Local and Systemic Effects of Topical Betulinic Acid in a Psoriasis-like Inflammation Model in Mice

Abstract

Psoriasis patients often discontinue oral and injectable treatments due to concerns about both safety and efficacy. Issues such as adverse side effects, limited long-term effectiveness, and fear of potential complications contribute to non-adherence, impacting treatment outcomes and patientsʼ quality of life. Betulinic acid (BA) forms supramolecular aggregates through self-assembly in a hydroalcoholic vehicle, which was hypothesized to have antipsoriatic activity when applied topically. To test this, imiquimod was applied to induce psoriasis-like skin inflammation in mice (except in the untreated group) every 24 hours for 5 days. Two hours after each imiquimod application, the groups received either 100 µL of vehicle (10% glycerol aqueous solution), 0.05 mg/mL clobetasol (Clo), or 0.5 mg/mL BA. At the end of the study, the Psoriasis Area and Severity Index (PASI) was evaluated (n = 12/group), and complete skin clearance time (CSC) was determined in six mice per group. The remaining six mice per group were used to assess acanthosis, lymphocyte and granulocyte infiltration, and Akt and ERK phosphorylation in skin samples, as well as TNFα, IL-17A, IFNγ, and TGFβ levels in serum. To assess treatment safety, we evaluated food and water intake, ambulation pattern, body weight gain, organ weights, and blood parameters. BA significantly reduced CSC time by up to 40% compared to the control and was 10% faster than Clo. Both BA and Clo reduced PASI and acanthosis to approximately one-third of control values, normalized immune cell infiltration and TNFα levels, decreased IL-17A by more than 30%, and reduced p-Akt and p-ERK2. BA uniquely normalized IFNγ levels without causing intolerable toxicity. Using an animal model of psoriatic skin inflammation, our findings support BA as a strong candidate for clinical translation, warranting further studies on its safety, pharmacokinetics, and optimal dosage in humans, potentially leading to randomized controlled trials in psoriasis patients.

Keywords

Dillenia indica - Dilleniaceae - betulinic acid - topical application - psoriasis-like skin inflammation - efficacy-safety

Publikationsverlauf

Eingereicht: 21. Oktober 2024

Angenommen nach Revision: 27. Juni 2025

Accepted Manuscript online:
27. Juni 2025

Artikel online veröffentlicht:
05. August 2025

• References

• 1 World Health Organization (WHO). Global Report on Psoriasis. Geneva: Who Press; 2016: 5-6 Zugriff am 23. Juni 2025 unter: https://www.who.int/publications/i/item/9789241565189
• 2 Lebwohl MG, Bachelez H, Barker J, Girolomoni G, Kavanaugh A, Langley RG, Paul CF, Puig L, Reich K, van der Kerkhof PCM. Patient perspectives in the management of psoriasis: Results from the population-based multinational assessment of psoriasis and psoriatic arthritis survey. J Am Acad Dermatol 2014; 70: 871-881
• 3 Lee HJ, Kim M. Challenges and future trends in the treatment of psoriasis. Int J Mol Sci 2023; 24: 13313
• 4 Bildziukevich U, Özdemir Z, Wimmer Z. Recent achievements in medicinal and supramolecular chemistry of betulinic acid and its derivatives. Molecules 2019; 24: 3546
• 5 Hordyjewska A, Ostapiuk A, Horecka A, Kurzepa J. Betulin and betulinic acid: Triterpenoids derivatives with a powerful biological potential. Phytochem Rev 2019; 18: 929-951
• 6 Gheorgheosu D, Duicu O, Dehelean C, Soica C, Muntean D. Betulinic acid as a potent and complex antitumor phytochemical: A minireview. Anticancer Agents Med Chem 2014; 14: 936-945
• 7 Kviecinski MR, David IMB, Fernandes FS, Correa MR, Clarinda MM, Freitas AF, Silva J, Gava M, Müller SD, Florentino D, Petronilho F, Mortele D, Kanis LA, Pedrosa RC. Healing effect of Dillenia indica fruit extracts standardized to betulinic acid on ultraviolet radiation-induced psoriasis-like wounds in rats. Pharm Biol 2017; 55: 641-648
• 8 Fernandes FS, da Silva GS, Hilel AS, Carvalho AC, Remor KVT, Schlindwein AD, Kanis LA, Martins DF, Kviecinski MR. Study of the potential adverse effects caused by the dermal application of Dillenia indica L. fruit extract standardized to betulinic acid in rodents. PLoS One 2019; 14: e0217718
• 9 Liu C, Chen Y, Lu C, Chen H, Deng J, Yan Y, Xu YY, Liu H, Huang H, Wei J, Han L, Dai Z. Betulinic acid suppresses Th17 response and ameliorates psoriasis-like murine skin inflammation. Int Immunopharmacol 2019; 73: 343-352
• 10 Del Rosso JQ. Topical corticosteroid therapy for psoriasis-a review of clobetasol propionate 0.025 % cream and the clinical relevance of penetration modification. J Clin Aesthet Dermatol 2020; 13: 22-29
• 11 van der Fits L, Mourits S, Voerman JSA, Kant M, Boom L, Laman JD, Cornelissen F, Mus AM, Florencia E, Prens EP, Lubberts E. Imiquimod-induced psoriasis-like skin inflammation in mice is mediated via the IL-23/IL-17 Axis. J Immunol 2009; 182: 5836-5845
• 12 Parmar KM, Itankar PR, Joshi A, Prasad SK. Anti-psoriatic potential of Solanum xanthocarpum stem in Imiquimod-induced psoriatic mice model. J Ethnopharmacol 2017; 198: 158-166
• 13 Langley RG, Ellis CN. Evaluating psoriasis with psoriasis area and severity index, psoriasis global assessment, and lattice system physicianʼs global assessment. J Am Acad Dermatol 2004; 51: 563-569
• 14 Hawkes JE, Gudjonsson JE, Ward NL. The snowballing literature on imiquimod-induced skin inflammation in mice: A critical appraisal. J Invest Dermatol 2017; 137: 546-549
• 15 Nestle OF, Kaplan DH, Barker J. Mechanisms of disease: Psoriasis. N Engl J Med 2009; 361: 496-509
• 16 Reich K, Iversen L, Bachelez H. Is complete skin clearance in psoriasis the answer?. European Medical Journal 2018; 6: 53-61
• 17 Daniel W, Cross C. Biostatistics: A Foundation for Analysis in Health Sciences. 11. Aufl.. New York: John Wiley & Sons; 2018
• 18 Huang SJ, Hijnen D, Murphy GF, Kuper TS, Calarese AW, Mollet IG, Schanbacher CF, Miller DM, Schmults CD, Clark RA. Imiquimod enhances IFN-γ production and effector function of T cells infiltrating human squamous cell carcinomas of the skin. J Invest Dermatol 2009; 129: 2676-2685
• 19 Reich K, Nestle FO, Papp K, Ortonne JP, Evans R, Guzzo C, Li S, Dooley LT, Griffiths CEM. EXPRESS study investigators. Infliximab induction and maintenance therapy for moderate-to-severe psoriasis: A phase III, multicentre, double-blind trial. Lancet 2005; 366: 1367-1374
• 20 He J, Zhao M, Ma X, Li D, Kong J, Yang F. The role and application of three IFN-related reactions in psoriasis. Biomed Pharmacother 2023; 167: 115603
• 21 Krzemień I, Szatan A, Skalska P, Cieślik M, Domagała A, Gębicka M, Bryniarski K, Nazimek K. Serum cytokine concentrations correlate with the severity of imiquimod and IL-23-induced psoriatic inflammation in mice. AIMS Allergy Immunol 2024; 8: 265-282
• 22 Di Meglio P, Nestle FO. The role of IL-23 in the immunopathogenesis of psoriasis. F1000 Biol Rep 2010; 2: 40
• 23 Ha HL, Wang H, Pisitkun P, Kim JC, Tassi I, Tang W, Morasso MI, Udey MC, Siebenlist U. IL-17 drives psoriatic inflammation via distinct, target cell-specific mechanisms. Proc Natl Acad Sci U S A 2014; 111: E3422-E3431
• 24 Knochelmann HM, Dwyer CJ, Bailey SR, Amaya SM, Elston DM, Mazza-McCrann JM, Paulos CM. When worlds collide: Th17 and Treg cells in cancer and autoimmunity. Cell Mol Immunol 2018; 15: 458-469
• 25 Han G, Williams CA, Salter K, Garl PJ, Li AG, Wang XJ. A role for TGF beta signaling in the pathogenesis of psoriasis. J Invest Dermatol 2010; 130: 371-377
• 26 Bettelli E, Carrier Y, Gao W, Korn T, Strom TB, Oukka M, Weiner HL, Kuchroo VK. Reciprocal developmental pathways for the generation of pathogenic effector TH17 and regulatory T cells. Nature 2006; 441: 235-238
• 27 Mahil SK, Capon F, Barker JN. Update on psoriasis immunopathogenesis and targeted immunotherapy. Semin Immunopathol 2016; 38: 11-27
• 28 El-Khattouti A, Selimovic D, Hannig M, Taylor EB, Abd Elmageed ZY, Hassan SY, Haikel Y, Kandil E, Leverkus M, Brodell RT, Megahed M, Hassan M. Imiquimod-induced apoptosis of melanoma cells is mediated by ER stress-dependent Noxa induction and enhanced by NF-κB inhibition. J Cell Mol Med 2016; 20: 266-286
• 29 Chamcheu JC, Adhami VM, Esnault S, Sechi M, Siddiqui IA, Satyshur KA, Syed DN, Dodwad SJM, Chavez-Rodriquez MI, Longley BJ, Wood GS, Mukhtar H. Dual Inhibition of PI3K/Akt and mTOR by the dietary antioxidant, delphinidin, ameliorates psoriatic features in vitro and in an imiquimod-induced psoriasis-like disease in mice. Antioxid Redox Signal 2017; 26: 49-69
• 30 Tothova C, Nagy O, Kovac G. Serum proteins and their diagnostic utility in veterinary medicine: A review. Vet Med (Praha) 2016; 61: 475-496
• 31 Yuan B, Wang X, Wang Z, Wei J, Qing C, Lu S. Comparison of fibrogenesis caused by dermal and adipose tissue injury in an experimental model. Wound Repair Regen 2010; 18: 202-210
• 32 Fraki JE, Briggaman RA, Lazarus GS. Transplantation of psoriatic skin onto nude mice. J Invest Dermatol 1983; 80: 31-35s
• 33 Zhan XK, Li JL, Zhang S, Xing PY, Xia MF. Betulinic acid exerts potent antitumor effects on paclitaxel-resistant human lung carcinoma cells (H460) via G2/M phase cell cycle arrest and induction of mitochondrial apoptosis. Oncol Lett 2018; 16: 3628-3634
• 34 Selzer E, Pimentel E, Wacheck V, Schlegel W, Pehamberger H, Jansen B, Kodym R. Effects of betulinic acid alone and in combination with irradiation in human melanoma cells. J Invest Dermatol 2000; 114: 935-940
• 35 Selzer E, Thallinger C, Hoeller C, Oberkleiner P, Wacheck V, Pehamberger H, Jansen B. Betulinic acid-induced Mcl-1 expression in human melanoma-mode of action and functional significance. Mol Med 2002; 8: 877-884
• 36 Pavlova NI, Savinova OV, Nikolaeva SN, Boreko EI, Flekhter OB. Antiviral activity of betulin, betulinic and betulonic acids against some enveloped and non-enveloped viruses. Fitoterapia 2003; 74: 489-492
• 37 Aiken C, Chen CH. Betulinic acid derivatives as HIV-1 antivirals. Trends Mol Med 2005; 11: 31-36
• 38 Oliveira-Costa JF, Meira CS, Neves MVG, Dos Reis BPZC, Soares MBP. Anti-inflammatory activities of betulinic acid: A review. Front Pharmacol 2022; 13: 883857
• 39 Gomes Castro AJ, Cazarolli LH, Bretanha LC, Sulis PM, Padilla DPR, Novoa DMA, Dambrós BF, Pizzolatti MG, Silva FRMB. The potent insulin secretagogue effect of betulinic acid is mediated by potassium and chloride channels. Arch Biochem Biophys 2018; 648: 20-26
• 40 Quan HY, Kim DY, Kim SJ, Jo HK, Kim GW, Chung SH. Betulinic acid alleviates non-alcoholic fatty liver by inhibiting SREBP1 activity via the AMPK-mTOR-SREBP signaling pathway. Biochem Pharmacol 2013; 85: 1330-1340
• 41 Li J, Jing JJ, Bai Y, Li Z, Xing R, Tan B, Ma X, Qiu WW, Du C, Du B, Yang F, Tang J, Siwko S, Liu M, Chen H, Luo J. SH479, a betulinic acid derivative, ameliorates experimental autoimmune encephalomyelitis by regulating the T helper 17/regulatory T cell balance. Mol Pharmacol 2017; 91: 464-474