Wound healing stays a substantial health affair for a great number of patients globally. Hypoxia plays a decisive role in specifying the normal successfulness of healing process. Hypoxia-inducible factor-1alpha, plays a critical role in oxygen homeostasis, cell survival and migration and growth factor release throughout the healing process. The target of this research is to characterize the phytochemical and antioxidant analysis of the lipophilic extract of Calendula officinalis (C. officinalis) flowers utilizing colorimetric, spectrophotometric, and chromatographic techniques. In addition, studying the wound healing properties in vivo and in vitro model. Rats were divided into four groups (six in each group) as follow: Group I: Control rats. Group II: Wounded and untreated rats. Group III: Rats are treated topically with 2.5% of C. officinalis in gel. Group IV Rats are treated topically with 5% of C. officinalis in gel. Furthermore, Molecular docking study was conducted to elucidate the mechanistic of C. officinalis wound healing properties through the inhibitory effects of its main components against key anti-healing goals and study the best binding affinity. Using GC-MS analysis six main different organic classes fatty acid (36.99 %), Sesquiterpene (34.9%), hydrocarbons (20.06%), diterpene (3.9%), phytosterol (2.10%) and alkaloid (0.53%), and 24 components comprising 98.48% of the observed peaks were identified as a consequence. It is evident that the predominant fatty acid is 1-heptatriacotanol (16.47%). Additionally, Isochiapin B is a significant sesquiterpene compounds that accounting for 15.91% of all sesquiterpene compounds. Moreover, this extract exhibited more potent antioxidant activity were observed., as demonstrated by the IC50. Owing to its high concentration of active compounds, C. officinalis promoted more wound contraction., collagen synthesis, 5' adenosine monophosphate-activated protein kinase (AMPK), Hypoxia-inducible factor 1-alpha (HIF-1α) and accelerated epithelization with a decrease in matrix metalloproteinases (MMP-9). Molecular docking revealed that Isochiapin B had better binding affinities than 1-Heptatriacotanol in the interaction with AMPK, Collagenase, HIF-1α, MMP-9, tumor necrosis factor alpha (TNF-α), and vascular endothelial growth factor (VEGF), and that its best binding affinity was with MMP-9. Obtained results assured that Calendula officinalis L possesses considerable wound healing activity in wounded rats due to its potent antioxidant and anti-inflammatory effects.