Hydrogen Bonding Principle-Based Molecular Design of a Polymer Excipient and Impacts on Hydrophobic Drug Properties: Molecular Simulation and Experiment.

Although some commercial excipients for improving the solubility of highly crystalline drugs are widely used, they still cannot cover all types of hydrophobic drugs. In this regard, with phenytoin as the target drug, related molecular structures of polymer excipients were designed. The optimal repeating units of NiPAm and HEAm were screened out through quantum mechanical simulation and Monte Carlo simulation methods, and the copolymerization ratio was also determined. Using molecular dynamics simulation technology, it was confirmed that the dispersibility and intermolecular hydrogen bonds of phenytoin in the designed copolymer were better than those in the commercial PVP materials. At the same time, the designed copolymers and solid dispersions were also prepared during the experiment, and the improvement of their solubility was confirmed, which is in accordance with the simulation predictions. The new ideas and simulation technology may be used for drug modification and development.

Preparation and characterization of a hydroxypropyl methylcellulose based wafer for simultaneous delivery of phenytoin and insulin as wound dressing material.

In this study, a novel wafer based on Hydroxypropyl methylcellulose (HPMC) was prepared as a wound dressing for the simultaneous delivery of phenytoin (PT) and insulin; evaluation of the cutaneous wound repair property was performed too. Due to its low water solubility, PT was encapsulated in polymeric micelles (PM) by the film hydration method at different polymer/drug ratios and characterized in terms of particle size (PS), polydispersity index (PdI), zeta potential (ZP), drug loading (DL) %, entrapment efficiency (EE) %, and drug release. Then, the optimized PT loaded PM (PT-PM) was embedded in the wafers prepared from the HPMC polymer, alone or in combination with Carbopol 940 (CB) and xanthan gum (XG). This wafer also contained a fixed amount of insulin (PT-PM-Insulin-wafer). The obtained wafers were evaluated in terms of morphology, water uptake ability, porosity, bioadhesion and hardness features. Finally, the efficacy of the PT-PM-Insulin-wafer was assessed in full-thickness excision wound models. The optimized PT-PM showed the PS of 84.05 ± 1.80 nm, PdI of 0.28 ± 0.22, ZP of -3.38 ± 0.26 mV, DL of 15.63 ± 0.01%, EE of 92.66 ± 0.08%, and the release efficiency of 59.95 ± 0.03%. The results obtained from the XRD studies of PT-PM also demonstrated the transition of the crystalline nature of the PT to the amorphous form, while FTIR studies showed some intermolecular interaction of PT and the Soluplus® copolymer chain. It was also found that the incorporation of XG into HPMC wafers influenced the microstructure, thus increasing the porosity, water uptake ability and bioadhesion. Compared with other groups, the PT-PM-Insulin-wafer group showed the enhancement of wound closure through increasing collagen deposition and re-epithelialization. The present study, therefore, revealed that the PT-PM-Insulin-wafer group might have very promising applications for wound healing.

Potential herb-drug interaction risk of thymoquinone and phenytoin.

Thymoquinone is a main bioactive compound of Nigella sativa L. (N.sativa), which has been used for clinical studies in the treatment of seizures due to its beneficial neuroprotective activity and antiepileptic effects. It has been evidenced that thymoquinone may inhibit the activity of cytochrome P450 2C9 (CYP2C9). However, little is known about the effect of thymoquinone or N.sativa on the pharmacokinetic behavior of phenytoin, a second-line drug widely used in the management of status epilepticus. In this study, we systematically investigated the risk of the potential pharmacokinetic drug interaction between thymoquinone and phenytoin. The inhibitory effect of thymoquinone on phenytoin hydroxylation activity by CYP2C9 was determined using UPLC-MS/MS by measuring the formation rates for p-hydroxyphenytoin (p-HPPH). The potential for drug-interaction between thymoquinone and phenytoin was quantitatively predicted by using in vitro-in vivo extrapolation (IVIVE). Our data demonstrated that thymoquinone displayed effective inhibition against phenytoin hydroxylation activity. Enzyme kinetic studies showed that thymoquinone exerted a competitive inhibition against phenytoin hydroxylation with a Ki value of 4.45 ± 0.51 µM. The quantitative prediction from IVIVE suggested that the co-administration of thymoquinone (>18 mg/day) or thymoquinone-containing herbs (N.sativa > 1 g/day or N.sativa oil >1 g/day) might result in a clinically significant herb-drug interactions. Additional caution should be taken when thymoquinone or thymoquinone-containing herbs are co-administered with phenytoin, which may induce unexpected potential herb-drug interactions via the inhibition of CYP2C9.

Dual-drug delivery of Ag-chitosan nanoparticles and phenytoin via core-shell PVA/PCL electrospun nanofibers.

Dual-drug delivery systems were constructed through coaxial techniques, which were convenient for the model drugs used the present work. This study aimed to fabricate core-shell electrospun nanofibrous membranes displaying simultaneous cell proliferation and antibacterial activity. For that purpose, phenytoin (Ph), a well-known proliferative agent, was loaded into a polycaprolactone (PCL) shell membrane, and as-prepared silver-chitosan nanoparticles (Ag-CS NPs), as biocidal agents, were embedded in a polyvinyl alcohol (PVA) core layer. The morphology, chemical composition, mechanical and thermal properties of the nanofibrous membranes were characterized by FESEM/STEM, FTIR and DSC. The coaxial PVA-Ag CS NPs/PCL-Ph nanofibers (NFs) showed more controlled Ph release than PVA/PCL-Ph NFs. There was notable improvement in the morphology, thermal, mechanical, antibacterial properties and cytobiocompatibility of the fibers upon incorporation of Ph and Ag-CS NPs. The proposed core-shell PVA/PCL NFs represent promising scaffolds for tissue regeneration and wound healing by the effective dual delivery of phenytoin and Ag-CS NPs.

Preparation of Fine-Drugs Layered Spherical Particles with Good Micromeritic and Dissolution Properties through Ultra Cryo-Milling and Mechanical Powder Processing.

The particles of phenytoin (Phe), a poorly water-soluble model drug, were bead-milled alone or co-milled with a hydrophilic waxy additive using an ultra cryo-milling technique in liquid nitrogen (LN2) to improve its dissolution properties. However, the micronized drug particles adhered and aggregated, resulting in poor handling in manufacturing processes such as blending or tableting. To improve the dissolution profile and powder properties of the drug simultaneously, the milled products were secondarily processed together with larger spherical particles by mechanical powder processing. These secondary products were composite particles with a core-shell structure, with fine drug particles adhered and deposited on the core, based on order mixing theory. As a core, three types/sizes of spherical pharmaceutical excipient particles were applied. The resultant composite particles produced much faster release profiles than just milled or co-milled mixtures. In addition, the composite particles showed good micromeritic properties depending on the size of the core particles. These results indicate that the ultra cryo-milling and subsequent dry composite mixing is a potential approach for developing drug particles with improved dissolution.

Massively parallel characterization of CYP2C9 variant enzyme activity and abundance.

CYP2C9 encodes a cytochrome P450 enzyme responsible for metabolizing up to 15% of small molecule drugs, and CYP2C9 variants can alter the safety and efficacy of these therapeutics. In particular, the anti-coagulant warfarin is prescribed to over 15 million people annually and polymorphisms in CYP2C9 can affect individual drug response and lead to an increased risk of hemorrhage. We developed click-seq, a pooled yeast-based activity assay, to test thousands of variants. Using click-seq, we measured the activity of 6,142 missense variants in yeast. We also measured the steady-state cellular abundance of 6,370 missense variants in a human cell line by using variant abundance by massively parallel sequencing (VAMP-seq). These data revealed that almost two-thirds of CYP2C9 variants showed decreased activity and that protein abundance accounted for half of the variation in CYP2C9 function. We also measured activity scores for 319 previously unannotated human variants, many of which may have clinical relevance.

Spectroscopic, density functional theoretical study, molecular docking, and in vitro studies based on anticancer activity studies against A549 lung cancer cell line of diphenylhydantoin adsorbed on AuNPs surface.

The optimized geometry, FT-Raman, FT-IR, surface-enhanced Raman scattering, UV-Vis spectra, frontier molecular orbital analysis, molecular electrostatic potential analysis, and local and global reactivity descriptors of diphenylhydantoin (DPH) and diphenylhydantoin@AuNPs (DPHA) molecule have been investigated with the help of density functional theory method (B3LYP/6-31++G [d,p] together with LANL2DZ) and was compared and analyzed with the corresponding experimental data in order to identify their structural and bonding features responsible for their bioactivity. In-silico (molecular docking) biological activity screening of the molecules together with the in-vitro (SERS and MTT assay) analysis confirms the anticancer activity of DPH and DPHA molecules. The results of the structure-activity studies and bioactivity studies signify that the DPHA molecule is more active than the DPH molecule against lung cancer.

Direct N1-Selective Alkylation of Hydantoins Using Potassium Bases.

Hydantoins, including the antiepileptic drug phenytoin, contain an amide nitrogen and an imide nitrogen, both of which can be alkylated. However, due to the higher acidity of its proton, N3 can be more easily alkylated than N1 under basic conditions. In this study, we explored methods for direct N1-selective methylation of phenytoin and found that conditions using potassium bases [potassium tert-butoxide (tBuOK) and potassium hexamethyldisilazide (KHMDS)] in tetrahydrofuran (THF) gave N1-monomethylated phenytoin in good yield. The applicable scope of this reaction system was found to include various hydantoins and alkyl halides. To explore the function of methylated hydantoins, the effects of a series of methylated phenytoins on P-glycoprotein were examined, but none of methylated products showed inhibitory activity toward rhodamine 123 efflux by P-glycoprotein.

Phenytoin - An anti-seizure drug: Overview of its chemistry, pharmacology and toxicology.

Phenytoin is a long-standing, anti-seizure drug widely used in clinical practice. It has also been evaluated in the context of many other illnesses in addition to its original epilepsy indication. The narrow therapeutic index of phenytoin and its ubiquitous daily use pose a high risk of poisoning. This review article focuses on the chemistry, pharmacokinetics, and toxicology of phenytoin, with a special focus on its mutagenicity, carcinogenicity, and teratogenicity. The side effects on human health associated with phenytoin use are thoroughly described. In particular, DRESS syndrome and cerebellar atrophy are addressed. This review will help in further understanding the benefits phenytoin use in the treatment of epilepsy.

Phenytoin-loaded lipid-core nanocapsules improve the technological properties and in vivo performance of fluidised bed granules.

Lipid-core nanocapsules (LNCs) were recently reported by our group as a suitable binder system to produce fluidised bed granules. However, there is still a lack of knowledge about the influence of using these nanocarriers loaded with a drug on the properties of the granules and their in vivo performance. Therefore, this study was designed to produce innovative fluidised bed granules containing phenytoin-loaded LNCs (LNCPHT) as a strategy to evaluate the influence of the presence of the drug-loaded nanocarriers on their in vitro and in vivo properties. Granules were produced using a mixture of maltodextrin and phenytoin (1:0.004 w/w) as substrate. They were prepared by fluid bed granulation using water or LNCPHT as the liquid binder, affording good yields (73-82%) of granules with low moisture content (<5%). Granules prepared with LNCPHT had larger mean size (122 µm) compared to maltodextrin primary particles (50 µm) due to the formation of solid bridges. Moreover, the use of LNCPHT as the liquid binder improved their powder flow properties. The nanocarriers were recovered after aqueous dispersion (3.00 mg.mL-1 of PHT) with a redispersibility close to 90%. After reconstitution in water, granules containing LNCPHT showed an improved dissolution behaviour compared to those prepared without them. In addition, they showed a higher mucoadhesive effect due to a combined effect of the LNCPHT and maltodextrin in the interactions with porcine intestinal mucosa. Regarding the in vivo studies, granules containing the combination of non-encapsulated PHT and PHT-loaded lipid-core nanocapsules increased the latency to seizures compared to placebo granules, showing effective anticonvulsant effect in mice. In conclusion, the use of drug-loaded nanocapsules as binder is an encouraging approach to produce fluidised bed mucoadhesive granules with improved technological properties and in vivo performance.

Effects of Diluents on Physical and Chemical Stability of Phenytoin and Phenytoin Sodium.

The focus of the present work was to investigate compatibility between commonly used diluents and the drug (salt and acid form of the phenytoin). Lactose monohydrate (LMH), lactitol hydrate (LCT), and mannitol (MNT) were selected based on commercial products information of phenytoin sodium (PS) and phenytoin acid (PHT). Binary mixtures of the drug-diluent were stored at 60°C and 40°C/75% RH. Similarly, two commercial products, namely Product-A and Product-B, were also investigated in in-use stability. Color of PS-LMH changed from white to yellowish-brown and pH dropped by 3.4 units after 4 weeks exposure. FTIR, XRPD, and NIR chemical images indicated disproportionation in PS-LMH and PS-LCT mixtures stored at 40°C/75% RH. Furthermore, PS-LMH also indicated chemical interactions as indicated by distortion of LMH peaks. PHT-diluent mixture did not exhibit any physical and chemical modifications. Product-A changed color, increased weight, dropped pH value, and exhibited disproportionation and chemical reactions. The dissolution of Product-A decreased from 83.3 ± 1.4 to 7.1 ± 4.4% on 8 weeks exposure to 30°C/75% RH. On the other hand, Product-B did not change; however, dissolution decreased by 15%. In conclusion, PS showed disproportionation and chemical reactions with LMH. Therefore, LMH should be avoided in PS formulations.

Three-ply biocompatible pH-responsive nanocarriers based on HNT sandwiched by chitosan/pectin layers for controlled release of phenytoin sodium.

Today, efficient straightforward biocompatible drug carriers have revoluted advanced drug delivery systems. The study aims to investigate the modification of halloysite nanotubes by chitosan (CTS) and pectin (PCN) for forming a new pH-sensitive bionanocomposites via Layer-by-Layer method. The main objective of this study is to improve loading efficiency and control release of phenytoin sodium (PHT) prepared in various pH. The formation of nanocomposite was confirmed through using FTIR, zeta-potential, TG, SEM, XRD, and UV spectroscopy analyses. Based on the obtained results, HNT/CTS/PCN nanocomposite prepared with the molar ratio of 2:1:2 had the best loading capacity (34.6 mg/g) compared with pure HNT (18.3 mg/g). In-vitro studies showed that prepared bionanocomposites had a low release of PHT in the simulated gastric fluid while having a more controlled release in the simulated intestinal fluid. Because of the loading efficiency and controlled release profile, the composites exhibited great potential for the controlled drug delivery of PHT.

Determination of phenytoin in exhaled breath condensate using electromembrane extraction followed by capillary electrophoresis.

Application of hollow fiber-based electromembrane extraction was studied for extraction and quantification of phenytoin from exhaled breath condensate (EBC). Phenytoin is extracted from EBC through a supported liquid membrane consisting of 1-octanol impregnated in the walls of a hollow fiber, and into an alkaline aqueous acceptor solution inside the lumen of the fiber. Under the obtained conditions of electromembrane extraction, that is, the extraction time of 15 min, stirring speed of 750 rpm, donor phase pH at 11.0, acceptor pH at 13.0, and an applied voltage of 15 V across the supported liquid membrane, an enrichment factor of 102-fold correspond to extraction percent of 25.5% was achieved. Good linearity was obtained over the concentration range of 0.001-0.10 µg/mL (r2 = 0.9992). Limits of detection and quantitation were 0.001 and 0.003 µg/mL, respectively. The proposed method was successfully applied to determine phenytoin from EBC samples of patients receiving the drug. No interfering peaks were detected that indicating excellent selectivity of the method. The intra- and interday precisions (RSDs) were less than 14%.

Core-sheath gelatin based electrospun nanofibers for dual delivery release of biomolecules and therapeutics.

Coaxial electrospinning with the ability to use simultaneously two separate solvents provides a promising strategy for drug delivery. Nevertheless, controlled release of hydrophilic and sensitive therapeutics from slow biodegradable polymers is still challenging. To address this gap, we fabricated core-sheath fibers for dual delivery of lysozyme, as a model protein, and phenytoin sodium as a small therapeutic molecule. The sheath was processed by a gelatin solution while the core fibers were fabricated from an aqueous gelatin/PVA solution. Microstructural studies by transmission and scanning electron microscopy reveal the formation of homogeneous core-sheath nanofibers with an outer and inner diameter of 180 ± 48 nm and 106 ± 30 nm, respectively. Thermal gravimetric analysis determines that the mass loss of the core-sheath fibers fall between the mass loss values of individual sheath and core fibers. Swelling studies indicate higher water absorption of the core-sheath mat compared to the separate sheath and core membranes. In vitro drug release studies in Phosphate Buffered Saline (PBS) determine sustained release of the therapeutics from the core-sheath structure. The release trails three stages including non-Fickian diffusion at the early stage followed by the Fickian diffusion mechanism. The present study shows a useful approach to design core-sheath nanofibrous membranes with controlled and programmable drug release profiles.

Nanoemulsions and thermosensitive nanoemulgels of phenytoin and fosphenytoin for intranasal administration: Formulation development and in vitro characterization.

Phenytoin is a low solubility anticonvulsant drug. It has, nonetheless, other possible therapeutic indications, such as neuropathic pain, including trigeminal neuralgia, or wound healing. Its use has decreased due to side effects, but nasal/intranasal administration could significantly increase drug safety and efficacy. The aim of this work was to develop and study nanoemulsions and thermosensitive nanoemulgels of phenytoin and fosphenytoin, in combination, for intranasal administration, with immediate and sustained release profiles. Nanoemulsions were prepared by adding the aqueous phase, containing gelling polymers in the case of nanoemulgels, to emulsion preconcentrates, followed, in the optimized procedure, by premix membrane emulsification. Formulation design and optimization was guided by drug strength, rheological behavior, osmolality, mean droplet size and polydispersity. Fosphenytoin interfered significantly with Carbopol but not with Pluronic's gelation, and allowed to achieve drug strengths equivalent to 22 or 27 mg/g of phenytoin in lead nanoemulsions, and 16.7 mg/g of phenytoin in the lead nanoemulgel. The final selected low viscosity nanoemulsions had an immediate or prolonged fosphenytoin release profile, depending of anhydrous phase proportion (10% or 40%, respectively). The thermosensitive nanoemulgel, with 10% anhydrous phase, showed prolonged drug release. Future studies will establish whether they are more suited for topical effects or therapeutic brain delivery.

Cryo-milling with spherical crystalline cellulose beads: A contamination-free and safety conscious technology.

Crystalline cellulose is a common inactive pharmaceutical additive. If this material can also be used to construct beads for the wet milling of pharmaceutical compounds, it could possibly address issues related to wear and contamination associated with zirconia and polyethylene beads. In this study, the model drug phenytoin was milled with spherical crystalline cellulose (SCC) in liquid nitrogen. The particle size of the milled product was found to be comparable to that obtained using zirconia beads, verifying the feasibility of using SCC beads for this purpose. Using a design of experiment approach, the bead amount, agitation speed, and milling time were all determined to have a significant effect on the milled particle size, giving a D50 value as low as 0.3 µm. No breakage of the SCC beads was observed during the milling process in durability tests under conditions that will degrade spherical D-mannitol beads, showing that this material exhibits sufficient durability. In addition, the variation in elastic modulus between beads was minimal. Because SCC is commercially available and easy to handle, the present wet milling technique is considered to have potential applications to the manufacture of pharmaceuticals on an industrial scale, as it shows sufficient milling capability and durability.

Phenytoin/sildenafil loaded poly(lactic acid) bilayer nanofibrous scaffolds for efficient orthopedics regeneration.

Autologous and synthetic bone grafts showed some limitations during their usage in bone tissue regeneration. This is attributed to several drawbacks such as difficulty of finding a donor in addition to the autoimmune rejection. This study aims to fabricate a well-designed biocompatible double-layered structure of highly porous poly(lactic acid)-based electrospun nanofibers (NFs) as scaffolds for bone tissue regeneration. Poly(lactic acid) was chosen to fabricate the main matrix of the NFs scaffold as it is one of the FDA approved and highly recommended biopolymers for biomedical applications owing to its high biodegradability and biocompatibility Each layer is loaded with a different drug (Phenytoin and Sildenafil) to stimulate bone healing process. The solvents and the parameters of electrospinning were manipulated to produce highly porous structures in order to enhance the in-situ biodegradability of the NFs mats as well as the drug release rate. The produced NFs mats were fully characterized morphologically (SEM), chemically (FTIR), physically (DSC) and physicochemically (biodegradability, swellability, porosity and water vapor permeability) as well as studying the drug release profiles of both drugs. Cytotoxicity of the fabricated NFs was tested using fibroblast cells to detect their biocompatibility. Cell adhesion and proliferation were examined using SEM before using the NFs as scaffolds in mice animal model. The efficiency of the developed NFs scaffolds in healing bone fractures was assessed after 14 and 28 days through visual inspection, SEM investigation and bone mineral density assessment. Finally, sections from the bone fracture sites were isolated for histopathological examination. The study revealed the efficiency of the drugs-loaded NFs in enhancing cell adherence, cell proliferation, angiogenesis formation and finally tissue restoration of bone fractures.

Cryo-milling using a spherical sugar: Contamination-free media milling technology.

Milling beads experience wear upon repeated use. And milling beads made of material that is safe when ingested have not yet been developed. The present report describes the development and characteristics of spherical d-mannitol (SDM) beads, which would be safe when ingested. The model drug phenytoin was dispersed in liquid nitrogen along with SDM and the materials were agitated at high speed. The effects of the amount of beads, agitation speed, and milling time on phenytoin particle size, yield, and bead fractures were investigated using a central composite experimental design. The diameter of milled phenytoin particles decreased significantly as the amount of SDM beads and agitation speed increased. In contrast, no difference was found in the diameter with milling time. Although the fractured SDM ratio increased slightly at higher agitation speeds, the SDM was not broken and was durable enough for milling. This milling technique was applicable not only to phenytoin but also to other drug substances. Bead durability and applicability indicated that SDM can be used as wet milling beads that are considered safe for use if ingested.

Novel contamination-free wet milling technique using ice beads for poorly water-soluble compounds.

The aim of this study is to establish a contamination-free milling method using ice beads instead of conventional hard beads such as metal or ceramics. Ice beads, which melt after the milling process to form water, would solve the contamination issue attributed to bead breakage or abrasion. The technique/method for preparing spherical ice beads of mono-dispersed size ranging from 150 to 3000 µm was newly developed. An oscillation beads milling apparatus was used for pulverization. In the initial stages of ice beads milling, the process is dry, but as time passes, the surface of the ice beads begins to melt, resulting in a transition to wet beads milling. It was found that ice beads are an effective milling media for beads milling, and that milling efficiency is strongly affected by the temperature of the coolant, with the peak efficiency occurring when the temperature was set to -2 °C and ice beads around 1500 µm in diameter were used. The spray-dried powder obtained from suspension after ice beads milling had dissolution improvement equivalent to that obtained after zirconia beads milling, resulting from its spontaneous rapid dispersion into nanosuspension.

Quality and In-Use Stability Comparison of Brand and Generics of Extended-Release Phenytoin Sodium Capsules.

The objective of the present study was to understand quality of brand and generic products of phenytoin sodium by in vitro methods. Three commercial products were selected for the study, 1 brand and 2 generics (product-A, product-B, and product-C). Products were repacked in pharmacy vials and stored for 12 weeks at 30°C/75% RH to simulate in-use conditions. The products were examined visually and microscopically for morphologic changes, spectroscopic and diffractometric methods for chemical changes, and dissolution, assay, and impurities for performance evaluation. Capsules content of the product-A turned yellowish to dark orange color from initial white powder, which indicated a possible chemical interaction between lactose and the drug in addition to disproportionation. This was supported by pH, microscopic, spectroscopic, and X-ray diffraction data. Product-A failed to meet United States Pharmacopoeia dissolution specification of 75% in 120 min after 2-weeks whereas product-B and product-C failed at 6-weeks of in-use stability conditions exposure. Furthermore, product-A also failed to meet United States pharmacopoeia assay and impurities specifications in 12 weeks in-use period. In summary, this study indicated salt disproportionation, chemical interactions, and phase transformations of drug and excipients in the commercial products of phenytoin sodium, which may affect the clinical performance of the product.