5-Benzyl-1,3,4-oxadiazole-2-thiol (OXPA), synthesized as a series of active compounds, has not been investigated extensively, despite possessing a pharmacophore, known for a number of pharmacological properties. Therefore, the present study aimed to investigate the compound for drug qualifying properties, develop analytical methods and perform biological screening for antidiabetic, antioxidant, antibacterial, anti-TB, anti-inflammatory and antiangiogenic activities. The compound was evaluated for drug-likeliness using a number of computational software. Keeping in view the presence of a UV absorbing chromophore, a UV spectrophotometric method was developed and validated at 264 nm for determining the compound in bulk and stress solutions.For more specific and stability indicating assay, RP-HPLC methods with diode array detection (DAD) were also developed and validated to determine the compound in bulk, stress solutions and rat plasma. Afterwards, the compound was subjected to antibacterial activity studies against Gram-positive, Gram-negative, H. pylori and Mycobacterium tuberculosis (H37 Rv) strains and clinical isolates. Anti-inflammatory activity was determined using protein denaturation, anti-proteinase, membrane stabilization assays, and rat-paw edema model. Antiangiogenic activity was determined using the CAM assay. Finally, the pharmacokinetics parameters were determined in rats following oral administration of the compound. Molecular and physicochemical parameter, bioactivity and toxicity, determined computationally, indicated that the compound passed the drug-like filters and qualify drug-likeliness. The compound was expected to have promising xxvii antidiabetic, antioxidant, antibacterial, anti-inflammatory and antiangiogenic activities, low toxicity and good oral absorption. The UV spectrophotometric method developed and validated at 264 nm was found to be linear (0.25-40.00 µg/mL, R2= 0.9984), sensitive (LOD = 0.109 µg/ml and LOQ = 0.332 µg/ml), specific, accurate, precise and robust. Reversed-phase, isocratic elution of the compound using isocratic mobile phase (ammonium acetate buffer (0.1%): acetonitrile, 70:30, V/V), at a flow rate of 1 mL/min produced Gaussian peak fulfilling all the system suitability parameters. Likewise, the fulfilled all the method validation ICH guidelines; recovery (96.27-100.44%), intraday accuracy and precision (97.20-99.47%, RSD < 5) and inter-day accuracy precision (97.59-98.15%, RSD < 5%). Furthermore, the method was stability indicating because the determination was not affected by forced-degradation products in the presence of different stressors. In mild to severe stress conditions, compound degraded to variable extent in acidic and basic hydrolysis and in oxidative stress (30% H2O2). The analytical sample remained stable throughout the study period in refrigerator and in three freeze thaw cycles. HPLC method for determination of the compound in plasma indicated that peak of the compound was not affected by plasma impurities and degradation products. Moreover, the method fulfilled the ICH method validation guidelines; recovery (94.15-101.88%), intraday accuracy and precision (100.08-114.14%, RSD < 15%) and inter-day accuracy precision (100.4-114.8%, RSD < 15%). The compound showed antidiabetic activity comparable to the standards in the glucose uptake by yeast cells, inhibition of hemoglobin glycosylation and alpha xxviii amylase assays. The compound exhibited good interaction with antidiabetic enzymes. Antioxidant activity of the compound was comparable to vitamin C in DPPH and lipid peroxidation assays (P < 0.05). Moreover, it preserved and protected the antioxidant status and liver of rats against induced-oxidative stress. The compound showed promising antibacterial activity against Bacillus subtilis and Escherichia coli (MIC=62.5 µg/mL) and Bacillus pumilus, Pseudomonas aeruginosa, Salmonella enterica, H-pylori (MIC = 125 µg/mL) and rifampicin resistant and standard mycobacterium strains (MIC = 40 µg/mL). The compound showed anti-inflammatory activity in different models as protein denaturation (47.02 ± 0.55%), anti-proteinase (64.30±1.88) and RBC hemolysis (35.78±1.1%). The compound also showed antiangiogenic effect in a dose dependent manner. Pharmacokinetics studies indicated that the compound achieved maximum concentration (32.19 µg/mL) at 2.09 h with area under the curve AUC 0-∞ (239.14 µg/mL*h). The results of the present study indicate that OXPA qualifies drug-like properties and has good antidiabetic and anti-inflammatory activities. Moreover, the methods developed for determination of OXPA are simple, sensitive and reliable, hence, may be used for determination of the compound in bulk and different matrices at sub-microgram level.