Determination of ursolic acid in ethanolic extract of Rotheca serrata (L.) Steane & Mabb. by HPLC method.

Rotheca serrata (L.) Steane & Mabb. a plant having significant medicinal value is used in treating a fever, rheumatism, inflammation, pain, and malarial fever. Ursolic acid has a five-ringed pentacyclic structure belongs to the family of triterpenoids. It has antioxidant, antitumor, and hepato-protective properties. The method in HPLC was developed on a SHIMADZU, Shimpack ODS (25 cm × 4.6 mm, 5 µm) column. The mobile phase consisted of a 70:25:5 (v/v/v) mixture of methanol, acetonitrile, and 0.02 M ammonium acetate buffer (pH 3.5). A flow rate of 1.2 mL/min and λmax of 215 nm. Ursolic acid from the extract and standard was eluted at the same retention time of 4.1 min. 20-120 µg/mL was the linearity range chosen and was found to be linear (R2=0.996). The value of LOD was 0.85 µg/mL and LOQ was 1.96 µg/mL. The method for analysis of ursolic was found to be accurate with good repeatability.

Combinatorial Implications of Nrf2 Inhibitors with FN3K Inhibitor: In vitro Breast Cancer Study.

BACKGROUND: Platinum derivatives are chemotherapeutic agents preferred for the treatment of cancers including breast cancer. Oxaliplatin is an anticancer drug that is in phase II studies to treat metastatic breast cancer. However, its usage is constrained by chemoresistance and dose-related side effects. OBJECTIVE: The objective of this study is to examine the combinatorial efficacy of brusatol, an Nrf2 blocker, with oxaliplatin (a proven FN3K blocker in our study) in mitigating breast cancer growth in vitro. METHODS: We performed cytotoxicity assays, combination index (CI) analysis, colony formation assays, apoptosis assays, and Western blotting. RESULTS: Results of our study described the chemosensitizing efficacy of brusatol in combination with lowdose oxaliplatin against breast cancer through synergistic effects in both BT-474 and T47D cells. A significant mitigation in the migration rate of these cancer cells was observed with the combination regimen, which is equivalent to the IC-50 dose of oxaliplatin (125 µM). Furthermore, ROS-mediated and apoptotic modes of cell death were observed with a combinatorial regimen. Colony formation of breast cancer cell lines was mitigated with a combinatorial regimen of bursatol and oxaliplatin than the individual treatment regimen. FN3K expression downregulated with oxaliplatin in T47D cells. The mitigation of FN3K protein expression with a combination regimen was not observed but the Nrf2 downstream antioxidant signaling proteins were significantly downregulated with a combination regimen similar to individual drug regimens. CONCLUSION: Our study concluded the combination efficacy of phytochemicals like brusatol in combination with low-dose oxaliplatin (FN3K blocker), which could enhance the chemosensitizing effect in breast cancer and minimize the overall dose requirement of oxaliplatin.