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Natural products are promising drug candidates with various pharmacological effects. However, they can be difficult to use due to poor bioavailability or low stability. In this experiment, while developing topical formulations containing 0.1% Agrimonia pilosa extract, a simple and rapid method of analyzing flavonoid glucuronides, which are representative ingredients, was developed and validated, and the physicochemical properties and stability of flavonoid glucuronides were observed. As a result, an optimized cream formulation was developed. The oil phase comprised tween 60, liquid paraffin, propylene glycol, cetanol, stearyl alcohol, span 60, benzyl alcohol, and A. pilosa extracts, and the water phase comprised water and citric acid hydrate. Then, physicochemical and microbial stability tests of the formulation were conducted under long-term (12 months) and accelerated conditions (6 months). It was thus confirmed that both physicochemical and biological properties were stable during the test period. Consequently, an optimized formulation for the extract was developed and stability was confirmed.

Previously our laboratory first reported that dropping of freeze-dried monoclonal antibody (mAb) formulations could cause protein degradation and aggregation (J Pharm Sci, 2021, 1625). In this manuscript, we evaluated effects of secondary package on stability of several freeze-dried biopharmaceutical formulations during dropping. The degradation of mAb-Y during dropping with different secondary packages was determined by the sensitive particle analyzing techniques micro-flow imaging (MFI) and dynamic light scattering (DLS). Electron paramagnetic resonance (EPR) was used to detect free radicals after repeated dropping in different secondary packages. The amount of free radicals and SbVPs was correlated to the sample temperature as well as the secondary package during dropping. Our observations suggest that secondary packaging has significant effect on freeze-dried biopharmaceutical stability during dropping and therefore should be thoroughly screened and optimized to assure high product quality even for the presumed highly stable freeze-dried biopharmaceuticals.

Fluconazole is an important drug in the treatment of cutaneous and systemic mycoses. The Hospital de Clínicas de Porto Alegre performs a derivation of fluconazole capsules to obtain an oral liquid formulation that is easily administered and whose dose can be adjusted. In order to replace the derivation for a formulation produced from an active pharmaceutical ingredient, this study sought to develop a liquid oral formulation, evaluate its physical chemical and microbiological stability and demonstrate suitability of the analytical method for the formulation assay. Seven different formulations of pharmaceutical suspension form were produced and evaluated for pH, viscosity, sedimentation volume and assay. The analytical method by High Performance Liquid Chromatography was demonstrated. Two most promising formulations were manipulated in the Farmácia Semi-Industrial do Hospital de Clínicas de Porto Alegre and stored in amber PET bottles under three different conditions: room temperature, under refrigeration (2 to 8 ºC) and in an oven (40 ° C). Samples were collected after 0, 7 and 14 days to evaluate physical-chemical stability, assay, pH and macroscopic aspects. Samples were collected after 0 and 21 days to evaluate microbiological stability. It was possible to demonstrate stability for one of the formulations for a 14-day period. Throughout the study, the chosen formulation presented adequate quantification of fluconazole, constant pH, no organoleptic changes and no microbial growth. The results suggest the incorporation of a new formulation for fluconazole to the Farmacia Semi-Industrial portfolio).

The food and beverage industry often need to encapsulate hydrophobic functional ingredients in their products, including colors, flavors, lipids, nutraceuticals preservatives, and vitamins. Encapsulation can improve the handling, water-dispersibility, chemically stability, and efficacy of these functional ingredients. In this review article, we focus on the design of nanoemulsion-based delivery systems to encapsulate, protect, and deliver non-polar bioactive agents, such as vitamin A, D and E, β-carotene, lycopene, lutein, curcumin, resveratrol, and coenzyme Q10. Initially, the challenges associated with incorporating these different bioactives into foods are highlighted. The relative merits and drawbacks of different nanoemulsion fabrication methods are then discussed. Finally, examples of the application of nanoemulsions for improving the stability and bioavailability of various kinds of hydrophobic vitamins and nutraceuticals are provided.

Objectives: The aim of current research was to formulate limonene selfnanoemulsified delivery system (SNEDS) by spontaneous emulsification method. Methods: The optimization was carried out through the construction of a pseudo-ternary phase diagram. Limonene-based self- nanoemulsifying system was optimized by evaluating its droplet characteristic; namely; size, polydispersity, zeta potential, in addition, its morphology was assessed through the use of transmission electron microscopy. Moreover, the formulation stability under different storage conditions for three months was examined. Results: The obtained results showed that the optimized limonene-based self-nanoemulsifying system was characterized by a small droplet size, low polydispersity index, high percentage transmittance and optimal zeta potential with uniform spherical droplets. The selected formulation with 50% w/w limonene, 40% w/w Tween 80 and 10% w/w propylene glycol had a droplet size of 113.3±1.18nm with bluish transparent appearance and a zeta potential value of -19.13±0.38 mV. The developed formula was stable against pH change. The stored limonenebased SNEDS showed acceptable stability at 4°C and 0°C compared to 25°C. Conclusion: The formulated self-nanoemulsifying system showed an improved aqueous dispersibility, patient acceptability and stability of limonene, representing a promising carrier for lipophilic drugs.

Formulation screening for biotherapeutics can cover a vast array of excipients and stress conditions. These studies consume quantities of limited material and, with higher concentrated therapeutics, more material is needed. Here, we evaluate the use of crystal zenith (CZ) microtiter plates in conjunction with FluoroTec-coated butyl rubber mats as a small-volume, high-throughput system for formulation stability studies. The system was studied for evaporation, edge effects, and stability with comparisons to type 1 glass and CZ vials for multiple antibodies and formulations. Evaporation was minimal at 4°C and could be reduced at elevated temperatures using sealed, mylar bags. Edge effects were not observed until 12 weeks at 40°C. The overall stability ranking as measured by the rate of change in high molecular weight and percent main peak species was comparable across both vials and plates at 4°C and 40°C out to 12 weeks. Product quality attributes as measured by the multi-attribute method were also comparable across all containers for each molecule formulation. A potential difference was measured for subvisible particle analysis, with the plates measuring lower particle counts than the vials. Overall, CZ plates are a viable alternative to traditional vials for small-volume, high-throughput formulation stability screening studies.

Meglumine antimoniate (MA) is a pentavalent antimony (SbV) drug recommended for the treatment of leishmaniasis. It is known that the trivalent antimony (SbIII) present as a residue in MA contributes to the drug side effects. In this article, multivariate optimization was used in the synthesis of MA in order to obtain a drug with low levels of SbIII. Four variables (source of antimony, temperature, water volume and pH) were preliminarily evaluated by 24-1 fractional factorial design. Central composite design (CCD) was used to determine the optimal synthesis conditions, using two different sources of SbV and the significant variables selected in a fractional factorial design. Response surface methodology obtained by CCD provided a model with non-significant regression (p = 0.05) for the synthetic route via KSb(OH)6. On the other hand, synthetic route via SbCl5 reached minimum value of SbIII content of 0.172% and significant regression, and it was selected for further evaluations. The analysis of MA formulations synthesized with SbCl5 under optimized conditions revealed the efficiency of multivariate optimization to reduce SbIII content. In addition, the monitoring of some physicochemical parameters of these formulations maintained at 40 ºC for 90 days, showed that stability was not altered at 95% confidence level.

Minor depression and sleep disorders are common problems in modern societies. There are prescription medicines and herbal remedies to ease the symptoms. St. John-s wort is often used to treat mild depression and valerian root is recommended as a sedative for centuries. Nowadays there is an increasing interest for herbal medicines and therefore the herbal products are becoming more and more popular. As dietary supplement retailers selling low quality products -or even don-t contain what-s claimed on the label- quality becomes the major concern. The legislation to conventional medicines applies to traditional herbal medicines too, which makes registered herbal medicines reliable in production and quality. A CTD application dossier which contains quality data in module 3 should be provided while registering a herbal medicine in EU. In this study we developed a stable formulation and validated two separate methods to quantify the herbal extract amounts in solid oral dosage forms which contains St. John-s Wort and valerian dry extract as active ingredients to comply CTD module 3 requirements.

Cosmetic treatments aim at improving skin appearance through vehicles of good sensory properties. Those vehicles are mainly emulsions and gels designed to deliver safe and effective compounds to skin. Creams and serums are widely used to achieve these goals, but recently a new type of formulation known as organogels triggered scientific attention, particularly in the design of both topical and cosmetic formulations. It has been established that the lipophilic nature of organogels makes it an excellent candidate for the delivery of cosmetic molecules through skin. In this review, we discuss the properties and characteristics of organogels and present the advantages of the application of these systems in cosmetics.Les traitements cosmétiques visent à améliorer l'apparence de la peau grâce à des véhicules dotés de bonnes propriétés sensorielles. Ces véhicules sont principalement des émulsions et gels conçus pour livrer des composants sûrs et efficaces à la peau. Crèmes et sérums sont largement utilisés pour atteindre ces objectifs mais un nouveau type de formulation appelé organogels a récemment attiré l'attention des scientifiques, en particulier en ce qui concerne la conception de formulations à la fois topiques et cosmétiques. Il a été établi que la nature lipophile des organogels en fait d'excellents candidats pour la livraison de molécules cosmétiques à la peau. Dans cette analyse, nous discutons des propriétés et des caractéristiques des organogels, et présentons les avantages de l'utilisation de ces systèmes dans la cosmétique.

Diabetic wounds do not heal in an organized and timely manner due to poor blood glucose control and weakened immune system. In the present study, wound healing properties of curcumin were explored by designing and evaluating a biomimetic composite graft for skin regeneration in diabetic wounds. Curcumin was incorporated in gelatin solution by nanoprecipitation method in particle size of 311.3 ± 35.1 nm. Cytomodulin coupled porous PLGA microparticles (cPMS) were prepared, dispersed in curcumin loaded gelatin solution and cross-linked with glutaraldehyde (GTA) to form a composite graft (Gel-Cur-cPMS). Under SEM, cPMS showed porous structure and Gel-Cur(1%w/v)-cPMS(1%w/v) indicated interconnecting pores of gelatin hydrogel. Formulation stability at the application site was indicated by micro shrinkage temperature. Gel-Cur(1%w/v)-cPMS(1%w/v) absorbed 40% of additional fluid weight of its own weight and demonstrated a sustained release of the curcumin over a period of 7 days. Evaluation of wound healing properties of the composite graft in diabetic wound model indicated 75%, 68% and 63% of wound closure with Gel-Cur(1%w/v)-cPMS(1%w/v), Gel-Cur(1%w/v) and untreated control group respectively. Gel-Cur(1%w/v)-cPMS(1%w/v) demonstrated better and faster wound healing visually along with regenerating a skin of higher tensile strength than Gel-Cur(1%w/v) treated and untreated control group. These results demonstrate that the curcumin-loaded composite graft bears potential as a skin graft with wound healing properties in diabetic wounds.

Liquid protein-based biopharmaceutical formulations have been reported to form aggregation and protein sub-visible particles (SbVPs) during dropping (Randolph et al., J Pharm Sci 2015, 104, 602). However, effects of secondary package on liquid biopharmaceutical formulation stability during dropping are overlooked and have not been reported so far. This study reports the first real-world evaluation on effects of secondary package on liquid biopharmaceutical formulation stability during dropping, using two monoclonal antibodies (mAb-1 and mAb-2) and one fusion protein (FP-1) as model biopharmaceuticals. The potential protective effects of secondary package and formulation composition on liquid biopharmaceutical formulations during dropping were evaluated with micro-flow imaging (MFI) and dynamic light scattering (DLS). The dropping-induced degradation could be detected with the two sensitive particle analyzing techniques MFI and DLS. Formulation compositions have dramatic impact on biopharmaceutical stability during dropping. Surprisingly, unlike the primary packages that have been reported to impact liquid biopharmaceutical stability, the secondary packaging system as described in our current preliminary design has little or no protective effect during dropping. Our study is the first real-world data showing that the secondary package system has little to no effect on the liquid biopharmaceutical formulation quality during dropping. On the contrary, the stability of liquid biopharmaceutical formulations during dropping is more relevant to formulation compositions and primary packages.

Outbreaks of Ebola ebolavirus (EBOV) have been associated with high morbidity and mortality. Milestones have been reached recently in the management of EBOV disease (EVD) with licensure of an EBOV vaccine and two monoclonal antibody therapies. However, neither vaccines nor therapies are available for other disease-causing filoviruses. In preparation for such outbreaks, and for more facile and cost-effective management of EVD, we seek a cocktail containing orally available and room temperature stable drugs with strong activity against multiple filoviruses. We previously showed that (bepridil + sertraline) and (sertraline + toremifene) synergistically suppress EBOV in cell cultures. Here, we describe steps towards testing these combinations in a mouse model of EVD. We identified a vehicle suitable for oral delivery of the component drugs and determined that, thus formulated the drugs are equally active against EBOV as preparations in DMSO, and they maintain activity upon storage in solution for up to seven days. Pharmacokinetic (PK) studies indicated that the drugs in the oral delivery vehicle are well tolerated in mice at the highest doses tested. Collectively the data support advancement of these combinations to tests for synergy in a mouse model of EVD. Moreover, mathematical modeling based on human oral PK projects that the combinations would be more active in humans than their component single drugs.

Previously, a liposomal formulation of a chemotherapeutic agent melphalan (Mlph) incorporated in a fluid lipid bilayer of natural phospholipids in the form of dioleoylglyceride ester (MlphDG) was developed and the antitumor effect was confirmed in mouse models. The formulation composed of egg phosphatidylcholine (ePC), soybean phosphatidylinositol (PI), and MlphDG (8:1:1, by mol) showed stability in human serum for at least 4–5 h. On the contrary, replacing PI with pegylation of the liposomes, promoted fast dissociation of the components from the bilayer. In this work, interactions of MlphDG-liposomes with the most abundant plasma protein—albumin—in function of the presence of PI in the formulation were explored using Fourier transform infrared spectroscopy. The release of MlphDG from the liposomes was studied by asymmetrical flow field-flow fractionation (AF4) using micelles formed by a polyethylene glycol conjugate with phosphatidylethanolamine to mimic the physiological lipid sink like lipoproteins. Our results show that PI actually protects the membrane of MlphDG-liposomes from the protein penetration, presumably due to pairing between the positively charged MlphDG and negatively charged PI, which compensates for the heterogeneity of the lipid bilayer. The AF4 technique also evidences high stability of the formulation as a drug carrier.

CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior O antimoniato de meglumina (AM) é um fármaco de antimônio pentavalente (SbV) recomendado para o tratamento da leishmaniose. Sabe-se que o antimônio trivalente (SbIII), presente como resíduo no AM, contribui para os efeitos colaterais e para o desenvolvimento de resistência a antimoniais. Neste trabalho, planejamento de experimentos (DOE) foi utilizado na síntese de AM para a obtenção de um fármaco com baixos níveis de SbIII residual. Quatro variáveis (fonte de SbV, temperatura, volume de água e pH) e suas interações foram avaliadas preliminarmente através de um planejamento fatorial fracionário 24-1. O planejamento composto central (CCD) foi empregado para determinar as condições ideais de duas rotas sintéticas, usando fontes diferentes de SbV. A análise de formulações de AM sintetizadas em condições otimizadas, revelou a eficiência do DOE para reduzir os níveis de SbIII residual. Além disso, o monitoramento de alguns parâmetros físico-químicos, i.e., teor de SbIII, pH e osmolaridade dessas formulações, mantidas a 40 oC por 90 dias, mostrou com 95% de confiança que a estabilidade não foi alterada. Teor de SbIII, em relação à quantidade de Sb total, presente nos compostos de AM foi usado como resposta no DOE e na avaliação da estabilidade de formulações de AM. Para esse fim, foi desenvolvido e validado um procedimento para especiação de Sb inorgânico em AM empregando espectrometria de absorção atômica por geração de hidretos (HG-AAS). A especiação do Sb ocorre em duas etapas: 1) quantificação de Sb total usando um agente redutor (KI); 2) quantificação de SbIII pela geração seletiva de estibina (SbH3) com o uso de ácido cítrico, em meio com elevada concentração de SbV. A concentração de ácido cítrico utilizada no método otimizado foi de 4 a 20 vezes menor do que a recomendada na literatura, evitando o entupimento do capilar da célula de quartzo T. Os limites de quantificação (LQ) foram calculados em 0,48 μg L-1 para Sb total e 0,17 μg L-1 para SbIII. Os valores de desvio padrão relativo (DPR) variaram de 3,1 a 19,6% e 9,1 a 20,1%, enquanto a recuperação variou de 95,6 a 102,3% e 89,1 a 108,1%, para Sb total e SbIII, respectivamente. Meglumine antimoniate (MA) is a pentavalent antimony (SbV) drug recommended for the treatment of leishmaniasis. It is known that the trivalent antimony (SbIII) present as a residue in MA contributes to the drug side effects and to the development of antimonial drug resistance. In this work, design of experiments (DOE) was used in the synthesis of MA in order to obtain a drug with low levels of SbIII. Four variables (source of SbV, volume, temperature, water volume and pH) and their interactions were preliminarily evaluated by 24-1 fractional factorial design. Central composite design (CCD) was used to determine the optimal conditions of two synthetic routes, using different sources of SbV. The analysis of MA formulations synthesized under optimized conditions revealed the efficiency of DOE to reduce SbIII content. In addition, the monitoring of some physicochemical parameters, i.e., SbIII content, pH and osmolarity of these formulations maintained at 40 oC for 90 days, showed that stability was not altered at 95% confidence. The SbIII content, in relation to the amount of total Sb, present in MA compounds was used as a response in the DOE and in the evaluation of the MA formulations stability. For this purposte, a procedure for speciation of inorganic Sb in MA was developed and validated using hydride generation atomic absorption spectrometry (HG-AAS). The Sb speciation occurs in two stages: 1) quantification of total Sb using a reducing agent (KI); 2) quantification of SbIII by the selective generation of stibin (SbH3) with the use of citric acid, in medium with higher SbV concentration. The concentration of citric acid used in the optimized method was 4 to 20-fold lower than that recommended in the literature, preventing clogging of the capillary of the T quartz cell. The limits of quantification (LOQ) were calculated at 0.48 μg L-1 for total Sb and 0.17 μg L-1 for SbIII. The relative standard deviation (RSD) values ranged from 3.1 to 19.6% and 9.1 to 20.1%, while recovery ranged from 95.6 to 102.3% and 89.1 to 108.1%, for total Sb and SbIII, respectively.

Nanoparticles (NPs) offer noteworthy advantages in the treatment of several diseases by prompting, among other benefits, the site-specific delivery of drugs. Ultra-small nanostructured lipid carriers (usNLCs) are no exception. These correspond to a class of NPs composed of a blend of solid and liquid lipids, the latter usually in a higher proportion, which promotes a less ordered solid lipid matrix, providing a higher drug loading capacity, drug release modulation, and improved stability in comparison with other lipid nanoparticles. Several manufacturing methods have been described for obtaining usNLCs. However, a comprehensive understanding of the process is imperative to warrant the final quality of the NPs. In the present work, the hot high pressure homogenization (HPH) method, which is characterized by easy scaling-up, simplicity and ease of use, was used for the development of highly concentrated small size NLCs. Critical process parameters (CPPs) and critical material attributes (CMAs) were evaluated to investigate the manufacturing process reproducibility, inter-batch consistency, long-term formulation stability, drug loading capacity and drug release. To gain a broader understanding of this method, multivariate analysis was applied to investigate how the physicochemical properties of the usNLC are affected by the variation in CPPs/CMAs. CPPs include HPH-time, and HPH-pressure, while CMAs, such as lipid content, are also taken into consideration. The results show that a high lipid content (15% w/w), an intermediate pressure and a short HPH time seem to be the crucial parameters to obtain both a small particle size ( |30| mV).

Small angle neutron scattering (SANS) studies of a model pharmaceutical formulation reveal how formulation stability depends on the compatibility of individual components. Solutions of two common protein formulation excipients, polysorbate 80 (PS80), a nonionic surfactant that prevents aggregation, and m-cresol, an antimicrobial agent for multi-dose injectable formulations, are investigated. The addition of m-cresol to PS80 solutions leads to solution turbidity and irreversibly alters PS80 micelle morphology. This slow preservative-induced destabilization of PS80 micelles progresses over days or even weeks, which highlights the essential role that aggregation kinetics plays in preservative-surfactant interactions. The temperature-dependence of PS80 micelle growth kinetics is quantified by SANS in the presence of m-cresol. Aggregation is a two-step process, where initial formation of small aggregates is followed by a period of monotonic power-law growth, providing evidence for the mechanism. Total aggregate mass stays constant after initial aggregate formation, and addition of a pH-regulating citrate buffer dramatically accelerates aggregation kinetics.

We describe the preparation of injectable polymeric paste (IPP) formulations for local and sustained release of drugs. Furthermore, we include the characterization and possible applications of such pastes. Particular attention is paid to characteristics relevant to the successful clinical formulation development, such as viscosity, injectability, degradation, drug release, sterilization, stability performance and pharmacokinetics. Paste injectability was characterized using measured viscosity and the Hagen-Poiseuille equation to determine injection forces. Drug degradation, release and formulation stability experiments were performed in vitro and drug levels were quantified using HPLC-UV methods. Pharmacokinetic evaluation of sustained-release lidocaine IPPs used five groups of six rats receiving increasing doses subcutaneously. An anti-cancer formulation was evaluated in a subcutaneous tumor xenograft mouse model. The viscosity and injectability of IPPs could be controlled by changing the polymeric composition. IPPs demonstrated good long-term stability and tunable drug-release with low systemic exposure in vivo in rats. Preliminary data in a subcutaneous tumor model points to a sustained anticancer effect. These IPPs are tunable platforms for local and sustained delivery of drugs and have potential for further clinical development to treat a number of diseases.

The compatibility of formulation components is crucial for safe and high-quality medicines. To detect the potential for incompatibility and to assess formulation stability, it is beneficial to conduct a compatibility study during the drug development phase. The therapy of tuberculosis normally consists of two or more medicines taken together. Consequently, different antituberculotic fixed-dose combination (FDC) formulations have been developed. Isoniazid is first-line medicine and present in several FDCs. Low bioavailability due to the active substances’ incompatibility in acidic medium was reported for some of these FDC forms. Rifabutin, also a first- line antituberculotic, is available in the market as a single component formulation. This study presents compatibility testing of these two active substances for a new FDC and evaluates the impact of the most common solid dosage forms’ excipients on the stability of two active substances. The potential for interaction between the formulation components was analyzed by the UHPLC method. One degradation product and one interaction product were observed and further characterized by high- resolution mass spectrometry. Still, significant degradation of two active substances, such as reported in marketed FDC formulations was not detected for this combination. The stability and drug delivery of the proposed combination were confirmed by the dissolution test in acidic medium.

One of the major challenges in formulation development of biopharmaceuticals is improving long-term storage stability, which is often achieved by addition of excipients to the final formulation. Finding the optimal excipient for a given protein is usually done using a trial-and-error approach, due to the lack of general understanding of how excipients work for a particular protein. Previously, preferential interactions (binding or exclusion) of excipients with proteins were postulated as a mechanism explaining diversity in the stabilisation effects. Weak preferential binding is however difficult to quantify experimentally, and the question remains whether the formulation process should seek excipients which preferentially bind with proteins, or not. Here, we apply solution NMR spectroscopy to comprehensively evaluate protein-excipient interactions between therapeutically relevant proteins and commonly used excipients. Additionally, we evaluate the effect of excipients on thermal and colloidal protein stability, on aggregation kinetics and protein storage stability at elevated temperatures. We show that there is a weak negative correlation between the strength of protein-excipient interactions and effect on enhancing protein thermal stability. We found that the overall protein-excipient binding per se can be a poor criterion for choosing excipients enhancing formulation stability. Experiments on diverse set of excipients and test proteins reveal that while excipients affect all of the different aspects of protein stability, the effects are very much protein specific, and care must be taken to avoid apparent generalizations if a smaller dataset is being used.

The formulation, physicochemical stability and bioaccessibility of astaxanthin (AST) loaded oil-in-water nanoemulsions fabricated using gypenosides (GPs) as natural emulsifiers was investigated and compared with a synthetic emulsifier (Tween 20) that is commonly applied in food industry. GPs were capable of producing nanoemulsions with a small volume mean diameter (d4,3 = 125 ± 2 nm), which was similar to those prepared using Tween 20 (d4,3 = 145 ± 6 nm) under the same high-pressure homogenization conditions. GPs-stabilized nanoemulsions were stable against droplet growth over a range of pH (6–8) and thermal treatments (60–120 °C). Conversely, instability occurred under acidic (pH 3–5) and high ionic strength (25–100 mM CaCl2) conditions. In comparison with Tween 20, GPs were more effective at inhibiting AST from degradation during 30 days of storage at both 5 and 25 °C. However, GPs led to lower lipid digestion and AST bioaccessibility from nanoemulsions than did Tween 20.

Biopharmaceuticals are one of the fastest growing areas within the pharmaceutical industry. As protein drugs require parenteral administration, they are commonly formulated as aqueous solutions. However, this is not always feasible due to their general instability. In such cases, lyophilised powders for injection are the dosage form of choice, for the preparation of stable products. Lyophilisation is known to be highly time and energy consuming, and hence it is an expensive technological process. Thus, the pharmaceutical industry is increasingly focused on its optimisation. Implementation of aggressive conditions, together with optimisation of formulation parameters, represent the contemporary approach to reduction of the primary drying time. As such, incorporation of drug-specific excipients can contribute significantly to the stability of a biologically active ingredient, and indirectly they can also affect the time needed for lyophilisation. The addition of the most relevant protein stabilisers, surfactants, buffers and bulking agents is therefore crucial. The main aim of the present review is to define the most important groups of biopharmaceutical excipients, based on their roles in formulations and the mechanism(s) through which they support the lyophilisation process, to provide products with the required protein efficiency and product characteristics. The scope of the article is to critically discuss the suitability of novel stabilizers, with higher critical temperatures and bulking agents in terms of implementation of aggressive primary drying. For better assignment of the topic-related challenges, the stabilities of biopharmaceutical drugs and the fundamentals of the lyophilisation process are also briefly described.

In this work, we compared the formulation, stability and bioaccessibility of ergocalciferol nanodispersions using modified lecithin (ML) and sodium caseinate (SC) as natural emulsifiers. The mean particle size of nanodispersions stabilized by ML (56 nm) was much smaller than those stabilized by SC (112 nm). The ML-stabilized nanodispersions were stable over a wide range of pH, NaCl concentrations and heating, but became unstable with slight increase in particle size when exposed to CaCl 2 solution. In comparison, SC-stabilized nanodispersions were relatively unstable to particles aggregation at pH 4 and 5, CaCl 2 addition and heating. Long-term stability for ergocalciferol were observed in both ML- and SC-stabilized nanodispersions. In the absence of milk, the ergocalciferol bioaccessibility was strongly dependent on the emulsifier type, with ML providing much higher bioaccessibility than SC. During in vitro gastrointestinal digestion, the incorporation of milk into nanodispersions could increase the bioaccessibility and stability for ergocalciferol.

Space agencies are working intensely to push the current boundaries of human spaceflight by sending astronauts deeper into space than ever before, including missions to Mars and asteroids. Spaceflight alters human physiology due to fluid shifts, muscle and bone loss, immune system dysregulation, and changes in the gastrointestinal tract and metabolic enzymes. These alterations may change the pharmacokinetics and/or pharmacodynamics of medications used by astronauts and subsequently might impact drug efficacy and safety. Most commonly, medications are administered during space missions to treat sleep disturbances, allergies, space motion sickness, pain, and sinus congestion. These medications are administered under the assumption that they act in a similar way as on Earth, an assumption that has not been investigated systematically yet. Few inflight pharmacokinetic data have been published, and pharmacodynamic and pharmacokinetic/pharmacodynamic studies during spaceflight are also lacking. Therefore, bed-rest models are often used to simulate physiological changes observed during microgravity. In addition to pharmacokinetic/pharmacodynamic changes, decreased drug and formulation stability in space could also influence efficacy and safety of medications. These alterations along with physiological changes and their resulting pharmacokinetic and pharmacodynamic effects must to be considered to determine their ultimate impact on medication efficacy and safety during spaceflight.

A new institutional clinical trial assessed the improvement of sleep disorders in 40 children with autism treated by immediate-release melatonin formulation in different regimens (0.5 mg, 2 mg, and 6 mg daily) for one month. The objectives of present study were to (i) prepare low-dose melatonin hard capsules for pediatric use controlled by two complementary methods and (ii) carry out a stability study in order to determine a use-by-date. Validation of preparation process was claimed as ascertained by mass uniformity of hard capsules. Multicomponent analysis by attenuated total reflectance Fourier transformed infrared (ATR-FTIR) of melatonin/microcrystalline cellulose mixture allowed to identify and quantify relative content of active pharmaceutical ingredients and excipients. Absolute melatonin content analysis by high performance liquid chromatography in 0.5 mg and 6 mg melatonin capsules was 93.6%±4.1% and 98.7%±6.9% of theoretical value, respectively. Forced degradation study showed a good separation of melatonin and its degradation products. The capability of the method was 15, confirming a risk of false negative, Graphical abstract fx1

High protein titers are gaining importance in biopharmaceutical industry. A major challenge in the development of highly concentrated mAb solutions is their long-term stability and often incalculable viscosity. The complexity of the molecule itself, as well as the various molecular interactions, make it difficult to describe their solution behavior. To study the formulation stability, long- and short-range interactions and the formation of complex network structures have to be taken into account. For a better understanding of highly concentrated solutions, we combined established and novel analytical tools to characterize the effect of solution properties on the stability of highly concentrated mAb formulations. In this study, monoclonal antibody solutions in a concentration range of 50–200 mg/ml at pH 5–9 with and without glycine, PEG4000, and Na2SO4 were analyzed. To determine the monomer content, analytical size-exclusion chromatography runs were performed. ζ-potential measurements were conducted to analyze the electrophoretic properties in different solutions. The melting and aggregation temperatures were determined with the help of fluorescence and static light scattering measurements. Additionally, rheological measurements were conducted to study the solution viscosity and viscoelastic behavior of the mAb solutions. The so-determined analytical parameters were scored and merged in an analytical toolbox. The resulting scoring was then successfully correlated with long-term storage (40 d of incubation) experiments. Our results indicate that the sensitivity of complex rheological measurements, in combination with the applied techniques, allows reliable statements to be made with respect to the effect of solution properties, such as protein concentration, ionic strength, and pH shift, on the strength of protein-protein interaction and solution colloidal stability.

In this paper, microencapsulated formulation of Bacillus thuringiensis subsp. kurstaki were prepared by the emulsion gelling method. Stability of formulation under ultraviolet (UV) radiation by spore viability and mortality teston Ephestia kuehniella Zeller larvae are investigated. Response surface methodology was used for optimising the formulations. Calcium Chloride (0.1 and 0.3 M) and sodium alginate (2 and 3% w/w) were used in the formulations, and stirring speed was selected between 1500 and 2000 rpm. Morphology of the microcapsule was evaluated by light microscopy. The optimal values for the variables were: sodium alginate = 2.15%, CaCl2 = 0.24 M, speed = 1606 rpm and spore viability, mortality and diameter of 68% ± 1.73, 85% ± 1.5 and 9μ ± 1.3, respectively; while the predicted values by the software were 70% for spore viability, 87% for mortality and 8.3 μ for diameter.

The mucosal surfaces represent the major site of entry of many pathogens, and major challenges in vaccine development include safety and stability in a suitable dosage form. Micro- and nanocarrier-based delivery systems as nasal vaccines induce humoral, cellular, and mucosal immunity. The nasal route of vaccination could also offer immunity at several distant mucosal sites (oral, rectal, vaginal, and pulmonary), which is considered a simplified and cost-effective mode of vaccination with enhanced patient compliance. Most of the nasal vaccine delivery systems in the form of microparticulates, nanoparticulates, and liposomes are currently under development and prove to offer immunity in animal models. The importance and potential of the nasal route of administration for vaccines is unexplored, and this chapter outlines the opportunities, challenges, and potential delivery solutions to facilitate the development of improved nasal vaccines for infectious diseases.