Pharmaceuticals

15 pages, 4425 KiB

Open AccessArticle

Discovery of Substituted (2-Aminooxazol-4-yl)Isoxazole-3-carboxylic Acids as Inhibitors of Bacterial Serine Acetyltransferase in the Quest for Novel Potential Antibacterial Adjuvants

by Joana Magalhães, Nina Franko, Samanta Raboni, Giannamaria Annunziato, Päivi Tammela, Agostino Bruno, Stefano Bettati, Stefano Armao, Costanza Spadini, Clotilde Silvia Cabassi, Andrea Mozzarelli, Marco Pieroni, Barbara Campanini and Gabriele Costantino

Pharmaceuticals 2021, 14(2), 174; https://doi.org/10.3390/ph14020174 - 23 Feb 2021

Cited by 7 | Viewed by 2933

Abstract

Many bacteria and actinomycetales use L-cysteine biosynthesis to increase their tolerance to antibacterial treatment and establish a long-lasting infection. In turn, this might lead to the onset of antimicrobial resistance that currently represents one of the most menacing threats to public health worldwide. [...] Read more.

Many bacteria and actinomycetales use L-cysteine biosynthesis to increase their tolerance to antibacterial treatment and establish a long-lasting infection. In turn, this might lead to the onset of antimicrobial resistance that currently represents one of the most menacing threats to public health worldwide. The biosynthetic machinery required to synthesise L-cysteine is absent in mammals; therefore, its exploitation as a drug target is particularly promising. In this article, we report a series of inhibitors of Salmonella thyphimurium serine acetyltransferase (SAT), the enzyme that catalyzes the rate-limiting step of L-cysteine biosynthesis. The development of such inhibitors started with the virtual screening of an in-house library of compounds that led to the selection of seven structurally unrelated hit derivatives. A set of molecules structurally related to hit compound 5, coming either from the original library or from medicinal chemistry efforts, were tested to determine a preliminary structure–activity relationship and, especially, to improve the inhibitory potency of the derivatives, that was indeed ameliorated by several folds compared to hit compound 5 Despite these progresses, at this stage, the most promising compound failed to interfere with bacterial growth when tested on a Gram-negative model organism, anticipating the need for further research efforts. Full article

(This article belongs to the Special Issue Small Molecules as Antimicrobials)

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Figure 1

Figure 1
Schematic representation of the reductive sulfate assimilation pathway in Salmonella. Inorganic sulfate is transported inside the cell where it is reduced to bisulfide by a multi-step process. L-serine is activated by O-acetylation by SAT to give the product OAS, which spontaneously tautomerizes to NAS, the inducer of the cysteine regulon. A β-substitution reaction between OAS and HS-, catalyzed by OASS, leads to L-cysteine formation. This amino acid and the pathways of its biosynthesis influence many processes relevant to infection, like resistance to oxidative stress, biofilm formation and antibiotic resistance. UPAR415 is a competitive inhibitor of OASS. Full article ">Figure 2
Compounds from the virtual screening. Full article ">Figure 3
Inhibitory activity of compound 5. Panel A: dependence of relative initial reaction rate on the concentration of compound 5. The line through data points is the fitting to equation 1 with IC50 = 110 ± 0 μM. Panel B: dependence of the initial reaction rate on the concentration of acetylCoA in the absence of added inhibitors (grey dots) and in the presence of different concentrations of compound 5 (green dots 30 μM; pink dots 100 μM, red dots 300 μM). The series of lines through data points is the fitting to the linearised form of equation 2 with Ki = 64 ± 12 μM. Full article ">Figure 4
(A) docking pose of compound 5 in AcCoA pocket in EcSAT (alignment of structures with pdb code 1T3D and pdb code 1SSM). (B) Overlay of the docking pose of compound 5 and AcCoA structure. Full article ">Scheme 1
a Reagent and conditions: (a) triethylamine, tetrahydrofuran, rt; (b) N-Bromosuccinimide, p-toluenesulfonic acid, acetonitrile, reflux; (c) urea, dimethylformamide, MW (120 °C, 300 W), 3 min, 49%; (d) suitable bromobenzene derivative or 4-bromopyridine (21e), Sodium t-butoxide, X-Phos Pd G2, t-butanol, toluene, MW (130 °C, 300 W), 15 min, 8–35%; (e) LiOH, THF/H2O/MeOH 3:1:1, rt, 3 h, quantitative. Full article ">

16 pages, 2360 KiB

Open AccessArticle

Preclinical Pharmacokinetics and Biodistribution of Anticancer Dinuclear Palladium(II)-Spermine Complex (Pd₂Spm) in Mice

by Martin Vojtek, Salomé Gonçalves-Monteiro, Edgar Pinto, Sára Kalivodová, Agostinho Almeida, Maria P. M. Marques, Ana L. M. Batista de Carvalho, Clara B. Martins, Helder Mota-Filipe, Isabel M. P. L. V. O. Ferreira and Carmen Diniz

Pharmaceuticals 2021, 14(2), 173; https://doi.org/10.3390/ph14020173 - 23 Feb 2021

Cited by 14 | Viewed by 3784

Abstract

Palladium-based compounds are regarded as potential analogs to platinum anticancer drugs with improved properties. The present study assessed the pharmacokinetics and biodistribution of a dinuclear palladium(II)-spermine chelate (Pd₂Spm), which has previously been shown to possess promising in vitro activity against several [...] Read more.

Palladium-based compounds are regarded as potential analogs to platinum anticancer drugs with improved properties. The present study assessed the pharmacokinetics and biodistribution of a dinuclear palladium(II)-spermine chelate (Pd₂Spm), which has previously been shown to possess promising in vitro activity against several therapy-resistant cancers. Using inductively coupled plasma-mass spectrometry, the kinetic profiles of palladium/platinum in serum, serum ultrafiltrate and tissues (kidney, liver, brain, heart, lungs, ovaries, adipose tissue and mammary glands) were studied in healthy female Balb/c mice after a single intraperitoneal bolus injection of Pd₂Spm (3 mg/kg bw) or cisplatin (3.5 mg/kg bw) between 0.5 and 48 h post-injection. Palladium in serum exhibited biphasic kinetics with a terminal half-life of 20.7 h, while the free palladium in serum ultrafiltrate showed a higher terminal half-life than platinum (35.5 versus 31.5 h). Palladium was distributed throughout most of the tissues except for the brain, with the highest values in the kidney, followed by the liver, lungs, ovaries, adipose tissue and mammary glands. The in vitro cellular accumulation was also evaluated in breast cancer cells, evidencing a passive diffusion as a mechanism of Pd₂Spm’s cellular entry. This study reports, for the first time, the favorable pharmacokinetics and biodistribution of Pd₂Spm, which may become a promising pharmacological agent for cancer treatment. Full article

(This article belongs to the Special Issue Metal-Based Drugs: Updates and Perspectives)

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Graphical abstract

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Full article ">Figure 1
Structure of Pt(II)- and Pd(II)-based agents: Worldwide approved Pt drugs—(a) cisplatin, (b) carboplatin and (c) oxaliplatin. (d) Novel palladium(II)-based complex Pd2Spm. Full article ">Figure 2
Kinetic profiles of Pd (from Pd2Spm, 3 mg/kg bw) and Pt (from cisplatin, 3.5 mg/kg bw) in serum and serum ultrafiltrate after single intraperitoneal bolus injection in Balb/c mice. Data are expressed as mean ± SEM (n = 5 animals per time point). (a) Concentration–time plot of mean Pd or Pt levels in serum and serum ultrafiltrate. (b) Concentration–time plot of mean Pd or Pt levels in serum and serum ultrafiltrate normalized to administered metal dose. Full article ">Figure 3
Drug binding to plasma proteins versus time, for cisplatin and Pd2Spm. Data are expressed as mean ± SEM (n = 5 animals per time point) and were compared with Student’s t-test. * p < 0.05, ** p < 0.01, *** p < 0.001. Full article ">Figure 4
Biodistribution of total Pd (from Pd2Spm, 3 mg/kg bw) and Pt (from cisplatin, 3.5 mg/kg bw) in tissues after single intraperitoneal bolus injection in Balb/c mice. Data are expressed as mean ± SEM (n = 5 animals per time point) and were compared with Student’s t-test. * p < 0.05, ** p < 0.01, *** p < 0.001. (a) AUC0-48h (area under the Pd/Pt concentration–time curve). Units: h × ng/g of tissue; (b) dose-normalized AUC0–48 h (area under the Pd/Pt concentration–time curve). Units: h × ng/g per µg metal dose. Full article ">Figure 5
Cellular accumulation of Pd2Spm and cisplatin in (a) MDA-MB-231 and (b) HCC-1143 triple-negative breast cancer cells. Data are expressed as mean ± SEM (n = 4 independent experiments) and were compared with Student’s t-test. ** p < 0.01, *** p < 0.001. Full article ">Figure A1
Kinetic profiles of Pd (from Pd2Spm, 3 mg/kg bw) and Pt (from cisplatin, 3.5 mg/kg bw) in tissues after single intraperitoneal bolus injection in Balb/c mice. Data are expressed as mean ± SEM (n = 5 animals per time point). Units: h.ng/g of tissue (a) Concentration–time plot of mean Pd levels in tissues. (b) Concen-tration–time plot of mean Pt levels in tissues. Full article ">Figure A2
Metal dose-normalized kinetic profiles of Pd (from Pd2Spm, 3 mg/kg bw) and Pt (from cisplatin, 3.5 mg/kg bw) in tissues after single intraperitoneal bolus injection in Balb/c mice. Data are expressed as mean ± SEM (n = 5 animals per time point). Units: h.ng/g per µg metal dose (a) Concentration–time plot of mean Pd levels (normalized to administered metal dose) in tissues. (b) Concentration–time plot of mean Pt levels (normalized to administered metal dose) in tissues. Full article ">

12 pages, 2587 KiB

Open AccessArticle

Up-Regulated Vitamin D Receptor by Pelargonium sidoides Extract EPs^® 7630 Contributes to Rhinovirus Defense in Bronchial Epithelial Cells

by Michael Roth, Qingzhu Sun and Michael Tamm

Pharmaceuticals 2021, 14(2), 172; https://doi.org/10.3390/ph14020172 - 22 Feb 2021

Cited by 13 | Viewed by 3375

Abstract

EPs^®7630, extracted from Pelargonium sidoides, reduces the severity of viral upper respiratory tract infections. Vitamin D also improves anti-viral host defense through similar signaling pathways. This study assessed if EPs^®7630 modifies vitamin D receptor (VDR) expression and function [...] Read more.

EPs^®7630, extracted from Pelargonium sidoides, reduces the severity of viral upper respiratory tract infections. Vitamin D also improves anti-viral host defense through similar signaling pathways. This study assessed if EPs^®7630 modifies vitamin D receptor (VDR) expression and function by human bronchial epithelial cells. Bronchial epithelial cells were incubated with EPs^®7630 over 48 h before calcitriol stimulation and/or infection with Rhinovirus (RV)-16. Protein expression was determined by Western-blotting. Intracellular signaling of mitogen activated protein kinases (MAPK) was studied by chemical inhibitors. The anti-viral effect was assessed by immunofluorescence for RV-16 protein. EPs^®7630 upregulated VDR expression through Erk1/2 MAPK and thereby increased the cell’s sensitivity to calcitriol. Compared ton untreated cells, the shift of the VDR into the nucleus at 5.3 times lower calcitriol concentration. EPs^®7630 increased Erk1/2 MAPK signaling, but reduced p38 phosphorylation, and had no effect on Jun N-terminal kinase (JNK). EPs^®7630 improved the anti-viral effect of vitamin D on RV-16 infection by 2.1 folds compared to vitamin D alone or to untreated cells. Furthermore, EPs^®7630 improved the differentiation of epithelial cells by upregulating E-cadherin expression through Erk1/2. In conclusion, EPs^®7630 increased host defense against Rhinovirus infection by upregulating the VDR and the differentiation of epithelial cells. Full article

(This article belongs to the Special Issue Natural Pharmacons: Biologically Active Plant Based Pharmaceuticals)

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Graphical abstract

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Full article ">Figure 1
High pressure liquid chromatography (HPLC) fingerprint detected by UV at 280 nm. The major peaks were assigned by analysis of high resolution mass spectrometry (HRMS) data and are given as follows: (a) adenosine 3′,5′-cyclic monophosphate, (b) guanosine 3′,5′-cyclic monophosphate, (c) 1-methylguanosine 3′,5′-cyclic monophosphate, and (d) benzopyranones. The assigned structures are given in <a href="#pharmaceuticals-14-00172-t001" class="html-table">Table 1</a>. Full article ">Figure 2
EPs® 7630 modifies vitamin D receptor (VDR) expression. (A) Representative Western-blots and image analyses of VDR expression in primary bronchial epithelial cells (n = 4) stimulated with increasing concentrations of EPs® 7630 over 24 h. Bars represent mean ± SEM. (B) Kinetic of EPs® 7630 (10 μg/mL) induced VDR expression over 48 h (n = 4). (C) The effect of EPs® 7630 on the phosphorylation of Erk1/2 mitogen activated protein kinases (MAPK) over 120 min. (D) The effect of inhibitors for Jun N-terminal kinase (JNK), p38, and Erk1/2 on EPs® 7630 induced VDR expression (n = 4). Representative Western-blots are depicted above the corresponding bars. Similar results were obtained in BEAS-2B cells. Full article ">Figure 3
EPs® 7630 and calcitriol activate the VDR. (A) Concentration dependent increase of nuclear VDR by EPs® 7630 over 24 h in primary human bronchial epithelial cells (n = 4). (B) Concentration dependent increase of nuclear VDR by calcitriol over 24 h (n = 4). (C) The effect of combined EPs® 7630 with calcitriol on the ratio of cytosolic versus nuclear VDR accumulation (n = 4) over 24 h. (D) The effect of MAPK inhibitors for JNK (SP600125), p38 (SB203580), and Erk1/2 (PD98059) on EPs® 7630 and calcitriol induced VDR expression (n = 3). Bars represent mean ± SEM. Similar results were obtained in BEAS-2B cells. Full article ">Figure 4
The effect of EPs® 7630 and calcitriol on RV-16 infection in BEAS-2B cells. (A) Concentration dependent effect of EPs® 7630 on RV-16 staining in BEAS-2B cells. (B) Concentration dependent effect of calcitriol on RV-16 staining in BEAS-2B cells. (C) The effect of combined EPs® 7630 (fixed concentration) with calcitriol (increasing concentrations) on RV-16 staining on BEAS-2B cells. Bars represent mean ± SEM of triplicate experiments. (D) Representative immunofluorescence photographs of RV-16 positive primary epithelial cells in the presence and absence of EPs® 7630 (1 µg/mL), or calcitriol (1 µM), or the combination of both (EPs® 7630 1 µg/mL + calcitriol 1 µM). Full article ">Figure 5
EPs® 7630 increases E-cadherin expression through Erk1/2 MAPK in BEAS-2B cells. (A) Concentration effect of EPs® 7630 on the expression of E-cadherin over 24 h in BEAS-2B cells. (B) The effect of inhibitors for JNK, p38, and Erk1/2 on EPs® 7630 induced E-cadherin expression. Bars represent mean ± SEM of triplicate experiments. Full article ">Figure 6
Cell characterization and treatment. (A) Cell type characterization by phase contrast microscopy and positive immunofluorescence pan-cytokeratin, E-cadherin, and fibronectin (negative control). (B) Treatment scheme of cells. Confluent epithelial cells were pre-treated with either EPs® 7630 for 24 h before calcitriol was added for another 24 h and prior to infection with 1 MOI RV-16. RNA and protein were isolated over 4 days. Non-infected cells were used to calculate changes of protein expression at the corresponding time points. Full article ">

13 pages, 949 KiB

Open AccessArticle

Cannabis-Based Oral Formulations for Medical Purposes: Preparation, Quality and Stability

by Francesca Baratta, Marco Simiele, Irene Pignata, Lorenzo Ravetto Enri, Antonio D’Avolio, Riccardo Torta, Anna De Luca, Massimo Collino and Paola Brusa

Pharmaceuticals 2021, 14(2), 171; https://doi.org/10.3390/ph14020171 - 22 Feb 2021

Cited by 13 | Viewed by 5969

Abstract

Current legislation in Italy provides that medical Cannabis may be administered orally or by inhalation. One of the fundamental criteria for the administration of oral formulations is that they deliver a known consistent quantity of the active ingredients to ensure uniform therapies leading [...] Read more.

Current legislation in Italy provides that medical Cannabis may be administered orally or by inhalation. One of the fundamental criteria for the administration of oral formulations is that they deliver a known consistent quantity of the active ingredients to ensure uniform therapies leading to the optimisation of the risks/benefits. In 2018, our group developed an improved Cannabis oil extraction technique. The objective of the present work was to carry out a stability study for the oil extracts obtained by this method. Furthermore, in order to facilitate the consumption of the prescribed medical Cannabis therapy by patients, a standard procedure was defined for the preparation of a single-dose preparation for oral use (hard capsules) containing the oil extract; thereafter, the quality and stability were evaluated. The hard capsules loaded with the oil extract were analysed and found to be uniform in content. The encapsulation process did not alter the quantity of the active molecule present in the oil. The stability tests yielded excellent results. Since the capsule dosage form is easily transported and administered, has pleasant organoleptic properties and is stable at room temperature for extended periods of time, this would facilitate the adherence to therapy by patients in treatment. Full article

(This article belongs to the Special Issue Clinical and Forensic Toxicology: The Latest Updates)

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15 pages, 1773 KiB

Open AccessCommunication

PARP Traps Rescue the Pro-Inflammatory Response of Human Macrophages in the In Vitro Model of LPS-Induced Tolerance

by Julita Pietrzak, Karolina Gronkowska and Agnieszka Robaszkiewicz

Pharmaceuticals 2021, 14(2), 170; https://doi.org/10.3390/ph14020170 - 22 Feb 2021

Cited by 3 | Viewed by 3178

Abstract

Secondary infections cause sepsis that lead to patient disability or death. Contact of macrophages with bacterial components (such as lipopolysaccharide—LPS) activates the intracellular signaling pathway downstream of Toll-like receptors (TLR), which initiate an immune proinflammatory response. However, the expression of nuclear factor-kappa B [...] Read more.

Secondary infections cause sepsis that lead to patient disability or death. Contact of macrophages with bacterial components (such as lipopolysaccharide—LPS) activates the intracellular signaling pathway downstream of Toll-like receptors (TLR), which initiate an immune proinflammatory response. However, the expression of nuclear factor-kappa B (NF-κB)-dependent proinflammatory cytokines significantly decreases after single high or multiple LPS stimulations. Knowing that poly(ADP-ribose) polymerase-1 (PARP1) serves as a cofactor of NF-κB, we aimed to verify a hypothesis of the possible contribution of PARP1 to the development of LPS-induced tolerance in human macrophages. Using TNF-α mRNA expression as a readout, we demonstrate that PARP1 interaction with the TNF-α promoter, controls macrophage immunoparalysis. We confirm that PARP1 is extruded from the gene promoter, whereas cell pretreatment with Olaparib maintains macrophage responsiveness to another LPS treatment. Furthermore, cell pretreatment with proteasome inhibitor MG132 completely abrogates the effect of Olaparib, suggesting that PARP1 acts with NF-κB in the same regulatory pathway, which controls pro-inflammatory cytokine transcription. Mechanistically, PARP1 trapping allows for the re-rebinding of p65 to the TNF-α promoter in LPS-stimulated cells. In conclusion, PARP traps prevent PARP1 extrusion from the TNF-α promoter upon macrophage stimulation, thereby maintaining chromatin responsiveness of TLR activation, allowing for the re-binding of p65 and TNF-α transcription. Full article

(This article belongs to the Special Issue Epigenetic Drugs)

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Figure 1
PARP traps maintain proinflammatory response in human macrophages activated with tolerance-inducing LPS doses. (a) The scheme of tolerance induction presents an experimental approach and cell treatments: the first dose of LPS aims to induce cell paralysis or priming within 24 h, the second serves to check macrophage pro-inflammatory response using mRNA of TNF-α as a readout after cell stimulation for 2 h. To test a possible effect of the key enzymes involved in the ADP-ribosylation metabolism (PARP inhibitors: olaparib, veliparib, niraparib, and PARG inhibitor: ADP–HPD) on the tolerance development, the corresponding compounds were added for 1 h prior to the tolerance-inducing (first) dose of bacterial endotoxin. Expression of TNF-α was quantified by real-time PCR, normalized to median of ACTB and GAPDH, and shown as a fold change with respect to control cells (LPS untreated = 1). (b) Three doses of LPS were tested for macrophage paralysis and the immunomodulatory effect of PARP inhibitor—olaparib (1 µM) was estimated based on the TNF-α transcription. The red rectangular indicates the couple: olaparib-LPS that was chosen for further experiments. (c) The other two PARP inhibitors, which differ in PARP-DNA binding ability—niraparib (MK-4827) and veliparib (ABT-888), as well as (d–e) PARG inhibitor—ADP–HPD (10 µM) were analyzed for their possible effect on the induction of tolerance by LPS. (d) The increased accumulation of ADP-ribosylated proteins caused by ADP–HPD pretreatment of LPS-induced macrophages was confirmed by western blot. H3 was used as a loading control. Full-length western blot images are included in the <a href="#app1-pharmaceuticals-14-00170" class="html-app">Supplementary Figure S3</a>. (e) The possible modulatory role of PARP1 in the paralysis of macrophage pro-inflammatory phenotype was tested also for other cytokine genes, such as IL-1β, IL-6, IL-12, MIP2A, COX2, and iNOS using real-time PCR for the measurement of mRNA levels. Bars in the figures represent mean ± standard error of the mean (SEM). One-way analysis of variance (ANOVA1) was carried out in GraphPad Prism 5 to compare means in several groups. Once the significance was detected, ANOVA1 was followed by the Tukey post-hoc test and significant differences between the two considered means are marked with * when significant at p < 0.05, ns—non-significant at p > 0.05. Abbreviations: iPARP—poly(ADP-ribose) polymerase (PARP) inhibitor(s), iPARG—poly(ADP-ribose) glycohydrolase (PARG) inhibitor, LPS—lipopolysaccharide, Olap—olaparib, Velip—veliparib, Nirap—niraparib, TNF—tumor necrosis factor, IL1β—interleukin 1 beta, IL6—interleukin 6, IL-12A—interleukin 12 subunit alpha, COX2—cyclooxygenase-2, MIP2A—macrophage inflammatory protein 2 subunit alpha, iNOS—inducible nitric oxide synthase, ACTB—actin beta, GAPDH—glyceraldehyde 3-phosphate dehydrogenase. Full article ">Figure 2
The PARP1 level drives LPS-induced immune paralysis in THP1 cells. (a) The induction of tolerance in acute leukemia cells was evaluated based on the TNF-α expression that was quantified by real-time PCR, normalized as median of ACTB and GAPDH, and shown as a fold change with respect to untreated cells. (b) The illustration in outlines the human proximal TNF-α promoter and the position of NF-κB (−98) as well as primer (forward: −124, reverse: −73) binding sites with respect to transcriptional start site (TSS). The fragment between the two indicated primers was amplified to quantify the immunoprecipitated DNA by real-time PCR. (c) The impact of the single LPS dose and olaparib on PARP1 occurrence at the TNF-α promoter was estimated with ChIP-qPCR. (d–e) The silencing efficiency of PARP1 in THP1 cells stably transfected with PARP-1 shRNA Plasmid was determined by real-time PCR and western blot, respectively versus corresponding control cells (shCTRL; control shRNA Plasmid-A). PARP1 mRNA was normalized to the median of ACTB and GAPDH, and shown as a fold change with respect to control cells with the normal PARP1 expression. In western blot, H3 was used as a loading control. Full-length western blot images are available in the <a href="#app1-pharmaceuticals-14-00170" class="html-app">Supplementary Figure S4</a>. (f) The mRNA level of TNF-α, MIP2a, and COX2, which served to assess the development of immune tolerance in response to LPS, was measured with real-time PCR as described above. (g–h) PARP1 overexpression in pCMV3–PARP1 versus pCMV3–EMPTY-transfected cells was confirmed by real-time PCR and western blot, respectively. Full-length western blot images are included in the <a href="#app1-pharmaceuticals-14-00170" class="html-app">Supplementary Figure S5</a>. mRNA level of PARP1 was normalized to ACTB and GAPDH and is presented as a fold change of PARP1 knock-in with respect to control cells. (i) The effect of PARP poison (olaparib) on the development of endotoxin tolerance in pCMV3–EMPTY and pCMV3–PARP1 cells was estimated on the basis of TNF-α transcription (mRNA), which was set as a readout. Bars in the figures represent mean ± standard error of the mean (SEM). One-way analysis of variance (ANOVA1) was carried out in GraphPad Prism 5 to compare means in several groups. Once the significance was detected, ANOVA1 was followed by the Tukey post-hoc test and significant differences between the two considered means are marked with * when significant at p < 0.05, ** when significant at p < 0.01, *** when significant at p < 0.001, ns—non-significant at p > 0.05. Abbreviations: THP1—human monocytic cell line, LPS—lipopolysaccharide, Olap—Olaparib, TNF—tumor necrosis factor, ACTB—actin beta, GAPDH—glyceraldehyde 3-phosphate dehydrogenase, shCTRL—THP1 cell transfected with Control shRNA Plasmid-A, shPARP1—THP1 cell transfected with PARP-1 shRNA Plasmid, pCMV3–PARP1—THP1 cells transfected with pCMV3–-PARP1 plasmid, pCMV3–EMPTY—THP1 cells transfected with pCMV3-EMPTY plasmid, IgG—immunoglobulin G. Full article ">Figure 3
PARP1 eviction from the TNF-α promoter prevents p65 re-binding after LPS re-stimulation. (A) The contribution of canonical NF-κB to macrophage response to LPS was estimated by comparing TNF-α transcription in control and cells pre-treated with proteasome inhibitor (MG132; 1 µM). mRNA level of the cytokine was quantified by real-time PCR, normalized to median of ACTB and GAPDH and presented as a fold change with respect to untreated cells. (B) p65 and (C) p50 occurrence at the TNF-α promoter was analyzed by ChIP-real-time PCR. Bars in the figures represent mean ± standard error of the mean (SEM). One-way analysis of variance (ANOVA1) was carried out in GraphPad Prism 5 to compare means in several groups. Once the significance was detected, ANOVA1 was followed by the Tukey post-hoc test and significant differences between the two considered means are marked with * when significant at p < 0.05, ** when significant at p < 0.01, ns—non-significant at p > 0.05. Abbreviations: LPS—lipopolysaccharide, Olap—olaparib, TNF—tumor necrosis factor, ACTB—actin beta, GAPDH—glyceraldehyde 3-phosphate dehydrogenase, IgG—immunoglobulin G, p65—transcription factor p65 (RelA), p50—nuclear factor NF-kappa-B p105 subunit. Full article ">

54 pages, 14187 KiB

Open AccessArticle

Synthesis and Biological Evaluation of 1-(Diarylmethyl)-1H-1,2,4-triazoles and 1-(Diarylmethyl)-1H-imidazoles as a Novel Class of Anti-Mitotic Agent for Activity in Breast Cancer

by Gloria Ana, Patrick M. Kelly, Azizah M. Malebari, Sara Noorani, Seema M. Nathwani, Brendan Twamley, Darren Fayne, Niamh M. O’Boyle, Daniela M. Zisterer, Elisangela Flavia Pimentel, Denise Coutinho Endringer and Mary J. Meegan

Pharmaceuticals 2021, 14(2), 169; https://doi.org/10.3390/ph14020169 - 22 Feb 2021

Cited by 6 | Viewed by 5240

Abstract

We report the synthesis and biochemical evaluation of compounds that are designed as hybrids of the microtubule targeting benzophenone phenstatin and the aromatase inhibitor letrozole. A preliminary screening in estrogen receptor (ER)-positive MCF-7 breast cancer cells identified 5-((2H-1,2,3-triazol-1-yl)(3,4,5-trimethoxyphenyl)methyl)-2-methoxyphenol 24 as a [...] Read more.

We report the synthesis and biochemical evaluation of compounds that are designed as hybrids of the microtubule targeting benzophenone phenstatin and the aromatase inhibitor letrozole. A preliminary screening in estrogen receptor (ER)-positive MCF-7 breast cancer cells identified 5-((2H-1,2,3-triazol-1-yl)(3,4,5-trimethoxyphenyl)methyl)-2-methoxyphenol 24 as a potent antiproliferative compound with an IC₅₀ value of 52 nM in MCF-7 breast cancer cells (ER+/PR+) and 74 nM in triple-negative MDA-MB-231 breast cancer cells. The compounds demonstrated significant G₂/M phase cell cycle arrest and induction of apoptosis in the MCF-7 cell line, inhibited tubulin polymerisation, and were selective for cancer cells when evaluated in non-tumorigenic MCF-10A breast cells. The immunofluorescence staining of MCF-7 cells confirmed that the compounds targeted tubulin and induced multinucleation, which is a recognised sign of mitotic catastrophe. Computational docking studies of compounds 19e, 21l, and 24 in the colchicine binding site of tubulin indicated potential binding conformations for the compounds. Compounds 19e and 21l were also shown to selectively inhibit aromatase. These compounds are promising candidates for development as antiproliferative, aromatase inhibitory, and microtubule-disrupting agents for breast cancer. Full article

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

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12 pages, 766 KiB

Open AccessReview

Non-Coding RNAs: The “Dark Side Matter” of the CLL Universe

by Marcello Francesco Lingua, Giovanna Carrà, Beatrice Maffeo and Alessandro Morotti

Pharmaceuticals 2021, 14(2), 168; https://doi.org/10.3390/ph14020168 - 21 Feb 2021

Cited by 5 | Viewed by 2246

Abstract

For many years in the field of onco-hematology much attention has been given to mutations in protein-coding genes or to genetic alterations, including large chromosomal losses or rearrangements. Despite this, biological and clinical needs in this sector remain unmet. Therefore, it is not [...] Read more.

For many years in the field of onco-hematology much attention has been given to mutations in protein-coding genes or to genetic alterations, including large chromosomal losses or rearrangements. Despite this, biological and clinical needs in this sector remain unmet. Therefore, it is not surprising that recent studies have shifted from coded to non-coded matter. The discovery of non-coding RNAs (ncRNAs) has influenced several aspects related to the treatment of cancer. In particular, in chronic lymphocytic leukemia (CLL) the knowledge of ncRNAs and their contextualization have led to the identification of new biomarkers used to follow the course of the disease, to the anticipation of mechanisms that support resistance and relapse, and to the selection of novel targeted treatment regimens. In this review, we will summarize the main ncRNAs discovered in CLL and the molecular mechanisms by which they are affected and how they influence the development and the progression of the disease. Full article

(This article belongs to the Special Issue Non-coding RNA in Hematological Cancers)

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40 pages, 8763 KiB

Open AccessReview

Advances in Development of Radiometal Labeled Amino Acid-Based Compounds for Cancer Imaging and Diagnostics

by Mária Bodnár Mikulová and Peter Mikuš

Pharmaceuticals 2021, 14(2), 167; https://doi.org/10.3390/ph14020167 - 21 Feb 2021

Cited by 15 | Viewed by 3433

Abstract

Radiolabeled biomolecules targeted at tumor-specific enzymes, receptors, and transporters in cancer cells represent an intensively investigated and promising class of molecular tools for the cancer diagnosis and therapy. High specificity of such biomolecules is a prerequisite for the treatment with a lower burden [...] Read more.

Radiolabeled biomolecules targeted at tumor-specific enzymes, receptors, and transporters in cancer cells represent an intensively investigated and promising class of molecular tools for the cancer diagnosis and therapy. High specificity of such biomolecules is a prerequisite for the treatment with a lower burden to normal cells and for the effective and targeted imaging and diagnosis. Undoubtedly, early detection is a key factor in efficient dealing with many severe tumor types. This review provides an overview and critical evaluation of novel approaches in the designing of target-specific probes labeled with metal radionuclides for the diagnosis of most common death-causing cancers, published mainly within the last three years. Advances are discussed such traditional peptide radiolabeling approaches, and click and nanoparticle chemistry. The progress of radiolabeled peptide based ligands as potential radiopharmaceuticals is illustrated via novel structure and application studies, showing how the molecular modifications reflect their binding selectivity to significant onco-receptors, toxicity, and, by that, practical utilization. The most impressive outputs in categories of newly developed structures, as well as imaging and diagnosis approaches, and the most intensively studied oncological diseases in this context, are emphasized in order to show future perspectives of radiometal labeled amino acid-based compounds in nuclear medicine. Full article

(This article belongs to the Special Issue Metal-Based Drugs: Updates and Perspectives)

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17 pages, 3684 KiB

Open AccessArticle

Losartan Improves Memory, Neurogenesis and Cell Motility in Transgenic Alzheimer’s Mice

by Henning Johannes Drews, Roman Klein, Ali Lourhmati, Marine Buadze, Elke Schaeffeler, Thomas Lang, Torgom Seferyan, Leah R. Hanson, William H. Frey II, Tom C.G.M. de Vries, Inge A.E.W. Thijssen-van Loosdregt, Christoph H. Gleiter, Matthias Schwab and Lusine Danielyan

Pharmaceuticals 2021, 14(2), 166; https://doi.org/10.3390/ph14020166 - 20 Feb 2021

Cited by 21 | Viewed by 4193

Abstract

Angiotensin receptor blockers (ARBs) have demonstrated multiple neuroprotective benefits in Alzheimer’s disease (AD) models. However, their beneficial effects on memory deficits, cholinergic activity, neurogenesis and Amyloid beta (Aβ) clearance reveal significant interstudy variability. The delivery route can impact not only delivery but also [...] Read more.

Angiotensin receptor blockers (ARBs) have demonstrated multiple neuroprotective benefits in Alzheimer’s disease (AD) models. However, their beneficial effects on memory deficits, cholinergic activity, neurogenesis and Amyloid beta (Aβ) clearance reveal significant interstudy variability. The delivery route can impact not only delivery but also targeting and therapeutic efficacy of ARBs. Our previous findings on the beneficial effects of intranasally delivered losartan in the APP/PS1 model of AD prompted us to explore the influence of the delivery route by employing here the systemic administration of losartan. Consistent with our previous results with intranasal losartan, repeated intraperitoneal administration (10 mg/kg) resulted in a remarkable decrease in Aβ plaques and soluble Aβ42, as well as inflammatory cytokines (IL-2, IL-6 and TNFα). The Aβ reduction can be ascribed to its facilitated degradation by neprilysin and diminished generation by BACE1. Losartan increased neurogenesis in vivo and in vitro and improved migratory properties of astrocytes isolated from adult transgenic AD mice. In summary, this data together with our previous results suggest therapeutic features of losartan which are independent of delivery route. The improvement of cell motility of Aβ-affected astrocytes by losartan deserves further in vivo investigation, which may lead to new strategies for AD treatment. Full article

(This article belongs to the Special Issue New Drugs and Biologics For Treatment of Central Nervous Dysfunction)

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Figure 1
Blood pressure (BP) and cognitive performance of losartan and vehicle treated APP/PS1 mice. Seven-month-old APP/PS1 mice (n = 9 in losartan and n = 8 in control group) were assessed for blood pressure and spatial working memory in losartan and vehicle treated (Ctrl.) groups. (a,b) systolic and diastolic blood pressure measured prior (baseline) and during drug (vehicle treatment (test blocks 1–3); (c) Latency to reach the goal arm and (d) number of incorrect choices in forced choice alternation T-maze assessed at the last 5 days of losartan or vehicle treatment. (c,d) Two-way ANOVA with Holm Sidak’s multiple comparisons test p < 0.05 (*), p < 0.01 (**) comparison between losartan and Ctrl. group; (##) comparison between day 1 and day 5 within the losartan-treated group. Full article ">Figure 2
Cerebral content of inflammatory cytokines and soluble beta Amyloid peptide 42 (Aβ42) in losartan and vehicle treated APP/PS1 mice. Brain homogenates of APP/PS1 mice (n = 5) were assessed for the levels of inflammatory cytokines. IL-2 (a), IL-6 (b), and TNFα (c) in losartan and vehicle treated (Ctrl.) groups. (d) Brain content of soluble Aβ42 in losartan and vehicle treated APP/PS1 mice. t-test, ** p < 0.01, *** p < 0.0005, **** p < 0.0001. Full article ">Figure 3
Losartan-mediated changes in neurogenesis and neuroprotection markers in APP/PS1 mice. (a,b) Expression of nestin (green) in the hilus and granular cell layer of the denate gyrus in brain sections of losartan vs. vehicle (Ctrl.) treated APP/PS1 mice. Cell nuclei are stained with 4´,6 diamidino-2-phenylindole (DAPI, blue), scale bar shown in (b) corresponds to the images shown in (a) and (b). (c) Cerebral content of Neuronal Differentiation 1 protein (NeuroD1), SRY-related HMG-box 2 (Sox2) and erythropoietin (EPO) assessed in brain homogenates of APP/PS1 mice (n = 4) by Western Blot. (d,e) Enzymatic activity (n = 8) and expression level of glutamine synthetase (GS) in brain homogenates of losartan and vehicle (Ctrl.) treated APP/PS1 mice (n = 4). Statistical analysis in (d): t-test, * p < 0.05. Full article ">Figure 4
Expression of factors regulating cholinergic activity, Aβ clearance and microglial function in losartan-treated APP/PS1 mice. (a) qPCR of β secretase 1 (BACE1) mRNA (n = 7 in control and n = 9 in losartan treated group); (b) Western blot of neprilysin, Acetylcholine esterase (AChE), Choline acetyltransferase (ChAT) and ionized calcium-binding adapter molecule 1 (Iba1), n = 4; (c–e) qPCR of AChE, ChAT and Iba1 mRNA (n = 7 in control and n = 9 in losartan treated group). Statistical analysis in (a) and (c–e): t-test, * p < 0.05, *** p < 0.0005. Full article ">Figure 5
Losartan-mediated changes in neurogenesis and neuroprotection markers in the hippocampus of APP/PS1 mice. (a,b) Immunofluorescent analysis of ChAT (green) and glial fibrillary acidic protein (GFAP) (red) in losartan vs. vehicle (Ctrl.) treated APP/PS1 mice. (c,d) Expression of AChE (red) and GFAP (green) in losartan/vehicle treated APP/PS1 mice. (e,f) Aβ plaques (red) and GFAP staining of losartan and vehicle (Ctrl.) treated APP/PS1 mouse brain sections. Cell nuclei are stained with DAPI (blue), scale bar in (b) corresponds to the images shown in (a) and (b), scale bar in (f) corresponds to (c–f). (a–f) Representative images out of 10 sections per brain taken from n = 3 mice analyzed per treatment group. Full article ">Figure 6
Losartan-induced neurogenesis and ACh production in wild type (WT) astroglial primary cultures (APC). (a,b) Immunofluorescent analysis of GFAP (green) and ACh (red) in losartan vs. vehicle (Ctrl.) treated WT APC. (c,d) Expression of β tubulin III (red) and GFAP (green) in losartan/vehicle treated WT APC. Arrow in (d) indicates young β tubulin III positive neuron in losartan treated culture. Cell nuclei are stained with DAPI (blue), scale bar in (d) corresponds to the images shown in (a–d). (a–d) Representative images out of n = 4 cover slips stained with ACh/GFAP or β tubulin III/GFAP in each treatment condition. Full article ">Figure 7
In vitro effects of losartan on triple transgenic Alzheimer’s disease (AD) mice (3xTg-AD) APC. (a,b) Western Blot of ChAT and BACE1 in losartan vs. vehicle (Ctrl.) treated 3xTg-APC. (b,c) Expression of neprilysin (green) in losartan (3xTg losartan) and vehicle (3xTg Ctrl.) treated 3xTg-APC. Cell nuclei are stained with DAPI (blue), scale bar in (d) corresponds to the images shown in (b,c). (d) Quantification of neprilysin positive cells in 3xTg-APC (n = 5). (e) Velocity of migrated cells in the control (ctrl.) and losartan treated 3xTg-APC. From the cells shown in videos in <a href="#app1-pharmaceuticals-14-00166" class="html-app">Supplementary Figure S1</a>, we identified and analyzed 32 cells per treatment group (n = 32) which were trackable for at least 12h; (f) accumulated distance calculated for cells described in (e); Statistical analysis in (d–f): t-test, * p < 0.05, *** p < 0.0005. Full article ">Figure 8
Schematic illustration of processes influenced by losartan in an AD-like pathology in vivo and in vitro. (a) In vivo effects of losartan in a transgenic model of AD (APP/PS1 mouse). Intraperitoneal administration of losartan led to a decrease in: Acetylcholine esterase (AchE), TNFα, IL-6, IL-2, β secretase 1 (BACE1), and beta amyloid (Aβ), and to an increase in Choline acetyltransferase (ChAT), glutamine synthetase (GS), neprilysin, SRY-related HMG-box 2 (Sox-2), Neuronal Differentiation 1 protein (NeuroD1) and erythropoietin (EPO). Blue solid arrows connect the processes (neurogenesis, neuroinflammation, cholinergic activity and Aβ/Glutamate (Glu)-toxicity) with their respective markers. Dashed blue arrows show the effect of EPO on GS, TNFα, IL-6 and neurogenesis evidenced by the literature indicated in white circles; (b) in vitro effects of losartan on the astrocytes and neural precursors. Losartan induced the differentiation of neurons from neural precursors, increased the Acetylcholine (ACh) in neurons, decreased the expression of BACE1 and upregulated ChAT and neprilysin in astrocytes. Losartan improved the motility and migration of astrocytes. This schematic drawing was created using art elements from Servier Medical Art Commons Attribution 3.0 Unported License. Servier Medical Art by Servier is licensed under a Creative Commons Attribution 3.0 Unported License. Full article ">

11 pages, 796 KiB

Open AccessArticle

Α Multicenter Retrospective Study Evaluating Brivaracetam in the Treatment of Epilepsies in Clinical Practice

by Maria Stefanatou, Eirini Vasileiadou Kapetanou, Vasilios K. Kimiskidis, Vasileios Papaliagkas, Panagiotis Polychronopoulos, Sofia Markoula, Kleoniki Charisiou, Dimitrios Kazis, Anastasia Verentzioti, Panayiotis Patrikelis, Athanasia Alexoudi and Stylianos Gatzonis

Pharmaceuticals 2021, 14(2), 165; https://doi.org/10.3390/ph14020165 - 19 Feb 2021

Cited by 4 | Viewed by 3278

Abstract

Brivaracetam (BRV) is the latest approved antiepileptic drug. The aim of the study was to evaluate the efficacy and tolerability of BRV in everyday clinical practice. In this retrospective, observational, multicenter study, data from epilepsy patients receiving BRV from January 2018 to July [...] Read more.

Brivaracetam (BRV) is the latest approved antiepileptic drug. The aim of the study was to evaluate the efficacy and tolerability of BRV in everyday clinical practice. In this retrospective, observational, multicenter study, data from epilepsy patients receiving BRV from January 2018 to July 2019 were analyzed. Patients with age ≥16 suffering from any type of epilepsy and having at least one follow up encounter after dose titration were included. 156 consecutive patients were included in the study. The mean age was 40 (16–84 years) and the mean duration of epilepsy was 21 years. Of the 156 patients, 81% were diagnosed with focal-onset seizures, 16% with generalized seizures, while 3% suffered from unclassified seizures. Nine patients received BRV as monotherapy as a switching therapy. At the first follow up visit, seizure cessation was achieved in 56 (36%) patients and the rate of ≥50% responders was 36%. Twenty four patients (15%) remained unchanged; six patients (4%) were recorded with increased seizure frequency, while the remaining 9% had a response of less than 50%. Twenty-six patients (17%) showed clinically significant adverse events, but none were life threatening. Brivaracetam seems to be an effective, easy to use and safe antiepileptic drug in the clinical setting. Full article

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

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18 pages, 2859 KiB

Open AccessArticle

In Vitro Characterization of Inhalable Cationic Hybrid Nanoparticles as Potential Vaccine Carriers

by Iman M. Alfagih, Kan Kaneko, Nitesh K. Kunda, Fars Alanazi, Sarah R. Dennison, Hesham M. Tawfeek and Imran Y. Saleem

Pharmaceuticals 2021, 14(2), 164; https://doi.org/10.3390/ph14020164 - 18 Feb 2021

Cited by 12 | Viewed by 3496

Abstract

In this study, PGA-co-PDL nanoparticles (NPs) encapsulating model antigen, bovine serum albumin (BSA), were prepared via double emulsion solvent evaporation. In addition, chitosan hydrochloride (CHL) was incorporated into the external phase of the emulsion solvent method, which resulted in surface adsorption onto the [...] Read more.

In this study, PGA-co-PDL nanoparticles (NPs) encapsulating model antigen, bovine serum albumin (BSA), were prepared via double emulsion solvent evaporation. In addition, chitosan hydrochloride (CHL) was incorporated into the external phase of the emulsion solvent method, which resulted in surface adsorption onto the NPs to form hybrid cationic CHL NPs. The BSA encapsulated CHL NPs were encompassed into nanocomposite microcarriers (NCMPs) composed of l-leucine to produce CHL NPs/NCMPs via spray drying. The CHL NPs/NCMPs were investigated for in vitro aerosolization, release study, cell viability and uptake, and stability of protein structure. Hybrid cationic CHL NPs (CHL: 10 mg/mL) of particle size (480.2 ± 32.2 nm), charge (+14.2 ± 0.72 mV), and BSA loading (7.28 ± 1.3 µg/mg) were produced. The adsorption pattern was determined to follow the Freundlich model. Aerosolization of CHL NPs/NCMPs indicated fine particle fraction (FPF: 46.79 ± 11.21%) and mass median aerodynamic diameter (MMAD: 1.49 ± 0.29 µm). The BSA α-helical structure was maintained, after release from the CHL NPs/NCMPs, as indicated by circular dichroism. Furthermore, dendritic cells (DCs) and A549 cells showed good viability (≥70% at 2.5 mg/mL after 4–24 h exposure, respectively). Confocal microscopy and flow cytometry data showed hybrid cationic CHL NPs were successfully taken up by DCs within 1 h of incubation. The upregulation of CD40, CD86, and MHC-II cell surface markers indicated that the DCs were successfully activated by the hybrid cationic CHL NPs. These results suggest that the CHL NPs/NCMPs technology platform could potentially be used for the delivery of proteins to the lungs for immunostimulatory applications such as vaccines. Full article

(This article belongs to the Special Issue Nano Drug Carriers 2021)

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Graphical abstract

Graphical abstract
Full article ">Figure 1
The relationship of CHL concentration to the amount absorbed on PGA-co-PDL NPs (A). Zeta potential and particle size of hybrid cationic CHL NPs (B) (n = 3, p < 0.05 is indicated by *). Full article ">Figure 2
Linear description of CHL adsorption onto PGA-co-PDL NPs. Ce is the residual CHL concentrations in the suspension at equilibrium (mg/mL) and q is the amount of adsorbed CHL per unit weight of NPs. All models (A) Langmuir, (B) BET, (C) Freundlich (D) Halsey were fit utilizing concentrations of 2–20 mg/mL; however, the Langmuir model was also fit to low concentrations (0–8 mg/mL) (black) (n = 3). Full article ">Figure 3
Scanning electron microscopy images of cationic CHL NPs/NCMPs: (A) 1 µm scale and (B) 2 µm scale. Full article ">Figure 4
In vitro release of BSA from cationic CHL NPs/NCMPs in PBS buffer at 37 °C (n = 3). Full article ">Figure 5
SDS-PAGE to evaluate the BSA stability released from cationic CHL NPs/NCMPs (A). Lanes represent, molecular weight (MW) standard markers, BSA (MW 66,000) (1), BSA standard (2), BSA released from hybrid cationic CHL NPs (3), and BSA released from cationic CHL NPs/NCMPs (4). Difference in band intensity was due to different loading. (The CD spectra of BSA released from cationic CHL NPs/NCMPs (grey) and BSA standard (black) (B) (n = 3). Full article ">Figure 6
MTT assay to determine cell viability of A549 cells after 24 h of exposure to hybrid cationic CHL NPs and cationic CHL NPs/NCMPs, and DCs after 4 h of exposure to hybrid cationic CHL NPs (n = 3). Full article ">Figure 7
Confocal microscopy images of hybrid cationic CHL NPs uptake by DCs after 1 h incubation. DCs incubated without hybrid cationic CHL NPs at 20× (A), DCs incubated with hybrid cationic CHL NPs at 63× (B). Red channel for WGA TR: cell membrane, blue channel for DAPI: nucleus, and green channel for FITC-BSA: hybrid cationic CHL NPs incorporating FITC-BSA. Full article ">Figure 8
The association/uptake of FITC-BSA in cationic CHL NPs, by DCs. DCs were incubated with FITC-BSA incorporated CHL NPs for either 1 or 4 h, at either 4 or 37 °C (n = 4). Full article ">Figure 9
Upregulation of cell surface CD40 (A), CD86 (B), and MHC-II (C) on DCs, and cell viability according to 7AAD staining (D), after incubation with BSA incorporated hybrid cationic CHL NPs for 24 h (n = 6). The indicated groups exhibited significantly greater MFI (p < 0.05) compared to negative control. (1), BSA (2), CHL NPs (3), BSA CHL NPs (4), LPS (5). Full article ">

10 pages, 1632 KiB

Open AccessArticle

Development and Validation of an LC-MS/MS Method for Quantification of the Novel Antibacterial Candidate DA-7010 in Plasma and Application to a Preclinical Pharmacokinetic Study

by Mi Hye Kwon, Dae Young Lee and Hee Eun Kang

Pharmaceuticals 2021, 14(2), 163; https://doi.org/10.3390/ph14020163 - 18 Feb 2021

Cited by 2 | Viewed by 2683

Abstract

DA-7010 is a new candidate for an antibacterial agent that targets Gram-negative pathogens by acting as a leucyl-tRNA synthetase inhibitor. In this study, a simple and rapid liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed to determine DA-7010 levels in the plasma [...] Read more.

DA-7010 is a new candidate for an antibacterial agent that targets Gram-negative pathogens by acting as a leucyl-tRNA synthetase inhibitor. In this study, a simple and rapid liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed to determine DA-7010 levels in the plasma from mice, rats, and dogs. Plasma samples were mixed with methanol for protein precipitation. Chromatographic separation was carried out using a reversed-phase C₁₈ column (Agilent Poroshell 120, 50 × 3.0 mm, 2.7 μm). An isocratic elution of the mobile phase consisting of 5 mM formic acid in water and acetonitrile at a ratio of 84:16 (v/v) was applied at a flow rate of 0.3 mL/min. The total chromatographic run time was 3.5 min. Multiple reaction monitoring (MRM) mode was used for mass spectrometric detection using an Agilent 6460 triple quadrupole coupled with an electrospray ionization (ESI) source operated in positive-ion mode. The MRM transitions analyzed were m/z 220.1→162.1 for DA-7010 and m/z 206.1→170.1 for the internal standard (structural analogue of DA-7010). Calibration curves were constructed in the range of 10–10,000 ng/mL. The intra- and interday precision and accuracy were within 11.3%, excluding those for the lower limit of quantification (LLOQ) samples, which were within 17.1%. The developed LC-MS/MS method was successfully validated and applied in preclinical pharmacokinetic studies of DA-7010 in mice, rats, and dogs following single oral administrations. The oral absorption of DA-7010 was rapid, and the systemic exposure was approximately four times higher in the dogs than in the mice or rats. Full article

(This article belongs to the Special Issue Analytical Techniques in the Pharmaceutical Sciences)

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13 pages, 1929 KiB

Open AccessArticle

Limited Sampling Strategy for Determination of Ibrutinib Plasma Exposure: Joint Analyses with Metabolite Data

by Félicien Le Louedec, Fanny Gallais, Fabienne Thomas, Mélanie White-Koning, Ben Allal, Caroline Protin, Loïc Ysebaert, Étienne Chatelut and Florent Puisset

Pharmaceuticals 2021, 14(2), 162; https://doi.org/10.3390/ph14020162 - 18 Feb 2021

Cited by 5 | Viewed by 3814

Abstract

Therapeutic drug monitoring of ibrutinib is based on the area under the curve of concentration vs. time (AUC_IBRU) instead of trough concentration (C_min,ss) because of a limited accumulation in plasma. Our objective was to identify a limited sampling strategy [...] Read more.

Therapeutic drug monitoring of ibrutinib is based on the area under the curve of concentration vs. time (AUC_IBRU) instead of trough concentration (C_min,ss) because of a limited accumulation in plasma. Our objective was to identify a limited sampling strategy (LSS) to estimate AUC_IBRU associated with Bayesian estimation. The actual AUC_IBRU of 85 patients was determined by the Bayesian analysis of the full pharmacokinetic profile of ibrutinib concentrations (pre-dose T0 and 0.5, 1, 2, 4 and 6 h post-dose) and experimental AUC_IBRU were derived considering combinations of one to four sampling times. The T0–1–2–4 design was the most accurate LSS (root-mean-square error RMSE = 11.0%), and three-point strategies removing the 1 h or 2 h points (RMSE = 22.7% and 14.5%, respectively) also showed good accuracy. The correlation between the actual AUC_IBRU and C_min,ss was poor (r² = 0.25). The joint analysis of dihydrodiol-ibrutinib metabolite concentrations did not improve the predictive performance of AUC_IBRU. These results were confirmed in a prospective validation cohort (n = 27 patients). At least three samples, within the pre-dose and 4 h post-dose period, are necessary to estimate ibrutinib exposure accurately. Full article

(This article belongs to the Special Issue Precision Medicine in Oncology: Controlling the Pharmacokinetics Variability)

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16 pages, 1833 KiB

Open AccessReview

Bacteriophages as Therapeutic and Diagnostic Vehicles in Cancer

by Valentina Foglizzo and Serena Marchiò

Pharmaceuticals 2021, 14(2), 161; https://doi.org/10.3390/ph14020161 - 17 Feb 2021

Cited by 36 | Viewed by 7214

Abstract

Evolution of nanomedicine is the re-design of synthetic and biological carriers to implement novel theranostic platforms. In recent years, bacteriophage research favors this process, which has opened up new roads in drug and gene delivery studies. By displaying antibodies, peptides, or proteins on [...] Read more.

Evolution of nanomedicine is the re-design of synthetic and biological carriers to implement novel theranostic platforms. In recent years, bacteriophage research favors this process, which has opened up new roads in drug and gene delivery studies. By displaying antibodies, peptides, or proteins on the surface of different bacteriophages through the phage display technique, it is now possible to unravel specific molecular determinants of both cancer cells and tumor-associated microenvironmental molecules. Downstream applications are manifold, with peptides being employed most of the times to functionalize drug carriers and improve their therapeutic index. Bacteriophages themselves were proven, in this scenario, to be good carriers for imaging molecules and therapeutics as well. Moreover, manipulation of their genetic material to stably vehiculate suicide genes within cancer cells substantially changed perspectives in gene therapy. In this review, we provide examples of how amenable phages can be used as anticancer agents, especially because their systemic administration is possible. We also provide some insights into how their immunogenic profile can be modulated and exploited in immuno-oncology for vaccine production. Full article

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

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17 pages, 2631 KiB

Open AccessArticle

Design and Mechanism of Action of a New Prototype of Combi-Molecule “Programed” to Release Bioactive Species at a pH Range Akin to That of the Tumor Microenvironment

by Anne-Laure Larroque-Lombard, Etienne Chatelut, Jean-Pierre Delord, Diane-Charlotte Imbs, Philippe Rochaix, Bertrand Jean-Claude and Ben Allal

Pharmaceuticals 2021, 14(2), 160; https://doi.org/10.3390/ph14020160 - 16 Feb 2021

Cited by 3 | Viewed by 3863

Abstract

The clinical use of cytotoxic agents is plagued by systemic toxicity. We report a novel approach that seeks to design a “combi-molecule” to behave as an alkylating agent on its own and to undergo acid-catalyzed conversion to two bioactive species at a pH [...] Read more.

The clinical use of cytotoxic agents is plagued by systemic toxicity. We report a novel approach that seeks to design a “combi-molecule” to behave as an alkylating agent on its own and to undergo acid-catalyzed conversion to two bioactive species at a pH range akin to that of a tumor microenvironment: an AL530 prototype was synthesized and we studied its ability to release a chlorambucil analogue (CBL-A) plus a potent mitogen-activated protein/extracellular signal-regulated kinase kinase (MEK) inhibitor (PD98059) at different pHs in buffered solutions, plasma and tumors. Its potency was compared in vitro with CBL+PD98059 (SRB assay) and in vivo in a xenograft model. Its target modulation was studied by western blotting and immunohistochemistry. AL530 released PD98059+CBL-A at mild acidic pH and in vitro was fivefold more potent than CBL and three-to-fivefold more potent than CBL+PD98059. In vivo it released high levels of PD98059 in tumors with a tumor/plasma ratio of five. It induced γ-H2AX phosphorylation and blocked pErk1,2, indirectly indicating its ability to damage DNA and modulate MEK. It induced significant tumor delay and less toxicity at unachievable doses for CBL and CBL+PD98059. We demonstrated the feasibility of a pH-labile combi-molecule capable of delivering high MEK inhibitor concentration in tumors, damaging DNA therein, and inducing tumor growth delay. Full article

(This article belongs to the Special Issue Precision Medicine in Oncology: Controlling the Pharmacokinetics Variability)

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Figure 1
Multiple cell signaling pathways leading to MEK activation. GF: Growth Factor, RTK: Receptor Tyrosine Kinase. Full article ">Figure 2
Influence of pH and temperature on AL530 hydrolysis. Graphs (A) and (B) represent the formation rate (constant k) of PD98059 at ■—37 °C, ▲—40 °C and ▼—42 °C versus the pH ranging from 5.5 to 7. The overlaid chromatograms (C) represent the time course effects (from 24 to 96 h) of AL530 hydrolysis versus the formation of PD98059 at 37 °C at pH = 7. Full article ">Figure 3
Study of Erk1,2 phosphorylation inhibition in CAL33 cells daily treated with AL530 (25 µM) for 24 to 48 h. At the indicated times in the figure, cells were harvested, lysed (RIPA buffer), an equal amount of protein was separated (12.5% SDS PAGE gel), blotted (PVDF membrane) and coated with antibody of interest. Full article ">Figure 4
(A) In vivo hydrolysis of AL530 in 4T1 mouse mammary tumors (top panel) and human head and neck tumors generated from the CAL33 human xenograft in nude mice (bottom panel). Each chromatogram represents the analysis of plasma or tumor extract from a mouse treated with 100 mg/kg of AL530 (i.p.) after 3 h (4T1 model) and 50 mg/kg (i.p.) after 1 h (CAL33 model). Following mice sacrifice, plasma and tissue were collected and stored at −80 °C until the day of analysis by HPLC as described in the Materials and Methods section. Consistently higher levels of PD98059 were released in tumor tissues than in plasma. (B) Proposed model for the absorption and distribution of AL530 and the localization of its hydrolytic conversion to PD98059 + CBL-A. Full article ">Figure 5
Immunohistochemistry analysis of the effect of AL530 on tumor tissue. Thick sections of 4 µm formalin fixed and paraffin embedded in tumors from CAL33 cell xenografts in nude mice treated with vehicle or 50 mg/kg of AL530 i.p. for 13 days, were stained with antibody against γ-H2AX (A) and pErk1,2 (B) (magnification, x20). Full article ">Figure 6
Effect of vehicle, AL530, Chlorambucil (CBL), PD98059 (MEK inhibitor) or equimolar combination CBL + PD98059 treatment on tumors growth (A,C) and weight (B) properties of CAL33 head and neck cancer cell xenograft. Nude mice (n = 7 per group) bearing subcutaneous 200 mm3 (at the start of the experiment) bilateral CAL33 tumors xenograft received either saline (orange square, A1), 25 mg/kg (A1) or 50 mg/kg (C) of AL530 (green triangle), or 11 mg/kg of CBL (A2, blue diamond), or 9.7 mg/kg of PD98059 (data not shown) or an equimolar combination of CBL + PD98059 (1:1; A3, orange circle) daily by i.p. route/q5d throughout the experiment. Data are expressed as mean ± SEM. ** p < 0.0001 and * p < 0.01, Mann–Whitney posthoc test. Full article ">Scheme 1
2-Nitroaryl amides proposed as bioreducible prodrugs of anilines based on the intramolecular cyclization of the 2-aminoaryl amides resulting from the reduction of the 2-nitro group. Full article ">Scheme 2
Proposed mechanism for the pH dependence of the degradation of the lead combi-molecule AL530. Full article ">Scheme 3
(i) 8S, 1 h and THF/CH2Cl2 1/1, Et3N, 0 °C to 50 °C, 3 days, 44%, (ii) LiOH 2N, MeOH, rt, 18 h, 97%, (iii) CDI, aniline mustard (compound 12S), THF/ACN 1/1, 0 °C and rt, 18 h, 47%, (iv) Pd/C 10%, MeOH, H2, rt, 3 h, 65%. Full article ">

9 pages, 585 KiB

Open AccessCase Report

Tumor Type Agnostic Therapy Carrying BRAF Mutation: Case Reports and Review of Literature

by Ottavia Bernocchi, Marianna Sirico, Silvia Paola Corona, Carla Strina, Manuela Milani, Maria Rosa Cappelletti, Giuseppina Ferrero, Nicoletta Ziglioli, Valeria Cervoni, Andrea Macchiavelli, Giandomenico Roviello and Daniele Generali

Pharmaceuticals 2021, 14(2), 159; https://doi.org/10.3390/ph14020159 - 16 Feb 2021

Cited by 6 | Viewed by 3603

Abstract

Background: Precision medicine is based on molecular and genotypic patient characterization to define specific target treatment. BRAF mutation is an oncogenic driver, and the Cancer Genome Atlas has identified BRAF mutations in different cancer types. Tumor type agnostic therapy is based on targeting [...] Read more.

Background: Precision medicine is based on molecular and genotypic patient characterization to define specific target treatment. BRAF mutation is an oncogenic driver, and the Cancer Genome Atlas has identified BRAF mutations in different cancer types. Tumor type agnostic therapy is based on targeting genomic alterations, regardless of tumor origin. In this context, novel therapeutic agents including BRAF and MEK inhibitors based on the molecular landscape in solid tumors have been investigated. Case presentation, Case 1: The first case is chemotherapy-refractory, BRAF V600E mutated intrahepaticcholangiocarcinoma treated with vemurafenib and cobimetinib as third line therapy. In this setting the dual BRAF and MEK inhibition resulted in improved progression-free survival and quality of life; Case 2: The second case shows aBRAF G466A mutated Bellini duct carcinoma (BDC), treated with dabrafenib and trametinib in second line therapy. The disease remained under control for 11 months after the first relapse. Discussion: In the literature there is strong evidence that melanoma, colorectal cancer, non small cell lung cancer and anaplastic thyroid cancer with BRAF mutations are good targets for BRAF/MEK pathway inhibitors. The VE-BASKET and ROAR basket trials explored the efficacy of vemurafenib and the combination of dabrafenib/trametinib, respectively, in BRAF V600 mutation-positive cancers other than melanoma, papillary thyroid cancer, colorectal cancer and non small cell lung cancer. Within the concept of tumor type agnostic therapy, we decided to treat our BRAF-mutated tumors with the association of BRAF and MEK inhibitors. Conclusions: Our results confirm the emerging importance of molecular tumor profiling for the successful management of cancer, and the potential of BRAF-targeted therapy in the treatment of rare solid tumors with poor prognosis and no clinical benefit from systemic therapies with. Full article

(This article belongs to the Special Issue Cancer Translational Biomarkers and Targeted Therapies)

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13 pages, 1210 KiB

Open AccessArticle

Preparation and Bioevaluation of Novel ^99mTc-Labeled Complexes with a 2-Nitroimidazole HYNIC Derivative for Imaging Tumor Hypoxia

by Qing Ruan, Qianqian Gan, Xuran Zhang, Si’an Fang and Junbo Zhang

Pharmaceuticals 2021, 14(2), 158; https://doi.org/10.3390/ph14020158 - 15 Feb 2021

Cited by 6 | Viewed by 3189

Abstract

To develop novel ^99mTc-labeled single-photon emission computed tomography (SPECT) radiotracers for imaging hypoxia, a novel HYNICNM ligand (6-hydrazinonicotinamide (HYNIC) 2-nitroimidazole derivative) was designed and synthesized. It was radiolabeled with technetium-99m using tricine/trisodium triphenylphosphine-3,3′,3′′-trisulfonate (TPPTS), tricine/sodium triphenylphosphine-3-monosulfonate (TPPMS) and tricine as co-ligands to [...] Read more.

To develop novel ^99mTc-labeled single-photon emission computed tomography (SPECT) radiotracers for imaging hypoxia, a novel HYNICNM ligand (6-hydrazinonicotinamide (HYNIC) 2-nitroimidazole derivative) was designed and synthesized. It was radiolabeled with technetium-99m using tricine/trisodium triphenylphosphine-3,3′,3′′-trisulfonate (TPPTS), tricine/sodium triphenylphosphine-3-monosulfonate (TPPMS) and tricine as co-ligands to obtain [^99mTc]Tc-tricine-TPPTS-HYNICNM, [^99mTc]Tc-tricine-TPPMS-HYNICNM, and [^99mTc]Tc-(tricine)₂-HYNICNM, respectively. The three technetium-99m complexes were radiolabeled in one step with a high yield (95%) and had good stability in saline and mouse serum. In vitro cellular uptake results showed that these complexes exhibited good hypoxic selectivity. The partition coefficient indicated that they were good hydrophilic complexes, and [^99mTc]Tc-tricine-TPPTS-HYNICNM displayed the highest hydrophilicity (−3.02 ± 0.08). The biodistribution in mice bearing S180 tumors showed that [^99mTc]Tc-tricine-TPPTS-HYNICNM exhibited higher tumor uptake (1.05 ± 0.27% IA/g); more rapid clearance from the liver, blood, muscle, and other non-target organs; and a higher tumor/non-target ratio, especially for the tumor/liver ratio (1.95), than [^99mTc]Tc-tricine-TPPMS-HYNICNM and [^99mTc]Tc-(tricine)₂-HYNICNM. The results of single-photon emission computed tomography (SPECT) imaging studies of [^99mTc]Tc-tricine-TPPTS-HYNICNM were in accordance with the biodistribution results, which suggested that [^99mTc]Tc-tricine-TPPTS-HYNICNM is a promising agent for imaging tumor hypoxia. Full article

(This article belongs to the Special Issue Forward Thinking towards Theranostic (Radio)ligands Targeting the Tumor Microenvironment (TME))

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28 pages, 6335 KiB

Open AccessReview

Status and Challenges of Plant-Anticancer Compounds in Cancer Treatment

by Paula Garcia-Oliveira, Paz Otero, Antia Gonzalez Pereira, Franklin Chamorro, Maria Carpena, Javier Echave, Maria Fraga-Corral, Jesus Simal-Gandara and Miguel Angel Prieto

Pharmaceuticals 2021, 14(2), 157; https://doi.org/10.3390/ph14020157 - 14 Feb 2021

Cited by 136 | Viewed by 12813

Abstract

Nowadays, cancer is one of the deadliest diseases in the world, which has been estimated to cause 9.9 million deaths in 2020. Conventional treatments for cancer commonly involve mono-chemotherapy or a combination of radiotherapy and mono-chemotherapy. However, the negative side effects of these [...] Read more.

Nowadays, cancer is one of the deadliest diseases in the world, which has been estimated to cause 9.9 million deaths in 2020. Conventional treatments for cancer commonly involve mono-chemotherapy or a combination of radiotherapy and mono-chemotherapy. However, the negative side effects of these approaches have been extensively reported and have prompted the search of new therapeutic drugs. In this context, scientific community started to look for innovative sources of anticancer compounds in natural sources, including traditional plants. Currently, numerous studies have evaluated the anticancer properties of natural compounds derived from plants, both in vitro and in vivo. In pre-clinical stages, some promising compounds could be mentioned, such as the sulforaphane or different phenolic compounds. On the other hand, some phytochemicals obtained positive results in clinical stages and were further approved for cancer treatment, such as vinca alkaloids or the paclitaxel. Nevertheless, these compounds are not exempt of limitations, such as low solubility, restricted effect on their own, negative side-effects, etc. This review aims to compile the information about the current phytochemicals used for cancer treatment and also promising candidates, main action mechanisms and also reported limitations. In this sense, some strategies to face the limitations have been considered, such as nano-based formulations to improve solubility or chemical modification to reduce toxicity. In conclusion, although more research is still necessary to develop more efficient and safe phytochemical drugs, more of these compounds might be used in future cancer therapies. Full article

(This article belongs to the Special Issue Anticancer Compounds in Medicinal Plants)

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8 pages, 1294 KiB

Open AccessArticle

Quetiapine-Induced Place Preference in Mice: Possible Dopaminergic Pathway

by Yusuf S. Althobaiti

Pharmaceuticals 2021, 14(2), 156; https://doi.org/10.3390/ph14020156 - 14 Feb 2021

Cited by 4 | Viewed by 2806

Abstract

Quetiapine, an atypical antipsychotic, is effective in the management of schizophrenia, depression, and anxiety. Although quetiapine overdosage and misuse have been reported, its abuse potential has not been investigated in animals. In this study, the abuse potential of quetiapine was assessed based on [...] Read more.

Quetiapine, an atypical antipsychotic, is effective in the management of schizophrenia, depression, and anxiety. Although quetiapine overdosage and misuse have been reported, its abuse potential has not been investigated in animals. In this study, the abuse potential of quetiapine was assessed based on the conditioned place preference (CPP) paradigm of drug addiction in a mouse model. First, mice received intraperitoneal injections of quetiapine (40, 80, or 120 mg/kg) every other day during the conditioning phase. In the second experiment, mice were pretreated with 0.03 mg/kg SKF-35866, a D1 receptor antagonist, before receiving saline or quetiapine (120 mg/kg) during the conditioning phase. No significant changes in time spent in the quetiapine-paired chamber were observed compared with time spent in the saline-paired chamber in mice treated with 40 or 80 mg/kg. In contrast, the preference to the quetiapine-paired chamber was significantly increased in mice treated with 120 mg/kg quetiapine, and this effect was blocked by SKF-35866 pretreatment. These results demonstrated, for the first time, the abuse potential of quetiapine in an animal model of drug addiction. Interestingly, this CPP-inducing effect was likely mediated by activating D1 receptors. Full article

(This article belongs to the Section Medicinal Chemistry)

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(A) Time spent in chamber 1 compared with chamber 2 during the pretest and post-test in the control group. (B) Time spent in the quetiapine-paired chamber compared with the saline-paired chamber during the pretest and post-test in the Quet-40 group (treated with quetiapine 40 mg/kg). Values are shown as means ± standard errors of the means. Full article ">Figure 2
(A) Time spent in the quetiapine-paired chamber compared with the saline-paired chamber during pretest and post-test in the Quet-80 group (treated with quetiapine; 80 mg/kg). (B) Time spent in the quetiapine-paired chamber compared with the saline-paired chamber during pretest and post-test in the Quet-120 group (treated with quetiapine; 120 mg/kg). Values are shown as means ± standard errors of the means. *p < 0.05 compared with the saline-paired chamber. Full article ">Figure 3
(A) Time spent in the D1 antagonist-paired chamber compared with the saline-paired chamber during pretest and post-test in the SKF-V group (treated with SKF followed by vehicle). (B) Time spent in the quetiapine-paired chamber compared with the saline-paired chamber during pretest and post-test in the SKF-Quet group (treated with SKF followed by quetiapine). Values are shown as means ± standard errors of the means. Full article ">Figure 4
Experimental schedule of the conditioned place preference (CPP) experiments. Full article ">

18 pages, 3040 KiB

Open AccessArticle

Synthesis, Characterization, and Biological Evaluation of New Derivatives Targeting MbtI as Antitubercular Agents

by Matteo Mori, Giovanni Stelitano, Laurent R. Chiarelli, Giulia Cazzaniga, Arianna Gelain, Daniela Barlocco, Elena Pini, Fiorella Meneghetti and Stefania Villa

Pharmaceuticals 2021, 14(2), 155; https://doi.org/10.3390/ph14020155 - 13 Feb 2021

Cited by 24 | Viewed by 4060

Abstract

Tuberculosis (TB) causes millions of deaths every year, ranking as one of the most dangerous infectious diseases worldwide. Because several pathogenic strains of Mycobacterium tuberculosis (Mtb) have developed resistance against most of the established anti-TB drugs, new therapeutic options are urgently needed. An [...] Read more.

Tuberculosis (TB) causes millions of deaths every year, ranking as one of the most dangerous infectious diseases worldwide. Because several pathogenic strains of Mycobacterium tuberculosis (Mtb) have developed resistance against most of the established anti-TB drugs, new therapeutic options are urgently needed. An attractive target for the development of new antitubercular agents is the salicylate synthase MbtI, an essential enzyme for the mycobacterial siderophore biochemical machinery, absent in human cells. A set of analogues of I and II, two of the most potent MbtI inhibitors identified to date, was synthesized, characterized, and tested to elucidate the structural requirements for achieving an efficient MbtI inhibition and a potent antitubercular activity with this class of compounds. The structure-activity relationships (SAR) here discussed evidenced the importance of the furan as part of the pharmacophore and led to the preparation of six new compounds (IV–IX), which gave us the opportunity to examine a hitherto unexplored position of the phenyl ring. Among them emerged 5-(3-cyano-5-(trifluoromethyl)phenyl)furan-2-carboxylic acid (IV), endowed with comparable inhibitory properties to the previous leads, but a better antitubercular activity, which is a key issue in MbtI inhibitor research. Therefore, compound IV offers promising prospects for future studies on the development of novel agents against mycobacterial infections. Full article

(This article belongs to the Special Issue Novel Antibacterial Agents)

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Reactions catalyzed by MbtI. Full article ">Figure 2
Chemical structure of the lead compounds I, II, and III. Full article ">Figure 3
Chemical structure of the heterocyclic cores tested in this study: 1 (thiophene), 2 (thiazole), 3 (oxazole), 4 (imidazole), 5 (1,3,4-oxadiazole), 6 (1,2,3-triazole). Full article ">Figure 4
Chemical structure of compounds IV–IX. Full article ">Figure 5
Biological characterization of IV. (A) IC50 determination of IV against MbtI activity. (B) Global reciprocal plot of data from MbtI steady-state kinetics analysis towards chorismic acid, in the presence of different concentrations of IV (50, 20, 10, 5, 1, and 0 μM). (C) MIC99 determination of IV against M. bovis BCG growth. Full article ">Scheme 1
Synthetic procedure for the preparation of 1a,b. Reagents and conditions: (a) MeOH, conc. H2SO4, reflux, overnight; (b) Pd(PPh3)2Cl2, 2 M Na2CO3, dry 1,4-dioxane, 90 °C, overnight, N2 atm; (c) 1. LiOH, THF-H2O 2:1, r.t., 2 h for 1a; 1 M NaOH, EtOH-THF 1:1, reflux, 5 h for 1b; 2. 1 M HCl, 0 °C. Full article ">Scheme 2
Synthetic procedure for the preparation of 2a,b. Reagents and conditions: (a) NBS, p-TsOH, DCM, overnight, r.t, N2 atm.; (b) 1. hexamine, DCM, 8 h, r.t.; 2. conc. HCl, EtOH, overnight, r.t.; (c) TEA, EtOAc, 3 h, reflux; (d) Lawesson’s reagent, 1,4-dioxane, 2 h, reflux; (e) NaOH, THF-H2O 1:1, 1.5 h, r.t. Full article ">Scheme 3
Synthetic procedure for the preparation of 3a,b. Reagents and conditions: (a) I2, DMSO, 3 h, 130 °C; (b) NaOH, THF-H2O 1:1, 1.5 h, r.t. Full article ">Scheme 4
Synthetic procedure for the preparation of 4a,b. Reagents and conditions: (a) SeO2, 1,4-dioxane/H2O, reflux, 7 h, N2 atm; (b) NH4OAc, CH3CN, H2O, r.t., 2 h; (c) 1. LiOH, THF-H2O 2:1, r.t., overnight for 4a; NaOH, THF-H2O 1:1, reflux, 6 h for 4b; 2. 3 M HCl, 0 °C. Full article ">Scheme 5
(A) Synthetic procedure for the preparation of 5a. Reagents and conditions: (a) dry MeOH, conc. H2SO4, reflux, 3 h, N2 atm; (b) NH2NH2∙H2O, MeOH, r.t., overnight; (c) TEA, DCM, r.t., 2 h; (d) TEA, DCM, TsCl, r.t., 2 h; (e) 1. NaOH, THF-H2O 1:1, r.t., 1 h; 2. Amberlite IR120, 0 °C. (B) Synthetic procedure for the preparation of 5b. Reagents and conditions: (a) EtOH, conc. H2SO4, reflux, overnight; (b) NH2NH2∙H2O, EtOH, reflux, overnight; (c) 86% PPA, 120-130 °C, 1.5 h; (d) 1. LiOH, THF-H2O 1:1, r.t., 1 h; 2. Amberlite IR120, 0 °C. Full article ">Scheme 6
Synthetic procedure for the preparation of 6a,b. Reagents and conditions: (a) 1. NaN3, Cu(OAc)2, MeOH, 55 °C, 1.5-4 h, N2 atm; 2. ethyl propiolate, (+)-sodium l-ascorbate, r.t., overnight-24 h; (b) 1. LiOH, THF-H2O 2:1, r.t., 1 h for 6a; NaOH, THF-H2O 1:1, reflux, 5 h for 6b; 2. 3 M HCl, 0 °C for 6a; 1 M HCl, 0 °C for 6b. Full article ">

12 pages, 304 KiB

Open AccessReview

The Treatment of Lung Involvement in Systemic Sclerosis

by Barbara Ruaro, Marco Confalonieri, Marco Matucci-Cerinic, Francesco Salton, Paola Confalonieri, Mario Santagiuliana, Gloria Maria Citton, Elisa Baratella and Cosimo Bruni

Pharmaceuticals 2021, 14(2), 154; https://doi.org/10.3390/ph14020154 - 13 Feb 2021

Cited by 17 | Viewed by 3394

Abstract

Systemic sclerosis (SSc) patients are often affected by interstitial lung disease (ILD) and, although there have been recent treatment advances, it remains the leading cause of death among SSc, with a 10-year mortality up to 40%. African Americans and subjects with diffuse cutaneous [...] Read more.

Systemic sclerosis (SSc) patients are often affected by interstitial lung disease (ILD) and, although there have been recent treatment advances, it remains the leading cause of death among SSc, with a 10-year mortality up to 40%. African Americans and subjects with diffuse cutaneous SSc or anti-topoisomerase 1 antibodies are most commonly affected. Currently, early ILD diagnosis can be made, and it is pivotal to improve the prognosis. The diagnostic mainstay test for SSc-ILD is high-resolution computed tomography for the morphology and pulmonary function tests for the functional aspects. Treatment planning and intensity are guided by the disease severity and risk of progression. Traditionally, therapy has depended on combinations of immunosuppressants, particularly cyclophosphamide and mycophenolate mofetil, which can be supplemented by targeted biological and antifibrotic therapies. Benefits have been observed in trials on hematopoietic autologous stem cell transplantation for patients with progressive SSc, whilst lung transplantation is reserved for refractory SSc-ILD cases. Herein, recent advances in SSc-ILD treatment will be explored. Full article

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

17 pages, 3619 KiB

Open AccessArticle

Pharyngeal Pumping and Tissue-Specific Transgenic P-Glycoprotein Expression Influence Macrocyclic Lactone Susceptibility in Caenorhabditis elegans

by Alexander P. Gerhard, Jürgen Krücken, Cedric Neveu, Claude L. Charvet, Abdallah Harmache and Georg von Samson-Himmelstjerna

Pharmaceuticals 2021, 14(2), 153; https://doi.org/10.3390/ph14020153 - 13 Feb 2021

Cited by 13 | Viewed by 3522

Abstract

Macrocyclic lactones (MLs) are widely used drugs to treat and prevent parasitic nematode infections. In many nematode species including a major pathogen of foals, Parascaris univalens, resistance against MLs is widespread, but the underlying resistance mechanisms and ML penetration routes into nematodes [...] Read more.

Macrocyclic lactones (MLs) are widely used drugs to treat and prevent parasitic nematode infections. In many nematode species including a major pathogen of foals, Parascaris univalens, resistance against MLs is widespread, but the underlying resistance mechanisms and ML penetration routes into nematodes remain unknown. Here, we examined how the P-glycoprotein efflux pumps, candidate genes for ML resistance, can modulate drug susceptibility and investigated the role of active drug ingestion for ML susceptibility in the model nematode Caenorhabditis elegans. Wildtype or transgenic worms, modified to overexpress P. univalens PGP-9 (Pun-PGP-9) at the intestine or epidermis, were incubated with ivermectin or moxidectin in the presence (bacteria or serotonin) or absence (no specific stimulus) of pharyngeal pumping (PP). Active drug ingestion by PP was identified as an important factor for ivermectin susceptibility, while moxidectin susceptibility was only moderately affected. Intestinal Pun-PGP-9 expression elicited a protective effect against ivermectin and moxidectin only in the presence of PP stimulation. Conversely, epidermal Pun-PGP-9 expression protected against moxidectin regardless of PP and against ivermectin only in the absence of active drug ingestion. Our results demonstrate the role of active drug ingestion by nematodes for susceptibility and provide functional evidence for the contribution of P-glycoproteins to ML resistance in a tissue-specific manner. Full article

(This article belongs to the Special Issue Antiparasitics)

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Graphical abstract

Graphical abstract
Full article ">Figure 1
Tissue-specific expression of Pun-PGP-9 in Cel-pgp-9 loss-of-function strain, strain tm830. Verification of transcription and tissue-specific expression. (a) Visible bands for all reverse-transcriptase (RT) PCRs on cDNA templates made from whole-worm total RNA, and no bands for no RT controls and the control strain. (b–e) Confocal microscope images of tissue-specific expression patterns of fixed, freeze cracked, and immunofluorescence-stained transgenic adult Caenorhabditis elegans (blue lookup table (LUT), 405 nM excitation), pharyngeal GFP expression (green LUT, 488 nm excitation) and merged images (DIC +405 nM + 488 nM). Primary antibodies target the FLAG-tag fused to Pun-PGP-9 and the secondary antibody is conjugated with DyLight405. Images were acquired with a confocal Eclipse Ti-U inverted research microscope and processed and merged with ImageJ [<a href="#B40-pharmaceuticals-14-00153" class="html-bibr">40</a>]. All scalebars are 100 µm. (b) Immunostaining in the intestine-Pgp-9 line 1 (white arrows indicate the intestine) and GFP expression at the pharynx. (c) Immunostaining of the epidermis-Pgp-9 strain (white arrows indicate the epidermal syncytium) and pharyngeal GFP expression. (d) Immunostaining in the control strain and pharyngeal GFP expression. (e) Higher magnification of the intestine-Pgp-9 strain (white arrow indicates the apical membrane), Pun-PGP-9: Parascaris univalens P-glycoprotein-9, GFP: green fluorescence protein, DIC: differential interference contrast, int: intestine, vul: vulva, hyp7: hyp7 syncytium, N2∆CelPgp-9 is tm830 (NBRP) [Cel-pgp-9(-)]; Transgenic strains genotypes: Epidermis-Pgp-9 EpiPgp-9Ex1 [Cel-pgp-9(-); Cel-col-19p::Pun-pgp-9::FLAG::Cel-unc-54_3′-UTR; Cel-myo-2p::gfp:: Cel-unc-54_3′UTR]; Intestine-Pgp-9 Line 1 IntPgp-9Ex1 [Cel-pgp-9(-); Cel-ges-1p::Pun-pgp-9::FLAG::Celunc-54_3′-UTR; Cel-myo-2p::gfp::Cel-unc-54_3′UTR]; Control strain CtrlEx1 [Cel-pgp-9(-); Cel-myo-2p::gfp:: Cel-unc-54_3′-UTR]. Full article ">Figure 2
Pharyngeal pumping increases ivermectin and moxidectin susceptibility. Effect of pharyngeal pumping (PP) stimulation by OP50 food bacteria or serotonin in Caenorhabditis elegans on ivermectin and moxidectin susceptibility. (a) Schematic illustration of the experimental setup with active ingestion and intestine drug exposure by PP stimulation. (b) Mean thrashes/minute ± standard error of the mean (SEM) in the negative control (no drug, 1% DMSO) between strains and conditions. Each strain/condition combination was compared to WT OP50− with a Kruskal–Wallis test with Dunn’s post hoc, and p > 0.05 was considered not significant. (c,d) Ivermectin and moxidectin concentration–response curves calculated with GraphPad v8.3.0 based on thrashes/minute in the WT strain with n = 36 per concentration spread equally on three separate days. PP stimulation by OP50 bacteria (OP50+) (black), 4 mM 5-HT (red), or in the absence of a PP stimulus (OP50−) (blue). Significant differences between half-maximal effective concentration (EC50) were calculated using the extra-sum-of-squares-F test and Bonferroni correction. (e) Comparison of the effect of PP stimulation by OP50 Escherichia coli food bacteria (red) or 5-HT (black) in different transgenic and wildtype strains. (c–e) All calculated four parameter non-linear regression models and SEM at each concentration correspond to <a href="#app1-pharmaceuticals-14-00153" class="html-app">Supplementary Tables S1 and S2</a>. Prior to the calculation, the no-drug negative control was set to 0.1 nM and all concentrations were log10 transformed. On the x-axis, the negative control was visualised as “0 M (no drug)” and separated by a break in the axis. P-values < 0.05 were considered significant and are indicated with an asterisk, while corresponding fold-changes are indicated with arrows. 5-HT: serotonin/5-hydroxytryptamine; WT: C. elegans N2 Bristol; Control strain genotype CtrlEx1 [Cel-pgp-9(-); Cel-myo-2p::gfp:: Cel-unc-54_3′-UTR]. Full article ">Figure 3
Modulation of ivermectin and moxidectin susceptibility in C. elegans by transgenic tissue-specific Pun-PGP-9 expression. (a–d) Concentration–response curves corresponding to <a href="#app1-pharmaceuticals-14-00153" class="html-app">Tables S1 and S2</a> for ivermectin (a/c) and moxidectin (b/d) in the absence of a PP stimulus (OP50−) or PP stimulation by serotonin (5-HT+/OP50-) in liquid S-medium. (a–d) show the control strain’s response for 5-HT+ (black dashed line with triangles) or OP50− (grey dashed line with circles), (a,b) show the epidermis-Pgp-9 strain for 5-HT+ (dark blue with open squares) or OP50− (orange with rectangles) and (c,d) show the intestine-Pgp-9 line 1 for 5-HT+ (light blue with open squares) or OP50− (red with rectangles). All concentration–response curves are based on four-parameter non-linear regression models calculated from the motility response (thrashes/minute) of 36 synchronised 1-day adults per concentration, and error bars represent standard error of the mean. Prior to calculation, concentrations were log10 transformed, and the no-drug negative control was set to 0.1 nM. On the x-axis, the negative control was visualised as “0 M (no drug)” and separated by a break in the axis. Significant differences in EC50 values were compared using the extra-sum-of-squares-F test and Bonferroni correction; p-values < 0.05 were considered significant and indicated with an asterisk (*), while corresponding fold-changes are indicated with arrows. Pun-PGP-9: Parascaris univalens P-glycoprotein-9; N2∆CelPgp-9 is tm830 (NBRP) [Cel-pgp-9(-)]; Transgenic strains genotypes: Epidermis-Pgp-9 EpiPgp-9Ex1 [Cel-pgp-9(-); Cel-col-19p::Pun-pgp-9::FLAG::Cel-unc-54_3′-UTR; Cel-myo-2p::gfp:: Cel-unc-54_3′UTR]; Intestine-Pgp-9 Line 1 IntPgp-9Ex1 [Cel-pgp-9(-); Cel-ges-1p::Pun-pgp-9::FLAG::Celunc-54_3′-UTR; Cel-myo-2p::gfp::Cel-unc-54_3′UTR]; Control strain CtrlEx1 [Cel-pgp-9(-); Cel-myo-2p::gfp:: Cel-unc-54_3′-UTR]. Full article ">Figure 4
Comparison of moxidectin and ivermectin in wildtype and transgenic Caenorhabditis elegans strains. Forrest plot visualising the EC50 and corresponding 95% confidence intervals for ivermectin (red) and moxidectin (turquoise) in transgenic and wildtype Caenorhabditis elegans strains in the presence of pharyngeal pumping (PP) stimulation by OP50 food bacteria (OP50+) or 5-hydroxytryptamine (5-HT+) or no PP stimulation (OP50−) were visualised using ggplot2 [<a href="#B42-pharmaceuticals-14-00153" class="html-bibr">42</a>] in R v4.0.3 [<a href="#B43-pharmaceuticals-14-00153" class="html-bibr">43</a>]. EC50 values were inferred from four-parameter linear regression models calculated from 36 synchronised 1-day adult worms per concentration, strain, and condition using GraphPad v8.3.0. Worms were incubated for 24 h in S-medium containing a concentration series of ivermectin or moxidectin in a final DMSO concentration of 1%. Epidermis-Pgp-9 genotype is EpiPgp-9Ex1 [Cel-pgp-9(-); Cel-col-19p::Pun-pgp-9::FLAG::Cel-unc-54_3′-UTR; Cel-myo-2p::gfp:: Cel-unc-54_3′UTR];. Intestine Pgp-9 Line 1 (L1) and Line 2 (L2) genotype is IntPgp-9Ex1 or 2 [Cel-pgp-9(-); Cel-ges-1p::Pun-pgp-9::FLAG::Celunc-54_3′-UTR; Cel-myo-2p::gfp::Cel-unc-54_3′UTR];. Control strain genotype is CtrlEx1 [Cel-pgp-9(-); Cel-myo-2p::gfp::Cel-unc-54_3′-UTR]; WT is N2 Bristol. Pgp: P-glycoprotein; Pun-PGP-9: Parascaris univalens P-glycoprotein-9; IVM: ivermectin; MOX: moxidectin; PP: pharyngeal pumping; WT: wildtype. Full article ">Figure 5
Schematic illustration of P-glycoprotein-mediated barrier function. Hypothetical schematic illustration of Pgp-mediated barrier function in a Caenorhabditis elegans adult. Expression of P-glycoproteins in specific barrier tissues, i.e., the epidermis and the intestine prohibit MLs from reaching target tissues, thereby preventing an ML-induced hyperpolarisation of the neurons and muscle paralysis. Full article ">

14 pages, 824 KiB

Open AccessReview

Therapeutic Applications of Type 2 Diabetes Mellitus Drug Metformin in Patients with Osteoarthritis

by Parkyong Song, Ji Sun Hwang, Hyean Cheal Park, Keun Ki Kim, Hong-Joo Son, Yu-Jin Kim and Kwang Min Lee

Pharmaceuticals 2021, 14(2), 152; https://doi.org/10.3390/ph14020152 - 13 Feb 2021

Cited by 13 | Viewed by 3657

Abstract

Type 2 diabetes mellitus (T2DM) and osteoarthritis (OA) are common chronic diseases that frequently co-exist. The link between OA and T2DM is attributed to common risk factors, including age and obesity. Several reports suggest that hyperglycemia and accumulated advanced glycosylation end-products might regulate [...] Read more.

Type 2 diabetes mellitus (T2DM) and osteoarthritis (OA) are common chronic diseases that frequently co-exist. The link between OA and T2DM is attributed to common risk factors, including age and obesity. Several reports suggest that hyperglycemia and accumulated advanced glycosylation end-products might regulate cartilage homeostasis and contribute to the development and progression of OA. Metformin is used widely as the first-line treatment for T2DM. The drug acts by regulating glucose levels and improving insulin sensitivity. The anti-diabetic effects of metformin are mediated mainly via activation of adenosine monophosphate (AMP)-activated protein kinase (AMPK), which is an energy sensing enzyme activated directly by an increase in the AMP/ATP ratio under conditions of metabolic stress. Dysregulation of AMPK is strongly associated with development of T2DM and metabolic syndrome. In this review, we discuss common risk factors, the association between OA and T2DM, and the role of AMPK. We also address the adaptive use of metformin, a known AMPK activator, as a new drug for treatment of patients with OA and T2DM. Full article

(This article belongs to the Special Issue Drug and Therapy for Osteoarthritis (OA))

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18 pages, 982 KiB

Open AccessReview

Immune Checkpoint Inhibition in Oesophago-Gastric Carcinoma

by Anica Högner and Peter Thuss-Patience

Pharmaceuticals 2021, 14(2), 151; https://doi.org/10.3390/ph14020151 - 12 Feb 2021

Cited by 18 | Viewed by 5126

Abstract

Immune checkpoint inhibitors enrich the therapeutic landscape in oesophago-gastric carcinoma. With regard to oesophageal squamous cell carcinoma (ESCC), the selective PD-1 (programmed cell death receptor 1)-inhibitor nivolumab improves disease-free survival in the adjuvant therapy setting (CHECKMATE-577). In first-line treatment, ESCC patients (pts) benefit [...] Read more.

Immune checkpoint inhibitors enrich the therapeutic landscape in oesophago-gastric carcinoma. With regard to oesophageal squamous cell carcinoma (ESCC), the selective PD-1 (programmed cell death receptor 1)-inhibitor nivolumab improves disease-free survival in the adjuvant therapy setting (CHECKMATE-577). In first-line treatment, ESCC patients (pts) benefit in overall survival (OS) from the PD-1-inhibitor pembrolizumab in combination with chemotherapy (KEYNOTE-590). In the second-line setting, nivolumab (ATTRACTION-03) and pembrolizumab (KEYNOTE-181) demonstrate a benefit in OS compared with chemotherapy. These data resulted in the approval of nivolumab for the second-line treatment of advanced ESCC pts regardless of PD-L1 (programmed cell death ligand 1) status in Europe, Asia, and the USA, and pembrolizumab for pts with PD-L1 CPS (combined positivity score) ≥ 10 in Asia and the USA. Further approvals can be expected. In gastro-oesophageal junction and gastric cancer, the addition of nivolumab to chemotherapy in first-line treatment improves OS in pts with advanced disease with PD-L1 CPS ≥ 5 (CHECKMATE-649). Additionally, pembrolizumab was non-inferior to chemotherapy for OS in PD-L1 CPS ≥ 1 pts (KEYNOTE-062). In third-line treatment, nivolumab shows benefits in OS regardless of PD-L1 expression (ATTRACTION-02) with approval in Asia, and pembrolizumab prolonged the duration of response in PD-L1 positive pts (KEYNOTE-059) with approval in the USA. We discuss the recent results of the completed phase II and III clinical trials. Full article

(This article belongs to the Special Issue Immune Checkpoint Inhibitor Therapy)

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13 pages, 379 KiB

Open AccessArticle

Assessment of Novel Inhaler Technique Reminder Labels in Image Format on the Correct Demonstration of Inhaler Technique Skills in Asthma: A Single-Blinded Randomized Controlled Trial

by Iman Basheti, Bassam Mahboub, Laila Salameh, Mena Al-Ani, Ammar Abdulrahman Jairoun, Basema Saddik and Eman Abu-Gharbieh

Pharmaceuticals 2021, 14(2), 150; https://doi.org/10.3390/ph14020150 - 12 Feb 2021

Cited by 4 | Viewed by 2268

Abstract

Background: Prevalence of asthma in the United Arab Emirates (UAE) is high, and training patients on correct inhaler technique is vital. Objectives: To assess the effectiveness of inhaler technique labels incorporating the individual technique steps in image format on the retention of correct [...] Read more.

Background: Prevalence of asthma in the United Arab Emirates (UAE) is high, and training patients on correct inhaler technique is vital. Objectives: To assess the effectiveness of inhaler technique labels incorporating the individual technique steps in image format on the retention of correct inhaler technique for patients with asthma living in the UAE and following inhaler training; secondly to investigate the effect of inhaler technique education using self-check pictorial labels on patients’ overall asthma control. Methods: This single-blinded randomized controlled study was conducted in 2019 and followed consecutive recruitment of asthma patients visiting respiratory clinics at Rashid Hospital in Dubai. Patients were using a controller inhaler (Turbuhaler (TH), Accuhaler (ACC), or pressurized metered-dose inhaler (pMDI)). Following recruitment, patients were randomized into active group receiving educational intervention plus the inhaler label, and control group receiving educational intervention without the label. Patients were assessed at baseline and at one-month on their inhaler technique and asthma control. Results: Participants (n = 245; 93 = TH, 70 = ACC, 82 = pMDI) showed a significant difference between the groups at one-month for inhaler technique scores for TH (active 5.29 ± 1.86 vs. control = 24.4 ± 21.28), ACC (active = 3.99 ± 1.43 vs. control = 25.45 ± 22.57), and pMDI (active = 4.59 ± 0.10 vs. control = 120.55 ± 17.2), p < 0.001 for all. Asthma control for active group indicated significant improvements compared to control for TH and pMDI (p < 0.001 for both), but not ACC group (p = 0.087). Conclusions: Retention of correct inhaler technique and improved asthma control can be enhanced by using a specialized inhaler technique label in image format. Full article

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23 pages, 3860 KiB

Open AccessReview

The Role of miRNA-7 in the Biology of Cancer and Modulation of Drug Resistance

by Ewa Gajda, Małgorzata Grzanka, Marlena Godlewska and Damian Gawel

Pharmaceuticals 2021, 14(2), 149; https://doi.org/10.3390/ph14020149 - 12 Feb 2021

Cited by 16 | Viewed by 3409

Abstract

MicroRNAs (miRNAs, miRs) are small non-coding RNA (ncRNA) molecules capable of regulating post-transcriptional gene expression. Imbalances in the miRNA network have been associated with the development of many pathological conditions and diseases, including cancer. Recently, miRNAs have also been linked to the phenomenon [...] Read more.

MicroRNAs (miRNAs, miRs) are small non-coding RNA (ncRNA) molecules capable of regulating post-transcriptional gene expression. Imbalances in the miRNA network have been associated with the development of many pathological conditions and diseases, including cancer. Recently, miRNAs have also been linked to the phenomenon of multidrug resistance (MDR). MiR-7 is one of the extensively studied miRNAs and its role in cancer progression and MDR modulation has been highlighted. MiR-7 is engaged in multiple cellular pathways and acts as a tumor suppressor in the majority of human neoplasia. Its depletion limits the effectiveness of anti-cancer therapies, while its restoration sensitizes cells to the administered drugs. Therefore, miR-7 might be considered as a potential adjuvant agent, which can increase the efficiency of standard chemotherapeutics. Full article

(This article belongs to the Special Issue MiRNA-Based Therapeutics in Cancer)

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46 pages, 6710 KiB

Open AccessReview

Modulation of the Serotonergic Receptosome in the Treatment of Anxiety and Depression: A Narrative Review of the Experimental Evidence

by Gustavo R. Villas-Boas, Stefânia N. Lavorato, Marina M. Paes, Pablinny M. G. de Carvalho, Vanessa C. Rescia, Mila S. Cunha, Manoel F. de Magalhães-Filho, Luis F. Ponsoni, Adryano Augustto Valladao de Carvalho, Roseli B. de Lacerda, Lais da S. Leite, Matheus da S. Tavares-Henriques, Luiz A. F. Lopes, Luiz G. R. Oliveira, Saulo E. Silva-Filho, Ana P. S. da Silveira, Roberto K. N. Cuman, Francielli M. de S. Silva-Comar, Jurandir F. Comar, Luana do A. Brasileiro, Jussileide N. dos Santos, William R. de Freitas, Katyuscya V. Leão, Jonatas G. da Silva, Raphael C. Klein, Mary H. F. Klein, Bruno H. da S. Ramos, Cristiane K. C. Fernandes, Dayane G. de L. Ribas and Silvia A. Oesterreich Show full author list Hide full author list

Pharmaceuticals 2021, 14(2), 148; https://doi.org/10.3390/ph14020148 - 12 Feb 2021

Cited by 20 | Viewed by 6592

Abstract

Serotonin (5-HT) receptors are found throughout central and peripheral nervous systems, mainly in brain regions involved in the neurobiology of anxiety and depression. 5-HT receptors are currently promising targets for discovering new drugs for treating disorders ranging from migraine to neuropsychiatric upsets, such [...] Read more.

Serotonin (5-HT) receptors are found throughout central and peripheral nervous systems, mainly in brain regions involved in the neurobiology of anxiety and depression. 5-HT receptors are currently promising targets for discovering new drugs for treating disorders ranging from migraine to neuropsychiatric upsets, such as anxiety and depression. It is well described in the current literature that the brain expresses seven types of 5-HT receptors comprising eighteen distinct subtypes. In this article, we comprehensively reviewed 5-HT1-7 receptors. Of the eighteen 5-HT receptors known today, thirteen are G protein-coupled receptors (GPCRs) and represent targets for approximately 40% of drugs used in humans. Signaling pathways related to these receptors play a crucial role in neurodevelopment and can be modulated to develop effective therapies to treat anxiety and depression. This review presents the experimental evidence of the modulation of the “serotonergic receptosome” in the treatment of anxiety and depression, as well as demonstrating state-of-the-art research related to phytochemicals and these disorders. In addition, detailed aspects of the pharmacological mechanism of action of all currently known 5-HT receptor families were reviewed. From this review, it will be possible to direct the rational design of drugs towards new therapies that involve signaling via 5-HT receptors. Full article

(This article belongs to the Special Issue Molecular Pharmacology of 5-HT Receptors)

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17 pages, 1448 KiB

Open AccessArticle

Application of RP-18 TLC Retention Data to the Prediction of the Transdermal Absorption of Drugs

by Anna W. Sobańska, Jeremy Robertson and Elżbieta Brzezińska

Pharmaceuticals 2021, 14(2), 147; https://doi.org/10.3390/ph14020147 - 12 Feb 2021

Cited by 13 | Viewed by 2596

Abstract

Several chromatographic parameters (R_M⁰ and S obtained from RP-18 TLC with methanol—pH 7.4 phosphate buffer mobile phases by extrapolation to zero concentration of methanol; R_f and R_M obtained from RP-18 TLC with acetonitrile—pH 7.4 phosphate buffer 70:30 v [...] Read more.

Several chromatographic parameters (R_M⁰ and S obtained from RP-18 TLC with methanol—pH 7.4 phosphate buffer mobile phases by extrapolation to zero concentration of methanol; R_f and R_M obtained from RP-18 TLC with acetonitrile—pH 7.4 phosphate buffer 70:30 v/v as a mobile phase) and calculated molecular descriptors (molecular weight—M_W; molar volume—V_M; polar surface area—PSA; total count of nitrogen and oxygen atoms—(N+O); H-bond donor count—HD; H-bond acceptor count—HA; distribution coefficient—log D; total energy—E_T; binding energy—E_b; hydration energy—E_h; energy of the highest occupied molecular orbital—E_HOMO; energy of the lowest unoccupied orbital—E_LUMO; electronic energy—E_e; surface area—S_a; octanol-water partition coefficient—log P; dipole moment—DM; refractivity—R, polarizability—α) and their combinations (R_f/PSA, R_M/M_W, R_M/V_M) were tested in order to generate useful models of solutes’ skin permeability coefficient log K_p. It was established that neither R_M⁰ nor S obtained in the conditions used in this study is a good predictor of the skin permeability coefficient. The chromatographic parameters R_f and R_f/PSA were also unsuitable for this purpose. A simple and potentially useful, purely computational model based on (N+O), log D and HD as independent variables and accounting for ca. 83% of total variability was obtained. The evaluation of parameters derived from R_M (R_M, R_M/M_W, R_M/V_M) as independent variables in log K_p models proved that R_M/V_M is the most suitable descriptor belonging to this group. In a search for a reliable log K_p model based on this descriptor two possibilities were considered: a relatively simple model based on 5 independent variables: (N+O), log D, R_M/V_M, E_T and E_h and a more complex one, involving also E_b, M_W and PSA. Full article

(This article belongs to the Special Issue Selected Papers from the 6th International Electronic Conference on Medicinal Chemistry (ECMC2020))

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26 pages, 7174 KiB

Open AccessArticle

Zingerone Targets Status Epilepticus by Blocking Hippocampal Neurodegeneration via Regulation of Redox Imbalance, Inflammation and Apoptosis

by Summya Rashid, Adil Farooq Wali, Shahzada Mudasir Rashid, Rana M. Alsaffar, Ajaz Ahmad, Basit L. Jan, Bilal Ahmad Paray, Saeed M. A. Alqahtani, Azher Arafah and Muneeb U. Rehman

Pharmaceuticals 2021, 14(2), 146; https://doi.org/10.3390/ph14020146 - 11 Feb 2021

Cited by 23 | Viewed by 3366

Abstract

Epilepsy is an intricate neurological disease where the neurons are severely affected, leading to the mortality of millions worldwide. Status epilepticus (SE), induced by lithium chloride (LiCl) and pilocarpine, is the most accepted model for epilepsy. The current work aims to unravel the [...] Read more.

Epilepsy is an intricate neurological disease where the neurons are severely affected, leading to the mortality of millions worldwide. Status epilepticus (SE), induced by lithium chloride (LiCl) and pilocarpine, is the most accepted model for epilepsy. The current work aims to unravel the mechanisms underlying the anti-epileptic efficacy of zingerone (an active ingredient of ginger), which has beneficial pharmacological activities on seizure-induced behavioral, histological, neurochemical, and molecular patterns in mice. Zingerone restored cognitive function by diminishing seizure activity, escape latency, and subsequent hippocampal damage manifested in histology. Seizures are associated with local inflammation, redox imbalance, and neural loss, confirmed by the present study of SE, and was attenuated by zingerone treatment. Nuclear factor-kappa B and its downstream signaling molecules (TNF-α, IL-1β, IL-6, NO, MPO) were activated in the LiCl-and-pilocarpine-induced group leading to inflammatory signaling, which was substantially ameliorated by zingerone treatment. The intrinsic apoptotic process was triggered subsequent to SE, as demonstrated by augmentation of cleaved caspase-3, downregulation of Bcl-2. However, zingerone treatment downregulated caspase-3 and upregulated Bcl-2, increasing cell survival and decreasing hippocampal neural death, deciphering involvement of apoptosis in SE. Therefore, zingerone plays an essential role in neuroprotection, probably by precluding oxidative stress, inflammation, and obstructing the mitochondrial pathway of apoptosis. Full article

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

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14 pages, 1726 KiB

Open AccessFeature PaperEditor’s ChoiceReview

2020 FDA TIDES (Peptides and Oligonucleotides) Harvest

by Othman Al Musaimi, Danah Al Shaer, Fernando Albericio and Beatriz G. de la Torre

Pharmaceuticals 2021, 14(2), 145; https://doi.org/10.3390/ph14020145 - 11 Feb 2021

Cited by 56 | Viewed by 5912

Abstract

2020 has been an extremely difficult and challenging year as a result of the coronavirus disease 2019 (COVID-19) pandemic and one in which most efforts have been channeled into tackling the global health crisis. The US Food and Drug Administration (FDA) has approved [...] Read more.

2020 has been an extremely difficult and challenging year as a result of the coronavirus disease 2019 (COVID-19) pandemic and one in which most efforts have been channeled into tackling the global health crisis. The US Food and Drug Administration (FDA) has approved 53 new drug entities, six of which fall in the peptides and oligonucleotides (TIDES) category. The number of authorizations for these kinds of drugs has been similar to that of previous years, thereby reflecting the consolidation of the TIDES market. Here, the TIDES approved in 2020 are analyzed in terms of chemical structure, medical target, mode of action, and adverse effects. Full article

(This article belongs to the Special Issue The Story of Successful Drugs and Recent FDA-Approved Molecules)

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A total of 228 new drugs were approved by the Food and Drug Administration (FDA) from 2016 to 2020 [<a href="#B2-pharmaceuticals-14-00145" class="html-bibr">2</a>]. mAbs, monoclonal antibodies; ADCs, antibody drug conjugates; Oligos, oligonucleotides. Full article ">Figure 2
Structure of viltolarsen (ViltepsoTM). Full article ">Figure 3
Chemical structure of lumasiran (OxlumoTM). Full article ">Figure 4
Chemical structure of setmelanotide (ImcivreeTM). Full article ">Figure 5
Chemical structure of 64Cu -DOTATATE (DetectnetTM). Full article ">Figure 6
Chemical structure of 68Ga-PMSA-11. Full article ">Figure 7
Chemical structure of belantamab mafodotin-blmf (BlenrepTM). Full article ">Figure 8
Chemical structure of: (A) natural dolastatin 10; (B) monomethyl auristatin F (MMAF); and (C) monomethyl auristatin E (MMAE). Differences from the natural molecule are shown in red [<a href="#B44-pharmaceuticals-14-00145" class="html-bibr">44</a>]. Full article ">

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Pharmaceuticals, Volume 14, Issue 2 (February 2021) – 100 articles

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