The “one-size fits all” model that has been used for decades is now replaced by the concept of the “right dose of the right drug for the right patient” ...
Since the emergence of the severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) at the end of 2019, no vaccine has been approved to counter this infection and the available treatments are ...mainly directed against the immune pathology caused by the infection. The coronavirus disease 2019 (COVID‐19) is currently causing a worldwide pandemic, pointing the urgent need for effective treatment. In such emergency, drug repurposing presents the best option for a rapid antiviral response. We assess here the in vitro activity of nilotinib, imatinib and dasatinib, three Abl tyrosine kinase inhibitors, against SARS‐CoV‐2. Although the last two compounds do not show antiviral efficacy, we observe inhibition with nilotinib in Vero‐E6 cells and Calu‐3 cells with EC50s of 1.44 μM and 3.06 μM, respectively. These values are close to the mean peak concentration of nilotinib observed at steady state in serum, making this compound a potential candidate for treatment of COVID‐19 in vivo.
•The cocktail approach monitor several CYP isoform activities simultaneously.•In vitro cocktails including 3–10 substrates are reviewed.•Crucial steps to select substrate cocktail are ...discussed.•Substrates interactions, inhibition, induction or phenotyping studies are presented.•A particular emphasis on incubations with human liver microsomes was assessed.
An assessment of cytochrome P450 (CYP) enzyme activity is essential for characterizing the phase I metabolism of biological systems or to evaluate the inhibition/induction properties of xenobiotics. CYPs have generally been investigated individually by single probes, and metabolite formation has been monitored by liquid chromatography-mass spectrometry (LC–MS). To increase the throughput, many probes have been applied to assess multiple CYP activities simultaneously within a single experiment. This strategy is called the cocktail approach, and it has already been reviewed for in vivo applications, but never for in vitro ones. This review focuses for the first time on an in vitro cocktail approach, and it references the most notable articles on this topic. The advantages and limitations of applying cocktails for the in vitro activity assessment of major human CYPs, namely, CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and subfamily CYP3A, are discussed. This article considers the probe reaction selections for each CYP according to regulatory recommendations, probe metabolic properties (i.e., specificity and turnover), probe concentrations and analytical sensitivity, but it also highlights a challenge specific to cocktail design, which is probe-probe interaction. The last part of the review reports some methodologies for incubating these cocktails and discusses some important issues regarding the incubation time, enzyme concentrations and sample preparation.
Tolerance and hyperalgesia associated with chronic exposure to morphine are major limitations in the clinical management of chronic pain. At a cellular level, neuronal signaling can in part account ...for these undesired side effects, but unknown mechanisms mediated by central nervous system glial cells are likely also involved. Here we applied data‐independent acquisition mass spectrometry to perform a deep proteome and phosphoproteome analysis of how human astrocytes responds to opioid stimulation. We unveil time‐ and dose‐dependent effects induced by morphine and its major active metabolites morphine‐3‐glucuronide (M3G) and morphine‐6‐glucuronide that converging on activation of mitogen‐activated protein kinase and mammalian target of rapamycin signaling pathways. We also find that especially longer exposure to M3G leads to significant dysregulation of biological pathways linked to extracellular matrix organization, antigen presentation, cell adhesion, and glutamate homeostasis, which are crucial for neuron– and leukocyte–astrocyte interactions.
Morphine, morphine‐3‐glucuronide (M3G) and morphine‐6‐glucuronide (M6G) activate human astrocytes via mitogen‐activated protein kinase and mammalian target of rapamycin signaling pathways. Morphine but especially prolonged exposure to M3G perturbed cellular pathways in astrocytes, which among others, are important for their interaction with neurons and leukocytes.
Coronavirus disease 2019 (COVID‐19), caused by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection, is a severe acute respiratory syndrome with an underlying inflammatory state. We ...have previously demonstrated that acute inflammation modulates cytochromes P450 (CYPs) activity in an isoform‐specific manner. We therefore hypothesized that COVID‐19 might also impact CYP activity, and thus aimed to evaluate the impact of acute inflammation in the context of SARS‐CoV‐2 infection on the six main human CYPs activity. This prospective observational study was conducted in 28 patients hospitalized at the Geneva University Hospitals (Switzerland) with a diagnosis of moderate to severe COVID‐19. They received the Geneva phenotyping cocktail orally during the first 72 hours of hospitalization and after 3 months. Capillary blood samples were collected 2 hours after cocktail administration to assess the metabolic ratios (MRs) of CYP1A2, 2B6, 2C9, 2C19, 2D6, and 3A. C‐reactive protein (CRP), interleukin 6 (IL‐6), and tumor necrosis factor‐α (TNF‐α) levels were also measured in blood. CYP1A2, CYP2C19, and CYP3A MRs decreased by 52.6% (P = 0.0001), 74.7% (P = 0.0006), and 22.8% (P = 0.045), respectively, in patients with COVID‐19. CYP2B6 and CYP2C9 MRs increased by 101.1% (P = 0.009) and 55.8% (P = 0.0006), respectively. CYP2D6 MR variation did not reach statistical significance (P = 0.072). As expected, COVID‐19 was a good acute inflammation model as mean serum levels of CRP, IL‐6, and TNF‐α were significantly (P < 0.001) higher during SARS‐CoV‐2 infection. CYP activity are modulated in an isoform‐specific manner by SARS‐CoV‐2 infection. The pharmacokinetics of CYP substrates, whether used to treat the disease or as the usual treatment of patients, could be therefore clinically impacted.
Cytochrome P450 (CYP) 2D6 metabolizes a wide range of xenobiotics and is characterized by a huge interindividual variability. A recent clinical study highlighted differential magnitude of CYP ...inhibition as a function of CYP2D6 genotype. The aim of this study was to investigate the effect of CYP2D6 genotype on the inhibition of dextromethorphan O‐demethylation by duloxetine and paroxetine in human liver microsomes (HLMs). The study focused on genotypes defined by the combination of two fully functional alleles (activity score 2, AS 2, n = 6), of one fully functional and one reduced allele (activity score 1.5, AS 1.5, n = 4) and of one fully functional and one non‐functional allele (activity score 1, AS 1, n = 6), which all predict extensive metabolizer phenotype. Kinetic experiments showed that maximal reaction velocity was affected by CYP2D6 genotype, with a decrease in 33% of Vmax in AS 1 HLMs compared to AS 2 (P = 0.06). No difference in inhibition parameters Ki, KI and kinact was observed neither with the competitive inhibitor duloxetine nor with the time‐dependent inhibitor paroxetine. Among the genotypes tested, we found no difference in absolute CYP2D6 microsomal levels with ELISA immunoquantification. Therefore, our results suggest that genotype‐sensitive magnitude of drug‐drug interactions recently observed in vivo is likely to be due to differential amounts of functional enzymes at the microsomal level rather than to a difference in inhibition potencies across genotypes, which motivates for further quantitative proteomic investigations of functional and variant CYP2D6 alleles.
Model-informed precision dosing (MIPD) has become synonymous with modern approaches for individualizing drug therapy, in which the characteristics of each patient are considered as opposed to ...applying a one-size-fits-all alternative. This review provides a brief account of the current knowledge, practices, and opinions on MIPD while defining an achievable vision for MIPD in clinical care based on available evidence. We begin with a historical perspective on variability in dose requirements and then discuss technical aspects of MIPD, including the need for clinical decision support tools, practical validation, and implementation of MIPD in health care. We also discuss novel ways to characterize patient variability beyond the common perceptions of genetic control. Finally, we address current debates on MIPD from the perspectives of the new drug development, health economics, and drug regulations.
Background
Agricultural by-products rich in lutein such as pistachio hull can be applied in pharmaceutical, cosmetics and food manufacturing. The development of rapid and cost-effective extraction ...methods of lutein from pistachio hull to optimize lutein recovery is of great interest to transpose to an industrial scale. Herein, we optimized the extraction protocol of lutein from the Iranian pistachio hull using experimental design and ultrasonic method.
Methods
Fresh raw un-hulled pistachios were harvested and dehulled, then hulls were dried and finely powdered to use for further analysis. Soxhlet process was carried out to obtain pistachio hull oleoresin and response surface methodology (RSM) was used for the optimization of saponification and ultrasonic methods. The lutein contents were quantitatively analyzed and validated using LC-MS/MS system.
Results
Our results showed that lutein in pistachio hull is mainly in free form, therefore the saponification method is not necessary for its extraction. Under optimal experimental design conditions, the maximum amount of lutein predicted and observed was 7.90 and 7.97 mg/100 g, respectively. Ethyl acetate was applied as an extraction solvent with the ultrasonic method followed by the setting up of the extraction time, temperature and solvent/sample ratio as variables. Under optimal experimental conditions corresponding to 45 min extraction time at 50 °C and 35.5 mg/ml of the solvent/sample ratio, the amount of lutein obtained from dried pistachio hull was 5.14 mg/100 g.
Conclusion
Pistachio waste products are rich in lutein which is in free from, so the administration of ultrasonic extraction using Ethyl acetate as a green and cost-effective method can be applied for lutein extraction from other plant materials and suggested for application on an industrial scale.
Graphical Abstract
Background
Measurement of neonatal renal function is challenging, and accurate, easy-to-use markers to estimate glomerular filtration rate (eGFR) are lacking. This study aimed to evaluate principal ...determinants of GFR in neonates and develop a predictive equation.
Methods
GFR was measured, using single injection inulin clearance, at median day 3 of life in 48 newborns. Associations of clearance with height, gestational age, weight, creatinine, and cystatin C were explored and a multivariable model to estimate GFR developed. We also evaluated preexisting GFR equations (Schwartz, Zappitelli, combined Zappitelli).
Results
Forty-four clearances were measured, 36 very preterm neonates (28–32 weeks); 5 extremely preterm (< 28 weeks), and 3 term newborns. No patient presented acute renal insufficiency. Median inulin clearance in preterm infants was 18.83 ml/min/1.73 m
2
(IQ 15.29; 24.99). Inulin clearance correlated with weight (
ρ
0.74), gestational age (
ρ
0.72), height (
ρ
0.49), and creatinine (
ρ
− 0.42), but not cystatin C. In the multivariable model, predicted GFR equation was 2.32* (weight (g))
0.64
/(creatinine (mcmol/l))
0.62
. Mean error in predicting clearance was − 0.8 ml/min/1.73 m
2
(− 3.0–1.4) ranging from − 14.9 to 13.3 ml/min/1.73 m
2
. Mean prediction error with Zappitelli and combined Zappitelli equations were 28.5 ml/min/1.73 m
2
(95% CI 24.6–32.3) and 28.3 ml/min/1.73 m
2
(95% CI 24.9–31.7), respectively, and 2 ml/min/1.73 m
2
(95% CI − 0.6–4.6) for Schwartz equation.
Conclusions
Weight and gestational age are crucial determinants of GFR in neonates. The Zappitelli models were not validated in our population. Our predictive model and Schwartz models performed better. Our model should be evaluated in another preterm population, particularly in those presenting renal insufficiency.
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