High Clearance Research Articles

Ustekinumab Drug Clearance Is Better Associated with Disease Control than Serum Trough Concentrations in a Prospective Cohort of Inflammatory Bowel Disease

Background/Objectives: This study aimed to compare the association of ustekinumab (UST) drug clearance (CL) and trough drug concentrations with disease activity in patients with inflammatory bowel diseases (IBDs). Methods: A prospective cohort of 83 patients with IBD receiving maintenance therapy with 90 mg subcutaneous UST was analyzed using Bayesian PK modeling. UST concentrations and antibodies to UST (ATU) were collected at the trough and measured using a drug-tolerant homogenous mobility shift assay (HMSA). CL was estimated using Bayesian estimation methods with priors from a previous population pharmacokinetic study specifically reparametrized using HMSA. Outcomes were combined clinical and biochemical remission and endoscopic healing index (EHI) score, a validated marker of endoscopic active disease in IBD. Statistical analysis consisted of linear and nonlinear mixed effect models for repeated time-to-event analysis. Results: A total of 83 patients with IBD were enrolled (median age 42 years, 52% female) and evaluated across 312 dose cycles (median follow-up: 279 days, median of 3 cycles/patient). Median concentrations and CL were 5.0 µg/mL and 0.157 L/day, respectively. Most patients (89%) were exposed to other biologics before starting UST, which was associated with lower rates of clinical and biochemical remission (p = 0.01). Longitudinal changes in concentrations were not associated with remission (p = 0.53). Conversely, higher CL was associated with a lower likelihood of remission (p < 0.01). EHI > 50 points (endoscopic active disease, n = 303 cycles) was associated with higher UST CL (p < 0.01). Conclusions: UST CL was more strongly associated with clinical and biochemical outcomes than trough concentrations, highlighting its potential role in therapy optimization.

A bioadhesive antioxidase-overexpressed probiotic prevents radiation enteritis by scavenging the excess reactive oxygen species

The scavenging of the excess reactive oxygen species (ROS) induced by radiation is fundamental for radiation protection. However, directly applying antioxidants results in low bioavailability and side effects. Superoxide dismutase (SOD) and catalase (CAT) have high ROS clearance efficiency, whereas their application is limited by the enzyme inactivation, making it difficult to exhibit significant therapeutic effects. Here, we engineered a probiotic Escherichia coli Nissle 1917 (EcN), i.e., AAEcN, serving as a SOD/CAT vehicle to scavenge ROS for the prevention and treatment of radiation enteritis (RE). The overexpressed Drsod and katE in AAEcN showed 5-fold ROS elimination efficiency compared to the wild EcN. Furthermore, the intestinal retention time of engineered EcN was prolonged through trefoil factor 3 gene (TFF3) modification of curli fibers on the bacterial surface, which contributed to the persistence of antioxidant enzyme activity. We found that AAEcN rapidly eliminated the intracellular ROS induced by radiation. Only a single oral dosing of AAEcN was satisfied to alleviate the radiation damage to the small intestine, colon, and spleen. Moreover, the homeostasis of pro-/anti-inflammatory cytokines was realized. The proliferation of the intestinal stem cells and spleen hematopoietic stem cells was enhanced, while the apoptosis of mucosal cells was inhibited. Our findings suggest valuable insights into the ROS scavenging way in RE, and establish an empirical basis for developing probiotics as an antioxidant enzyme vehicle for the bacteriotherapy of RE.

In vitro hydrolysis of areca nut xenobiotics in human liver

Areca nut (AN) is a substance of abuse consumed by millions worldwide, in spite of established oral and systemic toxicities associated with its use. Previous research demonstrates methyl ester alkaloids in the AN, such as arecoline and guvacoline, exhibit mood-altering and toxicological effects. Nonetheless, their metabolism has not been fully elucidated in humans. In the present study, an HPLC-UV bioanalytical method was developed to evaluate the hydrolytic kinetics and clearance rates of arecoline and guvacoline in human liver microsomes (HLM) and cytosol (HLC). The bioassay was capable of quantifying arecoline and guvacoline (and carboxylate metabolites arecaidine and guvacine, respectively) with good sensitivity, accuracy, and precision. Kinetics of arecoline and guvacoline hydrolysis best followed the Michaelis-Menten model. Apparent intrinsic clearance (Clint.in vivo) of arecoline was 57.8ml/min/kg in HLM and 11.6mL/min/kg in HLC, a 5-fold difference. Unexpectedly, guvacoline was dramatically less hydrolyzed than arecoline in both HLM and HLC, with Clint.in vivo estimates of 0.654ml/min/kg and 0.466ml/min/kg, respectively. These results demonstrate, for the first time, arecoline undergoes significant hydrolysis with high clearance rates in the liver. Furthermore, differential tissue metabolic rates and utilization of specific esterase inhibitors unequivocally demonstrated arecoline is a substrate for CES1 and not CES2.

In silico docking and ADMET evaluation of bioactive compounds from Phyllanthus niruri and captopril as angiotensin-converting enzyme (ACE) inhibitors for hypertension management

This study explores the molecular docking and ADMET properties of bioactive compounds from Phyllanthus niruri and captopril in relation to ACE inhibition for managing hypertension. The five natural compounds, including rutin, gallocatechin, quercitrin, astragalin, and kaempferol, were evaluated for their binding affinities through docking analysis. The results demonstrated that these compounds exhibit strong binding energies, with rutin showing the highest affinity (-9.8 kcal/mol) compared to captopril (-5.5 kcal/mol). ADMET profiling revealed that the natural compounds, particularly rutin and kaempferol, possess favorable pharmacokinetic properties, including high GI absorption and low toxicity. Additionally, while captopril exhibited high protein binding and moderate clearance, the natural compounds presented lower toxicity across various parameters, such as hepatotoxicity and genotoxicity. Overall, these findings suggest that Phyllanthus niruri compounds could serve as promising natural alternatives to captopril for ACE inhibition, offering an improved safety profile and favorable drug-likeness properties.

Two Demosponges as Promising Bioremediators of a Potential Pathogenic Vibrio

Marine sponges play a fundamental role in the proper functioning of the ecosystem by filtering organic matter and contributing to nutrient fluxes. These animals have been proposed as efficient bioremediators of microbiological contamination in various environmental conditions subjected by anthropogenic pressure. In the present study, the bioremediation potential of the demosponges Aplysina aerophoba and Geodia cydonium was analyzed ex situ. For this purpose, the viable count of an antibiotic-resistant bacterial strain belonging to the species Vibrio parahaemolyticus was assessed in presence of the selected sponge species. Although some sponge individuals showed closed oscula during the first hours of the experiment, A. aerophoba and G. cydonium reduced the bacterial load in the seawater up to five orders of magnitude in 72 h. In addition, they had high clearance rates and retention efficiencies, with almost complete removal of the tested bacteria. Low Vibrio concentrations were observed in all tanks after six days, suggesting no excretion of viable Vibrio from sponges. These results corroborate the usefulness of A. aerophoba and G. cydonium as bioremediators of bacteria and therefore appear to be ideal candidates for bioremediation purposes in anthropogenic environments, such as aquaculture facilities, where multidrug-resistant bacteria may play a role in the spread of antimicrobial resistance.

Properties of Uremic Solutes That Allow Their Effective Control by Hemodialysis.

If the GFR falls far enough, uremic symptoms such as anorexia and nausea prompt the initiation of dialysis. Thrice weekly hemodialysis can prevent recurrence of these symptoms even when patients become anuric. To accomplish this it must maintain the plasma levels of the uremic solutes which cause these symptoms lower than they were when dialysis was initiated. This study examined kinetic properties that solutes must possess for hemodialysis to accomplish this. We also sought to identify uremic solutes that possess these properties. Mathematical modeling analyzed how a solute's kinetic properties would determine the relation of its level in an anuric dialysis patients to its level when uremic symptoms prompt dialysis initiation. The previously unstudied solute methylurea was assayed by liquid chromatography tandem mass spectrometry (LC/MS/MS) in 13 participants on hemodialysis, 9 participants with advanced CKD, and 10 participants without kidney disease. Mathematical modeling showed that conventional dialysis can effectively control the plasma levels better than the failing native kidneys only of solutes which have a high dialytic clearance relative to their native kidney clearance and a large volume of distribution. LC/MS/MS measurements showed that methylurea has these properties. The dialytic clearance of methylurea was 255 ± 32 ml/min and its volume of distribution was 1.09 ± 0.25 times the body water volume in hemodialysis patients. The methylurea clearance was lower than the GFR in patients without kidney disease (fractional clearance 0.44 ± 0.19) and patients with advanced CKD (fractional clearance 0.53 ± 0.10). Literature review revealed that urea was the only solute previously known to possess these properties. A further search for solutes whose properties include a high dialytic clearance, a relatively low native kidney clearance, and a high volume of distribution could help identify solutes that contribute to uremic symptoms.

DOP091 Early serum infliximab levels predict outcomes in Acute Severe Ulcerative Colitis: results from PREDICT-UC

Abstract Background The utility of infliximab (IFX) therapeutic drug monitoring (TDM) in acute severe ulcerative colitis (ASUC) is unclear. We aimed to assess whether IFX levels are associated with outcomes in ASUC. Methods PREDICT-UC (NCT02770040) was a randomised controlled trial that compared dosing strategies in 138 steroid-refractory ASUC patients.1 Serum and faecal IFX levels were quantified by ELISA (MabTrack level infliximab, Essange Reagents, Netherlands) after conclusion of the trial and correlated with outcomes: IFX response by day 7 (Lichtiger score [LS]<10, with ≥3-point reduction and decrease in rectal bleeding and stool frequency ≤4/day); eventual response by day 14 (LS<10); and colectomy by month 3. Individual IFX clearance was estimated using a two-compartment pharmacokinetic model with fixed V1, V2 and Q. Results 681 serum IFX levels were available across 135 patients; 91 received an initial 5mg/kg and 44 an initial 10mg/kg IFX dose. 85 responded by day 7 and 17 required colectomy by month 3. Post-IFX serum levels were higher on days 1 and 3 (median ug/mL, IQR) in the 10mg/kg group (175.4, 137.2-202.7 and 116.3, 83.4-132.9) compared to the 5mg/kg group (91.8, 77.2-109.4 and 56.0, 46.0-67.0; each P<0.001). Day 1 and day 3 serum IFX levels were not significantly different in responders and non-responders. A higher day 3:day 1 serum IFX ratio predicted response (63.1%, IQR 56.0-72.1 vs 58.1%, IQR 51.6-62.8, P=0.006; AUC 0.67). A lower day 3 serum IFX level predicted colectomy (51.0, IQR 39.2-57.4 vs 69.0, IQR 51.1-101.7, P=0.003; AUC 0.23). IFX clearance using serum levels between days 1-7 was higher in non-responders compared to responders (0.72L/day, IQR 0.55-0.89 vs 0.56L/day, IQR 0.39-0.72, P<0.001) and in patients who had colectomy (P=0.011). Patients with high clearance (≥0.62L/day) were more likely to respond to an initial 10mg/kg vs 5mg/kg IFX dose (RR 1.50, 95%CI 1.01-2.23, P=0.046), and more likely to require colectomy if they received an initial 5mg/kg vs 10mg/kg dose (HR 4.81, 95%CI 1.09-21.37, P=0.039). In patients with high clearance who did not respond initially, response by day 14 was higher in those receiving a second 10mg/kg dose compared to 5mg/kg (10/26 [38%] vs 1/9 [11%]; RR 3.43, 95%CI 1.05-11.19, P=0.041). Day 1 and 3 faecal IFX correlated with IFX clearance (both rho=0.36, both P<0.001), CRP and the UCEIS (including bleeding and erosion/ulcer sub-scores). Conclusion Elevated day 3:day 1 serum IFX ratio was associated with IFX response by day 7. Early IFX clearance predicted IFX response and month 3 colectomy. High IFX clearance may be overcome by higher IFX dosing, resulting in improved response and reduced colectomy rates. Early IFX level quantification can help predict outcomes in ASUC.

P0972 Higher vedolizumab clearance associates with poor therapeutic outcomes during maintenance therapy of vedolizumab in Crohn’s disease

Abstract Background Drug Clearance (CL) is a better pharmacokinetic (PK) metric compared to trough levels (TL) in associating with therapeutic outcome of infliximab, adalimumab and ustekinumab therapy in Crohn’s disease (CD). However, data regarding vedolizumab (VDZ) are limited. We compared the performances of VDZ CL and TL in associating with therapeutic outcomes in CD. Methods This retrospective analysis included a subset patients from the LOVE-CD trial at Amsterdam UMC. Blood samples were collected at trough during intravenous VDZ maintenance treatment and stored at -80ºC until analysis. VDZ TL were measured using homogenous mobility shift assay and CL was estimated using nonlinear mixed effect models. Therapeutic outcomes consisted of endoscopic remission (Simple Endoscopic Scoring for CD [SES-CD] <3) measured at one-year after starting treatment, and clinical and biochemical remission status (CRP <3 mg/L with CD activity index <150) at each maintenance cycle. Statistical analyses included receiver operating characteristics (ROC) curves, area under the curve (AUC) and logistic regression. Results A total of 50 patients (24 females, mean age 37 years) were evaluated in this analysis. At one year 39 patients had endoscopic data; 17/39 (44%) were in endoscopic remission. In the 50 patients, 38% of cycles (120/312, median 6 per patients) showed clinical and biochemical remission. Median TL and CL were 16.0 µg/mL (IQR: 9.5-23.8 µg/mL) and 0.154 L/day (IQR: 0.125-0.190 L/day) respectively. Anti-drug antibodies to VDZ were negligible in this cohort (2.5%, 8/323 specimens). ROC analysis yielded higher AUC with CL (AUC= 0.824 [95%CI: 0.683-0.964]) than with TL (AUC=0.606 [95%CI: 0.421- 0.790]) (difference= 0.218 95%CI: 0.074- 0.362; p=0.003) in distinguishing endoscopic remission from endoscopic active disease. Similar results were observed with the clinical and biochemical remission outcome (AUC [conc]=0.530 [95%CI: 0.466-0.595] compared to AUC[CL]=0.646 [95%CI: 0.584- 0.709]) (difference: 0.116 95%CI: 0.063- 0.169; p<0.001). Logistic regression revealed that TL were not significantly associated with either of the two outcomes (p>0.28) (Figure). In contrast, higher CL (>0.156 L/day) was associated with 9.0-fold (95%CI: 1.7,44.0) and 2.1-fold (95%CI: 1.3,3.4) lower likelihood of endoscopic and clinical & biochemical remission, respectively (p<0.01) (Table). Conclusion These data support the hypothesis that VDZ CL is better than TL in associating with outcomes of VDZ treatment in CD. Given that there was little immune response to VDZ in this study, the association between higher CL and poor outcome most likely reflects higher inflammatory burden that consumes the drug from the central compartment.

DOP063 Hospitalised severe ulcerative colitis patients treated with rescue infliximab risk treatment failure due to underexposure caused by high drug clearance

Abstract Background Acute severe ulcerative colitis (ASUC) occurs in 25% of patients with ulcerative colitis (UC) resulting in hospital admission and treatment with corticosteroids followed by rescue therapy with infliximab, if needed. High baseline infliximab clearance at admission has been associated with risk of colectomy supporting an intensified infliximab regimen in case of insufficient effect.1 However, a recent randomized controlled trial did not find superior outcomes of different intensified infliximab regimens.2 We investigated if admitted patients with severe UC have increased clearance of infliximab. Methods Infliximab concentrations from 154 UC patients were retrieved from our biobank resulting in 248 (81%) induction phase and 58 (19%) maintenance phase samples. These data were used to build a non-linear mixed effect model describing infliximab pharmacokinetics (PK) in UC. Variables tested for potentially influencing clearance comprised clinical disease activity (Mayo/SCCAI, 86% of assessments; physician’s global assessment, 14%), endoscopic activity (Mayo) (56%), CRP, albumin, patient demographics, and whether patients were treated as outpatients or hospitalised, including formal fulfilment of Truelove Witt’s criteria. Results A 2-compartment model with a linear clearance described infliximab PK, in line with previous reports.3 Hence, central and peripheral volumes of distribution were 6.52 L and 2.56 L, respectively, clearance 0.339 L/day and intercompartmental clearance 0.146 L/day. Hospitalisation status was the most impactful variable significantly influencing infliximab clearance in the model. Hence, hospitalised patients with severe UC (32% of study population) had a 35% [90%CI 14-56%] increased infliximab clearance compared to outpatients (median clearance of 0.463 L/day vs. 0.339 L/day, p<0.0001). This resulted in a substantial decreased IFX exposure from week 2 onwards. Thus, 74% of hospitalised patients had sub-therapeutic infliximab concentration at week 2 (<20 ug/mL4), 69% at week 6 (<15 ug/mL4), and 56% at week 14 (<7 ug/mL4). Formal fulfilment of Truelove Witt’s criteria did not result in additional increase in infliximab clearance among hospitalized patients (ASUC median clearance 0.442 L/day vs non-ASUC 0.516 L/day, p=0.08). Addition of albumin did not significantly improve the model. Conclusion Infliximab clearance is substantially increased among hospitalised patients with severe UC, resulting in underexposure in the majority of patients treated with a standard dosing regimen. An intensified regimen should be considered preferably supported by therapeutic drug monitoring and PK modelling.

Metabolically Stable Adenylation Inhibitors of Biotin Protein Ligase as Antibacterial Agents.

The antibacterial agent Bio-AMS is metabolized in vivo through hydrolysis of the central acyl-sulfamide linker leading to high clearance and release of a moderately cytotoxic metabolite M1. Herein, we disclose analogues designed to prevent the metabolism of the central acyl-sulfamide moiety through steric hindrance or attenuation of the acyl-sulfamide electrophilicity. Bio-9 was identified as a metabolically stable analogue with a single-digit nanomolar dissociation constant for biotin protein ligase (BPL) and minimum inhibitory concentrations (MICs) against Mycobacterium tuberculosis and Staphylococcus aureus ranging from 0.2 to 20 μM. The antibacterial activity of Bio-9 was dependent on BPL expression level and was more than 70-fold better against a strain underexpressing BPL and, conversely, more than 5-fold less effective against a strain overexpressing BPL. Pharmacokinetic and metabolic studies demonstrated that Bio-9 was metabolically stable in vivo, showing negligible hydrolysis that translated to substantially reduced clearance and concomitantly boosted drug exposure and half-life compared to Bio-AMS.

Population Pharmacokinetics of Sotorasib in Healthy Subjects and Advanced Solid Tumor Patients Harboring a KRASG12C Mutation from Phase 1 and Phase 2 Studies.

Sotorasib is a novel KRASG12C inhibitor that has shown robust efficacy, safety, and tolerability in patients with KRASG12C mutation. The objectives of the population pharmacokinetic (PK) analysis were to characterize sotorasib population PK in healthy subjects and patients with advanced solid tumors with KRASG12C mutation from 6 clinical studies, evaluate the effects of intrinsic and extrinsic factors on PK parameters, and perform simulations to further assess the impact of identified covariates on sotorasib exposures. A two-compartment disposition model with three transit compartments for absorption and time-dependent clearance and bioavailability well described sotorasib PK. Sotorasib exposure increased in a less-than-dose proportional manner across the evaluated 180mg to 960mg dose range. Disease burden, measured by Eastern Cooperative Oncology Group (ECOG) score and baseline tumor size, were identified as significant covariates on clearance. Lower disease burden was associated with higher clearance (and thus lower sotorasib exposure). Low albumin was found to be associated with lower clearance of sotorasib. Use of PPIs was associated with reduced sotorasib exposures. Although sex, race, and high fat meal were significantly associated with PK parameters, their impact on exposures were not clinically relevant. Other evaluated covariates, including body weight, age, renal impairment (evaluated by chronic kidney disease stage score) and hepatic impairment (evaluated by NCI criteria) were not statistically significant. Greater baseline disease burden and low albumin were associated with lower sotorasib clearance; based on the established efficacy and safety profiles from clinical trials, the estimated magnitude of these effects does not warrant a dose adjustment.

Improving the bioactivity of cellulose acetate hemodialysis membranes through nanosilver modification.

The effectiveness and safety of hemodialysis can be hindered by protein accumulation, mechanical instability of membranes and bacterial infection during the dialytic therapy. Herein, we show that cellulose acetate membranes modified with the low-fouling polymers (namely polyvinylpyrrolidone and polyethylene glycol), followed by the in situ reduction of different densities of silver oxide(I) nanoparticles, can effectively address these limitations. These improvements comprise the enhanced resistance to the protein fouling, improved antimicrobial capabilities against S. aureus, increased selectivity, and thermal stability and mechanical strength. The nano-enhanced membranes showed an improved albumin rejection rate of approximately 90%, and the creatinine clearance rate ranged between 90 and 94%. Our findings demonstrate that nanosilver-modified membranes can be readily prepared from precursor solutions to act as robust, biocompatible, and hydrophilic hemodialysis membranes with controlled bacteriostatic potential, antifouling properties and high toxin clearance.

Population pharmacokinetics of bedaquiline: a systematic review.

Bedaquiline (BDQ) plays a critical role in the treatment of multidrug-resistant tuberculosis (MDR-TB). However, the large pharmacokinetic (PK) variability of BDQ presents a significant challenge in its clinical use. This study aimed to summarize the population PK characteristics of BDQ and to identify significant covariates affecting the PK variation of BDQ. The PubMed and Web of Science databases were systematically searched from their inception to October 1, 2023. Population pharmacokinetics (PPK) studies on BDQ were searched and identified in this review. The PK characteristics of included studies and the significant covariates were systematically summarized. Eight studies conducted in adults and one in children and adolescents were included in this review. A three disposition compartments with dynamic absorption transport chamber model was the commonly used structural model for BDQ. Body weight, race, albumin, and concomitant medication were significant covariates affecting BDQ PK variation. With the increase of weight and albumin levels, the clearance (CL) of BDQ was gradually increased. The average CL value per body weight in children was 1.49-fold higher than that in adults. Black race patients had an 84% higher CL than other populations. Moreover, combined with rifampicin and rifapentine, BDQ had 378% and 296% higher clearance rates, respectively. Body weight, race, albumin level, and concomitant medication may be important factors affecting patients' exposure differences. Further PPK studies of BDQ are needed to facilitate optimal dosing regimens.

In silico studies on quercetin, myricetin, and kaempferol in inhibiting TGF-β1 and galectin-3 for cardiac fibrosis management

Cardiac fibrosis remains as the leading cause of death worldwide and is often associated with elevated levels of transforming growth factor-β 1 (TGF-β1) and galectin-3, making them potential therapeutic targets. Recent studies revealed that quercetin, myricetin, and kaempferol have the biological effect for several cardiovascular diseases. However, the investigation into this topic through molecular models and analysis remain unexplored. The aim of this study was to evaluate the potential effect of quercetin, myricetin, and kaempferol which targeted TGF-β1 and galectin-3. In this study, quercetin, myricetin, and kaempferol roled as the tested ligands. Subsequently, colchicine and native ligand acted as control ligands that were screened through molecular docking against TGF-β1 and galectin-3 using AutoDock tools to identify the potential inhibitor. The stability of ligand-receptor complexes was assessed through molecular dynamic (MD) simulations using NMAD. Absorption, Distribution, Metabolism, Excretion and toxicity (ADMET) prediction were also performed using ADMETlab 2.0. Molecular docking analysis revealed that quercetin, myricetin, and kaempferol exhibited strong binding affinity which are -8.9 kcal/mol, -8.5 kcal/mol, -7.6 kcal/mol respectively with TGF-β1, and -7.5 kcal/mol, -7.0 kcal/mol, -5.7 kcal/mol respectively with galetcin-3; low inhibition constant (Ki); and stable interaction with the active sites of TGF-β1 and galectin-3. MD simulations confirmed the stability and compactness of the ligand-receptor complexes. ADMET analysis also showed high Plasma Protein Binding (PPB) values (quercetin: 95%, myricetin: 92%, and kaempferol: 97%) and moderate clearance values (quercetin: 8.284%, myricetin, and 7.716%, kaempferol: 6.868%) for the tested compounds. In conclusion, the in silico analyses suggested that quercetin, myricetin, and kaempferol are promising for cardiac fibrosis therapies by inhibiting TGF-β1 and galectin-3.

Purification of AAV8 through a scalable two-step monolithic chromatography approach.

Adeno-associated viruses (AAV) are becoming increasingly popular as a powerful tool for gene delivery therapy applications. Although processes to produce AAV are established, future demand for this type of viral vector requires further development of manufacturing processes to make them more robust, scalable, and flexible to accommodate the rise of engineered capsids. This study focuses on designing and evaluating a two-step chromatography process for capturing and polishing AAV8 using monolith chromatography media. A cation-exchange-based capture step was established, using CIMmultus® SO3, resulting in a virus recovery of approximately 70 % of total capsids. This step also achieved high protein removal (>95 %) and considerable DNA clearance (>80 %). For the polishing step, three different CIMmultus® monoliths were evaluated: anion-exchange-based QA (strong anion exchange), multimodal-based PrimaS (weak anion exchange and hydrogen bonding) and multimodal-based PrimaT (weak anion exchange, hydrogen bonding and metal affinity) ligands. High viral vector genome recoveries, DNA and protein clearance and a three-fold full capsid enrichment were observed at a 1 mL column scale. Similar results were obtained during a scale-up to 8 mL and 4 mL monolith volumes for capture and polishing, respectively. These results provide a strong and robust alternative to conventional AAV purification methods, such as resin-based affinity chromatography and density gradient ultracentrifugation.

Novel tertiary diarylethylamines as functionally selective agonists of the kappa opioid receptor.

Novel kappa opioid receptor (KOR) agonists that preferentially activate G-protein signaling versus β-arrestin-2 recruitment are described. Starting from a literature-reported phenol-containing diphenethylamine KOR agonist, structure-activity relationship (SAR) studies revealed replacement of the phenol with various non-hydroxylated bicyclic heteroaromatics led to tertiary diarylethylamines which retained KOR agonist activity and improved metabolic stability in human liver microsomes. Further optimizations produced compound 39, a potent activator of G-protein signaling (GTPγS EC50 = 14 nM, 83 % Emax) that did not elicit a β-arrestin-2 recruitment functional response (Emax < 10 %). Compound 39 demonstrated moderate to high intrinsic clearance in human hepatocytes and low potential for Pgp-mediated efflux when evaluated in the MDR1-MDCK permeability assay. Compound 39 exhibited 60- and 810-fold selectivities versus the related mu (MOR) and delta (DOR) opioid receptors in recombinant radioligand binding (Ki) assays. These findings highlight compound 39 and related structures as potential leads toward safe and tolerable therapeutics that target central nervous system (CNS) disorders for which KOR agonism could provide benefit.

Development and First-in-Human evaluation of a Site-Specific [18F]-Labeled PD-L1 nanobody PET radiotracer for noninvasive imaging in NSCLC.

Immunohistochemistry (IHC) for PD-L1 detection is limited by its invasiveness and heterogeneity of tumors. To address these challenges, a new PD-L1-targeted nanobody-based immune-PET radiotracer [18F]AlF-APN09 was developed using the site-specific radiolabeling method with the complexing agent (Mal-RESCA) under mild conditions. [18F]AlF-APN09 was prepared at room temperature (pH 4.6-4.8) within 20min with satisfactory radiochemical yields (45.8±4.48%, non-decay corrected), high radiochemical purity (>98%) and moderate apparent molar activity (15-35GBq/μmol), and remained stable in both PBS and 5% HSA after 4h (>90%). Cell uptake studies indicated variable levels of surface PD-L1 expression in the following order: A549PD-L1>H1975>A549. In micro-PET/CT imaging, A549PD-L1 and H1975 tumors were distinctly visualized in a 6.0:1 and 3.2:1 ratios over PD-L1-negative A549 tumors in vivo. Ex vivo biodistribution studies showed tumor uptake values of 6.47±1.06%ID/g (A549PD-L1) and 2.27±0.19%ID/g (H1975), significantly higher than 0.90±0.28%ID/g in A549 tumors. The estimated effective radiation dose in humans was 8.65E-03mSv/MBq, lower than that of conventional [18F]FDG. First-in-human imaging was conducted on a single resectable non-small cell lung cancer (NSCLC) subject without any adverse reactions. The radiotracer exhibited renal excretion with minimal hepatobiliary clearance. Tumor uptake reached SUVmax 4.20 at 2h post-injection, demonstrating high contrast and rapid clearance. After PD-1 inhibitor immunotherapy and chemotherapy, the subject showed a therapeutic response and postoperative pathological specimens confirmed a major pathological response (MPR). These results suggest that we have successfully developed a new PD-L1-targeted nanobody PET tracer using the site-specific labeling method with the complexing agent (Mal-RESCA) within 20min under mild conditions and [18F]AlF-APN09 is a promising noninvasive PET radiotracer for visualizing PD-L1 expression in tumors, offering rapid tumor targeting, excellent signal-to-noise ratios, and favorable clearance properties.

A Self-Reinforced "Microglia Energy Modulator" for Synergistic Amyloid-β Clearance in Alzheimer's Disease Model.

Microglial phagocytosis is a highly energy-consuming process that plays critical roles in clearing neurotoxic amyloid-β (Aβ) in Alzheimer's disease (AD). However, microglial metabolism is defective overall in AD, thereby undermining microglial phagocytic functions. Herein, we repurpose the existing antineoplastic drug lonidamine (LND) conjugated with hollow mesoporous Prussian blue (HMPB) as a "microglial energy modulator" (termed LND@HMPB-T7) for safe and synergistic Aβ clearance. The modified blood-brain barrier penetrating heptapeptide (T7) enables efficient transport of LND@HMPB-T7 to the AD brain. LND in LND@HMPB-T7 could fuel Aβ phagocytosis by stimulating microglial adenosine triphosphate (ATP) production, whereas HMPB with catalase and superoxide dismutase-mimicking activities substantially alleviates the mitochondrial side effects commonly associated with LND and thus further enhances ATP production. The synergism of LND and nanozyme affords a high microglial Aβ clearance efficacy without triggering mitochondrial dysfunction. In vivo experiments ascertain that LND@HMPB-T7 could synergistically promote phagocytic clearance of Aβ, relieve neuroinflammation and ameliorate cognitive function in AD mice. These findings indicate that LND@HMPB-T7 holds tremendous clinical potential as a repurposed drug for AD treatment.

Fabrication and Performance Evaluation of a Novel Composite PVC-ZnO Membrane for Ciprofloxacin Removal by Polymer-Enhanced Ultrafiltration.

Water pollution is a major global issue, and antibiotic drugs released into aquatic environments by the pharmaceutical industry, such as ciprofloxacin, have negative consequences on both human health and the ecosystem. In this study, the performance of PVA as a polymer ligand for ciprofloxacin (CPFX) removal is evaluated through polymer-enhanced ultrafiltration using a novel composite PVC-ZnO membrane. The initial concentration of the ciprofloxacin solution, pH, ionic strength, ideal polymer concentration, duration, and maximum retention capacity were among the factors that were examined. In order to remove ciprofloxacin from water, PVA is utilized as a polymeric binding agent in a complex manufacturing process. In this instance, the PVC-ZnO membrane with 1.0 weight percent ZnO had a 96.77% ciprofloxacin clearance rate. PVA polymer has a high clearance rate of 99.98% in 1wt% of ZnO in this composite membrane when added to the ciprofloxacin solution. Fourier transform infrared spectroscopy (FTIR), energy dispersive X-ray spectroscopy (EDS), scanning electron microscopy (SEM), ultraviolet spectroscopy, and X-ray diffraction (XRD) were used to analyze the production and features of composite PVC-ZnO membranes. It is anticipated that this study's discussion will be crucial to the development of higher-quality membrane technologies that remove pharmaceutical active chemicals from wastewater in an environmentally responsible manner without endangering the ecosystem. This investigation showed that composite PVC-ZnO membranes were effective materials for efficient removal of ciprofloxacin (CPFX).

Impact of Chronic Inflammatory Diseaseson Clinical Pharmacokinetics of Antibody-Based Therapeutic Proteins.

Antibody-based therapeutic proteins (TPs) emerged as a promising therapeutic modality for chronic inflammatory diseases. During clinical development, the impact of disease on pharmacokinetics (PK) should be carefully considered to reliably extrapolate PK from healthy volunteers (HVs) in early-phase studies to patients in later-phase studies. This paper aimed to provide an overview of the impact of chronic inflammatory diseases on the PK of antibody-based TPs. A literature search was conducted on Drugs@FDA and PubMed, yielding 30 TPs with author-drawn conclusions about PK differences between HVs and patients with inflammatory diseases. Nine out of 30 TPs suggested PK differences in patients with inflammatory diseases compared with HVs, and patients mostly appeared to have higher drug clearance or lower drug exposure compared with HVs. However, the remaining 21 TPs appeared to have no apparent PK differences between patients and HVs. Based on ratios of reported drug clearance in patients vs. HVs (N: 31 ratios for 22 TPs), the difference in TP clearance due to inflammatory diseases did not exceed twofold (mean clearance ratio: 1.23; standard deviation: 0.25), with the most noticeable difference (>50% higher clearance) observed in patients with inflammatory bowel diseases and rheumatoid arthritis. Covariate assessment in published population PK models of TPs revealed that higher baseline C-reactive proteins and lower baseline albumin levels tend to be correlated with higher TP clearance. Future investigation is necessary to further elucidate the mechanism behind the inflammatory disease-mediated PK differences.