The increasing prohibition of animal testing for cosmetic products has driven the development of alternative approaches to ensure consumer safety. Skin sensitization is one of the most critical toxicological endpoints to evaluate, requiring rigorous assessment to ensure the safety of cosmetic ingredients. This study proposes an Integrated Testing Strategy (ITS) that combines in chemico (Direct Peptide Reactivity Assay—DPRA) and in silico approaches (a six-platform computational panel) to evaluate the sensitization potential of substances. Initially, the in chemico methodology was validated through a partial proficiency demonstration to ensure experimental reliability. Subsequently, this ITS was applied to the Baccharis trimera extract and its major marker, 3-Caffeoylquinic acid (chlorogenic acid/3-CQA). Our results demonstrate that the DPRA alone is insufficient to classify the sensitization potential of complex mixtures, as recommended by OECD guidelines. The integration of in silico data proved essential to interpret the reactivity of the botanical matrix, revealing that the sensitization potential observed in the extract does not stem solely from 3-CQA, but likely results from the synergistic contribution of more lipophilic caffeoylquinic acid isomers. This approach demonstrates that integrating experimental and computational methods is fundamental for a robust safety assessment, offering an efficient, animal-free strategy for the early screening of cosmetic ingredients and for refining the interpretation of toxicological data in complex chemical environments.

In vitro release testing (IVRT) serves as a crucial tool to assess the quality, physicochemical behavior, and performance of semisolid formulations already available on the market. In vitro skin permeation studies (IVPT) are widely used to evaluate the safety and efficacy profiles of topical drugs, utilizing biological membranes prepared from ex vivo human and porcine skin tissues. This study aimed to develop and validate a discriminative IVRT method to evaluate various marketed topical benzoyl peroxide formulations. Additionally, IVPT was employed to assess skin permeation and retention profiles of these formulations, comparing porcine skin results with those obtained by using ex vivo human skin tissues. Physicochemical differences among the evaluated benzoyl peroxide formulations were identified, with the poloxamer-based formulation exhibiting a higher release rate. IVPT using both porcine and human skin differentiated retention and skin permeation profiles, with the poloxamer-based formulation demonstrating greater skin retention capacity compared to the other formulations evaluated. Similar conclusions on benzoyl peroxide retention and cutaneous permeation were drawn from both porcine and human skin IVPT tests, confirming the correlation between the two models.

The present work aims to establish a formulation-specific, physiologically based pharmacokinetic (PBPK) model for efavirenz (EFV) nanocrystals that have shown increased dissolution and were produced following a top-down approach based on wet milling and spray drying by integrating solid-state characterization, in vitro performance, and preclinical pharmacokinetics to enable translational predictions in humans. The resulting material was thoroughly characterized using diffraction-based, spectroscopic, thermal, morphological, and particle sizing techniques, along within vitro dissolution testing and an in vivo pharmacokinetic analysis in rats. Then, a fully rat PBPK model was constructed using GastroPlus and incorporating the biopharmaceutical nanoparticle properties through the product particle size distribution (P-PSD) approach. The physiologically based biopharmaceutics model (PBBM) was validated with rat in vivo data and subsequently extrapolated to simulate human physiology. Compared with unprocessed EFV, nanocrystals exhibited superior dissolution efficiency (90.4% vs 52.6%) and a more homogeneous size distribution. Furthermore, the in vivo studies confirmed an increase in EFV exposure. The rat PBPK model accurately reproduced plasma profiles of both formulations, with all predictive error metrics falling within the acceptable 2-fold range. Extrapolation to human physiology revealed that a 350 mg EFV NC dose achieved systemic exposure comparable to that of standard 600 mg immediate-release tablet, but with faster absorption. Sensitivity analyses highlighted the critical influence of particle size and bile salt solubilization capacity on EFV oral absorption. This study pioneers the application of a fully mechanistic PBPK/PBBM model tailored to nanocrystal formulations of EFV. By bridging preclinical and human data through in silico simulation, the proposed approach supports dose optimization strategies and reinforces the role of nanotechnology in advancing nonbiological complex drug development.

Application of software for toxicity analysis of cyanobacterial toxin remains poorly explored compared to such approach in other areas. Predictive software has been extensively used in the discovery of new drugs and assessment of cosmetics safety. Cosmopolitan microcystin variants have well-defined chemical structures, and their toxicity has been measured through in vivo and in vitro experiments. Toxic effects of unexplored variants are estimated using toxicity values from MC-LR, currently recognized as the most toxic and extensively studied microcystin. In silico software facilitates the acquisition of toxicity information for molecules with elucidated chemical structures expeditiously, reproducibly, and at reduced cost, thereby reducing the number of animals required for analyses. The objective of this study was to compare six prediction simulators to determine which ones generate useful information regarding microcystin toxicity. Two-dimensional chemical structures of four microcystin variants (MC-LR, MC-RR, MC-YR, and MC-HarHar) were used in selected software (ADMET Predictor®, ADMETlab, admetSAR, SwissADME, T.E.S.T., and ECOSAR) to obtain predictions on microcystin parameters. After assessing the applicability domain of each software, ADMET Predictor®, admetSAR, SwissADME, and T.E.S.T. were found adequate for predicting lipophilicity, permeability, intestinal absorption, transport proteins, and environmental biodegradation. ADMET Predictor®, admetSAR, and T.E.S.T. yielded similar results despite having fewer than 35 available models, and thus were found to be more consistent for microcystins than SwissADME, that showed the most discrepant results. Conversely, ADMET Predictor® demonstrated over 70 models with valid predictions. Our findings indicate that in silico prediction software with valid applicability domains is promising for generating information regarding toxicity differentiation of microcystin variants.

The aim of this study was to use the physiologically based biopharmaceutics modeling (PBBM) strategy to evaluate the biopredictive power of different dissolution test conditions for immediate release tablets containing 20 mg of rivaroxaban. The model was developed and validated with literature data and used to predict plasma concentration-time curves of reference drug product Xarelto® 20 mg based on the experimental dissolution profiles. Virtual bioequivalence studies were run considering each dissolution profile under fasted and fed conditions to evaluate the biopredictive power of each dissolution method. The dissolution profiles and the results of the virtual bioequivalence studies showed that it is necessary to have a minimal amount of surfactant (0.15%), rotation speed of at least 60 rpm and 900 mL of acetate buffer solution pH 4.5 to achieve biopredictive condition. Using the PBBM approach, it was possible to evaluate the differences of the dissolution test conditions, and its influence on drug bioavailability, setting up a dissolution specification for rivaroxaban immediate release tablets.

Orally disintegrating films (ODFs) were prepared from hydroxypropyl methylcellulose (HPMC E6 1.5%, 2.0%, and 2.5%), plasticizers ((glycerin (Gly), propylene glycol (PP), or polyethylene glycol (PEG)), and isosorbide dinitrate using the solvent casting method. Design of experiments (DoE) was used considering the amount of film-forming agent (HPMC) and the nature of the plasticizers as independent variables and thickness, mechanical properties, disintegration time, and dissolution efficiency as dependent variables. The best formulation was selected based on the desirability function (f(D)). Color analysis was performed using CIE-Lab coordinates. DSC curves and XRPD diffractograms showed ISDN amorphization, probably due to complexation with polymers (HPMC E6 and HP beta CD). The polymer (HPMC E6) and plasticizer played a critical role in the mechanical and optical properties of the films; however, these factors had no significant effect on the dissolution efficiency of ISDN. Principal component analysis revealed a more defined distinctionof the ODFs according to their chromatic characteristics. ODFs prepared with Gly and PEG 400 were translucent, whereas the other films were transparent. The dissolution efficiency values of the ODFs (F1-9) were higher than those observed for the reference product (88.35.0.01%), indicating increased solubility of ISDN, probably due to complexation with polymers (HPMC E6 and HP beta CD). The maximum value obtained for desirability function (f(D))) was 0.86, which corresponded to the use of 1.5% to 2.0% polymer (HPMC E6) and Gly or PP as plasticizers. The F5 formulation showed adequate thickness (85.00 +/- 6.38.m), elongation at break (4.82 +/- 0.01%), adhesiveness (0.23 +/- 0.09 mJ), dissolution efficiency (95.97 +/- 0.01%), and high transparency.

In this study, orodispersible films formed from hydroxypropyl methylcellulose (HPMC) E6 (2, 2.5, and 3%) and plasticizers ((glycerin (Gly), propylene glycol (PP), or polyethylene glycol (PEG)), containing doxazosin mesylate, were prepared by the solvent casting method and characterized. Design of experiments (DoE) was used as a statistical tool to facilitate the interpretation of the experimental data and allow the identification of optimal levels of factors for maximum formulation performance. Differential scanning calorimetry (DSC) curves and X-ray powder diffraction (XRPD) diffractograms showed doxazosin mesylate amorphization, probably due to complexation with the polymer (HPMC E6), and the glass transition temperature of the polymer was reduced by adding a plasticizer. Fourier transformed infrared (FTIR) spectroscopy results showed that the chemical structure of doxazosin mesylate was preserved when introduced into the polymer matrix, and the plasticizers, glycerin and PEG, affected the polymer matrix with high intensity. The addition of plasticizers increased the elongation at break and adhesiveness (Gly > PEG > PP), confirming the greater plasticizer effect of Gly observed in DSC and FTIR studies. Greater transparency was observed for the orodispersible films prepared using PP. The addition of citric acid as a pH modifier was fundamental for the release of doxazosin mesylate, and the desirability formulation had a release profile similar to that of the reference product.

Hydrochlorothiazide (HTZ) and Valsartan (VAL) are poorly soluble drugs in BCS classes IV and II. This study aimed to develop a method to assess the dissolution profile of tablets containing HTZ (12.5 mg) and VAL (160 mg) as a fixed-dose combination, using in silico tools to evaluate products marketed in Brazil and Peru. Firstly, in vitro dissolution tests were performed using a fractional factorial design 33−1. Then, DDDPlus™ was used to carry out experimental design assays of a complete factorial design 33. Data from the first stage were used to obtain calibration constants for in silico simulations. The factors used in both designs were formulation, sinker use, and rotation speed. Finally, effects and factor interaction assessment was evaluated based on a statistical analysis of the dissolution efficiency (DE) obtained from simulations. Thus, the established final conditions of the dissolution method were 900 mL of phosphate buffer pH 6.8, 75 rpm of rotation speed, and sinker use to prevent formulation floating. The reference product stood out because of its higher DE than other formulations. It was concluded that the proposed method, in addition to ensuring total HTZ and VAL release from formulations, has adequate discriminative power.

This study aimed to develop a biopredictive dissolution method for desvenlafaxine ER tablets using design of experiments (DoE) and physiologically based biopharmaceutics modeling (PBBM) to address the challenge of developing generic drug products by reducing the risk of product failure in pivotal bioequivalence studies. For this purpose, a PBBM was developed in GastroPlus® and combined with a Taguchi L9 design, to evaluate the impact of different drug products (Reference, Generic #1 and Generic #2) and dissolution test conditions on desvenlafaxine release. The influence of the superficial area/volume ratio (SA/V) of the tablets was observed, mainly for Generic #1, which presented higher SA/V than the others, and a high amount of drug dissolved under similar test conditions. The dissolution test conditions of 900 mL of 0.9% NaCl and paddle at 50 rpm with sinker showed to be biopredictive, as it was possible to demonstrate virtual bioequivalence for all products, despite their release-pattern differences, including Generic #3 as an external validation. This approach led to a rational development of a biopredictive dissolution method for desvenlafaxine ER tablets, providing knowledge that may help the process of drug product and dissolution method development.

The use of physiologically based biopharmaceutics modeling (PBBM) and bioequivalence safe space is increasingly common for immediate-release drug products. However, for extended-release (ER) formulations there are only a few examples of this application. In this study, we developed ER formulations containing cyclobenzaprine 15 mg, supported by PBBM and bioequivalence safe space. Four formulations were prepared, F1, F2, F3 (ER mini-tablet formulations) and F4 (ER tablet formulation), and the dissolution profiles were evaluated. The dissolution profile of the reference drug product was also evaluated and used to set a bioequivalence safe space. A PBBM was set up, evaluated, and used to predict the in vivo behavior of the formulations. The bioequivalence safe space was calculated to be between - 25% and + 75% of the k1 and Tlag values of the dissolution profile of the reference drug product when applying the first-order dissolution kinetic model. All time points of the dissolution profile of the ER mini-tablet formulation F2, were within the safe space, and was approved in 10 of 10 trials of crossover virtual bioequivalence studies. Based on the PBBM strategy and bioequivalence safe space, it was possible to develop an ER mini-tablet formulation virtually bioequivalent to the reference drug product, even though this formulation failed the f2 test.

Oseltamivir phosphate is used to treat influenza. For registration of a generic product, bioequivalence studies are crucial, however, in vitro studies can sometimes replace the conventional human pharmacokinetic. To assess whether the dissolution profile is comparable with the in vivo release, physiologically based pharmacokinetic absorption models (PBPK) are being used. The aim of the study was to develop a generic capsule of oseltamivir phosphate 30 mg with process understanding and control, development of PBPK model and comparison of virtual bioequivalence study (VBE) to the real bioequivalence study that was also performed. For that, 30 mg capsules were prepared by wet granulation according to 2(2) full factorial design. The biobatch was prepared with the selected process and a batch was made with the API from the second manufacture. Both manufactures presented polymorph A and the second manufacture showed higher particle size. Product batches produced without adding water during granulation showed higher dissolution. The addition of water associated with higher conical mill speed, lowered the average weight of the capsules. The biobatch dissolution was similar to Tamiflu; also, they were bioequivalent. The crossover VBE between the biobatch and Tamiflu corroborated with the real bioequivalence study. The same result was found for the batch with higher particle size. PBPK model showed that computer simulations can help pharmaceutical companies to replace in vivo studies.

In this study, we investigated the influence of the olfactive stimulus on visual attention. Two groups of 30 subjects participated in two experiments. Both experiments presented two arrays of fruits stimulus intercalated by an olfactive intervention. The stimulus was received in the form of images by the first group and in the form of words by the second group. An eye-tracking device monitored the timekeeping of visual attention dispensed in each stimulus. The results showed that olfactive priming influenced visual attention in both cases but with a greater degree in the images stimulus group. This study shows for the first time that image information is more susceptible to priming olfactive information than wording information. This effect may be associated with the formation of mental images in working memory, aroused by fragrances.

The development of extended-release dosage forms with adequate drug release is a challenge for pharmaceutical companies, mainly when the drug presents high solubility, as in Biopharmaceutics Classification System (BCS) class I. This study aimed to develop extended-release mini-tablets containing metoprolol succinate (MS), while integrating design of experiments (DOE) and physiologically based biopharmaceutics modeling (PBBM), to predict its absorption and to run virtual bioequivalence (VBE) studies in both fasted and fed states. Core mini-tablet formulations (F1, F2, and F3) were prepared by direct compression and coated using nine coating formulations planned using DOE, while varying the percentages of the controlled-release and the pore-forming polymers. The coated mini-tablets were submitted to a dissolution test; additional formulations were prepared that were optimized by simulating the dissolution profiles, and the best one was submitted to VBE studies using GastroPlus® software. An optimized formulation (FO) containing a mixture of immediate and extended-release mini-tablets showed to be bioequivalent to the reference drug product containing MS when running VBE studies in both fasted and fed states. The integration of DOE and PBBM showed to be an interesting approach in the development of extended-release mini-tablet formulation containing MS, and can be used to rationalize the development of dosage forms.

Objective. - Our objective was to evaluate the absorption of finasteride administered by oral and topical routes for treatment of androgenetic alopecia, by means of computer simulations using the GastroPlus (R) software. Material and methods. - In vivo plasma concentration profile of oral administration of finasteride 1 mg tablets from the literature was used in the software to build a compartmental pharmacokinetic model that was extrapolated to simulate topical administration of finasteride 0.25% solution in the scalp. Results were compared to literature and other drug concentrations (0.1% and 1%) were also predicted. Results. - Compared to literature data, predictive plasma curve from oral administration of finasteride 1 mg showed good correlation, (R2)=0.992, and Cmax=4.6769 ng/mL. Simulations of topical administration in the scalp for finasteride 0.25% solution showed good correlation, R-2=0.908, and Cmax=0.04325 ng/mL when compared to literature data. Topical administration was also simulated at concentrations 0.1% and 1%, both with low plasma concentrations. Conclusion. - The results obtained in this study suggest that topical finasteride is a potential treatment for androgenetic alopecia in the concentrations analyzed using computer simulations in GastroPlus (R). (c) 2021 Academie Nationale de Pharmacie. Published by Elsevier Masson SAS. All rights reserved.

Objective This work aims to evaluate the ability of biorelevant dissolution media to simulate the bioavailability of efavirenz tablets, establish an in vitro-in vivo relationship (IVIVR) based on in vivo data using GastroPlus(R) and simulate formulation changes using DDDPlus (TM). Methods Solubility and drug release profiles were conducted in SLS 0.5% and biorelevant media, such as FaSSIF, FeSSIF, FaSSIF-V2, and FeSSIF-V2. The efavirenz physicochemical properties were used to simulate the plasma concentration profile and compare the simulated pharmacokinetic parameters in fasted and fed states. An IVIVR was developed using Loo-Riegelman as the deconvolution method to estimate drug bioavailability. DDDPlus (TM) was used to perform virtual trials of formulations to evaluate whether formulations changes and the efavirenz particle size could influence the bioavailability. Results The drug dissolution displayed higher levels in the biorelevant media that simulated gut-fed state (FeSSIF and FeSSIF-V2). The absorption model successfully predicted the efavirenz pharmacokinetics, and FeSSIF-V2 was chosen as the predictive dissolution media, while an IVIVR was established using the Loo-Riegelman deconvolution method. Conclusions The present work provides valuable information about efavirenz solubility and kinetics in the gastrointestinal tract, allowing an IVIVR to support future formulation changes. This understanding is essential for rational science-driven formulation development. At least, this study also showed the validity and applicability of in vitro and in silico tools in the regulatory scenario helping on drug development.

A range of liquid emollient ester-loaded body emulsions were evaluated on the basis of sensory profile, consumer perception, physicochemical, biometrological, and rheological performances. This was complemented by skin hydration and transepidermal water loss (TEWL) studies performed using a Corneometer (R) and Tewameter (R), respectively. The rheological studies showed similar behavior of creams on variation of shear stress, frequency, and temperature. However, skin hydration performances of creams were found varied when compared between base cream and emollient ester-containing creams. In case of the TEWL study, base cream (F1, no ester) and octyl palmitate-containing cream (F2), exhibited the most protective behavior for the skin with minimal water loss. Sensory profiles of creams were obtained using the sensory descriptive analysis method. A study performed with human consumers (as subjects) using the difference of test attributes (directional) method showed the importance of sensory evaluation with consumers and proved to be an interesting analysis for comparison with the sensorial profile. In this case, the attributes with the highest intensity in the evaluated creams were, that is, spreadability, slipping, and dry touch. Stability studies of creams were performed for 90 days under different storage conditions. Stability studies showed octyl stearate-containing (F2) and palmitate-containing (F5) creams exhibited best stability under extended storage conditions. This research has demonstrated the importance of physicochemical characterization and sensory evaluations performed with consumers for more targeted analysis of cream-based formulations to differentiate product characteristics and identify attributes-led preferences. Practical applications Results from the present work suggest that emollients are directly responsible for sensory characteristics such as spreading, absorption, and touch on the skin; they can influence the viscosity and appearance of emulsions. After application on the skin, they regulate the moisture content of the epidermis together with the humectants. This research has demonstrated the importance of sensory evaluation, especially performed with consumers, which may be considered a more targeted analysis to differentiate product characteristics and help in the choice of emollient with the desired characteristics for the cosmetic product.

In silico methodologies can be used in the discovery of new drugs for measuring toxicity, predicting effects of substances not yet analyzed by in vivo methodologies. The ADMET Predictor® software (absorption, distribution, metabolism, elimination, and toxicity [ADMET]) was used in this work to predict toxic effects of microcystin variants MC-LR, MC-YR, MC-RR, and MC-HarHar. In the case of rodents, predictive results for all analyzed variants indicated carcinogenic potential. The predictive model of respiratory sensitivity in this group differentiated microcystins into 2 categories: sensitizer (MC-LR and -YR) and non-sensitizer (MC-HarHar and -RR). Predictive results for humans indicated that MC-LR and -RR are phospholipidosis inducers; on the other hand, MC-LR showed the highest predictive value of permeability in rabbit cornea and probability of crossing lipoprotein barriers (MC-LR>-YR>-HarHar>-RR). Considering bioavailable fractions, microcystins are more likely to cause biological effects in rats than humans, showing significant differences between models. The results of ADMET predictions add valuable information on microcystin toxicity, especially in the case of variants not yet studied experimentally.

The treatment of schistosomiasis is based on a single drug, the praziquantel (PZQ), an oral bioavailable and efficient agent which causes minimal side effects. The main concern about this approach, however, is that relying on only one drug to treat a helminthic disease is a dangerous strategy since history shows that pathogens easily evolve to resistant forms. Actually, reports about experimental strains exhibiting low sensibility to PZQ can be found in literature. The search for new antischistosomals, consequently, is urgent. Here we report the synthesis of seventeen Schiff bases of 4-(4-Substituted phenyl)-N-(4-substituted benzylidene)thiazole-2-amines which were tested in vitro and in vivo against Schistosoma mansoni adult worms. Moreover, in silico studies to propose potential macromolecular targets and to predict the oral bioavailability were also performed. The analog GPQF-108 exhibited the best in vitro performance (IC50: 29.4 µM, SI:6.1) associated with promising in vivo activity, with a significant decrease in the adult life forms and oviposition. Oral bioavailability could be impaired by the predicted low water solubility of GPQF-108, although it also exhibited good membrane permeability. The water solubility, however, could be improved by decreasing the particles size. Serine/Threonine- and Tyrosine Kinases, Carbonic Anhydrase, Tyrosine Phosphatase and Arginase were predicted as potential macromolecular targets through which the GPQF-108 could be acting against the helminth. This class of compounds exhibited an interesting initial therapeutic profile with the advantage of being chemically diverse from the PZQ and be easily synthesized from commercial reagents which could lead to low-cost drugs. These aspects make this class of compounds interesting hits to be explored against schistosomiasis.

Urea’s thermal instability and burning on sensitive skin can cause problems for cosmetic formulations. To overcome these drawbacks, urea was incorporated into ordered mesoporous silica (SBA-15). SBA-15 was synthesized using tetraethyl orthosilicate and Pluronic® P123 in an acid medium. Urea (20 wt.%) was incorporated into calcined SBA-15 by the incipient wetness impregnation method. Several techniques were used to characterize the samples. Skin hydration and transepidermal water loss were measured using Corneometer® CM 825 PC and Tewameter® 300 TM. Results showed that the structural properties of SBA-15Urea were similar to pure SBA-15, indicating that SBA-15 remained structured even after urea incorporation. Nitrogen physisorption data showed the volume and surface area of the pores in SBA-15Urea were much lower than those in SBA-15, demonstrating that urea was deposited inside the mesopores. In vivo moisturization studies revealed that SBA-15Urea was not able to reduce transepidermal water loss compared to the other products and control, while forming a non-occlusive surface film on the skin. We conclude that incorporation of urea in the pores of the inorganic SBA-15 matrix is a promising approach to enhancing its stability and providing a prolonged moisturizing effect.

A biowaiver is accepted by the Brazilian Health Surveillance Agency (ANVISA) for immediate-release solid oral products containing Biopharmaceutics Classification System (BCS) class I drugs showing rapid drug dissolution. This study aimed to simulate plasma concentrations of fluconazole capsules with different dissolution profiles and run population simulation to evaluate their bioequivalence. The dissolution profiles of two batches of the reference product Zoltec® 150 mg capsules, A1 and A2, and two batches of other products (B1 and B2; C1 and C2), as well as plasma concentration–time data of the reference product from the literature, were used for the simulations. Although products C1 and C2 had drug dissolutions < 85% in 30 min at 0.1 M HCl, simulation results demonstrated that these products would show the same in vivo performance as products A1, A2, B1, and B2. Population simulation results of the ln-transformed 90% confidence interval for the ratio of Cmax and AUC0–t values for all products were within the 80–125% interval, showing to be bioequivalent. Thus, even though the in vitro dissolution behavior of products C1 and C2 was not equivalent to a rapid dissolution profile, the computer simulations proved to be an important tool to show the possibility of bioequivalence for these products.