -ethyl and N (4) -phenyl derivatives characterized in that having at least one of [Cu (HMAT) Cl], [Cu (HMAET) Cl] and [Cu (HMAPT) Cl] complexes. |
P HE NY L
TE C HNICAL FIE L D
The invention relates to an agent containing C u(II) complexes of 2- hydroxy- 5 metoxyacetophenone thiosemicarbazone and 4(N)-ethyl and n(4)-phenyl derivatives for supressing metastasis causing cancer death.
P RIOR ART
According to the World Health Organization (WHO), cancer incidance rates are predicted to increase by 70% over the next two decades. As a precaution, the existing methods need to be developed or changed to reduce these rates. If there is one thing worse than the formation of cancer that is the dispers of these cancer cells to other tissues. In the literature, the development of secondary malignant growths at a distance from a primary site of cancer described as ' metastasis _, and stopping this invasion has a vital importance. While the present medical treatment protocoles and chemotherapeutics do not give the desired response and also developed chemoresistance in some type of cancers, consequently new treatment approaches and protocols are needed in this field. The drugs which are used for treatment of the infections caused by bacteria, protozoa, fungi, worms (helminths) or viral induced neoplastic transformations could be used as adjuvant chemotherapeutic agents. Nevertheless, according to the current results the development of new drug molecules is essential in cancer.
B RIE F DE S C RIPTION OF THE INVE NTION
The invention is a metastatic inhibitor in the treatment of cancer with thiosemicarbazones, which can be found in antituberculous, antibacterial, antifungal, anticancer and antimalarial activities. The invention, is a novel compound formed with thiosemicarbazones for suppression of metastasis that causes cancer death. The component contains C u (II) complexes of the 2-hydroxy-5 methoxyacetophenone thiosemicarbazone ligand (HMAT), N (4) -Ethyl (H MAET), N (4) -Phenyl (HMAPT) derivatives. DETAILED DESCRIPTION OF THE INVENTION
Methods for producing Cu (II) complexes of the 2-hydroxy-5 methoxyacetophenone thiosemicarbazone ligand (HMAT), N (4) -Ethyl (HMAET), N (4) -Phenyl (HMAPT) derivatives are given below. i 2-HYDROXY-5-METHOXYASETOFE NONE THIOSOCE MARBAZONE (HMAT) PRODUCTION METHOD
- Mixing 1 mL (0.17grams) of 2-hydroxy-5-methoxyacetophenone with 30 mL of absolute ethanol and 1 mL (0.09 grams) of thiosemicarbazide in 20 mL of absolute ethanol at 100 mL.
- Adding 2-3 drops of concentrated sulfuric acid solution onto it,
- The mixture was stirred at room temperature for 2 hours,
- Next day the product formed by filtration,
- C rystallization occurs in the ethyl alcohol-water mixture in twice. i 2-HYDROXY-5-METHOXYAS ETOPHE NONE-N (4) -ETHYL
THIOCEMICARBAZONE (HMAET) PRODUCTION METHOD
- Mixing 1 mmol (0.17 g) of 2-hydroxy-5-methoxyacetophenone with 30 mL of absolute ethanol and 1 mmol (0.12 g) of 4-ethyl-3-thiosemicarbazide in 20 mL of absolute ethanol.
- Adding 2-3 drops of concentrated sulfuric acid solution onto it,
- The mixture was stirred at room temperature for 2 hours,
- Next day the product formed by filtration,
- Crystallization occurs in the ethyl alcohol-water mixture in twice. i 2-HYDROXY-5-METHOXY AS ETOPHE NONE-N (4) -PHENYL
THIOCEMICARBAZONE (HMAPT) PRODUCTION METHOD
- Mixing 1 mmol (0.17 grams) of 4-phenyl-3-thiosemicarbazide was dissolved in 30 mL of absolute ethanol and 1 mmol (0.17 grams) of 2-hydroxy-5- methoxyacetophenone was dissolved in 20 mL of absolute ethanol.
- Adding 2-3 drops of concentrated sulfuric acid solution onto it
- The mixture was stirred at room temperature for 2 hours,
- Next day the product formed by filtration, Crystallization occurs in the ethyl alcohol-water mixture in twice. i 2-HYDROXY-5-METHOXYACETOPHE NONE THIOCE MICARBAZON COPPER (II) COMPLEX PRODUCTION METHOD
- Dissolving 1 mmol (0.24 grams) of 2-hydroxy-5-methoxyacetophenone thiosemicarbazone in 30 mL of ethanol by heating,
- Dissolving 1 mmol (0.17 g) of copper (II) chloride dihydrate (CuCI2.2H20) in the minimum amount of the same solvent,
- Addition of this solution dropwise to the 2-hydroxy-5-methoxyacetophenone thiosemicarbazone solution,
- The mixture is refluxed for about 2 hours,
- The appropriate solid phase is slowly evaporated at35eC,
- Filtration of the obtained layer,
- Washed with anhydrous ether,
- Drying at room temperature. i 2-HYDROXY-5-METHOXYASETOFE NON N (4) -ETHYL THIOCE MICARBAZON COPPER (II) COMPLEX PRODUCTION METHOD
- Dissolving 1 mmol (0.27 grams) of 2-hydroxy-5-methoxyacetophenone N (4) - ethyl thiosemicarbazone in 30 mL of ethanol,
- Dissolving 1 mmol (0.17 g) of copper (II) chloride dihydrate (CuCI2.2H20) in the minimum amount of the same solvent,
- Addition of this solution dropwise to the solution of 2-hydroxy-5- methoxyacetophenone N (4) -ethyl thiosemicarbazone,
- The mixture is refluxed for about 2 hours,
- The appropriate solid phase is slowly evaporated at35eC,
- Filtration of the obtained layer,
- Washed with anhydrous ether,
- Drying at room temperature. i 2-HYDROXY-5-METHOXYACETOPHE NONE N (4) -PHENYL THIOCEMICARBAZON COPPER (II) COMPLEX PRODUCTION METHOD
- Dissolving 1 mmol (0.32 grams) of 2-hydroxy-5-methoxyacetophenone N (4) - phenyl thiosemicarbazone in 30 mL of ethanol,
- Dissolving 1 mmol (0.17 g) of copper (P) chloride dihydrate (C uC I2.2H20) in the minimum amount of the same solvent,
- Addition of this solution dropwise to the 2-hydroxy-5-methoxyacetophenone thiosemicarbazone solution,
- The mixture is refluxed for about 2 hours,
- The appropriate solid phase is slowly evaporated at 35eC,
- F iltration of the obtained layer,
- Washed with anhydrous ether,
- Drying at room temperature.
The aim of this study is to identificate the effectiveness of our invention, to exemplificate without any restrictions, and to determinate the cytotoxic effect of the Cu (II) complexes of 2-hydroxy-5 methoxyacetophenone thiosemicarbazone ligand (HMAT) and N (4) -ethyl (HMAET) and N (4) -phenyl (HMAPT) derivatives on MDA- MB-231 cells, a highly metastatic and chemo-resistant breast cancer cell line. The cytotoxicity assay results showed that, the in vitro IC so values of our complexes ranged from 1.76-3.53 =M, and this means the C u(II) complexes showed at least 20 times higher activity than its ligands.
The anti-metastatic effect of these new complexes we synthesized ([C u (HMAT) C l], [C u (HMAET) C l] and [C u (HMAPT) C l]) was also determined by Western blot. In our study, we evaluated the epithelial mesenchymal transition progression levels (by its markers; E -Cadherin, N-Cadherin and Vimentin) of highly invasive MDA-MB-231 cells in conditions at defined IC50 values of the complexes. According to our results, three of the our complexes increased the expression of E -Cadherin in MDA-MB 231 cells. The fact that the [C u (HMAPT) C l] complex increased E -cadherin expression by 300% atthe highest rate. This helps to suppress the metastatic ability of the cells. Also the three of our complexes suppressed N-Cadherin by 50-70%. This result shows that the invasion ability of the cell is reduced. In addition, all of our complexes suppress expression of Vimentin which is cytoskeletal component supporting the cell invasion. E specially [C u (HMAET) C l] reduced Vimentin expression by 90% and stopped the ability of the cell to metastasize. In addition to this, we investigated changing the expression level of TWIST1 which is the key molecule of epithelial mesenchymal transition process and endogenously highly expressed in MDA_MB-231 cells, in the presence of our complexes in vitro. According to the results, all of our new complexes suppressed the expression of TWIST1. It was determined that [Cu (HMAET) Cl] complex was suppressed TWIST1 by 80% at most. Finally, these findings showed that the newly synthesized [Cu (HMAT) Cl], [Cu (HMAET) Cl] and [Cu (HMAPT) Cl] complexes could be evaluated as novel anti-cancer and anti-metastatic agents.