Development of a New Formulation ® for Onychomycosis Treatment Using \eax Furvina® as an Active Pharmaceutical Ingredient ‘Lenin Perer Rodriguez, Yaset Rodriguer-Rodriguez, Zenaida Rodriguer-Negrin, Reinaldo Moline-Ruiz, Ricardo Medina-Marrero and Evys Ancede-Gallardo Abstract The currently available testments for onychomycosis are limited due to the absence of « formulation allowing the effective penetration of the active ‘pharmaceutical ingredient (APD. In this research, a chemical-pharmaceutical study ‘ver a new formulation for the teatment of onychomycosis, based on 2-bromo-5- (2-bromo-2-nitrovinyl furan (Furvina) composed by 0.25% of this API. 1% urea as Keratinzing agent and 9875% benzyl alcohol as pharmaceutical vehicle, was performed. This tincture proved to be active against Candida albicans. In addition, ‘an UVWVis spectrophotometric analyic technique with a maximum absorption at ‘372 nm was developed and validated. This method was specif, precise, accurate, and Linear inthe range of 2-28 ma/L. The detection and quantification limits for this technique were 06567 and 0.6636 mg/L, respectively. Keywords Onychomycosis - Furvina - Spectrophotometric method UV/Vis - Validation Highlights ‘© A new formulation for onychomycosis weatment having Furvina as an API with broad-spectrum antifungal activity has been develope. ‘© A new spectrophotometic method was developed and validated for quantifying Furvina ia the tineure. 7, Perez Rovriguez (F2) . Redrigusz Rodriguez - 7. Rosiiguee Negro. 'R. Moline Rui“ R. Medina Marrem F. Anced Gallen CCestro de Biictiv: Quinions (CBQ,, Universidad Central Marts Abreu” de Las Ville, Carers a Caran Km 5, Saota Chin CP 4890, Vila Chr. Cob femal: sereruclveda ct © Springer Nate Switzcland AG 2019 wo Cardenas etal. (els). Procedines ofthe nd Intermational Conference on BioGeostiences,itylloi on. 0.1 007/978 3-030-0428-2 17 Ree nd 2nd Intemational er) Pesta)12, 2. Perez Rodrigue «a 1 Introduction Pharmaceutical companies must insert sew formulations into the market in ordes to censure heir row, However, the cost of drug research and development (R&D) are very high and promising results are not always obtained [1 The active plar- ‘maceutical ingrediznt (APD) and its formulaion have w undergo several phases of| testing, prior to its insertion as a new drug. Consequently, the aathorzation for its ccommercalization may take over 10-15 yeas. Besides, many of these products do ‘ot amive at their final evaluation stage and have to ke abandored (2) Tinea unguiwn, onychomycosis or ngworm of the nails, isa disease caused by fungi [3]. Yeasts of the genus Candida produce damages on fingernails and wail folds witout predominance 10 any ofthe fingers. This infocion (onychomycosis is ‘unpleasant to patients due tothe incidence of this disease in social relationships and especially in the work related to food handling [4], Moreover, onychomycosis has ‘worldwide distibution and high incidence, affecting approximately 109% of the ‘general population and up to 50% of people aged over 60 years [5,6]. Many formulations to teat onychomycosis are recognized in the Cental ‘American market (2), but they have high concentrations of APIs and require lengthy treatment periods. The process of new drug development pssses through several well-defined phaces (7-9). The frst steps are the formulation studies and the chemical-pharmaceutical characterization of new drugs. Furthermore, chemical ‘methods for the determination anal quantification of APIS in the formulaions ate needed in the characterization process. Furvins” or G-l is a chemical compound named 2-bomo- (@-bromo-2-nitroviny)-furan, This substance hat a broad antimicrobial spectrum (10, 11), and it is used indifferent formulations as an antifungal and antibacterial ‘agent. Is used in the following formulations: Demofural®, Queratorurl®, and Furvinol®, which are used in human and veterinary health: and Vitrofural”, which is lized as chemical steilant in viroplnt production [12]. The aim of this ressarch is to develop a now formulation for onychomycosis treatment using Furvina® as an APL 2 Materials and Methods The Centro de Bioactivos Quimicos (CBQ) it has been proposed a new dug formulation forthe treatment of onychomycosis. This formation is aed in a tincture with Furvina® (commercial name) as API, urea, and benzyl alcohol as eratiniing agent and adjuvant, respesively.Development ofa New Formulation for Onychomycosis Treatment 193 2.1, Materials Pare samples, Furvina® standard (99.73% produced in CBQ), ethanol, rea, benz alcohol. and other reagents of analytical grade, were purchased from Merck Millipore, Germany. 2.2 Preparation of Tincture In the process of selecting a suitable solvent for making « tincture formulation, several aspects Were taken info account. It was chosen a solvent that has reported that use aS a pharmaceutical excipient [13] and must meet the requirement of being chemically inert to the other ingredienss of the formulation. As a last aspect. the excipient chosen nust have a moderate polarity that is capsble of dissolving both the active ingredicet and urea In this sclection process, benzyl alcohol was clected, ‘with a previous experimental varification that has the chemical and physical requirements to be used in our tincture formulation. ‘The tincture solution was | prepared using 0.25% — 2-bromo-5- (2-bromo-2-nitroviay! furan (Furvina®) as an APL, 1% ureaas a keatinizing agent, ‘and 98.75% benzyl alcohol as a pharmaccutical vehicle. ‘The mixture was placed in ‘an ultrasonic bath for 15 min to achieve its complete dissolution. Then, the same solvent was subsequently added to this solution in a 100 ml. volumetric fask to ‘obtain afinal concentration of Furvina of 0.25 us/mL. 2.3. Equipment ‘The main instruments used were a UV/Vis spectrophotometer with one cm matched quartz calls (model Thormo), an electronic balance (Sartaius CP 225) and an ultrasonic cleaner (BRANSON 5510). The glassware used in each procedure was Soaked overnight in a mixture of chromic acid and sulfuric acd, rinsed thoroughly ‘with double distilled water, and dried with hot air in an oven before use. The absorption spectra of reference and test solution were cartied out in quaitz cells (oe com) over the range of 190-500 nim. 24 Antifungal Activity of the Tincture ‘The antifungal activity of the tincmure was assessed by the agar diffusion method. Wells were puncture using a sterile cork borer from Sabouraud dextrose agar, previously seeded with Candida albicans ATCC 10231 as a test organism. Wells