Inactive Metabolite Research Articles

Discovery of benzyloxyphenyl- and phenethylphenyl-imidazole derivatives as a new class of ante–drug type boosters

Although cytochrome P450 3A4 (CYP3A4) inhibitors are used as boosters to increase drug absorption, the inhibition of CYP3A4 activity may affect the metabolism of other co-administered drugs. Therefore, we screened for and developed a new class of boosters to improve the oral availability of drugs. We identified benzyloxyphenyl imidazole and phenethylphenyl imidazole derivatives as new types of CYP3A4 inhibitors. Among the compounds synthesized, an ester 5c was found to inhibit CYP activity and the compound 5c was gradually converted to an inactive metabolite 5d under physiological conditions, indicating that the ester 5c may represent a novel ante-drug type booster.

In-depth investigation of the Silymarin effect on the pharmacokinetic parameters of sofosbuvir, GS-331007 and ledipasvir in rat plasma using LC-MS.

The use of complementary medicine (CMD) for liver support in Hepatitis C virus (HCV) patients sometimes coincides with the administration of oral antiviral drugs to eradicate the virus. This calls for a deep investigation of CMD effects on the pharmacokinetic parameters of these drugs to ensure their safety and efficacy. Silymarin (SLY), as a CMD, was selected to be given orally to healthy male rats with sofosbuvir (SFB) and ledipasvir (LED), a common regimen in HCV treatment. A new and sensitive LC-MS method was validated for the bioassay of SLY, LED, SFB and its inactive metabolite, GS-331007, in spiked plasma with lower limits of quantitation of 10, 1, 4 and 10 ng/ml, respectively. Moreover, the method was further applied to conduct a full pharmacokinetic profile of SFB, GS-331007 and ledipasvir with and without SLY. It was found that co-administration of SLY may expose the patient to unplanned high serum concentrations of SFB and LED. This could be accompanied by a decrease in SFB efficacy, potentially leading to therapeutic failure and the emergence of viral resistance.

Evaluation of UGT1A1 and CYP3A Genotyping and Single-Point Irinotecan and Metabolite Concentrations as Predictors of the Occurrence of Adverse Events in Cancer Treatment.

The variability on irinotecan (IRI) pharmacokinetics and toxicity has been attributed mostly to genetic variations in the UGT1A1 gene, responsible for conjugation of the active metabolite SN-38. Also, CYP3A mediates the formation of inactive oxidative metabolites of IRI. The association between the occurrence of severe adverse events, pharmacokinetics parameters, and UGT1A1 and CYP3A4 predicted phenotypes was evaluated, as the evaluation of [SN-38]/IRI dose ratio as predictor of severe adverse events. Forty-one patients undergoing IRI therapy were enrolled in the study. Blood samples were collected 15min after the end of drug the infusion, for IRI, SN-38, SN-38G, bilirubin concentrations measurements, and UGT1A1 and CYP3A genotype estimation. Data on adverse event was reported. Fifteen patients (36.5%) developed grade 3/4 adverse events. A total of 9.8% (n = 4) of the patients had UGT1A1 reduced activity phenotype, and 48.7% (n = 20) had UGT1A1 and 63.4% (n = 26) CYP3A intermediary phenotypes. Severe neutropenia and diarrhea were more prevalent in patients with reduced UGT1A1 in comparison with functional metabolism (50% and 75% versus 0% and 13%, respectively). SN-38 levels and its concentrations adjusted by IRI dose were significantly correlated to toxicity (rs = 0.31 (p = 0.05) and rs = 0.425 (p < 0.01)). The [SN-38]/IRI dose ratio had a ROC curve of 0.823 (95% CI 0.69-0.956) to detect any severe adverse event and 0.833 (95% CI 0.694-0.973) to detect severe diarrhea. The cut-off of 0.075ngmL-1mg-1 had 100% sensitivity and 65.7% specificity to predict severe diarrhea. Our data confirmed the relevance of the pre-emptive genotypic information of UGT1A1. The [SN-38]/IRI ratio, measured 15min after the end of the IRI infusion, was a strong predictor of severe toxicity and could be applied to minimize the burden of patients after IRI administration.

Zhx2 Is a Candidate Gene Underlying Oxymorphone Metabolite Brain Concentration Associated with State-Dependent Oxycodone Reward.

Understanding the pharmacogenomics of opioid metabolism and behavior is vital to therapeutic success, as mutations can dramatically alter therapeutic efficacy and addiction liability. We found robust, sex-dependent BALB/c substrain differences in oxycodone behaviors and whole brain concentration of oxycodone metabolites. BALB/cJ females showed robust state-dependent oxycodone reward learning as measured via conditioned place preference when compared with the closely related BALB/cByJ substrain. Accordingly, BALB/cJ females also showed a robust increase in brain concentration of the inactive metabolite noroxycodone and the active metabolite oxymorphone compared with BALB/cByJ mice. Oxymorphone is a highly potent, full agonist at the mu opioid receptor that could enhance drug-induced interoception and state-dependent oxycodone reward learning. Quantitative trait locus (QTL) mapping in a BALB/c F2 reduced complexity cross revealed one major QTL on chromosome 15 underlying brain oxymorphone concentration that explained 32% of the female variance. BALB/cJ and BALB/cByJ differ by fewer than 10,000 variants, which can greatly facilitate candidate gene/variant identification. Hippocampal and striatal cis-expression QTL (eQTL) and exon-level eQTL analysis identified Zhx2, a candidate gene coding for a transcriptional repressor with a private BALB/cJ retroviral insertion that reduces Zhx2 expression and sex-dependent dysregulation of cytochrome P450 enzymes. Whole brain proteomics corroborated the Zhx2 eQTL and identified upregulated CYP2D11 that could increase brain oxymorphone in BALB/cJ females. To summarize, Zhx2 is a highly promising candidate gene underlying brain oxycodone metabolite levels. Future studies will validate Zhx2 and its site of action using reciprocal gene editing and tissue-specific viral manipulations in BALB/c substrains. SIGNIFICANCE STATEMENT: Our findings show that genetic variation can result in sex-specific alterations in whole brain concentration of a bioactive opioid metabolite after oxycodone administration, reinforcing the need for sex as a biological factor in pharmacogenomic studies. The cooccurrence of female-specific increased oxymorphone and state-dependent reward learning suggests that this minor yet potent and efficacious metabolite of oxycodone could increase opioid interoception and drug-cue associative learning of opioid reward, which has implications for cue-induced relapse of drug-seeking behavior and for precision pharmacogenetics.

Differential Changes in Circulating Steroid Hormones in Hibernating Brown Bears: Preliminary Conclusions and Caveats.

Brown bears are obese when they enter the den, and after 6 mo of hibernation and physical inactivity, bears show none of the adverse consequences of a sedentary lifestyle in humans, such as cardiovascular disease, type 2 diabetes, and kidney failure. The metabolic mechanisms that drive hibernation physiology in bears are poorly defined, but systemic endocrine regulators are likely involved. To investigate the potential role of steroid hormones, we quantified the total levels of 12 steroid hormones, the precursor cholesterol, sex hormone-binding globulin (SHBG), and corticosterone-binding globulin (CBG) in paired serum samples from subadult free-ranging Scandinavian brown bears during the active and hibernation states. During hibernation, androstenedione and testosterone were significantly decreased in subadult female bears (), whereas they increased in all males but one () and therefore did not reach a significant difference. Despite this difference, SHBG increased more than 20-fold during hibernation for all bears. Compared with SHBG concentrations in humans, bear levels were very low in the active state, but during hibernation, levels equaled high levels in humans. The increased SHBG levels likely maintain a state of relative quiescence of the reproductive hormones in hibernating bears. Interestingly, the combination of SHBG and testosterone levels results in similar free bioavailable testosterone levels of 70-80 pM in both subadult and adult sexually active male bears, suggesting a role for SHBG in controlling androgen action during hibernation in males. Dehydroepiandrosterone sulfate, dihydrotestosterone, and estradiol levels were below the detection limit in all but one animal. The metabolically active glucocorticoids were significantly higher in both sexes during hibernation, whereas the inactive metabolite cortisone was reduced and CBG was low approaching the detection limit. A potential caveat is that the glucocorticoid levels might be affected by the ketamine applied in the anesthetic mixture for hibernating bears. However, increased hibernating cortisol levels have consistently been reported in both black bears and brown bears. Thus, we suggest that high glucocorticoid activity may support the hibernation state, likely serving to promote lipolysis and gluconeogenesis while limiting tissue glucose uptake to maintain a continuous glucose supply to the brain.

Review of remimazolam and sedatives in the intensive care unit.

Remimazolam is a novel intravenous ultra-short acting benzodiazepine that has the potential of being a safe and effective new sedative for use in intensive care unit (ICU) settings. Because remimazolam metabolizes rapidly by being hydrolyzed to an inactive metabolite (CNS 7054) through non-specific tissue esterase activity, specific dosing adjustment for older adults and for patients with renal or hepatic impairment patients (except for those with severe hepatic impairment) is not required. In addition, research has shown that remimazolam may be reversed by administration of flumazenil, as its half time was sufficiently short compared to flumazenil. It shows a lower incidence of cardiorespiratory depression, less injection pain, and no fatal complications such as propofol infusion syndrome and malignant hyperthermia of inhalational anesthetics. Future studies to study the suitability of remimazolam for managing the sedation of ICU patients who need sedation for a long time over several days is required.

3D-QSAR and scaffold hopping based designing of benzo[d]ox-azol-2(3H)-one and 2-oxazolo[4,5-b]pyridin-2(3H)-one derivatives as selective aldehyde dehydrogenase 1A1 inhibitors: Synthesis and biological evaluation.

Aldehyde dehydrogenase 1 (ALDH1A1), an oxidoreductase class of enzymes, is overexpressed in various types of cancer cell lines and is the major cause of resistance to the Food and Drug Administration (FDA)-approved drug, cyclophosphamide (CP). In cancer conditions, CP undergoes a sequence of biotransformations to form an active metabolite, aldophosphamide, which further biotransforms to its putative cytotoxic metabolite, phosphoramide mustard. However, in resistant cancer conditions, aldophosphamide is converted into its inactive metabolite, carboxyphosphamide, via oxidation with ALDH1A1. Herein, to address the issue of ALDH1A1 mediated CP resistance, we report a series of benzo[d]oxazol-2(3H)-one and 2-oxazolo[4,5-b]pyridin-2(3H)-one derivatives as selective ALDH1A1 inhibitors. These inhibitors were designed using a validated 3D-quantitative structure activity relationship (3D-QSAR) model coupled with scaffold hopping. The 3D-QSAR model was developed using reported indole-2,3-diones based ALDH1A1 inhibitors, which provided field points in terms of electrostatic, van der Waals and hydrophobic potentials required for selectively inhibiting ALDH1A1. The most selective indole-2,3-diones-based compound, that is, cmp 3, was further considered for scaffold hopping. Two top-ranked bioisosteres, that is, benzo[d]oxazol-2(3H)-one and 2-oxazolo[4,5-b]pyridin-2(3H)-one, were selected for designing new inhibitors by considering the field pattern of 3D-QSAR. All designed molecules were mapped perfectly on the 3D-QSAR model and found to be predictive with good inhibitory potency (pIC50 range: 7.5-6.8). Molecular docking was carried out for each designed molecule to identify key interactions that are required for ALDH1A1 inhibition and to authenticate the 3D-QSAR result. The top five inhibitor-ALDH1A1 complexes were also submitted for molecular dynamics simulations to access their stability. In vitro enzyme assays of 21 compounds suggested that these compounds are selective towardALDH1A1 over the other two isoforms, that is,ALDH2 and ALDH3A1. All the compounds were found to be at least three and two times more selective towardALDH1A1 over ALDH2 and ALDH3A1, respectively. All the compounds showed an IC50 value in the range of 0.02-0.80 μM, which indicates the potential for these to be developed as adjuvant therapy for CP resistance.

MP03-03 TARGETING V1/CHASE TO OVERCOME GEMCITABINE RESISTANCE

You have accessJournal of UrologyCME1 May 2022MP03-03 TARGETING V1/CHASE TO OVERCOME GEMCITABINE RESISTANCE Semih Sarcan, Daley S. Morera, Marie C. Hupe, Karina Aguilar, Anuj Sharma, Sung Alexander, Joseph McDaniels, Huabin Zhu, Martin Hennig, Martha Terris, Axel S. Merseburger, and Vinata B. Lokeshwar Semih SarcanSemih Sarcan More articles by this author , Daley S. MoreraDaley S. Morera More articles by this author , Marie C. HupeMarie C. Hupe More articles by this author , Karina AguilarKarina Aguilar More articles by this author , Anuj SharmaAnuj Sharma More articles by this author , Sung AlexanderSung Alexander More articles by this author , Joseph McDanielsJoseph McDaniels More articles by this author , Huabin ZhuHuabin Zhu More articles by this author , Martin HennigMartin Hennig More articles by this author , Martha TerrisMartha Terris More articles by this author , Axel S. MerseburgerAxel S. Merseburger More articles by this author , and Vinata B. LokeshwarVinata B. Lokeshwar More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002515.03AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Precision-based treatment approaches could improve the prognosis of patients with bladder cancer (BC). We recently discovered a first-in-class human Chondroitinase (Chase) that degrades chondroitin sulfate. This Chase is a splice variant of HYAL4 (V1) and promotes a malignant phenotype and chemoresistance in BC cells. We evaluated V1/Chase expression and functions in BC and inhibitors to overcome V1/Chase-mediated Gemcitabine (Gem) resistance. METHODS: Cohort 1: 86 bladder tissues (normal (NBL)=34; tumor (TBL)=52); cohort 2: 40 cystectomy specimens from MIBC patients who received adjuvant Gemcitabine plus cisplatin (G+C) treatment. Cohort 3: 144 urine specimens (BCA=42, non-BCa=102). Gene expression was measured by q-PCR in all cohorts and urine was assayed for Chase activity. V1/Chase was stably expressed or silenced in normal urothelial and three BC cell lines. Transfectants were analyzed for sensitivity to Gemcitabine and Cisplatin. The mechanism of Gem resistance was evaluated in preclinical in vitro and in vivo models. RESULTS: In cohort 1, V1 mRNA levels were an independent predictor of metastasis and death due to BC (P=0.0008). In cohort 2, high V1 levels significantly predicted with G+C treatment failure (P <0.0001). In cohort 3, Chase levels were 13 to 75-fold elevated in low and high-grade BC patients’ urine compared to non-BC cases (92.7% sensitivity; 88.2% specificity). V1-expressing urothelial and BC cells were resistant to Gem but not to Cisplatin. V1 expression increased Gem metabolism and subsequent efflux of an inactive metabolite dFdU. V1 increased CD44 shedding the conditioned media. This triggered intracellular activation of the JAK2/STAT3 pathway, resulting in the upregulation of cytidine deaminase (CDA). CDA induces Gem metabolism and dFdU efflux. JAK2, STAT3 and CDA inhibitors, (some FDA-approved), synergistically re- sensitized V1-expressing cells to Gem. V1-induced muscle invasive tumors that were metastatic. While Gem inhibited BC xenograft growth, V1-expressing tumors were resistant. Combination of Gem with a CDA inhibitor (tetrahydrouridine) abrogated V1 tumor growth with minimal toxicity. CONCLUSIONS: V1 is a functional biomarker that drives muscle-invasion, metastasis and Gem resistance in BC. Inhibitors of the JAK2/STAT3/CDA pathway overcome V1- mediated Gem resistance, suggesting a precision-based treatment approach to improve treatment response. Source of Funding: 1R01CA227277-01A1 (VBL); Department of Defence-PRCRP (W81XWH1810277 (CA170470; PRCRP) © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e19 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Semih Sarcan More articles by this author Daley S. Morera More articles by this author Marie C. Hupe More articles by this author Karina Aguilar More articles by this author Anuj Sharma More articles by this author Sung Alexander More articles by this author Joseph McDaniels More articles by this author Huabin Zhu More articles by this author Martin Hennig More articles by this author Martha Terris More articles by this author Axel S. Merseburger More articles by this author Vinata B. Lokeshwar More articles by this author Expand All Advertisement PDF DownloadLoading ...

Inhibition of Morphine Metabolism by Major Cannabinoids and their Metabolites

Opioids including morphine are mu‐opioid receptor (MOR) agonists that suppress excitatory neurotransmitter release, hyperpolarize neuronal cells, and decrease overall excitability, resulting in the desired analgesic effect. As a result of their over‐prescription, the still‐growing opioid epidemic has devastated many communities, with many individuals often using them in combination with other drugs including cannabis. Morphine is primarily metabolized by the phase II enzyme UDP‐glucuronosyltransferase (UGT) 2B7 to an inactive metabolite, morphine‐3‐glucuronide (M3G), and an active metabolite, morphine‐6‐glucuronide (M6G), both of which are extensively excreted into the urine. Previous studies in our lab have shown that major cannabis constituents including Δ9‐tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol (CBN) inhibit several major UGT enzymes. To examine whether cannabinoids or their major metabolites inhibit morphine glucuronidation by UGT2B7, screening assays were performed in pooled human liver microsomes (HLM) with the major cannabinoids and THC metabolites, 11‐hydroxy‐Δ9‐tetrahydrocannabinol (11‐OH‐THC), 11‐nor‐9‐carboxy‐Δ9‐tetrahydrocannabinol (11‐COOH‐THC), and 11‐nor‐Δ9‐tetrahydrocannabinol‐carboxylic acid glucuronide (THC‐COO‐Gluc). IC50 values were then determined for those cannabinoids that exhibited at least 50% inhibition at 100 µM final concentration. THC, 11‐OH‐THC, THC‐COO‐Gluc, and CBD all exhibited greater than 50% inhibition of both M3G and M6G formation at either 10 or 100 µM. The IC50 values for THC, 11‐OH‐THC, THC‐COO‐Gluc, and CBD against M3G formation were determined to be 27 ± 9.97 µM, 5.97 ± 2.31 µM, 15.58 ± 12.28 µM, and 15.66 ± 19.98 µM, respectively. The IC50 values for THC, 11‐OH‐THC, THC‐COO‐Gluc, and CBD against M6G formation were 14.49 ± 4.03 µM, 6.26 ± 0.76 µM, 13.22 ± 5.28 µM, and 8.12 ± 9.64 µM, respectively. To confirm results from these inhibition screenings, M3G and M6G formation were further examined using microsomal fractions from HEK293 cells overexpressing recombinant UGT2B7. THC, 11‐OH‐THC, THC‐COO‐Gluc, and CBD all exhibited &gt;50% inhibition of both M3G and M6G formation at either 10 or 100 µM in the recombinant enzyme system. These data suggest that several cannabinoids and their metabolites inhibit morphine metabolism, and that the co‐use of these agents may lead to adverse drug events in humans.

Development of an Automated, High‐Throughput Assay to Detect Angiotensin AT2‐Receptor Agonistic Activity by Nitric Oxide Measurement In Vitro

BackgroundThe angiotensin AT2‐receptor (AT2R) is a key component within the protective arm of the renin‐angiotensin system inducing vasodilation by nitric oxide (NO) production. Currently, no high‐throughput assay is available to identify AT2R agonists in vitro, which could explain the low number of AT2R selective ligands in drug development programs.ObjectiveTo design and validate a high‐throughput method for detection of AT2R activation in vitro based on NO release.MethodsHuman aortic endothelial cells (HAEC) were seeded into 96‐well plates at a density of 5000 cells per well. 48 hours after seeding, cells were loaded for 30 minutes with the NO probe, DAF‐FM diacetate (5µM). Cells were stimulated for 15 minutes with the AT2R‐agonist C21 (1µM) or with classical NO‐releasing agents as positive controls: bradykinin (BK, 0.1µM), acetylcholine (ACh, 1µM) or Ang‐(1‐7) (MasR agonist, 0.1µM). Cells were fixed and fluorescence signals captured by fluorescence microscopy with automated, built‐in image analysis (ImageXpress Pico, Molecular Devices, USA). AT2R‐transfected (AT2R‐CHO) or non‐transfected (NT‐CHO) Chinese hamster ovary cells were cultured and loaded with DAF‐FM diacetate according to the same protocol as HAEC. Concentration‐response curves were obtained for C21, BK, Ang II and Ang‐(1‐5) (0.1nM to 10µM each) using the automated protocol for ImageXpress Pico described above on AT2R‐CHO. To evaluate whether effects of the compounds in AT2R‐CHO were AT2R specific, the response to these compounds at the highest concentration (10µM) was also tested in NT‐CHO.ResultsUsing primary HAEC we could show that the automated, high‐throughput workflow was able to detect NO release by AT2R agonists as efficiently as by classical NO‐releasing agents [C21: 16.16 ± 6.84%; Ang‐(1‐7): 25.45 ± 3.37%, BK: 19.30 ± 2.92%; ACh: 24.01 ± 4.24%; one‐way ANOVA followed by Dunnett’s multiple comparison test, p≤0.05 vs control, ≥3 independent experiments for all compounds). In AT2R‐CHO, we detected that both C21 and Ang II (AT2R agonists) increased NO release in a concentration‐dependent manner, while BK had no effect (AT2R‐CHO lacks BK receptors) at any tested concentration. Surprisingly Ang‐(1‐5), a degradation product of Ang II thought to be inactive, induced NO release in a concentration‐dependent manner in AT2R‐CHO. In NT‐CHO, none of the compounds stimulated NO release, indicating that the effects by C21, Ang II and Ang‐(1‐5) in AT2R‐CHO were AT2R specific.ConclusionMeasurement of NO release from AT2R‐CHO or HAEC by DAF‐FM diacetate in an automated way is suitable as high‐throughput assay for the identification of AT2R‐agonistic compounds in vitro. Application of the assay revealed that Ang‐(1‐5), which has been regarded as an inactive metabolite of Ang II, has AT2R agonistic properties.

Hypogonadal Hypertension In Male Sprague‐Dawley Rats Is Reversed By Testosterone and its Genomically Inactive Metabolite 5β‐Dihydrotestosterone

Testosterone (TES) and its metabolites are efficacious vasodilators acutely, both in vitro and in vivo; however, their long‐term effects on blood pressure (BP) are unclear. We previously reported that castration (CsX) produced hypertension (HT) in normal male rats, and long‐term TES replacement therapy normalized BP and reduced renal renin‐angiotensin system (RAS) expression. Thus, in the present study, the effects of TES and 5β‐dihydrotestosterone (5β‐DHT), a metabolite with potent and efficacious vasodilator effects but lacking classic androgenic activity, were studied in CsX male Sprague‐Dawley rats. Rats were CsX at 12‐13 weeks age and then studied for 15 weeks. Weekly systolic BP (by tail cuff) revealed a progressive rise in BP over 10 weeks after CsX (110 ± 1.6 vs141 ± 1.2 mmHg). During the next 5 weeks, CsX rats received either TES (CsX + TES‐enanthate, 1.75 mg/kg, 2X/week) or 5β‐DHT‐propionate (CsX + 5β‐DHT, 2.00 mg/kg, 2X/week). BP gradually and progressively declined to normal by 5 weeks in CsX + TES rats (113 ± 1.3). In contrast, BP in CsX + 5β‐DHT rats was biphasic: a dramatic, more rapid decline to normal BP after only one week treatment (111 ± 0.7), and a continued more gradual decline to a hypotensive BP by 5 weeks (91 ± 1.1). Seminal vesicle (SV) weight after 5 weeks of androgen therapy (bioassay for androgenic activity) was 0.3250 ± 0.0180 g/100 g body weight in CsX + TES and 0.0122 ± 0.0006 in CsX + 5β‐DHT. In comparison, SV weights were 0.0160 ± 0.0010 in CsX without TES and 0.2860 ± 0.0110 in intact male rats. These data suggest that:1)endogenous and exogenous TES exert anti‐HT effects in male SD rats; 2) the TES metabolite 5β‐DHT exerts substantially greater, more rapid anti‐HT effects which appear to involve nongenomic, androgen receptor‐independent actions; and 3) while TES effects appear to involve diuretic/natriuretic effects on kidney via inhibition of renal RAS expression (our previous studies), the biphasic anti‐HT effects of 5β‐DHT in the present study appear to involve a rapid vasodilatory effect as well as a slower diuretic/natriuretic effect similar to that of TES. @font‐face {font‐family:“Cambria Math”; panose‐1:2 4 5 3 5 4 6 3 2 4; mso‐font‐charset:0; mso‐generic‐font‐family:roman; mso‐font‐pitch:variable; mso‐font‐signature:‐536870145 1107305727 0 0 415 0;}@font‐face {font‐family:Times; panose‐1:0 0 5 0 0 0 0 2 0 0; mso‐font‐charset:0; mso‐generic‐font‐family:auto; mso‐font‐pitch:variable; mso‐font‐signature:‐536870145 1342185562 0 0 415 0;}p.MsoNormal, li.MsoNormal, div.MsoNormal {mso‐style‐unhide:no; mso‐style‐qformat:yes; mso‐style‐parent:“”; margin:0in; mso‐pagination:widow‐orphan; text‐autospace:none; font‐size:12.0pt; font‐family:Times; mso‐fareast‐font‐family:“Times New Roman”; mso‐bidi‐font‐family:“Times New Roman”;}.MsoChpDefault {mso‐style‐type:export‐only; mso‐default‐props:yes; font‐size:11.0pt; mso‐ansi‐font‐size:11.0pt; mso‐bidi‐font‐size:11.0pt; font‐family:“Calibri”,sans‐serif; mso‐ascii‐font‐family:Calibri; mso‐ascii‐theme‐font:minor‐latin; mso‐fareast‐font‐family:Calibri; mso‐fareast‐theme‐font:minor‐latin; mso‐hansi‐font‐family:Calibri; mso‐hansi‐theme‐font:minor‐latin; mso‐bidi‐font‐family:“Times New Roman”; mso‐bidi‐theme‐font:minor‐bidi;}.MsoPapDefault {mso‐style‐type:export‐only; margin‐bottom:10.0pt; line‐height:115%;}div.WordSection1 {page:WordSection1;}

An Enhanced LC-MS-MS Technique for Distinguishing Δ8- and Δ9-Tetrahydrocannabinol Isomers in Blood and Urine Specimens.

Among the abundance of cannabinoids identified in cannabis, the active parent drug, Δ9-tetrahydrocannabinol (Δ9-THC), and its oxidized metabolite, 11-nor-9-carboxy-Δ9-THC (Δ9-THCCOOH), are attractive analytical targets to detect cannabis use. More recently, confirmation of these analytes may be hindered by a related interfering compound. Forensic toxicology laboratories attribute this phenomenon to an increase in cases containing Δ8-tetrahydrocannabinol (Δ8-THC) and 11-nor-9-carboxy-Δ8-THC (Δ8-THCCOOH). It is technically challenging to chromatographically resolve and accurately quantify Δ8- and Δ9-THC and THCCOOH in toxicology specimens due to their structural resemblance. This study describes a validated method to resolve and quantify active Δ8-THC and Δ9-THC in blood while qualitatively confirming the inactive metabolites Δ8-THCCOOH and Δ9-THCCOOH in blood and urine. Analytes are extracted and concentrated by solid-phase extraction and analyzed by liquid chromatography--electrospray ionization tandem mass spectrometry, which is amenable to modern toxicology laboratory routine workflows. This procedure offers a clear solution to untangling mixtures of these isomers, particularly in cases where Δ8-THC and its metabolite are the sole or dominant form.

Improvement of Albendazole Bioavailability with Menbutone Administration in Sheep.

Simple SummaryAnthelmintic drugs are among those most widely used in veterinary practice. The development of resistance to these drugs is a widespread problem, especially in small ruminants, and represents a serious threat to animal production. Thus, new possibilities to use the available pharmacological groups in a more efficient way should be explored. The objective of this study was to assess the pharmacokinetic interaction between a benzimidazole (albendazole, ABZ) and a choleretic drug (menbutone, MEN) in sheep, and it may result in greater effectivity of this drug against nematode parasites. Plasma concentrations of ABZSO (ABZ active metabolite) were higher when ABZ was administered with MEN. The proposed interaction is a simple, safe, and inexpensive way of increasing the effectivity of this anthelmintic widely used in livestock.The pharmacokinetic interaction between a benzimidazole (albendazole, ABZ) and a choleretic drug (menbutone, MEN) was evaluated in sheep. The plasma disposition of albendazole sulfoxide (ABZSO, active metabolite) and albendazole sulfone (ABZSO2, inactive metabolite) was investigated following an oral administration of albendazole (ABZ) (5 mg/kg) alone or with menbutone (MEN) (intramuscular, 10 mg/kg). Blood samples were collected over 3 days post-treatment, and drug plasma concentrations were measured by high performance liquid chromatography (HPLC). ABZSO was measured from 0.5 to 48 h, and ABZSO2 from 2 to 60 h. No parent drug was detected at any sampling time. Mean maximum plasma concentration (Cmax) and the area under the plasma concentration-time curve (AUC) were 12.8% and 21.5% higher for ABZSO when ABZ and MEN were administered together, which indicates a significant increase in the amount absorbed. The rate of absorption was not modified, with similar values for the time to reach Cmax (tmax) (11.5 h with ABZ + MEN and 10.7 h with ABZ treatment), although no significant differences were observed for these latter pharmacokinetic parameters. Regarding ABZSO2, Cmax, AUC and tmax values were similar after both treatments (ABZ or ABZ + MEN). The results obtained indicate that co-administration of ABZ and MEN may be an interesting and practical option to increase the efficacy of this anthelmintic.

Development of a Population Pharmacokinetic Model for the Diroximel Fumarate Metabolites Monomethyl Fumarate and 2-Hydroxyethyl Succinimide Following Oral Administration of Diroximel Fumarate in Healthy Participants and Patients with Multiple Sclerosis.

IntroductionDiroximel fumarate (DRF) is a next-generation oral fumarate that is indicated in the USA for relapsing forms of multiple sclerosis (MS). A joint population pharmacokinetic model was developed for the major active metabolite (monomethyl fumarate, MMF) and the major inactive metabolite (2-hydroxyethyl succinimide, HES) of DRF.MethodsMMF and HES data were included from 341 healthy volunteers and 48 patients with MS across 11 phase I and III studies in which DRF was administered as single or multiple doses. Population modeling was performed with NONMEM version 7.3 with the first-order conditional estimation method.ResultsEstimated MMF clearance (CLMMF), volume of distribution, and absorption rate constant (Ka) were 13.5 L/h, 30.4 L, and 5.04 h−1, respectively. CLMMF and HES clearance (CLHES) increased with increasing body weight. CLHES decreased with decreasing renal function. CLMMF and CLHES were 28% and 12% lower in patients with MS than in healthy volunteers, respectively. Ka was reduced in the presence of low-, medium-, and high-fat meals by 37%, 51%, and 67%, respectively, for MMF; and by 34%, 49%, and 62%, respectively, for HES.ConclusionsAge, sex, race, and baseline liver function parameters such as total bilirubin, albumin, and aspartate aminotransferase were not considered to be significant predictors of MMF or HES disposition.Supplementary InformationThe online version contains supplementary material available at 10.1007/s40120-021-00316-6.

Method To Visualize the Intratumor Distribution and Impact of Gemcitabine in Pancreatic Ductal Adenocarcinoma by Multimodal Imaging.

Gemcitabine (dFdC) is a common treatment for pancreatic cancer; however, it is thought that treatment may fail because tumor stroma prevents drug distribution to tumor cells. Gemcitabine is a pro-drug with active metabolites generated intracellularly; therefore, visualizing the distribution of parent drug as well as its metabolites is important. A multimodal imaging approach was developed using spatially coregistered mass spectrometry imaging (MSI), imaging mass cytometry (IMC), multiplex immunofluorescence microscopy (mIF), and hematoxylin and eosin (H&E) staining to assess the local distribution and metabolism of gemcitabine in tumors from a genetically engineered mouse model of pancreatic cancer (KPC) allowing for comparisons between effects in the tumor tissue and its microenvironment. Mass spectrometry imaging (MSI) enabled the visualization of the distribution of gemcitabine (100 mg/kg), its phosphorylated metabolites dFdCMP, dFdCDP and dFdCTP, and the inactive metabolite dFdU. Distribution was compared to small-molecule ATR inhibitor AZD6738 (25 mg/kg), which was codosed. Gemcitabine metabolites showed heterogeneous distribution within the tumor, which was different from the parent compound. The highest abundance of dFdCMP, dFdCDP, and dFdCTP correlated with distribution of endogenous AMP, ADP, and ATP in viable tumor cell regions, showing that gemcitabine active metabolites are reaching the tumor cell compartment, while AZD6738 was located to nonviable tumor regions. The method revealed that the generation of active, phosphorylated dFdC metabolites as well as treatment-induced DNA damage primarily correlated with sites of high proliferation in KPC PDAC tumor tissue, rather than sites of high parent drug abundance.

Management of Psychiatric Disorders in Patients with Chronic Kidney Diseases.

Management of Psychiatric Disorders in Patients with Chronic Kidney Diseases.

The curious case of Neuropathic Pain and its management: An overview

Abstract Neuropathic pain is a condition that occurs as a result of neural system damage or lesions. It could be peripheral or central neuropathic pain, depending on the location of the damage. Diabetes, spinal cord injury, numerous infections, cancer, and autoimmune illnesses are all potential causes of this condition. According to estimates, chronic pain has a prevalence rate of roughly 3% -17% worldwide. In most cases, neuropathic pain is stated to be “idiopathic” in origin, which means that its specific etiology is unknown; hence, pharmaceutical care of this condition is limited to treating its symptoms. The medications used to treat neuropathic pain fall into three categories: tricyclic antidepressants, gabapentinoids, and serotonin-norepinephrine reuptake inhibitors (SNRIs). Anticonvulsants, opioids, and topical medications are examples of different types of medications. Strong opioids, neurotoxins, and surgical alternatives are also used in treatment. In general, pharmacotherapy is frequently accompanied with accompanied with high doses, which results in a number of side effects. These medications are typically delivered orally, and drug absorption in systemic circulation leads to dispersion throughout the body, resulting in high peripheral circulation and concomitant side effects. Enzymatic degradation reduces bioavailability, while hepatic metabolism converts medicines to inactive metabolites. In such circumstances, an adequate amount of drug is unable to reach the brain due to the blood brain barrier, which hinders drug molecule permeability.

Efektivitas Agen Pendeplesi GSH pada Sitotoksisitas Cisplatin terhadap Sel Kanker: Systematic Literature Review

Cisplatin is one of chemotherapy agent for long cancer, ovarium cancer, gastric cancer, breast cancer, head-neck cancer. However, in the fact, the role of cisplatin does not always provide an optimal effect because it often appears cancer cell resistance phenomenon to cisplatin. This resistance condition occurs partly due to the inactive metabolite cause of conjugation reaction between cisplatin and GSH in cancer cells. Therefore, gluthathione (GSH) has an important role in controlling cisplatin resistance. This study aims to analyze some combination of cisplatin and the depletion agent of gluthathione (GSH) as a support for cisplatin activity in several types of cancer cells within in vitro scope. This study is prepared using systematic literature review method. Library search were carried out on two accredited international journals databases, namely PubMed and Science Direct with interval years of publication in 2011-2020. From 10 selected journals, it was shown that the use of GSH depletion agents could enhance the cytotoxic effect of cisplatin. This was analyzed based on data of the number measured GSH cells and the number of living cells (% cell viability) which gave a significant decrease. The result of research are expected to be able to provide information for the development of therapeutic agents on cisplatin as chemotherapy.

Effects of CYP2C9*3 and *13 alleles on the pharmacokinetics and pharmacodynamics of glipizide in healthy Korean subjects

Glipizide is a second-generation sulfonylurea antidiabetic drug. It is principally metabolized to inactive metabolites by genetically polymorphic CYP2C9 enzyme. In this study, we investigated the effects of CYP2C9*3 and *13 variant alleles on the pharmacokinetics and pharmacodynamics of glipizide. Twenty-four healthyKorean volunteers (11 subjects with CYP2C9*1/*1, 8 subjects with CYP2C9*1/*3, and 5 subjects with CYP2C9*1/*13) were recruited for this study. They were administered a single oral dose of glipizide 5mg. The plasma concentration of glipizide was quantified for pharmacokinetic analysis and plasma glucose and insulin concentrations were measured as pharmacodynamic parameters. The results represented that CYP2C9*3 and *13 alleles significantly affected the pharmacokinetics of glipizide. In subjects with CYP2C9*1/*3 and CYP2C9*1/*13 genotypes, the mean AUC0-∞ were increased by 44.8% and 58.2%, respectively (both P < 0.001), compared to those of subjects with CYP2C9*1/*1 genotype, while effects of glipizide on plasma glucose and insulin levels were not significantly different between CYP2C9 genotype groups. In conclusion, individuals carrying thedefective CYP2C9*3 and CYP2C9*13 alleles have markedly elevated plasma concentrations of glipizide compared with CYP2C9*1/*1 wild-type.

Brunner syndrome associated MAOA mutations result in NMDAR hyperfunction and increased network activity in human dopaminergic neurons

Monoamine neurotransmitter abundance affects motor control, emotion, and cognitive function and is regulated by monoamine oxidases. Among these, Monoamine oxidase A (MAOA) catalyzes the degradation of dopamine, norepinephrine, and serotonin into their inactive metabolites. Loss-of-function mutations in the X-linked MAOA gene have been associated with Brunner syndrome, which is characterized by various forms of impulsivity, maladaptive externalizing behavior, and mild intellectual disability. Impaired MAOA activity in individuals with Brunner syndrome results in bioamine aberration, but it is currently unknown how this affects neuronal function, specifically in dopaminergic (DA) neurons. Here we generated human induced pluripotent stem cell (hiPSC)-derived DA neurons from three individuals with Brunner syndrome carrying different mutations and characterized neuronal properties at the single cell and neuronal network level in vitro. DA neurons of Brunner syndrome patients showed reduced synaptic density but exhibited hyperactive network activity. Intrinsic functional properties and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR)-mediated synaptic transmission were not affected in DA neurons of individuals with Brunner syndrome. Instead, we show that the neuronal network hyperactivity is mediated by upregulation of the GRIN2A and GRIN2B subunits of the N-methyl-d-aspartate receptor (NMDAR), resulting in increased NMDAR-mediated currents. By correcting a MAOA missense mutation with CRISPR/Cas9 genome editing we normalized GRIN2A and GRIN2B expression, NMDAR function and neuronal population activity to control levels. Our data suggest that MAOA mutations in Brunner syndrome increase the activity of dopaminergic neurons through upregulation of NMDAR function, which may contribute to the etiology of Brunner syndrome associated phenotypes.