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Forarticles published under an open access Creative Common CC BY license, any part of the article may be reused withoutpermission provided that the original article is clearly cited.Feature Papers represent the most advanced research with significant potential for high impact in the field. FeaturePapers are submitted upon individual invitation or recommendation by the scientific editors and undergo peer reviewprior to publication.The Feature Paper can be either an original research article, a substantial novel research study that often involvesseveral techniques or approaches, or a comprehensive review paper with concise and precise updates on the latestprogress in the field that systematically reviews the most exciting advances in scientific literature. This type ofpaper provides an outlook on future directions of research or possible applications.Editor’s Choice articles are based on recommendations by the scientific editors of MDPI journals from around the world.Editors select a small number of articles recently published in the journal that they believe will be particularlyinteresting to authors, or important in this field. The aim is to provide a snapshot of some of the most exciting workpublished in the various research areas of the journal. You seem to have javascript disabled. Please note that many of the page functionalities won't work as expected without javascript enabled. clearsearchmenuJournalsTopicsInformationFor AuthorsFor ReviewersFor EditorsFor LibrariansFor Publishers For SocietiesFor Conference OrganizersOpen Access PolicyInstitutional Open Access Program Special Issues GuidelinesEditorial ProcessResearch and Publication EthicsArticle Processing Charges AwardsAuthor ServicesInitiatives SciforumMDPI BooksPreprintsScilitSciProfiles EncyclopediaJAMSProceedings Series AboutOverviewContactCareers Sign In / Sign UpSubmit5.3JournalsSci. Pharm.Journal DescriptionScientia PharmaceuticaScientia Pharmaceuticais an international,peer-reviewed, open access journal related to the pharmaceutical sciences. It is the journal of the Austrian Pharmaceutical Society (Österreichische Pharmazeutische Gesellschaft, ÖPhG) and it is published quarterly online by MDPI, and in print by the Austrian Pharmacists' Publishing House (Österreichischer Apothekerverlag).Open Access— free for readers, witharticle processing charges (APC) paid by authors or their institutions.High Visibility: indexedwithin Scopus, ESCI (Web of Science), Embase, CAPlus / SciFinder,and manyother databases.Journal Rank:CiteScore- Q2 (Pharmaceutical Science)Rapid Publication: manuscripts are peer-reviewed and a firstdecision provided to authors approximately 18.8 days after submission; acceptanceto publication is undertaken in 4.9 days (median values for papers published inthis journal in the second half of 2021).Recognition of Reviewers: reviewers who provide timely, thorough peer-review reports receive vouchers entitling them to a discount on the APC of their next publication in any MDPI journal, in appreciation of the work done.subjectImprint Informationget_appJournal Flyer Open Access ISSN: 2218-0532Latest ArticlesattachmentSupplementary material:Supplementary File 1 (ZIP, 291 KiB)get_appsubjectView online as:Abstract Page Full-Text HTMLOpen AccessArticleIn Vitro and In Silico Antistaphylococcal Activity of Indole Alkaloids Isolated from Tabernaemontana cymosa Jacq (Apocynaceae)byYina Pájaro-González, Julián Cabrera-Barraza, Geraldine Martelo-Ramírez, Andrés F. Oliveros-Díaz, Juan Urrego-Álvarez, Wiston Quiñones-Fletcher and Fredyc Díaz-Castillo Sci. Pharm. 2022, 90(2), 38; https://doi.org/10.3390/scipharm90020038 - 14 Jun 2022Abstract The species of the genus Tabernaemontana have a long tradition of use in different pathologies of infectious origins; the antibacterial, antifungal, and antiviral effects related to the control of the pathologies where the species of this genus are used, have been attributed to [...] Read more.The species of the genus Tabernaemontana have a long tradition of use in different pathologies of infectious origins; the antibacterial, antifungal, and antiviral effects related to the control of the pathologies where the species of this genus are used, have been attributed to the indole monoterpene alkaloids, mainly those of the iboga type. There are more than 1000 alkaloids isolated from different species of Tabernaemontana and other genera of the Apocynaceae family, several of which lack studies related to antibacterial activity. In the present study, four monoterpene indole alkaloids were isolated from the seeds of the species Tabernaemontana cymosa Jacq, namely voacangine (1), voacangine-7-hydroxyindolenine (2), 3-oxovoacangine (3), and rupicoline (4), which were tested in an in vitro antibacterial activity study against the bacteria S. aureus, sensitive and resistant to methicillin, and classified by the World Health Organization as critical for the investigation of new antibiotics. Of the four alkaloids tested, only voacangine was active against S. aureus, with an MIC of 50 µg/mL. In addition, an in silico study was carried out between the four isolated alkaloids and some proteins of this bacterium, finding that voacangine also showed binding to proteins involved in cell wall synthesis, mainly PBP2 and PBP2a.Full article(This article belongs to the Special Issue Feature Papers in Scientia Pharmaceutica)►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 Chemical structures of indole alkaloids from Tabernaemontana cymosa. Full article ">Figure 2 Heat-map of the molecular docking affinities value (kcal/mol) for T. cymosa alkaloids with S. aureus proteins. Full article ">Figure 3 Molecular interactions between voacangine and PBP2 from S. aureus. (A) Characterization of the binding site according to the ability to form hydrogen bridges and geometry of the compound at the docking site, (B) 2D plot of the interactions specifying the type of interaction, (C) conformation of the compound during docking and interactions with PBP2 residues in 3D. Full article ">Figure 4 Molecular interactions between voacangine PBP2a and S. aureus. (A) Characterization of the binding site according to the ability to form hydrogen bridges and geometry of the compound at the docking site, (B) 2D plot of the interactions specifying the type of interaction, (C) conformation of the compound during docking and interactions with PBP2a residues in 3D. Full article "> attachmentSupplementary material:Supplementary File 1 (ZIP, 638 KiB)get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleEx Vivo and In Vivo Study of Some Isoquinoline PrecursorsbyMiglena Milusheva, Vera Gledacheva, Margarita Batmazyan, Stoyanka Nikolova, Iliyana Stefanova, Darinka Dimitrova, Kremena Saracheva, Desislav Tomov and Veneta Chaova-Gizdakova Sci. Pharm. 2022, 90(2), 37; https://doi.org/10.3390/scipharm90020037 - 13 Jun 2022Abstract This article concerns the synthesis and biological activities of some N-(1-(3,4-dimethoxyphenyl)propan-2-yl) amides as isoquinoline precursors and compounds with smooth muscle (SM) relaxant activity. Aim: find the biological activity of N-(1-(3,4-dimethoxyphenyl)propan-2-yl) amides and compare it with papaverine, an isoquinoline alkaloid that has been known [...] Read more.This article concerns the synthesis and biological activities of some N-(1-(3,4-dimethoxyphenyl)propan-2-yl) amides as isoquinoline precursors and compounds with smooth muscle (SM) relaxant activity. Aim: find the biological activity of N-(1-(3,4-dimethoxyphenyl)propan-2-yl) amides and compare it with papaverine, an isoquinoline alkaloid that has been known as a brain and coronary vasodilator and SM relaxant. Materials and methods: In silico simulation with the PASS online program predicts SM relaxant activity for the compounds. The amides were tested on the isolated gastric SM preparations (SMPs) from rats to determine their effects on spontaneous contractile activity (CA) compared with papaverine. The in vivo effect on the learning and memory processes of rats was also assessed. Results: the data from the isometric measurements showed that one of the compounds caused ex vivo relaxation in circular SM tissues isolated from the stomach (corpus) of male Wistar rats. Conclusion: We found that the compound’s SM relaxation uses the papaverine pathway. It also has an improving effect on the cognitive functions of learning and memory processes in rats.Full article(This article belongs to the Special Issue Feature Papers in Scientia Pharmaceutica)►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 Chemical structure of papaverine. Full article ">Figure 2 Relaxation effects caused by 1 × 10−6 mol/L papaverine and 5 × 10−5 mol/L 4d on SMPs at single or combined administration (n = 12); * p < 0.05. Full article ">Figure 3 Representative tracings of: (a) corpus SMP from rat elicited by 1 × 10−6 mol/L papaverine and 5 × 10−5 mol/L 4d at single or combined administration; (b) SM activity parameters before and after KCl depolarization. Full article ">Figure 4 Effects of 4d on learning and memory processes in rats (shuttle-box active avoidance test, number of avoidances); 0 p < 0.05 compared with day 1 control group; * p < 0.05 compared with the same-day control group. Full article ">Figure 5 Effects of 4d on learning and memory processes in rats (shuttle-box active avoidance test, number of escapes) * p < 0.05 compared with the same-day control group. Full article ">Figure 6 Effects of 4d on learning and memory processes in rats (shuttle-box active avoidance test, number of intertrial crossings) * p < 0.05 compared with the same-day control group; ** p < 0.01 compared with the same-day control group. Full article ">Figure 7 Effects of 4d on learning and memory processes in rats (step-down passive avoidance test); 0 p < 0.05 compared with day 1 control group; * p < 0.05 compared with the same-day control group. Full article ">Scheme 1 Synthesis of 1-(3,4-dimethoxyphenyl)propan-2-amine 3. Full article ">Scheme 2 Synthesis of N-(1-(3,4-dimethoxyphenyl)propan-2-yl) amides 4. Full article "> get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleOptimized Methods for Analytical and Functional Comparison of Biosimilar mAb Drugs: A Case Study for Avastin, Mvasi, and ZirabevbyBüşra Gürel, Eda Çapkın, Ayhan Parlar, Aylin Özkan, Meltem Çorbacıoğlu, Duygu Emine Dağlikoca and Meral Yüce Sci. Pharm. 2022, 90(2), 36; https://doi.org/10.3390/scipharm90020036 - 31 May 2022Abstract Bevacizumab is a humanized therapeutic monoclonal antibody used to reduce angiogenesis, a hallmark of cancer, by binding to VEGF-A. Many pharmaceutical companies have developed biosimilars of Bevacizumab in the last decade. The official reports provided by the FDA and EMA summarize the analytical [...] Read more.Bevacizumab is a humanized therapeutic monoclonal antibody used to reduce angiogenesis, a hallmark of cancer, by binding to VEGF-A. Many pharmaceutical companies have developed biosimilars of Bevacizumab in the last decade. The official reports provided by the FDA and EMA summarize the analytical performance of biosimilars as compared to the originators without giving detailed analytical procedures. In the current study, several key methods were optimized and reported for analytical and functional comparison of bevacizumab originators (Avastin, Altuzan) and approved commercial biosimilars (Zirabev and Mvasi). This case study presents a comparative analysis of a set of biosimilars under optimized analytical conditions for the first time in the literature. The chemical structure of all products was analyzed at intact protein and peptide levels by high-resolution mass spectrometry; the major glycoforms and posttranslational modifications, including oxidation, deamidation, N-terminal PyroGlu addition, and C-terminal Lys clipping, were compared. The SPR technique was used to reveal antigen and some receptor binding kinetics of all products, and the ELISA technique was used for C1q binding affinity analysis. Finally, the inhibition performance of the samples was evaluated by an MTS-based proliferation assay in vitro. Major glycoforms were similar, with minor differences among the samples. Posttranslational modifications, except C-terminal Lys, were determined similarly, while unclipped Lys percentage was higher in Zirabev. The binding kinetics for VEGF, FcRn, FcγRIa, and C1q were similar or in the value range of originators. The anti-proliferative effect of Zirabev was slightly higher than the originators and Mvasi. The analysis of biosimilars under the same conditions could provide a new aspect to the literature in terms of the applied analytical techniques. Further studies in this field would be helpful to better understand the inter-comparability of the biosimilars.Full article►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 Intact Protein Analysis of AVT, ALT, ZIR, and MVA. (A) Schematic illustration of bevacizumab glycosylation site and main glycans. (B) Overlay representation of UV chromatograms of samples. Full article ">Figure 2 Comparative raw MS spectrums of Zirabev (A) and Mvasi (B) in zoom-out(up) and zoom-in(down) form. (C) Overlay representation of deconvoluted MS spectrums of samples and the list of observed molecular mass of the main peaks. (D) Graph of percentages of major glycoforms identified in the samples. Each sample was injected three times. Full article ">Figure 3 Peptide mapping analysis of the samples. (A) Comparison of peptide profiles of samples by overlaying TIC chromatograms of tryptic peptides. (B) A representative coverage map of ALT indicating 99% coverage. The blue highlighting indicates the identified sequences. Full article ">Figure 4 Peptide mapping analysis of the samples. (A) The schematic representations of Bevacizumab and approximate locations of modified sequences. (B) The table of complementarity-determining regions (CDRs) in heavy and light chains, their sequence and positions, and retention time observed. (C) The table of modified peptides, modifications, and relative content. All samples were injected three times, and the values in the table are the average values. ND not detected. (*) represents carbamidomethyl-C modification. Bold and underlined letters indicate the CDR sequences, while red letters indicate modified amino acid residues. Full article ">Figure 5 VEGF binding analysis of the samples (A) Specific interaction of bevacizumab-based monoclonal antibody drugs with free VEGF molecules in the extracellular region inhibits angiogenesis. (B) SPR results for VEGF binding. The data represented the mean of KD values obtained from at least 3 independent measurements. (C) Results of equivalency test. The Upper and lower limit was determined as mean ± 1.5 × SD. Full article ">Figure 6 FcRn binding analysis of the samples (A) Typical sensograms for an injected mAb sample and immobilized FcRn were shown. FcRn captures mAb molecules at acidic pH and releases them when the pH becomes neutral. (B) Steady-state interaction of bevacizumab-based mAb-based samples with immobilized FcRn molecules was represented as the mean of at least three measurements. (B) Langmuir 1:1 binding model based SPR results for VEGF binding. The data represented the mean of at least three independent measurements. (C) Two-state binding interaction of bevacizumab-based mAb-based samples with immobilized FcRn molecules was represented as the mean of at least three measurements. There was no significant difference between Avastin, and other samples based on the Single-way ANOVA analysis (p < 0.05). Full article ">Figure 7 FcγRIa binding analysis of Bevacizumab and biosimilars. (A) Steady-state interaction of the samples with the Anti-His capture method was represented as the mean of at least three measurements. (B) Langmuir 1:1 binding model based SPR results for FcγRIa binding. The data represented the mean of at least three independent measurements. There was no significant difference between Avastin, and other samples based on the Single-way ANOVA analysis (p < 0.05). Full article ">Figure 8 C1q binding analysis of the samples. (A) Schematic illustration of C1q complex (left) and basic C1q and IgG interaction (right). (B) The results of C1q binding of AVT, ALT, ZIR, and MVA by ELISA at different concentrations. The data represented the mean of at least three independent measurements. There was no significant difference between Avastin, and other samples based on the statistical analysis (p < 0.05). Full article ">Figure 9 Inhibitory effect of samples on VEGF-induced HUVEC proliferation. (A) Schematic illustration of the action mechanism of anti-VEGF antibody. (B) Comparison of the potency of originators and biosimilars was represented as relative proliferation (%). Each sample was analyzed in triplicate. (C) Equivalence test results represent the differences in means of biosimilars comparing the originators. Full article "> attachmentSupplementary material:Supplementary File 1 (ZIP, 392 KiB)get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleConcepts for New Rapid Simple HPLC Method for Quantification of Fosfomycin Trometamol in Pharmaceutical Dosage Forms with Direct UV DetectionbyMarjan Piponski, Tanja Bakovska Stoimenova, Tetiana Melnyk, Sergiy Kovalenko, Elena Lazarevska Todevska, Marjan Velkovski, Sami El Deeb, Yuriy Mysula and Liliya Logoyda Sci. Pharm. 2022, 90(2), 35; https://doi.org/10.3390/scipharm90020035 - 24 May 2022Abstract Two different concepts for developing direct HPLC-UV methods for quantifying fosfomycin trometamol were developed without any derivatization and modification of the analyte. In the first concept, without the use of alkylamines as ion-pairs in the mobile phase, by using cyanopropyl CN and a [...] Read more.Two different concepts for developing direct HPLC-UV methods for quantifying fosfomycin trometamol were developed without any derivatization and modification of the analyte. In the first concept, without the use of alkylamines as ion-pairs in the mobile phase, by using cyanopropyl CN and a strong anion-exchanger column, we investigated the possibility of their highly polar and anion-exchanging forces and mechanisms to retain, separate and detect trometamol without the help of additional agents or modifiers. In the second concept, the most frequent reversed-phase C18 columns with different characteristics and vendors were tested in combination with different length-based alkylamines with 3–10 C atoms in their chains. In our research, we found that the ion-pairing of fosfomycin with 6–10 C-atom-based alkyl-length of aliphatic chains manifested the most appropriate strength of interactions between alkyl-paired trometamol molecules and octadecylsilane or C18 bonded RP column to achieve optimal retention, selectivity and peak shape on chromatograms, with the possibility for the fine-tuning of elution time. The simplicity of our method concept omits the need for expensive and sophisticated columns like HILIC, C30 graphite carbon, and mixed-mode-based columns for easier retaining, separation, and determination of fosfomycin, and for its quantification purposes, especially in high-throughput analyses in regular quality-control laboratories.Full article►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 (A) Illustration of one of the drawbacks of a NH2- column, including a long equilibration resulting in long-lasting baseline shifts, easily induced with the smallest change of T in less than 0.2 °C change, smallest variations of flow rate, mobile phase refill, and injection amount of analyte with calculated peak symmetry of fosfomycin of 3.722. (B) Chromatogram of obtained HPLC-NH2-RID analyses according to pharmacopoeia with an injected 10 μL 120 mg/mL, or 1.2 mg on column with a standard solution of fosfomycin trometamol (0.463 mol/L). Full article ">Figure 2 UV-Vis absorption spectrum of 0.1 mg/mL pure standard of fosfomycin trometamol in water; the blue-lined spectrum was overlapped with and masked by trometamine residue. Full article ">Figure 3 Influence of pH of the mobile phase buffer on the retention and selectivity of fosfomycin trometamol with an Agilent Zorbax SB-CN column with potassium phosphate buffers with different levels of acidity. Lowering the pH resulted in inducing a separation of the analyte component mixture of 24 mg/mL standard of fosfomycin trometamol in the mobile phase, eluting at 2.77 min. Full article ">Figure 4 Chromatogram obtained by injection of 24 mg/mL standard on Zorbax SB-CN 250 mm × 4.6 mm column at 192 nm and 24 and 48 mg/mL standard of fosfomycin eluting at 2.781 min. Mobile phase composed of 0.55% w/v KH2PO4 and 0.5% acetinitrile pH = 2.45, 1 mL/min flow rate, injection volume—1 and 2 μL of 24 mg/mL standard fosfomycin trometamol dissolved in the mobile phase at UV = 195 nm and 35 °C. Full article ">Figure 5 Chromatogram gained with 0.5% w/v NaH2PO4 pH = 2.5 + 0.5% acetonitrile with 1 mL/min (left part) and chromatogram with a 3× diluted mobile phase with water and 0.55 mL/min (right part). Full article ">Figure 6 Chromatogram with LiChrospher-CN column with fosfomycin peak eluting at about 2.75 min; overlayed signals from 190 nm + 195 nm for direct comparison with injected 5 μL standard with 24 mg/mL fosfomycin trometamol in the mobile phase with 0.05% w/v KH2PO4 + 0.2% acetonitrile. The pH was adjusted to 2.4 with the use of a Shimadzu Nexera LC-2040c 3-D. Full article ">Figure 7 Chromatograms at two wavelengths for a comparison of the selectivity of the cyanopropyl column, LiChrospher-CN 250 mm × 4 mm, in the quantification of old granulate with a standard of fosfomycin. The degradation product increment is easily visible—26 mg/mL standard with 26.4 mg/mL granulate. A mobile phase of 0.65% KH2PO4 pH = 2.36 and 0.15% acetonitrile, 5 μL injection volume, UV − 190 nm + 195 nm, 1 mL/min, with the use of a Shimadzu Nexera LC-2040c 3-D2. Full article ">Figure 8 Chromatogram of third cyanopropyl type, Waters Spherisorb SCN column, 2 and 3 μL injection volume at UV-192 nm, resulting in a retention of fosfomycin at tR = 2.86 min with t5% = 1.59. Full article ">Figure 9 Anion-exchange column with a Partisil SAX column 250 mm × 4.6 mm, 10 μm particles at UV = 192 nm with an older type of HPLC Varian ProStar PDA 330, with 50 mM NH4H2PO4 and 5% acetonitrile at 35 °C and flow rate 1 mL/min. Fosfomycin with 50 mg/mL and 5 μL injection volume at 195 nm, chromatograph and controlled with Varian Star software version 6.30. Full article ">Figure 10 Chromatogram of fosfomycin trometamol on Whatman Partisil SAX 250 mm × 4.6 mm, 10 μm column, on a Shimadzu Nexera LC-40 3D Plus. The chromatogram, contour map, diagram and 3-D UV spectrum, UV absorption spectra of the main two peaks with system suitability parameters are noted below in the table. injection volume of 1 μL with standard 32 mg/mL of fosfomycin in the mobile phase. Full article ">Figure 11 Chromatogram of fosfomycin trometamol on a Whatman Partisil SAX 250 mm × 4.6 mm, 10 μm column, on Shimadzu Nexera LC-40 3D Plus illustrating the chromatogram, contour map, diagram and 3-D UV spectrum and peaks of the UV absorption spectra of the main two peaks. The system suitability parameters are below in the table. An injection volume of 1 μL with a standard 32 mg/mL of fosfomycin in the mobile phase (top) and chromatogram of fosfomycin on a Whatman Partisil SAX column 250 mm × 4.6 mm, 10 μm, with a mobile phase composed of 5% acetonitrile + 95% of 0.57% NH4H2PO4 and injection of 1 μL standard solution of 32 mg/mL fosfomycin trometamol, at 1 mL/min and compared peak parameters values of two wavelengths, 190 nm + 195 nm of UV-absorption (bottom), with the illustrated chromatogram at 190 nm. Full article ">Figure 12 Chromatograms illustrating linearity of the method using column 150 mm × 4.6 mm Discovery C18 and diisopropyl amine for increase or retention of fosfomycin, monitored at UV 192 nm signal, 1 mL/min 30 °C, with 5% acetonitrile and 95% of 0.2% v/v diisopropylamine at pH = 6, without phosphate salts as buffers using a Varian ProSar330 HPLC system. Full article ">Figure 13 Chromatograms of a standard solution of fosfomycin trometamol 10.3 mg/mL on a Discovery C18 150 mm × 4.6 mm and 5 μm column with a mobile phase with 7% acetonitrile, 93% of 10 mM NH4H2PO4 and 0.21% hexylamine, pH = 6.1, flow rate 1 mL/min, two wavelengths at 192 nm and 195 nm for comparison of sensitivity with the use of Varian ProSar330 HPLC system. Full article ">Figure 14 Chromatograms of the standard solution of fosfomycin trometamol 10.3 mg/mL, on the Discovery C18 150 mm × 4.6 mm, 5 μm column with mobile phase with 6% acetonitrile, 94% of 10 mM NH4H2PO4 and 0.21% hexylamine, pH = 6.1, flow rate 1 mL/min, two wavelengths 190 nm and 195 nm, for comparison of sensitivity, worked on Shimadzu Nexera LC-40. Full article ">Figure 15 Comparison of elution profiles of two different amines, dibutyalamine DBA and hexylamine, used in slightly different organic acetonitriles with described mobile phase composition on the chromatogram apex of fosfomycin, with system suitability parameters described in the table. Full article ">Figure 16 Chromatograms of a standard solution of fosfomycin trometamol 30 mg/mL, on the Discovery C18 150 mm × 4.6 mm, 5 μm column, with the mobile phase with mixed 6% acetonitrile, 94% of 10 mM KH2PO4 and 0.2% hexylamine, pH = 6.1, at 195 nm, for comparison of sensitivity, worked on Dionex Ultimate 3000. Full article ">Figure 17 Chromatogram of the fosfomycin trometamol standard 20 mg/mL injection volume 2 μL, separated on Discovery C18 150 mm × 4.6 mm, 5 μm column with the use of 2-aminoheptan as the ion-pairing reagent. Full article ">Figure 18 Chromatograms of selectivity of method confirmation with the usage of Zorbax C18 extend column and mobile phase composed of mixed 91% of 25 mM 2-aminoheptane pH 5.8, 9% acetonitrile, sample standard concentration of 11 mg/mL, injection volume 2 uL, measured at 190 nm and 195 nm at 35 °C, with UV-Spectrum of fosfomycin. Full article ">Figure 19 Chromatograms PDA extractions overlaid at 11 wavelengths with an increment of 1 nm for comparison of change of peaks size behavior, and especially of fosfomycin. Full article ">Figure 20 Chromatograms presenting the linearity of the method with 25 mM 2-aminoheptane and Zorbax C18 extent column in the range of 0.11–55 mg/mL of fosfomycin trometamol and monitored at a 190 nm UV-absorbing signal. Mobile phase composed of mixed 91% of 25 mM 2-aminoheptane pH 5.8, 9% acetonitrile. Full article ">Figure 21 (1) Sample of forced degradation of standard fosfomycin; (2) standard of fosfomycin trometamol; (3) API standard trometamol; (4) comparative limiting 0.3% standard of fosfomycin trometamol with 2 μL injections at 190 nm UV monitoring. Full article ">Figure 22 Chromatograms of a sample of 21 mg/mL of fosfomycin trometamol with 150 mm × 4.6 mm, 3 μm YMC C18 Triart, worked with the mobile phase with 13% acetonitrile and 0.25% 2-aminoheptane, pH = 6. Full article ">Figure 23 Chromatograms with YMC C18- Triart 150 mm × 4.6 mm, 3 μm with injected 1 μL sample substance FSF with 21 mg/mL and mobile phase with 0.28% v/v—Heptylamine pH adjusted to 6 and +10%Acn. Full article ">Figure 24 Fosfomycin peak tailing and height/area ratio versus the quantity of injected analyte on the column at a UV 190 nm signal monitoring of the Waters Xterra C18 MS 250 mm × 4.6 mm, 5 μm column. Full article ">Figure 25 Chromatograms of 14 mg/mL solution of fosfomycin trometamol on a Waters Xterra C18 MS 250 mm × 4.6 mm, 5 μm column with the usage of 0.25% v/v Hexylamine as the ion-pairing amine adjusted to pH 6 with 85% o-H3PO4 and added 7% acetonitrile at 40 °C, 1 mL/min flow rate. Full article "> get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessReviewQuality by Design: A Suitable Methodology in Industrial Pharmacy for Costa Rican UniversitiesbyLuis Castillo-Henríquez, Brayan Murillo-Castillo, Lexi Chaves-Siles, Juan José Mora-Román, Nils Ramírez-Arguedas, Édgar Hernández-Mora and José Vega-Baudrit Sci. Pharm. 2022, 90(2), 34; https://doi.org/10.3390/scipharm90020034 - 19 May 2022Abstract This review aims to present the Quality by Design (QbD) model as a suitable methodology to perform research in the academic Costa Rican institutions that teach Pharmacy. Pubmed, Science Direct, and Google Scholar databases were screened for original research papers and review papers [...] Read more.This review aims to present the Quality by Design (QbD) model as a suitable methodology to perform research in the academic Costa Rican institutions that teach Pharmacy. Pubmed, Science Direct, and Google Scholar databases were screened for original research papers and review papers published not more than ten years ago. Institutional repositories from the different universities were reviewed as well. The QbD model stands out as a great methodology for carrying out research projects regarding Pharmaceutical Sciences, but especially for Industrial Pharmacy, where it has contributed in terms of formulation development, manufacturing, and quality control. Academic research based on this model enables the training and development of practical, scientific, and leadership skills in Industrial Pharmacy students. The generated knowledge can be shared in classrooms, which represents an ideal environment to communicate research results and to foster collaborative work between researchers, professors, and students. Moreover, research performed through a QbD approach increases the confidence shown by the industrial sector and health regulatory authorities in the quality of the research, products, and knowledge that are developed and created in an Academy. As a result, the implementation of the model has allowed the creation, transfer, and materialization of knowledge from the Costa Rican Academy to different local pharmaceutical industries.Full article►▼Show Figuresdiv" data-cycle-log="false">Graphical abstractGraphical abstractFull article ">Figure 1 QbD lifecycle. Reprinted with permission from Fornaguera. et al. Journal of Personalized Medicine, 7(4). Copyright (2017) MDPI [40]. Full article ">Figure 2 Example of an Ishikawa diagram for risk management in Formulation Development carried out in the Academy. Reprinted with permission from Castillo, L. et al. Drug Development and Industrial Pharmacy, 45(10). Copyright (2019) Taylor & Francis [44]. Full article ">Figure 3 Example of a Design of Experiments carried out in the Academy to assess the performance of solid formulations in the dissolution test. Adapted with permission from Castillo, L. et al. Journal of Drug Delivery and Therapeutics, 9(1-s). Copyright (2019) JDDT [54]. Full article ">Figure 4 Potential applications of the QbD approach in diverse stages of the Drug development lifecycle. Reprinted with permission from Rahman, M. et al. European Pharmaceutical Review, 22(1). Copyright (2017) Rusell Publishing Limited [39]. Full article ">Figure 5 Traditional and AQbD approaches for Analytical Method Development. Reprinted with permission from Raman, N. et al. Journal of Chemistry, 2015. Copyright (2015) Hindawi [100]. Full article ">Figure 6 Example of a 32 full factorial design carried out in the Academy to evaluate the moisture content (%) in natural raw materials as a function of the balance’s temperature (°C) and sample’s mass (g): (a) Camelia sinensis, (b) Cassia fistula, (c) Chamaemelum nobile, (d) Lippia alba, and (e) Tilia platyphyllos. Reprinted with permission from Castillo, L. et al. Borneo Journal of Pharmacy, 3(1). Copyright (2020) Institute for Research and Community Services Universitas Muhammadiyah Palangkaraya [105]. Full article "> get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleQuality and In Vivo Assessment of a Fulvic Acid Complex: A Validation StudybyRahmuddin Khan, Pooja Jain, Foziyah Zakir, Mohd Aqil, Sameer Alshehri, Mohd Aamir Mirza and Zeenat Iqbal Sci. Pharm. 2022, 90(2), 33; https://doi.org/10.3390/scipharm90020033 - 19 May 2022Abstract The present work aimed to re-assess the bioavailability enhancement potential of fulvic acid (FA). Carbamazepine (CBZ) and peat were used as a model drug and FA source, respectively. Our group has already evaluated the bioavailability enhancement potential of a less commercially viable source [...] Read more.The present work aimed to re-assess the bioavailability enhancement potential of fulvic acid (FA). Carbamazepine (CBZ) and peat were used as a model drug and FA source, respectively. Our group has already evaluated the bioavailability enhancement potential of a less commercially viable source of FA, i.e., shilajit. In the present work, the phase solubility of CBZ was analyzed with varying concentrations of peat-sourced FA (2–12% w/v). The prepared complex (CBZ-FA) was characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and differential scanning calorimetry (DSC). Dissolution, pharmacokinetic, and pharmacodynamic studies were also carried out. The results showed the presence of an interaction between the drug and FA within the complex, which led to 98.99 ± 2.0% enhancement in drug solubility. The results also showed 79.23 ± 2.1% dissolution of the complexed drug over 60 min and 69.32 ± 2.2% permeation from the intestinal gut sac over 90 min, which led to a significant enhancement of bioavailability and a reduction in the duration of epileptic seizures. Thus, this study re-authenticates our earlier results and suggests switching the FA source (shilajit to peat) for commercial product development.Full article►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 Phase solubility diagram of FA and CBZ at room temperature (25 ± 2 °C). Full article ">Figure 2 FTIR overlay spectra of FA (A), CBZ (B), and CBZ-FA complex (C). Full article ">Figure 3 Overlay of DSC thermograms of (A) CBZ, (B) FA, and (C) CBZ-FA complex. Full article ">Figure 4 Overlay of X-ray diffractograms of A (FA), B (CBZ), and C (CBZ-FA complex). Full article ">Figure 5 Comparative release profile of the pure drug (FA) and the complexed drug (CBZ-FA) in simulated gastric fluid, pH 1.2 at 37 ± 2 °C. Full article ">Figure 6 Ex vivo permeation profile of the pure drug and the complexed drug across the rat intestinal sac. Full article ">Figure 7 The plasma concentration–time curve of the pure drug (80 mg/kg p.o.) and complexed drug (equivalent to 80 mg/kg CBZ) administered in rats. Full article "> attachmentSupplementary material: Supplementary File 1 (ZIP, 2465 KiB)get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleComparison of the Purity and Impurity of Glucagon-for-Injection Products under Various Stability ConditionsbyZhongli Bao, Ya-Chi Cheng, Mary Ziping Luo and Jack Yongfeng Zhang Sci. Pharm. 2022, 90(2), 32; https://doi.org/10.3390/scipharm90020032 - 17 May 2022Abstract Glucagon is a polypeptide hormone that serves as an essential therapeutic agent in the emergency treatment of hypoglycemia. Recently, the first generic glucagon for injection was approved. However, unlike its brand name counterpart, which is produced via recombinant DNA, the generic glucagon is [...] Read more.Glucagon is a polypeptide hormone that serves as an essential therapeutic agent in the emergency treatment of hypoglycemia. Recently, the first generic glucagon for injection was approved. However, unlike its brand name counterpart, which is produced via recombinant DNA, the generic glucagon is produced using a chemical synthesis method. Regardless of its origin, impurities may occur in both glucagon drug products. While these impurities may greatly compromise the safety and efficacy of the glucagon drug products, studies accessing the impurities of glucagon for injection are limited. This manuscript analyzed the stability and impurities of a generic and brand glucagon for injection, including desamido and non-desamido impurities, under various storage and temperature conditions using an ultra-performance liquid chromatography method. The glucagon products were analyzed after 6 and 24 months of storage under room temperatures (20–25 °C). In addition, the products were also assessed after 6 months of storage under high temperatures (40 °C). Under each stability storage condition, three lots of the synthetic glucagon were evaluated by UPLC with at least one lot of the recombinant glucagon for comparison. A total of 37 peaks were identified (except for the solvent peaks, which appeared at retention times less than 1.5 min) from the synthetic and recombinant glucagon lots. It was found that the number of impurities observed in the synthetic glucagon were lower than the referenced recombinant glucagon across all stability conditions. Throughout all tested conditions, the synthetic glucagon for injection had an averaged purity of 92.8–99.3%, while the referenced recombinant drug had an averaged purity of 70.3–91.7%. Based on the study results, it can be concluded that the impurity profile for the synthetic glucagon for injection has a comparable and even lower level of impurities than the recombinant version under all stability conditions.Full article(This article belongs to the Special Issue Feature Papers in Scientia Pharmaceutica)►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 Amino acid structure of glucagon. Blue shaded sites are susceptible to deamidation modification. Full article ">Figure 2 G001 = AMP-glucagon; RLD = ELI-glucagon. (A). Representative chromatograms at zero time, 25 °C. (B). Representative chromatograms at 6 months, 25 °C. (C). Representative chromatograms at 24 months, 25 °C. (D). Representative chromatograms at 6 months, 40 °C. Full article ">Figure 2 Cont. G001 = AMP-glucagon; RLD = ELI-glucagon. (A). Representative chromatograms at zero time, 25 °C. (B). Representative chromatograms at 6 months, 25 °C. (C). Representative chromatograms at 24 months, 25 °C. (D). Representative chromatograms at 6 months, 40 °C. Full article ">Figure 2 Cont. G001 = AMP-glucagon; RLD = ELI-glucagon. (A). Representative chromatograms at zero time, 25 °C. (B). Representative chromatograms at 6 months, 25 °C. (C). Representative chromatograms at 24 months, 25 °C. (D). Representative chromatograms at 6 months, 40 °C. Full article "> get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleAntifungal and Modulatory Activity of Lemon Balm (Lippia alba (MILL.) N. E. BROWN) Essential OilbyGleilton Sales, Suelen Medeiros, Igor Soares, Tiago Sampaio, Mary Bandeira, Nádia Nogueira and Maria Queiroz Sci. Pharm. 2022, 90(2), 31; https://doi.org/10.3390/scipharm90020031 - 11 May 2022Abstract Fungal diseases and the progressive development of resistance are a challenge. In this context, Lippia alba (lemon balm) is a species used in folk medicine, being described with antimicrobial potential. The aim of this study was to determine the antifungal activity and modulating [...] Read more.Fungal diseases and the progressive development of resistance are a challenge. In this context, Lippia alba (lemon balm) is a species used in folk medicine, being described with antimicrobial potential. The aim of this study was to determine the antifungal activity and modulating effect of the essential oil of Lippia alba (Mill.) N.E. Brown leaves (LaEO). The antifungal activity of LaEO on eight Candida strains was determined by minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC), minimum biofilm inhibition concentration (MBIC), minimum biofilm eradication concentration (MBEC) and time-kill. The checkerboard technique was used to determine the modulating effect of LaEO on antifungal activity. The results indicate the presence of 11 constituents, with a predominance of carvone (58.15%) and limonene (25.37%). LaEO was able to inhibit the growth of all tested microorganisms, with MIC and MLC ranging from 0.078 to 1.25 mg/mL and MBIC and MBEC ranging from 0.156 to 5 mg/mL. The time-kill assay showed that LaEO was able to eliminate the strains after two hours of exposure and the best association was observed for the combination of LaEO and ketoconazole. The results of the study indicate that LaEO has excellent antifungal activity with potential biotechnological application.Full article►▼Show FiguresFigure 1Figure 1 Effect of the time of exposure on the minimum inhibitory concentration of LaEO on the viability of Candida strains, determined by the microdilution technique in culture broth and viable cell count. (A) MIC: 0.078 mg/mL. (B) MIC: 0.625 mg/mL. (C) MIC: 0.625 mg/mL. (D) MIC: 0.312 mg/mL. (E) MIC: 0.625 mg/mL. (F) MIC: 1.250 mg/mL. (G) MIC: 0.625 mg/mL. (H) MIC: 0.156 mg/mL. Control for all groups (C): culture medium + microorganism + 1% Tween 80. The values are expressed as mean ± SEM of three experiments, with three replicates. Data analysis was performed by ANOVA, with Tukey post-test and p < 0.05 (*). Full article "> attachmentSupplementary material:Supplementary File 1 (ZIP, 614 KiB)get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleCinnamomum bejolghota Extract Inhibits Colorectal Cancer Cell Metastasis and TGF-β1-Induced Epithelial-Mesenchymal Transition via Smad and Non-Smad Signaling PathwaybyAthicha Kittiwattanokhun, Sukanda Innajak, Etsu Tashiro, Masaya Imoto and Ramida Watanapokasin Sci. Pharm. 2022, 90(2), 30; https://doi.org/10.3390/scipharm90020030 - 09 May 2022Abstract Cinnamomum bejolghota, used in Thai traditional medicine remedies, has several biological activities including antimicrobial, antifungal, and anticancer. In colorectal cancer, epithelial-mesenchymal transition (EMT) is an initial step of cancer metastasis. Thus, this study investigated the effects of C. bejolghota bark extract (CBE) [...] Read more.Cinnamomum bejolghota, used in Thai traditional medicine remedies, has several biological activities including antimicrobial, antifungal, and anticancer. In colorectal cancer, epithelial-mesenchymal transition (EMT) is an initial step of cancer metastasis. Thus, this study investigated the effects of C. bejolghota bark extract (CBE) on colorectal cancer cell metastasis and transforming growth factor-β1 (TGF-β1) induced EMT in LoVo cells. The results showed that CBE could reduce cell migration, invasion, and adhesion of LoVo cells in a dose-dependent manner. In addition, our studies also showed that CBE could reverse TGF-β1-induced morphological changes as well as increase an epithelial marker, E-cadherin, while the expression of the mesenchymal marker, N-cadherin, was decreased in TGF-β1-treated LoVo cells. MMP-2 expression was effectively decreased but TIMP-1 and TIMP-2 expression was increased by the CBE treatment in LoVo cells. CBE also inhibited Smad2/3 phosphorylation and nuclear translocation as well as decreased the expression of Snail, Slug, and TCF8/ZEB1 transcription factors in LoVo cells. Moreover, CBE could inhibit TGF-β1-induced Smad-independent signaling pathway by decreased phosphorylation of ERK1/2, p38, and Akt. These findings suggest that CBE inhibited TGF-β1-induced EMT in LoVo cells via both Smad-dependent and Smad-independent pathways. Therefore, CBE may function as an alternative therapeutic treatment for colorectal cancer metastasis.Full article►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 Effect of CBE on cell viability in LoVo cells. Cells were treated with CBE for 24 and 48 h and cell viability was determined using MTT assay. Results are presented as mean values ± SD (n = 3). * p < 0.05 versus the control group. CBE, Cinnamomum bejolghota extract; SD, standard deviation. Full article ">Figure 2 Effect of CBE on migration, invasion, and adhesion of LoVo cells. (A) LoVo cells were treated with or without CBE at 0, 24, and 48 h and detected by scratch assay and wound area was analyzed by Image J. (B) Transwell migration assay after treatment of CBE for 24 h. (C) Transwell invasion assay after treatment of CBE for 24 h. (D) The adhesive ability of CBE-treated LoVo cells were determined by Matrigel-adhesion assay. Results are presented as mean values ± SD (n = 3). * p < 0.05 versus the control group. CBE, Cinnamomum bejolghota extract; SD, standard deviation. Full article ">Figure 2 Cont. Effect of CBE on migration, invasion, and adhesion of LoVo cells. (A) LoVo cells were treated with or without CBE at 0, 24, and 48 h and detected by scratch assay and wound area was analyzed by Image J. (B) Transwell migration assay after treatment of CBE for 24 h. (C) Transwell invasion assay after treatment of CBE for 24 h. (D) The adhesive ability of CBE-treated LoVo cells were determined by Matrigel-adhesion assay. Results are presented as mean values ± SD (n = 3). * p < 0.05 versus the control group. CBE, Cinnamomum bejolghota extract; SD, standard deviation. Full article ">Figure 3 Effect of CBE on reversion of TGF-β1-induced morphological changes in LoVo cells. Cells were pre-treated with CBE and SB431542. After incubation, cells were treated with TGF-β1 for 24 and 48 h. Cell morphology was observed under a microscope. CBE, Cinnamomum bejolghota extract. Full article ">Figure 4 Effect of CBE on TGF-β1-induced EMT-related biomarker proteins expression. Cells were pre-treated with CBE and SB431542 then stimulated with TGF-β1 for an additional 24 and 48 h. (A) Western blotting analysis of E-cadherin and N-cadherin EMT marker protein. (B) qPCR analysis of N-cadherin mRNA expression. (C) Western blotting analysis of MMP-2, TIMP-1, and TIMP-2. β-actin was used as an internal control. The amount of N-cadherin mRNA was normalized using RPL37A mRNA as a reference. Results are presented as mean values ± SD (n = 3). # p < 0.05 versus the control group. * p < 0.05 versus the TGF-β1 treated group. CBE, Cinnamomum bejolghota extract; SD, standard deviation. Full article ">Figure 4 Cont. Effect of CBE on TGF-β1-induced EMT-related biomarker proteins expression. Cells were pre-treated with CBE and SB431542 then stimulated with TGF-β1 for an additional 24 and 48 h. (A) Western blotting analysis of E-cadherin and N-cadherin EMT marker protein. (B) qPCR analysis of N-cadherin mRNA expression. (C) Western blotting analysis of MMP-2, TIMP-1, and TIMP-2. β-actin was used as an internal control. The amount of N-cadherin mRNA was normalized using RPL37A mRNA as a reference. Results are presented as mean values ± SD (n = 3). # p < 0.05 versus the control group. * p < 0.05 versus the TGF-β1 treated group. CBE, Cinnamomum bejolghota extract; SD, standard deviation. Full article ">Figure 5 CBE reduces TGF-β1-induced EMT-regulated transcription factors. Cells were pre-treated with CBE and SB431542 then stimulated with TGF-β1 for an additional 24 and 48 h. (A) Western blot analysis of transcription factors Snail, Slug, and TCF8/ZEB1 were detected. β-actin was used as an internal control. (B) qPCR analysis of Snail and Slug mRNA expression were detected after TGF-β1 stimulation for 24 h, the amount of Snail, Slug, and RPL37A mRNA were quantified and normalized using RPL37A mRNA as a reference. Results are presented as mean values ± SD (n = 3). # p < 0.05 versus the control group. * p < 0.05 versus the TGF-β1-treated group. CBE, Cinnamomum bejolghota extract; SD, standard deviation. Full article ">Figure 6 CBE reduces TGF-β1-induced Smad2/3 phosphorylation. LoVo cells were pre-treated with CBE and SB431542, and were then stimulated with TGF-β1 for an additional 1 h. TGFβR-I, Smad2/3, and p-Smad2/3 was determined by Western blotting. Results are presented as mean values ± SD (n = 3). # p < 0.05 versus the control group. * p < 0.05 versus the TGF-β1 treated group. CBE, Cinnamomum bejolghota extract; SD, standard deviation. Full article ">Figure 7 Effect of CBE on Smad2/3 nuclear translocation. Cells were pre-treated with CBE and SB431542, then stimulated with TGF-β1 for an additional 1 h. (A) Smad2/3 nuclear translocation was monitored by immunofluorescence analysis. (B) Smad2/3 localization was observed by cytoplasmic/nuclear fractionation assay. β-Tubulin and PARP were used as a control for the cytoplasmic and nuclear fraction, respectively. Data are mean values ± SD (n = 3). # p < 0.05 versus the TGF-β1-treated group. * p < 0.05 compared to the TGF-β1-treated group. CBE, Cinnamomum bejolghota extract; SD, standard deviation. Full article ">Figure 8 Effect of CBE on TGF-β1-induced EMT via Smad-independent signaling pathway in LoVo cells. Cells were pre-treated with 10, 30 and 100 µg/mL of CBE and 1 µM SB431542. After incubation, cells were treated with 1 ng/mL TGF-β1 for an additional 24 and 48 h. (A) Western blot analysis of p-ERK1/2, ERK1/2, p-p38, and p38 the proteins of MAPK signaling pathway. (B) Western blot analysis of p-Akt (Ser473) and Akt, proteins of Akt signaling pathway. β-actin was used as an internal control. Data are mean values ± SD of three independent experiments (n = 3). # p < 0.05 compared to the control group. * p < 0.05 compared to the TGF-β1-treated group. CBE, Cinnamomum bejolghota extract; SD, standard deviation. Full article ">Figure 9 Effect of cinnamaldehyde on TGF-β1- induced Smad2/3 phosphorylation and EMT markers in LoVo cells. (A) Cell viability was determined using MTT assay. (B) Cells were pre-treated with cinamaldehyde for 30 min, then cultured in TGF-β1 for an additional 24 and 48 before detection of morphological changes. (C) The phosphorylation of Smad2/3 was determined by Western blot analysis. (D) Western blotting of the E-cadherin and N-cadherin EMT marker proteins. β-actin was used as an internal control. Data are mean values ± SD of three independent experiments (n = 3). # p < 0.05 compared to the control group. * p < 0.05 compared to the TGF-β1 treated group. CBE, Cinnamomum bejolghota extract; SD, standard deviation. Full article ">Figure 9 Cont. Effect of cinnamaldehyde on TGF-β1- induced Smad2/3 phosphorylation and EMT markers in LoVo cells. (A) Cell viability was determined using MTT assay. (B) Cells were pre-treated with cinamaldehyde for 30 min, then cultured in TGF-β1 for an additional 24 and 48 before detection of morphological changes. (C) The phosphorylation of Smad2/3 was determined by Western blot analysis. (D) Western blotting of the E-cadherin and N-cadherin EMT marker proteins. β-actin was used as an internal control. Data are mean values ± SD of three independent experiments (n = 3). # p < 0.05 compared to the control group. * p < 0.05 compared to the TGF-β1 treated group. CBE, Cinnamomum bejolghota extract; SD, standard deviation. Full article "> get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleGossypol from Gossypium spp. Inhibits Helicobacter pylori Clinical Strains and Urease Enzyme Activity: Bioactivity and Safety AssessmentsbyMiroslava Šudomová and Sherif T. S. Hassan Sci. Pharm. 2022, 90(2), 29; https://doi.org/10.3390/scipharm90020029 - 05 May 2022Abstract This study investigates the inhibitory activities of gossypol, a natural polyphenolic compound from Gossypium spp., against Helicobacter pylori (HP) clinical strains and a urease enzyme that plays a key role in the pathogenesis of HP. Gossypol was detected to exhibit a bacteriostatic action [...] Read more.This study investigates the inhibitory activities of gossypol, a natural polyphenolic compound from Gossypium spp., against Helicobacter pylori (HP) clinical strains and a urease enzyme that plays a key role in the pathogenesis of HP. Gossypol was detected to exhibit a bacteriostatic action against all the HP strains tested with minimum inhibitory concentration (MIC) values ranging from 3.51 to 4.14 µg/mL. The activity of HP urease (HPU) was efficiently impeded by gossypol with a 50% inhibitory concentration (IC50) value of 3.3 µM using an Electrospray Ionization–Mass Spectrometry (ESI-MS)-based method. The in vitro cytotoxicity assay showed no significant cytotoxic properties of gossypol against human gastric epithelial cells. Additionally, molecular docking studies were performed to assess the binding mode and the molecular interactions of gossypol with HPU with a binding affinity value of −8.1 kcal/mol compared with an HPU–acetohydroxamic acid (a standard urease inhibitor) docking complex (–6.1 kcal/mol). The overall results reveal that gossypol might help fight against HP infection by two mechanisms of action: inhibition of the growth of HP and inhibition of urease.Full article(This article belongs to the Special Issue Feature Papers in Scientia Pharmaceutica)►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 Chemical structure of gossypol. Full article ">Figure 2 The anti-Helicobacter pylori urease (HPU) activities of gossypol and acetohydroxamic acid (standard inhibitor) were assayed by an Electrospray Ionization–Mass Spectrometry (ESI-MS)-based method. The reaction rate constant (RRC) is shown on each slope. As displayed, k0 represents the RRC of the HPU-catalyzed reaction in the absence of inhibitors (k0 = 0.1081/min) (A) and k represents the RRC of the HPU-catalyzed reaction inhibited by gossypol (k = 0.0184/min) (B) and acetohydroxamic acid (k = 0.0244/min) (C). Changes in urea concentrations are expressed as logarithms of concentration. The precision of the time-course analysis was confirmed by the relative standard deviation (RSD; %) of multiple measured slopes (less than 10%). For figure clarity, multiple measurements are not presented. Gossypol and acetohydroxamic acid effectively inhibited HPU with 50% inhibitory concentration (IC50) values of 3.3 and 4.7 µM, respectively. Full article ">Figure 3 Molecular docking analysis (in a two-dimensional model) indicates the binding mode and the molecular interaction of gossypol with the Helicobacter pylori urease (HPU) active site. As shown, numerous crucial interactions between the functional groups of gossypol and amino acid residues of the active site of HPU are formed. Full article "> get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticlePreclinical Safety Profile of an Oral Naringenin/Hesperidin Dosage Form by In Vivo Toxicological TestsbyCarla Georgina Cicero-Sarmiento, Rolffy Ortiz-Andrade, Jesús Alfredo Araujo-León, Maira Rubí Segura-Campos, Priscila Vazquez-Garcia, Héctor Rubio-Zapata, Efrén Hernández-Baltazar, Victor Yañez-Pérez, Amanda Sánchez-Recillas, Juan Carlos Sánchez-Salgado, Emanuel Hernández-Núñez and Durcy Ruiz-Ciau Sci. Pharm. 2022, 90(2), 28; https://doi.org/10.3390/scipharm90020028 - 02 May 2022Abstract We developed a naringenin–hesperidin molar mixture (MIX–160) with proven antihyperglycemic and vasorelaxant activity in preclinical studies. A solid dosage form was manufactured to improve the bioavailability properties. In the current study, we sought to evaluate the oral preclinical toxicity of the MIX–160 dosage [...] Read more.We developed a naringenin–hesperidin molar mixture (MIX–160) with proven antihyperglycemic and vasorelaxant activity in preclinical studies. A solid dosage form was manufactured to improve the bioavailability properties. In the current study, we sought to evaluate the oral preclinical toxicity of the MIX–160 dosage form, which showed no mortality or significant changes in the body weight, food consumption and tissue/organ mass in rats. Three daily oral doses (50, 300 and 2000 mg/kg of MIX–160) were assayed for 28 days. The results showed no structural abnormalities in the histological analysis and no significant changes (p > 0.05) in the liver biochemical markers (total bilirubin, AST and ALT) compared to the control group. The above findings showed that the MIX–160 dosage form did not exhibit relevant toxic effects, which suggests its potential safety as a drug candidate for clinical studies.Full article(This article belongs to the Special Issue Feature Papers in Scientia Pharmaceutica)►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 Effects of MIX–160 on liver enzyme content (A) and bilirubin (B) in acute oral toxicity evaluations. ALT is represented in white with gray lines. AST is represented in solid gray. Full article ">Figure 2 Effects of MIX–160 on the (A) body weight, (B) tissue–mass percentage, (C) food consumption, (D) water consumption, (E) excreted feces and (F) urine in repeated dose 28–day oral toxicity evaluations. The control group is represented in light gray or white, and MIX–160 is represented in dark gray. Full article ">Figure 3 Effects of MIX–160 on (A) blood cells, (B) bilirubin and (C) AST and ALT transaminases in repeated dose 28–day oral toxicity evaluations. The control group is represented in white, and MIX–160 is represented in gray. Full article ">Figure 4 Histological evaluation of kidney, liver, and intestine tissues of the control vs. MIX–160 treated groups. Full article "> get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticlePurification of Andrographolide Methanolic Extract Using Molecularly Imprinted Polymer Prepared by Precipitation PolymerizationbyWiwin Winingsih, Slamet Ibrahim and Sophi Damayanti Sci. Pharm. 2022, 90(2), 27; https://doi.org/10.3390/scipharm90020027 - 26 Apr 2022Abstract Molecularly Imprinted Polymer (MIP) has a specific cavity in which the conformity of shape, size, and functionalities corresponds with its template molecule and has been widely used in separation processes. Therefore, this study aims to examine the application of MIP for the purification [...] Read more.Molecularly Imprinted Polymer (MIP) has a specific cavity in which the conformity of shape, size, and functionalities corresponds with its template molecule and has been widely used in separation processes. Therefore, this study aims to examine the application of MIP for the purification of andrographolide. The MIP was synthesized by precipitation polymerization using methacrylic acid (MAA) and ethylene glycol dimethacrylate (EGDMA) as the functional monomer and cross-linker, andrographolide as a template, and acetonitrile:toluene (3:1) as porogen solvent. The results showed that the binding capacity of Synthesized MIP was 1.2486 mg/g, while the particle size was 295.5 nm with a polydispersity index of 0.064. Furthermore, the imprinting and selectivity factors were 1.148 and 12.37, respectively. The purification process by MIP increased the purity from 55.37 ± 0.69 to 94.94% ± 0.34, while the isolate characterization showed that purified andrographolide had a similar character compared to the standard.Full article►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 Stucture of Andrographolide. Full article ">Figure 2 Predicted interaction between andrographolide and MAA (performed by Gaussian W 09). Full article ">Figure 3 Proposed scheme of reaction of MIP synthesis; a: extraction of template molecule; b: rebinding of template molecule. Full article ">Figure 4 Surface morphology of MIP (a) and NIP (b). Full article ">Figure 5 The size distribution curve of MIP (a) and NIP (b). Full article ">Figure 6 FTIR spectrum of MIP and NIP compared to EGDMA (cross-linker) and MAA (functional monomer). Full article ">Figure 7 The adsorption capacity of MIP and NIP after undergoing five cycles isotherm adsorption studies. Full article ">Figure 8 FTIR spectrum of MIP (a) and NIP (b) before and after undergo five cycles isotherm adsorption study. Full article ">Figure 9 Organoleptic appearance of andrographolide extract (a), isolate after purification (b), and standard (c). Full article ">Figure 10 FTIR spectrum of andrographolide standard (blue) and isolate (red). Full article ">Figure 11 Chromatogram of andrographolide standard (green), isolate (blue), and andrographolide extract (red). Full article ">Figure 12 Spectrum UV-Vis of andrographolide extract (purple), isolate (grey), and standard (black). Full article "> attachmentSupplementary material:Supplementary File 1 (ZIP, 3327 KiB)get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleBinding of GS-461203 and Its Halogen Derivatives to HCV Genotype 2a RNA Polymerase Drug Resistance MutantsbyMuhammad Arba, Setyanto Tri Wahyudi, Muhammad Sulaiman Zubair, Dylan Brunt, Mursalin Singh and Chun Wu Sci. Pharm. 2022, 90(2), 26; https://doi.org/10.3390/scipharm90020026 - 21 Apr 2022Abstract Hepatitis C Virus (HCV) is reported to develop GS-461203 resistance because of multiple mutations within the RNA-dependent RNA Polymerase (RdRp) of HCV. The lack of a high-resolution structure of these RdRp mutants in complex with GS-461203 hinders efforts to understand the drug resistance. [...] Read more.Hepatitis C Virus (HCV) is reported to develop GS-461203 resistance because of multiple mutations within the RNA-dependent RNA Polymerase (RdRp) of HCV. The lack of a high-resolution structure of these RdRp mutants in complex with GS-461203 hinders efforts to understand the drug resistance. Here we decipher the binding differences of GS-461203 in the wild type and mutated systems T179A or M289L of HCV RdRp Genotype 2a using homology modeling, molecular docking, and molecular dynamics simulation. Key residues responsible for GS-461203 binding were identified to be Arg48, Arg158, Asp318, Asp319, and Asp220, and that mutations T179A or M289L have caused conformational changes of GS-461203 in the RdRp active site. The affinities of GS-461203 were reduced in T179A system, but it became slightly stronger in the M289L system. Furthermore, we designed two new analogues of GS-461203 which encouragingly induced more stable interactions than GS-461203, and thus resulted in much better binding energies. This present study reveals how a single mutation, T179A or M289L, will modulate GS-461203 binding in HCV RdRp Genotype 2a, while introducing two novel analogues to overcome the drug resistance which may be good candidate for further experimental verification.Full article►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 The docked poses of GS−461203 (A) in WT (green), T179A (blue) and M289L (purple), two manganese ions (purple), with the positions of mutated residues; and the chemical structure of the drug (B). Full article ">Figure 2 The root mean squared deviation (RMSD) of Protein Cα and ligand averaged over the two runs in (A) WT, (B) T179A, S282T, and (C) M289L systems. Full article ">Figure 3 The 2D Protein–Ligand Interaction Diagrams lasting more than 20% of the MDS (A) WT-GS-461203; (B) T179A-GS-461203; (C) M289L-GS-461203 recorded during MD simulations. Full article ">Figure 3 Cont. The 2D Protein–Ligand Interaction Diagrams lasting more than 20% of the MDS (A) WT-GS-461203; (B) T179A-GS-461203; (C) M289L-GS-461203 recorded during MD simulations. Full article ">Figure 4 Comparison of protein structure between WT (green) and T179A (red) (A) WT vs. T179A, RMSD = 2.76 Å and M289L (red) (B) WT vs. M289L, RMSD = 3.95 Å. Full article ">Figure 5 The active site conformation of WT (A), T179A (B), and M289L (C) systems taken from most populated cluster. Full article ">Figure 6 The representative structure of the most dominant structural family for (A) WT; (B) T179A; (C) M289L systems. Full article ">Figure 7 The RMSF values comparison between WT and T179A (A) and WT and M289L (B). The RMSF values of the Cα atoms for residues in WT (black), T179A (green), and M289L (blue) are shown with the mutant positions are noted by yellow asterisks. Full article ">Figure 8 The RMSF values for atoms of GS-461203. Full article ">Figure 9 The dihedral angle of GS−461203 profiles during 500 ns MDS which is the conformational progression of the nine rotatable bonds of GS-461203. The dial plots describe the conformation of the torsion throughout the course of the simulation. The beginning of the simulation is in the center of the radial plot and the time evolution is plotted radially outwards. The bar plots summarize the data on the dial plots, by showing the probability density of the torsion. In addition, the 2D structure of GS-461203 in the top panel is for reference. Full article ">Figure 10 Protein SSE for the WT, T179A, and M289L systems during MDS. The alpha helices, beta sheets, and random coil were represented by red, blue, and white spaces. Full article ">Figure 11 The designed GS−461203 analogues. Full article "> get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleCompounding in Ukraine: Assessment of the Risks for the Ointment’s Quality by the FMECA MethodbyLesia Savchenko, Yuri Pidpruzhnykov, Roman Lesyk, Liudas Ivanauskas, Alla Kotvitska and Victoriya Georgiyants Sci. Pharm. 2022, 90(2), 25; https://doi.org/10.3390/scipharm90020025 - 19 Apr 2022Abstract The level of compounded medicines (CM) quality has always been questioned in different countries. This problem has been resolved by the introduction of quality assurance system (QAS) standards. One of its main areas of significance is the risks assessment process, which is especially [...] Read more.The level of compounded medicines (CM) quality has always been questioned in different countries. This problem has been resolved by the introduction of quality assurance system (QAS) standards. One of its main areas of significance is the risks assessment process, which is especially important for the compounding pharmacy according to the requirements of different international documents. Since ointments constitute a large part of CM, quantity assessment of risks for their quality by the FMECA method has been completed. During the first step of the research, 42 potential deviations of compounded ointments (CO) quality were identified. Via the questioning of compounding pharmacies specialists in different regions of Ukraine by a pre-developed ten-point scale, the severity of deviations consequence, their occurrence probability, and detecting possibility were determined followed by the calculation of the priority risk number (PRN) value. The Pareto analysis showed that nine possible CO quality defects represented 21% of their total number. Defects related to the composition or technology of ointments (29%) and their compliance with microbiological purity requirements (23%) had the largest percentage contribution to the total PRN value. It was also found that the deviations consequence had the most serious impact on the CO quality, due to their direct influence on patient health.Full article(This article belongs to the Special Issue Feature Papers in Scientia Pharmaceutica)►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 Research design. Full article ">Figure 2 Main elements of the QAS in compounding pharmacies. Full article ">Figure 3 Possible deviations in compounded ointments quality. Full article ">Figure 4 Percentage of each region’s participation in the survey (by the pharmacies quantity). Full article ">Figure 5 The value of the average score of defects in pharmacies. Notes: 1—Kirovograd region; 2—Khmelnytskyi region; 3—Dnipropetrovsk region; 4—Sumy region; 5—Odesa region; 6–12—Mykolaiv region; 13–15—Kharkiv region; 16–18—Luhansk region; 19–21—Zhytomyr region; 22–28—Rivne region. Full article ">Figure 6 PRN average value analysis by the Pareto method. Full article "> attachmentSupplementary material:Supplementary File 1 (ZIP, 1704 KiB)get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleRepurposing of Four Drugs as Anti-SARS-CoV-2 Agents and Their Interactions with Protein TargetsbyLuis C. Vesga, Camilo A. Ruiz-Hernández, Jeimmy J. Alvarez-Jacome, Jonny E. Duque, Bladimiro Rincon-Orozco and Stelia C. Mendez-Sanchez Sci. Pharm. 2022, 90(2), 24; https://doi.org/10.3390/scipharm90020024 - 14 Apr 2022Abstract Although there are existing vaccines against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), new COVID-19 cases are increasing due to low immunization coverage and the emergence of new variants. For this reason, new drugs to treat and prevent severe COVID-19 are needed. Here, we [...] Read more.Although there are existing vaccines against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), new COVID-19 cases are increasing due to low immunization coverage and the emergence of new variants. For this reason, new drugs to treat and prevent severe COVID-19 are needed. Here, we provide four different FDA-approved drugs against SARS-CoV-2 proteins involved in the entry and replication process, aiming to identify potential drugs to treat COVID-19. We use the main protease (Mpro), the spike glycoprotein (S protein), and RNA-dependent RNA polymerase (RdRp) as protein targets for anti- SARS-CoV-2 drugs. In our constructed database, we selected different drugs against each target (Mpro, S protein, and RdRp) based on their common interactions with relevant residues involved in viral entry at the host cell and replication. Furthermore, their stability inside the binding pocket, as well as their predicted binding-free energy, allow us to provide new insight into the possible drug repurposing of viomycin (interacting with Mpro) due to its interactions with key residues, such as Asn 143, Glu 166, and Gln 189 at the same time as hesperidin (interacting with the S protein) is interacting with residues Tyr 449, Ser 494, and Thr 500, keeping inside the predicted binding pocket, as well as interacting with residues in different variants of concern. Finally, we also suggest nystatin and elvitegravir (interacting with RdRp) as possible drugs due to their stability within the predicted pocket along the simulation and their interaction with key residues, such as Asp 760, Asp 761, and Asp 618. Altogether our results provide new knowledge about the possible mechanism of the inhibition of viomycin, hesperidin, elvitegravir, and nystatin to inhibit the viral life cycle of SARS-CoV-2 and some of its variants of concern (VOC). Additionally, some iodide-based contrast agents were also found to bind the S protein strongly, i.e., iohexol (−58.99 Kcal/mol), iotrolan (−76.19 Kcal/mol), and ioxilan (−62.37 Kcal/mol). Despite the information we report here as the possible strong interaction between these contrast agents and the SARS-CoV-2′s S protein, Mpro, and RdRp, we believe that further investigation, including chemical modifications in their structures, are needed for COVID-19 treatment.Full article(This article belongs to the Special Issue Feature Papers in Scientia Pharmaceutica)►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 A representative snapshot of the docking pose of viomycin. Mpro’s residues are colored according to the atom type of the interacting amino acid residues (protein’s carbon, light grey; oxygen, red; nitrogen, blue). The protein–ligand interactions are represented by dash lines as follows: hydrogen bond interactions are colored in yellow. Full article ">Figure 2 Viomycin protein–ligand interaction along the 100 ns simulation. Frequency of hydrogen bonding interactions (A); water-mediated hydrogen bonding interactions (B); hydrophobic interactions (C); representative picture of root mean square deviation (RMSD) values of protein backbone for the protein–viomycin complexes (D); ligand RMSD variation along the simulation time for viomycin (E). Full article ">Figure 3 The interface between the viral S protein and ACE2 host cell receptor. Residues are colored according to the atom type of the interacting amino acid residues (the S protein’s carbon, pale green; the ACE2 receptor’s carbon, red; oxygen, red; nitrogen, blue). Dash lines represent the protein–ligand interactions: hydrogen bond interactions are colored in yellow. Figure adapted from Unni et al., 2020. Full article ">Figure 4 A representative snapshot of the docking pose of hesperidin. The S protein’s residues are colored according to the atom type of the interacting amino acid residues (protein’s carbon, pale green; oxygen, red; nitrogen, blue). Dash lines represent the protein–ligand interactions: hydrogen bond interactions are colored in yellow; π-π interactions are colored in blue. Full article ">Figure 5 Hesperidin protein–ligand interaction along the 100 ns simulation. Frequency of hydrogen bonding interactions (A); water-mediated hydrogen bonding interactions (B); hydrophobic interactions (C); π-π interactions (D); representative picture of root mean square deviation (RMSD) values of protein backbone for the protein–hesperidin complexes (E); Ligand RMSD variation along the simulation time for viomycin (F). Full article ">Figure 6 A representative snapshot of the docking pose of hesperidin with the Alpha variant. B.1.1.7 (Alpha’s) residues are colored according to the atom type of the interacting amino acid residues (protein’s carbon, pale green; oxygen, red; nitrogen, blue). Dash lines represent the protein–ligand interactions: hydrogen bond interactions are colored in yellow; π-π interactions are colored in blue. Full article ">Figure 7 A representative snapshot of the docking pose of hesperidin with the Beta variant. B.1.351 (Beta’s) residues are colored according to the atom type of the interacting amino acid residues (protein’s carbon, pale green; oxygen, red; nitrogen, blue). Dash lines represent the protein–ligand interactions: hydrogen bond interactions are colored in yellow; π-π interactions are colored in blue; π-cation interactions are colored in green. Full article ">Figure 8 A representative snapshot of the docking pose of hesperidin with the Delta variant. B.1.617.2 (Delta’s) residues are colored according to the atom type of the interacting amino acid residues (protein’s carbon, pale green; oxygen, red; nitrogen, blue). Dash lines represent the protein–ligand interactions: hydrogen bond interactions are colored in yellow; π-π interactions are colored in blue; π-cation interactions are colored in green. Full article ">Figure 9 Nystatin and elvitegravir protein–ligand interaction along the 100 ns simulation. Frequency of hydrogen bonding interactions (A); hydrophobic interactions (B); water-mediated hydrogen bonding interactions (C); ionic interactions (D). Full article ">Figure 10 Nystatin-selected docking pose. RdRp’s residues are colored according to the atom type of the interacting amino acid residues (protein’s carbon, light teal; oxygen, red; nitrogen, blue). Dash lines represent the protein–ligand interactions: hydrogen bond interactions are colored in yellow; π-π interactions are colored in blue. Full article ">Figure 11 Elvitegravir-selected docking pose. RdRp’s residues are colored according to the atom type of the interacting amino acid residues (protein’s carbon, light teal; oxygen, red; nitrogen, blue). Dash lines represent the protein–ligand interactions: hydrogen bond interactions are colored in yellow; π-π interactions are colored in blue; π-cation interactions are colored in green. Full article "> get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleEnhanced Antibacterial Activity of Brevibacillus sp. SPR19 by Atmospheric and Room Temperature Plasma Mutagenesis (ARTP)byNuttapon Songnaka, Mudtorlep Nisoa, Apichart Atipairin, Thamonwan Wanganuttara and Thapanee Chinnawong Sci. Pharm. 2022, 90(2), 23; https://doi.org/10.3390/scipharm90020023 - 06 Apr 2022Abstract Antibiotic resistance is a major health concern worldwide. In our previous study, some bacterial isolates exhibited antibacterial activity against Staphylococcus aureus and methicillin-resistant S. aureus (MRSA). However, the production of antibacterial substances by native microorganisms is limited by biosynthetic genes. This study [...] Read more.Antibiotic resistance is a major health concern worldwide. In our previous study, some bacterial isolates exhibited antibacterial activity against Staphylococcus aureus and methicillin-resistant S. aureus (MRSA). However, the production of antibacterial substances by native microorganisms is limited by biosynthetic genes. This study aimed to improve the antibacterial activity of SPR19 using atmospheric and room temperature plasma mutagenesis (ARTP). The results showed that SPR19 belonged to the Brevibacillus genus. The growth curves and production kinetics of antibacterial substances were investigated. Argon-based ARTP was applied to SPR19, and the 469 mutants were preliminarily screened using agar overlay method. The remaining 25 mutants were confirmed by agar well diffusion assay against S. aureus TISTR 517 and MRSA isolates 142, 1096, and 2468. M285 exhibited the highest activity compared to the wild-type strain (10.34–13.59%) and this mutant was stable to produce the active substances throughout 15 generations consistently. The antibacterial substances from M285 were tolerant to various conditions (heat, enzyme, surfactant, and pH) while retaining more than 90% of their activities. Therefore, Brevibacillus sp. SPR19 is a potential source of antibacterial substances. ARTP mutagenesis is a powerful method for strain improvement that can be utilized to treat MRSA infection in the future.Full article►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 Phylogenetic tree of Brevibacillus sp. SPR19 based on 16s rRNA sequencing. The tree was constructed by MEGA X software with neighbor-joining analysis. Full article ">Figure 2 Growth curve of Brevibacillus sp. SPR19 and its production kinetics of antibacterial substances. The growth curve was measured at OD 600 nm, while the antibacterial activity against S. aureus TISTR 517 and MRSA isolates 142, 1096, and 2468 was determined by the agar well diffusion method. The experiment was performed in triplicate, and the results were expressed as mean ± SD (n = 3). Full article ">Figure 3 The lethality rate of Brevibacillus sp. SPR19 by ARTP treatment. The experiment was performed in triplicate, and the results were expressed as mean ± SD (n = 3). Full article ">Figure 4 (a) The hydrogen peroxide, (b) nitrite concentration, and (c) pH after ARTP treatment. The experiments were performed in triplicate, and the results were expressed as mean ± SD (n = 3). Full article ">Figure 5 Antibacterial activity of the wild-type and its mutants (SPR19) after 15 consecutive generations. The CFS of the wild-type and mutants were used to measure the antibacterial activity against (a) S. aureus TISTR 517, (b) MRSA isolate 142, (c) MRSA isolate 1096, and (d) MRSA isolate 2468. The experiments were performed in triplicate, and the results were expressed as mean ± SD (n = 3). Full article ">Figure 6 Colony morphology of Brevibacillus sp. SPR19 of (a) wild-type and (b) M285 strains. The SEM micrograph of vegetative cells and spores of (c) wild-type and (d) M285 strains at magnitude 10,000×. The vegetative cells and spores were obtained by cell incubation on MH agar at 30 °C for 8 d. Full article ">Figure 7 Growth curve and production kinetics of antibacterial substances of the wild-type and M285 strains. Full article "> get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleCinnamaldehyde Relieves Induced Hepatocellular Carcinoma in Rat Model via Targeting Wnt/β-Catenin PathwaybyAl Shaima G. Abd El Salam, Yara A. Samra and Mamdouh M. El-Shishtawy Sci. Pharm. 2022, 90(2), 22; https://doi.org/10.3390/scipharm90020022 - 31 Mar 2022Abstract Cinnamaldehyde (CA) is a natural compound that has promising biological activity. The current study investigates the antitumor activity of CA in thioacetamide induced hepatocellular carcinoma (HCC) in rats through targeting the Wnt/β-catenin pathway and evaluates the capability of CA to relieve hepatocytes oxidative [...] Read more.Cinnamaldehyde (CA) is a natural compound that has promising biological activity. The current study investigates the antitumor activity of CA in thioacetamide induced hepatocellular carcinoma (HCC) in rats through targeting the Wnt/β-catenin pathway and evaluates the capability of CA to relieve hepatocytes oxidative stress in the HCC-rat model. After 16 weeks of HCC induction by thioacetamide (TAA), rats were treated for 7 consecutive weeks with CA daily; i.p. injection, Alpha-fetoprotein (AFP) level, necroinflammatory score and fibrosis percentage were measured to assess HCC development. The Wnt/β-catenin pathway was evaluated by measuring the hepatic protein level of Wnt-3a, β-catenin, cyclin D, matrix metalloproteinase-9 (MMP-9), and vascular endothelial growth factor (VEGF). Furthermore, hepatocytes’ oxidative stress was assessed by measuring hepatic GSH and MDA contents. Results showed that CA was significantly inhibiting the Wnt/β-catenin pathway through the downregulation of hepatic Wnt-3a, β-catenin, cyclin D, MMP-9, and VEGF. Moreover, CA ameliorates hepatocytes’ oxidative stress via lowering hepatic MDA content and rising hepatic GSH content. Thus, in conclusion, CA is a promising treatment for HCC. It not only has an effective antitumor activity but also ameliorates hepatocytes’ oxidative stress.Full article►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 CA decreases in fibrosis percentage (A) and necroinflammatory score (B) in liver homogenate. Data in A were expressed as mean ± S.E.M and data in B were expressed as median and range. Ten rats in each group. ++ p < 0.01, +++ p< 0.001 vs. control group, ### p < 0.001 vs. HCC group. Full article ">Figure 2 Effect of CA on serum AFP level. Data were expressed as mean ± S.E.M. Ten rats in each group. +++ p < 0.001 vs. control group, ### p < 0.001 vs. HCC group. Full article ">Figure 3 Histopathological examination of liver tissue in control, HCC and CA groups. H&E-stained liver sections. Hepatic sections show the normal arrangement of hepatic cords around central vein (CV) with the normal portal areas and sinusoids in control group. In contrast, hepatic sections from HCC group showing disrupted parenchymal structure due to extensive fibrosis (black arrows), scored six, associated with inflammatory cells infiltration that includes hemosiderin laden macrophages, congested blood vessels (red arrows). Hepatocytes are present in solid nodules (yellow arrows), lacking normal structures of hepatic lobules, suffered from microvascular degeneration (arrowheads) to ballooning degeneration (blue arrows) and necrosis (green arrows). Hepatic sections from CA group showing slightly improved hepatic parenchymal structure characterized by portal fibrosis (black arrows), scored four, associated with inflammatory cells infiltration (dashed arrows). Hepatocytes suffered from macrovascular degeneration (arrowheads) to hydropic degeneration (thick arrows). Ten rats in each group. Scale bar = 100 µm (A,C–E,G,H), Scale bar = 50 µm (B,F,I,J). Full article ">Figure 4 Histopathological examination of liver tissue in control, HCC and CA groups. Masson’s trichome stained liver sections. Microscopic pictures of hepatic sections from control group showing no collagen deposition. Meanwhile, hepatic sections from HCC group showed excessive bluish collagen deposition (black arrows) dividing hepatic lobules into separate nodules. Hepatic sections from CA group showed mild perivascular bluish collagen deposition (black arrows) with thin collagen strands extending from portal areas (red arrow). Ten rats in each group. Scale bar = 100 µm (A,C,E), Scale bar = 50 µm (B,D,F). Full article ">Figure 5 Effect of CA on hepatic (A) Wnt-3a, (B) β-catenin, (C) Cyclin D, and (D) MMP-9 protein levels. Data were expressed as mean ± S.E.M. Ten rats in each group. +++ p < 0.001 vs. control group, ### p < 0.001 vs. HCC group. Full article ">Figure 6 Effect of CA on VEGF expression within the hepatic tissue. (A) Immunohistochemical stained liver sections of VEGF, black arrows indicate VEGF antibody positive regions. Scale bar = 50 µm. (B) VEGF percentage of positive cells in hepatic tissue. Data were expressed as mean ± S.E.M. Te rats in each group. +++ p < 0.001 vs. control group, ### p < 0.001 vs. HCC group. Full article ">Figure 7 Immunohistochemical stained liver sections of β-catenin. Control group (A,B); HCC group (C,D); CA group (E,F). Black arrowheads indicate nuclear localization of β-catenin positive cells and black arrows indicate cytoplasmic localization of β-catenin positive cells. X 200 (A,C,E), X 400 (B,D,F). Full article ">Figure 8 Effect of CA on TAA induced hepatic oxidative stress parameters. (A) Hepatic MDA content and (B) Hepatic GSH content. Data were expressed as mean ± S.E.M. Ten rats in each group. +++ p < 0.001 vs. control group, ### p < 0.001 vs. HCC group. Full article "> get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleTemperature and pH-Dependent Behaviors of mAb Drugs: A Case Study for TrastuzumabbyFatma Sert, Defne Hız, Mert Gülmez, Selen Ezgi Cankurtaran, Cemre Irmak Kayalan, Hasan Kurt and Meral Yüce Sci. Pharm. 2022, 90(1), 21; https://doi.org/10.3390/scipharm90010021 - 21 Mar 2022Cited by 1Abstract The distortions in the high-order structure of therapeutic monoclonal antibodies (mAbs) under different environmental conditions acutely affect mAb stability, resulting in altered safety, efficacy, and shelf-life profiles. The overall stability of mAbs depends on many factors, and it requires complementary techniques for an [...] Read more.The distortions in the high-order structure of therapeutic monoclonal antibodies (mAbs) under different environmental conditions acutely affect mAb stability, resulting in altered safety, efficacy, and shelf-life profiles. The overall stability of mAbs depends on many factors, and it requires complementary techniques for an in-depth analysis. The stability of mAbs can be characterized by differential centrifugal sedimentation (DCS), differential scanning calorimetry (DSC), differential scanning fluorimetry (DSF), and size exclusion chromatography (SEC) techniques. In this report, temperature-ramped dynamic light scattering (DLS), and circular dichroism (CD) spectroscopy were employed as complementary tools to show how temperature and pH affect the aggregation of a model mAb, trastuzumab, in solution. The results showed that the aggregation onset temperature of trastuzumab defined by DLS was 75 °C, which decreases the amount of β-sheets and causes a slight increase in helix structures. Moreover, the melting temperature of trastuzumab was determined to be between 80–83 °C by temperature-ramped CD spectrophotometry, which is in line with the Tm of trastuzumab’s Fab region tested with DSC. Thus, unfolding and aggregation of trastuzumab start simultaneously at 75 °C, and unfolding triggers the aggregation. The temperature-ramped CD and DLS methods are robust tools to determine the thermal behavior of biosimilars in various solution conditions. Their complementary usage provides solid scientific background for regulatory applications and a better understanding of mAb instability and its relationship with structural changes.Full article►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 (a) The wavelength CD spectrum of 0.2 mg·mL−1 trastuzumab at 20 °C. (b) Temperature-wavelength CD spectrum of 0.2 mg·mL−1 trastuzumab between 65–90 °C by 2 °C intervals at 218 nm (c) Denaturation curve of trastuzumab at 202 nm. (d) Denaturation curve of trastuzumab at 218 nm. Full article ">Figure 2 (a) MRE values for trastuzumab within different pH at 20 °C. (b) Temperature-ramped CD spectrum of trastuzumab at pH 4 and (c) pH 9. Full article ">Figure 3 Temperature-dependent percentage change of antiparallel β sheets at different pH. Full article ">Figure 4 Estimated secondary structure subunits of trastuzumab at various temperatures. The data were collected with CD and processed with BeStSel [15] online tool. Full article ">Figure 5 (a) The corresponding hydrodynamic diameters to antibody concentrations between 0.25–16 mg·mL−1. Red dots represent the hydrodynamic sizes of trastuzumab at different concentrations. Inset: The intensity-based size measurement (d·nm) of trastuzumab at 20 °C. (b) Intensity-based hydrodynamic diameter change of 0.25, 0.5, 1, 2, 4, 8, 16 mg·mL−1 trastuzumab with 5 °C tem-perature ramps within 25–90 °C. (c) The temperature-ramped DLS measurement of 0.2 mg·mL−1 trastuzumab within the temperature range of 60–90 °C by varying 5 °C steps. (d) The intensity-based hydrodynamic diameter changes of trastuzumab at 70–75–80 °C. Full article ">Figure 6 (a) Volume-based and (b) number-based temperature-ramped hydrodynamic diameter change of trastuzumab at different pH values. Full article ">Scheme 1 Overview of thermal stability analysis of mAbs by complementary tools: CD and DLS (instability pathway image was adapted from Ref. [3], and IgG1 structure was adapted from Ref. [9]). Full article "> get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessReviewNanocarrier Systems in Taste MaskingbyNasr Eldin Hussein Nasr, Aliaa Nabil ElMeshad and Ahmed Roshdy Fares Sci. Pharm. 2022, 90(1), 20; https://doi.org/10.3390/scipharm90010020 - 04 Mar 2022Abstract Taste is the most crucial organoleptic parameter affecting patient compliance in the case of drugs with poor palatability. Taste masking is a major challenge for the development of orally ingested active pharmaceutical constituents in the pharmaceutical industry. Numerous conventional taste-masking techniques have been [...] Read more.Taste is the most crucial organoleptic parameter affecting patient compliance in the case of drugs with poor palatability. Taste masking is a major challenge for the development of orally ingested active pharmaceutical constituents in the pharmaceutical industry. Numerous conventional taste-masking techniques have been extensively studied. In parallel, affecting the drug solubility or release is a major concern of conventional taste-masking techniques. Recently, many nanocarrier systems have been introduced, claiming the advantage of effective taste masking without affecting either the drug solubility or its release. In this review, we will present new techniques for taste masking, including taste-masking techniques utilizing nanocarrier systems such as liposomes, polymeric and solid lipid nanoparticles, polymeric micelles, submicron lipid emulsions, and nanogels. We will chiefly highlight the composition of these systems and their applications in designing oral therapeutic delivery systems successful in masking the taste of bitter molecules.Full article►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 Study selection flow diagram. Full article ">Figure 2 Number of publications for nanocarrier systems in taste masking per year. Full article ">Figure 3 Different nanocarrier systems used for taste masking. Full article "> attachmentSupplementary material:Supplementary File 1 (ZIP, 387 KiB)get_appsubjectView online as:Abstract PageFull-Text HTMLOpen AccessArticleApplication of Quality by Design Approach to the Pharmaceutical Development of Anticancer Crude Extracts of Crocus sativus PerianthbyOlha Mykhailenko, Liudas Ivanauskas, Ivan Bezruk, Vilma Petrikaitė and Victoriya Georgiyants Sci. Pharm. 2022, 90(1), 19; https://doi.org/10.3390/scipharm90010019 - 03 Mar 2022Abstract The application of the Quality by Design (QbD) concept to extracts obtained from Crocus sativus perianth with potential anticancer activity will ensure the safety, efficiency, and quality control of the entire technological process, as well as determine the critical factors affecting the quality [...] Read more.The application of the Quality by Design (QbD) concept to extracts obtained from Crocus sativus perianth with potential anticancer activity will ensure the safety, efficiency, and quality control of the entire technological process, as well as determine the critical factors affecting the quality of extracts. Potentially critical points of the production of the plant extracts, including the cultivation and processing of the plant materials, the extraction process, and the choice of solvents, were identified using the Ishikawa diagram and FMEA risk assessment methods as well as the corrective actions proposed. The Herbal Chemical Marker Ranking System (HerbMars) approach was used to justify the Q-markers choice of Crocus, which takes into account bioavailability, pharmacological activity, and the presence of the selected standard. An experimental design (DoE) was used to assess the influence of potentially critical factors on the efficiency of the compound extraction from raw materials with water or ethanol. The presence of 16 compounds in Crocus perianth was determined by HPLC and their quantitative assessment was established. Selected compounds (ferulic acid, mangiferin, crocin, rutin, isoquercitrin) can be used for the quality control of Crocus perianth. In addition, the stigmas from the Volyn region met the requirements of ISO 3632 for saffron as a spice (category I). The cytotoxic activity against melanoma (IGR39) and triple-negative breast cancer (MDA-MB-231) cell lines of the hydroethanolic extract of C. sativus perianth was significantly more pronounced than the water extract, probably due to the chemical composition of the constituent components. The results show that the QbD approach is a powerful tool for process development for the production of quality herbal drugs.Full article►▼Show Figuresdiv" data-cycle-log="false">Figure 1Figure 1 The QbD approach to quality assurance in Crocus perianth drugs production. Adopted by [5,6,19]. Full article ">Figure 2 Risk assessment of C. sativus perianth extracts obtaining using the Ishikawa method. Full article ">Figure 3 DoE of obtaining C. sativus crude extracts with potential anticancer activity [14,17,25,30,31]. Full article ">Figure 4 Herb MaRS criteria for chosen of Q-markers for Crocus perianth raw material and crude extracts. Full article ">Figure 5 The chromatogram of C. sativus perianth fingerprints obtained by HPLC-DAD method at 270 nm (line 1), 310 nm (line 2), and 440 nm (line 3): mangiferin (A), isoorienthin (B), rutin (C), ferulic acid (D), isoquercitrin (E), tectoridin (F), crocin 4 (G), apigenin-7-glucoside (H), quercetin (I), nigricin (J), iristectorignin B (K), kaempferol (L). Full article ">Figure 6 Chemical structure of selected quality markers and some major compounds of C. sativus perianth. Full article ">Figure 7 EC50 values of C. sativus perianth extracts against MDA-MB-231 and IGR39 cell lines after 72 h, * p < 0.05, n = 3. Full article "> More Articles...Submit to Sci. Pharm. Review for Sci. Pharm.ShareJournal Menu►▼Journal MenuSci. Pharm. 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