DESC:FIELD OF THE INVENTION
The present invention relates to anticancer composition and in particularly relates to the anticancer composition with lesser side effects.

BACKGROUND OF THE INVENTION
Chemotherapy is one of the most common treatments for cancer. It uses certain drugs to kill cancer cells or to stop them from growing and spreading to other parts of the body. Even after surgery to remove a tumor, body might still have cancer cells. These cells can grow new tumors or spread the cancer to other parts of your body. Chemotherapy drugs help destroy, shrink, or control those cells. Chemotherapy drugs can kill both cancerous and healthy cells, fight only cancer cells, keep tumors from growing blood vessels which help them thrive or attack the cancer cell’s genes so the cells die and can’t grow into new tumors.

Chemotherapy resistance occurs when cancers that have been responding to a therapy suddenly begin to grow. In other words, the cancer cells are resisting the effects of the chemotherapy. When this occurs, the drugs will need to be changed. Research is underway to investigate ways of reducing or preventing chemotherapy resistance. The development of drug resistance is one major reason that drugs are often given in combination. Often, if a cancer becomes resistant to one drug or group of drugs, it is more likely that the cancer may be resistant to other drugs. This is why it is very important to select the best possible treatment protocol FIRST. Chemotherapy fails to cure most cancer patients with advanced disease, particularly patients with the most common forms of solid tumors. The presence or development of resistance to anticancer agents is the major cause of this failure.

Thus, there emerges a great need for an anticancer composition that has less side effects and reduced development of drug resistance.

OBJECTIVES OF THE INVENTION
It is an object of the present invention to provide an anticancer composition, useful for chemotherapy.

Another object of the invention is to provide an anticancer composition acts with reduced side effects of chemotherapeutic drug Cisplatin.

Yet another object of the invention is to provide an anticancer composition that minimizes the development of drug resistance.

Yet another object of the present invention is to provide an anticancer composition that utilizes lesser dose of Cisplatin with more efficacy.

SUMMARY OF THE INVENTION
The present invention provides an anticancer composition, comprising Cisplatin and Planispine A. The anticancer composition is useful for killing cancer cells during chemotherapy and utilizes lower concentration of Cisplatin with greater efficacy.

Further, the present invention discloses an anticancer composition that has reduced side effects and minimized development of drug resistance. The present anticancer composition comprising Planispine A and Cisplatin augments its apoptotic potential and postulates a chemo-preventive approach in ovarian cancer cells.

BRIEF DESCRIPTION OF THE DRAWINGS
For a better understanding of the embodiments of the methods described herein, and to show more clearly how they may be carried into effect, references will now be made, by way of example, to the accompanying drawings, wherein like reference numerals represent like elements/components throughout and wherein:

Figure 1 illustrates different steps of the process of extracting Planispine A, according to an embodiment of the present invention.

Figure 2 shows the FTIR spectrum of Planispine A, according to an embodiment of the present invention.
Figure 3 shows the 1H NMR of Planispine A, according to an embodiment of the present invention.

Figure 4 shows the 13C NMR of Planispine A, according to an embodiment of the present invention.

Figure 5 shows the synergistic interaction with chemotherapeutic drug Cisplatin, according to an embodiment of the present invention.

Figure 6 shows the flow cytometric analysis of combination of Planispine A and Cisplatin on cell cycle, according to an embodiment of the present invention.

DETAILED DESCRIPTION OF THE INVENTION
The embodiments of the invention will now be described herein, with reference to the accompanying examples and drawings. It will be understood by those skilled in the art that the foregoing general description and the following detailed description are exemplary and explanatory of the invention and are not intended to be restrictive thereof.
The present invention provides anticancer composition comprising Cisplatin and Planispine A.
According to an embodiment of the present invention, the anticancer composition includes Planispine A and Cisplatin in the ratio 1:5. The said anticancer composition provides a synergistic anticancer effect of Planispine A that enhances the therapeutic potential of chemotherapeutic drug Cisplatin.
In an embodiment of the invention, the said Planispine A has been extracted from the leaves of Zanthoxylum armatum DC (Rutaceae). The extraction process 100 as illustrated in Figure 1 begins at step 101, wherein the leaves of Zanthoxylum armatum DC (Rutaceae) were collected, washed and dried in shade at room temperature. At step 102, dried leaves were grounded into fine course. Further at step 103, grounded leaves were soaked in methanol for 2-3 days. The extract was filtered through Whatman filter paper No. 1 at step 104 and concentrated using rotatory vacuum evaporator (Methanol vac. Pressure 337 mbar at 40°C) under reduced pressure. The step 104 is repeated thrice for maximum yield. At step 105, extract is subjected to column chromatography with ethyl acetate followed by 90 – 10% hexane to 10 – 90 % ethyl acetate gradients. At step 106, bioactive fractions are further purified by reverse phase C-18 column chromatography, HPLC to obtain pure compound. Further at step 107, molecular structure of the compound (Planispine A) is identified by NMR (1D and 2D), mass spectroscopy and FTIR analysis.
In another embodiment of the present invention, the anticancer composition has apparent inhibitory effect on cell proliferation as compared to the inhibitory effect observed by application of the individual drugs Cisplatin and Planispine A. Thus, the synergistic effect shows anticancer activity with more efficacy at lower concentration of Cisplatin that might provide a new therapeutic strategy, reducing its side effects and development of drug resistance.
In yet another embodiment of the present invention, the concentration of Planispine A is 9-11 µM and of Cisplatin is 49-51 µM.
In yet another embodiment of the present invention, the anticancer composition is useful for killing cancer cells during chemotherapy and utilizes lower concentration of Cisplatin with greater efficacy.
In yet another embodiment of the present invention, the anticancer composition augments its apoptotic potential and postulates a chemo-preventive approach in ovarian cancer cells.
EXAMPLES
In an embodiment of the invention, the composition comprising Planispine A and Cisplatin was checked for its effectiveness to synergistically induce apoptosis. PA1 cells were pre-treated with the Planispine A (2 and 10 µM) for 16 h followed by treatment with Cisplatin (25 and 50 µM) for 72 h. Viable cells were measured and Q value more than 1.15 according to Jin’s formula were used for the representation (indicated with asterisk in graph). It was observed that Planispine A shows synergistic interaction with chemotherapeutic drug Cisplatin (Figure 5).

In another embodiment of the invention, the composition comprising Planispine A and Cisplatin was checked for its effectiveness to arrest S phase of the cell cycle. Cells were treated with the indicated Planispine A or Cisplatin alone or combination. Cells were fixed with 70% ethanol, stained with Propidium iodide and subjected to flow cytometry for determining the cell cycle. It was observed that anticancer composition leads to massive S phase arrest (Figure 6).
The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the disclosure. It will be apparent to those skilled in the art that various modifications and variations can be made in the products and methods of the present invention without departing from the spirit or scope of the invention. Thus, various presently unforeseen or unanticipated alternatives, modifications, variations, or improvements therein may subsequently be made by those skilled in the art without departing from the scope of the present disclosure as encompassed by the following claims. Additionally, the foregoing examples are appended for the purpose of illustrating the claimed invention, and should not be construed so as to limit the scope of the claimed invention.
,CLAIMS:We claim:
1. A synergistic anticancer composition that minimizes the development of drug resistance comprising:
a) Cisplatin; and
b) Planispine A.

2. The synergistic anticancer composition as claimed in claim 1, wherein the amount of cisplatin and planispine A is 5:1.

3. The synergistic anticancer composition as claimed in claim 1, wherein the concentration of Planispine A is 9-11 µM and of Cisplatin is 49-51 µM.

4. The synergistic anticancer composition as claimed in claim 1, wherein the Planispine A is obtained from the leaves of Zanthoxylum armatum DC.

5. The synergistic anticancer composition as claimed in claim 1, wherein the said anticancer composition imparts inhibitory effect on cell proliferation.

6. The synergistic anticancer composition as claimed in claim 1, wherein the process for preparing the Planispine A (100) comprising the steps of:
a) Collecting, washing and drying the leaves of Zanthoxylum armatum;
b) grinding the leaves;
c) soaking ground leaves in methanol for 2-3 days;
d) filtering and concentrating the extract;
e) performing column chromatography with ethyl acetate followed by 90 – 10% hexane to 10 – 90 % ethyl acetate gradients;
f) purifying bioactive fractions by reverse phase C-18 column chromatography and HPLC; and
g) identifying Planispine A by NMR, mass spectroscopy and FITR analysis.

7. The synergistic anticancer composition as claimed in claim 6, wherein the step d) is repeated thrice to obtain maximum yield.