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Journal of veterinary pharmacology and therapeutics2020; 44(1); 47-57; doi: 10.1111/jvp.12903

Concentration profiles and safety of topically applied betulinic acid and NVX-207 in eight healthy horses-A randomized, blinded, placebo-controlled, crossover pilot study.

Abstract: The naturally occurring betulinic acid (BA) and its derivative NVX-207 show anticancer effects against equine malignant melanoma (EMM) cells and a potent permeation in isolated equine skin in vitro. The aim of the study was to determine the in vivo concentration profiles of BA and NVX-207 in equine skin and assess the compounds' local and systemic tolerability with the intent of developing a topical therapy against EMM. Eight horses were treated percutaneously in a crossover design with 1% BA, 1% NVX-207 or a placebo in a respective vehicle twice a day for seven consecutive days with a seven-day washout period between each formulation. Horses were treated at the neck and underneath the tail. Concentration profiles of the compounds were assessed by high-performance liquid chromatography in the cervical skin. Clinical and histopathological examinations and blood analyses were performed. Higher concentrations of NVX-207 were found in the skin compared to BA. Good systemic tolerability and only mild local adverse effects were observed in all three groups. This study substantiates the topical application of BA and NVX-207 in further clinical trials with horses suffering from EMM; however, penetration and permeation of the compounds may be altered in skin affected by tumors.
© 2020 The Authors. Journal of Veterinary Pharmacology and Therapeutics published by John Wiley & Sons Ltd.
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Publication Date: 2020-08-26 PubMed ID: 32845519DOI: 10.1111/jvp.12903Google Scholar: Lookup
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  • Journal Article
  • Randomized Controlled Trial
  • Veterinary

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research investigated the safety and effectiveness of topically applied betulinic acid (BA) and NVX-207, substances known to counter equine malignant melanoma (EMM), in healthy horses. The results indicated that application of these substances was tolerated by the horses and reached appreciable concentrations in the skin, paving the way for potential topical treatments for EMM in horses.

Explanation of the Research

The aim of this study was to test the potential of naturally occurring betulinic acid (BA) and its derivative NVX-207 to be used as a topical therapy against equine malignant melanoma (EMM). Previous studies have indicated that these compounds could have anti-cancer effects against EMM cells, and might permeate through isolated equine skin in test tube experiments.

  • The study was conducted on eight healthy horses. They received topical applications of 1% BA, 1% NVX-207 or a placebo (an inactive substance), transferred in an appropriate medium. The applications were conducted twice a day for seven consecutive days with a break of seven days between each formulation.
  • Two locations on the body, the neck and under the tail, were selected for application of the formulations. The concentration profiles of these compounds in cervical (neck) skin was then determined by high-performance liquid chromatography, an analytical technique used to separate, identify, and quantify each component in a mixture.
  • In addition to this, clinical and histopathological examinations were carried out to check for any localized or systemic adverse effects. Blood analyses were also performed to further assess systemic tolerability.

Findings of the Research

The results of the study provided important insights on the topical application of BA and NVX-207:

  • Higher concentrations of NVX-207 were found in the skin compared to BA, suggesting that NVX-207 might be more effectively absorbed by the horses’ skin.
  • The horses showed good systemic tolerability to the substances, meaning that the treatments didn’t appear to cause harmful effects to the body’s overall function.
  • Only mild local adverse effects were observed, which could indicate light irritation or reaction at the sites of application, but nothing severe or harmful.
  • While these results are promising, the researchers suggest that the absorption of the compounds could be different in skin affected by tumors. Further testing on horses with EMM will be necessary to verify the findings.

Overall, the study underlines the potential of BA and NVX-207 for use in creating a topical treatment against EMM in horses, with further investigations needed to fully understand their efficacy and safety in tumor-affected skin.

Cite This Article

APA
Weber LA, Puff C, Kalbitz J, Kietzmann M, Feige K, Bosse K, Rohn K, Cavalleri JV. (2020). Concentration profiles and safety of topically applied betulinic acid and NVX-207 in eight healthy horses-A randomized, blinded, placebo-controlled, crossover pilot study. J Vet Pharmacol Ther, 44(1), 47-57. https://doi.org/10.1111/jvp.12903

Publication

Journal of veterinary pharmacology and therapeutics
ISSN: 1365-2885
NlmUniqueID: 7910920
Country: England
Language: English
Volume: 44
Issue: 1
Pages: 47-57

Researcher Affiliations

Weber, Lisa A
  • Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany.
Puff, Christina
  • Department of Pathology, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany.
Kalbitz, Jutta
  • Biosolutions Halle GmbH, Halle (Saale), Germany.
Kietzmann, Manfred
  • Department of Pharmacology, Toxicology and Pharmacy, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany.
Feige, Karsten
  • Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany.
Bosse, Konstanze
  • Skinomics GmbH, Halle (Saale), Germany.
Rohn, Karl
  • Department of Biometry, Epidemiology and Information Processing, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany.
Cavalleri, Jessika-M V
  • University Equine Clinic, University of Veterinary Medicine Vienna, Vienna, Austria.

MeSH Terms

  • Administration, Topical
  • Animals
  • Antineoplastic Agents / administration & dosage
  • Antineoplastic Agents / adverse effects
  • Antineoplastic Agents / pharmacokinetics
  • Cross-Over Studies
  • Female
  • Horses / metabolism
  • Male
  • Pentacyclic Triterpenes / administration & dosage
  • Pentacyclic Triterpenes / adverse effects
  • Pentacyclic Triterpenes / pharmacokinetics
  • Permeability
  • Pilot Projects
  • Propanolamines / administration & dosage
  • Propanolamines / adverse effects
  • Propanolamines / pharmacokinetics
  • Triterpenes / administration & dosage
  • Triterpenes / adverse effects
  • Triterpenes / pharmacokinetics
  • Betulinic Acid

Grant Funding

  • 16KN051526 / Bundesministerium fu00fcr Wirtschaft und Energie

References

This article includes 49 references
  1. Baker JR, Leyland A. Histological survey of tumours of the horse, with particular reference to those of the skin. Veterinary Record 96(19), 419-422.
  2. Born W. Epidermal DNA synthesis in early experimental acanthosis of the guinea-pig. Archiv Für Klinische Und Experimentelle Dermatologie 236, 53-60.
    doi: 10.1007/bf00504129google scholar: lookup
  3. Bullough WS. The control of epidermal thickness. British Journal of Dermatology 87, 187-189.
    doi: 10.1111/j.1365-2133.1972.tb00307.xgoogle scholar: lookup
  4. Chang SK, Riviere JE. Effect of humidity and occlusion on the percutaneous absorption of parathion in vitro. Pharmaceutical Research 10(1), 152-155.
    doi: 10.1023/a:1018901903243google scholar: lookup
  5. Fleury C, Bérard F, Balme B, Thomas L. The study of cutaneous melanomas in Camargue-type gray-skinned horses (1): Clinical-pathological characterization. Pigment Cell Research 13(1), 39-46.
    doi: 10.1034/j.1600-0749.2000.130108.xgoogle scholar: lookup
  6. Fulda S. Betulinic acid for cancer treatment and prevention. International Journal of Molecular Sciences 9(6), 1096-1107.
    doi: 10.3390/ijms9061096google scholar: lookup
  7. Galgon T, Wohlrab W, Dräger B. Betulinic acid induces apoptosis in skin cancer cells and differentiation in normal human keratinocytes. Experimental Dermatology 14(10), 736-743.
    doi: 10.1111/j.1600-0625.2005.00352.xgoogle scholar: lookup
  8. Guy RH, Hadgraft J. Prediction of drug disposition kinetics. Journal of Pharmaceutical Sciences 73(7), 883-887.
  9. Hulsart-Billström G, Dawson JI, Hofmann S, Müller R, Stoddart MJ, Alini M, Oreffo ROC. A surprisingly poor correlation between in vitro and in vivo testing of biomaterials for bone regeneration: Results of a multicentre analysis. European Cells and Materials 31, 312-322.
    doi: 10.22203/ecm.v031a20google scholar: lookup
  10. Huyke C, Laszczyk M, Scheffler A, Ernst R, Schempp CM. Treatment of actinic keratoses with birch bark extract: A pilot study. JDDG - Journal of the German Society of Dermatology 4(2), 132-136.
    doi: 10.1111/j.1610-0387.2006.05906.xgoogle scholar: lookup
  11. Huyke C, Reuter J, Rodig M, Kersten A, Laszczyk M, Scheffler A, Schempp C. Treatment of actinic keratoses with a novel betulin-based oleogel. A prospective, randomized, comparative pilot study. Journal Der Deutschen Dermatologischen Gesellschaft 7(2), 128-133.
    doi: 10.1111/j.1610-0387.2008.06865.xgoogle scholar: lookup
  12. Johnson PJ. Dermatologic tumors (excluding sarcoids). The Veterinary Clinics of North America: Equine Practice 14(3), 625-658.
    doi: 10.1016/s0749-0739(17)30190-6google scholar: lookup
  13. Johnson W. Final report on the safety assessment of cetearyl alcohol, cetyl alcohol, isostearyl alcohol, myristyl alcohol, and behenyl alcohol. Journal of the American College of Toxicology 7(3), 395-413.
    doi: 10.1080/10915810802550835google scholar: lookup
  14. Johnson W. Final report of the amended safety assessment of glyceryl laurate, glyceryl laurate SE, glyceryl laurate/oleate, glyceryl adipate, glyceryl alginate, glyceryl arachidate, glyceryl arachidonate, glyceryl behenate, glyceryl caprate, glyceryl caprylate, glyc. International Journal of Toxicology 23(2), 55-94.
    doi: 10.1080/10915810490499064google scholar: lookup
  15. Johnson W, Bergfeld WF, Belsito DV, Hill RA, Klaassen CD, Liebler D, Andersen FA. Safety assessment of 1,2-glycols as used in cosmetics. International Journal of Toxicology 31, 147S-168S.
    doi: 10.1177/1091581812460409google scholar: lookup
  16. Kalia YN, Guy RH. Modeling transdermal drug release. Advanced Drug Delivery Reviews 48(2-3), 159-172.
    doi: 10.1016/s0169-409x(01)00113-2google scholar: lookup
  17. Liebscher G, Vanchangiri K, Mueller T, Feige K, Cavalleri JMV, Paschke R. In vitro anticancer activity of Betulinic acid and derivatives thereof on equine melanoma cell lines from grey horses and invivo safety assessment of the compound NVX-207 in two horses. Chemico-Biological Interactions 246, 20-29.
    doi: 10.1016/j.cbi.2016.01.002google scholar: lookup
  18. Lubach D, Kietzmann M. Effects of treatment with dithranol, etretinate and a combination of dithranol and etretinate on epidermal metabolism and histology. Mouse tail assay. Arzneimittel-Forschung/Drug Research 38(8), 1167-1170.
  19. Luís A, Ruela M, Perissinato AG, Esselin M, Lino DS. Evaluation of skin absorption of drugs from topical and transdermal formulations. Brazilian Journal of Pharmaceutical Sciences 52(3), 527-544.
    doi: 10.1590/s1984-82502016000300018google scholar: lookup
  20. Macgillivray KC, Sweeney RW, Piero D. Metastatic melanoma in Horses. Journal of Veterinary Internal Medicine 16, 452-456.
  21. MacKay RJ. Treatment options for melanoma of gray Horses. Veterinary Clinics of North America - Equine Practice 35(2), 311-325.
    doi: 10.1016/j.cveq.2019.04.003google scholar: lookup
  22. Mills PC, Cross SE. Regional differences in the in vitro penetration of hydrocortisone through equine skin. Journal of Veterinary Pharmacology and Therapeutics 29, 25-30.
    doi: 10.1016/j.rvsc.2006.07.015google scholar: lookup
  23. Moore JS. A therapeutic practices established in human malignant melanoma in equine malignant melanoma [Doctoral dissertation]. Virginia Polytechnic Institute and State University.
  24. Moore JS, Shaw C, Shaw E, Buechner-Maxwell V, Scarratt WK, Crisman M, Robertson J. Melanoma in horses: Current perspectives. Equine Veterinary Education 25(3), 144-151.
    doi: 10.1111/j.2042-3292.2011.00368.xgoogle scholar: lookup
  25. Nogueira SAF, Torres SMF, Malone ED, Diaz SF, Jessen C, Gilbert S. Efficacy of imiquimod 5% cream in the treatment of equine sarcoids: A pilot study. Veterinary Dermatology 17(4), 259-265.
    doi: 10.1111/j.1365-3164.2006.00526.xgoogle scholar: lookup
  26. OECD. Guidance document for the conduct of skin absorption studies. OECD Environmental Health and Safety Publications Series on Testing and Assessment No. 28 France.
    doi: 10.1787/9789264078796-engoogle scholar: lookup
  27. OECD/OCDE. Guideline for the testing of chemicals No. 428: Skin Absorption: in vitro Method (2004). France.
    doi: 10.1787/20745788google scholar: lookup
  28. Patterson-Kane JC, Sanchez LC, Uhl EW, Edens LM. Disseminated metastatic intramedullary melanoma in an aged grey horse. Journal of Comparative Pathology 125(2-3), 204-207.
    doi: 10.1053/jcpa.2001.0481google scholar: lookup
  29. Phillips JC, Lembcke LM. Equine melanocytic tumors. Veterinary Clinics of North America - Equine Practice 29(3), 673-687.
    doi: 10.1016/j.cveq.2013.08.008google scholar: lookup
  30. Prausnitz MR, Elias PM, Franz TJ, Schmuth M, Tsai J-C, Menon GK, Feingold KR. Skin barrier and transdermal drug delivery. Medical Therapy 5(21), 2065-2073.
  31. Reed S, Bayly WM, Sellon D. Equine internal medicine (4th ed.). St. Louis, MO: Elsevier Inc..
  32. Rodríguez F, Forga J, Herráez P, Andrada M, Fernández A. Metastatic melanoma causing spinal cord compression in a horse. Veterinary Record 142(10), 248-249.
    doi: 10.1136/vr.142.10.248google scholar: lookup
  33. Scott DW, Miller WH. Equine dermatology (2nd ed.). Maryland Heights, MO: Elsevier Saunders.
  34. Seltenhammer MH, Simhofer H, Scherzer S, Zechner P, Curik I, Sölkner J, Eisenmengr E. Equine melanoma in a population of 296 grey Lipizzaner horses. Equine Veterinary Journal 35(2), 153-157.
    doi: 10.2746/042516403776114234google scholar: lookup
  35. 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. Journal of Investigative Dermatology 114(5), 935-940.
    doi: 10.1046/j.1523-1747.2000.00972.xgoogle scholar: lookup
  36. Smith SH, Goldschmidt MH, McManus PM. A comparative review of melanocytic neoplasms. Veterinary Pathology 39(6), 651-678.
  37. Soe L, Wurz GT, Mäenpää JU, Hubbard GB, Cadman TB, Wiebe VJ, DeGregorio MW. Tissue distribution of transdermal toremifene. Cancer Chemotherapy and Pharmacology 39(6), 513-520.
    doi: 10.1007/s002800050607google scholar: lookup
  38. Sutton RH, Coleman GT. Melanoma and the greying horse. Rural Industries Research and Development Corporation 97(55), 1-27.
  39. Traub-Dargatz JL, Salman MD, Voss JL. Medical problems of adult horses, as ranked by equine practitioners. Journal of the American Veterinary Medical Association 198(10), 1745-1747.
  40. Valentine BA. Equine melanocytic tumors: A retrospective study of 53 Horses (1988 to 1991). Journal of Veterinary Internal Medicine 9(5), 291-297.
    doi: 10.1111/j.1939-1676.1995.tb01087.xgoogle scholar: lookup
  41. Valentine B. Survey of equine cutaneous neoplasia in the Pacific Northwest. Journal of Veterinary Diagnostic Investigation 18, 123-126.
  42. VICH. Target animal safety for veterinary pharmaceutical products (VICH GL 43). European Medicines Agency - Veterinary Medicines and Inspections .
    doi: 10.1007/978-3-0346-0295-2_2google scholar: lookup
  43. Weber LA, Funtan A, Paschke R, Delarocque J, Kalbitz J, Meißner J, Cavalleri J-M-V. In vitro assessment of triterpenoids NVX-207 and betulinyl-bis-sulfamate as a topical treatment for equine skin cancer. PLOS ONE under review.
  44. Weber LA, Meißner J, Delarocque J, Kalbitz J, Feige K, Kietzmann M, Cavalleri J-M-V. Betulinic acid shows anticancer activity against equine melanoma cells and permeates isolated equine skin in vitro. BMC Veterinary Research 16(44), 1-9.
    doi: 10.1186/s12917-020-2262-5google scholar: lookup
  45. Willmann M, Wacheck V, Buckley J, Nagy K, Thalhammer J, Paschke R, Selzer E. Characterization of NVX-207, a novel betulinic acid-derived anti-cancer compound. European Journal of Clinical Investigation 39(5), 384-394.
    doi: 10.1111/j.1365-2362.2009.02105.xgoogle scholar: lookup
  46. Wilson DA, Adelstein EH, Keegan KG, Barrett BA, Kutz RR. In vitro and in vivo effects of activated macrophage supernatant on distal limb wounds of ponies. American Journal of Veterinary Research 57(8), 1220-1224.
  47. Wong D, Buechner-Maxwell V, Manning T. Equine skin: structure, immunologic function, and methods of diagnosing disease. Compendium : Continuing Education for Veterinarians-North American Edition 27(6), 463-473.
  48. Yao M, Gu C, Doyle FJ, Zhu H, Redmond RW, Kochevar IE. Why is rose Bengal more phototoxic to fibroblasts in vitro than in vivo?. Photochemistry and Photobiology 90(2), 297-305.
    doi: 10.1111/php.12215google scholar: lookup
  49. Zalesińska MD, Borska S. Betulin and its derivatives - precursors of new drugs. World Scientific News 127(3), 123-138.

Citations

This article has been cited 5 times.
  1. Pimenta J, Prada J, Cotovio M. Equine Melanocytic Tumors: A Narrative Review. Animals (Basel) 2023 Jan 10;13(2).
    doi: 10.3390/ani13020247pubmed: 36670786google scholar: lookup
  2. Yi Z, Gao Y, Yu F, Zhu Y, Liu H, Li J, Murua Escobar H. Interventions for treatment of cutaneous melanoma in horses: a structured literature review. Vet Res Commun 2023 Jun;47(2):347-360.
    doi: 10.1007/s11259-022-10023-8pubmed: 36329228google scholar: lookup
  3. Weber LA, Delarocque J, Feige K, Kietzmann M, Kalbitz J, Meißner J, Paschke R, Cavalleri JV. Effects of Topically Applied Betulinic Acid and NVX-207 on Melanocytic Tumors in 18 Horses. Animals (Basel) 2021 Nov 13;11(11).
    doi: 10.3390/ani11113250pubmed: 34827981google scholar: lookup
  4. Weber LA, Funtan A, Paschke R, Delarocque J, Kalbitz J, Meißner J, Feige K, Kietzmann M, Cavalleri JV. In vitro assessment of triterpenoids NVX-207 and betulinyl-bis-sulfamate as a topical treatment for equine skin cancer. PLoS One 2020;15(11):e0241448.
    doi: 10.1371/journal.pone.0241448pubmed: 33151949google scholar: lookup
  5. Zscherpe P, Kalbitz J, Weber LA, Paschke R, Mäder K, von Rechenberg B, Cavalleri JV, Meißner J, Klein K. Potent drug delivery enhancement of betulinic acid and NVX-207 into equine skin in vitro - a comparison between a novel oxygen flow-assisted transdermal application device and microemulsion gels. BMC Vet Res 2024 May 16;20(1):202.
    doi: 10.1186/s12917-024-04064-1pubmed: 38755639google scholar: lookup
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