The Field of Invention

The present invention related to a process for preparing ethanol extract of plant ( Cuscuta campestris ) to the ethanol extract thus obtained, treatment of colon cancer.

The Background of the Present Invention

Colon cancer is one of the most common causes of death. Cancer is still one of the most important health problems. The purpose of the present invention is to produce an extract for colon cancer treatment. The improvements made about the subject in the related technical field are given below. Ethanol, water, and methanol extract of other Cuscuta species were often studied and exhibited potential anticancer properties on some cancer cell lines. For instance, the study of Suresh et all., (201 1) showed that the water extract of C. reflexa had dose dependently toxicity on the proliferation of Hep3B cells with no significant toxicity on the non- transformed cell line. Another study investigated the anti-proliferative potential of 95 % alcoholic, 50 % hydro alcoholic and water extracts along with n-hexane, chloroform, n-butanol and water fractions of 95% ethanolic extract of C. reflexa Roxb. The alcoholic extract and its chloroform fraction were found to be most active among three extracts and four fractions of alcoholic extract of whole plant of C. reflexa showed maximum cytotoxicity against human breast cancer cell lines (MCF-7). The alcoholic extract showed significant tumor growth inhibition for Ehrlich tumor and Sarcoma-180 solid tumor model (Bhagat, Arora, and Saxena 2013).

However, no study has been found about that Cuscuta campestris exhibits anticancer properties on colon cancer diseases.

Brief Description of The Present Invention

The present invention relates to Cuscuta campestris ethanol extract used in the treatment of colon cancer diseases in the health sector. The present invention provides a method for preparing ethanol extract of Cuscuta campestris treatment of colon cancer. The invention particularly relates to the use of the extract obtained from Cuscuta campestris in the treatment of colon cancer diseases.

The purpose of the present invention is to provide treatment of colon cancer diseases with the extract obtained from Cuscuta campestris.

A purpose of the present invention is to develop a medicament for use in the treatment of colon cancer, which is easy to produce and administer, and which does not have any adverse effect.

Another purpose of the invention is to provide an ethanol extract having anticancer activity, obtained with extraction method from Cuscuta campestris.

Another purpose of the invention is to obtain Cuscuta campestris ethanol extract, which is not found in mixture form and can be used alone in cancer treatment.

In order to achieve the above said purposes, the invention consists of Cuscuta campestris ethenol extract enabling treatment of cancer diseases.

In order to achieve the purposes of the invention, said extract is an extract obtained from Cuscuta campestris via extraction method in the presence of a solvent, solvent include ethanol solvent.

In order to achieve the above said purposes, the invention is the use of Cuscuta campestris extract in cancer treatment.

The figure and the detailed description given below in reference to the figure.

Detailed Description of Present Invention

As mentioned above, the first aspect of the invention relates to a method for preparing ethanol extract of Cuscuta campestris. The present invention relates to an extract prepared for use in the treatment of colon cancer diseases. Ethanol extract shows anticancer activity. Cuscuta campestris is holoparasitic plants from Convolvulaceae family .Cuscuta campestris Yuncker with the common name dodder has mainly leafless, smooth, yellow to pale orange twining stems about 0.3 mm in diameter. Cuscuta species, has been a research subject in a number of study for its bioactivities and bioactive compounds. Flavonoids, polysaccharides, alkaloids, steroids, volatile oils and lignans were found as the main compounds of Cuscuta species (Du et al. 1998; Miyahara et al. 1996; Shekarchi et al. 2014; Wang et al. 2000).

Extraction Procedure

The Preparation Method of the Cuscuta campestris Extract According to The Invention is Given Below

Cuscuta campestris plant was collected from Talas, Kayseri (Turkey), which is 1400 m in altitude, in August 2018. The plant samples were dried as a whole at room temperature and ground using a blender. The resulting powder was divided into balloon, 2 g/balloon. Sample was defatted with 10 mL of n-hexane at 30°C for 10 minutes. Ethanol extractions of residue were performed with 2 x 10 mL of ethanol at 60°C for 90 min in ultrasonic bath. Extraction was repeated twice at same extraction condition. The extracts was dried by rotary evaporator at 40°C.

Cell Cytotoxicity Test

The human colon carcinoma cell line (CACO-2) was used for cytotoxicity test. The in vitro cytotoxicity of ethanol extract to assess using ISO 10993-5: describes test methods. ISO (The international Organization for Standardization) is a worlwide federation of national standards. When analyzing data according to ISO standard, the sample has cytotoxic potential if the viability is below 70%. The in vitro study for the cytotoxic effect of the extract on CACO-2 human colon carcinoma cell line indicated that the ethanol extract has potential anticancer activity.

The human colon (colorectal adenocarcinoma) cell line (CACO-2) was grown in RPMI-1640 medium supplemented with 10 % FBS at 37°C in a humidified incubator with 5% CO2. Cell growth inhibition assessment in the presence of the extracts was performed essentially as described by ISO 10993-5. First day, cell suspension of CACO-2 (IxlO ⁴ cells mL ¹ ) was seeded 100 pL/well of a 96-well plate and incubated for 24 h at 37°C in a humidified incubator with 5% CO2. Next day, the growth medium was decanted and cells were incubated with serial dilutions of the stock solutions of extracts in dimethyl sulphoxide (DMSO) to adjust at 1000, 500, 250, 125, 62.5, 31.25, 15.63, 7.81, 3.91, 1.95, 0.98,. and 0.49 pg.mL ¹ concentrations in triplicates (100 pL/well) and incubated for 24 h at 37°C in a humidified incubator with 5% CO2. Cisplatin was used as positive control for cell cytotoxicity. After 24 h incubation of the treated and untreated cells (negative controls) at 37°C, 10 pL of 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium Bromide (MTT) solution 50 pL (1 mg.mL ¹ ) concentration was added in each well and incubated at 37°C for 2 h for the living cells to metabolize and convert soluble MTT salt into purple formazan crystals. The medium was then replaced with 100 pL DMSO per well. After 30 minutes, the absorbance of formazan crystals was determined by measuring the optical density (OD) at 570 nm using a Multiwall spectrophotometer (Tecan, Sunrise, Switzerland). The percentage of cell cytotoxicity for each concentration of the extracts was calculated by the following formula Viab. %=(Test OD/Control OD) x 100.

Description of Figures

Figure 1. Cell viability (%) of cytotoxic effect on CACO-2 colon carcinoma cell line treated with ethanol extract of Cuscuta campestris

Result

Consequently, the invention is obtained by using ethanol via extraction method. The extract of Cuscuta campestris obtained by dissolving with ethanol acts by reducing the number of CACO- 2 colon cancer cells in the cell culture. The above-mentioned study shows the anticancer activity of Cuscuta campestris. Said anticancer activity can be obtained from using the ethanol extracts of Cuscuta campestris. The extract is obtained by separating the substances found within Cuscuta campestris with the help of a solvent via extraction method.