Abstract
Fungal infections are a growing global health problem. Therefore, our group has synthetized and characterized an improved antimycotic by co-crystallization of ketoconazole and para-amino benzoic acid, named KET-PABA. The aim was to increase bioavailability, biocompatibility, and efficiency of the parent drug–ketoconazole. Based on our previous results showing the cocrystal improved physical properties, such as stability in suspension, solubility, as well as antimycotic efficiency compared to ketoconazole, the current study investigated the local possible side effects induced on the skin of BALBc mice by the application of KET-PABA cocrystal, in view of a further use as a topically applied antimycotic drug. A specific test (mouse ear-swelling test) was used, combined with the histopathological examination and the measurement of pro and anti-inflammatory cytokines and inflammation mediators. KET-PABA application was safe, without signs of skin sensitization shown by the mouse ear sensitization test, or histopathology. KET-PABA strongly inhibited proinflammatory cytokines such as IL1 α, IL1 β, IL6 and TNF α, and other proinflammatory inducers such as NRF2, compared to vehicle. KET-PABA had no effect on the levels of the anti-inflammatory cytokine IL10, or proinflammatory enzyme COX2 and had minimal effects on the activation of the NF-κB pathway. Overall, KET-PABA application induced no sensitization, moreover, it decreased the skin levels of proinflammatory molecules. The lack of skin sensitization effects on BALBc mice skin along with the inhibition of the proinflammatory markers show a good safety profile for topical applications of KET-PABA and show promise for a further clinical use in the treatment of cutaneous mycosis.
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References
Agilent (2010) CrysAlis PRO Agilent Technologies. Oxfordshire, England https://www.agilent.com/cs/library/usermanuals/Public/CrysAlis_Pro_User_Manual.pdf. Accessed 25 January 2021.
Ali ZE, Gerbeix C, Hemon P, Esser PR, Martin SF, Pallardy M, Kerdine-Römer S (2013) Allergic skin inflammation induced by chemical sensitizers is controlled by the transcription factor Nrf2. Toxicol Sci 134:39–48
Aronson JK (2016) Meyler’s side effects of drugs. In: Aronson JK (ed) Sunscreens, 16th edn. Elsevier, Waltham, MA, pp 603–604
Choi FD, Juhasz MLW, Mesinkovska NA (2019) Topical ketoconazole: a systematic review of current dermatological applications and future developments. J Dermatolog Treat 30:760–771
Crisan ME, Bourosh P, Maffei ME, Forni A, Pieraccini S, Sironi M, Chumakov YM (2014) Synthesis, crystal structure and biological activity of 2-hydroxyethylammonium salt of p-aminobenzoic acid. PLoS ONE 9:e101892. https://doi.org/10.1371/journal.pone.0101892
David L, Moldovan B, Vulcu A et al (2014) Green synthesis, characterization and anti-inflammatory activity of silver nanoparticles using European black elderberry fruits extract. Colloids Surf B Biointerfaces 122:767–777
Deng P, Teng F, Zhou F, Song Z, Meng N, Liu N, Feng R (2017) Y-shaped methoxy poly (ethylene glycol)-block-poly (epsilon-caprolactone)-based micelles for skin delivery of ketoconazole: in vitro study and in vivo evaluation. Mater Sci Eng C Mater Biol Appl 78:296–304
Dolomanov OV, Bourhis LJ, Gildea RJ, Howard JAK, Puschmann H (2009) OLEX2: a complete structure solution, refinement and analysis program. J Appl Crystallogr 42:339–341
Dunn BJ, Rusch GM, Siglin JC, Blaszcak DL (1990) Variability of a mouse ear swelling test (MEST) in predicting weak and moderate contact sensitization. Fundam Appl Toxicol 15:242–248. https://doi.org/10.1016/0272-0590(90)90051-k
Emter R, Ellis G, Natsch A (2010) Performance of a novel keratinocyte-based reporter cell line to screen skin sensitizers in vitro. Toxicol Appl Pharmacol 245:281–290
Gad SC, Brendan J, Dunn BJ, Dobbs DW, Reilly C, Walsh RD (1986) Development and validation of an alternative dermal sensitization test: The mouse ear swelling test (MEST). Toxicol and Applied Pharmacol 84:93–114. https://doi.org/10.1016/0041-008X(86)90419-9
Garrigue JL, Nicolas JF, Fraginals R, Benezra C, Bour H, Schmitt D (1994) Optimization of the mouse ear swelling test for in vivo and in vitro studies of weak contact sensitizers. Contact Dermatitis 30:231–237
Glaser DA, Prodanovic E (2016) Sunscreens. In: Draelos ZD, Dover JS, Alam M (eds) Cosmeceuticals, 3rd edn. Elsevier, Philadelphia, PA, pp 143–153
Greenspoon J, Ahluwalia R, Juma N, Rosen CF (2013) Allergic and photoallergic contact dermatitis: a 10-year experience. Dermatitis 24:29–32
Helou DG, Martin SF, Pallardy M, Chollet-Martin S, Kerdine-Röme S (2019) Nrf2 involvement in chemical-induced skin innate immunity. Front Immunol 10:1004
Ho AW, Thomas S. Kupper (2019) Soluble mediators of the cutaneous immune system. In: Kang S, Amagai M, Bruckner AL, Enk AH, Margolis DJ, McMichael AJ, Orringer JS (eds) Fitzpatrick’s Dermatology, ninth edition. McGraw-Hill Education, pp 165, 174
Hu ML, Chen YK, Chen LC, Sano M (1995) Para-aminobenzoic acid scavenges reactive oxygen species and protects DNA against UV and free radical damage. J Nutr Biochem 6:504–508
Husain M, Boermans HJ, Karrow NA (2011) Mesenteric lymph node transcriptome profiles in BALB/c mice sensitized to three common food allergens. BMC Genom 6(12):12. https://doi.org/10.1186/1471-2164-12-12
Josefowicz SZ, Lu LF, Rudensky AY (2012) Regulatory T cells: mechanisms of differentiation and function. Annu Rev Immunol 30:531–564
Kadhum WR, Oshizaka T, Ichiro H, Todo H, Sugibayashi K (2016) Usefulness of liquid–crystal oral formulations to enhance the bioavailability and skin tissue targeting of p-amino benzoic acid as a model compound. Eur J Pharm Sci 88:282–290
Khurana A, Sardana K, Chowdhary A (2019) Antifungal resistance in dermatophytes: recent trends and therapeutic implications. Fungal Genet Biol 132:103255. https://doi.org/10.1016/j.fgb.2019.103255
Kissenpfennig A, Henri S, Dubois B, Laplace-Builhé C, Perrin P, Romani N, Tripp CH, Douillard P, Leserman L, Kaiserlian D, Saeland S, Davoust J, Malissen B (2005) Dynamics and function of Langerhans cells in vivo: dermal dendritic cells colonize lymph node areas distinct from slower migrating Langerhans cells. Immunity 22:643–654
Kluzk A, Popek T, Kiyota T, Macedo P, Stefanowicz P, Lazar C, Konishi Y (2002) Drug evolution: p-aminobenzoic acid as a building block. Curr Med Chem 9:1871–1892
LaBerge L, Pratt M, Fong B, Gavigan G (2011) A 10-year review of p-phenylenediamine allergy and related para-amino compounds at the Ottawa patch test clinic. Dermatitis 22:332–334
Lauritanoa D, Ronconib G, Caraffac A, Gallengad CE, Kritase SK, Di Emidiof P, Martinottig S, Tetèh G, Rossi R, Conti P (2020) New aspect of allergic contact dermatitis, an inflammatory skin disorder mediated by mast cells: can IL-38 help? Med Hypotheses 139:109687
Li W, Wu X, Xu X, Wang W, Song S, Liang K, Yang M, Guo L, Zhao Y, Li R (2016) Coenzyme Q10 suppresses TNF- α-induced inflammatory reaction in vitro and attenuates severity of dermatitis in mice. Inflammation 39:281–289. https://doi.org/10.1007/s10753-015-0248-4
Mackie BS, Mackie LE (1999) The PABA story. Australas J Dermatol 40:51–53
Martin F, Pop M, Kacso I, Grosu IG, Miclăuș M, Vodnar D, Lung I, Filip GA, Olteanu ED, Moldovan R, Nagy A, Filip X, Bâldea I (2020) Ketoconazole-p-aminobenzoic acid cocrystal: revival of an old drug by crystal engineering. Mol Pharm 17:918–932
Noble JE, Bailey MJ (2009) Quantitation of protein. Methods Enzymol 463:73–95. https://doi.org/10.1016/S0076-6879(09)63008-1
Ricciotti E, Garret AGA (2011) Prostaglandins and inflammation. Arterioscler Thromb Vasc Biol 31:986–1000
Sebastiani S, Albanesi C, De Pità O, Puddu P, Cavani A, Girolomoni G (2002) The role of chemokines in allergic contact dermatitis. Arch Dermatol Res 293:552–559
Sharma RS, Joy RC, Boushey CJ, Ferruzzi MG, Leonov AP, McCrory MA (2013) Effects of para-aminobenzoic acid (PABA) form and administration mode on PABA recovery in 24-hour urine collections. J Acad Nutr Diet 114:457–463
Sheldrick GM (1997) SHELXS97 and SHELXL97. University of Göttingen, Germany
Sheldrick GM (2008) A short history of SHELX. Acta Crystallogr SEct A: Found Crystallogr 64:112–122
Stewart I, Seawright AA, Schluter PJ, Shaw GR (2006) Primary irritant and delayed-contact hypersensitivity reactions to the freshwater cyanobacterium Cylindrospermopsis raciborskii and its associated toxin cylindrospermopsin. BMC Dermatol 6:5. https://doi.org/10.1186/1471-5945-6-5
Biovia Materials Studio, release 5.5., BIOVIA (previously named Accelrys), USA https://www.3ds.com/products-services/biovia/products/molecular-modeling-simulation/biovia-materials-studio.
Suzuki K, Meguro K, Nakagomi D, Nakajima H (2017) Roles of alternatively activated M2 macrophages in allergic contact dermatitis. Allergol Int 66:392–397
Tak PP, Firestein GS (2001) NF-κB: a key role in inflammatory diseases. J Clin Invest 107:7–11
Thorne PS, Hawk C, Kaliszewski SD, Guiney PD (1991) The non-invasive mouse ear swelling assay: I. Refinements for detecting weak contact sensitizers. Fundam APpl ToxIcol 17:807–820
Thune P (1984) Contact and photocontact allergy to sunscreens. Photodermatol 1:5–9
Tordesillas L, Goswami R, Benedé S, Grishina G, Dunkin D, Järvinen KM, Maleki SJ, Sampson HA, Berin MC (2014) Skin exposure promotes a Th2-dependent sensitization to peanut allergens. J Clin Invest 124:4965–4975
Wang N, Liang H, Zen K (2014) Molecular mechanisms that influence the macrophage m1–m2 polarization balance. Front Immunol 5:614
Zhang M, Zhou J, Wang L, Li B, Guo J, Guan X, Han Q, Zhang H (2014) Caffeic acid reduces cutaneous tumor necrosis factor alpha (TNF- α), IL-6 and IL-1 β levels and ameliorates skin edema in acute and chronic model of cutaneous inflammation in mice. Biol Pharm Bull 37:347–354
Funding
This work was supported by a grant of the Romanian National Authority for Scientific Research and Innovation, CNCS/CCCDI–UEFISCDI, project number PN–III-P2-2.1-PED-2016–1521 (157PED), within PNCDI III.
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Conceptualization of the research was made by S.D. and I.B. I.B. and R.M. carried out the experiments and contributed to the sample preparation. D.O. and I.B. contributed to the western blot analysis. F.A. performed the statistical analysis. X.F., F.M., I.K. were responsible for the preparation and characterization of the compound tested. A.N. performed the histopathological analysis. All authors approved the final article. The manuscript was written through the contributions of all authors. All authors have given approval to the final version of the manuscript.
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All lab animal experiments were approved by ethics committee of the University of Medicine and Pharmacy Cluj-Napoca and the Veterinary Health Directorate, Romania (authorization number 67/ 06 06 2017).
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Danescu, S., Filip, G.A., Moldovan, R. et al. Ketoconazole-p aminobenzoic cocrystal, an improved antimycotic drug formulation, does not induce skin sensitization on the skin of BALBc mice. Inflammopharmacol 29, 721–733 (2021). https://doi.org/10.1007/s10787-021-00834-7
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DOI: https://doi.org/10.1007/s10787-021-00834-7