Pharmaceuticals

15 pages, 1275 KiB

Open AccessArticle

The Probiotic VSL#3^® Does Not Seem to Be Efficacious for the Treatment of Gastrointestinal Symptomatology of Patients with Fibromyalgia: A Randomized, Double-Blind, Placebo-Controlled Clinical Trial

by Elena P. Calandre, Javier Hidalgo-Tallon, Rocio Molina-Barea, Fernando Rico-Villademoros, Cristina Molina-Hidalgo, Juan M. Garcia-Leiva, Maria Dolores Carrillo-Izquierdo and Mahmoud Slim

Pharmaceuticals 2021, 14(10), 1063; https://doi.org/10.3390/ph14101063 - 19 Oct 2021

Cited by 7 | Viewed by 4699

Abstract

Gastrointestinal symptomatology is frequent among patients with fibromyalgia, which increases disease burden and lacks specific treatment, either pharmacological or non-pharmacological. We aimed to evaluate the efficacy and tolerability of a multi-strain probiotic, VSL#3^®, for the treatment of fibromyalgia-associated gastrointestinal manifestations. This [...] Read more.

Gastrointestinal symptomatology is frequent among patients with fibromyalgia, which increases disease burden and lacks specific treatment, either pharmacological or non-pharmacological. We aimed to evaluate the efficacy and tolerability of a multi-strain probiotic, VSL#3^®, for the treatment of fibromyalgia-associated gastrointestinal manifestations. This randomized, placebo-controlled trial included 12 weeks of probiotic or placebo treatment followed by 12 weeks of follow up. The primary outcome variable was the mean change from the baseline to the endpoint in the composite severity score of the three main gastrointestinal symptoms reported by patients with fibromyalgia (abdominal pain, abdominal bloating and meteorism). Secondary outcome variables were the severity of additional gastrointestinal symptoms, fibromyalgia severity, depression, sleep disturbance, health-related quality of life and patients’ overall impression of improvement. No differences were found between VSL#3^® (n = 54) and the placebo (n = 56) in the primary outcome (estimated treatment difference: 1.1; 95% confidence interval [CI]: ?2.1, 4.2; p = 0.501), or in any of the secondary outcomes. However, responders to VSL#3 were more likely to maintain any improvement during the follow-up period compared to responders in the placebo arm. Overall, VSL#3 tolerability was good. Our data could not demonstrate any beneficial effects of VSL#3^® either on the composite score of severity of abdominal pain, bloating and meteorism or in any of the secondary outcome variables. More research is needed to elucidate specific factors that may predict a favourable response to treatment in patients with fibromyalgia. Full article

(This article belongs to the Special Issue Rheumatic Diseases: Pathophysiology, Targeted Therapy, Focus on Vascular and Pulmonary Manifestations)

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99 pages, 23166 KiB

Open AccessReview

Mechanistic Understanding from Molecular Dynamics in Pharmaceutical Research 2: Lipid Membrane in Drug Design

by Tomasz Róg, Mykhailo Girych and Alex Bunker

Pharmaceuticals 2021, 14(10), 1062; https://doi.org/10.3390/ph14101062 - 19 Oct 2021

Cited by 39 | Viewed by 15444

Abstract

We review the use of molecular dynamics (MD) simulation as a drug design tool in the context of the role that the lipid membrane can play in drug action, i.e., the interaction between candidate drug molecules and lipid membranes. In the standard “lock [...] Read more.

We review the use of molecular dynamics (MD) simulation as a drug design tool in the context of the role that the lipid membrane can play in drug action, i.e., the interaction between candidate drug molecules and lipid membranes. In the standard “lock and key” paradigm, only the interaction between the drug and a specific active site of a specific protein is considered; the environment in which the drug acts is, from a biophysical perspective, far more complex than this. The possible mechanisms though which a drug can be designed to tinker with physiological processes are significantly broader than merely fitting to a single active site of a single protein. In this paper, we focus on the role of the lipid membrane, arguably the most important element outside the proteins themselves, as a case study. We discuss work that has been carried out, using MD simulation, concerning the transfection of drugs through membranes that act as biological barriers in the path of the drugs, the behavior of drug molecules within membranes, how their collective behavior can affect the structure and properties of the membrane and, finally, the role lipid membranes, to which the vast majority of drug target proteins are associated, can play in mediating the interaction between drug and target protein. This review paper is the second in a two-part series covering MD simulation as a tool in pharmaceutical research; both are designed as pedagogical review papers aimed at both pharmaceutical scientists interested in exploring how the tool of MD simulation can be applied to their research and computational scientists interested in exploring the possibility of a pharmaceutical context for their research. Full article

(This article belongs to the Special Issue In Silico Approaches in Drug Design)

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11 pages, 436 KiB

Open AccessArticle

Poppers Use and High Methaemoglobinaemia: ‘Dangerous Liaisons’

by Malcolm Barrangou-Poueys-Darlas, Marie Gerardin, Sylvie Deheul, Marion Istvan, Marylène Guerlais, FAN, Pascale Jolliet, Thomas Dejoie and Caroline Victorri-Vigneau

Pharmaceuticals 2021, 14(10), 1061; https://doi.org/10.3390/ph14101061 - 19 Oct 2021

Cited by 6 | Viewed by 8486

Abstract

Poppers are legal and largely used in France despite severe side effects, such as methaemoglobinaemia (MetHbia). Our work aimed to assess the prevalence of poppers consumers among patients with a MetHbia higher than or equal to 5% in French university hospitals and its [...] Read more.

Poppers are legal and largely used in France despite severe side effects, such as methaemoglobinaemia (MetHbia). Our work aimed to assess the prevalence of poppers consumers among patients with a MetHbia higher than or equal to 5% in French university hospitals and its evolution before and after the legalization of poppers in France. We conducted a national multicentre observational retrospective study. All patients for whom at least one MetHbia measurement was performed from 2012 to 2017 in university hospitals where the French addictovigilance network (FAN) is implanted were included. For each MetHbia measurement exceeding or equal to 5%, a return to the clinical file was made by the FAN to assess poppers consumption. We calculated the prevalence of MetHbia exceeding or equal to 5% and 25% and the prevalence of poppers consumption before and after the legalization. A total of 239 (0.14%) patients had a MetHbia level exceeding or equal to 5% with 25 (10.46%) cases of poppers consumption. Poppers consumption represented 68.4% (13 out of 19) of cases with MetHbia greater than or equal to 25%. Poppers consumption among patients with MetHbia exceeding or equal to 5% increased after the legalization from 4.76% to 11.67% (prevalence ratio PR = 2.45, 95% CI = [0.98–8.37], p-value = 0.190). The proportion of patients with a MetHbia level of 25% or more increased after the legalization from 4.76% to 8.63% (PR = 1.81, 95% CI = [0.68–6.82], p-value = 0.374). The use of poppers is very frequently reported by patients with MetHbia greater than or equal to 25%. Full article

(This article belongs to the Special Issue Pharmacokinetics and Pharmacodynamics of Psychoactive Substances: Clinical and Forensic Aspects)

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18 pages, 2761 KiB

Open AccessArticle

Macrosphelide A Exhibits a Specific Anti-Cancer Effect by Simultaneously Inactivating ENO1, ALDOA, and FH

by Kyoung Song, Nirmal Rajasekaran, Chaithanya Chelakkot, Hun Seok Lee, Seung-Mann Paek, Hobin Yang, Lina Jia, Hee Geon Park, Woo Sung Son, Yu-Jin Kim, Joon-Seok Choi, Hae Min Jeong, Young-Ger Suh, Hwayoung Yun and Young Kee Shin

Pharmaceuticals 2021, 14(10), 1060; https://doi.org/10.3390/ph14101060 - 19 Oct 2021

Cited by 6 | Viewed by 4274

Abstract

Aerobic glycolysis in cancer cells, also known as the Warburg effect, is an indispensable hallmark of cancer. This metabolic adaptation of cancer cells makes them remarkably different from normal cells; thus, inhibiting aerobic glycolysis is an attractive strategy to specifically target tumor cells [...] Read more.

Aerobic glycolysis in cancer cells, also known as the Warburg effect, is an indispensable hallmark of cancer. This metabolic adaptation of cancer cells makes them remarkably different from normal cells; thus, inhibiting aerobic glycolysis is an attractive strategy to specifically target tumor cells while sparing normal cells. Macrosphelide A (MSPA), an organic small molecule, is a potential lead compound for the design of anti-cancer drugs. However, its role in modulating cancer metabolism remains poorly understood. MSPA target proteins were screened using mass spectrometry proteomics combined with affinity chromatography. Direct and specific interactions of MSPA with its candidate target proteins were confirmed by in vitro binding assays, competition assays, and simulation modeling. The siRNA-based knockdown of MSPA target proteins indirectly confirmed the cytotoxic effect of MSPA in HepG2 and MCF-7 cancer cells. In addition, we showed that MSPA treatment in the HEPG2 cell line significantly reduced glucose consumption and lactate release. MSPA also inhibited cancer cell proliferation and induced apoptosis by inhibiting critical enzymes involved in the Warburg effect: aldolase A (ALDOA), enolase 1 (ENO1), and fumarate hydratase (FH). Among these enzymes, the purified ENO1 inhibitory potency of MSPA was further confirmed to demonstrate the direct inhibition of enzyme activity to exclude indirect/secondary factors. In summary, MSPA exhibits anti-cancer effects by simultaneously targeting ENO1, ALDOA, and FH. Full article

(This article belongs to the Topic Compounds with Medicinal Value)

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(A) The cytotoxic effects of macrosphelide A (MSPA) in cancer cell lines and non-cancer cell lines at 48 h, 72 h, and 96 h time points. The mock group represents vehicle-treated cell lines. Data represent the mean ± SD (n = 3). * p < 0.05, ** p < 0.01 **** p < 0.0001 versus vehicle-treated group (mock). (B) Apoptosis was determined using annexin V/7-aminoactinomycin D (7-ADD) staining in HepG2 and HL60 cell lines. Combined annexin V and 7-AAD reactivity allowed cells to be classified into four groups: early apoptotic cells (annexin V (+) and 7-AAD (−)), late apoptotic or dead cells (annexin V (+) and 7-AAD (+)), dead cells (annexin V (−) and 7-AAD (+)), and live cells (annexin V (−) and 7-AAD (−)). Data represent the mean ± SD (n = 3). * p < 0.05, versus vehicle-treated cells. (C) HepG2 cells were treated with 500 µM of MSPA for 72 h and stained with annexin V Alexa Fluor 488 (green fluorescence). Cell nuclei are stained blue (Hoechst stain). Data represent the mean ± SD (n = 3). *** p < 0.001 versus control (mock). Full article ">Figure 2
(A) Analysis of the putative target proteins of MSPA by Coomassie-stained SDS-PAGE analysis. Bands submitted to in-gel tryptic digestion for further mass spectrometric analysis are marked with blue arrows (first arrow and second arrow). Standard molecular weight (kDa) is on the left. (B) Detection of the interaction between biotinylated MSPA (or butenylated biotin, MSPA biotin-961, or MSPA biotin-735) and aldolase A (ALDOA), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), enolase 1 (ENO1), and fumarate hydratase (FH) proteins. (C) Results of an in vitro binding assay to determine interactions between MSPA and ALDOA, GAPDH, ENO1, and FH; biotinylated MSPB was used as negative controls. (D) The binding specificity of MSPA in target proteins was confirmed using a competitive binding assay. Excess non-biotinylated MSPA (0× (12.5 µM), 100× (125 µM), and 1000× (1.25 mM)) was added to HL60 cell lysates as a competitor of biotinylated MSPA. Full article ">Figure 3
MSPA impairs glycolysis through modifying the enzymes (A), measurement of the effect of MSPA on the enzymatic activity of ALDOA, ENO1, FH, and GAPDH in HepG2 cell lines (B), relative lactate production (left) and (C), relative glucose consumption (right) in NT (DMSO-treated) and MSPA (50, 100, and 500 uM)-treated HepG2 cells for 72 h. (D), Extracellular acidification rate and (E), oxygen consumption rate in HepG2 cells treated with 50 µM, 100 µM and 500 µM of MSPA for 48 h. Data represent the mean ± SD (n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus the control group. Full article ">Figure 4
(A) The effect of siRNA-mediated gene knockdown on the expression level of ALDOA, ENO1, and FH mRNA. (B) The effect of ALDOA-, ENO1-, and FH-targeted siRNAs in HepG2 and MCF-7 cells. Lysates were prepared from cells pretreated with 1.2 nM of siRNA against a single-target enzyme. β-actin was used as a loading control. Total cellular proteins were separated using SDS-PAGE, transferred to PVDF membranes, and detected using specific antibodies against ALDOA, ENO1, FH, and β-actin. (C) The viability of HepG2 and MCF-7 cells (expressed as a percentage of the control) was determined using a CellTiter Glo luminescent assay kit. The cells were treated for 72 h with 1.2 nM of single-target siRNA (ALDOA, ENO1, or FH) and 1.2 nM of siRNA against more than one target (a combination of ALDOA, ENO1, and FH). Data were obtained from three independent experiments and presented as mean ± SD., **** p < 0.0001 versus the negative control siRNA (NC) group. Full article ">Figure 5
(A) Molecular docking calculations of MSPA–protein complexes (GAPDH (green), FH (brown), ALDOA (red), and ENO1 (blue)). The graph depicts total binding energy plotted against time. (B) Structural representation of molecular docking analysis of MSPA in complex with the human ENO1 protein. Models of complexed ENO1 proteins are represented as electrostatic surface potentials (ESPs) using the ball and stick method. Positively charged surface regions are colored blue, and negatively charged surface regions are colored red. Heavy atoms are colored yellow (for sulfur), red (for oxygen), and blue (for nitrogen) in the ball and stick model of the binding site. The MSPA molecule is colored green, and interactions between proteins are represented with cyan lines (for hydrogen bonds), yellow dots (for salt bridges), and gray dots (for hydrophobic interactions). Full article ">Figure 6
A schematic representation of the molecular mechanisms of MSPA activity; the compound inhibits cancer cell proliferation by targeting ALDOA, ENO1, and FH. Full article ">

14 pages, 2725 KiB

Open AccessArticle

Physicochemical Characteristics and In Vitro Toxicity/Anti-SARS-CoV-2 Activity of Favipiravir Solid Lipid Nanoparticles (SLNs)

by Alaa S. Tulbah and Wing-Hin Lee

Pharmaceuticals 2021, 14(10), 1059; https://doi.org/10.3390/ph14101059 - 19 Oct 2021

Cited by 25 | Viewed by 3872

Abstract

The rise of coronavirus (COVID-19) cases worldwide has driven the need to discover and develop novel therapeutics with superior efficacy to treat this disease. This study aims to develop an innovative aerosolized nano-formulation of favipiravir (FPV) as an anti-viral agent against coronavirus infection. [...] Read more.

The rise of coronavirus (COVID-19) cases worldwide has driven the need to discover and develop novel therapeutics with superior efficacy to treat this disease. This study aims to develop an innovative aerosolized nano-formulation of favipiravir (FPV) as an anti-viral agent against coronavirus infection. The local delivery of FPV nanoparticles (NPs) via nebulization ensures that the drug can reach the site of infection, the lungs. Solid lipid NPs of favipiravir (FPV-SLNs) were formulated utilizing the hot-evaporation method. The physicochemical formulation properties were evaluated using dynamic light scattering (DLS), Fourier-transform infrared spectroscopy (FTIR), and differential scanning calorimetry (DSC). The aerosol formulation performance was evaluated using an Andersen Cascade Impactor (ACI) at a flow rate of 15 L/min. The FPV-SLN formulation’s in vitro anti-viral activity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was also evaluated using the SARS-CoV-2 pathogen (hCoV-19/Egypt/NRC-3/2020 isolate). The FPV-SLNs’ morphology was defined utilizing transmission electron microscopy, showing an irregular shape. By means of FPV-SLNs’ nebulization, a fine particle fraction of 60.2 ± 1.7% was produced with 60.2 ± 1.7%, and this finding suggests that FPV-SLNs were appropriate for inhalation drug delivery with a particle size of 537.6 ± 55.72 nm. Importantly, the FPV-SLNs showed anti-viral activity against SARS-CoV-2 with CC₅₀ and IC50 values of 449.6 and 29.9 µg/mL, respectively. This study suggests that inhaled solid lipid NPs of favipiravir could potentially be used against coronavirus. Full article

(This article belongs to the Special Issue Emerging Platforms in Nanotechnology for Diagnosis and Treatment of Respiratory and Infectious Diseases)

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Transmission electron micrograph of FPV-SLN formulation. Full article ">Figure 2
In vitro drug release across a dialysis membrane of FPV-SLNs (n = 3, mean ± SD). Full article ">Figure 3
Drug release kinetics of FPV-SLNs fitted to mathematical kinetic model. (A) Zero order, (B) first order, (C) Higuchi model (D) Korsmeyer–Peppas model and (E) Hixson–Crowell model. It should be noted that only cumulative release of FPV until 2 h is fitted to the model. Solid black circle refers to FPV release. Full article ">Figure 4
FTIR spectra of (A) unprocessed FPV, (B) Compritol 888, (C) Tween 80, and (D) FPV-SLNs. Full article ">Figure 5
Differential scanning calorimetry thermographs of (A) unprocessed FPV, (B) Compritol 888, (C) Tween 80, and (D) FPV-SLNs. Full article ">Figure 6
Summarized data showing ACI different stages deposition of nebulized FPV-SLN formulations (n = 3, mean ± SD). Full article ">Figure 7
Determination of the antiviral activity of FPV-SLNs against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The dots represent the cell viability readouts at different treatment concentrations, and the squares represent the virus inhibition percentages of the treatment on Vero E6 cells. Data represent mean ± SD (n = 3). Full article ">

19 pages, 887 KiB

Open AccessReview

Zebrafish as a Model System to Study the Mechanism of Cutaneous Wound Healing and Drug Discovery: Advantages and Challenges

by Ruth Naomi, Hasnah Bahari, Muhammad Dain Yazid, Hashim Embong and Fezah Othman

Pharmaceuticals 2021, 14(10), 1058; https://doi.org/10.3390/ph14101058 - 18 Oct 2021

Cited by 28 | Viewed by 6685

Abstract

In humans, cutaneous wounds may heal without scars during embryogenesis. However, in the adult phase, the similar wound may undergo a few events such as homeostasis, blood clotting, inflammation, vascularization, and the formation of granulation tissue, which may leave a scar at the [...] Read more.

In humans, cutaneous wounds may heal without scars during embryogenesis. However, in the adult phase, the similar wound may undergo a few events such as homeostasis, blood clotting, inflammation, vascularization, and the formation of granulation tissue, which may leave a scar at the injury site. In consideration of this, research evolves daily to improve the healing mechanism in which the wound may heal without scarring. In regard to this, zebrafish (Danio rerio) serves as an ideal model to study the underlying signaling mechanism of wound healing. This is an important factor in determining a relevant drug formulation for wound healing. This review scrutinizes the biology of zebrafish and how this favors the cutaneous wound healing relevant to the in vivo evidence. This review aimed to provide the current insights on drug discovery for cutaneous wound healing based on the zebrafish model. The advantages and challenges in utilizing the zebrafish model for cutaneous wound healing are discussed in this review. This review is expected to provide an idea to formulate an appropriate drug for cutaneous wound healing relevant to the underlying signaling mechanism. Therefore, this narrative review recapitulates current evidence from in vivo studies on the cutaneous wound healing mechanism, which favours the discovery of new drugs. This article concludes with the need for zebrafish as an investigation model for biomedical research in the future to ensure that drug repositions are well suited for human skin. Full article

(This article belongs to the Special Issue Zebrafish as a Powerful Tool for Drug Discovery 2021)

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23 pages, 5080 KiB

Open AccessReview

Epilepsy in Neurodegenerative Diseases: Related Drugs and Molecular Pathways

by Amanda Cano, Elena Fonseca, Miren Ettcheto, Elena Sánchez-López, Itziar de Rojas, Silvia Alonso-Lana, Xavier Morató, Eliana B. Souto, Manuel Toledo, Mercè Boada, Marta Marquié and Agustín Ruíz

Pharmaceuticals 2021, 14(10), 1057; https://doi.org/10.3390/ph14101057 - 18 Oct 2021

Cited by 41 | Viewed by 11408

Abstract

Epilepsy is a chronic disease of the central nervous system characterized by an electrical imbalance in neurons. It is the second most prevalent neurological disease, with 50 million people affected around the world, and 30% of all epilepsies do not respond to available [...] Read more.

Epilepsy is a chronic disease of the central nervous system characterized by an electrical imbalance in neurons. It is the second most prevalent neurological disease, with 50 million people affected around the world, and 30% of all epilepsies do not respond to available treatments. Currently, the main hypothesis about the molecular processes that trigger epileptic seizures and promote the neurotoxic effects that lead to cell death focuses on the exacerbation of the glutamate pathway and the massive influx of Ca²⁺ into neurons by different factors. However, other mechanisms have been proposed, and most of them have also been described in other neurodegenerative diseases, such as Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, or multiple sclerosis. Interestingly, and mainly because of these common molecular links and the lack of effective treatments for these diseases, some antiseizure drugs have been investigated to evaluate their therapeutic potential in these pathologies. Therefore, in this review, we thoroughly investigate the common molecular pathways between epilepsy and the major neurodegenerative diseases, examine the incidence of epilepsy in these populations, and explore the use of current and innovative antiseizure drugs in the treatment of refractory epilepsy and other neurodegenerative diseases. Full article

(This article belongs to the Special Issue Epilepsy and Neurodegeneration: Current Therapeutic Implications 2021)

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General molecular mechanisms of the development of seizure activity in epilepsy and associated ASDs. Full article ">Figure 2
Seizure activity derived from the main pathological molecular pathways of Alzheimer’s disease. The pathological hallmarks of Alzheimer’s disease promote an increase in neuroinflammation and intracellular Ca2+ through ACh and NMDA receptors and Na+/Ca2+ channels. This promotes an increase in neuroinflammation and neuronal hyperexcitability, which in turn increases the neurodegeneration process (and vice versa) in a vicious cycle. NE, norepinephrine. Full article ">Figure 3
Related molecular pathways between Parkinson’s disease and epilepsy. (A) Neuronal excitability via mitochondrial dysfunction derived from the accumulation of abnormal α-synuclein. Abnormal α-synuclein promotes membrane depolarization, massive influx of intracellular Ca2+, and oxidative stress through the induction of mitochondrial dysfunction and Lewy bodies’ formation. This promotes an increase in neuroinflammation and neuronal hyperexcitability, which in turn increases the neurodegeneration process (and vice versa) in a vicious cycle. (B) Proepileptic/antiepileptic properties of dopamine conditioned by its binding to the D1/D2 family of receptors. Binding of dopamine to D1R promotes an increase in cAMP, which leads to the activation of NMDA-Rs and blockage of GLUT1, thus promoting a massive influx of intracellular Ca2+ and a reduction in glutamate reuptake. This gives rise to an increase in neuroinflammation and neuronal hyperexcitability, which in turn increases the neurodegeneration process (and vice versa) in a vicious cycle. Binding of dopamine to D2R inhibits the production of cAMP, thus promoting the opposite effect of that of D1R activation. NE, norepinephrine; ROS, reactive oxygen species. Full article ">Figure 3 Cont.
Related molecular pathways between Parkinson’s disease and epilepsy. (A) Neuronal excitability via mitochondrial dysfunction derived from the accumulation of abnormal α-synuclein. Abnormal α-synuclein promotes membrane depolarization, massive influx of intracellular Ca2+, and oxidative stress through the induction of mitochondrial dysfunction and Lewy bodies’ formation. This promotes an increase in neuroinflammation and neuronal hyperexcitability, which in turn increases the neurodegeneration process (and vice versa) in a vicious cycle. (B) Proepileptic/antiepileptic properties of dopamine conditioned by its binding to the D1/D2 family of receptors. Binding of dopamine to D1R promotes an increase in cAMP, which leads to the activation of NMDA-Rs and blockage of GLUT1, thus promoting a massive influx of intracellular Ca2+ and a reduction in glutamate reuptake. This gives rise to an increase in neuroinflammation and neuronal hyperexcitability, which in turn increases the neurodegeneration process (and vice versa) in a vicious cycle. Binding of dopamine to D2R inhibits the production of cAMP, thus promoting the opposite effect of that of D1R activation. NE, norepinephrine; ROS, reactive oxygen species. Full article ">Figure 4
Related molecular pathways between Huntington’s disease and epilepsy. (A) General mechanisms by which mHtt leads to the development of seizures. (B) Neuronal excitability via mitochondrial dysfunction derived from the damage promoted by mHtt. mHtt promotes membrane depolarization, massive influx of intracellular Ca2+, and oxidative stress through the induction of mitochondrial dysfunction and microglia activation and the inhibition of astrocyte GLUT1Rs, BDNF, and GABAergic neurons. All this promotes an increase in neuroinflammation and neuronal hyperexcitability, which in turn increases the neurodegeneration process (and vice versa) in a vicious cycle. Full article ">Figure 5
Seizure activity derived from the main pathological molecular pathways of multiple sclerosis. Autoimmune responses promote demyelination and axonal injury, which in turn trigger the activation of microglia, oligodendrocytes, and macrophages, thus initiating neuroinflammation and neurodegeneration. All this increases neuronal hyperexcitability, which in turn increases the neurodegeneration process (and vice versa) in a vicious cycle. Full article ">

14 pages, 464 KiB

Open AccessArticle

Individualized versus Standardized Risk Assessment in Patients at High Risk for Adverse Drug Reactions (The IDrug Randomized Controlled Trial)–Never Change a Running System?

by Katja S. Just, Catharina Scholl, Miriam Boehme, Kathrin Kastenmüller, Johannes M. Just, Markus Bleckwenn, Stefan Holdenrieder, Florian Meier, Klaus Weckbecker and Julia C. Stingl

Pharmaceuticals 2021, 14(10), 1056; https://doi.org/10.3390/ph14101056 - 18 Oct 2021

Viewed by 2647

Abstract

The aim of this study was to compare effects of an individualized with a standardized risk assessment for adverse drug reactions to improve drug treatment with antithrombotic drugs in older adults. A randomized controlled trial was conducted in general practitioner (GP) offices. Patients [...] Read more.

The aim of this study was to compare effects of an individualized with a standardized risk assessment for adverse drug reactions to improve drug treatment with antithrombotic drugs in older adults. A randomized controlled trial was conducted in general practitioner (GP) offices. Patients aged 60 years and older, multi-morbid, taking antithrombotic drugs and at least one additional drug continuously were randomized to individualized and standardized risk assessment groups. Patients were followed up for nine months. A composite endpoint defined as at least one bleeding, thromboembolic event or death reported via a trigger list was used. Odds ratios (OR) and 95% confidence intervals (CI) were calculated. In total, N = 340 patients were enrolled from 43 GP offices. Patients in the individualized risk assessment group met the composite endpoint more often than in the standardized group (OR 1.63 [95%CI 1.02–2.63]) with multiple adjustments. The OR was higher in patients on phenprocoumon treatment (OR 1.99 [95%CI 1.05–3.76]), and not significant on DOAC treatment (OR 1.52 [95%CI 0.63–3.69]). Pharmacogenenetic variants of CYP2C9, 2C19 and VKORC1 were not observed to be associated with the composite endpoint. The results of this study may indicate that the time point for implementing individualized risk assessments is of importance. Full article

(This article belongs to the Special Issue Clinical Utility of Applying PGx and Deprescribing-Based Decision Support in Polypharmacy: Future Perspectives)

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15 pages, 1252 KiB

Open AccessArticle

In Vitro Effect of Taraxacum officinale Leaf Aqueous Extract on the Interaction between ACE2 Cell Surface Receptor and SARS-CoV-2 Spike Protein D614 and Four Mutants

by Hoai Thi Thu Tran, Michael Gigl, Nguyen Phan Khoi Le, Corinna Dawid and Evelyn Lamy

Pharmaceuticals 2021, 14(10), 1055; https://doi.org/10.3390/ph14101055 - 17 Oct 2021

Cited by 16 | Viewed by 32362

Abstract

To date, there have been rapidly spreading new SARS-CoV-2 “variants of concern”. They all contain multiple mutations in the ACE2 receptor recognition site of the spike protein, compared to the original Wuhan sequence, which is of great concern, because of their potential for [...] Read more.

To date, there have been rapidly spreading new SARS-CoV-2 “variants of concern”. They all contain multiple mutations in the ACE2 receptor recognition site of the spike protein, compared to the original Wuhan sequence, which is of great concern, because of their potential for immune escape. Here we report on the efficacy of common dandelion (Taraxacum officinale) to block protein–protein interaction of SARS-COV-2 spike to the human ACE2 receptor. This could be shown for the wild type and mutant forms (D614G, N501Y, and a mix of K417N, E484K, and N501Y) in human HEK293-hACE2 kidney and A549-hACE2-TMPRSS2 lung cells. High-molecular-weight compounds in the water-based extract account for this effect. Infection of the lung cells using SARS-CoV-2 spike D614 and spike Delta (B.1.617.2) variant pseudotyped lentivirus particles was efficiently prevented by the extract and so was virus-triggered pro-inflammatory interleukin 6 secretion. Modern herbal monographs consider the usage of this medicinal plant as safe. Thus, the in vitro results reported here should encourage further research on the clinical relevance and applicability of the extract as prevention strategy for SARS-CoV-2 infection in terms of a non-invasive, oral post-exposure prophylaxis. Full article

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

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Metabolic analysis of T. officinale (TO) and C. intybus (CI) leaf extract using UPLC-TOF-MS. Measurements were done in high resolution mode with negative electrospray ionization (ESI−) and positive electrospray ionization (ESI+). Full article ">Figure 2
Effect of T. officinale and C. intybus extracts on SARS-CoV-2-Spike–ACE 2 inhibition. (A,B) Concentration-dependent effect of T. officinale (TO) and C. intybus (CI) extract. (C,D). Effect of fractions from TO and CI leaf extract. The extracts were freeze-dried and a molecular weight fractionation was subsequently carried out. The cut-off was set to 5 kDa (HMW > 5 kDa, LMW < 5kDa). H+L: HMW and LMW fractions; 50 mg of dried leaves per mL water was used as reference. HMW and LMW fraction quantities equivalent to dried leaves were used. The binding inhibition was assessed using ELISA technique. Bars are means + SD. Solvent control: distilled water (a.d.); * p < 0.05, ** p < 0.01. Significance of difference was calculated relative to the solvent control by one-way ANOVA. Full article ">Figure 3
Binding inhibition of S1 spike protein to human HEK293-hACE2 cells by extract pre-incubation. Cells were pre-incubated for the indicated times with 10 mg/mL T. officinale (TO), its HMW fraction, equal to 10 mg/mL extract (HMW), and 10 mg/mL C. intybus (CI) or solvent control (a.d.) and subsequently treated with His-tagged S1 spike protein for 1 h without a washing step in between at 4 °C. Binding inhibition was assessed using flow cytometry. N = 3, bars are means + SD. Upper left: cytogram of gated HEK-hACE2 cells. Middle: overlay of representative fluorescence intensity histograms for ACE2 surface expression. Upper right: overlay of representative fluorescence intensity histograms for spike-binding inhibition by the extracts or a.d.; positive control: 20 µg/mL soluble hACE2. Cells were stained with anti-His-tag Alexa Fluor 647 conjugated monoclonal antibody; ** p < 0.01. Significance of difference was calculated relative to the solvent control by one-way ANOVA. Full article ">Figure 4
Binding inhibition of spike D614, and its mutants D614G, N501Y or mix (N501Y, K417N and E484K) to human HEK293-hACE2 and A549-hACE2-TMPRSS2 cells by extract pre- or post-incubation. Overlay of fluorescence intensity histogram for (A) unstained HEK cells, staining control (anti-His-tag Alexa Fluor 647), and cells incubated with His-tag-labelled spike D614, D614G or N501Y for 1 h at 4 °C. (B,C) cells pre-incubated with solvent control (a.d.), 10 mg/mL T. officinale (TO) or 10 mg/mL C. intybus (CI) for 30–60 s, and then treated with His-tag-labelled S1 spike D614, D614G or N501Y protein for 1 h without a washing step in between at 4 °C. (D–G) Effect of extract incubation on HEK or A549 cells either before or after incubation with His-tag-labelled spike D614, D614G, N501Y or mix (N501Y, K417N and E484K) protein at 37 °C. (H) Plant extracts were incubated in saliva from four human donors for 0.5 h at 37 °C. Afterwards, cells were pre-treated with 5 mg/mL extracts for 60 s at 37 °C before incubation with His-tag-labelled spike D614 protein for 0.5 h at 37 °C. Spike-binding inhibition to human cells was assessed using flow cytometric analysis of cells stained with anti-His-tag Alexa Fluor 647 conjugated monoclonal antibody. Bars are means +SD; * p < 0.05, ** p < 0.01. Significance of difference was calculated relative to the respective solvent control by one-way ANOVA. Full article ">Figure 5
Effect of T. officinale extract on ACE2 enzyme activity and protein expression. (A) Viability of A549-hACE2-TMPRSS2 cells was determined using trypan blue cell staining after 84 h exposure to the extract. (B) Cells were incubated with TO extract or 500 ng/mL S1 protein and analyzed for enzyme activity using a fluorescence kit. (C,D) Cells were exposed for 6 h or 24 h to extract without (white bars) or with (black bars) 500 ng/mL S1 protein and analyzed for ACE2 protein expression using a human ACE2 ELISA kit; a.d.: solvent control. Bars are means + SD, N ≥ 3 independent experiments; * p < 0.05, ** p < 0.01. Significance of difference was calculated relative to the respective control by one-way ANOVA. Full article ">Figure 6
Viral transduction inhibition of A549-hACE2-TMPRSS2 cells by T. officinale extract. (A) Cells were pre-treated with T. officinale (TO) or HMW extract for 0.5 h before infection with 7500 TU/mL SARS-CoV-2 spike D614 or Delta (B.1.617.2) variant for 24 h; (B,C) Cells were transduced with 7500 TU/mL SARS-CoV-2 for (B) 3 h before addition of TO for another 21 h or (C) 24 h. After transduction, the medium was exchanged with fresh medium containing TO or HMW extract at the indicated concentrations and post-incubated for 60 h. (D) Cells were pre-treated with 40 mg/mL TO for 3 h before transduction with the indicated virus titer for 24 h. After that, the medium was exchanged with fresh medium and incubated for another 48 h. Luminescence was then detected within 1 h. 0.35 mg/mL HMW extract equals to 20 mg/mL TO extract. Transduction control: (−) negative control: bald lentiviral pseudovirion; (+) positive control: firefly luciferase lentivirus; inhibitor positive control: 100 µg/mL anti-hACE2 antibody. (E) Pro-inflammatory IL-6 cytokine secretion analysis was done either after 24 h virus transduction together with extract (left), after 24 h + 60 h post-infection with extract (middle) or after 60 h post-infection with extract (right) using multiplexing flow cytometric analysis. Solvent control: distilled water (a.d.). N ≥ 3 independent experiments; * p < 0.05, ** p < 0.01. Significance of difference was calculated relative to the solvent control by one-way ANOVA. Full article ">

13 pages, 312 KiB

Open AccessReview

The Impact of SARS-CoV-2 Infection, and Application of Immunosuppressive Agents in Kidney Transplant Recipients Suffering from COVID-19

by Horng-Ta Tseng, Xiang-Chi Wu, Chun-Yao Huang, Chun-Ming Shih, Yi-Wen Lin and Feng-Yen Lin

Pharmaceuticals 2021, 14(10), 1054; https://doi.org/10.3390/ph14101054 - 17 Oct 2021

Cited by 4 | Viewed by 2673

Abstract

In December 2019, the COVID-19 pandemic began to ravage the world quickly, causing unprecedented losses in human life and the economy. A statistical study revealed that the proportion of solid organ transplant (SOT) recipients with severe symptoms and deaths after being infected by [...] Read more.

In December 2019, the COVID-19 pandemic began to ravage the world quickly, causing unprecedented losses in human life and the economy. A statistical study revealed that the proportion of solid organ transplant (SOT) recipients with severe symptoms and deaths after being infected by SARS-CoV-2 is considerably higher than that of non-SOT recipients, and the prognosis is relatively poor. In addition, the clinical manifestation of SOT recipients suffering from COVID-19 is different from that of general COVID-19 patients. Acute kidney injury (AKI) is a common complication in COVID-19 patients, and it is likely more common among SOT recipients infected with SARS-CoV-2. Clinical experts consider that SOT recipients have long-term treatment with immunosuppressants, and the comorbidities are driven by a high rate of severe symptoms and mortality. Orthotopic kidney allograft transplantation is an effective treatment for patients suffering from end-stage kidney disease/kidney failure through which they can easily extend their life. Indeed, kidney transplant recipients have suffered significant damage during this pandemic. To effectively reduce the severity of symptoms and mortality of kidney transplant recipients suffering from COVID-19, precise application of various drugs, particularly immunosuppressants, is necessary. Therefore, herein, we will collate the current clinical experience of treating COVID-19 infection in kidney transplant recipients and discuss the adjustment of patients using immunosuppressive agents in the face of COVID-19. Full article

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

26 pages, 1884 KiB

Open AccessReview

Mucin1 and Mucin16: Therapeutic Targets for Cancer Therapy

by Dong-Hee Lee, Seunghyun Choi, Yoon Park and Hyung-seung Jin

Pharmaceuticals 2021, 14(10), 1053; https://doi.org/10.3390/ph14101053 - 17 Oct 2021

Cited by 48 | Viewed by 10373

Abstract

The mucin (MUC) family is a group of highly glycosylated macromolecules that are abundantly expressed in mammalian epithelial cells. MUC proteins contribute to the formation of the mucus barrier and thus have protective functions against infection. Interestingly, some MUC proteins are aberrantly expressed [...] Read more.

The mucin (MUC) family is a group of highly glycosylated macromolecules that are abundantly expressed in mammalian epithelial cells. MUC proteins contribute to the formation of the mucus barrier and thus have protective functions against infection. Interestingly, some MUC proteins are aberrantly expressed in cancer cells and are involved in cancer development and progression, including cell growth, proliferation, the inhibition of apoptosis, chemoresistance, metabolic reprogramming, and immune evasion. With their unique biological and structural features, MUC proteins have been considered promising therapeutic targets and also biomarkers for human cancer. In this review, we discuss the biological roles of the transmembrane mucins MUC1 and MUC16 in the context of hallmarks of cancer and current efforts to develop MUC1- and MUC16-targeted therapies. Full article

(This article belongs to the Special Issue Anticancer Drugs 2021)

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26 pages, 2880 KiB

Open AccessArticle

Synthesis, Computational Analysis, and Antiproliferative Activity of Novel Benzimidazole Acrylonitriles as Tubulin Polymerization Inhibitors: Part 2

by Anja Beč, Lucija Hok, Leentje Persoons, Els Vanstreels, Dirk Daelemans, Robert Vianello and Marijana Hranjec

Pharmaceuticals 2021, 14(10), 1052; https://doi.org/10.3390/ph14101052 - 17 Oct 2021

Cited by 9 | Viewed by 3311

Abstract

We used classical linear and microwave-assisted synthesis methods to prepare novel N-substituted, benzimidazole-derived acrylonitriles with antiproliferative activity against several cancer cells in vitro. The most potent systems showed pronounced activity against all tested hematological cancer cell lines, with favorable selectivity towards normal [...] Read more.

We used classical linear and microwave-assisted synthesis methods to prepare novel N-substituted, benzimidazole-derived acrylonitriles with antiproliferative activity against several cancer cells in vitro. The most potent systems showed pronounced activity against all tested hematological cancer cell lines, with favorable selectivity towards normal cells. The selection of lead compounds was also tested in vitro for tubulin polymerization inhibition as a possible mechanism of biological action. A combination of docking and molecular dynamics simulations confirmed the suitability of the employed organic skeleton for the design of antitumor drugs and demonstrated that their biological activity relies on binding to the colchicine binding site in tubulin. In addition, it also underlined that higher tubulin affinities are linked with (i) bulkier alkyl and aryl moieties on the benzimidazole nitrogen and (ii) electron-donating substituents on the phenyl group that allow deeper entrance into the hydrophobic pocket within the tubulin’s ?-subunit, consisting of Leu255, Leu248, Met259, Ala354, and Ile378 residues. Full article

(This article belongs to the Section Medicinal Chemistry)

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Graphical abstract

Graphical abstract
Full article ">Figure 1
Benzimidazole-derived tubulin polymerization inhibitors: (a) Nocodazole; (b) 2-aryl-1,2,4-oxadiazolo-benzimidazole derivatives; (c) benzimidazole-2-urea derivatives. Full article ">Figure 2
Benzimidazole acrylonitriles I and II as tubulin polymerization inhibitors. Full article ">Figure 3
Effects of derivatives 50, 64, 68, and 69 on normal PBMC. Apoptosis induction in PBMC from two healthy donors was determined by staining with IncuCyte® Caspase 3/7 Green Reagent and PI followed by live cell monitoring. The percentages of live (white), dead (red), and apoptotic (blue) cells after 72 h are shown (means ± standard error bars). Full article ">Figure 4
Dose-dependent effects of compounds 50, 64, 68, and 69 on in vitro tubulin polymerization. Purified porcine neuronal tubulin and GTP were mixed in a 96-well plate. Docetaxel and combretastatin A4 (CA4) (1 μM) were used as controls for tubulin-stabilizing and tubulin-destabilizing agents, respectively, while DMSO was used as a negative control. The effects on tubulin assembly were monitored in a Tecan Spark multimode plate reader at one minute intervals for one hour at 37 °C. Each condition was tested in duplicate. The level of polymerization was measured by an increase in fluorescence emission intensity at λem = 435 nm. Full article ">Figure 5
Chemical structures of reference systems m1, m2, and colchicine, as discussed in the text. Full article ">Figure 6
Representative structure of the E-isomer of 64 within the colchicine binding site (top) and relative contributions of individual residues to the overall binding free energy (bottom, in %), which lists all residues with favorable contributions higher than –0.5 kcal mol−1 (in blue) and unfavorable contributions exceeding +0.1 kcal mol−1 (in red). Full article ">Scheme 1
Synthesis of benzimidazole acrylonitriles 32–71. Full article ">

13 pages, 2171 KiB

Open AccessArticle

Identification of Novel Anthracycline Resistance Genes and Their Inhibitors

by Onat Kadioglu, Mohamed Elbadawi, Edmond Fleischer and Thomas Efferth

Pharmaceuticals 2021, 14(10), 1051; https://doi.org/10.3390/ph14101051 - 16 Oct 2021

Cited by 4 | Viewed by 3269

Abstract

Differentially expressed genes have been previously identified by us in multidrug-resistant tumor cells mainly resistant to doxorubicin. In the present study, we exemplarily focused on some of these genes to investigate their causative relationship with drug resistance. HMOX1, NEIL2, and PRKCA [...] Read more.

Differentially expressed genes have been previously identified by us in multidrug-resistant tumor cells mainly resistant to doxorubicin. In the present study, we exemplarily focused on some of these genes to investigate their causative relationship with drug resistance. HMOX1, NEIL2, and PRKCA were overexpressed by lentiviral-plasmid-based transfection of HEK293 cells. An in silico drug repurposing approach was applied using virtual screening and molecular docking of FDA-approved drugs to identify inhibitors of these new drug-resistant genes. Overexpression of the selected genes conferred resistance to doxorubicin and daunorubicin but not to vincristine, docetaxel, and cisplatin, indicating the involvement of these genes in resistance to anthracyclines but not to a broader MDR phenotype. Using virtual drug screening and molecular docking analyses, we identified FDA-approved compounds (conivaptan, bexarotene, and desloratadine) that were interacting with HMOX1 and PRKCA at even stronger binding affinities than 1-(adamantan-1-yl)-2-(1H-imidazol-1-yl)ethenone and ellagic acid as known inhibitors of HMOX1 and PRKCA, respectively. Conivaptan treatment increased doxorubicin sensitivity of both HMOX1- and PRKCA-transfected cell lines. Bexarotene treatment had a comparable doxorubicin-sensitizing effect in HMOX1-transfected cells and desloratadine in PRKCA-transfected cells. Novel drug resistance mechanisms independent of ABC transporters have been identified that contribute to anthracycline resistance in MDR cells. Full article

(This article belongs to the Special Issue Identification of Novel Compounds against Multidrug-Resistant Cancer Cells)

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Figure 1
Effect of MDR-related mechanisms on cancer progression. Full article ">Figure 2
Transfection of HEK293 cells with HMOX1, NEIL2, and PRKCA plasmid constructs. (A) Verification of the clones after EcoRI digestion. (B) Verification of transfection with GFP signal observation under a fluorescent microscope. (C) Verification of protein overexpression for HMOX1, NEIL2, and PRKCA. β-actin was used as a loading control. NC, non-transfected control. Full article ">Figure 3
Effect of HMOX1, NEIL2, and PRKCA overexpression on doxorubicin and daunorubicin resistance. Dose response curves for (A) doxorubicin and (B) daunorubicinIC50 values for (C) doxorubicin and (D) daunorubicin treatment. Full article ">Figure 4
Similarity-based analysis of (A) top 10 FDA-approved drugs after HMOX1 screening and (B) top 10 FDA-approved drugs after PRKCA screening. Full article ">Figure 5
Molecular docking poses of conivaptan (blue) and bexarotene (green) on (A) HMOX1 and that of conivaptan (blue) and desloratadine (green) on (B) PRKCA. The known HMOX1 inhibitor 1-(adamantan-1-yl)-2-(1H-imidazol-1-yl)ethenone and the known PRKCA inhibitor ellagic acid are displayed in red. Amino acid residues forming hydrogen bonds are displayed in bold. Full article ">Figure 5 Cont.
Molecular docking poses of conivaptan (blue) and bexarotene (green) on (A) HMOX1 and that of conivaptan (blue) and desloratadine (green) on (B) PRKCA. The known HMOX1 inhibitor 1-(adamantan-1-yl)-2-(1H-imidazol-1-yl)ethenone and the known PRKCA inhibitor ellagic acid are displayed in red. Amino acid residues forming hydrogen bonds are displayed in bold. Full article ">Figure 6
Effect of candidate inhibitors of HMOX1 or PRKCA on doxorubicin cytotoxicity in HMOX1- or PRKCA-transfected cell lines (cell viability, % of control: y-axis; concentration in µM: x-axis). (A) Conivaptan alone, (B) conivaptan with or without doxorubicin in HMOX1-transfected cells, (C) conivaptan with or without doxorubicin in PRKCA-transfected cells, (D) bexarotene alone, (E) bexarotene with or without doxorubicin in HMOX1-transfected cells, (F) desloratadine alone, and (G) desloratadine with or without doxorubicin in PRKCA-transfected cells. Full article ">

20 pages, 2023 KiB

Open AccessReview

Extracellular Vesicles in Human Milk

by Yong Hu, Johannes Thaler and Rienk Nieuwland

Pharmaceuticals 2021, 14(10), 1050; https://doi.org/10.3390/ph14101050 - 15 Oct 2021

Cited by 40 | Viewed by 5725

Abstract

Milk supports the growth and development of infants. An increasing number of mostly recent studies have demonstrated that milk contains a hitherto undescribed component called extracellular vesicles (EVs). This presents questions regarding why milk contains EVs and what their function is. Recently, we [...] Read more.

Milk supports the growth and development of infants. An increasing number of mostly recent studies have demonstrated that milk contains a hitherto undescribed component called extracellular vesicles (EVs). This presents questions regarding why milk contains EVs and what their function is. Recently, we showed that EVs in human milk expose tissue factor, the protein that triggers coagulation or blood clotting, and that milk-derived EVs promote coagulation. Because bovine milk, which also contains EVs, completely lacks this coagulant activity, important differences are present in the biological functions of human milk-derived EVs between species. In this review, we will summarize the current knowledge regarding the presence and biochemical composition of milk EVs, their function(s) and potential clinical applications such as in probiotics, and the unique problems that milk EVs encounter in vivo, including survival of the gastrointestinal conditions encountered in the newborn. The main focus of this review will be human milk-derived EVs, but when available, we will also include information regarding non-human milk for comparison. Full article

(This article belongs to the Special Issue Extracellular Vesicles: Biology and Emerging Therapeutic Opportunities)

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16 pages, 854 KiB

Open AccessEditor’s ChoiceArticle

Side Effects of mRNA-Based COVID-19 Vaccines among Young Adults (18–30 Years Old): An Independent Post-Marketing Study

by Abanoub Riad, Andrea Pokorná, Jitka Klugarová, Natália Antalová, Lucia Kantorová, Michal Koščík and Miloslav Klugar

Pharmaceuticals 2021, 14(10), 1049; https://doi.org/10.3390/ph14101049 - 15 Oct 2021

Cited by 33 | Viewed by 7150

Abstract

Young adults had been widely perceived as a low-risk group for COVID-19 severity; therefore, they were deprioritised within the mass vaccination strategies as their prognosis of COVID-19 infection is relatively more favourable than older age groups. On the other hand, vaccination of this [...] Read more.

Young adults had been widely perceived as a low-risk group for COVID-19 severity; therefore, they were deprioritised within the mass vaccination strategies as their prognosis of COVID-19 infection is relatively more favourable than older age groups. On the other hand, vaccination of this demographic group is indispensable to achieve herd immunity. A cross-sectional survey-based study was used to evaluate the side effects of mRNA-based COVID-19 vaccines among university students in the Czech Republic. The validated questionnaire was delivered in a digital form, and it consisted of demographic data; COVID-19 vaccine-related anamnesis; and local, systemic, orofacial, and skin-related side effects’ prevalence, onset, and duration. Out of the 539 included participants, 70.1% were females and 45.8% were <23 years old. The vast majority (95.2%) reported at least one side effect. The most common side effect was injection site pain (91.8%), followed by fatigue (62.5%), headache (36.4%), and muscle pain (34.9%). The majority of local side effects occurred after both doses (74.4%), while most systemic side effects occurred after the second dose only (56.2%). Most local (94.2%) and systemic (93.3%) side effects resolved within three days after vaccination. Females participants’ adjusted odds ratio (AOR) showed they were 2.566 (CI 95%: 1.103–5.970) times more likely to experience post-vaccination side effects, and the participants who received two doses reported an increased AOR of 1.896 (0.708–5.077) for experiencing side effects. The results of this study imply that mRNA-based COVID-19 vaccines are highly probably safe for young adults, and further studies are required to investigate the role of medical anamnesis, prior COVID-19 infection, and gender in side effects incidence. Full article

(This article belongs to the Section Biopharmaceuticals)

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19 pages, 4584 KiB

Open AccessArticle

SARS-CoV-2 Fears Green: The Chlorophyll Catabolite Pheophorbide A Is a Potent Antiviral

by Guillermo H. Jimenez-Aleman, Victoria Castro, Addis Londaitsbehere, Marta Gutierrez-Rodríguez, Urtzi Garaigorta, Roberto Solano and Pablo Gastaminza

Pharmaceuticals 2021, 14(10), 1048; https://doi.org/10.3390/ph14101048 - 15 Oct 2021

Cited by 18 | Viewed by 7090

Abstract

SARS-CoV-2 pandemic is having devastating consequences worldwide. Although vaccination advances at good pace, effectiveness against emerging variants is unpredictable. The virus has displayed a remarkable resistance to treatments and no drugs have been proved fully effective against COVID-19. Thus, despite the international efforts, [...] Read more.

SARS-CoV-2 pandemic is having devastating consequences worldwide. Although vaccination advances at good pace, effectiveness against emerging variants is unpredictable. The virus has displayed a remarkable resistance to treatments and no drugs have been proved fully effective against COVID-19. Thus, despite the international efforts, there is still an urgent need for new potent and safe antivirals against SARS-CoV-2. Here, we exploited the enormous potential of plant metabolism using the bryophyte Marchantia polymorpha L. and identified a potent SARS-CoV-2 antiviral, following a bioactivity-guided fractionation and mass-spectrometry approach. We found that the chlorophyll derivative Pheophorbide a (PheoA), a porphyrin compound similar to animal Protoporphyrin IX, has an extraordinary antiviral activity against SARS-CoV-2, preventing infection of cultured monkey and human cells, without noticeable cytotoxicity. We also show that PheoA targets the viral particle, interfering with its infectivity in a dose- and time-dependent manner. Besides SARS-CoV-2, PheoA also displayed a broad-spectrum antiviral activity against enveloped RNA viral pathogens such as HCV, West Nile, and other coronaviruses. Our results indicate that PheoA displays a remarkable potency and a satisfactory therapeutic index, which together with its previous use in photoactivable cancer therapy in humans, suggest that it may be considered as a potential candidate for antiviral therapy against SARS-CoV-2. Full article

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

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Graphical abstract

Graphical abstract
Full article ">Figure 1
M. polymorpha extracts interfere with SARS-CoV-2-induced cytopathic effect and virus propagation. (A,B) Vero E6 cells were inoculated with SARS-CoV-2 (MOI = 0.001) in the presence of serial 2-fold dilutions of crude extracts from (A) two different M. polymorpha ecotypes, ruderalis (Ex1) and polymorpha (Ex2) or (B) WT, Mpcoi1-2 or Mpc1hdz Marchantia plants, incubated for 72 h, time after which they were fixed and stained with a crystal violet solution. Mock-infected cells were used as the control of the integrity of the cell monolayer. Images show a representative experimental plate comparing M. polymorpha extracts with vehicle (DMSO)-treated cells. (C) Vero E6 cells were inoculated with SARS-CoV-2 (MOI = 0.001) in the presence of vehicle (DMSO), RMDV (25 µM) or a 100 µg/mL dilution of crude extracts. Uninfected samples were used as the control (mock). Total RNA was extracted 72 h post-infection and subjected to RT-qPCR to determine viral load. The dotted line indicates the limit of detection of the assay. Normalized viral RNA levels are shown as percentage of the viral RNA found in vehicle-treated cells. Data are shown as mean (± SD) of three biological replicates. Statistical significance was estimated using one-way ANOVA and a Dunnet’s post hoc test (* p < 0.05). Full article ">Figure 2
Extract fractionation and identification of antiviral fraction pools. (A) Vero E6 cells were inoculated with SARS-CoV-2 (MOI = 0.001) in the presence of serial 2-fold dilutions of vehicle (DMSO), a crude Marchantia extract and the fraction pools. Inoculated cultures were incubated for 72 h, time after which they were fixed and stained with a crystal violet solution. Mock-treated cells were used as the control of the integrity of the cell monolayer (non-infected). (B) Vero E6 cells were inoculated (MOI = 0.01) in the presence of serial dilution of the fractions and incubated for 24 h before fixation; processing for immunofluorescence microscopy and cytotoxicity assays was as described in the materials and methods section. Representative images of the fraction pool cell-based evaluation are shown. (C) Quantitation of infection efficiency, cell number and cell viability. These data are shown as average (± SD) of three biological replicates. Statistical significance was estimated using one-way ANOVA (Dunnet´s post hoc test, * p < 0.05). Full article ">Figure 3
Identification of the main antiviral compound in Marchantia extracts. (A) Representative TLC of Marchantia WT and c1hdz extracts. Compounds C, D and E (box) were tested for their antiviral potential. (B) Vero E6 cells were inoculated (MOI = 0.01) in the presence of the indicated compounds and incubated for 24 h before fixation and processing for immunofluorescence microscopy. Data are shown as average (± SD) of three biological replicates. Statistical significance was estimated using one-way ANOVA and a Dunnet´s post hoc test (* p < 0.05). (C) Representative HPLC/MS analysis (shown for WT) of fractions C, D, and E, including exact mass determination of the antiviral candidate 1, and its inferred chemical structure. Full article ">Figure 4
PheoA shows antiviral activity against SARS-CoV-2 in Vero E6 cells and human lung epithelial A549-ACE2 and Calu3 cell lines. (A–C) Commercially available PheoA was serially diluted and mixed (1:1, v/v) with SARS-CoV-2 preparations to achieve the indicated compound concentrations and a final MOI of 0.005 for (A) Vero E6 and (B) Calu3 and 0.01 for (C) A549-ACE2 cells. Cultures were incubated for 48 h, fixed and processed for automated immunofluorescence microscopy analysis. Parallel, uninfected cultures were processed for cytotoxicity evaluation using an MTT assay. Relative infection efficiency data (n = 3 per dose) are shown as individual data and a PROBIT regression curve (green line) using the represented values. Cytotoxicity data (n = 3 per dose) are shown as the individual data and a moving average trend line. (D) A549-ACE2 cells were inoculated at MOI = 0.01 in the presence of increasing concentrations of PheoA or RMDV (5 µM) and incubated for 48 h. Samples of the supernatants were collected, heat-inactivated and directly subjected to RT-qPCR to estimate overall infection efficiency. Data are expressed as relative values compared with the vehicle (DMSO)-treated cells and are shown as the mean (± SD) of three biological replicates (n = 3). Statistical significance was estimated using one-way ANOVA and a Dunnet´s post hoc test (* p < 0.05). Full article ">Figure 5
Antiviral spectrum of PheoA against different RNA viruses. The effectiveness of PheoA was tested against four different recombinant RNA viruses expressing reporter genes. (A) Huh7 cells were infected with HCVtcp in the presence of increasing PheoA doses and luciferase activity was determined 48 h post-inoculation. (B–D) Cells were inoculated in the presence of increasing concentrations of PheoA at MOI 0.01 and incubated to enable virus propagation. At the endpoint, cells were fixed and counter stained with DAPI to control for unexpected cytotoxic effects. Relative infection efficiency was estimated using automated microscopy and is expressed as percentage of the infection efficiency observed in control wells. (B) Huh7 cells were infected with hCoV-229E-GFP and fixed 48 h post-inoculation. (C) Huh7 cells were infected with WNV-GFP and fixed 48 h post-inoculation. (D) A549-ACE2 cells were inoculated with VSV-GFP and fixed 16 h post-inoculation. Individual replicate data are shown as green dots (n = 3) and the PROBIT regression curve used to estimate EC50 values is shown. Full article ">Figure 6
Combination treatment of PheoA with RMDV. Vero E6 cells were inoculated at MOI = 0.005 in the presence of increasing concentrations of PheoA in combination with increasing doses of RMDV. Twenty-four hours post infection, cells were fixed and processed for automated immunofluorescence microscopy. Relative infection efficiency values were estimated as percentage of the values obtained in mock-treated cells. (A) Data are shown as average of two biological replicates. (B) Heatmap describing the areas of synergy within the combination treatments. Full article ">Figure 7
Time-of-addition experiments indicate that PheoA interferes with early aspects of SARS-CoV-2 infection. Vero E6 cells were inoculated at MOI = 5 in the presence (gray) or absence (white) of the indicated doses of PheoA, RMDV or imatinib as described in both the text and the scheme. Cells were incubated for 6 h in the presence (gray) or absence (white) before chemical fixation and processing for immunofluorescence microscopy. (A) Schematic diagram of the times where compound was present in the assay. (B) Infection efficiency is expressed as the percentage of that observed in vehicle DMSO-treated cells and is shown as average and standard deviation of three biological replicates (n = 3). Statistical significance was estimated using one-way ANOVA and a Dunnet´s post hoc test (* p < 0.05). Full article ">Figure 8
PheoA interferes with SARS-CoV-2 virion infectivity in a dose- and time-dependent manner. SARS-CoV-2 virus stocks were diluted to obtain 1 × 105 TCID50/mL and were mixed with increasing concentrations of PheoA or the vehicle (DMSO). (A) Dose-dependent reduction in SARS-CoV-2 infectivity by PheoA. Virus-compound mixtures were incubated at room temperature for 30 min and were serially diluted to determine the remaining infectivity titer using endpoint dilution and determination of virus-induced cytopathic effect by crystal violet staining in Vero-E6 cells. (B) Pre-incubation time-dependent reduction in SARS-CoV-2 infectivity by PheoA. Experiments were carried out using a fixed dose of PheoA (340 nM) and increasing pre-incubation times before serial dilution for TCID50 determination. Values are expressed as LOG TCID50/mL and shown as the average and standard deviation of three independent experiments (n = 3). Statistical significance was estimated using one-way ANOVA and a Dunnet’s post hoc test (* p < 0.05). Full article ">

20 pages, 11500 KiB

Open AccessArticle

Formulation Study of a Co-Processed, Rice Starch-Based, All-in-One Excipient for Direct Compression Using the SeDeM-ODT Expert System

by Karnkamol Trisopon, Nisit Kittipongpatana, Phanphen Wattanaarsakit and Ornanong Suwannapakul Kittipongpatana

Pharmaceuticals 2021, 14(10), 1047; https://doi.org/10.3390/ph14101047 - 14 Oct 2021

Cited by 7 | Viewed by 5110

Abstract

A co-processed, rice starch-based excipient (CS), previously developed and shown to exhibit good pharmaceutical properties, is investigated as an all-in-one excipient for direct compression (DC). An SeDeM-ODT expert system is applied to evaluate the formulation containing CS, in comparison with those containing the [...] Read more.

A co-processed, rice starch-based excipient (CS), previously developed and shown to exhibit good pharmaceutical properties, is investigated as an all-in-one excipient for direct compression (DC). An SeDeM-ODT expert system is applied to evaluate the formulation containing CS, in comparison with those containing the physical mixture and the commercial DC excipients. The results revealed that CS showed acceptable values in all six incidence factors of the SeDeM-ODT diagram. In addition, the comprehensive indices (IGC and IGCB) were higher than 5.0, which indicated that CS could be compressed with DC technique without additional blending with a disintegrant in tablet formulation. The formulation study suggested that CS can be diluted up to 60% in the formulation to compensate for unsatisfactory properties of paracetamol. At this percentage, CS-containing tablets exhibited narrow weight variation (1.5%), low friability (0.43%), acceptable drug content (98%), and rapid disintegration (10 s). The dissolution profile of CS displayed that more than 80% of the drug content was released within 2 min. The functionality of CS was comparable to that of high functionality excipient composite (HFEC), whereas other excipients were unsuccessful in formulating the tablets. These results indicated that CS was a suitable all-in-one excipient for application in DC of tablets. Full article

(This article belongs to the Special Issue Formulation and Evaluation of Tablets of Different Drugs)

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12 pages, 2993 KiB

Open AccessEditor’s ChoiceArticle

Corylin Ameliorates LPS-Induced Acute Lung Injury via Suppressing the MAPKs and IL-6/STAT3 Signaling Pathways

by I-Chen Chen, Shu-Chi Wang, Yi-Ting Chen, Hsin-Han Tseng, Po-Len Liu, Tzu-Chieh Lin, Hsin-En Wu, Yuan-Ru Chen, Yu-Hsin Tseng, Jong-Hau Hsu, Zen-Kong Dai, Jau-Ling Suen and Chia-Yang Li

Pharmaceuticals 2021, 14(10), 1046; https://doi.org/10.3390/ph14101046 - 14 Oct 2021

Cited by 22 | Viewed by 4378

Abstract

Acute lung injury (ALI) is a high mortality disease with acute inflammation. Corylin is a compound isolated from the whole plant of Psoralea corylifolia L. and has been reported to have anti-inflammatory activities. Herein, we investigated the therapeutic potential of corylin on lipopolysaccharides [...] Read more.

Acute lung injury (ALI) is a high mortality disease with acute inflammation. Corylin is a compound isolated from the whole plant of Psoralea corylifolia L. and has been reported to have anti-inflammatory activities. Herein, we investigated the therapeutic potential of corylin on lipopolysaccharides (LPS)-induced ALI, both in vitro and in vivo. The levels of proinflammatory cytokine secretions were analyzed by ELISA; the expressions of inflammation-associated proteins were detected using Western blot; and the number of immune cell infiltrations in the bronchial alveolar lavage fluid (BALF) were detected by multicolor flow cytometry and lung tissues by hematoxylin and eosin (HE) staining, respectively. Experimental results indicated that corylin attenuated LPS-induced IL-6 production in human bronchial epithelial cells (HBEC3-KT cells). In intratracheal LPS-induced ALI mice, corylin attenuated tissue damage, suppressed inflammatory cell infiltration, and decreased IL-6 and TNF-? secretions in the BALF and serum. Moreover, it further inhibited the phosphorylation of mitogen-activated protein kinases (MAPKs), including p-JNK, p-ERK, p-p38, and repressed the activation of signal transducer and activator of transcription 3 (STAT3) in lungs. Collectively, our results are the first to demonstrate the anti-inflammatory effects of corylin on LPS-induced ALI and suggest corylin has significant potential as a novel therapeutic agent for ALI. Full article

(This article belongs to the Special Issue Lung Injury and Repair)

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The effects of corylin on the cell viability and the production of IL-6 by LPS-induced HBEC3-KT cells. Cells were pre-treated with different concentrations of corylin (0~50 μM) for 1 h following treatment of LPS (1 μg/mL) for 24 h. (A) Cell viability was examined by MTT assay. (B) The concentration of IL-6 in the cell culture supernatant was detected using ELISA. The data are presented as means ± SEM of three independent experiments. Statistical significances are represented as follows: *** p < 0.001 and **** p < 0.0001 vs. LPS alone. Full article ">Figure 2
Effects of corylin on LPS-induced inflammatory cytokine productions in BALF. Mice were intraperitoneally injected with corylin (10 mg/kg or 20 mg/kg) for 1 h following intratracheal administration of LPS for 4 h. The mice were sacrificed and the BALF was collected. The expression levels of (A) TNF-α, (B) IL-6, (C) IL-1β, and (D) IL-12 in BALF were analyzed using ELISA. The data are presented as means ± SEM of three independent experiments. Statistical significances are represented as follows: * p < 0.05, and *** p < 0.001 and **** p < 0.0001 vs. LPS alone. Full article ">Figure 3
Effects of corylin treatment on immune cell infiltration in BALFs in LPS-induced mice. BALFs were collected from LPS and/or corylin-treated mice. (A) The total cell numbers in BALFs. The cell subsets were identified by multi-color flow cytometry, including (B) macrophages, (C) granulocytes, and (D) lymphocytes. Results are shown as mean ± SEM. * p < 0.05, ** p < 0.01, and **** p < 0.0001 vs. LPS alone by one-way ANOVA followed by post hoc Tukey’s test. The numbers of mice are pooled from two independent experiments (n = 8~10). Full article ">Figure 4
Effects of corylin on the expression and phosphorylation of MAPKs and STAT3 in lung tissues. Expressions of phospho-JNK, JNK, phospho-ERK, ERK, phospho-p38 MAPK, p38 MAPK, phospho-STAT3 and STAT3 were analyzed by Western blot. (A) The representative blot of triplicate experiments. Quantitated results of (B) phospho-JNK/JNK ratio (n = 6), (C) phospho-ERK/ERK ratio (n = 6), (D) phospho-p38/p38 ratio (n = 6), and (E) phospho-STAT3/STAT3 ratio (n = 9~10) were shown as mean ± SEM. The relative fold of phosphorylation activity was normalized to untreated samples. (* p < 0.05, ** p < 0.01 and *** p < 0.001 vs. LPS alone). Full article ">Figure 5
Effects of corylin on the expression of IL-6 and TNF-α in serum of LPS-induced mice. The expression levels of (A) IL-6 and (B) TNF-α in serum were measured using ELISA. The data are shown as mean ± SEM (n = 7~9). (* p < 0.05 and *** p < 0.001 vs. LPS alone). Full article ">Figure 6
Effects of corylin on the lung in LPS-challenged mice. (A) Representative photographs of the lung tissues stained with HE. Upper left, control, the representative image PBS control. Upper right, the representative image of LPS administration. Lower left, the representative image of low dose corylin (10 mg/kg) pre-treatment following LPS administration. Lower right, the representative image of high dose corylin (20 mg/kg) pre-treatment following LPS administration. (B) Morphological changes in lung sections were semi-quantified using lung injury score. The results showed a significant reduction in the severity of lung injury in mice treatment with corylin compared to the LPS-induced ALI mice (n = 4~6). The magnification is 400X. The data are shown as mean ± SEM. (*** p < 0.001 and **** p < 0.0001 vs. LPS alone). Full article ">Figure 7
Anti-inflammatory effect of corylin on LPS-induced ALI. The experimental results demonstrated that corylin attenuated the overproduction of IL-6 in LPS-activated human bronchial epithelial cells. In intratracheal LPS-induced ALI mice, corylin attenuated tissue damages, suppressed inflammatory cell infiltration, and decreased secretion of IL-6 and TNF-α in the BALF and serum; moreover, it further inhibited the expression of phosphorylation of mitogen-activated protein kinases (MAPKs), including the expression of p-JNK/JNK, p-ERK/ERK, p-p38/p38, and repressed the activation of signal transducer and activator of transcription 3 (STAT3) in lung. Taken together, our results are the first to demonstrate the anti-inflammatory effects of corylin on LPS-induced ALI and suggest corylin has significant potential as a novel therapeutic agent for ALI. Full article ">

18 pages, 9238 KiB

Open AccessArticle

Pioglitazone Is a Mild Carrier-Dependent Uncoupler of Oxidative Phosphorylation and a Modulator of Mitochondrial Permeability Transition

by Ekaterina S. Kharechkina, Anna B. Nikiforova, Konstantin N. Belosludtsev, Tatyana I. Rokitskaya, Yuri N. Antonenko and Alexey G. Kruglov

Pharmaceuticals 2021, 14(10), 1045; https://doi.org/10.3390/ph14101045 - 14 Oct 2021

Cited by 8 | Viewed by 3562

Abstract

Pioglitazone (PIO) is an insulin-sensitizing antidiabetic drug, which normalizes glucose and lipid metabolism but may provoke heart and liver failure and chronic kidney diseases. Both therapeutic and adverse effects of PIO can be accomplished through mitochondrial targets. Here, we explored the capability of [...] Read more.

Pioglitazone (PIO) is an insulin-sensitizing antidiabetic drug, which normalizes glucose and lipid metabolism but may provoke heart and liver failure and chronic kidney diseases. Both therapeutic and adverse effects of PIO can be accomplished through mitochondrial targets. Here, we explored the capability of PIO to modulate the mitochondrial membrane potential (??_m) and the permeability transition pore (mPTP) opening in different models in vitro. ??_m was measured using tetraphenylphosphonium and the fluorescent dye rhodamine 123. The coupling of oxidative phosphorylation was estimated polarographically. The transport of ions and solutes across membranes was registered by potentiometric and spectral techniques. We found that PIO decreased ??_m in isolated mitochondria and intact thymocytes and the efficiency of ADP phosphorylation, particularly after the addition of Ca²⁺. The presence of the cytosolic fraction mitigated mitochondrial depolarization but made it sustained. Carboxyatractyloside diminished the PIO-dependent depolarization. PIO activated proton transport in deenergized mitochondria but not in artificial phospholipid vesicles. PIO had no effect on K⁺ and Ca²⁺ inward transport but drastically decreased the mitochondrial Ca²⁺-retention capacity and protective effects of adenine nucleotides against mPTP opening. Thus, PIO is a mild, partly ATP/ADP-translocase-dependent, uncoupler and a modulator of ATP production and mPTP sensitivity to Ca²⁺ and adenine nucleotides. These properties contribute to both therapeutic and adverse effects of PIO. Full article

(This article belongs to the Special Issue Heterocyclic Compounds and Their Application in Therapy)

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Full article ">Figure 1
Effect of PIO on ΔΨm and the coupling of mitochondria. (A,B) Assessment of the effect of PIO on ΔΨm using TPP+ (A) and rhodamine 123 (Rhod 123, (B)). (C,D) Effect of PIO on ΔΨm in mitochondria in the resting state (C) and during the ADP phosphorylation (D). (A) Rat liver mitochondria (RLM) (1 mg prot./mL) were placed in a standard incubation medium supplemented with 5 mM K+-succinate, 1 mM EGTA, rotenone (2 µg/mL), and 1 µM TPP+. Arrows show the addition of 25 or 100 µM PIO and 500 nM FCCP. (B–D) A mitochondrial suspension (0.75 mg/mL) supplemented with respiratory substrates, 1 mM EGTA, and 330 nM rhodamine 123 was placed in wells with 1% dimethyl sulfoxide (DMSO) (vehicle control) or PIO at indicated concentrations (µM) (B–D), 500 nM FCCP, antimycin A (2.5 µg/mL), and valinomycin (25 ng/mL) (B), and 2 mM ADP (D) just before measurements. Everywhere, except the curve designated “M+P” (5 mM malate + 5 mM pyruvate), the respiratory substrate was 5 mM K+-succinate with the addition of rotenone (2 µg/mL). Numbers on curves are the means ± S.E.M. (n = 3) of one representative experiment of three identical experiments. (B) Where indicated, 500 nM FCCP was added to wells with 50, 100, and 200 µM PIO to calibrate the signal. Insert in panel (C) shows a shift in ΔΨm caused by FCCP, DNP, and PIO after 5 min of incubation. Insert in panel (D) shows the effect of PIO on the respiration rate in state 2 and state 3 and the respiratory control (RC) coefficient. In inserts, numbers on curves are the means ± S.E.M. of three independent experiments (n = 9 (C) and 3 (D)). Full article ">Figure 2
PIO increases mitochondrial sensitivity to Ca2+ and decreases the protective effect of mPTP inhibitors, ATP and ADP. (A–C) Ca2+-dependent mitochondrial swelling in the presence of PIO and ATP/ADP. (D) Effect of PIO on the Ca2+-retention capacity of mitochondria. (A–C) Mitochondrial suspension (0.75 mg prot./mL) supplemented with 5 mM K+-succinate (plus 2 µg/mL rotenone) and, where shown, 2 mM ADP (B) and 2 mM ATP (C) (pH 7.4) were placed in wells with indicated additions: 1 mM EGTA, 1% DMSO, 10–100 µM PIO, and 10 µg/mL oligomycin (Oligo). The arrow shows the addition of 50 µM Ca2+. Representative data of three similar experiments are shown. Points on traces are the means ± S.E.M. (n = 3). (D) Ca2+-retention capacity equal to 100% corresponds to 210.9 ± 23.45 nmol Ca2+/mg protein. Values in columns are means ± S.E.M. (n = 3) of three independent experiments. The asterisk shows the significant difference from the vehicle control (p < 0.05). The insert shows the kinetics of accumulation of 30 µM Ca2+ in the absence (Control) and in the presence of 100 µM PIO. Full article ">Figure 3
Effect of PIO on the capability of mitochondria to support ATP production in the presence of Ca2+ (A) and on the level of matrix ATP (B). Mitochondria (0.2 (A) and 0.5 mg prot./mL (B)) were placed in the standard incubation medium, which contained a 20% solution of a luciferin-luciferase reagent, 5 mM K+-succinate, 2 µg/mL of rotenone, 10 µM EGTA, and, where indicated, 5–50 µM PIO, 1 µM CATR, and 10 µg/mL oligomycin (Oligo). Vertical dashed lines (A) or arrows (B) show the addition of 500 µM ADP, 50 µM Ca2+, and 5 µM ATP standard (to calibrate the chemiluminescent signal). Representative data of three separate experiments are shown. Traces are the means for three wells. Full article ">Figure 4
Mechanism of transmembrane proton transport by PIO. (A) Possible mechanism of proton transport by PIO across the IMM. (B) Comparison of CCCP- and PIO-mediated proton fluxes through liposomes loaded with the pH probe pyranine. The inner liposomal pH value was estimated from the pyranine fluorescence intensity measured at 505 nm upon excitation at 455 nm. Lasalocid A (1 µM) was added approximately at 800 s to equilibrate pH. PIO concentrations were 10 µM, red curve; 50 µM, green curve; 75 µM, blue curve. CCCP concentration was 1 µM (pink curve). Lipid concentration was 20 μg/mL, T = 15 °C. Other conditions: see Materials and Methods Section. The proton flux was initiated by an alkaline pH shift from рН 6 to рН 8, which was caused by the addition of the previously determined aliquot of KOH. In the presence of the protonophores, the pyranine fluorescence gradually increased, indicating the alignment of the pH values inside and outside liposomes due to proton transfer mediated by a protonophore. (C,D) Effect of PIO, palmitic acid, and FCCP on the swelling of deenergized mitochondria in NH4NO3-based medium. Mitochondria (0.75 mg prot./mL) were added to NH4NO3-based medium supplemented with rotenone (2.5 mg/mL), and 1 min later, the suspension was placed in the wells of a plate that contained PIO, sodium palmitate (PA), and FCCP at indicated concentrations. (C) Standard curves of mitochondrial swelling. (D) Initial rates of mitochondrial swelling during the first 3 min of incubation. Points are the means ± S.E.M. of three independent experiments (n = 9). Full article ">Figure 5
Effect of inhibitors of ANT and UCPs on the shift in ΔΨm caused by PIO and DNP. Incubation medium contained indicated concentrations of PIO and DNP, and, where shown, 1 mM GDP and 2 µM CATR. (A,B) Dynamics of ΔΨm changes (decrease) in relation to control (1% DMSO). The data of one representative experiment of three similar experiments are shown. Values on traces are the means for three wells. (C) Effect of GDP and CATR on the average PIO-dependent shift in ΔΨm. Each point on the curves is an average ΔΨm shift defined as a mean ± S.E.M. for 60 points of experimental curves (such as in Panel (A)) for three individual experiments (n = 180). (D) Changes in the DNP-dependent ΔΨm shift caused by CATR and GDP in relation to DNP alone (Control). Full article ">Figure 6
PIO decreases ΔΨm in isolated intact thymocytes. (A–D) Typical plots obtained in the absence of additions (A, Control), in the presence of 50 (B) and 200 µM PIO (C), and 500 nM FCCP (D). (E) The portion of depolarized cells in different experimental groups (means ± SEM, n = 3). * Statistically significant (p < 0.05) differences from control (without PIO). Full article ">Figure 7
Effect of a cytosolic fraction (RLH) on PIO-dependent changes in ΔΨm. Just before measurements, mitochondria alone (RLM) (0.75 mg prot./mL) (A,D) or in combination with 5 (B,D) and 15 mg prot./mL (C,D) of RLH were added to a standard incubation medium supplemented with 5 mM K+-succinate, 1 mM EGTA, rotenone (2.5 µg/mL), and 330 nM rhodamine 123, and then transferred to the wells of a 96-well plate, which contained 1% DMSO (Control), 10–200 µM PIO, or 500 nM FCCP, antimycin A (2.5 µg/mL), and valinomycin (25 ng/mL). Traces are the differences in ΔΨm between PIO- and DMSO-containing samples in one representative experiment of three similar experiments. Full article ">Figure 8
Contribution of uncoupling and mPTP-modulating effects of PIO to its healing and harmful action in the cell and the organism. CVD: cardiovascular disease. Full article ">

16 pages, 2600 KiB

Open AccessFeature PaperArticle

Esculetin Provides Neuroprotection against Mutant Huntingtin-Induced Toxicity in Huntington’s Disease Models

by Letizia Pruccoli, Carlo Breda, Gabriella Teti, Mirella Falconi, Flaviano Giorgini and Andrea Tarozzi

Pharmaceuticals 2021, 14(10), 1044; https://doi.org/10.3390/ph14101044 - 13 Oct 2021

Cited by 7 | Viewed by 2968

Abstract

Huntington’s disease (HD) is a neurodegenerative disorder caused by an abnormal CAG trinucleotide repeat expansion within exon 1 of the huntingtin (HTT) gene. This mutation leads to the production of mutant HTT (mHTT) protein which triggers neuronal death through several mechanisms. Here, we [...] Read more.

Huntington’s disease (HD) is a neurodegenerative disorder caused by an abnormal CAG trinucleotide repeat expansion within exon 1 of the huntingtin (HTT) gene. This mutation leads to the production of mutant HTT (mHTT) protein which triggers neuronal death through several mechanisms. Here, we investigated the neuroprotective effects of esculetin (ESC), a bioactive phenolic compound, in an inducible PC12 model and a transgenic Drosophila melanogaster model of HD, both of which express mHTT fragments. ESC partially inhibited the progression of mHTT aggregation and reduced neuronal death through its ability to counteract the oxidative stress and mitochondria impairment elicited by mHTT in the PC12 model. The ability of ESC to counteract neuronal death was also confirmed in the transgenic Drosophila model. Although ESC did not modify the lifespan of the transgenic Drosophila, it still seemed to have a positive impact on the HD phenotype of this model. Based on our findings, ESC may be further studied as a potential neuroprotective agent in a rodent transgenic model of HD. Full article

(This article belongs to the Special Issue Natural Bioactive Compounds and Neurological Disorders: Exploiting Resources from Nature)

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25 pages, 6443 KiB

Open AccessReview

Recent Developments in the Synthesis of ?-Diketones

by Gonzalo de Gonzalo and Andrés R. Alcántara

Pharmaceuticals 2021, 14(10), 1043; https://doi.org/10.3390/ph14101043 - 13 Oct 2021

Cited by 29 | Viewed by 7901

Abstract

Apart from being one of the most important intermediates in chemical synthesis, broadly used in the formation of C–C bonds among other processes, the ?-dicarbonyl structure is present in a huge number of biologically and pharmaceutically active compounds. In fact, mainly derived from [...] Read more.

Apart from being one of the most important intermediates in chemical synthesis, broadly used in the formation of C–C bonds among other processes, the ?-dicarbonyl structure is present in a huge number of biologically and pharmaceutically active compounds. In fact, mainly derived from the well-known antioxidant capability associated with the corresponding enol tautomer, ?-diketones are valuable compounds in the treatment of many pathological disorders, such as cardiovascular and liver diseases, hypertension, obesity, diabetes, neurological disorders, inflammation, skin diseases, fibrosis, or arthritis; therefore, the synthesis of these structures is an area of overwhelming interest for organic chemists. This paper is devoted to the advances achieved in the last ten years for the preparation of 1,3-diketones, using different chemical (Claisen, hydration of alkynones, decarboxylative coupling) or catalytic (biocatalysis, organocatalytic, metal-based catalysis) methodologies: Additionally, the preparation of branched ?-dicarbonyl compounds by means of ?-functionalization of non-substituted 1,3-diketones are also discussed. Full article

(This article belongs to the Special Issue β-Diketones and Their Derivatives: Synthesis, Characterization and Biomedical Applications)

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10 pages, 1799 KiB

Open AccessArticle

Ten-Year Outcomes of Intravitreal Bevacizumab for Myopic Choroidal Neovascularization: Analysis of Prognostic Factors

by Fabiana Mallone, Rosalia Giustolisi, Federica Franzone, Marco Marenco, Rocco Plateroti, Marcella Nebbioso, Alessandro Lambiase and Magda Gharbiya

Pharmaceuticals 2021, 14(10), 1042; https://doi.org/10.3390/ph14101042 - 13 Oct 2021

Cited by 6 | Viewed by 3602

Abstract

The current standard treatment of myopic choroidal neovascularisation (mCNV) is intravitreal injection of VEGF antagonists. This study was proposed to assess efficacy and safety of intravitreal bevacizumab (IVB) for the treatment of mCNV across a 10-year follow-up. Thirty eyes of thirty patients with [...] Read more.

The current standard treatment of myopic choroidal neovascularisation (mCNV) is intravitreal injection of VEGF antagonists. This study was proposed to assess efficacy and safety of intravitreal bevacizumab (IVB) for the treatment of mCNV across a 10-year follow-up. Thirty eyes of thirty patients with treatment-naïve mCNV who underwent IVB and were followed up with for a minimum of ten years were recruited for the present retrospective cohort study. All participants were treated with three monthly IVB at baseline and then evaluated and treated under pro re nata (PRN) schedule. Outcome measures were to determine BCVA changes over years and identify the predictive factors of both final visual outcome and need for retreatment. Analysis of the main involved prognostic factors with correlations among variables is reported. Visual acuity remained stable at 10-year follow-up (p = 0.001) with the greatest improvement at 2 years (p < 0.0001) in all CNV locations. Baseline BCVA correlated positively with final BCVA (? = 0.88, p < 0.0001, R²: 0.75). No predictive factors for the need of additional injections were identified. Retinal and choroidal thickness significantly reduced over time but without correlation with the number of injections. CNV max height and area significantly decreased at 10 years (p < 0.0001 and p = 0.003, respectively), with complete regression of mCNV lesion in 40% of subjects. Intravitreal bevacizumab resulted as long-term effective and safe therapy for mCNV with sustained results at 10 years. Full article

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

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19 pages, 769 KiB

Open AccessReview

Plasma Rich in Growth Factors in the Treatment of Endodontic Periapical Lesions in Adult Patients: A Narrative Review

by Agata Zoltowska, Katarzyna Machut, Elzbieta Pawlowska and Marcin Derwich

Pharmaceuticals 2021, 14(10), 1041; https://doi.org/10.3390/ph14101041 - 13 Oct 2021

Cited by 11 | Viewed by 4523

Abstract

Platelet concentrates have been widely used in regenerative medicine, including endodontics. The aim of this manuscript was to assess critically the efficacy of PRF in the treatment of endodontic periapical lesions in adult patients on the basis of the literature. The PICO approach [...] Read more.

Platelet concentrates have been widely used in regenerative medicine, including endodontics. The aim of this manuscript was to assess critically the efficacy of PRF in the treatment of endodontic periapical lesions in adult patients on the basis of the literature. The PICO approach was used to properly develop literature search strategies. The PubMed database was analyzed with the keywords: “((PRP) OR (PRF) OR (PRGF) OR (CGF)) AND (endodontic) AND ((treatment) OR (therapy))”. After screening of 155 results, 14 articles were included in this review. Different types of platelet concentrates are able to stimulate the processes of proliferation and differentiation of mesenchymal stem cells. Platelet rich fibrin (PRF) releases growth factors for at least 7 days at the application site. Growth factors and released cytokines stimulate the activity of osteoblasts. Moreover, the release of growth factors accelerates tissue regeneration by increasing the migration of fibroblasts. It was not possible to assess the efficacy of PRF supplementation in the treatment of endodontic periapical lesions in permanent, mature teeth with closed apexes, due to the lack of well-designed scientific research. Further studies are needed to analyze the effect of PRF on the healing processes in the periapical region. Full article

(This article belongs to the Section Pharmacology)

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19 pages, 3358 KiB

Open AccessArticle

Inhibition of 11?-HSD1 Ameliorates Cognition and Molecular Detrimental Changes after Chronic Mild Stress in SAMP8 Mice

by Dolors Puigoriol-Illamola, Júlia Companys-Alemany, Kris McGuire, Natalie Z. M. Homer, Rosana Leiva, Santiago Vázquez, Damian J. Mole, Christian Griñán-Ferré and Mercè Pallàs

Pharmaceuticals 2021, 14(10), 1040; https://doi.org/10.3390/ph14101040 - 13 Oct 2021

Cited by 6 | Viewed by 3239

Abstract

Impaired glucocorticoid (GC) signaling is a significant factor in aging, stress, and neurodegenerative diseases such as Alzheimer’s disease. Therefore, the study of GC-mediated stress responses to chronic moderately stressful situations, which occur in daily life, is of huge interest for the design of [...] Read more.

Impaired glucocorticoid (GC) signaling is a significant factor in aging, stress, and neurodegenerative diseases such as Alzheimer’s disease. Therefore, the study of GC-mediated stress responses to chronic moderately stressful situations, which occur in daily life, is of huge interest for the design of pharmacological strategies toward the prevention of neurodegeneration. To address this issue, SAMP8 mice were exposed to the chronic mild stress (CMS) paradigm for 4 weeks and treated with RL-118, an 11?-hydroxysteroid dehydrogenase type 1 (11?-HSD1) inhibitor. The inhibition of this enzyme is linked with a reduction in GC levels and cognitive improvement, while CMS exposure has been associated with reduced cognitive performance. The aim of this project was to assess whether RL-118 treatment could reverse the deleterious effects of CMS on cognition and behavioral abilities and to evaluate the molecular mechanisms that compromise healthy aging in SAMP8 mice. First, we confirmed the target engagement between RL-118 and 11?-HSD1. Additionally, we showed that DNA methylation, hydroxymethylation, and histone phosphorylation were decreased by CMS induction, and increased by RL-118 treatment. In addition, CMS exposure caused the accumulation of reactive oxygen species (ROS)-induced damage and increased pro-oxidant enzymes—as well as pro-inflammatory mediators—through the NF-?B pathway and astrogliosis markers, such as GFAP. Of note, these modifications were reversed by 11?-HSD1 inhibition. Remarkably, although CMS altered mTORC1 signaling, autophagy was increased in the SAMP8 RL-118-treated mice. We also showed an increase in amyloidogenic processes and a decrease in synaptic plasticity and neuronal remodeling markers in mice under CMS, which were consequently modified by RL-118 treatment. In conclusion, 11?-HSD1 inhibition through RL-118 ameliorated the detrimental effects induced by CMS, including epigenetic and cognitive disturbances, indicating that GC-excess attenuation shows potential as a therapeutic strategy for age-related cognitive decline and AD. Full article

(This article belongs to the Special Issue Stress, Neurotransmitters and Neurodegeneration)

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12 pages, 1149 KiB

Open AccessSystematic Review

Somatostatin Analogue Therapy in MEN1-Related Pancreatic Neuroendocrine Tumors from Evidence to Clinical Practice: A Systematic Review

by Anna La Salvia, Franz Sesti, Chiara Grinzato, Rossella Mazzilli, Maria Grazia Tarsitano, Elisa Giannetta and Antongiulio Faggiano

Pharmaceuticals 2021, 14(10), 1039; https://doi.org/10.3390/ph14101039 - 12 Oct 2021

Cited by 7 | Viewed by 3140

Abstract

Neuroendocrine neoplasms (NENs) are relatively rare and complex tumors that can be sporadic or hereditary, as in the context of multiple endocrine neoplasia type 1 (MEN1) where patients display a 70% lifelong risk of developing a pancreatic NENs (pNENs). To date, specific personalized [...] Read more.

Neuroendocrine neoplasms (NENs) are relatively rare and complex tumors that can be sporadic or hereditary, as in the context of multiple endocrine neoplasia type 1 (MEN1) where patients display a 70% lifelong risk of developing a pancreatic NENs (pNENs). To date, specific personalized treatment for pNENs in patients with MEN1 are lacking. The aim of this study was to systematically analyze the efficacy and safety of somatostatin analogue (SSA) treatment in patients affected by MEN1-related pNENs. We performed a systematic review of the literature, searching for peer-reviewed articles on SSA (octreotide or lanreotide) treatment in MEN1 associated with pNENs. We selected 20 studies with a pooled population of 105 MEN1 patients with pNENs. Females were 58.5%, median age was 44 years (18–73). TNM stage at diagnosis was stage I–II in 84.8% and stage IV in 15.2%. The overall response rate (SD+PR+CR) was achieved in 88.3% of cases, with stable disease in 75.6% and objective response in 12.7% of patients. The safety profile was favorable with both SSA agents. SSAs appear to be an effective and safe treatment option for MEN1-related pNEN, either at localized or advanced stages. Full article

(This article belongs to the Special Issue Neuroendocrine Neoplasms and Endocrine Glands Tumors: Pharmacological and Clinical Aspects)

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19 pages, 5042 KiB

Open AccessArticle

Engineered Bacteriophage as a Delivery Vehicle for Antibacterial Protein, SASP

by James Cass, Anne Barnard and Heather Fairhead

Pharmaceuticals 2021, 14(10), 1038; https://doi.org/10.3390/ph14101038 - 12 Oct 2021

Cited by 13 | Viewed by 3749

Abstract

The difficulties in developing novel classes of antibacterials is leading to a resurgence of interest in bacteriophages as therapeutic agents, and in particular engineered phages that can be optimally designed. Here, pre-clinical microbiology assessment is presented of a Staphylococcus aureus phage engineered to [...] Read more.

The difficulties in developing novel classes of antibacterials is leading to a resurgence of interest in bacteriophages as therapeutic agents, and in particular engineered phages that can be optimally designed. Here, pre-clinical microbiology assessment is presented of a Staphylococcus aureus phage engineered to deliver a gene encoding an antibacterial small acid soluble spore protein (SASP) and further, rendered non-lytic to give product SASPject PT1.2. PT1.2 has been developed initially for nasal decolonisation of S. aureus, including methicillin-resistant S. aureus. Time-kill curve assays were conducted with PT1.2 against a range of staphylococcal species, and serial passaging experiments were conducted to investigate the potential for resistance to develop. SASPject PT1.2 demonstrates activity against 100% of 225 geographically diverse S. aureus isolates, exquisite specificity for S. aureus, and a rapid speed of kill. The kinetics of S. aureus/PT1.2 interaction is examined together with demonstrating that PT1.2 activity is unaffected by the presence of human serum albumin. SASPject PT1.2 shows a low propensity for resistance to develop with no consistent shift in sensitivity in S. aureus cells passaged for up to 42 days. SASPject PT1.2 shows promise as a novel first-in-class antibacterial agent and demonstrates potential for the SASPject platform. Full article

(This article belongs to the Special Issue Bacteriophages as Therapeutic Delivery Vehicles)

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Mechanism of action of SASPject PT1.2 and SASP. Full article ">Figure 2
Modification of phi11 genome (A) showing holin and amidase genes (B) and creation of SASPject PT1.2 by deletion of holin gene and promoter driving holin and amidase genes by insertion of SASP-C gene under the control of native S. aureus promoter, fbaA and, selectable marker, cadmium resistance gene (C). Phi11 genome visualized using CGView [<a href="#B21-pharmaceuticals-14-01038" class="html-bibr">21</a>]. Full article ">Figure 3
Time-kill curve assay showing efficacy of SASPject PT1.2 in mixed S. aureus MSSA/MRSA (strains 100085/88048 respectively) cultures. Viable cells counts are shown for the strain written in bold letters. Results of a representative experiment are shown. Full article ">Figure 4
Time-kill curve assay showing efficacy of SASPject PT1.2 in mixed S. aureus MSSA/MRSA (strains 100085/17046 respectively) cultures. Viable cells counts are shown for the strain written in bold letters. Results of a representative experiment are shown. Full article ">Figure 5
Time-kill curve assay showing efficacy of SASPject PT1.2 in mixed cultures of MRSA strain 88048 and S. epidermidis 7,020 (abbreviated to “S.e.” in the chart legend). Viable cell counts are shown for the strain written in bold letters. Results of a representative experiment are shown. Full article ">Figure 6
Time-kill curve assay showing efficacy of SASPject PT1.2 in mixed cultures of MRSA and S. haemolyticus 133034 (abbreviated to “S.h.” in the chart legend). Viable cell counts are shown for the strain written in bold letters. Results of a representative experiment are shown. Full article ">Figure 7
Time-kill curve assay of SASPject PT1.2 activity against S. aureus USA 300 over 1 h. Results of a representative experiment are shown. Full article ">Figure 8
Time-kill curve analysis of SASPject PT1.2 activity, at concentrations ranging 106 to 108 pfu/mL, against EMRSA-15 strain 02ST4127 treated over 24 h. Data plotted are the means of 2 or 3 (107 pfu/mL only) replicates. Error bars represent 1 standard deviation. Full article ">Figure 9
Time-kill curve analysis of SASPject PT1.2 activity against EMRSA-15 strain 02ST4127, at viable cell concentrations of 103, 105, and 107 cfu/mL, over 6 h. The means of duplicate samples are plotted. Error bars represent 1 standard deviation. Full article ">Figure 10
Time-kill curve assay showing the efficacy of SASPject PT1.2 against S. aureus MRSA USA300 and EMRSA-16 in the presence and absence of human serum albumin (HSA) at 50 g/L. Data points represent the mean of 2 replicates and error bars represent 1 standard deviation. Full article ">Figure 11
Assessment of resistance development in serially passaged EMRSA strain O2ST4285 by time-kill curve assay of cells from passage day 52 (3 and 6 × 104 pfu/mL passaged cultures) and passage day 46 (1.5 × 104 pfu/mL passaged cultures) exposed to SASPject PT1.2. Passaging was carried out in triplicate, and time-kill curve assays were carried out in singlicate on each of the triplicate cultures. Cells that were passaged in singlicate in the absence of PT1.2 for 52 days were exposed to PT1.2 as a control (Control). The mean number of viable cells for each passaging condition following time-kill curve assay with PT1.2 is plotted. Error bars for the passaged conditions represent 1 standard deviation. Full article ">Figure 12
Assessment of resistance development in serially passaged MRSA USA300 by exposing passaged cells to PT1.2 at passage days 7, 14, and 21, and incubating with PT1.2 for 4 h before assessing viable cell count. Triplicate cultures were set up for each of the phage concentrations, and each culture was assessed via the kill experiment in singlicate. Results shown are the mean viable cell counts for the three cultures passaged with each PT1.2 concentration, following kill experiment analysis. Error bars represent 1 standard deviation. The control was MRSA USA300 simultaneously passaged for the same lengths of time in the absence of PT1.2, prior to assessment of viable cell count after exposure to PT1.2 for 4 h. Full article ">

17 pages, 2861 KiB

Open AccessArticle

Thermodynamic Study of Oxidovanadium(IV) with Kojic Acid Derivatives: A Multi-Technique Approach

by Rosita Cappai, Guido Crisponi, Daniele Sanna, Valeria Ugone, Andrea Melchior, Eugenio Garribba, Massimiliano Peana, Maria Antonietta Zoroddu and Valeria Marina Nurchi

Pharmaceuticals 2021, 14(10), 1037; https://doi.org/10.3390/ph14101037 - 12 Oct 2021

Cited by 6 | Viewed by 2612

Abstract

The good chelating properties of hydroxypyrone (HPO) derivatives towards oxidovanadium(IV) cation, V^IVO²⁺, constitute the precondition for the development of new insulin-mimetic and anticancer compounds. In the present work, we examined the V^IVO²⁺ complex formation equilibria of [...] Read more.

The good chelating properties of hydroxypyrone (HPO) derivatives towards oxidovanadium(IV) cation, V^IVO²⁺, constitute the precondition for the development of new insulin-mimetic and anticancer compounds. In the present work, we examined the V^IVO²⁺ complex formation equilibria of two kojic acid (KA) derivatives, L4 and L9, structurally constituted by two kojic acid units linked in position 6 through methylene diamine and diethyl-ethylenediamine, respectively. These chemical systems have been characterized in solution by the combined use of various complementary techniques, as UV-vis spectrophotometry, potentiometry, NMR and EPR spectroscopy, ESI-MS spectrometry, and DFT calculations. The thermodynamic approach allowed proposing a chemical coordination model and the calculation of the complex formation constants. Both ligands L4 and L9 form 1:1 binuclear complexes at acidic and physiological pHs, with various protonation degrees in which two KA units coordinate each V^IVO²⁺ ion. The joined use of different techniques allowed reaching a coherent vision of the complexation models of the two ligands toward oxidovanadium(IV) ion in aqueous solution. The high stability of the formed species and the binuclear structure may favor their biological action, and represent a good starting point toward the design of new pharmacologically active vanadium species. Full article

(This article belongs to the Special Issue Applications of Medicinal Bioinorganic Chemistry)

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10 pages, 1097 KiB

Open AccessArticle

Folium Sennae Increased the Bioavailability of Methotrexate through Modulation on MRP 2 and BCRP

by Chung-Ping Yu, Yu-Hsuan Peng, Ching-Ya Huang, Yow-Wen Hsieh, Yu-Chi Hou and Shiuan-Pey Lin

Pharmaceuticals 2021, 14(10), 1036; https://doi.org/10.3390/ph14101036 - 12 Oct 2021

Cited by 3 | Viewed by 2393

Abstract

Folium Sennae (FS), a popular laxative (Senna), contains polyphenolic anthranoids, whose conjugation metabolites are probable modulators of multidrug resistance-associated proteins (MRPs) and breast cancer resistance protein (BCRP). We suspected that the combined use of FS might alter the pharmacokinetics of various medicines transported [...] Read more.

Folium Sennae (FS), a popular laxative (Senna), contains polyphenolic anthranoids, whose conjugation metabolites are probable modulators of multidrug resistance-associated proteins (MRPs) and breast cancer resistance protein (BCRP). We suspected that the combined use of FS might alter the pharmacokinetics of various medicines transported by MRPs or BCRP. This study investigated the effect of FS on the pharmacokinetics of methotrexate (MTX), an anticancer drug and a probe substrate of MRPs/BCRP. Rats were orally administered MTX alone and with two dosage regimens of FS in a parallel design. The results show that 5.0 g/kg of FS significantly increased the AUC_0–2880, AUC_720–2880 and MRT of MTX by 45%, 102% and 42%, and the seventh dose of 2.5 g/kg of FS significantly enhanced the AUC_720–2880 and MRT by 78% and 42%, respectively. Mechanism studies indicated that the metabolites of FS (FSM) inhibited MRP 2 and BCRP. In conclusion, the combined use of FS increased the systemic exposure and MRT of MTX through inhibition on MRP 2 and BCRP. Full article

(This article belongs to the Section Pharmacology)

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20 pages, 13804 KiB

Open AccessArticle

Growth Inhibition of Triple-Negative Breast Cancer: The Role of Spatiotemporal Delivery of Neoadjuvant Doxorubicin and Cisplatin

by Dominick Salerno and Stavroula Sofou

Pharmaceuticals 2021, 14(10), 1035; https://doi.org/10.3390/ph14101035 - 12 Oct 2021

Cited by 2 | Viewed by 2669

Abstract

Combinations of platinum-based compounds with doxorubicin in free and/or in liposomal form for improved safety are currently being evaluated in the neoadjuvant setting on patients with advanced triple-negative breast cancer (TNBC). However, TNBC may likely be driven by chemotherapy-resistant cells. Additionally, established TNBC [...] Read more.

Combinations of platinum-based compounds with doxorubicin in free and/or in liposomal form for improved safety are currently being evaluated in the neoadjuvant setting on patients with advanced triple-negative breast cancer (TNBC). However, TNBC may likely be driven by chemotherapy-resistant cells. Additionally, established TNBC tumors may also exhibit diffusion-limited transport, resulting in heterogeneous intratumoral delivery of the administered therapeutics; this limits therapeutic efficacy in vivo. We studied TNBC cells with variable chemosensitivities, in the absence (on monolayers) and presence (in 3D multicellular spheroids) of transport barriers; we compared the combined killing effect of free doxorubicin and free cisplatin to the killing effect (1) of conventional liposomal forms of the two chemotherapeutics, and (2) of tumor-responsive lipid nanoparticles (NP), specifically engineered to result in more uniform spatiotemporal microdistributions of the agents within solid tumors. This was enabled by the NP properties of interstitial release, cell binding/internalization, and/or adhesion to the tumors’ extracellular matrix. The synergistic cell kill by combinations of the agents (in all forms), compared to the killing effect of each agent alone, was validated on monolayers of cells. Especially for spheroids formed by cells exhibiting resistance to doxorubicin combination treatments with both agents in free and/or in tumor-responsive NP-forms were comparably effective; we not only observed greater inhibition of outgrowth compared to the single agent(s) but also compared to the conventional liposome forms of the combined agents. We correlated this finding to more uniform spatiotemporal microdistributions of agents by the tumor-responsive NP. Our study shows that combinations of NP with properties specifically optimized to improve the spatiotemporal uniformity of the delivery of their corresponding therapeutic cargo can improve treatment efficacy while keeping favorable safety profiles. Full article

(This article belongs to the Special Issue Potential Molecular Targets and Therapeutics in Triple-Negative Breast Cancer)

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11 pages, 1994 KiB

Open AccessArticle

The Extract of Corydalis yanhusuo Prevents Morphine Tolerance and Dependence

by Lamees Alhassen, Khawla Nuseir, Allyssa Ha, Warren Phan, Ilias Marmouzi, Shalini Shah and Olivier Civelli

Pharmaceuticals 2021, 14(10), 1034; https://doi.org/10.3390/ph14101034 - 12 Oct 2021

Cited by 9 | Viewed by 9833

Abstract

The opioid epidemic was triggered by an overprescription of opioid analgesics. In the treatment of chronic pain, repeated opioid administrations are required which ultimately lead to tolerance, physical dependence, and addiction. A possible way to overcome this conundrum consists of a co-medication that [...] Read more.

The opioid epidemic was triggered by an overprescription of opioid analgesics. In the treatment of chronic pain, repeated opioid administrations are required which ultimately lead to tolerance, physical dependence, and addiction. A possible way to overcome this conundrum consists of a co-medication that maintains the analgesic benefits of opioids while preventing their adverse liabilities. YHS, the extract of the plant Corydalis yanhusuo, has been used as analgesic in traditional Chinese medicine for centuries. More recently, it has been shown to promote analgesia in animal models of acute, inflammatory, and neuropathic pain. It acts, at least in part, by inhibiting the dopamine D2 receptor, suggesting that it may be advantageous to manage addiction. We first show that, in animals, YHS can increase the efficacy of morphine antinociceptive and, as such, decrease the need of the opioid. We then show that YHS, when coadministered with morphine, inhibits morphine tolerance, dependence, and addiction. Finally, we show that, in animals treated for several days with morphine, YHS can reverse morphine dependence and addiction. Together, these data indicate that YHS may be useful as a co-medication in morphine therapies to limit adverse morphine effects. Because YHS is readily available and safe, it may have an immediate positive impact to curb the opioid epidemic. Full article

(This article belongs to the Section Natural Products)

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