Rubus creticus Root Extract and Its Active Fraction Reduce Calcium Oxalate Crystallization: An Antiurolithic-Guided Fractionation and In Silico ADMET Studies

Abstract

This study aimed to evaluate the anti-urolithiatic activities of Rubus creticus Tourn. ex L., Rosaceae, ethanol extract and its hexane, chloroform, ethyl acetate, and water fractions from the R. creticus root used as a kidney stone reducer in traditional medicine, and to reveal their effective compounds by phytochemical analysis. Anti-urolithiatic activity was investigated by calcium oxalate inhibition. Phytochemical content of active samples was investigated by GC-MS. Bioactivity-guided fractionation process was performed on the active Rubus chloroform extract using chromatographic methods. The in silico ADMET study was performed to predict the pharmacokinetic profile. Rubus chloroform extract exhibited a stronger anti-urolithiatic activity with an IC50 value of 9.94 & micro;g/ml compared to standard Cystone (R) (59.07 & micro;g/ml). It was revealed that the Rubus chloroform extract, which exhibited the best anti-urolithiatic activity, contained ethyl oleate, dehydroabietic acid, abietic acid, and hexadecanoic acid ethyl ester. It was observed that the isolate obtained as a single band in pTLC with anti-urolithiasis guidance from Rubus chloroform extract was a rich mixture of diterpene compounds (isopimaric acid, abietic acid, pimaric acid, kaur-16-en-19-al, and dehydroabietic acid) and this isolate (4.09 & micro;g/ml) also had significant activity. In silico ADMET evaluation of the diterpenes revealed high gastrointestinal absorption and favorable permeability. The results showed that Rubus chloroform extract exhibited strong anti-urolithiatic activity, likely due to its diterpenes and other contributing secondary metabolites. The findings suggest a scientific basis for the ethnobotanical use of the plant, showing that it may serve as a prospective candidate for developing natural remedies against kidney stone disease. However, in vivo studies are necessary to fully validate its efficacy.