Lactuca orientalis (Boiss.) Boiss. is one of the most frequently used ethnomedicinal plant. This research study was designed to decipher the phytochemical screening, pharmacological potential and implementation of scanning electron microscope (SEM). Six different solvents were used to prepare L. orientalis (LO) seed extracts. Phytochemical and antioxidant activities were determined calorimetrically. To investigate antidiabetic, α-amylase inhibition assay was performed. Brine shrimp assay was performed for cytotoxicity and anti-leishmanial via MTT assay. Disc-diffusion assay was performed to detect protein kinase inhibitory, antibacterial and antifungal activities. SEM was used as identification tool. Significant amount of phenolic and flavonoid content were identified in methanol extract (LOSM) (95.76 ± 3.71 GAE/mg) and (77 ± 3.60 QE/mg). Highest DPPH scavenging potential (82%) was reported for LOSM. Significant total antioxidant capacity (90.60 ± 1.55 AAE/mg) and total reducing power (94.44 ± 1.38 AAE/mg) were determined for LOSM. Highest α-amylase inhibition was found in LOSM (78.20 ± 1.58%). The highest LD50 of brine shrimp was found for n-Hexane extract (LOSH) 13.03 𝜇g/ml. All extracts showed strong anti-leishmanial activity except LOSH. L. orientalis seeds showed significant protein kinase inhibition, antibacterial and antifungal activities. The seeds of L. orientalis were seen to be oblong to obovate, projections, wavy slightly straight, anticlinal wall was raised with apex acuminate. The outer-periclinal wall convex with fine texture. In conclusion, our findings scientifically support ethnomedicinal and biological potentials of L. orientalis seeds. In future, L. orientalis seeds need to be explored for identification and isolation of bioactive compounds. The results obtained necessitate further in vivo studies to evaluate their pharmacological potentials.