AbstractProtein‐ and peptide‐based therapies have emerged as a possible alternative to antibody‐ and small molecule‐ based therapies for treating cancer, aiming to improve drug localization and enhance effectiveness. Peptides such as cyclotides and acyclotides are known for their exceptional stability, which is attributed to the presence of a cystine knot. These peptides exhibit a diverse array of biological activities, making them promising candidates for therapeutic development. Transcriptomics approaches are frequently employed to gain insights into gene organization and identify variations in transcript expression levels. Furthermore, proteomics and NMR spectroscopy techniques are highly regarded as a potent method for analyzing peptide structures. This review offers a thorough examination of the most recent advancements and developments in the methods employed to isolate and characterize the peptides, including purification, proteomics, and transcriptomics. Furthermore, the article thoroughly analyzes the unique structural features of cyclotides and acyclotides, providing valuable insights into their potential groundbreaking applications. The discussion also explores the issuance of numerous patents in the medical and pharmaceutical field, highlighting the growing interest and potential applications of cyclotides in cancer treatment.