Mitragynine is a natural compound found in the leaves of the mitragyna speciosa tree, commonly known as kratom, which is primarily sourced from Southeast Asia. Mitragynine and its derivatives possess various pharmacological properties, including pain relief, anticancer effects, anti-inflammatory properties, anxiety reduction, stimulant properties, antidepressant effects, anti-addictive properties, antioxidant properties, and potential benefits for diabetes management. This review article highlights the methodologies of extraction techniques for isolating mitragynine, purification, characterization, biosynthesis, including the complete synthesis of mitragynine and its derivatives, and briefly summarizes their biological activities and toxicology of mitragynine. It is important to acknowledge that ongoing research is being conducted to explore the pharmacological effects of mitragynine, and further investigation is necessary to fully comprehend its impacts and potential medical applications. Moreover, the utilization of mitragynine and kratom remains contentious, with legal limitations imposed in numerous countries.