| Summary: | <i>Pistacia lentiscus</i> L. has traditionally been employed as a diuretic and stimulant in the treatment of hypertension. Our interest centered on analyzing the chemical profile of the plant’s leaves and its in vitro, in vivo, and in silico antioxidant, antimicrobial, anticoagulant, and antidiabetic effects in order to valorize this species and prepare new high-value products that can be used in the agro-food and pharmaceutical industries. When this species’ essential oil was hydrodistilled and subjected to GC-MS analysis, the results showed that the principal components were germacrene D (17.54%), spathulenol (17.38%), bicyclogermacrene (12.52%), and terpinen-4-ol (9.95%). The extraction of phenolic compounds was carried out by decoction and Soxhlet. The determination of total polyphenols, flavonoids, and tannins of aqueous and organic extracts by spectrophotometric methods demonstrated the richness of this species in phenolic compounds. Chromatographic analysis by HPLC/UV-ESI-MS of the aqueous extract of <i>P. lentiscus</i> revealed the presence of 3,5-di-<i>O</i>-galloyl quinic acid, gallic acid, and 3,4,5-tri-<i>O</i>-galloyl quinic acid specific to this species. The study of antioxidant activity by three methods (DPPH, FRAP, and Total Antioxidant Capacity) revealed that <i>P. lentiscus</i> is a very promising source of natural antioxidants. The antimicrobial activity of the essential oil and aqueous extract (E<sub>0</sub>) was studied by microdilution on the microplate. The results revealed the effectiveness of the aqueous extract compared to the essential oil against Gram-negative bacteria (<i>K. pneumoniae</i>, <i>A. baumannii</i>, <i>E. aerogenes</i>, <i>E. cloacae</i>, <i>P. fluorescence</i>, <i>Salmonella</i> sp., <i>Shigella</i> sp., and <i>Y. enterolitica</i>) and candidoses (<i>C. krusei</i> and <i>C. albicans</i>). The measurements of prothrombin time (PT) and activated partial thromboplastin time (aPTT) of the aqueous extract (E<sub>0</sub>) can significantly prolong these tests from concentrations of 2.875 and 5.750 mg/mL, respectively. The antihyperglycemic effect of the aqueous extract (E<sub>0</sub>) showed a strong in vitro inhibitory activity of α-amylase and α-glucosidase compared to acarbose. Thus, it significantly inhibited postprandial hyperglycemia in Wistar albino rats. The in-silico study of the major compounds of the essential oil and extract (E<sub>0</sub>) carried out using PASS, SwissADME, pkCSM, and molecular docking tools confirmed our in vitro and in vivo results. The studied compounds showed a strong ability to be absorbed by the gastrointestinal tract and to passively diffuse through the blood-brain barrier, a similarity to drugs, and water solubility. Molecular docking experiments deduced the probable mode of action of the identified compounds on their respective target proteins, such as NADPH oxidase, thrombin, α-amylase, and α-glucosidase. Furthermore, given the demonstrated antioxidant, antimicrobial, anticoagulant, and antidiabetic effects, we can affirm the richness of <i>P. lentiscus</i> in bioactive molecules and its use in traditional medicine as a source of preservative agent.
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