Pharmaceuticals

13 pages, 1356 KiB

Open AccessFeature PaperArticle

Methanolic Extract of the Herb Ononis spinosa L. Is an Antifungal Agent with no Cytotoxicity to Primary Human Cells

by Dejan Stojković, Maria Inês Dias, Danijela Drakulić, Lillian Barros, Milena Stevanović, Isabel C. F. R. Ferreira and Marina D. Soković

Pharmaceuticals 2020, 13(4), 78; https://doi.org/10.3390/ph13040078 - 24 Apr 2020

Cited by 27 | Viewed by 4964

Abstract

Ononis spinosa L. is a plant traditionally used as folk remedy. There are numerous studies regarding chemical constituents and health beneficial properties of Ononidis Radix. The following study was designed to investigate chemical composition and antifungal potential of the methanolic extract obtained from [...] Read more.

Ononis spinosa L. is a plant traditionally used as folk remedy. There are numerous studies regarding chemical constituents and health beneficial properties of Ononidis Radix. The following study was designed to investigate chemical composition and antifungal potential of the methanolic extract obtained from the O. spinosa L. herb. Chemical analyses regarding phenolic compounds of O. spinosa were performed by liquid chromatography with mass spectrometry (LC-DAD-ESI/MSn). Antifungal activity, antibiofilm properties and antifungal mode of action of the extract were evaluated, as well as cytotoxicity. Chemical analyses revealed the presence of flavonoids, isoflavonoids and phenolic acids in O. spinosa, with kaempherol-O-hexoside-pentoside being the most abundant compound (5.1 mg/g extract). Methanolic extract was active against all of the tested microfungi with Penicillium aurantiogriseum being the most sensitive to the extract inhibitory effect at 0.02 mg/mL; and effectively inhibited biofilms formed by Candida strains. Minimum fungicidal concentrations of extract rose in the presence of ergosterol and leakage of cellular components was detected. The extract showed no cytotoxicity to human gingival fibroblast (HGF-1) cells. This study significantly contributes to overall knowledge about medicinal potential of O. spinosa herbal extract and enlightens previously unrevealed properties. O. spinosa aerial parts seem to be an interesting candidate for the development of antifungal preparations, non-toxic to human cells. Full article

(This article belongs to the Special Issue Bioactive Compounds from Plants and Foods with Pharmaceutical Interest)

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Retention time (Rt), wavelengths of maximum absorption in the visible region (λmax), mass spectral data and tentative identification of the phenolic compounds present in Ononis spinosa L., recorded at 280 nm (A) and 370 nm (B). Full article ">Figure 2
Minimum fungicidal concentrations (MFC) of O. spinosa methanolic extract against C. albicans in the presence of different ergosterol concentrations. There was no statistically significant difference between control sample and sample with added 25 µg/mL of ergosterol, while there was spastically significant difference between control a, 25 a µg/mL and 50 b µg/mL and 100 c µg/mL of added ergosterol (p < 0.05), a, b, c statistically significant difference between samples. Full article ">Figure 3
Leakage of cellular components recorded by absorbance at 260 and 280 nm in C. albicans treated with O. spinosa methanolic extract at 2 × MIC (minimum inhibitory concentration). There has been significant difference between control and treated samples (p < 0.05). Full article ">Figure 4
(A) Relative growth rate (%) of human gingival fibroblast (HGF-1) cells treated with different concentrations of O. spinosa extract showing no statistical difference between treatments (p < 0.05); (B) representative phase contrast image of non-treated control cells and (C) representative phase contrast image of cells treated with O. spinosa extract (400 µg/mL). Full article ">

17 pages, 2672 KiB

Open AccessArticle

Drug Conjugates for Targeting Eph Receptors in Glioblastoma

by Puja Sharma, Callie Roberts, Denise Herpai, Izabela D. Fokt, Waldemar Priebe and Waldemar Debinski

Pharmaceuticals 2020, 13(4), 77; https://doi.org/10.3390/ph13040077 - 23 Apr 2020

Cited by 10 | Viewed by 5497

Abstract

Glioblastoma (GBM) is a complex and heterogeneous tumor that warrants a comprehensive therapeutic approach for treatment. Tumor-associated antigens offer an opportunity to selectively target various components of the GBM microenvironment while sparing the normal cells within the central nervous system. In this study, [...] Read more.

Glioblastoma (GBM) is a complex and heterogeneous tumor that warrants a comprehensive therapeutic approach for treatment. Tumor-associated antigens offer an opportunity to selectively target various components of the GBM microenvironment while sparing the normal cells within the central nervous system. In this study, we conjugated a multivalent vector protein, QUAD 3.0, that can target four receptors: EphA3, EphA2, EphB2, and also IL-13RA2, spanning virtually 100% of the GBM microenvironment, to doxorubicin derivatives. The conjugates effectively bound to all four receptors, although to varying degrees, and delivered cytotoxic loads to both established and patient-derived GBM cell lines, with IC₅₀ values in the low nM range. The conjugates were also non-toxic to animals. We anticipate that the QUAD 3.0 Dox conjugates will be further used in preclinical models and possibly clinics in the foreseeable future. Full article

(This article belongs to the Special Issue Targeting the Eph–ephrin System)

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Dox derivatives conjugation to QUAD 3.0. (a) A schematic of the QUAD 3.0 protein showing the CH2 CH3 domains of the human IgG1 used as a scaffold. IL-13.E13K, a modified version of the IL-13 ligand, is present in the N-terminal of the molecule and ephrinA5 along with a cysteine present in the C-terminal end of the molecule; (b) A schematic of the conjugation of Dox derivatives to the QUAD 3.0 molecule; (c) The structure of the three Dox derivatives used in the study that are thiol reactive (circled) and form a stable thioether bond with the thiol present in the cysteine residue; (d) SDS-PAGE (left panel) and corresponding fluorescent image the same gel (right panel) of QUAD 3.0 (lane 1) and its QUAD 3.0-WP936 conjugate before (lane 2) and after the removal of excess unconjugated WP936 using a Zeba desalting column (lane 3). To detect the fluorescently labeled protein, the gel was scanned using an Amersham AI 600RGB digital scanner. Full article ">Figure 2
QUAD 3.0-WP936 binding to EphA3, EphA2, EphB2, and IL-13RA2 receptors. ELISA assay showing the binding of unconjugated QUAD 3.0 and QUAD 3.0-WP936 conjugate to (a) EphA3; (b) EphA2; (c) EphB2; and (d) IL-13RA2 receptor proteins. Full article ">Figure 3
QUAD 3.0-WP1737 binding to EphA3, EphA2, EphB2, and IL-13RA2 receptors. ELISA assay showing the binding of unconjugated QUAD 3.0 and QUAD 3.0-WP1737 conjugate to (a) EphA3; (b) EphA2; (c) EphB2; and (d) IL-13RA2 receptor proteins. Full article ">Figure 4
QUAD 3.0 Dox 1244 binding to EphA3, EphA2, EphB2, and IL-13RA2 receptors. ELISA results showing the binding of unconjugated QUAD 3.0 and QUAD 3.0-WP1244 conjugate to (a) EphA3; (b) EphA2; (c) EphB2; and (d) IL-13RA2 receptor proteins. Full article ">Figure 5
QUAD 3.0-WP936 binding and internalization to U-251 GBM cells. The cells were treated with 1.0–5.0 μg of the conjugate for 4 h. WP936 was detected by red fluorescence in a TRITC channel and QUAD 3.0 by green fluorescence through Alexa-488-conjugated anti-human IgG. Sections were counterstained for DAPI (blue fluorescence) (scale bar: 50 μm). Full article ">Figure 6
QUAD 3.0-Dox conjugates are cytotoxic to established and patient-derived GBM cell lines. MTT assay of (a) unconjugated WP936 and QUAD 3.0-WP936 conjugates in (i) U-251, (ii) BTCOE 4795, and (iii) T98G cells; (b) unconjugated WP1737 and QUAD 3.0-WP1737 conjugates in (i) U-251, (ii) BTCOE 4795, and (iii) T98G cells; and (c) unconjugated WP1244 and QUAD 3.0-WP1244 conjugates in (i) U-251, (ii) BTCOE 4795, and (iii) T98G cells. Full article ">Figure 7
QUAD 3.0-WPD936 is safe in mice. Change in weight (%) of C57BL/6 mice after intracranial injections of QUAD 3.0-WP936. Full article ">

21 pages, 2749 KiB

Open AccessReview

Atomic Nanogenerators in Targeted Alpha Therapies: Curie’s Legacy in Modern Cancer Management

by Mareike Roscher, Gábor Bakos and Martina Benešová

Pharmaceuticals 2020, 13(4), 76; https://doi.org/10.3390/ph13040076 - 23 Apr 2020

Cited by 31 | Viewed by 5863

Abstract

Atomic in vivo nanogenerators such as actinium-225, thorium-227, and radium-223 are of increasing interest and importance in the treatment of patients with metastatic cancer diseases. This is due to their peculiar physical, chemical, and biological characteristics, leading to astonishing responses in otherwise resistant [...] Read more.

Atomic in vivo nanogenerators such as actinium-225, thorium-227, and radium-223 are of increasing interest and importance in the treatment of patients with metastatic cancer diseases. This is due to their peculiar physical, chemical, and biological characteristics, leading to astonishing responses in otherwise resistant patients. Nevertheless, there are still a few obstacles and hurdles to be overcome that hamper the broader utilization in the clinical setting. Next to the limited supply and relatively high costs, the in vivo complex stability and the fate of the recoiling daughter radionuclides are substantial problems that need to be solved. In radiobiology, the mechanisms underlying treatment efficiency, possible resistance mechanisms, and late side effect occurrence are still far from being understood and need to be unraveled. In this review, the current knowledge on the scientific and clinical background of targeted alpha therapies is summarized. Furthermore, open issues and novel approaches with a focus on the future perspective are discussed. Once these are unraveled, targeted alpha therapies with atomic in vivo nanogenerators can be tailored to suit the needs of each patient when applying careful risk stratification and combination therapies. They have the potential to become one of the major treatment pillars in modern cancer management. Full article

(This article belongs to the Special Issue The Future Direction of Radiopharmaceutical Development for Cancer Theranostics)

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Graphical abstract

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Schematic representation of the atomic in vivo nanogenerator 225Ac (τ½ = 9.9 d, Eα = 5.8 MeV). 225Ac decays through four net α-disintegrations (five in total) and two net β–-disintegrations (three in total) into stable 209Bi. The 225Ac decay chain possess two eligible γ-emissions for detection, 218 keV (I = 11.4%, 221Fr) and 440 keV (I = 25.9%, 213Bi). The most prominent daughter radionuclide is 213Bi (τ½ = 45.6 min, Eα = 5.9 MeV), which is also used for targeted alpha therapy (TAT) itself. The half-life (τ½), known energies connected to recoil events (translational kinetic energy Et), and the decay energies (Eα, Eβ, Eγ) are indicated on the scheme. Data were derived from Nucleonica GmbH, Nuclide Datasheets, Nucleonica Nuclear Science Portal (<a href="http://www.nucleonica.com" target="_blank">www.nucleonica.com</a>), Version 3.0.65, Karlsruhe (2017). Full article ">Figure 2
Schematic representation of the atomic in vivo nanogenerator 227Th (τ½ = 18.7 d, Eα = 6.0 MeV). 227Th decays through five net α-disintegrations (six in total) and two net β–-disintegrations (three in total) into stable 207Pb. 227Th possess at least four eligible γ-emissions for detection, 235 keV (I = 12.9%, 227Th), 269 keV (I = 13.9%, 223Ra), 405 keV (I = 3.8%, 211Pb), and 351 keV (I = 13.0%, 211Bi). The most prominent daughter radionuclide is 223Ra (τ½ = 11.4 d, Eα = 6.3 MeV), which is also used for TAT itself. The half-life (τ½), known energies connected to recoil events (translational kinetic energy Et), and the decay energies (Eα, Eβ, Eγ) are indicated on the scheme. Data were derived from Nucleonica GmbH, Nuclide Datasheets, Nucleonica Nuclear Science Portal (<a href="http://www.nucleonica.com" target="_blank">www.nucleonica.com</a>), Version 3.0.65, Karlsruhe (2017). Full article ">Figure 3
Structure of the current state-of-the-art chelating agent DOTA, next to the structures of the promising chelating agents Macropa for 225Ac coordination and Me-3,2-HOPO for 227Th coordination. Each of these chelators offers broad derivatization possibilities to further alter conjugation, labeling efficiency, and complex stability. Coordinating nitrogen donors are highlighted in blue, and oxygen donors in red. DOTA: 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid. Full article ">Figure 4
Nuclear recoil effect during α-decay within atomic in vivo nanogenerators. Schematic representation of the conservation of momentum law describing the transfer of the decay energy to the α-particle and daughter nucleus. The recoiling daughter nucleus can move up to ≈100 nm with each single α-decay and, hence, could travel considerable distances in a water-like environment (e.g., tissue). Note: The position of the α-particle is to show direction and not position. Full article ">Figure 5
Nuclear recoil effect during α-decay within pharmaceuticals for targeted alpha therapy (TAT-P). Schematic representation of two hypothetical scenarios describing the fate of the recoiling daughter radionuclide that gets released from the chelating moiety of TAT-P in vivo. The upper section labelled “No TAT-P internalization” depicts a daughter radionuclide that is released into the blood stream while causing either unspecific local damage to healthy tissue or travels further with the blood stream and causes analogical damage distantly elsewhere. The lower section labelled “TAT-P internalization” depicts TAT-P that specifically internalizes into the targeted tumor cell. The daughter radionuclide is then released with a high probability inside the tumor cell or to a minor extent might escape the tumor cell and cause damage not only to the target tumor cell but, depending on the travelled distance, to other cells as well. Full article ">Figure 6
Possible outcomes of targeted alpha therapies. Prostate-specific membrane antigen (PSMA)-targeted alpha therapy of metastasized castration-resistant prostate cancer (mCRPC) was selected as an example for the schematic representation of three main hypothetical scenarios. These are (a) full and long-lasting remission, (b) disease recurrence after the successful outcome of the initial therapy, and (c) disease progression, which occurs in spite of the applied therapy (therapy resistance). In addition, possible early and late side effects, such as salivary gland, kidney, and bladder damage, are also indicated in the figure. TAT is usually applied as a third-line or salvage therapy when all other previous treatment options have failed. Full article ">

10 pages, 1927 KiB

Open AccessReview

Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Ubiquitylation as a Novel Pharmaceutical Target for Cystic Fibrosis

by Ryosuke Fukuda and Tsukasa Okiyoneda

Pharmaceuticals 2020, 13(4), 75; https://doi.org/10.3390/ph13040075 - 22 Apr 2020

Cited by 15 | Viewed by 7715

Abstract

Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene decrease the structural stability and function of the CFTR protein, resulting in cystic fibrosis. Recently, the effect of CFTR-targeting combination therapy has dramatically increased, and it is expected that add-on drugs that [...] Read more.

Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene decrease the structural stability and function of the CFTR protein, resulting in cystic fibrosis. Recently, the effect of CFTR-targeting combination therapy has dramatically increased, and it is expected that add-on drugs that modulate the CFTR surrounding environment will further enhance their effectiveness. Various interacting proteins have been implicated in the structural stability of CFTR and, among them, molecules involved in CFTR ubiquitylation are promising therapeutic targets as regulators of CFTR degradation. This review focuses on the ubiquitylation mechanism that contributes to the stability of mutant CFTR at the endoplasmic reticulum (ER) and post-ER compartments and discusses the possibility as a pharmacological target for cystic fibrosis (CF). Full article

(This article belongs to the Special Issue Targeted Protein Degradation: From Chemical Biology to Drug Discovery)

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Wild type (WT)–cystic fibrosis transmembrane conductance regulator (CFTR) is translated at the ribosome on the endoplasmic reticulum (ER), and the membrane spanning domain (MSD) penetrates the ER membrane. Upon translation, each domain of the CFTR is folded with the assistance of chaperone–cochaperone complexes on the cytoplasmic side and inside of the ER to form a metastable structure (Co-translational folding). After the translation is completed, the domains interact by the action of the chaperones to form a native structure (Post-translational folding). Found in the nucleotide binding domain 1 (NBD1) of WT–CFTR, the ER export motif is exposed and CFTR is transported from the ER to the Golgi apparatus by coat protein complex II (COP II) vesicles. Full article ">Figure 2
The stability of the interaction between nucleotide binding domain 1(NBD1) and membrane spanning domains (MSD1,2) decreases due to the NBD1 structural abnormality in the ∆F508–cystic fibrosis transmembrane conductance regulator (CFTR) mutation. The CFTR-associated E3 ligases recognize the misfolded regions mediated by chaperones and facilitate the co-translational and post-translational ubiquitylation. The endoplasmic reticulum (ER) export motif is folded inside ∆F508–CFTR and, conversely, it stays in the ER due to the exposure of the ER retention motif. Ubiquitylated ∆F508–CFTR is ultimately degraded by proteasome via retrotranslocation to the cytoplasm by the Derlin1– valosin-containing protein(VCP) complex. Full article ">Figure 3
Wild-type (WT)– cystic fibrosis transmembrane conductance regulator (CFTR) exerts a stable channel function on the plasma membrane (PM). Internalized WT–CFTR could undergo deubiquitylation by ubiquitin Specific Protease 10 (USP10) and, consequently, be recycled to the PM. When P. aeruginosa infection occurs, USP10 is inhibited by CFTR inhibitory factor (Cif) produced by the bacteria, and CFTR turnover is accelerated. The misfolded region of ∆F508–CFTR is recognized by a chaperone complex at the PM. The chaperone complex helps to maintain the channel function of ∆F508–CFTR at the PM but promotes the CFTR elimination from the PM by facilitating the C-terminal Hsp-interacting protein (CHIP)-mediated ubiquitylation. Rififylin (RFFL) is localized at the PM and endosome and promotes endocytosis of ∆F508–CFTR by directly recognizing misfolded-nucleotide binding domain 1 (NBD1) and facilitating K63-linked poly-ubiquitylation. The endocytosed ΔF508–CFTR is invaded into the intraluminal vesicle (ILV) of the multivesicular body (MVB) by endosomal sorting complexes required for transport (ESCRT) proteins such as hepatocyte growth factor-regulated tyrosine kinase substrate (HGS, also known as Hrs) and tumor susceptibility gene 101 (TSG101) and sent to lysosomal degradation. Full article ">

19 pages, 12595 KiB

Open AccessReview

Selective Degradation of Target Proteins by Chimeric Small-Molecular Drugs, PROTACs and SNIPERs

by Minoru Ishikawa, Shusuke Tomoshige, Yosuke Demizu and Mikihiko Naito

Pharmaceuticals 2020, 13(4), 74; https://doi.org/10.3390/ph13040074 - 21 Apr 2020

Cited by 20 | Viewed by 5459

Abstract

New therapeutic modalities are needed to address the problem of pathological but undruggable proteins. One possible approach is the induction of protein degradation by chimeric drugs composed of a ubiquitin ligase (E3) ligand coupled to a ligand for the target protein. This article [...] Read more.

New therapeutic modalities are needed to address the problem of pathological but undruggable proteins. One possible approach is the induction of protein degradation by chimeric drugs composed of a ubiquitin ligase (E3) ligand coupled to a ligand for the target protein. This article reviews chimeric drugs that decrease the level of specific proteins such as proteolysis targeting chimeric molecules (PROTACs) and specific and nongenetic inhibitor of apoptosis protein (IAP)-dependent protein erasers (SNIPERs), which target proteins for proteasome-mediated degradation. We cover strategies for increasing the degradation activity induced by small molecules, and their scope for application to undruggable proteins. Full article

(This article belongs to the Special Issue Targeted Protein Degradation: From Chemical Biology to Drug Discovery)

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9 pages, 2233 KiB

Open AccessCommunication

Identification and Quantification of Stilbenes (Piceatannol and Resveratrol) in Passiflora edulis By-Products

by Karolline Krambeck, Ana Oliveira, Delfim Santos, Maria Manuela Pintado, João Baptista Silva, José Manuel Sousa Lobo and Maria Helena Amaral

Pharmaceuticals 2020, 13(4), 73; https://doi.org/10.3390/ph13040073 - 20 Apr 2020

Cited by 27 | Viewed by 5336

Abstract

Recently, studies on the by-products from the food industry, such as passion fruit seeds, have significantly increased, as these can have an added value, due to their properties, such as potential antioxidant activity. This study was conducted to determine the presence of piceatannol [...] Read more.

Recently, studies on the by-products from the food industry, such as passion fruit seeds, have significantly increased, as these can have an added value, due to their properties, such as potential antioxidant activity. This study was conducted to determine the presence of piceatannol and resveratrol in various extracts of passion fruit (Passiflora edulis) seeds from Madeira Island and a commercial passion fruit oil was used as reference. The commercial oil and the extracts that were obtained by traditional Soxhlet method with ethanol and acetone did not reveal the presence of the two stilbenes, piceatannol and resveratrol. However, the extracts that were obtained by the ultrasound method showed significant amounts of piceatannol and resveratrol when compared with the commercial oil. The presence of these compounds indicates that this oil could have potential application in cosmetic and pharmaceutical industries, due to their proven antioxidant and anti-aging properties. Full article

(This article belongs to the Special Issue Selected Papers from the 5th International Electronic Conference on Medicinal Chemistry)

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High-performance liquid chromatography with diode array detection (HPLC-DAD) chromatograms of the standard solutions: (a) piceatannol; (b) resveratrol. UV spectra of the standard solutions: (c) piceatannol; and, (d) resveratrol. Full article ">Figure 2
HPLC-DAD chromatograms (320 nm) of the extract obtained by the ultrasound method with ethanol (a) and with acetone (b). Number 1 and 2 corresponds to piceatannol and resveratrol, respectively. The UV spectra of piceatannol (red) and resveratrol (blue) detected on ethanol (c) and acetone (d) extracts. Full article ">Figure 3
Piceatannol and resveratrol content (μg/mL) in ethanol and acetone extracts obtained with ultrasound method. Results are expressed as Mean ± SD. Statistical comparisons were made using one-way ANOVA, followed by the Tukey’s multiple comparisons test. Values significantly different from piceatannol (**** p < 0.05). Full article ">Figure 4
Chromatograms of the commercial passion fruit oil. Full article ">Figure 5
Flow diagram for the extraction and separation by HPLC of piceatannol and resveratrol. Full article ">

2 pages, 315 KiB

Open AccessCorrection

Correction: McLoughlin, E.C.; O’Boyle, N.M. Colchicine-Binding Site Inhibitors from Chemistry to Clinic: A Review. Pharmaceuticals 2020, 13, 8

by Eavan C. McLoughlin and Niamh M. O’Boyle

Pharmaceuticals 2020, 13(4), 72; https://doi.org/10.3390/ph13040072 - 20 Apr 2020

Cited by 10 | Viewed by 2326

Abstract

We, the authors, wish to make the following corrections to our paper [...] Full article

(This article belongs to the Special Issue Recent Contributions in Medicinal Chemistry Within European GP2A Network)

14 pages, 350 KiB

Open AccessArticle

Antioxidant, Anti-Inflammatory, and Antidiabetic Activities of Leaves and Stems of Uapaca bojeri Bail. (EUPHORBIACEAE), an Endemic Plant of Madagascar

by Zoarilala Rinah Razafindrakoto, Dario Donno, Nantenaina Tombozara, Harilala Andriamaniraka, Charles Andrianjara, David Ramanitrahasimbola and Gabriele Loris Beccaro

Pharmaceuticals 2020, 13(4), 71; https://doi.org/10.3390/ph13040071 - 17 Apr 2020

Cited by 17 | Viewed by 4216

Abstract

Uapaca bojeri is an endemic Malagasy plant used by the local population. This work aimed to evaluate antioxidant, anti-inflammatory, and antidiabetic activities of the methanol extracts of U. bojeri leaves and stems and to report their total phenolic content and the bioactive compound [...] Read more.

Uapaca bojeri is an endemic Malagasy plant used by the local population. This work aimed to evaluate antioxidant, anti-inflammatory, and antidiabetic activities of the methanol extracts of U. bojeri leaves and stems and to report their total phenolic content and the bioactive compound content by HPLC methods. Antioxidant capacity was determined by DPPH and ferric reducing antioxidant power (FRAP) assays. An in vivo carrageenan-induced paw oedema and acetic acid-induced writhing test in mice were used for anti-inflammatory activity evaluation. An oral glucose tolerance test was performed in mice to evaluate antidiabetic activity. The total bioactive compound content of leaves was higher than that of stems. Stem methanol extract inhibited the free radical DPPH more than the leaf methanol extract. Leaf methanol extract inhibited, in a dose-dependent manner, the carrageenan-induced paw oedema more than the stem extract, but their inhibition of the pain symptoms caused an acetic acid-induced decrease similar to the number of writhes in the dose-dependent case. The leaf and stem methanol extracts significantly reduced blood glucose levels after 30 min of glucose loading in mice compared to the control group blood glucose reduction. The presence of several bioactive compounds in U. bojeri contributed to the different biological activities, but isolation and identification of these bioactive molecules are necessary to confirm these pharmacological properties. Full article

(This article belongs to the Special Issue Medicinal Plants 2020)

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11 pages, 1401 KiB

Open AccessArticle

Electroanalysis Applied to Compatibility and Stability Assays of Drugs: Carvedilol Study Case

by Murilo Ferreira de Carvalho, Luane Ferreira Garcia, Isaac Yves Lopes de Macedo, Ricardo Neves Marreto, Mayk Teles de Oliveira, Renê Oliveira do Couto, Carlos Eduardo Peixoto da Cunha, Karla Carneiro de Siqueira Leite, Kênnia Rocha Rezende, Fabio Bahls Machado, Vernon Somerset and Eric de Souza Gil

Pharmaceuticals 2020, 13(4), 70; https://doi.org/10.3390/ph13040070 - 17 Apr 2020

Cited by 3 | Viewed by 2609

Abstract

Carvedilol (CRV) is a non-selective blocker of α and β adrenergic receptors, which has been extensively used for the treatment of hypertension and congestive heart failure. Owing to its poor biopharmaceutical properties, CRV has been incorporated into different types of drug delivery systems [...] Read more.

Carvedilol (CRV) is a non-selective blocker of α and β adrenergic receptors, which has been extensively used for the treatment of hypertension and congestive heart failure. Owing to its poor biopharmaceutical properties, CRV has been incorporated into different types of drug delivery systems and this necessitates the importance of investigating their compatibility and stability. In this sense, we have investigated the applicability of several electroanalytical tools to assess CRV compatibility with lipid excipients. Voltammetric and electrochemical impedance spectroscopy techniques were used to evaluate the redox behavior of CRV and lipid excipients. Results showed that Plurol^® isostearic, liquid excipient, and stearic acid presented the greatest anode peak potential variation, and these were considered suitable excipients for CRV formulation. CRV showed the highest stability at room temperature and at 50 °C when mixed with stearic acid (7% w/w). The results also provided evidence that electrochemical methods might be feasible to complement standard stability/compatibility studies related to redox reactions. Full article

(This article belongs to the Section Pharmaceutical Technology)

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Chemical structure of carvedilol. Full article ">Figure 2
Cyclic voltammograms of CP control (─), CPPI20% (---), and CPPI30% (─) (A); CPSfO30% (─), CPSeO30% (•••), and CPCO30% (-•-•-) e CP control (─) (B). All experiments were performed in triplicate in 0.1 mol L−1 KCl, PBS, pH 7.0. Plurol® isostearic carbon paste (CPPI), safflower oil carbon paste (CPSfO), sesame oil carbon paste (CPSeO), canola oil carbon paste (CPCO). Full article ">Figure 3
Solid-state differential pulse voltammograms of 1% CRV in: CP control (grey line of <a href="#pharmaceuticals-13-00070-f003" class="html-fig">Figure 3</a>A–C) and in CPSeO30% (—) and CPCO30% (- - -) (A), CPCpt7% (- • - •) and CPEA7% (• • •) (B), CPPI20% without CRV (• • •) and with 1% CRV (- • • -) (C). All assays were performed in 0.1 mol L−1 KCl, PBS, pH 7.0. Sesame oil carbon paste (CPSeO), canola oil carbon paste (CPCO); Compritol® carbon paste (CPCpt); estearic acid carbon paste (CPSfO); Plurol® isostearic carbon paste (CPPl) and Carvedilol (CRV). Full article ">Figure 4
Electrochemical impedance Nyquist plots of CP control (▲). (A) CPPI20% (■) and (B) CPEA7% (●). Insert: the related cyclic voltammetry (CV) scans. CP control (──), CPPI20% (- - -), and CPEA7% (- • -). All CV and electrochemical impedance spectroscopy (EIS) assays were performed with 0.05 mol.L−1 potassium ferri/ferrocyanide in 0.1 mol L−1 KCl solution. Full article ">Figure 5
Results shown for CRV decay in different CPs stored at room temperature for up to 180 days. Full article ">Figure 6
Results shown for CRV decay in different CPs binary systems at a 50 °C (A) and under darkness for up to 10 days of storage (B). Full article ">

11 pages, 1029 KiB

Open AccessArticle

Evaluation of the Anti-Tumor Activity of Small Molecules Targeting Eph/Ephrins in APC ^min/J Mice

by Miriam Corrado, Carmine Giorgio, Elisabetta Barocelli, Giuseppe Vittucci Marzetti, Anna Maria Cantoni, Rosanna Di Lecce, Matteo Incerti, Riccardo Castelli, Alessio Lodola and Massimiliano Tognolini

Pharmaceuticals 2020, 13(4), 69; https://doi.org/10.3390/ph13040069 - 16 Apr 2020

Cited by 1 | Viewed by 2986

Abstract

The Eph receptors are the largest receptors tyrosine kinases (RTKs) family in humans and together with ephrin ligands constitute a complex cellular communication system often dysregulated in many tumors. The role of the Eph-ephrin system in colorectal cancer (CRC) has been investigated and [...] Read more.

The Eph receptors are the largest receptors tyrosine kinases (RTKs) family in humans and together with ephrin ligands constitute a complex cellular communication system often dysregulated in many tumors. The role of the Eph-ephrin system in colorectal cancer (CRC) has been investigated and different expression of Eph receptors have been associated with tumor development and progression. In light of this evidence, we investigated if a pharmacological approach aimed at inhibiting Eph/ephrin interaction through small molecules could prevent tumor growth in APC ^min/J mice. The 8-week treatment with the Eph-ephrin antagonist UniPR129 significantly reduced the number of adenomas in the ileum and decreased the diameter of adenomas in the same region. Overall our data suggested as UniPR129 could be able to slow down the tumor development in APC ^min/J mice. These results further confirm literature data about Eph kinases as a new valuable target in the intestinal cancer and for the first time showed the feasibility of the Eph-ephrin inhibition as a useful pharmacological approach against the intestinal tumorigenesis. In conclusion this work paves the way for further studies with Eph-ephrin inhibitors in order to confirm the Eph antagonism as innovative pharmacological approach with preventive benefit in the intestinal tumor development. Full article

(This article belongs to the Special Issue Targeting the Eph–ephrin System)

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Trend of mice weight over the study. The mean weight of mice, recorded once a week in each experimental group (n = 7), was represented. Full article ">Figure 2
(a) Comparison of the total number of adenomas developed in the intestine between control and treatments groups; (b) Comparison of the number of adenomas developed in the ileum section between control and treatments groups. n = 7. Full article ">Figure 3
Comparison of the number of tumors developed in different sections of the intestine between control and UniPR129 group. Two-way ANOVA followed by Bonferroni’s post-test was used to compare control to UniPR129 group. *** p < 0.001, n = 7. Full article ">Figure 4
Diameter of all the adenomas collected in the Ileum of APC min/J CTR mice and after oral administration of UniPR129 30 mg/kg. Mann–Whitney test was performed to compare tumor diameter of control to UniPR129 group (*** p < 0.001). Black dots: control; red squares: UniPR129. Full article ">Figure 5
Adenomas dimensional distribution for control and UniPR129 group. Two-way ANOVA followed by Bonferroni’s post-test was used to compare control group to UniPR129 group. * p < 0.05, n = 7. Full article ">

12 pages, 975 KiB

Open AccessCommunication

In-Vitro Evaluation of Antioxidant, Antiproliferative and Photo-Protective Activities of Benzimidazolehydrazone Derivatives

by Anna Baldisserotto, Monica Demurtas, Ilaria Lampronti, Massimo Tacchini, Davide Moi, Gianfranco Balboni, Silvia Vertuani, Stefano Manfredini and Valentina Onnis

Pharmaceuticals 2020, 13(4), 68; https://doi.org/10.3390/ph13040068 - 15 Apr 2020

Cited by 16 | Viewed by 3152

Abstract

In the search of multifunctional compounds we designed benzimidazole derivatives endowed with phenolic hydroxy groups and a hydrazone moiety as potential radical-scavenger and the antioxidant agents. The target molecules have been prepared by a simple synthetic procedure and tested for their antioxidant activity [...] Read more.

In the search of multifunctional compounds we designed benzimidazole derivatives endowed with phenolic hydroxy groups and a hydrazone moiety as potential radical-scavenger and the antioxidant agents. The target molecules have been prepared by a simple synthetic procedure and tested for their antioxidant activity by DPPH, FRAP, and ORAC test, for photoprotective activity against UV rays and for antiproliferative activity against Colo-38 melanoma cells. Furthermore, two different dermocosmetic formulations were prepared with the compounds endowed with the best antioxidant and photoprotective profile and their release from formulation evaluated using Franz Cells system. High antioxidant activity is related to the presence of at least two hydroxy groups on arylidene moiety of benzimidazoles. Structure activity analysis revealed that the position of hydroxy groups is crucial for antioxidant activity as well as the presence of a 2-hydroxy-4-(diethylamino)arylidene group. The same correlation pattern was found to be related to photoprotective activity resulting in an UVA Protection Factor better than the commercial solar filter PBSA and antiproliferative activity against melanoma cells without producing cytotoxicity on normal keratinocytes. The release analysis indicated that high antioxidant activities are achieved with limited release at concentration compatible with the use as UV sunscreen filter. Full article

(This article belongs to the Section Medicinal Chemistry)

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11 pages, 1050 KiB

Open AccessArticle

Oral l-Cysteine Supplementation Enhances the Long Term-Effect of Topical Basic Fibroblast Growth Factor (bFGF) in Reducing the Corneal Haze after Photorefractive Keratectomy in Myopic Patients

by Alessandro Meduri, Loredana Bergandi, Pietro Perroni, Francesca Silvagno and Pasquale Aragona

Pharmaceuticals 2020, 13(4), 67; https://doi.org/10.3390/ph13040067 - 15 Apr 2020

Cited by 7 | Viewed by 3127

Abstract

We aimed at evaluating the long-term effects of l-cysteine oral supplementation to basic fibroblast growth factor (bFGF) eye-drops on corneal re-epithelization and transparency in myopic patients subjected to photorefractive keratectomy (PRK). Forty patients subjected to bilateral PRK for myopia were enrolled and [...] Read more.

We aimed at evaluating the long-term effects of l-cysteine oral supplementation to basic fibroblast growth factor (bFGF) eye-drops on corneal re-epithelization and transparency in myopic patients subjected to photorefractive keratectomy (PRK). Forty patients subjected to bilateral PRK for myopia were enrolled and randomly divided into two groups receiving an additional therapy together with the standard postoperative treatment consisting in local tobramycin 0.3%, dexamethasone 0.1%, diclofenac 0.1%, and 0.2% hyaluronate. Group 1 included 20 patients (11 males and 9 females; 34.09 ± 8 years of age) receiving only bFGF eye-drops (10 μg/10 μL) four times a day for 7 days starting from the day of surgery; Group 2 included 20 patients (12 males and 8 females; 37.35 ± 11.5 years of age) who were postoperatively administered with topical basic fibroblast growth factor (bFGF; 10 μg/10 μL) four times a day for 7 days plus oral l-cysteine supplementation (500 mg/capsule) once a day for 15 days, starting 7 days before PRK. Patients were followed-up for 12 months. Clinical ophthalmologic parameters were recorded for all the 80 examined eyes. The corneal transparency was evaluated in vivo by slit lamp and confocal microscopy. The data showed that: (a) the corneal haze occurred in a smaller percentage of the patients who were postoperatively administered with topical bFGF plus oral l-cysteine supplementation (Group 2) compared to patients who received only bFGF (Group 1); (b) at 6 months of follow-up, the stromal mean image brightness of the patients belonging to Group 2 was significantly lower than that of the Group 1 (p < 0.03), and, interestingly, the difference was even more evident at 12 month from the treatment (p < 0.001). Moreover, the final mean of the spherical equivalent refraction was −0.06 ± 0.2 D in Group 1 and −0.08 ± 0.3 D in Group 2, whereas the final uncorrected distance visual acuity (UDVA) was equal or superior to 20/25 in 100% of eyes in both Group 1 and 2. Post refractive patients can benefit from the administration of l-cysteine before the surgery and in association with bFGF in the early postoperative period, showing a faster corneal re-epithelization able to prevent corneal haze in the long-term recovery. Full article

(This article belongs to the Special Issue Advances in Ocular Pharmacology)

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Figure 1
Corneal haze incidence at 3, 6, and 12 month of follow-up as a function of its time of regression in patients who received only bFGF (bFGF Group) and in patients who were administered topical bFGF plus oral l-cysteine supplementation (bFGF + l-cys Group). * p < 0.05 bFGF + l-cys Group vs. bFGF Group at each time of follow-up. Full article ">Figure 2
Mean image brightness of the stroma (marker of altered cornea), measured with the ConfoScan 4 confocal microscope and expressed as mean ± SD scatter units (SU) in corneas of patients who received only bFGF (bFGF Group) and of patients who were administered topical bFGF plus oral l-cysteine supplementation (bFGF + l-cys Group). ○ p < 0.05 bFGF Group at 3, 6, and 12 months of follow-up vs. bFGF at 1 month of follow-up; ●● p < 0.01 and ●●● p < 0.001 bFGF + l-cys Group at 3, 6, and 12 month of follow-up vs. bFGF + l-cys Group at 1 month of follow-up; * p < 0.03 and ** p < 0.001 bFGF + l-cys Group vs. bFGF Group at each time of follow-up. Full article ">Figure 3
Representative ConfoScan 4 confocal microscope images of the stroma of 5 patients who received only bFGF (bFGF Group) and of 5 patients who were administered topical with bFGF plus oral l-cysteine supplementation (bFGF + l-cys Group). Images were taken at 6 months of follow-up. Yellow arrows indicate the light scattering regions of cornea. Full article ">

10 pages, 1635 KiB

Open AccessFeature PaperArticle

Synthesis of Silver Nanoparticles Using Odontosoria chinensis (L.) J. Sm. and Evaluation of their Biological Potentials

by Marimuthu alias Antonysamy Johnson, Thangaiah Shibila, Santhanam Amutha, Irwin R. A. Menezes, José G. M. da Costa, Nadghia F. L. Sampaio and Henrique D. M. Coutinho

Pharmaceuticals 2020, 13(4), 66; https://doi.org/10.3390/ph13040066 - 13 Apr 2020

Cited by 15 | Viewed by 3381

Abstract

The present study was aimed to synthesize silver nanoparticles (AgNPs) from the aqueous extracts of Odontosoria chinensis (L.) J. Sm. and the synthesized AgNPs were examined for their biopotentials. The Odontosoria chinensis extracts were added to 1 mM AgNO₃ solution with different [...] Read more.

The present study was aimed to synthesize silver nanoparticles (AgNPs) from the aqueous extracts of Odontosoria chinensis (L.) J. Sm. and the synthesized AgNPs were examined for their biopotentials. The Odontosoria chinensis extracts were added to 1 mM AgNO₃ solution with different ratios viz., 0.5:9.5, 1:9, 1.5:8.5 and 2:8 ratios for the reduction of Ag ions. After reduction, the AgNPs of Odontosoria chinensis were analyzed spectroscopically for further confirmation. The synthesized AgNPs of Odontosoria chinensis were characterized by pH, ultra violet–visible spectroscopy (UV-Vis), Fourier transform–infra red spectroscopy (FT-IR), scanning electron microscopy-energy dispersive X-ray analysis (SEM-EDAX) and X-Ray diffraction (XRD). The time taken for the complete reduction of Silver (Ag) in solution to nanoparticle was 10 min. The O. chinensis aqueous extracts mediated silver nanoparticles showed a broad peak with distinct absorption at around 400–420 nm and confirmed the silver nanoparticle formation. FT-IR results also confirmed the existence of organic materials in the silver nanoparticles of O. chinensis. The EDX spectra of AgNPs of O. chinenesis revealed the occurrence of a strong Ag peak. The synthesis of AgNPs of O. chinenesis was confirmed with the existence of a peak at 46.228°. The toxic potential of AgNPs of O. chinenesis showed varied percentage mortality with the LC₅₀ values of 134.68 μL/ 50 mL and 76.5 μL/50 mL, respectively. The anti-inflammatory and anti-diabetic activities of aqueous and AgNPs of O. chinenesis were statistically significant at p < 0.05 level. Conclusion: The results demonstrated the toxicity, anti-diabetic and anti-inflammatory potential of the studied AgNPs. The synthesized nanoparticles of Odontosoria chinensis could be tested as an alternative to anticancer, anti-diabetic and anti-inflammatory drugs. Full article

(This article belongs to the Special Issue New Tools for Medicinal Chemists)

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8 pages, 654 KiB

Open AccessOpinion

COVID-19: A Brief Overview of the Discovery Clinical Trial

by Jean Jacques Vanden Eynde

Pharmaceuticals 2020, 13(4), 65; https://doi.org/10.3390/ph13040065 - 10 Apr 2020

Cited by 30 | Viewed by 10931

Abstract

The outbreak of COVID-19 is leading to a tremendous search for curative treatments. The urgency of the situation favors a repurposing of active drugs but not only antivirals. This short communication focuses on four treatments recommended by WHO and included in the first [...] Read more.

The outbreak of COVID-19 is leading to a tremendous search for curative treatments. The urgency of the situation favors a repurposing of active drugs but not only antivirals. This short communication focuses on four treatments recommended by WHO and included in the first clinical trial of the European Discovery project. Full article

(This article belongs to the Special Issue COVID-19 in Pharmaceuticals)

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17 pages, 2194 KiB

Open AccessArticle

Oleacein and Foam Cell Formation in Human Monocyte-Derived Macrophages: A Potential Strategy against Early and Advanced Atherosclerotic Lesions

by Agnieszka Filipek, Tomasz P. Mikołajczyk, Tomasz J. Guzik and Marek Naruszewicz

Pharmaceuticals 2020, 13(4), 64; https://doi.org/10.3390/ph13040064 - 9 Apr 2020

Cited by 22 | Viewed by 4277

Abstract

Background: Oleacein is a secoiridoid group polyphenol found mostly in Olea europea L. and Ligustrum vulgare L. (Oleaceae). The aim of the present study was to investigate a potential role of oleacein in prevention of the foam cell formation. Materials and Methods: Oleacein [...] Read more.

Background: Oleacein is a secoiridoid group polyphenol found mostly in Olea europea L. and Ligustrum vulgare L. (Oleaceae). The aim of the present study was to investigate a potential role of oleacein in prevention of the foam cell formation. Materials and Methods: Oleacein was isolated from Ligustrum vulgare leaves. Human monocyte-derived macrophages were obtained from monocytes cultured with Granulocyte-macrophage colony-stimulating factor (GM-CSF). Then, cells were incubated with 20 μM or 50 μM of oleacein and with oxidized low-density lipoprotein (oxLDL) (50 μg/mL). Visualization of lipid deposition within macrophages was carried out using Oil-Red-O. Expression of CD36, Scavenger receptor A1 (SRA1) and Lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1) was determined by Reverse transcription polymerase chain reaction (RT-PCR) and by flow cytometry. Apoptosis was determined by flow cytometry using Annexin V assay. STAT3 and Acyl-coenzyme A: cholesterol acyltransferase type 1 (ACAT1) levels were determined by ELISA. P-STAT3, P-JAK1, P-JAK2 expressions were determined by Western blot (WB). Results: Oleacein in dose-dependent manner significantly reduced lipid deposits in macrophages as well as their expression of selected scavenger receptors. The highest decrease of expression was found for CD36 and SRA1 receptors, from above 20% to more than 75% compared to oxLDL and the lowest for LOX-1 receptor, from approx. 8% to approx. 25% compared to oxLDL-stimulated macrophages. Oleacein significantly reduced (2.5-fold) early apoptosis of oxLDL-stimulated macrophages. Moreover, oleacein significantly increased the protein expression of JAK/STAT3 pathway and had no effect on ACAT1 level. Conclusions: Our study demonstrates, for the first time, that oleacein inhibits foam cell formation in human monocyte-derived macrophages and thus can be a valuable tool in the prevention of early and advanced atherosclerotic lesions. Full article

(This article belongs to the Section Natural Products)

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Graphical abstract

Graphical abstract
Full article ">Figure 1
Chemical structure of oleacein. Full article ">Figure 2
Influence of oleacein on oxLDL-induced foam cell formation in macrophages. The macrophages were incubated with or without oleacein (20 μM and 50 μM) or pitavatatin for 1 h followed by incubation with oxLDL at a concentration of 50 μg/mL for 72 h. (A) Oil Red O staining was used to visualize lipid deposition in macrophages (n = 12). (B) Data is a percentage of cells with lipid deposition compared to the oxLDL-induced macrophages (100%). Statistical significance * p < 0.05, ** p < 0.001. #—constant relative to which statistics are calculated. Full article ">Figure 3
(A) Influence of oleacein on CD36 mRNA, SRA1 mRNA and LOX-1 mRNA expression. mRNA levels are shown as arbitrary units normalized to GAPDH expression. Data from 15 experiments ± SEM. Statistical significance * p < 0.05, ** p < 0.001 compared to the oxLDL-induced macrophages. (B) Influence of oleacein on CD36, SRA1 and LOX-1 expression. The results are presented as the number of cells with CD36, SRA1 or LOX-1 expression ± SEM (n = 15). Statistical significance * p < 0.05, ** p < 0.001 compared to the oxLDL-induced macrophages. In dot plots representative results are shown. #—constant relative to which statistics are calculated. Full article ">Figure 4
Influence of oleacein on oxLDL-induced early apoptosis. The macrophages were incubated with oxLDL (100 μg/mL) or oxLDL with oleacein (20 μM and 50 μM) for 24 h. (A) Percentage of early apoptotic cells (n = 12) compared to the oxLDL-induced macrophages. Statistical significance ** p < 0.001. #—constant relative to which statistics are calculated. (B) Representative results from one of the twelve experiments are shown. Early apoptosis cells with Annexin V+/PI− (Q4). Full article ">Figure 5
(A) Influence of oleacein on STAT3 cellular level by oxLDL-induced macrophages was analysed by ELISA test. The experiment was performed on cells from different donors (n = 15). Statistical significance * p < 0.05, ** p < 0.001 compared oxLDL-induced macrophages. (B) Phosphorylation of STAT3 (P-STAT3/STAT3), JAK1 (P-JAK1/JAK) and JAK2 (P-JAK2/JAK2) was analysed by Western Blot and based on total protein. The results were quantified by densitometry. Representative images from one of tree experiments are shown. β-actin was used as an internal control. Vehicle: statistical significance ** p < 0.001 compared to the oxLDL-induced macrophages. As an inhibitor of JAK/STAT3 pathway. AZD1480, was used. Statistical significance ** p < 0.001 compared to factors without AZD1480. Full article ">

12 pages, 835 KiB

Open AccessArticle

Study of the Potential of the Capsule Shell Based on Natural Polysaccharides in Targeted Delivery of the L-Phenylalanine Ammonia-Lyase Enzyme Preparation

by Olga Babich, Lyubov Dyshlyuk, Alexander Prosekov, Svetlana Noskova, Oksana Ivina, Valery Pavsky, Svetlana Ivanova and Olga Bulgakova

Pharmaceuticals 2020, 13(4), 63; https://doi.org/10.3390/ph13040063 - 9 Apr 2020

Cited by 7 | Viewed by 3899

Abstract

The treatment of classical phenylketonuria is currently represented by many new methods of disease management. A promising method is the use of the enzyme L-phenylalanine ammonia-lyase (PAL) in various forms. The widespread use of enzyme preparations in therapy is limited by a lack [...] Read more.

The treatment of classical phenylketonuria is currently represented by many new methods of disease management. A promising method is the use of the enzyme L-phenylalanine ammonia-lyase (PAL) in various forms. The widespread use of enzyme preparations in therapy is limited by a lack of understanding of the mechanisms and systems of the targeted transport of PAL into certain organs and tissues as a result of the incorporation of a drug into the carrier. To ensure the stability of enzymes during the delivery process, encapsulation is preferable, which, as a rule, ensures the preservation of the qualitative characteristics of the enzymes orally applied to the environmental effects of the gastrointestinal tract (acidity, temperature, oxidation, etc.). Capsule preparations showed sufficient stability in the model gastric fluids and sustained release of the drug in the simulated intestinal fluid. Currently, there is a wide range of polymers used for encapsulation. The use of natural sources in the production technology of capsule systems improves bioavailability, controls the release, and prolongs the half-life of active substances. The advantage of this method is that the used enzyme is completely protected by the cell membranes of the capsules, which preserve its stability in the aggressive environment of the gastrointestinal tract. Capsules were obtained on the basis of compositions of hydrocolloids of plant origin. The potential of the developed capsules for targeted delivery of the enzyme preparation was studied. The degradation of the encapsulated form of the PAL enzyme preparation was studied in vitro in model bio-relevant media simulating the gastric and intestinal environment. The dynamics of the breakdown of the capsule shell allow us to expect that the release of L-phenylalanine ammonia-lyase from capsules based on plant hydrocolloids will occur no earlier than reaching the upper intestines, where the interaction with the protein components of the consumed food products to neutralize phenylalanine should occur. Full article

(This article belongs to the Section Pharmaceutical Technology)

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32 pages, 547 KiB

Open AccessReview

Inhibition of Fast Nerve Conduction Produced by Analgesics and Analgesic Adjuvants—Possible Involvement in Pain Alleviation

by Eiichi Kumamoto

Pharmaceuticals 2020, 13(4), 62; https://doi.org/10.3390/ph13040062 - 5 Apr 2020

Cited by 6 | Viewed by 4136

Abstract

Nociceptive information is transmitted from the periphery to the cerebral cortex mainly by action potential (AP) conduction in nerve fibers and chemical transmission at synapses. Although this nociceptive transmission is largely inhibited at synapses by analgesics and their adjuvants, it is possible that [...] Read more.

Nociceptive information is transmitted from the periphery to the cerebral cortex mainly by action potential (AP) conduction in nerve fibers and chemical transmission at synapses. Although this nociceptive transmission is largely inhibited at synapses by analgesics and their adjuvants, it is possible that the antinociceptive drugs inhibit nerve AP conduction, contributing to their antinociceptive effects. Many of the drugs are reported to inhibit the nerve conduction of AP and voltage-gated Na⁺ and K⁺ channels involved in its production. Compound action potential (CAP) is a useful measure to know whether drugs act on nerve AP conduction. Clinically-used analgesics and analgesic adjuvants (opioids, non-steroidal anti-inflammatory drugs, α₂-adrenoceptor agonists, antiepileptics, antidepressants and local anesthetics) were found to inhibit fast-conducting CAPs recorded from the frog sciatic nerve by using the air-gap method. Similar actions were produced by antinociceptive plant-derived chemicals. Their inhibitory actions depended on the concentrations and chemical structures of the drugs. This review article will mention the inhibitory actions of the antinociceptive compounds on CAPs in frog and mammalian peripheral (particularly, sciatic) nerves and on voltage-gated Na⁺ and K⁺ channels involved in AP production. Nerve AP conduction inhibition produced by analgesics and analgesic adjuvants is suggested to contribute to at least a part of their antinociceptive effects. Full article

14 pages, 1840 KiB

Open AccessArticle

Trends in Antidepressants Use in Spain between 2015 and 2018: Analyses from a Population-Based Registry Study with Reference to Driving

by Eduardo Gutiérrez-Abejón, Francisco Herrera-Gómez, Paloma Criado-Espegel and F. Javier Álvarez

Pharmaceuticals 2020, 13(4), 61; https://doi.org/10.3390/ph13040061 - 3 Apr 2020

Cited by 10 | Viewed by 4048

Abstract

Antidepressants are considered driving-impairing medicines (DIM). This is a population-based registry study that shows the trend in the use of antidepressants in Castile and León, Spain, from 2015 to 2018. Data on antidepressant dispensations at pharmacies and the adjusted use of these medicines [...] Read more.

Antidepressants are considered driving-impairing medicines (DIM). This is a population-based registry study that shows the trend in the use of antidepressants in Castile and León, Spain, from 2015 to 2018. Data on antidepressant dispensations at pharmacies and the adjusted use of these medicines by the driver population are presented. For the purposes of analysis, population distribution by age and gender has been taken into account, as well as the three Driving Under the Influence of Drugs, alcohol, and medicines (DRUID) categories. Antidepressants were used by 8.56% of the general population and 5.66% of drivers. Antidepressants were used more commonly by females than by males (12.12% vs. 4.87%, χ² = 1325.124, p = 0.001), and users increased as the age increased, even if women who drive used less antidepressants after turning 60 years of age. Chronic use of antidepressants was relevant (8.28%) in the same way as daily use (3.15%). Most of the consumption included SSRIs (4.99%), which are also known as “other antidepressants” (3.71%). Regardless of antidepressants consumed, users took 2.75 ± 1.19 DIMs, which are mainly anxiolytics (58.80%) and opioids (26.43%). Lastly, regarding consumption of antidepressants according to the DRUID classification, category I predominated over categories II and III. Our findings should serve as a starting point for health and traffic authorities to raise awareness of the risk for traffic accidents, especially involving SSRIs. Full article

(This article belongs to the Special Issue Choices of the Journal)

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17 pages, 1306 KiB

Open AccessReview

The Treatment of Impaired Wound Healing in Diabetes: Looking among Old Drugs

by Simona Federica Spampinato, Grazia Ilaria Caruso, Rocco De Pasquale, Maria Angela Sortino and Sara Merlo

Pharmaceuticals 2020, 13(4), 60; https://doi.org/10.3390/ph13040060 - 1 Apr 2020

Cited by 239 | Viewed by 21531

Abstract

Chronic wounds often occur in patients with diabetes mellitus due to the impairment of wound healing. This has negative consequences for both the patient and the medical system and considering the growing prevalence of diabetes, it will be a significant medical, social, and [...] Read more.

Chronic wounds often occur in patients with diabetes mellitus due to the impairment of wound healing. This has negative consequences for both the patient and the medical system and considering the growing prevalence of diabetes, it will be a significant medical, social, and economic burden in the near future. Hence, the need for therapeutic alternatives to the current available treatments that, although various, do not guarantee a rapid and definite reparative process, appears necessary. We here analyzed current treatments for wound healing, but mainly focused the attention on few classes of drugs that are already in the market with different indications, but that have shown in preclinical and few clinical trials the potentiality to be used in the treatment of impaired wound healing. In particular, repurposing of the antiglycemic agents dipeptidylpeptidase 4 (DPP4) inhibitors and metformin, but also, statins and phenyotin have been analyzed. All show encouraging results in the treatment of chronic wounds, but additional, well designed studies are needed to allow these drugs access to the clinics in the therapy of impaired wound healing. Full article

(This article belongs to the Collection Old Pharmaceuticals with New Applications)

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14 pages, 1661 KiB

Open AccessArticle

In Vitro Antibacterial and Antiproliferative Potential of Echinops lanceolatus Mattf. (Asteraceae) and Identification of Potential Bioactive Compounds

by Armel Jackson Seukep, Yong-Li Zhang, Yong-Bing Xu and Ming-Quan Guo

Pharmaceuticals 2020, 13(4), 59; https://doi.org/10.3390/ph13040059 - 30 Mar 2020

Cited by 30 | Viewed by 4607

Abstract

Many species belonging to the genus Echinops are widely used in traditional medicine to treat infectious diseases and cancers. The present study aimed to evaluate the antibacterial and antiproliferative properties of Echinops lanceolatus Mattf. (Asteraceae). The activity of the methanolic extract and subsequent [...] Read more.

Many species belonging to the genus Echinops are widely used in traditional medicine to treat infectious diseases and cancers. The present study aimed to evaluate the antibacterial and antiproliferative properties of Echinops lanceolatus Mattf. (Asteraceae). The activity of the methanolic extract and subsequent partition fractions was investigated against drug-resistant bacteria (Gram-negative and Gram-positive) and human tumor cell lines using broth microdilution and sulforhodamine B (SRB) assay, respectively. Our findings revealed weak to moderate antibacterial activities of tested extracts, with the recorded minimal inhibitory concentrations ranging from 256 to 1024 µg/mL. The ethyl acetate fraction (EL-EA) was found to be the most effective. Likewise, that fraction displayed strong antiproliferative potential with recorded IC₅₀ of 8.27 µg/mL and 28.27 µg/mL on A549 and HeLa cells, respectively. An analysis based on the ultra-performance liquid chromatography–electrospray ionization tandem mass spectrometry (UPLC–ESI–MS/MS) of the EL-EA fraction allowed the identification of 32 compounds, of which quinic acid and derivatives, cinnamic acid derivatives, dihydrokaempferol, naringenin-7-O-glucoside, apigenin-7-O-d-glucoside, naringin, apigenin, rhoifolin, coniferyl aldehyde, and secoisolariciresinol are well-known compounds of biological importance. This study is first to report on the biological activity and phytochemical profile of E. lanceolatus. We provide a baseline to consider E. lanceolatus as a valuable source of anti-infective and antiproliferative agents. Full article

(This article belongs to the Special Issue Bioactive Compounds from Plants and Foods with Pharmaceutical Interest)

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23 pages, 3509 KiB

Open AccessArticle

Upregulated Connexin 43 Induced by Loss-of-Functional S284L-Mutant α4 Subunit of Nicotinic ACh Receptor Contributes to Pathomechanisms of Autosomal Dominant Sleep-Related Hypermotor Epilepsy

by Kouji Fukuyama, Masashi Fukuzawa, Ruri Okubo and Motohiro Okada

Pharmaceuticals 2020, 13(4), 58; https://doi.org/10.3390/ph13040058 - 29 Mar 2020

Cited by 26 | Viewed by 3917

Abstract

To study the pathomechanism and pathophysiology of autosomal dominant sleep-related hypermotor epilepsy (ADSHE), this study determined functional abnormalities of glutamatergic transmission in the thalamocortical motor pathway, from the reticular thalamic nucleus (RTN), motor thalamic nuclei (MoTN) tosecondary motor cortex (M2C) associated with the [...] Read more.

To study the pathomechanism and pathophysiology of autosomal dominant sleep-related hypermotor epilepsy (ADSHE), this study determined functional abnormalities of glutamatergic transmission in the thalamocortical motor pathway, from the reticular thalamic nucleus (RTN), motor thalamic nuclei (MoTN) tosecondary motor cortex (M2C) associated with the S286L-mutant α4β2-nicotinic acetylcholine receptor (nAChR) and the connexin43 (Cx43) hemichannel of transgenic rats bearing the rat S286L-mutant Chrna4 gene (S286L-TG), which corresponds to the human S284L-mutant CHRNA4 gene using multiprobe microdialysis, primary cultured astrocytes and a Simple Western system. Expression of Cx43 in the M2C plasma membrane fraction of S286L-TG was upregulated compared with wild-type rats. Subchronic nicotine administration decreased Cx43 expression of wild-type, but did not affect that of S286L-TG; however, zonisamide (ZNS) decreased Cx43 in both wild-type and S286L-TG. Primary cultured astrocytes of wild-type were not affected by subchronic administration of nicotine but was decreased by ZNS. Upregulated Cx43 enhanced glutamatergic transmission during both resting and hyperexcitable stages in S286L-TG. Furthermore, activation of glutamatergic transmission associated with upregulated Cx43 reinforced the prolonged Cx43 hemichannel activation. Subchronic administration of therapeutic-relevant doses of ZNS compensated the upregulation of Cx43 and prolonged reinforced activation of Cx43 hemichannel induced by physiological hyperexcitability during the non-rapid eye movement phase of sleep. The present results support the primary pathomechanisms and secondary pathophysiology of ADSHE seizures of patients with S284L-mutation. Full article

(This article belongs to the Special Issue Therapeutic Agents for Neurological Disorders)

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26 pages, 2950 KiB

Open AccessArticle

Efficacy of Panax ginseng Meyer Herbal Preparation HRG80 in Preventing and Mitigating Stress-Induced Failure of Cognitive Functions in Healthy Subjects: A Pilot, Randomized, Double-Blind, Placebo-Controlled Crossover Trial

by Pierre-Antoine Mariage, Areg Hovhannisyan and Alexander G. Panossian

Pharmaceuticals 2020, 13(4), 57; https://doi.org/10.3390/ph13040057 - 29 Mar 2020

Cited by 14 | Viewed by 6153

Abstract

Background: The aim of this pilot study was to compare the efficacy of hydroponically cultivated red Panax ginseng Meyer root preparation (HRG80) and traditionally harvested six-year-old white P. ginseng standard preparation (PGS) with placebo in preventing symptoms of stress. Methods: The effects of [...] Read more.

Background: The aim of this pilot study was to compare the efficacy of hydroponically cultivated red Panax ginseng Meyer root preparation (HRG80) and traditionally harvested six-year-old white P. ginseng standard preparation (PGS) with placebo in preventing symptoms of stress. Methods: The effects of HRG80, PGS, and placebo capsules were studied in 50 tired healthy subjects in a three-arm, randomized, double-blinded, placebo-controlled crossover trial. Efficacy-outcome measures included the accuracy of processing the d2 test for cognitive functions, obtained accuracy score in a computerized memory test, and the perceived-stress (PS) score. Results: A statistically significant interaction effect between time and treatment (p < 0.0001) was observed in the attention d2 and memory tests, indicating that HRG80 treatment was more beneficial than that with a placebo. The effects of PGS were better than those of the placebo, but the difference was not statistically significant. There was significant difference between the effects of HRG80 and PGS (p < 0.0001) that were observed after single (Day 1) and repeated administrations on Days 5 and 12 of treatment. Conclusion: Overall, HRG80 treatment was significantly superior compared to that with the PGS and placebo regarding attention, memory, and PS scores after single and repeated administrations for 5 and 12 days. Full article

(This article belongs to the Special Issue Medicinal Plants 2020)

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12 pages, 2063 KiB

Open AccessArticle

Selection of Thai Medicinal Plants with Anti-Obesogenic Potential via In Vitro Methods

by Wijitrapha Ruangaram and Eisuke Kato

Pharmaceuticals 2020, 13(4), 56; https://doi.org/10.3390/ph13040056 - 29 Mar 2020

Cited by 7 | Viewed by 4504

Abstract

The prevalence of obesity is increasing globally. Despite the availability of a variety of anti-obesogenic drugs, including therapies under clinical development, these treatments are often indicated for patients with severe obesity, making them unsuitable for patients with mild obesity or for preventative use. [...] Read more.

The prevalence of obesity is increasing globally. Despite the availability of a variety of anti-obesogenic drugs, including therapies under clinical development, these treatments are often indicated for patients with severe obesity, making them unsuitable for patients with mild obesity or for preventative use. In Thailand, traditional remedies employing medicinal plants are widely used to maintain health and treat disease. These treatments are generally inexpensive and readily available at markets, making them good treatment options for preventing obesity. To evaluate the anti-obesogenic potential of Thai medicinal plants, we employed three in vitro methods: pancreatic lipase inhibition, lipolysis enhancement, and lipid accumulation reduction assays. Among 70 Thai medicinal plants, Eurycoma longifolia Jack, Tiliacora triandra Diels, and Acacia concinna (Willd.) DC. were selected as the most favorable candidates because they exhibited anti-obesogenic activity in all three assays. These medicinal plants are expected to have efficient anti-obesogenic effects, making them promising candidates for further study. Full article

(This article belongs to the Special Issue Medicinal Plants 2020)

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Pancreatic lipase-inhibitory activities of the candidate plant extracts. Cetilistat (positive control) concentration was 8 µM. Data are expressed as means ± standard deviation. Means with different letters are significantly different (Tukey’s test, p < 0.05). Full article ">Figure 2
Lipolysis enhancement activity of the candidate plant extracts. Isoproterenol (ISP, positive control) was used at 2.5 µM. Data are expressed as means ± standard deviation. Means with different letters are significantly different (Tukey’s test, p < 0.05). Full article ">Figure 3
Lipolysis enhancement activity of the Acacia concinna extract. Data are expressed as means ± standard deviation. Means with different letters are significantly different (Tukey’s test, p < 0.05). Full article ">Figure 4
Lipid accumulation reduction activities of the candidate plant extracts. Brucea javanica (positive control) was used at 0.05 mg/mL. Data are expressed as means ± standard deviation. Means with different letters are significantly different (Tukey’s test, p < 0.05). Full article ">Figure 5
Cell viability after treatment with the candidate plant extracts. Triton X-100 (positive control) was used at 0.1% v/v. Data are expressed as means ± standard deviation. Means with different letters are significantly different (Tukey’s test, p < 0.05). Full article ">

12 pages, 842 KiB

Open AccessArticle

Antioxidant, Anti-inflammatory Activities and Polyphenol Profile of Rhamnus prinoides

by Gui-Lin Chen, Fredrick Munyao Mutie, Yong-Bing Xu, Flora Didii Saleri, Guang-Wan Hu and Ming-Quan Guo

Pharmaceuticals 2020, 13(4), 55; https://doi.org/10.3390/ph13040055 - 26 Mar 2020

Cited by 35 | Viewed by 7217

Abstract

Rhamnus prinoides L’Herit (R. prinoides) has long been widely consumed as folk medicine in Kenya and other Africa countries. Previous studies indicated that polyphenols were abundant in genus Rhamnus and exhibited outstanding antioxidant and anti-inflammatory activities. However, there are very few [...] Read more.

Rhamnus prinoides L’Herit (R. prinoides) has long been widely consumed as folk medicine in Kenya and other Africa countries. Previous studies indicated that polyphenols were abundant in genus Rhamnus and exhibited outstanding antioxidant and anti-inflammatory activities. However, there are very few studies on such pharmacological activities and the polyphenol profile of this plant up to now. In the present study, the antioxidant activities of the crude R. prinoides extracts (CRE) and the semi-purified R. prinoides extracts (SPRE) of polyphenol enriched fractions were evaluated to show the strong radical scavenging effects against 1,1-diphenyl-2- picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl (DPPH) (0.510 ± 0.046 and 0.204 ± 0.005, mg/mL), and 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) (0.596 ± 0.005 and 0.096 ± 0.004, mg/mL), respectively. Later, the SPRE with higher contents of polyphenols and flavonoids displayed obvious anti-inflammatory activities through reducing the NO production at the dosage of 11.11 − 100 μg/mL, and the COX-2 inhibitory activity with an IC₅₀ value at 20.61 ± 0.13 μg/mL. Meanwhile, the HPLC-UV/ESI-MS/MS analysis of polyphenol profile of R. prinoides revealed that flavonoids and their glycosides were the major ingredients, and potentially responsible for its strong antioxidant and anti-inflammatory activities. For the first time, the present study comprehensively demonstrated the chemical profile of R. prinoides, as well as noteworthy antioxidant and anti-inflammatory activities, which confirmed that R. prinoides is a good natural source of polyphenols and flavonoids, and provided valuable information on this medicinal plant as folk medicine and with good potential for future healthcare practice. Full article

(This article belongs to the Special Issue Medicinal Plants 2020)

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18 pages, 3390 KiB

Open AccessArticle

Improved Surface Display of Human Hyal1 and Identification of Testosterone Propionate and Chicoric Acid as New Inhibitors

by Isabelle Lengers, Fabian Herrmann, Marc Le Borgne and Joachim Jose

Pharmaceuticals 2020, 13(4), 54; https://doi.org/10.3390/ph13040054 - 26 Mar 2020

Cited by 8 | Viewed by 3743

Abstract

Degradation of high molecular weight hyaluronic acid (HA) in humans is mainly catalyzed by hyaluronidase Hyal1. This enzyme is involved in many pathophysiological processes and therefore appears an interesting target for drug discovery. Until now, only a few inhibitors of human Hyal1 are [...] Read more.

Degradation of high molecular weight hyaluronic acid (HA) in humans is mainly catalyzed by hyaluronidase Hyal1. This enzyme is involved in many pathophysiological processes and therefore appears an interesting target for drug discovery. Until now, only a few inhibitors of human Hyal1 are known due to obstacles in obtaining active enzymes for inhibitor screening. The aim of the present work was to provide a convenient enzyme activity assay and show its feasibility by the identification of new inhibitors. By autodisplay, Escherichia coli F470 can present active Hyal1 on its surface. In this study, the inducible expression of Hyal1 on the cell surface of E. coli under the control of a rhamnose-dependent promoter (P_rha) was performed and optimized. Enzyme activity per single cell was increased by a factor of 100 compared to the constitutive Hyal1 surface display, as described before. An activity of 6.8 × 10⁻⁴ mU per single cell was obtained under optimal reaction conditions. By this modified activity assay, two new inhibitors of human Hyal1 were identified. Chicoric acid, a natural compound belonging to the phenylpropanoids, showed an IC₅₀ value of 171 µM. The steroid derivative testosterone propionate showed and IC₅₀ value of 124 ± 1.1 µM. Both values were in the same order of magnitude as the IC₅₀ value of glycyrrhizic acid (177 µM), one of the best known inhibitors of human Hyal1 known so far. In conclusion, we established a new enzyme activity assay for human Hyal1 and identified new inhibitors with this new assay method. Full article

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14 pages, 2773 KiB

Open AccessArticle

Study of In Vitro and In Vivo Carbamazepine Release from Coarse and Nanometric Pharmaceutical Emulsions Obtained via Ultra-High-Pressure Homogenization

by Juan D. Echeverri, Maria J. Alhajj, Nicolle Montero, Cristhian J. Yarce, Alvaro Barrera-Ocampo and Constain H. Salamanca

Pharmaceuticals 2020, 13(4), 53; https://doi.org/10.3390/ph13040053 - 26 Mar 2020

Cited by 8 | Viewed by 4287

Abstract

In the past decade, pharmaceutical nanotechnology has proven to be a promising alternative for improving the physicochemical and biopharmaceutical features for conventional pharmaceutical drug formulations. The goal of this study was to develop, characterize, and evaluate the in vitro and in vivo release [...] Read more.

In the past decade, pharmaceutical nanotechnology has proven to be a promising alternative for improving the physicochemical and biopharmaceutical features for conventional pharmaceutical drug formulations. The goal of this study was to develop, characterize, and evaluate the in vitro and in vivo release of the model drug carbamazepine (CBZ) from two emulsified formulations with different droplet sizes (coarse and nanometric). Briefly, oil-in-water emulsions were developed using (i) Sacha inchi oil, ultrapure water, Tween^TM 80, and Span^TM 80 as surfactants, (ii) methyl-paraben and propyl-paraben as preservatives, and (iii) CBZ as a nonpolar model drug. The coarse and nanometric emulsions were prepared by rotor–stator dispersion and ultra-high-pressure homogenization (UHPH), respectively. The in vitro drug release studies were conducted by dialysis, whereas the in vivo drug release was evaluated in New Zealand breed rabbits. The results showed that nanoemulsions were physically more stable than coarse emulsions, and that CBZ had a very low release for in vitro determination (<2%), and a release of 20% in the in vivo study. However, it was found that nanoemulsions could significantly increase drug absorption time from 12 h to 45 min. Full article

(This article belongs to the Section Pharmaceutical Technology)

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