j. res. pharm. Journal of Research in Pharmacy 2630-6344 Marmara University Pharmaceutical Delivery Technologies İlaç Dağıtım Teknolojileri Development and evaluation of azelaic acid-cyclodextrin hydrogels for treatment of acne Arpa Muhammet Davut ISTANBUL MEDIPOL UNIVERSITY, FACULTY OF PHARMACY, DEPARTMENT OF PHARMACEUTICAL TECHNOLOGY https://orcid.org/0009-0006-3223-0275 Doğan Nilay ISTANBUL MEDIPOL UNIVERSITY, FACULTY OF PHARMACY, DEPARTMENT OF PHARMACEUTICAL TECHNOLOGY https://orcid.org/0009-0001-3263-7845 Gündüz Melisa ISTANBUL MEDIPOL UNIVERSITY, FACULTY OF PHARMACY https://orcid.org/0000-0003-4968-3634 Karavelioğlu Elçin Su ISTANBUL MEDIPOL UNIVERSITY, FACULTY OF PHARMACY Erim Ümit Can ISTANBUL MEDIPOL UNIVERSITY, FACULTY OF PHARMACY, DEPARTMENT OF BASIC PHARMACEUTICAL SCIENCES, DEPARTMENT OF ANALYTICAL CHEMISTRY 06 28 2025 28 1 177 190 10 26 2023 12 11 2023 Copyright © 2010, Journal of Research in Pharmacy 2010 Journal of Research in Pharmacy

Acne is an inflammatory or non-inflammatory disease that can be seen at every stage of human life and affects daily life both physically and psychologically. In the topical treatment of acne, substances with different pharmacological effects such as benzoyl peroxide, isotretinoin, azelaic acid (AZE), and erythromycin are used alone or in combination. AZE cream (20%) has been frequently preferred in the treatment of mild and moderate acne in recent years. However, since AZE has poor solubility in water, its low permeability problems may occur. In this study, the soluble form of AZE was obtained by forming an inclusion complex of AZE with 2-hydroxypropyl β-cyclodextrin (HβC). In addition, the gel formulation, which generally exhibits higher permeability properties than oily formulations, was prepared using hydroxypropyl methylcellulose (HPMC). The formation of the inclusion complex was confirmed by NMR, XRD, DSC, and FTIR analyses. As a result of the characterization studies of the hydrogel formulations, it was determined that they had pseudoplastic flow and their viscosity was approximately 220 Pa.s. In drug release studies conducted with the dialysis bag method, it was found that the AZE-HβC hydrogel formulations had a similar cumulative release percent as HβC-free hydrogels but showed a higher cumulative release percent than the commercial cream (Azelderm, Orva, Türkiye). It was proven that the drug release of the formulations that exhibit a release profile in accordance with Higuchi kinetics was different from Azelderm according to difference (f1) and similarity (f2) factors.

 Azelaic acid acne cyclodextrin HPMC hydrogel [1] Tan AU, Schlosser BJ, Paller AS. A review of diagnosis and treatment of acne in adult female patients. Int J Womens Dermatol. 2018; 4: 56-71. https://doi.org/10.1016/j.ijwd.2017.10.006 [2] Moradi Tuchayi S, Makrantonaki E, Ganceviciene R, Dessinioti C, Feldman SR, Zouboulis CC. Acne vulgaris. Nat Rev Dis Prim. 2015; 1: 15029. https://doi.org/10.1038/nrdp.2015.29 [3] Prasad SB. Acne vulgaris: A review on pathophysiology and treatment. Asian J Pharm Clin Res. 2016; 9: 54-59. [4] Arpa MD, Seçen İM, Erim ÜC, Hoş A, Üstündağ Okur N. Azelaic acid loaded chitosan and HPMC based hydrogels for treatment of acne: Formulation, characterization, in vitro-ex vivo evaluation. Pharm Dev Technol. 2022; 27: 268-281. https://doi.org/10.1080/10837450.2022.2038620 [5] Özer S, Şenel B, Yazan Y. Preparation and in vitro evaluation of in situ gelling system containing lithium carbonate for parenteral administration. Polym Bull. 2020; 77: 599-622. https://doi.org/10.1007/s00289-019-02764-5 [6] Jung H, Kim MK, Lee JY, Choi SW, Kim J. Adhesive hydrogel patch with enhanced strength and adhesiveness to skin for transdermal drug delivery. Adv Funct Mater. 2020; 30: 1-10. https://doi.org/10.1002/adfm.202004407 [7] Üstündağ Okur N, Çağlar EŞ, Arpa MD, Karasulu HY. Preparation and evaluation of novel microemulsion-based hydrogels for dermal delivery of benzocaine. Pharm Dev Technol. 2017; 22: 500-510. https://doi.org/10.3109/10837450.2015.1131716 [8] Pourtalebi Jahromi L, Rothammer M, Fuhrmann G. Polysaccharide hydrogel platforms as suitable carriers of liposomes and extracellular vesicles for dermal applications. Adv Drug Deliv Rev. 2023; 200: 115028. https://doi.org/10.1016/j.addr.2023.115028 [9] Mahmood A, Sharif A, Muhammad F, Sarfraz RM, Abrar MA, Qaisar MN, Anwer N, Amjad MW, Zaman M. Development and in vitro evaluation of (β-cyclodextrin-g-methacrylic acid)/na+-montmorillonite nanocomposite hydrogels for controlled delivery of lovastatin. Int J Nanomedicine. 2019; 14: 5397-5413. http://doi.org/10.2147/IJN.S209662 [10] Wangsawangrung N, Choipang C, Chaiarwut S, Ekabutr P, Suwantong O, Chuysinuan P, Techasakul S, Supaphol P. Quercetin/hydroxypropyl-β-cyclodextrin ınclusion complex-loaded hydrogels for accelerated wound healing. Gels. 2022;8(9):573. https://doi.org/10.3390/gels8090573 [11] Yurtdaş G, Demirel M, Genç L. Inclusion complexes of fluconazole with β-cyclodextrin: Physicochemical characterization and in vitro evaluation of its formulation. J Incl Phenom Macrocycl Chem. 2011; 70: 429-435. https://doi.org/10.1007/s10847-010-9908-z [12] Davis ME, Brewster ME. Cyclodextrin-based pharmaceutics: Past, present and future. Nat Rev Drug Discov. 2004; 3: 1023-1035. https://doi.org/10.1038/nrd1576 [13] Bilensoy E, Rouf MA, Vural I, Şen M, Hincal AA. Mucoadhesive, thermosensitive, prolonged-release vaginal gel for clotrimazole: β-cyclodextrin complex. AAPS PharmSciTech. 2006; 7(2): E54. https:/doi.org/10.1208/pt070238 [14] Poulson BG, Alsulami QA, Sharfalddin A, El Agammy EF, Mouffouk F, Emwas A-H, Jaremko L, Jaremko M. Cyclodextrins: Structural, chemical, and physical properties, and applications. Polysaccharides. 2021; 3: 1-31. https://doi.org/10.3390/polysaccharides3010001 [15] Sherje AP, Kulkarni V, Murahari M, Nayak UY, Bhat P, Suvarna V, Dravyakar B. Inclusion complexation of etodolac with hydroxypropyl-beta-cyclodextrin and auxiliary agents: Formulation characterization and molecular modeling studies. Mol Pharm. 2017; 14: 1231-1242. https://doi.org/10.1021/acs.molpharmaceut.6b01115 [16] Pazoki-Toroudi H, Nilforoushzadeh MA, Ajami M, Jaffary F, Aboutaleb N, Nassiri-Kashani M, Firooz A. Combination of azelaic acid 5% and clindamycin 2% for the treatment of acne vulgaris. Cutan Ocul Toxicol. 2011; 30:286-291. https://doi.org/10.3109/15569527.2011.581257 [17] Webster G. Combination azelaic acid therapy for acne vulgaris. J Am Acad Dermatol. 2000; 43: 47-50. https://doi.org/10.1067/mjd.2000.108318 [18] Kainz JT, Berghammer G, Auer-Grumbach P, Lackner V, Perl-Convalexius S, Popa R, Wolfesberger B. Azelaic acid 20 % cream: effects on quality of life and disease severity in adult female acne patients. J Dtsch Dermatol Ges. 2016;14(12):1249-1259. https://doi.org/10.1111/ddg.12889 [19] Hung WH, Chen PK, Fang CW, Lin YC, Wu PC. Preparation and evaluation of azelaic acid topical microemulsion formulation: In vitro and in vivo study. Pharmaceutics. 2021; 13(3): 410. https://doi.org/10.3390/pharmaceutics13030410 [20] Kiti K, Suwantong O. The potential use of curcumin-β-cyclodextrin inclusion complex/chitosan-loaded cellulose sponges for the treatment of chronic wound. Int J Biol Macromol. 2020; 164: 3250-3258. https://doi.org/10.1016/j.ijbiomac.2020.08.190 [21] Argenziano M, Haimhoffer A, Bastiancich C, Jicsinszky L, Caldera F, Trotta F, Scutera S, Alotto D, Fumagalli M, Musso T, Castagnoli C, Cavalli R. In vitro enhanced skin permeation and retention of ımiquimod loaded in β-cyclodextrin nanosponge hydrogel. Pharmaceutics. 2019;11(3):138. https://doi.org/10.3390/pharmaceutics11030138 [22] Kaur N, Puri R, Jain SK. Drug-cyclodextrin-vesicles dual carrier approach for skin targeting of anti-acne agent. AAPS PharmSciTech. 2010; 11: 528-537. https://doi.org/10.1208/s12249-010-9411-2 [23] Ascenso A, Vultos F, Ferrinho D, Salgado A, Filho SG, Ferrari V, Simoes S, Marques HC. Effect of tretinoin inclusion in dimethyl-beta-cyclodextrins on release rate from a hydrogel formulation. J Incl Phenom Macrocycl Chem. 2012; 73: 459-465. https://doi.org/10.1007/s10847-011-0002-y [24] Manosroi J, Apriyani MG, Foe K, Manosroi A. Enhancement of the release of azelaic acid through the synthetic membranes by inclusion complex formation with hydroxypropyl-β-cyclodextrin. Int J Pharm. 2005; 293: 235-240. https://doi.org/10.1016/j.ijpharm.2005.01.009 [25] Kumar A, Rao R. Enhancing efficacy and safety of azelaic acid via encapsulation in cyclodextrin nanosponges: development, characterization and evaluation. Polym Bull. 2020; 78: 5275–5302. https://doi.org/10.1007/s00289-020-03366-2 [26] Hassan DS, Hasary HJ. The impact of viscosity on the dissolution of naproxen immediate-release tablets. J Taibah Univ Med Sci. 2023; 18: 687-695. https://doi.org/10.1016/j.jtumed.2022.12.009 [27] Permanadewi I, Kumoro AC, Wardhani DH, Aryanti N. Effect of viscosity on iron encapsulation using alginate as a carrying agent in a controlled spray drying process. Food Res. 2022; 6: 56–67. https://doi.org/10.26656/fr.2017.6(5).613 [28] Wang Z, Xiang H, Dong P, Zhang T, Lu C, Jin T, Chai KY. Pegylated azelaic acid: Synthesis, tyrosinase inhibitory activity, antibacterial activity and cytotoxic studies. J Mol Struct. 2021; 1224: 129234. https://doi.org/10.1016/j.molstruc.2020.129234 [29] Breathnach A. Azelaic acid: A new agent in the treatment of acne: history, metabolism and biochemistry. J Dermatolog Treat. 1989; 1: 7–10. https://doi.org/10.3109/09546638909094474 [30] Zimmermann F, Meux E, Mieloszynski JL, Lecuire JM, Oget N. Ruthenium catalysed oxidation without CCl4 of oleic acid, other monoenic fatty acids and alkenes. Tetrahedron Lett. 2005; 46: 3201–3203. https://doi.org/10.1016/j.tetlet.2005.03.052 [31] Cid-Samamed A, Rakmai J, Mejuto JC, Simal-Gandara J, Astray G. Cyclodextrins inclusion complex: Preparation methods, analytical techniques and food industry applications. Food Chem. 2022; 384: 132467. https://doi.org/10.1016/j.foodchem.2022.132467 [32] Cevher E, Şensoy D, Zloh M, Mülazımoğlu L. Preparation and characterisation of natamycin: γ-cyclodextrin inclusion complex and its evaluation in vaginal mucoadhesive formulations. J Pharm Sci. 2008; 97: 4319-4335. https://doi.org/10.1002/jps.21312 [33] Jordheim LP, Degobert G, Diab R, Peyrottes S, Périgaud C, Dumontet C, Fessi H. Inclusion complexes of a nucleotide analogue with hydroxypropyl-beta-cyclodextrin. J Incl Phenom Macrocycl Chem. 2009; 63: 11-16. https://doi.org/10.1007/s10847-008-9483-8 [34] Olteanu AA, Aramă C-C, Radu C, Mihăescu C, Monciu C-M. Effect of β-cyclodextrins based nanosponges on the solubility of lipophilic pharmacological active substances (repaglinide). J Incl Phenom Macrocycl Chem. 2014; 80: 17-24. https://doi.org/10.1007/s10847-014-0406-6 [35] Deuschle VCKN, Norbert Deuschle RA, Bortoluzzi MR, Athayde ML. Physical chemistry evaluation of stability, spreadability, in vitro antioxidant, and photo-protective capacities of topical formulations containing Calendula officinalis L. leaf extract. Brazilian J Pharm Sci. 2015; 51: 63-75. https://doi.org/10.1590/S1984-82502015000100007 [36] Afreen U, Fahelelbom KM, Shah SNH, Ashames A, Almas U, Khan SA, Yameen MA, Nisar N, Bin Asad MHH, Murtaza G. Formulation and evaluation of niosomes-based chlorpheniramine gel for the treatment of mild to moderate skin allergy. J Exp Nanosci. 2022; 17: 467-495. https://doi.org/10.1080/17458080.2022.2094915 [37] Dixit K, Mohapatra D, Senapati PC, Panda R, Sahu AN. Formulation development and evaluation of Lawsonia inermis extract loaded hydrogel for wound dressing application. Indian J Pharm Sci. 2022; 84. [38] Thurein SM, Lertsuphotvanit N, Phaechamud T. Physicochemical properties of β-cyclodextrin solutions and precipitates prepared from injectable vehicles. Asian J Pharm Sci. 2018; 13: 438-449. https://doi.org/10.1016/j.ajps.2018.02.002 [39] Rao GCS, Ramadevi K, Sirisha K. Effect of β-cyclodextrin on rheological properties of some viscosity modifiers. Indian J Pharm Sci. 2014; 76: 545-548. [40] Vanti G, Wang M, Bergonzi MC, Zhidong L, Bilia AR. Hydroxypropyl methylcellulose hydrogel of berberine chloride-loaded escinosomes: Dermal absorption and biocompatibility. Int J Biol Macromol. 2020; 164: 232-241. https://doi.org/10.1016/j.ijbiomac.2020.07.129 [41] Szulc-Musioł B, Siemiradzka W, Dolińska B. Formulation and evaluation of hydrogels based on sodium alginate and cellulose derivatives with quercetin for topical application. Appl Sci. 2023; 13. https://doi.org/10.3390/app13137826 [42] Jones DS, Woolfson AD, Djokic J. Texture profile analysis of bioadhesive polymeric semisolids: Mechanical characterization and investigation of interactions between formulation components. J Appl Polym Sci. 1996; 61: 2229-2234. https://doi.org/10.1002/(SICI)1097-4628(19960919)61:12<2229::AID-APP24>3.0.CO;2-0 [43] Hurler J, Engesland A, Poorahmary Kermany B, Škalko-Basnet N. Improved texture analysis for hydrogel characterization: Gel cohesiveness, adhesiveness, and hardness. J Appl Polym Sci. 2012; 125: 180-188. https://doi.org/10.1002/app.35414 [44] Arpa MD, Yoltaş A, Onay Tarlan E, Şenyüz CŞ, Sipahi H, Aydın A, Üstündağ Okur N. New therapeutic system based on hydrogels for vaginal candidiasis management: formulation–characterization and in vitro evaluation based on vaginal irritation and direct contact test. Pharm Dev Technol. 2020; 25: 1238-1248. https://doi.org/10.1080/10837450.2020.1809457 [45] Fonseca-Santos B, dos Santos AM, Rodero CF, Daflon Gremião MP, Chorilli M. Design, characterization, and biological evaluation of curcumin-loaded surfactant-based systems for topical drug delivery. Int J Nanomedicine. 2016; 11: 4553-4562. https://doi.org/10.2147/IJN.S108675 [46] Basu S, Maity S. Preparation and characterisation of mucoadhesive nasal gel of venlafaxine hydrochloride for treatment of anxiety disorders. Indian J Pharm Sci. 2012; 74: 428-433. [47] Capanema NS V, Mansur AAP, de Jesus AC, Carvalho SM, de Oliveira LC, Mansur HS. Superabsorbent crosslinked carboxymethyl cellulose-PEG hydrogels for potential wound dressing applications. Int J Biol Macromol. 2018; 106: 1218-1234. https://doi.org/10.1016/j.ijbiomac.2017.08.124 [48] Emani S, Vangala A, Buonocore F, Yarandi N, Calabrese G. Chitosan hydrogels cross-linked with trimesic acid for the delivery of 5-fluorouracil in cancer therapy. Pharmaceutics. 2023; 15(4): 1084. https://doi.org/10.3390/pharmaceutics15041084 [49] Limpongsa E, Tabboon P, Tuntiyasawasdikul S, Sripanidkulchai B, Pongjanyakul T, Jaipakdee N. Formulation and ın vitro evaluation of mucoadhesive sustained release gels of phytoestrogen diarylheptanoids from Curcuma comosa for vaginal delivery. Pharmaceutics. 2023; 15(1): 264. https://doi.org/10.3390/pharmaceutics15010264 [50] Radwan-Pragłowska J, Janus Ł, Piątkowski M, Sierakowska A, Matysek D. ZnO nanorods functionalized with chitosan hydrogels crosslinked with azelaic acid for transdermal drug delivery. Colloids Surf B Biointerfaces. 2020; 194: 111170. https://doi.org/10.1016/j.colsurfb.2020.111170 [51] Arpa MD, Ünükür MZ, Erim ÜC. Formulation, characterization and in vitro release studies of terbinafine hydrochloride loaded buccal films. J Res Pharm. 2021; 25: 667–680. https://doi.org/10.29228/jrp.58 [52] ICH. ICH Harmonised Tripartite Guideline (2005) Validation of Analytical Procedures: Text and Methodology Q2(R1). International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use, Geneva, 1-13. 2005. https://database.ich.org/sites/default/files/Q2%28R1%29%20Guideline.pdf (accessed on 24 October 2023). [53] Kumar K, Dhawan N, Sharma H, Vaidya S, Vaidya B. Bioadhesive polymers: Novel tool for drug delivery. Artif Cells, Nanomedicine Biotechnol. 2014; 42: 274-283. https://doi.org/10.3109/21691401.2013.815194 [54] Amorós-Galicia L, Nardi-Ricart A, Verdugo-González C, Arroyo-García CM, García-Montoya E, Pérez-Lozano P, Suñé-Negre JM, Suñé-Pou M. Development of a standardized method for measuring bioadhesion and mucoadhesion that ıs applicable to various pharmaceutical dosage forms. Pharmaceutics. 2022; 14(10): 1995. https://doi.org/10.3390/pharmaceutics14101995 [55] Manna S, Lakshmi US, Racharla M, Sinha P, Kanthal LK, Kumar SPN. Bioadhesive HPMC gel containing gelatin nanoparticles for intravaginal delivery of tenofovir. J Appl Pharm Sci. 2016; 6: 22-29. https://doi.org/10.7324/JAPS.2016.60804 [56] Cho CW, Choi JS, Shin SC. Enhanced local anesthetic action of mepivacaine from the bioadhesive gels. Pak J Pharm Sci. 2011; 24: 87–93. [57] Tas C, Ozkan Y, Okyar A, Savaser A. In vitro and ex vivo permeation studies of etodolac from hydrophilic gels and effect of terpenes as enhancers. Drug Deliv. 2007; 14: 453-459. https://doi.org/10.1080/10717540701603746 [58] Shamma RN, Salah Ad-din I, Abdeltawab NF. Dapsone- gel as a novel platform for acne treatment: In vitro evaluation and in vivo performance and histopathological studies in acne infected mice. J Drug Deliv Sci Technol. 2019; 54: 101238. https://doi.org/10.1016/j.jddst.2019.101238 [59] Tuǧcu-Demiröz F, Acartürk F, Erdoǧan D. Development of long-acting bioadhesive vaginal gels of oxybutynin: Formulation, in vitro and in vivo evaluations. Int J Pharm. 2013; 457: 25-39. http://dx.doi.org/10.1016/j.ijpharm.2013.09.003 [60] Arpa MD, Kesmen EE, Biltekin SN. Novel sprayable thermosensitive benzydamine hydrogels for topical application : Development , characterization , and ın vitro biological activities. AAPS Pharmscitech. 2023; 1-16. https://doi.org/10.1208/s12249-023-02674-w [61] Gollnick H, Layton A. Azelaic acid 15% gel in the treatment of rosacea. Expert Opin Pharmacother. 2008; 9: 2699–2706. https://doi.org/10.1517/14656566.9.15.2699 [62] Patel NA, Patel NJ, Patel RP. Comparative development and evaluation of topical gel and cream formulations of psoralen. Drug Discov Ther. 2009; 3: 234-242. http://www.ncbi.nlm.nih.gov/pubmed/22495634 [63] Sankar V, Praveen C, Prasanth KG, Srinivas CR, Ruckmann K. Formulation and evaluation of a proniosome hydrocortisone gel in comparison with a commercial cream. Pharmazie. 2009; 64: 731–734. https://doi.org/10.1691/ph.2009.9095 [64] Maru S, Gathu LW, Mathenge AW, Okaru AO, Kamau F, Chepkwony HK. In Vitro drug release studies of metronidazole topical formulations through cellulose membrane. East Cent African J Pharm Sci. 2012; 15: 57–62. [65] Çağlar EŞ, Karaotmarli Güven G, Üstündağ Okur N. Preparation and characterization of carbopol based hydrogels containing dexpanthenol. Ankara Univ Eczac Fak Derg. 2023; 47. https://doi.org/10.33483/jfpau.1195397 [66] Erol İ, Üstündağ Okur N, Orak D, Sipahi H, Aydın A, Özer Ö. Tazarotene-loaded in situ gels for potential management of psoriasis: biocompatibility, anti-inflammatory and analgesic effect. Pharm Dev Technol. 2020; 25: 909-918. https://doi.org/10.1080/10837450.2020.1765180 [67] Xu X, Al-Ghabeish M, Krishnaiah YSR, Rahman Z, Khan MA. Kinetics of drug release from ointments: Role of transient-boundary layer. Int J Pharm. 2015; 494: 31–39. http://doi.org/10.1016/j.ijpharm.2015.07.077 [68] de Castro DO, Tabary N, Martel B, Gandini A, Belgacem N, Bras J. Controlled release of carvacrol and curcumin: bio-based food packaging by synergism action of TEMPO-oxidized cellulose nanocrystals and cyclodextrin. Cellulose. 2018; 25: 1249-1263. https://doi.org/10.1007/s10570-017-1646-6 [69] Küçüktürkmen B, Öz UC, Bozkir A. Diklofenak sodyum yüklü polimerik nanopartiküllerin intra-artiküler uygulaması için in situ hidrojel formülasyonu. Turkish J Pharm Sci. 2017; 14: 56-64. https://doi.org/10.4274/tjps.84803 [70] Nounou MM, El-Khordagui LK, Khalafallah NA, Khalil SA. In vitro release of hydrophilic and hydrophobic drugs from liposomal dispersions and gels. Acta Pharm. 2006; 56: 311–324.