Effect of Fast-Disintegrating Tablets\u27 Characteristics on the Sublingual Permeability of Atropine Sulfate for the Potential Treatment of Organophosphates Toxicity

Atropine sulfate (AS) fast-disintegrating sublingual tablets (FDSTs) were tested for AS sublingual permeation\u27s feasibility as a potential alternative dosage form to treat organophosphates (OP) toxicity. More than 12,000 OP pesticide toxicity cases were reported in the USA from 2011 to 2014. AS is the recommended antidote for OP toxicity; however, it is only available as an ATROPEN® auto-injector, an IM injection, for self-administration, which is associated with several drawbacks and limitations. Six AS FDST batches were formulated and characterized. Two tablet sizes, group A weighing 150 mg and group B weighing 50 mg, were formulated with three different AS doses: 2 mg (A1 and B1), 4 mg (A2 and B2), and 8 mg (A3 and B3). AS in vitro diffusion and sublingual permeation were investigated in Franz cells using a cellulose membrane and an excised porcine sublingual membrane. The effect of AS load and tablet size on sublingual permeation was also evaluated. All batches passed quality control tests. AS FDSTs\u27 size and AS load had a significant effect on tablet disintegration time and drug dissolution, which significantly impacted AS concentration gradient across the diffusional membrane. Group B FDSTs (smaller tablets) resulted in a significantly higher initial permeation (JAU

Fabrication of Epinephrine Nanoparticles Using Microfluidizer Processor for the Treatment of Anaphylaxis

Purpose. Epinephrine was previously formulated into fast-disintegrating sublingual tablets (AAPS PharmSciTech 2006;7(2):41) and the sublingual bioavailability was established in vivo (J Allergy Clin Immunol 2006;117(2):398-403) for the potential first-aid treatment of anaphylaxis. The purpose of this study was evaluate the feasibility of reducing epinephrine particles size using high shear fluid processor (Microfluidizer) to enhance epinephrine sublingual bioavailability. Methods. (-)-Epinephrine (Epi) and (-)- epinephrine (+)-bitartrate (EpiBit) solubility were tested in various solvents as carriers for shear fluid processing. Epi and EpiBit suspensions were processed using M-110P Microfluidizer at 15 to 30 KPsi for several passes. Particles size was measured before and after processing using NiComp 370 and Mastersizer. Epi and EpiBit stability was monitored visually and by FT-IR. Powder was collected by ART bench top lyophilizer. Results. Water and isopropyl alcohol were selected as carriers for Epi and EpiBit respectively. Epi’s particles size was reduced from 32.9±0.3 μm (mean±SD) to 905.9±82 nm and to 273.9±179 nm after one pass at 15 KPsi and 30 KPsi, respectively. EpiBit’s particles size was reduced from 150.7±5.0 μm to 2.0±0.2 μm after 16 passes at 15 KPsi (1 pass) and 25 KPsi (15 passes). After processing, Epi suspension discolored to pinkish- brownish color and EpiBit did not change color. FT-IR spectrums reflected Epi degradation and confirmed EpiBit stability after processing. The process yield for EpiBit was 68%. Conclusion. Size reduction of Epi was achievable but resulted in its degradation. EpiBit was stable during the shear process and the particle size was reduced to 2 μm

EFFECT OF PARTICLES SIZE ON EPINEPHRINE SUBLINGUAL DIFFUSION FOR THE POTENTIAL FIRST-AID TREATMENT OF ANAPHYLAXIS: IN VITRO AND EX VIVO STUDY

Objective. To evaluate the in vitro and ex vivo diffusion of epinephrine microcrystals (Epi-MC) from our rapidly disintegrating sublingual tablets (RDSTs). Background. Epi 0.3 mg IM injection in the thigh is the drug of choice and the only available dosage form for the treatment of anaphylaxis in community settings. Previously, we showed that Epi 40 mg from RDST is bioequivalent to 0.3 mg IM Injection in our validated rabbit model. We hypothesized that substantial reduction in Epi particles size would significantly enhances its sublingual diffusion. Methods. Epi-MC were prepared by top-down technique using LV-1 Microfluidizer. RDSTs were manufactured by direct compression using our previously developed and published formulation. The in vitro and ex vivo diffusion of Epi 10, 20, and 40 mg RDSTs, and Epi-MC 10, 20 mg RDSTs (n=4) through dialysis and excised sublingual porcine mucosal membranes respectively, were evaluated using Franz cells. Epi 10 mg solution was used as a control. Results. Mean (SD) JAUC0- 90 of diffused Epi, Jmax, and Epi influx (J) from Epi 40 mg RDSTs (484,185±29,656μg/cm2/min, 7,508±569μg/cm2, 234±100μg/cm2/min, respectively) and Epi-MC 20 mg RDSTs (402,852±55,299μg/cm2/min, 6,727±736μg/cm2, 172±50μg/cm2/min, respectively) were not significantly different in vitro (p \u3e 0.05). Mean (SD) JAUC0-90 of diffused Epi, Jmax, and Epi influx (J) from Epi 40 mg RDSTs (264,556±182,820μg/cm2/min, 4,796±2,988μg/cm2, 106±82μg/cm2/min, respectively) and Epi-MC 20 mg RDSTs (211,369±116,025μg/cm2/min, 3,527±1,755μg/cm2, 91±55μg/cm2/min, respectively) were not significantly different ex vivo (p \u3e 0.05). Conclusion. The Epi-MC RDSTs improved Epi diffusion two-fold and have the potential to reduce the bioequivalent dose of sublingually administered Epi by 50%. Grants. This study was funded by the Health Professions Division Grant and the President\u27s Faculty Research & Development Grant, Nova Southeastern University

Advances in ocular drug delivery

Eye drops have long been the primary ocular drug delivery dosage form used to treat ocular disorders ranging from superficial conditions to intravitreal diseases. The ocular anatomical structure and physiological protective mechanisms are one of the most formidable barriers to drug penetration that have significantly reduced the drug\u27s efficacy and target selectivity while sometimes causing ocular tissue damage. There are many new and innovative advances in ocular drug delivery due to better understanding of the structure and function of the eye, the nature of its diseases, and how to overcome or utilize its protective barrier(s), which resulted in increased bioavailability and longer duration of action of the administered drugs, therefore, more effective disease management. We seek in this article to present a comprehensive overview of the basic required knowledge about the barriers for drug delivery to the eye and the major breakthroughs and advances in ocular drug delivery to the anterior, posterior and intravitreal segments of the eye

VALIDATION OF HPLC METHOD AND ULTRA VIOLET DETECTION FOR THE QUANTIFICATION OF ATROPINE SULFATE IN AQUEOUS SAMPLES

Objective. To validate a USP HPLC method for the quantification of Atropine Sulfate (AS) in aqueous samples. Background. AS IM injection is the recommended antidote for management of organophosphate toxicity. We hypothesized that AS sublingual administration can result in similar pharmacokinetic profile as AS IM injection. Methods. An HPLC system and ultra violet detector (Perkin Elmer, MA, USA) were used for the separation and quantification of AS according to USP method for AS injection. AS stock solution (2mg/mL in water) was used to prepare several standard calibration curves ranging from 5¼g/mL to 200¼g/mL at various days to calculate the intra- and inter-assay variability. An increasing injection volumes ranging from 10¼L to 100¼L were injected to test for injection linearity. To test for method and instrument accuracy, the lowest (5¼g/mL) and highest (200¼g/mL) standards were injected 5 times. Mean, standard deviation, and relative standard deviation (RSD%) have been calculated for each validation test. Results. The Calibration curves were linear with a mean (SD) R2 of 0.9966±0.00085 (n=6). The RSD% (n=2) intra-and inter-assay at 5 ¼g/mL were 7.24% and 7.69%, respectively, and at 200 ¼g/mL were 0.076% and 0.83%, respectively. RSD% (n=5) of instrument and method accuracy for the lowest and highest standards (5 ¼g/mL and 200 ¼g/mL) were 3.915% and 0.359%, respectively. The instrument injection was linear with R2 of 0.99998 and the minimum detection limit was 50ng/mL with RSD% (n=5) of 0.86%. Conclusion. A reproducible and sensitive USP HPLC method was validated for the quantification of AS from Aqueous samples. Grants. This study was funded by Saudi Arabian Cultural Mission Gran

DEVELOPMENT AND VALIDATION OF HPLC METHOD WITH PDA DETECTOR FOR SIMULTANEOUS QUANTIFICATION OF CO-ELUTED COMPOUNDS IN PHARMACEUTICAL SAMPLES

Objective. To develop and validate an HPLC method for the separation and quantification of co-eluted dipivefrine, a prodrug, and its active form, epinephrine, in a standardized pharmaceutical sample. Background. The current USP methods for determination of individual compounds are not suitable for their detection when co-eluted. To our knowledge, there is no published analytical method using HPLC with Photo-Diode-Array (PDA) detector for the simultaneous detection of the co-eluent which is required for the development of novel dipivefrine dosage form. Methods. Different strengths of mobile phases were evaluated using methanol and acetonitrile. Several reversedphase C18 and Hilic columns at various pressures and flow rates using Waters Alliance System with PDA detector were evaluated. A 2.0 mg/mL standard stock solution of each compound was prepared in 0.0015N HCL solvent. Standard calibration curves ranging from 20 - 200¼g/mL were injected and plotted for both compounds. The linearity of the injections, the reproducibility of quantification, the minimum detection limit, and the intra and inter assay variation for each compound were tested and validated. Results. Separation was achieved using C18 column 43 (3.9mm X 300mm, 10¼m) with a flow rate of 2.0 ml/min at wavelength of 254 nm at ambient temperature. (More data are being collected and will be presented in the poster). Conclusion. A reliable and sensitive reversed-phase HPLC method was developed and validated for the quantification of dipivefrine and epinephrine from one sample. (Project in progress, more data will be provided to confirm conclusion). Grants. President\u27s Faculty Research & Development Grant FY2014

Formulation and Evaluation of Fast-Disintegrating Sublingual Tablets of Atropine Sulfate: the Effect of Tablet Dimensions and Drug Load on Tablet Characteristics

In this study, we formulated and evaluated the effects of tablet dimensions and drug load on the characteristics of atropine sulfate (AS) fast-disintegrating sublingual tablets (FDSTs). We aim to develop AS FDSTs as an alternative non-invasive and portable dosage form for the emergency treatment of organophosphate (OP) toxicity. AS autoinjector, AtroPen®, is the only self-administered dosage form available as an antidote for-out-of-hospital emergency use, but it is associated with several limitations and drawbacks. Seven FDST formulations of two tablet sizes, 150 mg (A) and 50 mg (B), and of several AS loads, 0 mg (A1, B1), 2 mg (A2, B2), 4 mg (B3), and 8 mg (B4a, B4b), were formulated and manufactured by direct compression. AS FDST characteristics were evaluated using USP and non-USP tests. Results were statistically compared at p \u3c 0.05. All FDSTs passed the USP content uniformity and friability tests, disintegrated and released AS in ≤30 and 60 s. B1 and B2 were significantly harder than A1 and A2. Water uptake of A1 was significantly the highest. However, B1 and B2 had shorter disintegration and wetting times and higher amounts of AS dissolved than did A1 and A2 (p \u3c 0.05). Increasing AS negatively affected FDST tensile strength (p \u3c 0.05 for B4a) and water uptake (p \u3c 0.05 for B3, B4a and B4b), however, without affecting AS dissolution. Formulation of AS up to 16% into smaller FDSTs was successful. Smaller FDSTs were harder and disintegrated more quickly. These AS FDSTS have the potential for further in vivo testing to evaluate their OP antidote potential

In Vivo Evaluation of Taste-Masked Fast-Disintegrating Sublingual Tablets of Epinephrine Microcrystals

In community settings, IM injection of 0.3 mg epinephrine (Epi) using an auto-injector is the drug of choice for treatment of anaphylaxis. Previously, a taste-masking (TM) formulation of fast-disintegrating sublingual tablets (FDSTs) was developed in our lab. Also, Epi was micronized (Epi-MC) successfully and reduced the previously achieved bioequivalent sublingual Epi dose to 0.3 mg IM injection by half using non-taste-masked fast-disintegrating sublingual tablets (TM-FDSTs). Our objective for this study was to evaluate the sublingual absorption of Epi-MC using TM-FDST. These sublingual Epi tablets have potential for out-of-hospital treatment of anaphylaxis and are suitable for human studies. TM-FDSTs containing Epi-MC were manufactured by direct compression. The rate and extent of Epi absorption from our developed 20 mg Epi-MC-TM-FDSTs (n = 5) were evaluated in rabbits and compared to the previous result from 20 mg Epi-MC in non-TM-FDSTs and EpiPen® auto-injector. Blood samples were collected over 1 h, and Epi concentrations were measured using HPLC with electrochemical detection. Mean ± SEM AU