Next-generation sequencing (NGS) has transitioned from research to clinical routine, yet the comparability of different technologies for mutation profiling remains an open question. We performed a European multicenter (n=6) evaluation of three amplicon-based NGS assays targeting 11 genes recurrently mutated in chronic lymphocytic leukemia. Each assay was assessed by two centers using 48 pre-characterized chronic lymphocytic leukemia samples; libraries were sequenced on the Illumina MiSeq instrument and bioinformatics analyses were centralized. Across all centers the median percentage of target reads ≥100x ranged from 94.2- 99.8%. In order to rule out assay-specific technical variability, we first assessed variant calling at the individual assay level i.e., pairwise analysis of variants detected amongst partner centers. After filtering for variants present in the paired normal sample and removal of PCR/sequencing artefacts, the panels achieved 96.2% (Multiplicom), 97.7% (TruSeq) and 90% (HaloPlex) concordance at a variant allele frequency (VAF) >0.5%. Reproducibility was assessed by looking at the inter-laboratory variation in detecting mutations and 107 of 115 (93% concordance) mutations were detected by all six centers, while the remaining eight variants (7%) were undetected by a single center. Notably, 6 of 8 of these variants concerned minor subclonal mutations (VAF <5%). We sought to investigate low-frequency mutations further by using a high-sensitivity assay containing unique molecular identifiers, which confirmed the presence of several minor subclonal mutations. Thus, while amplicon-based approaches can be adopted for somatic mutation detection with VAF >5%, after rigorous validation, the use of unique molecular identifiers may be necessary to reach a higher sensitivity and ensure consistent and accurate detection of low-frequency variants.
The therapeutic potential of a novel, targeted-release formulation of oral budesonide (Nefecon) for the treatment of IgA nephropathy (IgAN) was first demonstrated by the phase 2b NEFIGAN trial. To verify these findings, the phase 3 NefigArd trial tested the efficacy and safety of nine months of treatment with Nefecon (16 mg/d) versus placebo in adult patients with primary IgAN at risk of progressing to kidney failure (ClinicalTrials.gov: NCT03643965). NefIgArd was a multicenter, randomized, double-blind, placebo-controlled two-part trial. In Part A, 199 patients with IgAN were treated with Nefecon or placebo for nine months and observed for an additional three months. The primary endpoint for Part A was 24-hour urine protein-to-creatinine ratio (UPCR) after nine months. Secondary efficacy outcomes evaluated included estimated glomerular filtration rate (eGFR) at nine and 12 months and the UPCR at 12 months. At nine months, UPCR was 27% lower in the Nefecon group compared with placebo, along with a benefit in eGFR preservation corresponding to a 3.87 ml/min/1.73 m2 difference versus placebo (both significant). Nefecon was well-tolerated, and treatment-emergent adverse events were mostly mild to moderate in severity and reversible. Part B is ongoing and will be reported on later. Thus, NefIgArd is the first phase 3 IgA nephropathy trial to show clinically important improvements in UPCR and eGFR and confirms the findings from the phase 2b NEFIGAN study.
- Keywords
- IgA nephropathy, glomerular disease, glucocorticoids, gut-associated lymphoid tissue,
- MeSH
- Budesonide * administration & dosage MeSH
- Adult MeSH
- Double-Blind Method MeSH
- Glomerular Filtration Rate MeSH
- Glomerulonephritis, IGA * drug therapy MeSH
- Humans MeSH
- Kidney Function Tests MeSH
- Treatment Outcome MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Publication type
- Journal Article MeSH
- Multicenter Study MeSH
- Randomized Controlled Trial MeSH
- Names of Substances
- Budesonide * MeSH
Targeted therapies and the consequent adoption of "personalized" oncology have achieved notable successes in some cancers; however, significant problems remain with this approach. Many targeted therapies are highly toxic, costs are extremely high, and most patients experience relapse after a few disease-free months. Relapses arise from genetic heterogeneity in tumors, which harbor therapy-resistant immortalized cells that have adopted alternate and compensatory pathways (i.e., pathways that are not reliant upon the same mechanisms as those which have been targeted). To address these limitations, an international task force of 180 scientists was assembled to explore the concept of a low-toxicity "broad-spectrum" therapeutic approach that could simultaneously target many key pathways and mechanisms. Using cancer hallmark phenotypes and the tumor microenvironment to account for the various aspects of relevant cancer biology, interdisciplinary teams reviewed each hallmark area and nominated a wide range of high-priority targets (74 in total) that could be modified to improve patient outcomes. For these targets, corresponding low-toxicity therapeutic approaches were then suggested, many of which were phytochemicals. Proposed actions on each target and all of the approaches were further reviewed for known effects on other hallmark areas and the tumor microenvironment. Potential contrary or procarcinogenic effects were found for 3.9% of the relationships between targets and hallmarks, and mixed evidence of complementary and contrary relationships was found for 7.1%. Approximately 67% of the relationships revealed potentially complementary effects, and the remainder had no known relationship. Among the approaches, 1.1% had contrary, 2.8% had mixed and 62.1% had complementary relationships. These results suggest that a broad-spectrum approach should be feasible from a safety standpoint. This novel approach has potential to be relatively inexpensive, it should help us address stages and types of cancer that lack conventional treatment, and it may reduce relapse risks. A proposed agenda for future research is offered.
- Keywords
- Cancer hallmarks, Integrative medicine, Multi-targeted, Phytochemicals, Targeted therapy,
- MeSH
- Drug Resistance, Neoplasm genetics MeSH
- Molecular Targeted Therapy * MeSH
- Antineoplastic Agents, Phytogenic therapeutic use MeSH
- Genetic Heterogeneity * MeSH
- Precision Medicine * MeSH
- Humans MeSH
- Tumor Microenvironment genetics MeSH
- Neoplasms genetics pathology prevention & control therapy MeSH
- Signal Transduction MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Review MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, N.I.H., Intramural MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Names of Substances
- Antineoplastic Agents, Phytogenic MeSH
Resveratrol and other natural stilbenoids, including piceatannol, pterostilbene, and gnetol, are well-known anti-inflammatory compounds with indisputable activity in vitro as well as in vivo. Their molecular targets include inducible nitric oxide synthase, cyclooxygenases, leukotrienes, nuclear factor kappa B, tumor necrosis factor α, interleukins and many more. This anti-inflammatory activity together with their antioxidant activity is believed to stand behind their other positive health effects against cancer, cardiovascular and neurodegenerative diseases or diabetes. Thus, they are nowadays commercially marketed as nutraceuticals. Naturally, they are present in wine, grapes or berries. However, there is a rigorous debate about the real effect of these compounds on human health. It is argued that the concentration of stilbenoids in food and beverages is too low to have any therapeutic potential and this concentration is further reduced by their low bioavailability and extensive metabolism. Therefore, this review focuses on in vitro, in vivo, preclinical as well as clinical data available for various natural stilbenoids and summarizes the anti-inflammatory targets on molecular level, compares the relevance of the experimental studies, discusses the metabolism of stilbenoids and the potential activity of their metabolites and relates this knowledge to human health. Moreover, the ways to augment stilbenoidś efficacy are suggested with special focus on multitargeted therapy and nanocarriers.
- Keywords
- Bioavailability, Encapsulation, Inflammation, Metabolites, Multi-targeted therapy, Natural stilbenes,
- MeSH
- Anti-Inflammatory Agents * pharmacokinetics pharmacology therapeutic use MeSH
- Humans MeSH
- Stilbenes * pharmacokinetics pharmacology therapeutic use MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Review MeSH
- Names of Substances
- Anti-Inflammatory Agents * MeSH
- Stilbenes * MeSH
Alzheimer's disease (AD) has a complex pathophysiology that includes aggregation of pathological proteins, impaired neurotransmission, increased oxidative stress, or microglia-mediated neuroinflammation. Therapeutics targeting only one of these AD-related subpathologies have not yet been successful in the search for a disease-modifying treatment. Therefore, multi-target drugs (MTDs) aiming simultaneously at several subpathologies are expected to be a better approach. However, the concept of MTD is inherently connected with several limitations, which are often ignored during MTD design and development. Here, we provide an overview of the MTD approach and discuss its potential pitfalls in the context of AD treatment. We also put forward ideas to be used in the rational design of MTDs to obtain drugs that are effective against AD.
- Keywords
- Alzheimer’s disease, designed multiple ligands, drug design, multi-target drugs, polypharmacology,
- MeSH
- Alzheimer Disease * drug therapy MeSH
- Pharmaceutical Preparations * MeSH
- Drug Delivery Systems MeSH
- Humans MeSH
- Synaptic Transmission MeSH
- Oxidative Stress MeSH
- Inflammation MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Review MeSH
- Names of Substances
- Pharmaceutical Preparations * MeSH
Thanks to the widespread use and safety profile of donepezil (1) in the treatment of Alzheimer's disease (AD), one of the most widely adopted multi-target-directed ligand (MTDL) design strategies is to modify its molecular structure by linking a second fragment carrying an additional AD-relevant biological property. Herein, supported by a proposed combination therapy of 1 and the quinone drug idebenone, we rationally designed novel 1-based MTDLs targeting Aβ and oxidative pathways. By exploiting a bioisosteric replacement of the indanone core of 1 with a 1,4-naphthoquinone, we ended up with a series of highly merged derivatives, in principle devoid of the "physicochemical challenge" typical of large hybrid-based MTDLs. A preliminary investigation of their multi-target profile identified 9, which showed a potent and selective butyrylcholinesterase inhibitory activity, together with antioxidant and antiaggregating properties. In addition, it displayed a promising drug-like profile.
- Keywords
- Alzheimer's disease, drug design, medicinal chemistry, multi-target drug discovery, polypharmacology,
- MeSH
- Acetylcholinesterase chemistry metabolism MeSH
- Alzheimer Disease drug therapy MeSH
- Amyloid beta-Peptides antagonists & inhibitors metabolism MeSH
- Antioxidants chemistry metabolism pharmacology MeSH
- Cholinesterase Inhibitors chemistry metabolism pharmacology therapeutic use MeSH
- Donepezil chemistry metabolism pharmacology therapeutic use MeSH
- Blood-Brain Barrier diagnostic imaging metabolism MeSH
- Indans chemistry MeSH
- Humans MeSH
- Ligands * MeSH
- Cell Line, Tumor MeSH
- Neuroprotective Agents chemistry metabolism pharmacology therapeutic use MeSH
- Oxidative Stress drug effects MeSH
- Protein Aggregates drug effects MeSH
- Drug Design MeSH
- Cell Survival drug effects MeSH
- Structure-Activity Relationship MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Acetylcholinesterase MeSH
- Amyloid beta-Peptides MeSH
- Antioxidants MeSH
- Cholinesterase Inhibitors MeSH
- Donepezil MeSH
- indacrinone MeSH Browser
- Indans MeSH
- Ligands * MeSH
- Neuroprotective Agents MeSH
- Protein Aggregates MeSH
Multidrug resistance (MDR) contributes to failure of chemotherapy. We here show that biodegradable polymeric nanogels are able to overcome MDR via folic acid targeting. The nanogels are based on hydroxyethyl methacrylamide-oligoglycolates-derivatized poly(hydroxyethyl methacrylamide-co-N-(2-azidoethyl)methacrylamide) (p(HEMAm-co-AzEMAm)-Gly-HEMAm), covalently loaded with the chemotherapeutic drug doxorubicin (DOX) and subsequently decorated with a folic acid-PEG conjugate via copper-free click chemistry. pH-Responsive drug release is achieved via the acid-labile hydrazone bond between DOX and the methacrylamide polymeric network. Cellular uptake and cytotoxicity analyses in folate receptor-positive B16F10 melanoma versus folate receptor-negative A549 lung carcinoma cells confirmed specific uptake of the targeted nanogels. Confocal microscopy demonstrated efficient internalization, lysosomal trafficking, drug release and nuclear localization of DOX. We also show that DOX resistance in 4T1 breast cancer cells results in upregulation of the folate receptor, and that folic acid targeted nanogels can be employed to bypass drug efflux pumps, resulting in highly efficient killing of resistant cancer cells. In conclusion, folic acid functionalized nanogels with pH-controlled drug release seem to hold significant potential for treating multidrug resistant malignancies.
- MeSH
- A549 Cells MeSH
- Drug Resistance, Neoplasm * MeSH
- Molecular Targeted Therapy MeSH
- Doxorubicin administration & dosage MeSH
- Folate Receptors, GPI-Anchored metabolism MeSH
- Hydrogen-Ion Concentration MeSH
- Drug Delivery Systems * MeSH
- Humans MeSH
- Melanoma, Experimental MeSH
- Drug Resistance, Multiple * MeSH
- Cell Line, Tumor MeSH
- Nanoparticles * MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Doxorubicin MeSH
- Folate Receptors, GPI-Anchored MeSH
The multifactorial nature of Alzheimer's disease (AD) is a reason for the lack of effective drugs as well as a basis for the development of "multi-target-directed ligands" (MTDLs). As cases increase in developing countries, there is a need of new drugs that are not only effective but also accessible. With this motivation, we report the first sustainable MTDLs, derived from cashew nutshell liquid (CNSL), an inexpensive food waste with anti-inflammatory properties. We applied a framework combination of functionalized CNSL components and well-established acetylcholinesterase (AChE)/butyrylcholinesterase (BChE) tacrine templates. MTDLs were selected based on hepatic, neuronal, and microglial cell toxicity. Enzymatic studies disclosed potent and selective AChE/BChE inhibitors (5, 6, and 12), with subnanomolar activities. The X-ray crystal structure of 5 complexed with BChE allowed rationalizing the observed activity (0.0352 nM). Investigation in BV-2 microglial cells revealed antineuroinflammatory and neuroprotective activities for 5 and 6 (already at 0.01 μM), confirming the design rationale.
- MeSH
- Acetylcholinesterase chemistry metabolism MeSH
- Alzheimer Disease drug therapy pathology MeSH
- Anacardium chemistry metabolism MeSH
- Cell Line MeSH
- Butyrylcholinesterase chemistry metabolism MeSH
- Cytokines metabolism MeSH
- Catalytic Domain MeSH
- Humans MeSH
- Ligands * MeSH
- Lipopolysaccharides pharmacology MeSH
- Microglia cytology drug effects metabolism MeSH
- Neuroprotective Agents chemistry metabolism pharmacology therapeutic use MeSH
- Nuts chemistry metabolism MeSH
- Drug Design MeSH
- Plant Extracts chemistry MeSH
- Molecular Dynamics Simulation MeSH
- Tacrine chemistry metabolism MeSH
- Binding Sites MeSH
- Cell Survival drug effects MeSH
- Structure-Activity Relationship MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Acetylcholinesterase MeSH
- Butyrylcholinesterase MeSH
- Cytokines MeSH
- Ligands * MeSH
- Lipopolysaccharides MeSH
- Neuroprotective Agents MeSH
- Plant Extracts MeSH
- Tacrine MeSH
Multi-target drug discovery is one of the most followed approaches in the active central nervous system (CNS) therapeutic area, especially in the search for new drugs against Alzheimer's disease (AD). This is because innovative multi-target-directed ligands (MTDLs) could more adequately address the complexity of this pathological condition. In a continuation of our efforts aimed at a new series of anti-AD MTDLs, we combined the structural features of the cholinesterase inhibitor drug tacrine with that of resveratrol, which is known for its purported antioxidant and anti-neuroinflammatory activities. The most interesting hybrid compounds (5, 8, 9 and 12) inhibited human acetylcholinesterase at micromolar concentrations and effectively modulated Aβ self-aggregation in vitro. In addition, 12 showed intriguing anti-inflammatory and immuno-modulatory properties in neuronal and glial AD cell models. Importantly, the MTDL profile is accompanied by high-predicted blood-brain barrier permeability, and low cytotoxicity on primary neurons.
- Keywords
- Acetylcholinesterase, Alzheimer's disease, Amyloid, Multitarget compounds, Neuroinflammation,
- MeSH
- Acetylcholinesterase metabolism MeSH
- Alzheimer Disease drug therapy metabolism MeSH
- Amyloid beta-Peptides chemistry MeSH
- Antioxidants chemistry metabolism pharmacology therapeutic use MeSH
- Butyrylcholinesterase metabolism MeSH
- Cholinesterase Inhibitors chemistry metabolism pharmacology therapeutic use MeSH
- Molecular Targeted Therapy * MeSH
- Blood-Brain Barrier metabolism MeSH
- Liver drug effects MeSH
- Rats MeSH
- Humans MeSH
- Ligands MeSH
- Neuroprotective Agents chemistry pharmacology therapeutic use MeSH
- Peptide Fragments chemistry MeSH
- Protein Aggregates drug effects MeSH
- Drug Design * MeSH
- Resveratrol MeSH
- Stilbenes chemistry MeSH
- Tacrine chemistry metabolism pharmacology therapeutic use MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Acetylcholinesterase MeSH
- amyloid beta-protein (1-42) MeSH Browser
- Amyloid beta-Peptides MeSH
- Antioxidants MeSH
- Butyrylcholinesterase MeSH
- Cholinesterase Inhibitors MeSH
- Ligands MeSH
- Neuroprotective Agents MeSH
- Peptide Fragments MeSH
- Protein Aggregates MeSH
- Resveratrol MeSH
- Stilbenes MeSH
- Tacrine MeSH
The research of novel antimycobacterial drugs represents a cutting-edge topic. Thirty phenolic N-monosubstituted carbamates, derivatives of salicylanilides and 4-chlorophenol, were investigated against Mycobacterium tuberculosis H37Ra, H37Rv including multidrug- and extensively drug-resistant strains, Mycobacterium avium, Mycobacterium kansasii, Mycobacterium aurum and Mycobacterium smegmatis as representatives of nontuberculous mycobacteria (NTM) and for their cytotoxic and cytostatic properties in HepG2 cells. Since salicylanilides are multi-targeting compounds, we determined also inhibition of mycobacterial isocitrate lyase, an enzyme involved in the maintenance of persistent tuberculous infection. The minimum inhibitory concentrations were from ≤0.5 μM for both drug-susceptible and resistant M. tuberculosis and from ≤0.79 μM for NTM with no cross-resistance to established drugs. The presence of halogenated salicylanilide scaffold results into an improved activity. We have verified that isocitrate lyase is not a key target, presented carbamates showed only moderate inhibitory activity (up to 18% at a concentration of 10 μM). Most of the compounds showed no cytotoxicity for HepG2 cells and some of them were without cytostatic activity. Cytotoxicity-based selectivity indexes of several carbamates for M. tuberculosis, including resistant strains, were higher than 125, thus favouring some derivatives as promising features for future development.
- Keywords
- Antimycobacterial activity, Carbamate, Cytotoxicity, Multi-targeting, Mycobacterium tuberculosis, Salicylanilide,
- MeSH
- Antitubercular Agents chemical synthesis chemistry pharmacology MeSH
- Hep G2 Cells MeSH
- Phenols chemical synthesis chemistry pharmacology MeSH
- Isocitrate Lyase antagonists & inhibitors metabolism MeSH
- Carbamates chemical synthesis chemistry pharmacology MeSH
- Humans MeSH
- Mycobacterium tuberculosis drug effects enzymology MeSH
- Salicylanilides chemical synthesis chemistry pharmacology MeSH
- Tuberculosis drug therapy MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Antitubercular Agents MeSH
- Phenols MeSH
- Isocitrate Lyase MeSH
- Carbamates MeSH
- salicylanilide MeSH Browser
- Salicylanilides MeSH
