Formulation Screening Research Articles

Engineering, on-demand manufacturing, and scaling-up of polymeric nanocapsules.

Polymeric nanocapsules are versatile delivery systems with the capacity to load lipophilic drugs in their oily nucleus and hydrophilic drugs in their polymeric shell. The objective of this work was to expand the technological possibilities to prepare customized nanocapsules. First, we adapted the solvent displacement technique to modulate the particle size of the resulting nanocapsules in the 50–500 nm range. We also produced nanosystems with a shell made of one or multiple polymer layers i.e. chitosan, dextran sulphate, hyaluronate, chondroitin sulphate, and alginate. In addition, we identified the conditions to translate the process into a miniaturized high‐throughput tailor‐made fabrication that enables massive screening of formulations. Finally, the production of the nanocapsules was scaled‐up both in a batch production, and also using microfluidics. The versatility of the properties of these nanocapsules and their fabrication technologies is expected to propel their advance from bench to clinic.

Poiretia latifolia essential oil as a promising antifungal and anti-inflammatory agent: Chemical composition, biological screening, and development of a nanoemulsion formulation

Poiretia latifolia is a Brazilian plant species used in traditional medicine for a wide range of ailments. The objective of this study was to evaluate the antioxidant potential, anti-inflammatory activity, and antifungal effect of P. latifolia essential oil and its major compounds. In addition, a nanoemulsified system was developed for its application. Aerial parts of the plant, collected at different stages of development, were hydrodistilled, and the chemical composition of the oil was determined by gas chromatography–mass spectrometry (GC–MS). Limonene, trans-dihydrocarvone, and carvone were identified as major compounds. Antioxidant potential was evaluated by three different methods: TBARS, β-carotene/linoleic acid, and DPPH. P. latifolia oil showed antioxidant activity (0.508 mg/mL) in lipophilic medium, as evidenced by the β-carotene/linoleic acid assay. The anti-chemotactic effect of the oil was evaluated by inhibition of neutrophil migration (0.001–10 μg/mL): P. latifolia oil was able to inhibit up to 94.7% of migration. Antifungal activity was evaluated against yeasts (Candida spp.) and dermatophytic filamentous fungi (Microsporum spp. and Trichophyton spp.). P. latifolia oil showed antifungal activity against Microsporum canis, and a mixture of carvone and limonene was effective against the dermatophytes Trichophyton rubrum and Trichophyton mentagrophytes. Nanoemulsions were successfully developed. P. latifolia oil has the potential to be used as an antidermatophytic and anti-inflammatory agent.

Chemical admixtures for the optimization of the AAC production

AbstractThe use of chemical admixtures for the improvement of the manufacturing process, as well as for the quality enhancement of elements and buildings, is a regular practice in standard high consistency concrete, both in ready‐mix and in precast applications, but is still quite limited in the production of autoclaved aerated concrete (ACC). This is partially due to the difficulty of replicating the manufacturing process in the laboratory, which includes expansion and setting in the precuring stage, followed by the autoclave treatment at elevated temperature and pressure. This work presents a structured approach to the design of specific chemical admixtures for ACC, which started with the identification of suitable laboratory equipment, and test methods for the simulation of the production process of AAC. This was then expanded with the screening and selection of different chemical formulations, to end with the development of chemical admixtures tailored for the ACC. The results show that, by using these specific admixtures, the producers of AAC can accelerate the production process, thereby increasing the output of the plant; alternatively, when the increase in productivity is not an objective, the same admixtures can be employed to achieve a significant optimization in the composition of the mix.

Drying Technologies for the Stability and Bioavailability of Biopharmaceuticals.

Solid dosage forms of biopharmaceuticals such as therapeutic proteins could provide enhanced bioavailability, improved storage stability, as well as expanded alternatives to parenteral administration. Although numerous drying methods have been used for preparing dried protein powders, choosing a suitable drying technique remains a challenge. In this review, the most frequent drying methods, such as freeze drying, spray drying, spray freeze drying, and supercritical fluid drying, for improving the stability and bioavailability of therapeutic proteins, are discussed. These technologies can prepare protein formulations for different applications as they produce particles with different sizes and morphologies. Proper drying methods are chosen, and the critical process parameters are optimized based on the proposed route of drug administration and the required pharmacokinetics. In an optimized drying procedure, the screening of formulations according to their protein properties is performed to prepare a stable protein formulation for various delivery systems, including pulmonary, nasal, and sustained-release applications.

Quantitative screening of the pharmaceutical ingredient for the rapid identification of substandard and falsified medicines using reflectance infrared spectroscopy.

The World Health Organization suggests that approximately 10% of medicines worldwide are either falsified or substandard with higher figures in low and middle income countries. Such poor quality medicines can seriously harm patients and pose a threat to the economy worldwide. This study investigates attenuated total reflectance-fourier transform infrared (ATR-FTIR) spectroscopy as a simple and rapid method for determination of drug content in tablet dosage forms. Paracetamol was used as the model pharmaceutical ingredient. Spectra of standard mixtures of paracetamol with different excipients formed the basis for multivariate PLS based quantitative analysis of simulated tablet content using different selected infrared absorbance bands. Calibration methods using ATR-FTIR were compared with the ATR-FTIR and conventional ultraviolet spectroscopic analyses of real tablet samples and showed that the paracetamol/microcrystalline cellulose mixtures gave optimum results for all spectral bands tested. The quantitative data for band 1524–1493cm-1 was linear (R2 ˃ 0.98; LOQ ≥ 10%w/w tablet). Global examples of paracetamol tablets were tested using this protocol and 12% of the tablet samples examined was identified as substandard. Each sample analysis was completed in just a few minutes. ATR-FTIR can therefore be used in the rapid screening of tablet formulations. The simplicity of the proposed method makes it appropriate for use in low and middle income countries where analytical facilities are not available.

Catalent acquires drug services firm

The drug delivery and development services firm Catalent has agreed to acquire Juniper Pharmaceuticals for $133 million. Juniper’s pharma services division offers drug industry customers solid-state and preclinical formulation screening as well as bioavailability enhancement technologies. Catalent says Juniper’s Nottingham, England, facility will be a European complement to its drug development centers in Somerset, N.J. and San Diego.

In vitro methods to assess drug precipitation in the fasted small intestine - a PEARRL review.

Drug precipitation invivo poses a significant challenge for the pharmaceutical industry. During the drug development process, the impact of drug supersaturation or precipitation on the invivo behaviour of drug products is evaluated with invitro techniques. This review focuses on the small and full scale invitro methods to assess drug precipitation in the fasted small intestine. Many methods have been developed in an attempt to evaluate drug precipitation in the fasted state, with varying degrees of complexity and scale. In early stages of drug development, when drug quantities are typically limited, small-scale tests facilitate an early evaluation of the potential precipitation risk invivo and allow rapid screening of prototype formulations. At later stages of formulation development, full-scale methods are necessary to predict the behaviour of formulations at clinically relevant doses. Multicompartment models allow the evaluation of drug precipitation after transfer from stomach to the upper small intestine. Optimisation of available biopharmaceutics tools for evaluating precipitation in the fasted small intestine is crucial for accelerating the development of novel breakthrough medicines and reducing the development costs. Despite the progress from compendial quality control dissolution methods, further work is required to validate the usefulness of proposed setups and to increase their biorelevance, particularly in simulating the absorption of drug along the intestinal lumen. Coupling results from invitro testing with physiologically based pharmacokinetic modelling holds significant promise and requires further evaluation.

Improved Fluorescence Methods for High-Throughput Protein Formulation Screening.

The goal of protein formulation development is to identify optimal conditions for long-term storage. Certain commercial conditions (e.g., high protein concentration or turbid adjuvanted samples) impart additional challenges to biophysical characterization. Formulation screening studies for such conditions are usually performed using a simplified format in which the target protein is studied at a low concentration in a clear solution. The failure of study conditions to model the actual formulation environment may cause a loss of ability to identify the optimal condition for target proteins in their final commercial formulations. In this study, we utilized a steady-state/lifetime fluorescence-based, high-throughput platform to develop a general workflow for direct formulation optimization under analytically challenging but commercially relevant conditions. A high-concentration monoclonal antibody (mAb) and an Alhydrogel-adjuvanted antigen were investigated. A large discrepancy in screening results was observed for both proteins under these two different conditions (simplified and commercially relevant). This study demonstrates the feasibility of using a steady-state/lifetime fluorescence plate reader for direct optimization of challenging formulation conditions and highlights the importance of performing formulation optimization under commercially relevant conditions.

Design and Optimization of Hydrophilic Matrix-based Sustained Release Felodipine Tablets

Felodipine is a calcium channel blocker used for hypertensive and unstable angina treatments. The sustained release formulations of felodipine have advantages of achieving good therapeutic effects, increasing the bioavailability, decreasing dosing times per day and reducing side effects.The aim of our study was to study the formulation screening, then use an experiment design for formulating a hydrophilic matrix sustained release tablet of felodipine. Methods: The optimization process had the influences of the chosen excipients (including HPMC E4M, HPMC E15LV) on the drug release. Three dependent variables were percentages of released felodipine at the sampling times 2h, 6h, 10h (Y2, Y6, Y10, respectively). Results: The release profile from the optimized formula almost met the predicted release profile and was similar to reference tablets. The kinetics of drug release the optimized tablets and reference tablets were also followed the Korsmeyer – Peppas model.Conclusion: The optimized formula was obtained, and the in vitro release profile was similar to the predicted profile and reference tablets.

High‐Throughput Miniaturized Screening of Nanoparticle Formation via Inkjet Printing

AbstractThe self‐assembly of specific polymers into well‐defined nanoparticles (NPs) is of great interest to the pharmaceutical industry as the resultant materials can act as drug delivery vehicles. In this work, a high‐throughput method to screen the ability of polymers to self‐assemble into NPs using a picoliter inkjet printer is presented. By dispensing polymer solutions in dimethyl sulfoxide (DMSO) from the printer into the wells of a 96‐well plate, containing water as an antisolvent, 50 suspensions are screened for nanoparticle formation rapidly using only nanoliters to microliters. A variety of polymer classes are used and in situ characterization of the submicroliter nanosuspensions shows that the particle size distributions match those of nanoparticles made from bulk suspensions. Dispensing organic polymer solutions into well plates via the printer is thus shown to be a reproducible and fast method for screening nanoparticle formation which uses two to three orders of magnitude less material than conventional techniques. Finally, a pilot study for a high‐throughput pipeline of nanoparticle production, physical property characterization, and cytocompatibility demonstrates the feasibility of the printing approach for screening of nanodrug delivery formulations. Nanoparticles are produced in the well plates, characterized for size and evaluated for effects on metabolic activity of lung cancer cells.

Application of an In Vitro Dissolution/Permeation System to Early Screening of Oral Formulations of Poorly Soluble, Weakly Basic Drugs Containing an Acidic pH-Modifier

This study aimed to evaluate the usefulness of the dissolution/permeation system (D/P system) as an in vitro tool for early screening of oral formulations of weakly basic drugs containing an acidic pH-modifier. Dipyridamole, having a prominent pH-dependent solubility, was used as a model drug, and various granules containing different amounts of fumaric acid were prepared. Prepared granules were administered orally to hypochlorhydria model rats. It was confirmed that fumaric acid improved the absorption of dipyridamole depending on its amount in the granules. Separately, dissolution and permeation of dipyridamole were observed in vitro in the D/P system. When using a medium with a low buffer capacity which mimicked the human intestinal fluid, rank order of the permeated amount of dipyridamole from various granules in the D/P system did not correlate with its absorption in hypochlorhydric rats. In contrast, when applying a medium with high buffer capacity, the permeated amount in the D/P system well reflected the effects of fumaric acid on the in vivo absorption of dipyridamole. In conclusion, by setting appropriate experimental protocols according to the properties of test compounds and formulations, D/P system can be a potent in vitro tool to predict in vivo performance of oral formulations.

Design of a Small-Scale Multi-Inlet Vortex Mixer for Scalable Nanoparticle Production and Application to the Encapsulation of Biologics by Inverse Flash NanoPrecipitation

Flash NanoPrecipitation is a scalable approach to generate polymeric nanoparticles using rapid micromixing in specially designed geometries such as a confined impinging jets mixer or a Multi-Inlet Vortex Mixer (MIVM). A major limitation of formulation screening using the MIVM is that a single run requires tens of milligrams of the therapeutic. To overcome this, we have developed a scaled-down version of the MIVM, requiring as little as 0.2 mg of therapeutic, for formulation screening. The redesigned mixer can then be attached to pumps for scale-up of the identified formulation. It was shown that Reynolds number allowed accurate scaling between the 2 MIVM designs. The utility of the small-scale MIVM for formulation development was demonstrated through the encapsulation of a number of hydrophilic macromolecules using inverse Flash NanoPrecipitation with target loadings as high as 50% by mass.

UV-vis Imaging of Piroxicam Supersaturation, Precipitation, and Dissolution in a Flow-Through Setup.

Evaluation of drug precipitation is important in order to address challenges regarding low and variable bioavailability of poorly water-soluble drugs, to assess potential risk of patient safety with infusion therapy, and to explore injectable in situ suspension-forming drug delivery systems. Generally, drug precipitation is assessed in vitro through solution concentration analysis methods. Dual-wavelength UV-vis imaging is a novel imaging technique that may provide an opportunity for simultaneously monitoring changes in both solution and solid phases during precipitation. In the present study, a multimodal approach integrating UV-vis imaging, light microscopy, and Raman spectroscopy was developed for characterization of piroxicam supersaturation, precipitation, and dissolution in a flow-through setup. A solution of piroxicam dissolved in 1-methyl-2-pyrrolidinone was injected into a flowing aqueous environment (pH 7.4), causing piroxicam to precipitate. Imaging at 405 and 280 nm monitored piroxicam concentration distributions during precipitation and revealed different supersaturation levels dependent on the initial concentration of the piroxicam solution. The combination with imaging at 525 nm, light microscopy, and Raman spectroscopy measurements demonstrated concentration-dependent precipitation and the formation, growth, and dissolution of individual particles. Results emphasize the importance of the specific hydrodynamic conditions on the piroxicam precipitation. The approach used may facilitate comprehensive understanding of drug precipitation and dissolution processes and may be developed further into a basic tool for formulation screening and development.

High-Throughput In-Use and Stress Size Stability Screening of Protein Therapeutics Using Algorithm-Driven Dynamic Light Scattering

Stability of therapeutic proteins (TPs) is a critical quality attribute that impacts both safety and efficacy of the drug. Size stability is routinely performed during and after biomanufacturing. Dynamic light scattering (DLS) is a commonly used technique to characterize hydrodynamic size of the TPs. Herein, we have developed a novel method to evaluate in-use and thermal stress stability of TPs using algorithm-driven high-throughput DLS. Five marketed TPs were tested under the guidance of customized algorithms. The TPs were evaluated at relevant temperature conditions as well as under dilution and thermal stress for size stability. We found that the TPs were stable under the in-use conditions tested; however, sample loss due to evaporation can lead to large protein aggregates. A combined assessment of autocorrelation function and photos of sample well could be useful in formulation screening. Dilution of TPs also has an impact on the hydrodynamic size. Thermal stress experiments showed the importance of using different data processing methods to access size distribution. Polydispersity index was useful in evaluating sample heterogeneity. Herein, we show that algorithm-driven high-throughput DLS can provide additional supportive information during and after biomanufacturing and the potential to be used in a quality control environment.

Cryopreservation of cultured mantle cells of Paphia malabarica for perennial availability

Laboratory friendly, cryopreservation procedures with respect to cryopreservation formulations and cryopreservation temperatures were attempted, in the present study to ensure perennial availability of cultured mantle cells of bivalve (Paphia malabarica). Screening of cryopreservative formulations with different concentrations of DMSO, Propylene glycol and Glycerol was carried out for cryopreservation of freshly dissociated cells of Paphia malabarica. Out of these cryopreservative formulations, 10% DMSO, 10% Propylene glycol and 15% Glycerol were selected for cryopreservation of the mantle cells pooled from 1-day old primary culture and cell line after 3 passages at the end of different cryopreservation periods. Cryopreservative formulation with 15% glycerol, served as a best cryoprotectant for the cryopreservation of cells sourced from freshly dissociated cells as well as from primary cultures and cell cultures after three passages of mantle cells of Paphia malabarica, retaining metabolic activity of resurrected cells. Both, cell cultures established from uncryopreserved cells as well as cryopreserved cells showed similar alkaline phosphatase and carbonic anhydrase activities thus indicating retention of their biomineralization capacity even after cryopreservation at low and ultralow temperatures.

Computer-Aided Formulation Design for a Highly Soluble Lutein-Cyclodextrin Multiple-Component Delivery System.

Cyclodextrin (CD) complexation is widely used for the solubilization of poorly soluble drugs in the pharmaceutical industry. Current research was to develop a highly soluble lutein-cyclodextrin multiple-component delivery system (lutein-CD-MCDS) by combined modeling and experimental approaches. Both phase solubility diagram and molecular dynamics (MD) simulation results revealed that the interactions between lutein and CDs were very weak, which confirmed the insignificant solubility improvement of lutein-CD binary system. On the basis of theoretical calculation and preliminary CD studies, lutein-CD-MCDS was developed with over 400-fold solubility improvement after formulation screening. MD simulation indicated that the auxiliary polymers of TWEEN 80 and poloxamer 188 in the lutein-CD-MCDS introduced bridged interaction between lutein and γ-CD to increase the solubility, dissolution rate, and stability of the complex. The lutein-CD-MCDS was characterized by in vitro dissolution test, differential scanning colorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), and powder X-ray diffraction (PXRD). Moreover, lutein-CD-MCDS had significantly higher uptake in Caco-2 cells than free lutein. The relative bioavailability of the lutein-CD-MCDS increased to 6.6-fold compared to pure lutein, and to 1.2-fold compared with commercial lutein soft capsules. In conclusion, the highly soluble lutein-CD-MCDS with significant improvement in both the solubility and bioavailability was developed and characterized by combined modeling and experimental approaches. Our research indicates that computer-aided formulation design is a promising approach for future formulation development.

An Unexpected Degradation Pathway of a 1,2,4-Triazolo[4,3-a]pyridine Derivative: The Formation of 2 Cationic Pseudodimers of an 11β-Hydroxysteroid Dehydrogenase Type 1 Inhibitor Drug Candidate in a Stressed Capsule Formulation

Degradation of an active pharmaceutical ingredient (API), a 2-(3-(1-(4-chlorophenyl)cyclopropyl)-[1,2,4]triazolo[4,3-a]pyridin-8-yl)propan-2-ol hydrochloride salt, was observed in a capsule formulation stressed at 50°C or 40°C/75% relative humidity conditions for 1 month. Two unknown degradants were identified as cationic pseudodimers of the API via accurate mass liquid chromatography-mass spectrometry and 1- and 2-dimensional NMR analyses. A plausible degradation pathway of the API was postulated which led to the identification of 2 key N-oxide degradants in the stressed capsule formulation at trace levels. It was hypothesized that the N-oxide degradants could be protonated and undergo further transformation so as to react with another API free base to form pseudodimeric N-oxide intermediates, followed by protonation/dehydration to yield the cationic pseudodimers of the API. The proposed degradation pathway was further supported by formulation screening studies: (1) the removal of magnesium stearate (base/lubricant) from the formulation to reduce the formation of API free base, which is susceptible to oxidation to form N-oxides; (2) the replacement of API hydrochloride salt by its free base form to eliminate the proton source for protonation of the N-oxides so as to prevent their further transformation; and (3) the addition of anti-oxidants to minimize the oxidation of API free base to N-oxides.

The Na3V2(PO4)2F3/Carbon Na-Ion Battery: Its Performance Understanding as Deduced from Differential Voltage Analysis

The Na3V2(PO4)2F3/Carbon Na-ion battery technology is appealing because of its high energy and power density, hence the need to assess its viability under practical operation. Here we provide a comprehensive understanding of its storage and cycling performances in various electrolytes and at different temperatures via a through differential voltage (dV/dQ) analysis of the data obtained from coin and pouch cells. The cells show excellent long-term cycling performance (capacity decay less than 0.020 mAh g−1 per cycle at 1C rate whatever the electrolytes), but suffer at 55°C from copious self-discharge stemming from either reduction or oxidization of electrolytes. We could deduce, from monitoring side reactions rate as a function of temperature, the activation energies of 34 and 29 kJ mol−1 for the parasitic reactions proceeding at the anode and cathode side, respectively. We hope our analysis protocol to be widely implemented for the fundamental investigation of side reactions between electrode and electrolyte interface in Na-ion battery; a must for optimizing battery performances via the screening of various electrolyte formulations.

Seroprevalence of Hepatitis B Virus Surface Antigen among Butchers and Slaughtered Cows in Kano Metropolis, Nigeria

Hepatitis B Virus is one of the most important human pathogens which present a significant threat to public health. The objectives of this study are to establish evidence of existence of Hepatitis B Virus (HBV) in cows and prevalence of Hepatitis B Surface Antigen (HBsAg) among butchers. A total of 138 human samples (including slaughter house workers and meat sellers) and 138 slaughtered cows’ samples were collected for detection of HBsAg using ELISA. A questionnaire was administered to collect relevant information regarding risk factors. Screened result showed prevalence rate of 11.6% (p< 0.05) among butchers and 10.9% prevalence was detected in slaughtered cows (p<0.05), these were however not statistically significantly different. Occupational practices including hand cuts and sharing of knives were seen to be more common among slaughter house workers than the meat sellers thereby increasing their exposure to hepatitis B virus infection. It is concluded that both cows and butchers in Kano metropolis serve as reservoir for Hepatitis B Virus and hence there is need for urgent policy formulation (screening and use of protective clothing) in slaughter houses in Nigeria.

Mucus-PVPA (mucus Phospholipid Vesicle-based Permeation Assay): An artificial permeability tool for drug screening and formulation development

The mucus layer covering all mucosal surfaces in our body is the first barrier encountered by drugs before their potential absorption through epithelial tissues, and could thus affect the drugs’ permeability and their effectiveness. Therefore, it is of key importance to have in vitro permeability models that can mimic this specific environment. For this purpose, the novel mucus phospholipid vesicle-based permeation assay (mucus-PVPA) has been developed and used for permeability screening of drugs and formulations. The model proved to be stable under the chosen conditions and demonstrated the ability to discriminate between compounds with different chemical structures and properties. Overall, a decrease in drug permeability was found in the presence of mucus on top of the PVPA barriers, as expected. Moreover, mucoadhesive (chitosan-coated) and mucopenetrating (PEGylated) liposomes were investigated in the newly developed model. The mucus-PVPA was able to distinguish between the different liposomal formulations, confirming the penetration potential of the tested formulations and the related drug permeability. The mucus-PVPA model appears to be a promising in vitro tool able to mimic the environment of mucosal tissues, and could therefore be used for further drug permeability screening and formulation development.