In Vivo Immunomodulatory, Cumulative Skin Irritation, Sensitization and Effect of d-Limonene on Permeation of 6-Mercaptopurine through Transdermal Drug Delivery

• Published in: Biological & Pharmaceutical Bulletin Vol. 31; no. 4; pp. 656 - 661
• Main Authors: Chandrashekar, N. S., Hiremath, Shobha Rani Rajeev
• Format: Journal Article
• Language: English
• Published: Japan The Pharmaceutical Society of Japan 01.04.2008; Pharmaceutical Society of Japan; Japan Science and Technology Agency
• Subjects: 6-mercaptopurine; Administration, Cutaneous; Animals; Blood Cell Count; Bone Marrow Cells - drug effects; Cell Membrane - drug effects; Cell Membrane - metabolism; Chemical Phenomena; Chemistry, Physical; Cyclohexenes - adverse effects; Cyclohexenes - pharmacology; d-limonene; Diffusion Chambers, Culture; Humans; immunomodulatory; Immunosuppressive Agents - administration & dosage; Immunosuppressive Agents - adverse effects; Immunosuppressive Agents - pharmacology; In Vitro Techniques; Irritants - pharmacology; Limonene; Mercaptopurine - administration & dosage; Mercaptopurine - adverse effects; Mercaptopurine - pharmacology; Mice; Mice, Inbred BALB C; Skin - pathology; Skin Absorption - drug effects; Terpenes - adverse effects; Terpenes - pharmacology; transdermal
• ISSN: 0918-6158; 1347-5215
• DOI: 10.1248/bpb.31.656

Using skin as a port for systemic drug administration, transdermal drug delivery has expanded greatly over the last two decades. Our aim was to formulate the single layer drug-in-adhesive transdermal patch for 6-mercaptopurine (6-MP). In vitro permeation study was carried out using modified Franz diffusion cell with and without of different concentration of d-limonene in human cadaver skin. In vivo immunomodulatory was carried out in mice, cumulative skin irritation, sensitization and patch adherence study was done in both mice and human subjects. 6-MP flux increased from 43±12.2 μg/cm2h (control) to 162.8±32.2 μg/cm2h (6% w/v d-limonene) data was significant (p<0.05), with decrease in the lag time to 35±9.3 min compared to control of 90 ±15.3 min. In vivo immunomodulatory effect was shown in the Balb/c mice with 100 μmol/kg/body wt of animal for 5d (one dose/d) of d-limonene. WBC count of 13469 cells/mm peak was observed on 12th day, bone marrow cells of 26.3×106 cells/femur and α-esterase positive cells of 1259±328.4 cells/4000 bone marrow cells. Cumulative skin irritation, sensitisation and patch adherence in animals and human subjects showed no skin irritation and sensitization. Patch adhesion was greater than 90.0% respectively in both human subjects and mice. The percentage of human subjects with adhesive residue was significantly less with scores of zero. d-Limonene proved as good chemical enhancer by increasing in the skin permeability with shortened the lag time. It proved that therapeutic amount of 6-MP can be delivered through transdermal drug delivery.