DESCRIPTION (provided by applicant): Biologically active, non-steroidal, androgenic and estrogenic agents are found primarily in soy products, legumes, and whole grains, as well as in medicinal teas and roots. Kola acuminate, also known as Bizzy nut to the Ettu people of Jamaica, is a "cure-all" herbal medicine that reportedly affects a variety of biological processes,
many of which are directly or indirectly modulated by hormones. Available ethnobotanical information suggests that Kola acuminate may contain bioactive chemicals that possess estrogenic and androgenic properties. Anecdotal reports suggest that Bizzy nut may be useful for a number of medical purposes. Given the hormonal dependency of some of these biological activities, it is possible that non-steroidal androgen present in the Bizzy nut is responsible for he medicinal value attributed to it. The key to an agent's success, as a chemo preventive agent in the prostate and other types of cancer, relies on the agent's ability to induce tumor cell death in
a tissue-dependent and receptor-dependent manner. Naturally occurring agents that induce apoptosis in prostate cells in an AR-dependent manner would signal a new generation of prostate-specific, chemo preventive agents. The goal of this research project is to test the hypothesis that ether extract of Bizzy (Biz-2) modulates prostate function in a predictable manner. Thus, the overall objective of this revised application remains the same, namely demonstrating that the Kola acuminate (Bizzy Nut) extract contains bioactive compounds capable of functioning as chemo preventive agents against prostate cancer. The initial focus of this application is to isolated and identify the bioactive compounds found in the Biz-2Fr.3 fraction of Bizzy Nut and demonstrate, In vitro, its anti- cancer/chemo preventive potential using three prostate cancer cell models. To this end, two Specific Aims were developed. Aim 1: is to isolate and characterize the bioactive compounds in Bizzy nut extract associated with its anti-cancer properties in prostate cells. The goals of this Aim are to (1) Isolate and characterize the bioactive components of Biz-two by preparative HPLC; (2) elucidate the structure of the compounds in Biz-2Fr.3 using a combination of UV-Vis, NMR spectroscopy, LC-MS; and (3) Demonstrate the anti-cancer capability of the bioactive substances in Biz-2Fr.3. Aim 2: is to Identify the Biz-2Fr.3-induced apoptotic signaling pathways and determine the molecular mechanisms by which Biz-2FR.3 induces apoptosis in prostate cells. The goals of this Aim are to; (1) determine the mechanism of Biz-2Fr.3 anti-tumor activity using change in cell cycle distribution and protein expression profiling of cyclines. (2) Determine the involvement of mitochondria in Biz-2Fr.3 induction of apoptosis in normal and cancerous prostate cells using three prostate cell lines, the AR+ LNCaP, the AR- DU145 cells and RWP-1a normal, transformed cell line. We are well prepared to undertake the proposed research because of our experience with the experimental systems (see preliminary results) and the unique team of expert scientists in chemistry, biology and biochemistry as well as our collaborators and mentors from the LSU Chemistry and SUBR chemistry who are involved with the project. We have developed and assembled a set of tools, critical reagents and cell lines that will ensure success of this application. In addition, the proposed study is strongly supported by our preliminary data that validate the hypotheses that guide the proposed research. Finally, the combination of the assembled expertise, adequate facilities, strong collaborations and institutional support will allow us to conduct this study in a research environment that is conducive to its success.