<p>This study aimed to evaluate the anti-urolithiatic activities of <i>Rubus creticus</i> Tourn. ex L., Rosaceae, ethanol extract and its hexane, chloroform, ethyl acetate, and water fractions from the <i>R. creticus</i> 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 <i>Rubus</i> chloroform extract using chromatographic methods. The <i>in silico </i>ADMET study was performed to predict the pharmacokinetic profile. <i>Rubus</i> chloroform extract exhibited a stronger anti-urolithiatic activity with an IC₅₀ value of 9.94&#xa0;µg/ml compared to standard Cystone® (59.07&#xa0;µg/ml). It was revealed that the <i>Rubus</i> 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 <i>Rubus</i> 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&#xa0;µg/ml) also had significant activity. <i>In silico </i>ADMET evaluation of the diterpenes revealed high gastrointestinal absorption and favorable permeability. The results showed that <i>Rubus</i> 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, <i>in vivo</i> studies are necessary to fully validate its efficacy.</p> Graphical Abstract <p></p>