Oral Fast Disintegrating Anti-allergic Film Using Hot-melt

Oral Fast Disintegrating Anti- solelyergic Film exploitation Hot-melt Development and Evaluation of an Oral Fast Disintegrating Anti-allergic Film Using Hot-melt Extrusion TechnologyManjeet B. Pimparade1, Anh Vo1, Abhijeet S. Maurya1, Joseph T. Morott1, Xin Feng1, Dong Wuk Kim1, Vijay I. Kulkarni1, Roshan Tiwari1, K. Vanaja2, Reena. Murthy2, H. N. Shivakumar2, D. Neupane3, S. R. Mishra3, S. N. Murthy1, Michael. A. Repka1,4*1Department of Pharmaceutics medicine Delivery, School of Pharmacy, The University of Mississippi, University, MS, USA2Institute for Drug Delivery and Biomedical Research, Bangalore, India3Department of Physics, University of Memphis, Memphis, TN, USA4Pii Center for Pharmaceutical Technology, The University of Mississippi, University, MS, USA* forebode for correspondenceMichael A. Repka, D.D.S., Ph.D.Professor and Chair, Department of Pharmaceutics and Drug DeliveryDirector, Pii Center for Pharmaceutical TechnologySchool of PharmacyThe University of Mississippi , University, MS 38677Abstract The main target of this novel culture was to develop chlorpheniramoe maleate viva vocely disintegrating films (ODF) using hot-melt jutting technology and evaluate the characteristics of the preparation using in vitro and in vivo methods. limited starch with glycerine was used as a polymer matrix for melt jut. Sweetening and saliva-simulating agents were incorporated to improve palatability and lower the disintegration age of film formulations. A ensample screw configuration was applied, and the last geographical zone of the barrel was opened to discharge wet vapors, which helped to manufacture non-sticky, clear, and uniform films. The film formulations demonstrated speedy disintegration clock (6-11 s) and more than 95% dis result in 5 second. In addition, the films had characteristic automatonlike properties that were helpful in handling and warehousing. An animal model was employed to do the test masking of melt- produced films. The le ad film formulation was subjected to a human panel for evaluation of extent of relish masking and disintegration.Keyword Hot-melt extrusion, Orally disintegrating film, Taste-masking, Pediatric and geriatric, Modified starch, Human panel understanding evaluation, Continuous manufacturing, Chlorpheniramine maleateIntroductionIt is estimated that 26-50% of the patient population find difficulty in swallowe tablets and hard gelatin capsules 1. These patients mainly include the decrepit who eat difficulty taking conventional oral social disease forms because of hand tremors and dysphagia, and pediatric patients who are often fearful of taking solid oral loony toons forms owing to their underdeveloped muscular and nervous systems 2. In addition, patients who are mentally ill, developmentally disabled, uncooperative, on reduced liquid-intake plans or nauseated, and travelers who may not have coming to clean wet system also are substructuredidates for ODFs 3, 4.The traditional alt ernative to swallowing difficulties is formulating a drug substance in liquid dosage form. However, liquid dosage forms have several limitations, such as the need for measuring, bulkiness, physical, chemical, and microbial stability issues, spoilage, inaccurate dosing, and organoleptic properties of drug and drug formulations 5.Conventional solid oral formulations contri exactlyed significantly to minimizing the shortcomings of liquid dosage forms. The crushing of tablets or opening of capsules is a straightforward way for patients or caregivers to lessen the swallowing difficulties. However, serious consequences may be associated with special- vacate, enteric-coated, and cytotoxic or hormonal medicines, as these formulations are designed for special cases 6. Moreover, European Medical Agency does not recommend the splitting or crushing of tablets because the active pharmaceutical ingredient (API) is not evenly distributed in the tablet 7, 8. Thus, it is very convenient to develop a formulation that disintegrates in the oral cavity and eases the swallowing process.In recent years, fast disintegrating oral formulations established their importance in patient population suffering from dysphagia, stroke, thyroid disorder, Parkinsons disease, multiple sclerosis, and cerebral palsy 9. Commercially available orodispersible tablets (ODT) and orodispersible films (ODF) are the most palmy platforms for pharmaceutical product development. ODTs are solid oral dosage forms that disintegrate rapidly, typically within 30 s, with or without the governance of additional pee 10. They provided abundant comfort to patients with swallowing difficulties 11. Despite the benefits of ODTs, there are some challenges in their processing and handling owing to their fragility and brittleness, which warrant special package for protection during storage and transportation 12. The films are flexible and not as fragile as most ODTs. Hence, there is ease in transportation, consumer handl ing, and storage of ODFs.ODF can be define as a dosage form that employs a water system-soluble polymer (generally a hydrocolloid, which may be a bioadhesive polymer), which allows the dosage form to quickly wet, adhere, and dissolve to release the drug when set(p) on the expression or in the oral cavity 5. ODF alleviated patient discomforts associated with swallowing disabilities without compromising the therapeutic effect. In addition, it could ease the administration of drugs to pediatric patient population 13. Moreover, ODF can be helpful in curtailing dose variations in younger patients, in whom liquid formulations are the most original way of drug delivery.Currently, solvent casting methods are commonly employed to produce ODFs, owing to its ease of production and low set up cost 14, 15. Despite its wide application, products with batch-to-batch variation may be produced because of multiple steps involved in the production. In addition, air entrapment in the films is common ly observed in solvent casting methods, which leads to dose variations and inappropriate esthetic appearance of the product 15. The use of large numerates of solvent is one of the biggest shortcomings of this method as solvent removal and disposal is a long and tedious process. Thus, it is very beneficial to develop a solvent-free, quick, and continuous process that could diminish the shortcomings of the current manufacturing method.Hot melt extrusion (HME) is a one-step, solvent-free continuous manufacturing process, which established itself in the pharmaceutical arena for the development of various solid oral formulations 16-25. This technology involves the use of temperature and shear to process polymer blends and extrude them through a die of the desired design 26. HME could be an effective alternative to the solvent casting method as it diminishes the inherent shortcomings, such as the use of solvents and problems involved in the mixing and drying steps. This ultimately makes HME process efficient and cost effective for patients 27, 28.This study has three main objectives to 1) develop a chesty patient-friendly orally fast disintegrating film of chlorpheniramine maleate (CPM) 2) evaluate these formulations with different in vitro and in vivo techniques, and 3) demonstrate the feasibility of HME techniques for continuous manufacturing of ODF without the use of solvents. To the best of our knowledge, there is no promulgated literature on the manufacturing of orally fast disintegrating formulations using HME technology and evaluation of films using in vitro and in vivo techniques.Materials and Methods 2.1. MaterialsCPM was purchased from MP Biomedicals, LLC (Solon, OH, USA). Lycoat RS 780 (modified starch) was supplied by Roquette America Inc. (Keokuk, IA, USA). citric acid and glycerol were ordered from Fisher Scientific (Pittsburgh PA, USA). Magnasweet sample was gifted by Mafco worldwide LLC (Camden, NJ, USA). Sucralose was supplied by JK Sucralose Inc . (Edison, New Jersey, USA).2.2. Thermal abstractThermogravimetric depth psychology (TGA) studies (Perkin Elmer Pyris 1, Shelton, CT, USA) were performed to estimate the thermal stability of the API and excipients during HME processing. Data were analyzed using Pyris software. The API excipients were heated from 30-160 at 20 /min.2.3. Material preparation and blending CPM, citric acid, and Lycoat RS 780 were dry miscellaneous at amounts outlined in Table 1 using a V-shell blender (GlobePharma, Maxiblend, New Brunswick, NJ, USA) after passing through an ASTM 30 mesh. The plasticizer (glycerol with fade away sucralose and Magnasweet) was incorporated slowly into a risque-shear mixer (Model RSI 3VG, Robot Coupe Industrial Division, Ridgeland, MS, USA) containing the previously mixed blend with all excipients and allowed to blend for 10 min.2.4. Hot melt extrusion The blends were melt-extruded using a co-rotating twin-screw extruder (16 mm Prism EuroLab, ThermoFisher Scientific, Pit tsburgh, PA, USA) at 30-50 rpm over a temperature range of 100- 110. A degassing port was deliverd in the last zone of the barrel to release excess water vapor, which would otherwise produce unwanted bubbles in the films. Additionally, the film die was installed with preset thickness. The physical blend of the formulation was manually fed into the hopper, and the films were collected, wrapped in wax paper, sealed, and stored in polyethylene bags at 25 with 40% congress humidity.2.5. Film characterizations2.5.1 Film thickness and mechanical propertiesThe mechanical properties of the films were evaluated using the TA.XTPlus texture analyzer equipped with 5 kg load cell (Texture Technologies, Scarsdale, NY, USA). The films were cut into dumbbell shaped specimens with a width and length of 1.55 and 15.5 mm, respectively, and placed longitudinally in tensile grip probe on the texture analyzer. The films were tested at a crosshead move of 2 mm/min and held between two clamps positioned at 5 mm. The results of film samples that broke at and not between the clamps were not included in the calculations. Each film formulation was measured with ten replicates 29. The tensile strength (Ts) and percent elongation (%E) were reckon using the results from texture analyzer. Film thickness was measured using an electronic calliper (Fisher Scientific, Pittsburgh, PA, USA) at different positions.2.5.2 Disintegration test The film was cut into an appropriate size as per the dose (4 mg) and placed in a petri dish. Then, 100 L artificial salivary media was added, and the time for complete disintegration of the film was recorded (n =10).2.5.3 Surface pH of film The film was moistened using 5 L water and a contact electrode touched the show up of the film (Oakton pH meter, Fisher Scientific, Pittsburgh, PA, USA), followed by pH measurement (n=6)2.6. Analytical methodA Waters game performance liquid chromatography (HPLC) system equipped with a Water 600 binary pump, Waters 2489 UV/detector, and Waters 717 plus autosampler (Waters Technologies Corporation, Milford, MA, USA), and a Phenomenex Luna 5 m C18 (2) 250 x 4.6 mm column (Torrance, CA, USA) were used at a detection wavelength of 254 nm. The mobile phase consisted of 7.5 mM monobasic potassium phosphate in methanol and water at a ratio of 62.537.5 (v/v). The mobile phase stop rate was maintained at 1.0 mL/min, and an injection volume of 10 L was used 30. HPLC entropy were analyzed using Empower 2 software (Milford, MA, USA).2.7. In vitro dissolution studiesThe films for dissolution studies were cut into sizes relative to the dose of CPM (4 mg). The drug profile was evaluated using a USP dissolution apparatus-I (Hanson SR8, Chatsworth, CA) maintained at 37 0.5 and having a shaft rotation speed of 100 rpm. The dissolution test was performed using 900 mL phosphate weaken (pH 6.8). The samples were withdrawn at 5, 10, and 30 min and analyzed using the HPLC- UV system.2.8. roentgen ray diffraction stu dies (XRD)X-Ray diffraction (Bruker D8 Advance, Madison, MI, USA) was used to determine the physical state of the drug, excipients, and film formulations. The X-ray diffraction apparatus used CuK radiation at 40 mA, 40 kV, a scanning speed of 2-/min, and diffraction angle (2) range of 5-55.2.9. Scanning electron Microscope (SEM)The surface morphology of the films was evaluated using SEM analysis. The samples were mounted on adhesive carbon pads placed on aluminum and sputter coated with gold using a heater sputtering system (Anatech Ltd, Springfield, VA, USA) in a high vacuum evaporator. A JEOL JSM-5600 SEM operating at an accelerating voltage of 10 kV was used for imaging.2.10. In vivo degustation evaluation Twenty-one nave adult male Sprague-Dawley rats (175-200 g) were ordered from Harlan Laboratories (Houston, TX, USA) for the study. The rats were housed in Plexiglass cages with Corncob bedding in a vivarium that maintained a 12 h light/dark cycle and an ambient temperature of 22. Food and water were available without any restriction, except during the training and taste evaluation experiments as mentioned below (2.10.1). All surgical procedures were approved by the Institutional Animal Care and Use Committee (IACUC) at The University of Mississippi, University, USA (protocol no. 15-026). This study was performed as per the procedure in our previous publication on taste assessment method for bitter drugs 31.2.10.1. Training paradigm The rats were trained for licking behavior (response to thirst) by depriving them of water for 22 h, but they had ad libitum overture to food. by and by the water deprivation period, the Plexiglass cage was divided using plastic transparent dividers to provide an person water bottle to each animal. Eventually, the rats were provided with graduated water bottle for 30 min, and the amount consumed at 15 and 30 min were recorded. This training paradigm was performed for 2 days before the taste evaluation experiment.2.10.2. E valuation of bitterness sensitivity of ratsTo determine the concentration of CPM for this study, a sensitivity test for bitterness was performed in rats. After depriving the rats of water for 22 h, sensitivity toward 0.5 mg/mL CPM solution was evaluated on the first day, followed by a washout period of 24 h. Subsequently, the effect of 1 mg/mL CPM solution was examined, and the results were recorded.2.10.3. Experiment The experiment was performed for 30 min with 30 mL test formulation following the 22 h water deprivation period. After each experiment, the rats had a washout period of 24 h to avoid any memory of the taste of the previous formulation. The rats had ad libitum access to food during the experiment and washout period. The amount of solution remaining at 15 and 30 min was noted and subtracted from the original test volume. Varying results caused by spilling of the test solution while measuring or leaking of bottle knob were omitted from the study. Notably, animal behavior responses such as jaw smacking, oral grooming, and retreating were observed, which was not the sharpen of this study. Formulations N2, N7, and N9 at 0.5 mg/mL CPM were used for bitterness evaluation study in rat model. The average amount of solution consumed by all animals was calculated and expressed as the mean standard deviation. The mean scores between the physical mixture and formulation were compared using a student t-test at 95% confidence level and P2.11. Film evaluation by human panelThe evaluation of film for palatability, disintegration time (DT), and organoleptic characteristics was performed at the Institute for Drug Delivery and Biomedical Research, Bangalore India (Protocol number VIPS/2013/12). The subjects were recruited after obtaining informed consent. This study is also in accordance with the Code of Ethics of the World Medical Association (Declaration of Helsinki). The data-based procedure for this study was as per our previously published work 10, 32. 2.11.1. Human subject selection criteria Six human subjects belonging to either sex were recruited. They were asked to forbear from coffee/tea and other beverages for 12 h. The subjects were allowed to drink only water for 12 h. Moreover, they were asked not to eat chocolates or other candies for over 6 h. The cellular inclusion criterion was healthy human subjects aged 18-42 years, and the exclusion criteria were subjects suffering from fever, mouth ulcers, dry mouth, cold, nose block, and wounds as well as smokers.2.11.2. Data collection ahead data collection, the subjects were asked to wash their mouth with water at ambient temperature. The surface temperature of the tongue was recorded using an infrared (IR) thermometer, and a difference of 5 relative to the body temperature was considered an exclusion criteria.2.11.2.1. Bitterness perceptionThe subjects were asked to taste aqueous solutions of CPM, beginning with very dilute solutions and progressing to higher concentrations, by plac ing 2 mL solution for 30 s on the tongue/buccal cavity. The concentrations screened were 0, 0.5, 1, 2.5, and 4 mg. The volunteers were asked to report the perception each time 1- I feel bitter taste, 2- I feel something but cannot identify the taste, and 3-I do not feel the taste. The subjects who reported 2 or 3 were asked to taste higher concentrations of the solution until they expressed perception 1. This was recorded as the threshold for an individual. For individuals who reported a score of 1, at least 1/5th the drug concentration of the actual dose was only allowed for testing the products. A few high concentration API solutions above the individuals perception threshold were made for tasting, and the subjects were subsequently asked to provide a score for each solution (Table 3). The highest concentration of the solution contained CPM equivalent to the dose present in the products tested. The scoring pattern followed was according to modified hedonic scale 0-no taste, 1- tas te something (threshold), 2-slightly bitter, 3-moderately bitter, 4-bitter, and 5-strongly bitter.2.11.2.2. Formulation evaluation and data analysisA washout interval of 12-24 h was allowed after screening the standard solution. The individuals were asked to taste the products (physical mixture or ODF) randomly (blinded) and score the product. The products were placed on the tongue/buccal cavity for 30-40 s, and the subjects were asked to score the bitterness on a scale of 0-5 for each product. Moreover, volunteers were asked to report the time for complete disintegration of the film. Sufficient washout time was allowed between the products, and the volunteers were allowed to drink copious amounts of water after tasting each product. The average of the scores given by all individuals were taken and expressed as the mean standard deviation. The mean scores between the physical mixture and formulation were compared using a student t-test at 95% confidence level, and PResults and Discu ssion3.1. Preparation of hot-melt extruded filmModified starch is very difficult to extrude because of its high glass transition temperature. Thus, there is a need to introduce a plasticizer during extrusion, which could reduce the melt viscosity and increase the free volume of starch chains. For this study, glycerin was used as a plasticizer in different proportions, and it exhibited refined extrudability with significantly lower torque (4.8-7.2 Nm) values than typically encountered. The barrel design was modified with a degassing port to remove excess amount of water vapor from the molten mass. Initial studies without a degassing port demonstrated the front man of bubbles as well as unequal distribution of drug in the film samples.Standard screw configuration (Figure 1) with three mixing zones was utilized for this study. It provided enough shear for dispersive and distributive mixing of the drug and helped get excellent content uniformity in all the extruded film formulations.T he extruded films were stretched using the roll connected to the extruder assembly. This aided in making thin films with uniform thickness, and the roll speed was optimized for steady collection of the film. The combination of processing and formulation parameters helped to manufacture uniform, clear, and very thin films (60-110 m) using melt extruder.3.2. Physiochemical evaluation of filmsTGA is very critical before playing HME because the drug and excipients are exposed to high temperature during the extrusion process, and there are possibilities of drug degradation or thermally-induced chemical reactions or both 16. The TGA results (Figure 2) specified that API, polymer, and excipients were chemically stable in the HME processing temperature range. Lycoat RS 780 demonstrated a loss of weight (33.XRD was used to investigate the physical state of the drug after HME process. The XRD results (Figure 3a 3b) of CPM illustrated prominent peaks at 2 angles of approximately 13, 19, and 20 degrees, while citric acid showed peaks at 2 angles of 18, 19, and 26. The melt-extruded formulation did not show any peak that confirmed the presence of drug in an amorphous form. The reasons behind the complete conversion of drug to an amorphous form were the high shear during extrusion, low drug load, and relatively high amounts of glycerin. The presence of CPM and excipients in an amorphous form aided the flexibility and clear appearance of the film.3.3. Dissolution studiesLycoat RS 780 is a comparatively new modified starch-based polymer, which demonstrated its conditional relation in film coating for tablets and oral film development using solvent casting method 34, 35. Being a non-gelling and highly water-soluble polymer, it provides rapid disintegration and dissolution to formulations. Visual inspection during dissolution demonstrated rapid disintegration of the film when it touched the dissolution media. This characteristic helps in the rapid attempt of action of the f ormulation, because the drug can diffuse from the oral mucosa and reach the systemic circulation 36.During dissolution studies, the formulations rapidly release CPM, and it was attributed to hydrophilic excipients and Biopharmaceutics Classification System (BCS) fork I drug. These films had very low thickness (60-110 m) and higher surface area, which enabled interaction with dissolution media and rapid disintegration following complete dissolution. Dissolution results (Figure 4a 4b) showed 95% drug release in the first 5 min of the dissolution experiment, and at 10-min time points, there was complete release of the drug.3.4. SEM evaluationThe surface morphology was examined by SEM for all film formulations. SEM images did not exhibit CPM crystals, indicating an amorphous nature of drug in formulations (Figure 5). The formulations showed very good-tempered surface at low magnification. This support the smooth texture of film surface, which is one of the esthetic attributes of fil ms. However, at microscopic level, there was high surface area, which helped in the rapid disintegration of the film.3.5. Film characterizationsAs illustrated in Table 2, the film formulations demonstrated excellent D.T of 6-11 s, which was attributed to the thickness of the film and presence of water-soluble materials in the film. The formulations contained water-soluble excipients and APIs such as CPM, citric acid, glycerin, and modified starch. The most crucial parameter for disintegration is the low thickness of the film. As the films had a thickness range of 60-110 m, they aided in the faster disintegration of all film formulations. In addition to the low thickness of the film, the amount of saliva in the oral cavity is very critical for rapid disintegration. The normal flow of saliva in a healthy person is 0.34 mL/min, and it can be increased by the addition of agents that simulate salivary production, including citric, malic, lactic, ascorbic, and tartaric acids 14. citric ac id is the most preferred saliva-stimulating agent, and it was estimated that citric acid could increase salivary flow approximately 5-fold in 2-6% proportion in the formulation 14. With the addition of citric acid, the pH of the films was found to be in the range of 2.9-3.4 and it could contribute in improving rate of salivary flow after administration of formulation which volition aid in rapid disintegration of film product.All the film formulations were tested for their Ts and %E (Table 2). Ideally, the film should have desirable mechanical properties so that it can remain intact during handling and transport. ODFs showed appropriate strength and %E. These excellent mechanical properties were attributed to the presence of glycerol, citric acid, and CPM, which reduced film stiffness via disruption of intermolecular forces of the polymer owing to the accommodation of these compounds between the strands, thereby providing elasticity to the films 37, 38.3.6. In vivo taste evolutionFi rstly, the taste perception of rats was evaluated by administering 0.5 and 1 mg/mL CPM dissolved in distilled water. These results were important to avoid taste variability between animals (Figure 6), and showed that the rats consumed 10 and 14 mL of water in 15 and 30 min, respectively. The results of this study were comparable with those of the study published by Tiwari et.al. Thus, the rate and extent of wasting disease of water were reduced significantly to 3 and 5 mL in 15 and 30 min with the administration of 0.5 mg/mL CPM solution. At a higher concentration of 1 mg/mL, there was notable reduction in consumption of water to 1.6 and 2 mL at 15 and 30 min. Notably, this reduction in consumption of CPM solution despite deprivation of water for 22 h affirmed an annoyance toward CPM. Moreover, aversion behaviors (Figure 7), such as jaw smacking, oral grooming, nose wrinkle, paw wipe, forelimb flail, head shake, paw shakes, and retreating confirmed the dislike of rats toward the d rug solution 39.As illustrated in Figure 6, the rats consumed 10 and 12.5 mL of N 2 solution (5% CPM in the film) in the first 15 and 30 min, and the amount was comparable with the consumption of water. In addition, N7 (10% CPM) exhibited consumption of 4.7 and 8 mL at 15 and 30 min. Furthermore, N9 (7.5% CPM) showed consumption of 6.3 and 10 mL at 15- and 30-min time point. These results indicated that with increasing concentrations of CPM, there was discernible reduction in the consumption of formulation. The rats did not show aversion behavior such as forelimb flail with N2 formulation. However, there was a surge in the aversion behavior response upon increasing the drug concentrations in N7 and N9,.The results of this study were very helpful to understand about the taste of pure drug and formulation. It provided an insight into the taste of products, which helped to screen this formulation for human studies.3.7. Film evaluation by human panelBefore evaluation of taste of the f ormulation, it is very important to understand the taste perception of human volunteers to minimize intra-subject variability. Taste perception study was performed on six healthy human volunteers. Initially, different concentrations of CPM in 2 mL of water were administered to the human subjects. Three subjects had threshold at 0.5 mg and the remaining three subjects reported moderate bitterness at the same concentration. A dose of 2.5 mg demonstrated bitterness in all subjects, and only three volunteers could taste higher concentration of CPM (4 mg, Table 3). This initial evaluation confirmed the appropriateness of the subjects for taste masking study.The results of taste masking evaluation in animal model suggested that formulation N2 with 5% CPM had significant taste masking. Moreover, this formulation had an excellent D.T (7 s) in in vitro studies. Based on t