Indonesia nanopropolis as a antibreastcancer agents

Abstract

Propolis is a honeybee product that is useful for self-protection, especially from invading microorganisms and temperature changes outside the beehive. Today, the term drug dosage propolis is extracted from beehives called raw honey or propolis. Raw propolis can not be used directly, so it is necessary that the process of extracting propolis is a mixture of active ingredients using organic solvents. The active ingredients of propolis were flavonoids, alkaloids, tannins, steroids and triterpenoids. Raw propolis is extracted by 70% ethanol to obtain propolis flavonoid-containing components of raw propolis as a whole and separate from the balm. There have been many studies using maceration extraction, but the combination of maceration and heating by microwaves has not been done. Therefore, the process of separation of propolis from the beehive (raw propolis) by using maceration and microwave heating was undertaken in this study. Many studies have shown that β-cyclodextrin can be used as a coating agent active ingredient in the drug manufacturing process. Circular shape consisted of seven glucose monomers, β-cyclodextrin can be inserted and bind the active ingredient in various side active. With the inclusion of the active ingredient in β- cyclodextrin group then expected release of the active ingredient will be much slower than the free active ingredient. Therefore, the inclusion of propolis research on β-cyclodextrin, so that the active ingredient in propolis out of filler slowly. To make the coating process of propolis more effectiveing the process was carried out by a high speed homogenizer. Therefore, this research was undertaken propolis inclusion in β-cyclodextrin using inclusion, resolubility and stabilisation with high speed homogenization as processing aid. With the inclusion, resolubility and stabilisation process was expected that there were active ingredient of propolis is adsorbed molecule β-cyclodextrin and particle size reduction occurs. In addition, the amount of propolis and β-cyclodextrin is used in the process of shaking on the inclusion process may be a factor in the rate of inclusion process, so there are differences in the amount of active ingredient present in propolis adsorbed in β-cyclodextrin. Adsorbed amount of active ingredient and the ability to withstand damage due to the active ingredient high speed could be demonstrated by the inhibition of cancer cell proliferation activity of Michigan Cancer Foundation-7 (MCF-7). Nanopropolis preparation process was performed by inclusion, resolubility and stabilitation with high speed homogenization (22000 rpm) at the process of making nanoparticles. The first stage times were 20, 30 and 40 mins, the second times were 20, 30 and 40 mins, and the third times were 10, 20 and 30 mins. Nanopropolis were researched their particle distribution and effectiveness as an anticancer ingredient. The data obtained were analyzed by Response Surface Methodology (RSM) to determine the best conditions for making nanopropolis. At the best conditions (of the inclusion process) then undertaken nanopropolis preparation with propolis and variable number of β-cyclodextrin. The results were tested on the inhibition of proliferation of MCF-7 cancer cells and performed the statistical analysis with the RSM to determine the best conditions for making nanopropolis. Further efficacy testing was carried out using test animals nanopropolis female white rats of Sprague Dawley strain induced by 7,12- dimethylbenz(a)anthracene (DMBA). After in-vivo study ended, the development of tumors in rats was studied by looking preparated mammary tissue under microscope. Propolis from five locations in Indonesia showed the difference in extraction yield (% w/w), total flavonoid content (%), the ability to inhibit the oxidation of DPPH (IC50, μg ml-1), apoptosis induction for Saccharomyces cerevisiae cells at concentrations of 50 mg ml-1, and inhibits cell proliferation sustainable MCF-7 breast cancer at a concentration of 100 μg ml-1. The best results from yield of propolis extract was obtain from Pekanbaru, total flavonoid content was from Kendal, free radical scavenging of 1,1-diphenil -2-picrilhydrazil (DPPH) was from Pandeglang, Saccharomyces cerevisiae induces cell apoptosis was from Kendal, and inhibits proliferation of breast cancer cells MCF-7 was from Makassar with the value of 19.97 (%), 46.60 (%), 68.94 (μg ml-1), 81.44 (%), and 47.71 (% living cells), respectively . All propolis extracted from five locations in Indonesia based on phytochemical analysis showed that the propolis contains flavonoid compounds. Results of statistical analysis showed that induction of apoptosis against S. cerevisiae cells as much as 85% was achieved at the best conditions of microwave heating time of 30 mins and the ratio of 70% ethanol-beehive of 20. Results of the verification process of extraction conditions selected showed that the percentage of cells petite was 70.32% smaller than the model, but the yield was higher than the models. Propolis Trigona spp originated Pandeglang had antioxidant activity (IC50) of 75.34 μg ml-1, 50% lethal cancer MCF-7 cells at a concentration of 233 μg ml-1, with a value of cell apoptosis induction S. cerevisiae of 6.02 μg ml-1. The RSM analysis results nanopropolis predicted that the best conditions was propolis of 30 mg to 350 mg of β-cyclodextrin with IC50 at 10.2 μg ml-1. At the best conditions, the propolis could cause the death of MCF-7 cells by as much as 48.61% and a yield of 18.657 g. Nanopropolis generated by inclusion in β- cyclodextrin showed that the change in crystallinity from 31% to 88.7% still approved flavonoids and organic acids on High Performance Liquid Chromatography (HPLC) analysis especially techtochrysin and caffeic acid; on Fourier Transform Infra Red (FTIR) analysis there has been a change in wave number, mainly on the functional group-OH. Nanopropolis average particle size distribution produced was 171 nm. . In the in-vivo test, propolis demonstrated that it could heal damaged tissue tumors at a concentration of 233 μg ml-1. At concentrations of 32 and 56 μg ml-1 nanopropolis already showed the results of healing damaged tissue tumors.