The study investigated prolonged drought stress effects on phytohormone and polyamine levels in cress (Lepidium sativum L.) leaves, elucidating plant responses to harsh environments. Phytohormones, 12-oxophytodienoic acid (OPDA), Jasmonic acid (JA), Jasmonyl-Isoleucine (JA-Ile), Abscisic acid (ABA), and salicylic acid (SA) increased, especially ABA that rose consistently. Indole-3-acetic acid (IAA) responded akin to ABA and SA. Cytokinins such as zeatin (ZT) and isopentenyl adenine (iP) diversified drought tolerance, ZT fell sharply, and iP remained stable. Gibberellic acid (GA3) declined, while brassinolide (BL) surged with prolonged drought. Polyamines reduced, indicating sensitivity to severe drought. Findings uncovered intricate regulatory mechanisms for plant acclimation to water scarcity. Interaction of OPDA, JA, JA-Ile, ABA, SA, IAA, ZT, iP, GA3, BL, and polyamines contributes to drought coping and physiological balance. Insights into cress leaf response and drought tolerance mechanisms contribute significantly to plant drought acclimation understanding.