Development Of Pharmaceutical Formulations Research Articles

Bioactive copper(II) agents and their potential involvement in the treatment of copper deficiency-related orphan diseases

The deregulation of copper homoeostasis can promote various diseases such as Menkes disease or hypertrophic cardioencephalomyopathy. We have recently synthesized solid copper(II) complexes ([Cu(His)2Cl2] and [Cu(Ser)2]), stable in physiological media and with potential as therapeutic agents. This report describes: i) the biocompatibility of these complexes at concentrations up to 100 μM using a differentiated Caco-2 cells model; ii) their transport across the intestinal epithelium using a transepithelial resistance assay and monitoring the amount of copper complexes at the apical and basolateral sides of the cells. The results suggest that the flow occurs through paracellular routes. The intracellular copper retention was <2.7% with no significant differences in intracellular copper content between 6 h and 48 h, suggesting an early copper retention process. Furthermore, this is the first evidence that demonstrates [Cu(His)2Cl2] and [Cu(Ser)2] induce transcriptional downregulation of the four major copper transporters (CTR1, DMT1, ATP7A, ATP7B), and the upregulation of the metallothionein gene expression. A remarkable finding was the increase in cytochrome c oxidase activity observed after the treatment of differentiated Caco-2 cells with copper(II) complexes at concentrations of 50–100 μM. The understanding of the transport mechanisms of these copper(II) complexes across the intestinal epithelium and of their subsequent biological activities could contribute to the development of optimal pharmaceutical formulations for the therapy of copper deficiency-related diseases.

Stability studies for the identification of critical process parameters for a pharmaceutical production of the Orf virus

A promising new vaccine platform is based on the Orf virus, a viral vector of the genus Parapoxvirus, which is currently being tested in phase I clinical trials. The application as a vaccine platform mandates a well-characterised, robust, and efficient production process. To identify critical process parameters in the production process affecting the virus’ infectivity, the Orf virus was subjected to forced degradation studies, including thermal, pH, chemical, and mechanical stress conditions. The tests indicated a robust virus infectivity within a pH range of 5–7.4 and in the presence of the tested buffering substances (TRIS, HEPES, PBS). The ionic strength up to 0.5 M had no influence on the Orf virus’ infectivity stability for NaCl and MgCl2, while NH4Cl destabilized significantly. Furthermore, short-term thermal stress of 2d up to 37 °C and repeated freeze-thaw cycles (20cycles) did not affect the virus’ infectivity. The addition of recombinant human serum albumin was found to reduce virus inactivation. Last, the Orf virus showed a low shear sensitivity induced by peristaltic pumps and mixing, but was sensitive to ultrasonication. The isoelectric point of the applied Orf virus genotype D1707-V was determined at pH3.5. The broad picture of the Orf virus’ infectivity stability against environmental parameters is an important contribution for the identification of critical process parameters for the production process, and supports the development of a stable pharmaceutical formulation. The work is specifically relevant for enveloped (large DNA) viruses, like the Orf virus and like most vectored vaccine approaches.

Advances in ATR-FTIR Spectroscopic Imaging for the Analysis of Tablet Dissolution and Drug Release.

One of the major challenges in the development of effective pharmaceutical formulations for oral administration is the poor solubility of active pharmaceutical ingredients. For this reason, the dissolution process and drug release from solid oral dosage forms, such as tablets, is usually thoroughly studied in order to understand the dissolution behaviour under various conditions and optimize the formulation accordingly. Standard dissolution tests used in the pharmaceutical industry provide information on the amount of drug released over time; however, these do not allow for a detailed analysis of the underlying chemical and physical mechanisms of tablet dissolution. FTIR spectroscopic imaging, by contrast, does offer the ability to study these processes with high spatial and chemical specificity. As such, the method allows us to see the chemical and physical processes which occur inside the tablet as it dissolves. In this review, the power of ATR-FTIR spectroscopic imaging is demonstrated by presenting a number of successful applications of this chemical imaging technique to dissolution and drug release studies for a range of different pharmaceutical formulations and study conditions. Understanding these processes is essential for the development of effective oral dosage forms and optimization of pharmaceutical formulations.

Pharmaceutical formulation design for enhancing solubility, stability, and bioavailability of poorly water-soluble drugs: An Analytical Perspective

The development of pharmaceutical formulations for drugs with poor water solubility presents significant challenges in achieving desired therapeutic outcomes. These drugs have limited solubility, which result in suboptimal bioavailability, reduced stability, and compromised efficacy. To overcome challenges, formulation strategies required to enhance drug solubility, stability, and bioavailability. Several approaches explored in pharmaceutical formulation design to address the solubility issues with poorly water-soluble drugs. These approaches particle size reduction, solid dispersion, lipid-based formulations, cyclodextrin complexation, and nanotechnology-based delivery systems. Particle size reduction techniques, such as micronization and nanonization, can significantly increase the surface area and improve dissolution rates, thereby enhancing drug solubility. Techniques such as hot melt extrusion and spray drying help disperse medications with low solubility into water-loving carriers, which boosts their solubility and ability to dissolve. Formulations that are lipid-based, like self-emulsifying drug delivery systems (SEDDS) and lipid nanoparticles, can optimize solubility and absorption of drugs by taking advantage of naturally occurring physiological pathways connected to lipid digestion and absorption. Furthermore, cyclodextrins have the capability to form complexes that aid in the increased solubility and preserving stability of medications that have little water solubility.

Oxcarbazepine: Preformulation Studies of Authoritative Part of Formulation Design

Preformulation research is a stage that is started before the novel molecule is seeded. In a larger sense, it connects with studies of a molecule's physical, chemical, analytical, and medicinal properties and offers ideas about appropriate modifications that could be made to improve performance. The analysis of preformulation characteristics can contribute to the development of pharmaceutical formulations that are efficient, safe, dependable, and stable. Oxcarbazepine is an antiepileptic drug. It controls seizures or fits by declining the abnormal and excessive activity of the nerve cells in the brain.

Development of Novel Pharmaceutical Forms of the Marine Bioactive Pigment Echinochrome A Enabling Alternative Routes of Administration.

Echinochrome A (EchA), a marine bioactive pigment isolated from various sea urchin species, is the active agent of the clinically approved drug Histochrome®. EchA is currently only available in the form of an isotonic solution of its di- and tri-sodium salts due to its poor water solubility and sensitivity to oxidation. Electrospun polymeric nanofibers have lately emerged as promising drug carriers capable of improving the dissolution and bioavailability of drugs with limited water solubility. In the current study, EchA isolated from sea urchins of the genus Diadema collected at the island of Kastellorizo was incorporated in electrospun micro-/nanofibrous matrices composed of polycaprolactone and polyvinylpyrrolidone in various combinations. The physicochemical properties of the micro-/nanofibers were characterized using SEM, FT-IR, TGA and DSC analyses. The fabricated matrices exhibited variable dissolution/release profiles of EchA, as evidenced in in vitro experiments using gastrointestinal-like fluids (pH 1.2, 4.5 and 6.8). Ex vivo permeability studies using the EchA-loaded micro-/nanofibrous matrices showed an increased permeation of EchA across the duodenum barrier. The results of our study clearly show that electrospun polymeric micro-/nanofibers represent promising carriers for the development of new pharmaceutical formulations with controlled release, as well as increased stability and solubility of EchA, suitable for oral administration, while offering the potential for targeted delivery.

Exploring in vitro solubility of lamotrigine in physiologically mimetic conditions to prospect the in vivo dissolution in pediatric population

Pediatric drugs knowledge still leaves several gaps to be filled, all the while many biopharmaceutic properties applied to adults do not work in pediatrics. The solubility in many cases is extrapolated to pediatrics; however, sometimes it may not represent the real scenario. In this context, the aim of this study was to assess the possibility of the extrapolation of the solubility data assumed for adults to children aged 2-12years using lamotrigine (LTG) as a model. LTG showed that its solubility is dependent on the pH of the medium, no precipitate formation was seen, and biomimetic media showed a greater capacity to solubilize it. Based on the dose number (D0 ) in adults, the LTG was soluble in acidic pH media and poorly soluble in neutral to basic. Similar behavior was found in conditions which mimic children aged 10-12years at a dose of 5 and 15mg/kg. The D0 for 5-year-old children at a dose of 15mg/kg showed different behaviors between biorelevant and pharmacopeial buffers media. For children aged 2-3years, LTG appeared to be poorly soluble under both gastric and intestinal conditions. Solubility was dependent on the volume of fluid calculated for each age group, and this may impact the development of better pharmaceutical formulations for this population, better pharmacokinetic predictions in tools as PBPK, and physiologically-based biopharmaceutics modeling, greater accuracy in the justifications for biowaiver, and many other possibilities.

Microparticles in the Development and Improvement of Pharmaceutical Formulations: An Analysis of In Vitro and In Vivo Studies.

Microparticulate systems such as microparticles, microspheres, microcapsules or any particle in a micrometer scale (usually of 1-1000 µm) are widely used as drug delivery systems, because they offer higher therapeutic and diagnostic performance compared to conventional drug delivery forms. These systems can be manufactured with many raw materials, especially polymers, most of which have been effective in improving the physicochemical properties and biological activities of active compounds. This review will focus on the in vivo and in vitro application in the last decade (2012 to 2022) of different active pharmaceutical ingredients microencapsulated in polymeric or lipid matrices, the main formulation factors (excipients and techniques) and mostly their biological activities, with the aim of introducing and discussing the potential applicability of microparticulate systems in the pharmaceutical field.

Physical Assessment and formulation development of Dome Matrix Controlled Release tablet of Diclofenac Sodium

In the last fifteen years where have significant acceleration in drug delivery research. In the early days of controlled release, development of pharmaceutical formulations was more idealized, with emphasis on the attainment of the illusive “zero-order release behavior”. When analyzing the swelling and release behavior of the Dome Matrix systems, it was demonstrated that the swelling behavior of the curved surfaces of the matrices strongly affected the drug release kinetics. The controlled release Dome matrix tablets containing Diclofenac sodium were prepared using a wet granulation process and polymers used xanthan gum, HPMC and Eudragit. Two different formulations were prepared; in first formulation the ratio was kept 60% active ingredient and 40% polymers subsequently in 2nd formulation 55% active ingredient 45% polymers. Precompression test angle of repose, Hausner’s ratio, Bulk Density, Tapped Density, Car’s Index were checked according to the specified standard procedure. All Physical Characteristics of dome tablets were analyzed. Drug release pattern was also analyzed by using USP apparatus II paddle method. The hardness, weight fluctuation, thickness, friability, drug content homogeneity, and in-vitro dissolution investigations of the manufactured controlled release tablets. All were examined and found within acceptable range. The F1 and F2 formulations performed well in tests and had a continuous drug release. The findings of this study showed that a polymer could be successfully used to formulate Diclofenac sodium Dome matrix tablets. The drug release was extended for 8 to 12 hours in all formulations using a drug to polymer ratio of 60:40, 55:45. The formulations are proved to have efficient drug release time. Keywords: Diclofenac sodium, Dome Matrix, Eudragit, Control release, Xanthan gum.

A material-sparing simplified buoyancy method for determining the true density of solids

True density is an important physical property of powdered materials, especially in the context of powder compaction. Currently available methods for true density determination either require a significant amount of materials or are laborious. Hence, a material-sparing and efficient method for true density determination is of value. In this work, we detail a simplified buoyancy-based method capable of fast determination of true density with accuracy comparable to helium pycnometry. This miniaturized method only uses a few milligrams of a powder with data collection process expedited by centrifugation in a laboratory centrifuge. This method can be easily adapted in a laboratory since determination of true density only requires a balance, a micropipette, a laboratory centrifuge, and standard stock liquids of low and high densities. Hence, it is a useful addition to the materials characterization tool kit critical for pharmaceutical formulation development.

Nanonized carbamazepine for nose-to-brain delivery: pharmaceutical formulation development

Epilepsy is one of the most common neurological disorders in the world. The therapeutic treatment is challenging since conventional drugs have limited efficacy and several side effects that impair patient management. Efforts are being made to find innovative strategies to control epileptic seizures. Intranasal administration provides a convenient route to deliver the drug to the brain. Carbamazepine (CBZ) is an anticonvulsant characterized by poor water solubility, nanonization can improve its bioavailability. Therefore, the design of CBZ nanocrystals (NCs) was assessed to obtain a formulation suitable for nose-to-brain delivery. CBZ NCs were prepared by sonoprecipitation following the Quality by Design approach identifying the impact of process and formulation variables on the critical quality attributes of the final product. The formulation was characterized by a technological point of view (thermotropic behavior, crystallinity, morphology, mucoadhesive strength). Response surface methodology was a reliable tool (error % 2.6) to optimize CBZ NCs with size ≤300 nm. Incubation of CBZ NCs in artificial cerebrospinal fluid at 37 °C did not promote aggregation and degradation phenomena. Preliminary biological studies revealed the biocompatibility of CBZ NCs towards Olfactory Ensheating Cells. The suspension was successfully converted into a powder. The highly concentrated formulation can be obtained, providing the possibility to administer the maximum dose of the drug in the lowest volume.

Biosynthesis of silver nanoparticles of Tribulus terrestris food supplement and evaluated antioxidant activity and collagenase, elastase and tyrosinase enzyme inhibition: In vitro and in silico approaches

Food supplements from plants are source of many antioxidant substances. Tribulus terrestris food supplement (TtFS) is highlighted as one of these supplements that has presented potential to prevent damages and diseases caused by oxidative stress. In addition, new formulations that enhance the benefits of food supplement have been developed and metallic nanoparticles, the so-called green synthesis, has been of great interest. UV–visible spectroscopy, energy dispersive spectroscopy (EDS), Zeta potential (ZP), scanning transmission electron microscopy (STEM), scanning electron microscope (SEM) were used to characterize the synthesized silver nanoparticles (AgNPs). Antioxidant activity was determined by DPPH radical scavenging, ferric ion reducing power (FRAP); ABTS radical scavenging, and oxygen radical absorption capacity (ORAC-FL). Molecular docking analyses were performed to evaluate molecular interactions. Antioxidant evaluation of TtFS demonstrated 83.6% antioxidant activity (DPPH), 714.82 μM Trolox Equivalent (TE)/g sample (FRAP), 91.24 μM TE/g sample (ABTS), and antioxidant activity in ORAC assay. It presented inhibition activity on collagenase (75.33%), elastase (54.04%) and tyrosinase (59.07%). In addition, in silico assays showed an interaction between T. terrestris steroidal saponins and DNA, and the highest DNA binding affinity was observed with protodioscin. Steroidal saponins also presented interaction with all enzymes evaluated. The characterization of AgNPs demonstrated that they were formed as a spherical shape. These AgNPs showed greater protective properties against oxidative stress (85.41% antioxidant activity (DPPH) and 957.55 μM TE/g sample (FRAP)). The results represent an important indicator for the development and discovery of new nanoestructured pharmaceutical and cosmetic formulation using plants and natural products.

Principi i primena međufazne reologije u (pre)formulacionom razvoju farmaceutskih preparata

Rheology is a science that deals with the movement and deformation of materials, while interfacial rheology is a branch of rheology that focuses on the study of the mechanical properties of fluid interfaces, such as liquid-liquid and liquid-gas interfaces. The behavior of fluid interfaces differs significantly from that of bulk materials, and standard rheological measurements of bulk materials cannot adequately describe their properties. Interfacial rheology is a specialized approach to the study of viscoelasticity and interfacial tension at liquid interfaces and provides valuable insight into intermolecular interactions and surface forces. This knowledge is particularly important for the development of advanced formulations and systems with tailored properties and functionalities. Interfacial rheology is a rapidly growing research field with diverse applications in science and industry. It plays a pivotal role in the development and optimization of pharmaceutical formulations and design of innovative drug delivery carriers with improved stability, enhanced drug release profiles, and targeted delivery capabilities. This review article aims to provide a comprehensive overview of interfacial rheology, its principles, measurement techniques, and applications in various industries. By exploring recent advancements and emerging trends in the field, we aim to highlight the significance of interfacial rheology in optimizing formulations, enhancing product performance, and driving innovations, particularly in pharmaceutical science.

Thermoanalytical and spectroscopic studies on medicated jellies with perphenazine

Medicated jellies are widely used by a large part of patients, especially by people with swallowing difficulties. Preformulation studies play an essential role in the development of new pharmaceutical formulations. The present study aimed to formulate and evaluate medicated jellies containing perphenazine, an antipsychotic drug from the group of phenothiazine compounds used to treat schizophrenia and other mental disorders. Typical gelling agents such as sodium alginate (Alg), gelatine (Gel), and pectin (Pec) were used to develop the medicated jellies. In addition to the biopolymers, components such as benzoic acid (BenzAc), citric acid (CitAc), sodium citrate (NaCit), sorbitol (Sorb), and xylitol (Xyl) were also used. Before preparing the jellies, the moist binary mixture between each component of the jelly and the active substance was analyzed to investigate the compatibility of the substances. The active substance, moist binary mixture, and medicated jellies were analyzed by FTIR_UATR spectroscopy, UV-Vis spectroscopy, and thermogravimetry (TGA).

C-glycosyl flavones in &lt;em&gt;Passiflora incarnata&lt;/em&gt;

Brazil has a diversity of plant species and many of them are medicinal plants used in the production of herbal medicines. Passiflora incarnata is popularly known as passion fruit, its fruits and aerial parts have anxiolytic and sedative properties. Recent studies have shown that P. incarnata has several bioactive compounds such as indole alkaloids, cyanogenic glycosides, maltol and flavonoids. Among the substances identified in the species, vitexin has action on the Central Nervous System, interfering with the functioning of gamma-aminobutyric acid. Thus, the objective of the study was to identify and quantify the vitexin present in the ethyl acetate fraction of the aerial parts of the passion fruit. For this, the aerial parts of P. incarnata were submitted to extraction with ethanol, followed by fractionation in organic solvents, obtaining the hexane, dichloromethane and ethyl acetate fractions. The presence of vitexin in the ethyl acetate fraction was verified by Thin Layer Chromatography (TLC) and the total flavonoid content by colorimetric reaction in a spectrophotometer. The quantification of vitexin was performed by High Performance Liquid Chromatography (HPLC). The total flavonoid content was 5.34 mg/g and 960 µg/g of vitexin in the ethyl acetate fraction. Studies like this contribute to the development of new pharmaceutical formulations from the substance vitexin.

Eugenia gracillima essential oil has pharmaceutical applications in pain and inflammation without toxic effects in mice

Ethnopharmacological relevanceEugenia gracillima is widely used by the population in the manufacture of pulps and jellies, with popular reports of its use in the treatment of infections in the urinary system, respiratory and dermatological problems. A previous study reports that EO from E. gracillima leaves proved to be a promising antioxidant agent in combating the promastigote forms of protozoa. Despite this, this species has been little studied due to its pharmacological properties. Study objectiveIn this study, an essential oil extracted (EO) from Eugenia gracillima leaves was evaluated for its acute toxicity and anti-inflammatory, antinociceptive and behavioral effects in mice. MethodsThe EO was obtained by hydrodistillation, and the composition analysis was performed by gas chromatography coupled to mass spectrometry. Acute toxicity assessment was performed with observation of hematological parameters and histopathological evaluation, as well as tests to investigate antinociceptive, anti-inflammatory activities and behavioral effects. ResultsChromatographic analysis showed D-germacrene (16.10%), γ-muurolene-g (15.60%) and bicyclogermacrene (8.53%) as the majority of compounds. In the toxicity evaluation, no death or physiological changes were observed in mice treated with a single oral dose of up to 5000 mg/kg, and it did not lyse erythrocytes in vitro. The hematological parameters evaluated were not changed after treatment; however, 5,000 mg/kg promoted an increase in transaminase levels. In the histopathological evaluation, only the animals that received the dose of 5000 mg/kg showed discrete leukocyte infiltration around the centrilobular vein in the liver. Antinociceptive activity was detected through tests of acetic acid-induced writhing, formalin, and tail flick, promoted in part by the opioid receptor pathway. In the evaluation of anti-inflammatory activity, a reduction in inflammation was observed in the paw edema test and a decrease in the migration of leukocytes and neutrophils in the peritonitis test. The open field and elevated plus maze tests showed that EO did not affect the animals' motor functions or exploratory activity. ConclusionIt was concluded that the essential oil of E. gracillima has potential for the development of pharmaceutical formulations with analgesic and anti-inflammatory actions in non-toxic concentrations.

Neither too little nor too much: Finding the ideal proportion of excipients using confocal Raman and chemometrics

The applications of Raman imaging in pharmaceutical field are ever-increasing due its ability to obtain spatial and spectral information simultaneously, once it allows determine the chemical distribution of compounds. In this sense, it is used to study homogeneity, of paramount importance during the development of pharmaceutical formulations due to its relation to stability, safety and efficacy. Commonly, just surface is analyzed, but confocal Raman spectroscopy can also characterize the inner part of samples, allowing to determine phase separation in the early stages. In this sense, confocal 3D Raman microscopy was crucial to obtain the optimal proportion of Apifil®, Capryol® 90 and Transcutol® to promote controlled release of the local anesthetic butamben (BTB). 3D chemical maps were obtained by classical least squares (CLS) using pure compound spectra as S matrix, showing that chemical distribution throughout the material was different. Knowing that the composition of samples affects the homogeneity parameter, standard deviation and distributional homogeneity index (DHI) were used in mixture experimental design (DoE). From this analysis, it was revealed that a correct amount of Capryol® 90 enhances both miscibility and solubility. Furthermore, suitable miscibility was observed in two ratio proportions of excipients with a desirability of 0.783 and 0.742. These results unequivocally demonstrated that confocal Raman microscopy combined to DoE can bring pharmaceutical development to a higher level.

Emerging Artificial Intelligence (AI) Technologies Used in the Development of Solid Dosage Forms.

Artificial Intelligence (AI)-based formulation development is a promising approach for facilitating the drug product development process. AI is a versatile tool that contains multiple algorithms that can be applied in various circumstances. Solid dosage forms, represented by tablets, capsules, powder, granules, etc., are among the most widely used administration methods. During the product development process, multiple factors including critical material attributes (CMAs) and processing parameters can affect product properties, such as dissolution rates, physical and chemical stabilities, particle size distribution, and the aerosol performance of the dry powder. However, the conventional trial-and-error approach for product development is inefficient, laborious, and time-consuming. AI has been recently recognized as an emerging and cutting-edge tool for pharmaceutical formulation development which has gained much attention. This review provides the following insights: (1) a general introduction of AI in the pharmaceutical sciences and principal guidance from the regulatory agencies, (2) approaches to generating a database for solid dosage formulations, (3) insight on data preparation and processing, (4) a brief introduction to and comparisons of AI algorithms, and (5) information on applications and case studies of AI as applied to solid dosage forms. In addition, the powerful technique known as deep learning-based image analytics will be discussed along with its pharmaceutical applications. By applying emerging AI technology, scientists and researchers can better understand and predict the properties of drug formulations to facilitate more efficient drug product development processes.

Tertiary Nanosystem Composed of Graphene Quantum Dots, Levofloxacin and Silver Nitrate for Microbiological Control.

Infectious diseases have the highest mortality rate in the world and these numbers are associated with scarce and/or ineffective diagnosis and bacterial resistance. Currently, with the development of new pharmaceutical formulations, nanotechnology is gaining prominence. Nanomicelles were produced by ultrasonication. The particle size and shape were evaluated by scanning electron microscopy and confirmed by dynamic light scattering, also thermogravimetric analysis was performed to evaluate the thermal stability. Finally, antibacterial activity has been performed. The results showed that a rod-shaped nanosystem, with 316.1 nm and PDI of 0.243 was formed. The nanosystem was efficient against Staphylococcus aureus, Pseudomonas aeruginosa, and Bacillus subtilis subsp. spizizenii with MIC inferior to 0.98 and a synergistic effect between silver graphene quantum dots and levofloxacin was observed. The nanosystem produced may rise as a promising agent against the bacterial threat, especially regarding bacterial resistance.

In vitro effect on the proteolytic activity of papain with proteins of the skin as substrate.

This work aims to evaluate the effects of enzyme concentration, pH, temperature and time course degree of hydrolysis (DH) of papain regarding further development of pharmaceutical and cosmetic formulations. The hydrolysis of casein, collagen, keratin and porcine skin at pH and temperature ranges of the human skin was evaluated. Also, low contact times of enzyme-substrate were studied. The incorporation of 3mM of cysteine improved the caseinolytic (PU), collagenolytic (CU) and keratinolytic (KU) activities of papain. In general, the increase from 0.1 to 1.0 or 2.0mgmL-1 of papain improved PU, CU and KU. When 2.0mgmL-1 of papain was used, the highest DH of casein, collagen and keratin was obtained at 240min (14%, 35% and 6% respectively). The decrease in pH and temperature reduced all proteolytic activities, but papain maintained at least 50% and 40% of its activity at 26°C and pH 4.5 respectively. Scanning electron micrographs of the surface of the skin showed that papain application had exfoliating activity. This preformulation study demonstrated that papain concentration, time of application and pH of the product should be evaluated when developing a product to promote the hydrolysis of the proteins of the skin.