Advertisement

AAPS PharmSciTech

, Volume 19, Issue 5, pp 2264–2275 | Cite as

Development of a Topical 48-H Release Formulation as an Anti-scarring Treatment for Deep Partial-Thickness Burns

  • Rossella Dorati
  • Jorge L. Medina
  • Patrick P. DeLuca
  • Kai P. Leung
Research Article

Abstract

The purpose of this study was to develop pirfenidone (PF) ointment formulations for a dose finding study in the prophylactic treatment of deep partial-thickness burns in a mouse model. A preformulation study was performed to evaluate the solubility of PF in buffers and different solvents and its stability. Three different formulations containing 1, 3.5, and 6.5% w/w PF were prepared and optimized for their composition for testing in mice. Optimized formulations showed promising in vitro release profiles, in which 20–45% of PF was released in the first 7 h and 70–90% released within 48 h. The rheological properties of the ointment remained stable throughout storage at 25 ± 2°C/60% RH. Animal studies showed treatments of burn wounds during the inflammatory stage of wound healing with PF ointments at different drug concentrations had no adverse effects on reepithelization. Moreover, 6.5% PF ointment (F3) reduced the expression of pro-inflammatory cytokines IL-12p70 and TNFα. This study suggests that hydrocarbon base ointment could be a promising dosage form for topical delivery of PF in treatment of deep partial-thickness burns.

KEY WORDS

pirfenidone ointment formulations burn treatment deep partial-thickness burn mice 

Notes

Funding Information

RD performed this study at the US Army Institute of Surgical Research during her sabbatical leave from the University of Pavia, Italy. Her stipend was supported by an appointment to the Postgraduate Research Participation Program at the US Army Institute of Surgical Research administered by the Oak Ridge Institute for Science and Education through an interagency agreement between the US Department of Energy and US Army Medical Research and Materiel Command (USAMRMC). This work was supported in part through the Congressionally Directed Medical Research Programs, USAMRMC W81XWH-15-2-0083, and the Naval Medical Research Center’s Advanced Medical Development program (MIPR N3239815MHX040).

Compliance with Ethical Standards

This study has been conducted in compliance with the Animal Welfare Act, the implementing Animal Welfare Regulations, and the principles of the Guide for the Care and Use of Laboratory Animals.

Conflict of Interest

The authors declare that they have no conflict of interest.

Disclosure

The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or as reflecting the views of the Department of the Army or the Department of Defense.

Supplementary material

12249_2018_1030_Fig12_ESM.gif (291 kb)
Supplemental Figure 1

Histological mouse skin sections 3 days after a deep partial-thickness burn treated with pirfenidone ointment. Images depict H&E stained sections from burn wounds 3 days after thermal injury and treated with A) placebo, B) 1%, C) 3.5%, and D) 6.5% pirfenidone ointment. Black arrow heads point to the burn wound edges showing a loss of the epidermis between. Insets are from the areas marked by the lines indicating the left burn wound edge. Left of the black arrows in the insets point at the leading edge of the intact epidermis. (GIF 291 kb)

12249_2018_1030_MOESM1_ESM.tif (4.2 mb)
High-resolution image (TIF 4274 kb)
12249_2018_1030_Fig13_ESM.gif (376 kb)
Supplemental Figure 2

Histological mouse skin sections 12 days after a deep partial-thickness burn treated with pirfenidone ointment. Images depict H&E stained sections from burn wounds 12 days after thermal injury and treated with A) placebo, B) 1%, C) 3.5%, and D) 6.5%pirfenidone ointment. Many of the wounds completely reepithelialized 12 days after the burn and most were close to full reepithelialization. Black arrow heads point at the left wound edges or wound centers. Insets are from the areas marked by the brackets. Left of the black arrows if present in the insets point at the leading edge of the intact epidermal layer. (GIF 375 kb)

12249_2018_1030_MOESM2_ESM.tif (7.8 mb)
High-resolution image (TIF 7969 kb)
12249_2018_1030_Fig14_ESM.gif (477 kb)
Supplemental Figure 3

Histological mouse skin sections 22 days after a deep partial-thickness burn treated with pirfenidone ointment. Images depict H&E stained sections from burn wounds 22 days after thermal injury and treated with A) placebo, B) 1%, C) 3.5%, and D) 6.5% pirfenidone ointment. Most burn wounds completely reepithelialized 22 days after the burn injury. (GIF 477 kb)

12249_2018_1030_MOESM3_ESM.tif (6.8 mb)
High-resolution image (TIF 6997 kb)

References

  1. 1.
    Tredget EE, Levi B, Donelan MB. Biology and principles of scar management and burn reconstruction. Surg Clin North Am. 2014;94(4):793–815.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Qian L-W, Fourcaudot AB, Yamane K, You T, Chan RK, Leung KP. Exacerbated and prolonged inflammation impairs wound healing and increases scarring. Wound Repair Regen. 2016;24(1):26–34.CrossRefPubMedGoogle Scholar
  3. 3.
    Penn JW, Grobbelaar AO, Rolfe KJ. The role of the TGF-β family in wound healing, burns and scarring: a review. Int J Burns Trauma. 2012;2(1):18–28.PubMedPubMedCentralGoogle Scholar
  4. 4.
    Karagoz H, Sever C, Bayram Y, Sahin C, Kulahci Y, Ulkur E. A review of the prevention and treatment of hypertrophic scars: part I clinical aspects. Arch Clin Exp Surg (ACES). 2012;1(4):237–48.CrossRefGoogle Scholar
  5. 5.
    Aarabi S, Longaker MT, Gurtner GC. Hypertrophic scar formation following burns and trauma: new approaches to treatment. PLoS Med. 2007;4(9):1464–70.CrossRefGoogle Scholar
  6. 6.
    Wasiak J, Cleland H, Campbell F, Spinks A. Dressings for superficial and partial thickness burns. Cochrane Database Syst Rev. 2013;3:1–85.Google Scholar
  7. 7.
    J Schaefer C, W Ruhrmund D, Pan L, Seiwert S, Kossen K. Antifibrotic activities of pirfenidone in animal models. Eur Respir Rev. 2011;20(120):85–97.Google Scholar
  8. 8.
    Veras-Castillo ER, Cardenas-Camarena L, Lyra-Gonzalez I, Munoz-Valle JF, Lucano-Landeros S, Guerrerosantos J, et al. Controlled clinical trial with pirfenidone in the treatment of breast capsular contracture: association of TGF-beta polymorphisms. Ann Plast Surg. 2013;70(1):16–22.CrossRefPubMedGoogle Scholar
  9. 9.
    Armendariz-Borunda J, Lyra-Gonzalez I, Medina-Preciado D, Gonzalez-Garcia I, Martinez-Fong D, Miranda RA, et al. A controlled clinical trial with pirfenidone in the treatment of pathological skin scarring caused by burns in pediatric patients. Ann Plast Surg. 2012;68(1):22–8.CrossRefPubMedGoogle Scholar
  10. 10.
    NIH U.S. National Library of Medicine - ClinicalTrials.gov [cited 12.14.2017]. Available from: https://clinicaltrials.gov/ct2/results?term=pirfenidone+AND+fibrosis&Search=Search.
  11. 11.
    Rowan MP, Cancio LC, Elster EA, Burmeister DM, Rose LF, Natesan S, et al. Burn wound healing and treatment: review and advancements. Crit Care. 2015;19:1–12.CrossRefGoogle Scholar
  12. 12.
    Hall CL, Wells AR, Leung KP. Pirfenidone reduces profibrotic responses in human dermal myofibroblasts, in vitro. Lab Investig. 2018.  https://doi.org/10.1038/s41374-017-0014-3
  13. 13.
    Moran N. p38 kinase inhibitor approved for idiopathic pulmonary fibrosis. Nat Biotech. 2011;29(4):301.CrossRefGoogle Scholar
  14. 14.
    Furukawa F, Matsuzaki K, Mori S, Tahashi Y, Yoshida K, Sugano Y, et al. p38 MAPK mediates fibrogenic signal through smad3 phosphorylation in rat myofibroblasts. Hepatology. 2003;38(4):879–89.Google Scholar
  15. 15.
    Rodríguez-Castellanos M, Tlacuilo-Parra A, Sánchez-Enríquez S, Vélez-Gómez E, Guevara-Gutiérrez E. Pirfenidone gel in patients with localized scleroderma: a phase II study. Arthritis Res Ther. 2014;16(510):1–8.Google Scholar
  16. 16.
    Giri SN, Margolin SB. Effects of topical application of pirfenidone ointment on thermoplasty-induced acute lameness in a double-blind and acute and chronic lameness of musculoskeletal origin in an open multi-centered field trial in horses. Res Commun Mol Pathol Pharmacol. 2005;117-118:47–63.PubMedGoogle Scholar
  17. 17.
    Parmar V, Desai SB, Vaja T. RP-HPLC and UV spectrophotometric methods for estimation of pirfenidone in pharmaceutical formulations. Indian J Pharm Sci. 2014;76:225–30.Google Scholar
  18. 18.
    Thorat S, P Padmane S, R Tajne M, M Ittadwar A. Development and validation of simple, rapid and sensitive UV, HPLC and HPTLC methods for the estimation of pirfenidone in tablet dosage form. J Chil Chem Soc. 2016;61(2):2978–81.Google Scholar
  19. 19.
    Babu Bodempudi S, Babur R, Srinivasa Reddy K. Development and substantiation of a RP-HPLC method for monitoring of impurities in pirfenidone drug substance. Am J Anal Chem. 2015;06:1019–29.Google Scholar
  20. 20.
    Salerno C, Carlucci AM, Bregni C. Study of in vitro drug release and percutaneous absorption of fluconazole from topical dosage forms. AAPS PharmSciTech. 2010;11(2):986–92.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Lokhandwala H, Deshpande A, Deshpande S. Kinetic modeling and dissolution profiles comparison: an overview. Int J Pharm. 2013;4(1):728–38.Google Scholar
  22. 22.
    Rajesh Asija SP, Sangeeta Asija, Sandeep Kataria Formulation and evaluation of herbal ointment of Murraya koenigii. Int J Pharm Res Bio-Sci 2015;4(3):433–443.Google Scholar
  23. 23.
    Said J, Dodoo CC, Walker M, Parsons D, Stapleton P, Beezer AE, et al. An in vitro test of the efficacy of silver-containing wound dressings against Staphylococcus aureus and Pseudomonas aeruginosa in simulated wound fluid. Int J Pharm. 2014;462(1):123–8.Google Scholar
  24. 24.
    ICH Topic Q 1 A (R2) Stability testing of new drug substances and products. Note for guidance on stability testing: stability testing of new drug substances and products (CPMP/ICH/2736/99) [Internet]. [cited 12.14.2017]. Available from: http://www.ich.org/products/guidelines/quality/quality-single/article/stability-testing-of-new-drug-substances-and-products.html.
  25. 25.
    Medina JL, FFA, Sebastian EA, Shankar R., Brown AW and, KP L Standardization of deep partial-thickness scald burns in C57BL/6 mice. Int J Burn Trauma. 2018;(8)2Google Scholar
  26. 26.
    Zheng X-F, Hong Y-X, Feng G-J, Zhang G-F, Rogers H, Lewis MAO, et al. Lipopolysaccharide-induced M2 to M1 macrophage transformation for IL-12p70 production is blocked by Candida albicans mediated up-regulation of EBI3 expression. PLoS One. 2013;8(5):1–10.Google Scholar
  27. 27.
    Ashcroft GS, Jeong M-J, Ashworth JJ, Hardman M, Jin W, Moutsopoulos N, et al. Tumor necrosis factor-alpha (TNF-α) is a therapeutic target for impaired cutaneous wound healing. Wound Repair Regen. 2012;20(1):38–49.Google Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2018

Authors and Affiliations

  • Rossella Dorati
    • 1
    • 2
  • Jorge L. Medina
    • 1
  • Patrick P. DeLuca
    • 3
  • Kai P. Leung
    • 1
  1. 1.Dental and Craniofacial Trauma Research and Tissue RegenerationUS Army Institute of Surgical ResearchHoustonUSA
  2. 2.Department of Drug SciencesUniversity of PaviaPaviaItaly
  3. 3.College of PharmacyUniversity of KentuckyLexingtonUSA

Personalised recommendations