5hrs with horseradish peroxidase conjugated secondary antibody. The protein bands were detected by a sophisticated chemiluminesence kit. Nude order ARN-509 mice xenografts Androgen dependent LNCaP and separate LNCaP AI prostate cancer cells, blended with Matrigel in a rate of 1:1 were inoculated to the flanks of 4 5 week-old male Nu/Nu Balb/c athymic nude mice by subcutaneous injection. Volume and the cyst growth was checked every 3 days. Animals were randomly split into 2 teams, 10 mice each, in accordance with tumor size, when the prostate tumor grew to a diameter of 4 8 mm. One group of animals was treated with drug car just as control, and still another group was treated with Natura leader at dose of 100mg/kg by gavage, once a day, 5 days per week before size of tumors in control group reached about 15 mm. The tumor growth was checked everyday and tumor size recorded every three days. The tumefaction size was determined as l x d x h x 0. 52. Proteomic Pathway Array Analysis Total cellular proteins were produced from tumors utilizing a lysis buffer containing 20mmol/L Tris HCl, 20mmol/L sodium Pyrophosphate, 40mmol/L T Metastatic carcinoma glycerophosphate, 30mmol/L Sodium Fluoride, 2mmol/L EGTA, 10mmol/L NaCl, and 0. Five full minutes NP 40. The lysate was sonicated three times for 15 seconds each time, and then centrifuged. The tubes were kept on ice through the process. The protein concentration was determined with the BCA Protein Assay kit. Remote proteins were separated by SDS PAGE. Three-hundred ug of protein extracts were loaded in a well throughout the whole thickness of gel for SDS PAGE, followed closely by electro transferring to some nitrocellulose membrane. The membrane was then blocked for 1 hr with five minutes milk or three or four BSA, and clamped on to a Mini PROTEAN II Multi-screen apparatus that isolates 20 routes across the membrane. 2 or 3 antibodies were included with each route and incubated over night at 4oC. Different sets of antibodies were employed for each membrane after stripping hdac1 inhibitor the last set of antibodies. Antibodies were acquired either from Cell Signaling Technology, Inc. or from Santa Cruz Biotechnology, Inc.. The pathway array analysis was run in duplicate for each sample in each pair of antibodies and protein levels were normalized applying beta actin and GAPDH as standards. Chemiluminescence signals were captured using the ChemiDoc XRS System. Variations in protein levels were determined by densitometric scanning and normalized to internal requirements. FDA and IRB permitted single patient clinical trial A 86 year old patient with high level androgen independent metastatic prostate cancer was agreed for the Natura alpha trial treatment for his condition with approval from the IRB and FDA. Natura leader was given orally with increasing amounts from 40mg, 80mg, 160 to 200 mg daily every two weeks and 200 mg down the road for 3 months.