Months Of Dosing Research Articles

Efficacy of Lemborexant in Adults ≥ 65 Years of Age with Insomnia Disorder.

Pharmacologic treatments are available to treat insomnia, a common and burdensome sleep disorder, but may be contraindicated in older adults who are prone to side effects from sleep-promoting drugs. These analyses of sleep diary data from Study E2006-G000-303 (Study 303) investigated the benefits of lemborexant 5mg (LEM5) and 10mg (LEM10) in the subgroup age ≥ 65years with insomnia. Study 303, a 12-month, double-blind study of LEM5 and LEM10 in adults (age ≥ 18years) with insomnia disorder (sleep onset and/or maintenance difficulties) assessed subject-reported (subjective) sleep-onset latency (sSOL), sleep efficiency (sSE), wake after sleep onset (sWASO), and total sleep time (sTST). Morning sleepiness/alertness, insomnia severity (Insomnia Severity Index [ISI]), fatigue (Fatigue Severity Scale [FSS]), perceptions of sleep-related medication effects (Patient Global Impression-Insomnia [PGI-I] questionnaire), and safety were also evaluated. In this subgroup of older adults (≥ 65years; n = 262), there were significantly larger changes from baseline for sSOL, sSE, sTST, and sWASO with LEM5 and LEM10 versus placebo through month 6 (except sWASO month 1), indicating improvement; these improvements were sustained through month 12. Subject-reported increases in morning alertness were significantly greater with one or both LEM doses versus placebo through month 6 and sustained through month 12. There were significantly larger ISI total and daytime functioning score decreases (improvement) from baseline with LEM versus placebo at months 1, 3, and 6 (total score: both doses; daytime functioning: LEM5 month 1 and both doses months 3 and 6) and decreases from baseline FSS at months 1 and 3 (LEM5) and month 6 (both doses), sustained to month 12. Compared with placebo, more subjects reported that LEM (both doses) positively impacted ability to sleep, time to fall asleep, and TST through month 6, sustained to month 12, with no rebound after drug withdrawal. LEM was well tolerated to month 12; mild somnolence was the most common treatment-emergent adverse event. Improvements in subject-reported efficacy in LEM-treated adults age ≥ 65years with insomnia were observed as early as the first week of treatment and sustained through end of month 12. LEM was well tolerated. ClinicalTrials.gov identifier NCT02952820: E2006-G000-303; Study 303; SUNRISE-2 (First posted: October 2016); EudraCT 2015-001463-39 (First posted: November 2016).

Treatment of Hypothyroidism with Flavonoid Rich Extract of Fucus vesiculosus L Algae in Adult Rats

Background: Thyroid-related diseases, such as hypothyroidism, affect large populations worldwide and have severe health effects. Fucus vesiculosus L seaweed, which is full of iodine and other biologically active ingredients, has always been recognized as a remedy for hypothyroidism. The objective of this study was to explore the effects of a flavonoid extract of the algae, Fucus vesiculosus L, on the thyroid hormones in rat. Method: Thirty-two mature rats were separated into four categories and given different amounts of the seaweed extract over the span of a month. Blood samples were then taken and assessed for thyroid hormones, antioxidants, and liver enzymes. The effect of the flavonoid extract of the Fucus vesica algae on thyroid hormones in the serum of adult rats was studied, by estimation thyroid hormones, glutathione peroxidase, and peroxynitrite, in addition to liver enzymes in the blood serum of (32) adult rats. Results: The results showed that the TSH level a significant decrease in all groups compared to the control group after a month of dosing. T3 also showed a significant increase in all groups compared to the control, in addition to T4, which showed a significant increase in all groups. As for the level of glutathione peroxidase, it showed a significant decrease in all groups compared to the control group, while peroxynitrite showed a significant decrease in all groups compared to the control group. As for liver enzymes, they showed a significant decrease in all groups compared with the control group. Conclsuion: In summary, the Fucus vesiculosus L. seaweed flavonoid extracts showed a marked deviations in the concentrations of thyroid hormones and exposed potential hepato-protective and antioxidative activity in adult rats.

Quantifying non-adherence to anti-tuberculosis treatment due to early discontinuation: a systematic literature review of timings to loss to follow-up

BackgroundThe burden of non-adherence to anti-tuberculosis (TB) treatment is poorly understood. One type is early discontinuation, that is, stopping treatment early. Given the implications of early discontinuation for treatment outcomes,...

D-galactose Supplementation for the Treatment of Mild Malformation of Cortical Development with Oligodendroglial Hyperplasia in Epilepsy (MOGHE): A Pilot Trial of Precision Medicine After Epilepsy Surgery.

MOGHE is defined as mild malformation of cortical development with oligodendroglial hyperplasia in epilepsy. Approximately half of the patients with histopathologically confirmed MOGHE carry a brain somatic variant in the SLC35A2 gene encoding a UDP-galactose transporter. Previous research showed that D-galactose supplementation results in clinical improvement in patients with a congenital disorder of glycosylation due to germline variants in SLC35A2. We aimed to evaluate the effects of D-galactose supplementation in patients with histopathologically confirmed MOGHE, with uncontrolled seizures or cognitive impairment and epileptiform activity at the EEG after epilepsy surgery (NCT04833322). Patients were orally supplemented with D-galactose for 6months in doses up to 1.5g/kg/day and monitored for seizure frequency including 24-h video-EEG recording, cognition and behavioral scores, i.e., WISC, BRIEF-2, SNAP-IV, and SCQ, and quality of life measures, before and 6months after treatment. Global response was defined by > 50% improvement of seizure frequency and/or cognition and behavior (clinical global impression of "much improved" or better). Twelve patients (aged 5-28years) were included from three different centers. Neurosurgical tissue samples were available in all patients and revealed a brain somatic variant in SLC35A2 in six patients (non-present in the blood). After 6months of supplementation, D-galactose was well tolerated with just two patients presenting abdominal discomfort, solved after dose spacing or reduction. There was a 50% reduction or higher of seizure frequency in 3/6 patients, with an improvement at EEG in 2/5 patients. One patient became seizure-free. An improvement of cognitive/behavioral features encompassing impulsivity (mean SNAP-IV - 3.19 [- 0.84; - 5.6]), social communication (mean SCQ - 2.08 [- 0.63; - 4.90]), and executive function (BRIEF-2 inhibit - 5.2 [- 1.23; - 9.2]) was observed. Global responder rate was 9/12 (6/6 in SLC35A2-positive). Our results suggest that supplementation with D-galactose in patients with MOGHE is safe and well tolerated and, although the efficacy data warrant larger studies, it might build a rationale for precision medicine after epilepsy surgery.

A PAM of the α1A-Adrenergic receptor rescues biomarker, long-term potentiation, and cognitive deficits in Alzheimer’s disease mouse models without effects on blood pressure

α1-Adrenergic Receptors (ARs) regulate the sympathetic nervous system by the binding of norepinephrine (NE) and epinephrine (Epi) through different subtypes (α1A, α1B, α1D). α1A-AR activation is hypothesized to be memory forming and cognitive enhancing but drug development has been stagnant due to unwanted side effects on blood pressure. We recently reported the pharmacological characterization of the first positive allosteric modulator (PAM) for the α1A-AR with predictive pro-cognitive and memory properties. In this report, we now demonstrate the in vivo characteristics of Compound 3 (Cmpd-3) in two genetically-different Alzheimer’s Disease (AD) mouse models. Drug metabolism and pharmacokinetic studies indicate sufficient brain penetrance and rapid uptake into the brain with low to moderate clearance, and a favorable inhibition profile against the major cytochrome p450 enzymes. Oral administration of Cmpd-3 (3–9 mg/kg QD) can fully rescue long-term potentiation defects and AD biomarker profile (amyloid β-40, 42) within 3 months of dosing to levels that were non-significant from WT controls and which outperformed donepezil (1 mg/kg QD). There were also significant effects on paired pulse facilitation and cognitive behavior. Long-term and high-dose in vivo studies with Cmpd-3 revealed no effects on blood pressure. Our results suggest that Cmpd-3 can maintain lasting therapeutic levels and efficacy with disease modifying effects with a once per day dosing regimen in AD mouse models with no observed side effects.

Population Pharmacokinetic Analyses of Ravulizumab on-Body Delivery System for Subcutaneous Administration in Patients with Paroxysmal Nocturnal Hemoglobinuria

Background: The efficacy of ravulizumab (intravenous [IV] formulation administered every 8 weeks) for the treatment of paroxysmal nocturnal hemoglobinuria (PNH) and atypical hemolytic uremic syndrome (aHUS) has been demonstrated in randomized trials (NCT02946463, NCT03056040, NCT03406507, NCT02949128). Studies of IV ravulizumab have established the target therapeutic pharmacokinetic (PK) threshold of 175 μg/mL for complete and sustained terminal complement inhibition (free C5 < 0.5 µg/mL) in patients with PNH and aHUS; maintenance of serum ravulizumab trough concentration (Ctrough) above this threshold ensures clinical efficacy. In study 303 (NCT03748823), the on-body delivery system for subcutaneous (SUBQ) ravulizumab administration (490 mg self-administered weekly) showed PK non-inferiority to IV ravulizumab after 71 days of treatment in adult patients with PNH weighing ≥ 40 to < 100 kg who were clinically stable on prior IV eculizumab treatment (Yenerel et al. HemaSphere 2021;5:e566). In this study, a population PK (popPK) model was developed to characterize ravulizumab PK following IV or SUBQ ravulizumab treatment in patients with PNH, which was then used to generate model-based dosing simulations to support SUBQ and IV ravulizumab posology. Aims: To develop a popPK model and perform simulations to evaluate model-predicted 1) ravulizumab concentrations following IV and SUBQ administration over 1 year of treatment, 2) ravulizumab exposures following SUBQ administration in patients weighing ≥ 100 kg and 3) impact of switching administration route (from IV to SUBQ ravulizumab and vice versa). Methods: A popPK analysis of ravulizumab was performed using data from patients enrolled in three clinical trials (NCT05288829, NCT05396742, NCT03748823). Concentration-time profiles of ravulizumab following IV and SUBQ administration were simultaneously modeled with a 2-compartment model with linear clearance. Ravulizumab absorption following SUBQ administration was modeled using a first-order rate constant of absorption (Ka), bioavailability and lag time. Model-based analyses for repeated measures PK endpoints were conducted using nonlinear mixed effects modeling. The final popPK model accounted for covariates such as body weight, body mass index, disease status and hemoglobin, and was used to simulate steady state concentration profiles following dosing over 1 year, exposure in virtual patients weighing ≥ 100 kg and the impact of a switch in administration route after differing durations of dosing. Results: In total, 2689 blood samples (SUBQ: 1728; IV: 961) were obtained from 178 participants (129 with PNH and 49 without; 115 treated with SUBQ and 63 IV ravulizumab; 46.6% female; aged 18-79 years; body weight 43.5-98.4 kg). The mean (standard deviation) terminal elimination half-life of ravulizumab after SUBQ dosing was 52.4 (9.72) days. The bioavailable fraction of ravulizumab after SUBQ administration was 79.4%. The model-predicted geometric mean ratio (GMR) of SUBQ compared with IV Ctrough on Day 71 and 351 (steady state) were 1.213 and 1.392, respectively (Figure) and comparable to the Day 71 GMR of 1.257 in study 303, indicating the developed popPK model accurately characterized ravulizumab exposure for the ≥ 40 to < 100 kg body weight range in the trial. Simulations predicted that SUBQ ravulizumab Ctrough both remained above the target PK threshold and was non-inferior to ravulizumab IV for body weights ≥ 100 kg. Simulations describing a switch in ravulizumab formulation (IV to SUBQ or SUBQ to IV) show all concentrations remained within our target exposure range regardless of dosing order, after a single dose or after 6 months of dosing. Conclusions: A popPK model was developed and used to support an expanded indication and unrestricted dosing interchangeability between SUBQ and IV ravulizumab. Simulations using the popPK model predicted that switching SUBQ or IV ravulizumab administration route still maintained ravulizumab concentration within the target exposure range supporting treatment flexibility for patients receiving ravulizumab, and that the studied dose regimen maintained ravulizumab concentrations above the PK threshold for patients with body weight ≥ 100 kg. This study illustrates the utility of popPK modeling and simulation to supplement clinical trial results in expanding the target population and dosing options for patients with rare diseases. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Durability of Factor IX Activity and Bleeding Rate in People with Severe or Moderately Severe Hemophilia B after 5 Years of Follow-up in the Phase 1/2 Study of AMT-060, and after 3 Years of Follow-up in the Phase 2b and 2 Years of Follow-up in the Phase 3 Studies of Etranacogene Dezaparvovec (AMT-061)

Introduction: Gene therapy for people with hemophilia B appears to have a durable response, with data presented over 5 years (Samelson-Jones. Blood 2021; 138_suppl 1: 3975) and 8 years (Nathwani. Blood 2018; 132_suppl 1: 491) post-administration. Here we review the durability of AMT-060 and etranacogene dezaparvovec (formerly AMT-061) investigational gene therapies for hemophilia B, defined by sustained factor IX (FIX) activity levels and hemostatic protection. AMT-060 was developed as a precursor to etranacogene dezaparvovec. Both gene therapies are almost identical, comprising an adeno-associated virus serotype 5 (AAV5) vector containing a FIX transgene under the control of a liver-specific promoter; the transgene encodes wild-type FIX in AMT-060 and the highly active Padua variant of FIX in etranacogene dezaparvovec, differing by only one amino acid. Therefore, durability of response is expected to be similar for AMT-060 and etranacogene dezaparvovec. Aim: Outline the observed durability of AMT-060 and etranacogene dezaparvovec in people with severe or moderately severe hemophilia B. Methods: Three clinical trials are ongoing in adults with severe or moderately severe hemophilia B. Data have been captured over 5 years in the Phase 1/2 study assessing AMT-060 (N=10; Cohort 1: n=5, 5x1012 gc/kg ; Cohort 2: n=5, 2x1013 gc/kg; NCT02396342), 3 years in the Phase 2b study assessing etranacogene dezaparvovec (N=3, 2x1013 gc/kg; NCT03489291) and 2 years in the Phase 3 HOPE-B study assessing etranacogene dezaparvovec (N=54, 2x1013 gc/kg; NCT03569891). Study designs have been reported previously. All three studies assessed FIX activity levels and bleeding events. Results: In the Phase 1/2 AMT-060 study, FIX activity levels remained stable in both cohorts 5 years after dosing. Mean FIX activity was 4.4% for Cohort 1 at 52 weeks and 6.9% for Cohort 2 at 26 weeks; mean FIX activity after 5 years was 5.2% and 7.4%, for Cohorts 1 and 2, respectively. Greater FIX activity levels were achieved with etranacogene dezaparvovec. After 3 and 2 years of follow-up in the Phase 2b and 3 studies, respectively, mean FIX activity levels remained in the near-normal range (Table). In the Phase 2b study, mean FIX activity was 23.4% at Week 3 (n=3); this increased to 36.9% at Year 3 (n=2). A sustained mean FIX activity level was seen in the Phase 3 study: 38.95% at Month 6 (n=51) and 36.66% at Year 2 (n=50). This was observed in both the full analysis set (n=54) and in the modified intent-to-treat population (n=52) that excluded two non-responders (one with the highest AAV5 neutralizing antibody titer of 3212; one that received only a partial dose; Table). With regards to hemostatic protection, in Year 1 post-AMT-060 administration, the mean annualized bleeding rate (ABR) was 7.30 in Cohort 1, and this decreased to 6.40 at Year 5. In Cohort 2, mean ABR decreased from 1.58 in Year 1 to 0.20 in Year 5. The etranacogene dezaparvovec studies showed a similar low bleeding rate. In the Phase 2b study, no bleeding episodes occurred between 2.5-3 years of follow-up. Overall, the ABR over 3 years was 0.22. Two bleeding episodes (one very mild, spontaneous bleed; one traumatic) occurred in one participant within 18 months of dosing; the participant received FIX product replacement therapy (1700 IU) after each bleed. In the Phase 3 study, ABR (all bleeds) achieved at Months 7-18 post-administration was 1.51 (reduced from 4.18 at baseline); this was maintained through to 2 years in the full analysis set (n=54; Table). A similar trend was seen when assessing the modified intent-to-treat population (n=52), with a reduction in ABR from 4.00 at baseline to 0.95 after 2 years of follow-up. Reductions in ABR for spontaneous, joint, and traumatic bleeds were also maintained through 2 years of follow-up. No new safety signals have been identified after 5 years of follow-up for AMT-060 and up to 3 years of follow-up for etranacogene dezaparvovec. Conclusions: Gene therapy for hemophilia B appears to have a durable response. Across the Phase 1/2 study of AMT-060 and the Phase 2b and Phase 3 studies of etranacogene dezaparvovec, FIX activity levels are sustained throughout the observation period. Reductions in bleeding events also remained stable across the course of the three studies. Similar durable responses have been observed in published literature for hemophilia B gene therapies. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Assessment of moringa oleifera seeds extract effect on the adverse effects of levofloxacin drug induced liver injury

The current study aimed to know the protective role of the cold aqueous extract of Moringa seeds against the toxic damages caused by levofloxin in male albino rats by studying biochemical parameters. 24 male white rats were randomly divided into four groups, comprising (6 animals per group). The first group (G1) as a control group was dosed orally and daily with physiological solution for 30 days, the second group (G2)) was dosed with levofloxacin at a concentration of 10 mg / kg of body weight, the third group (G3) was dosed with aqueous extract of Moringa oleifera seeds at a concentration of 350 mg / kg The fourth group (G4) was dosed with aqueous extract of Moringa oleifera seeds 350 mg/kg 4 hours before the administration of levofloxacin at a concentration of 10 mg/kg body weight. Male rats in all study groups were administered orally via the dosing tube, with one dose per day for 30 days. . Blood samples were collected in the four groups after one month of dosing to measure the level of Malona dialdehyde and glutathione, as well as the levels of liver enzymes aspartate transaminase (AST), alanine transaminase (ALT) and alkalinephosphatase (ALP).

Pharmacological and clinical profile of Onasemnogene Aveparvovec, the first gene therapy for spinal muscular atrophy (SMA)

Onasemnogene abeparvovec (Zolgensma®; formerly AVXS-101) is a one-time gene therapy designed to address the genetic root cause of spinal muscular atrophy (SMA) by replacing the function of the missing or nonworking SMN1 gene via an adeno-associated AAV9 viral vector. On March 19, 2020, the Japanese Ministry of Health, Labor and Welfare approved onasemnogene abeparvovec for the treatment of SMA patients <2 years of age, including presymptomatic patients with a genetic diagnosis. Patients must be negative for elevated anti-AAV9 antibodies. Onasemnogene abeparvovec is administered through a single intravenous infusion, delivering a new working copy of the SMN gene into a patient's cells. Intravenous administration of onasemnogene abeparvovec to SMA model mice resulted in sustained expression of survival motor neuron (SMN) protein, weight gain, improvement of motor function, and prolongation of survival. Its clinical efficacy and safety have been demonstrated through the Phase I START and Phase III STR1VE-US, STR1VE-EU, and SPR1NT trials, and their long-term extension studies. SMA and presymptomatic patients treated with onasemnogene abeparvovec have achieved rates of survival not observed in the natural history of SMA. Treatment has led to rapid motor function improvement, often within one month of dosing, and developmental milestone achievement, including the ability to sit without support. The most commonly observed adverse effects after treatment were elevated liver enzymes, which often resolved with a course of prednisolone, and vomiting. This review discusses the rationale underlying gene replacement therapy for SMA, and describes the basic science, clinical trial experience, and use of onasemnogene abeparvovec.

Long-term use of Spirulina platensis, Ganoderma lucidum, silymarin and green tea can reduce the viral load and side effects of HCV patients

The present study aimed to evaluate the impact of the long-term treatment with a combination of Spirulina platensis (Sp), Ganoderma lucidum (Gd), silymarin (Sil), and green tea (Gt) in HCV patients. Forty-five adult males (M) and females (F) HCV patients’ elderly 40 to 65 were involved in this observational study. HCV patients were divided into 4 subjects; subject-1 included 15 HCV patients (10M and 5F) and was administered with INF/Rib for 12 months. Subject-2 contained 7M plus 3F dealt with Sp, Sil, and Gt. 10 HCV patients concerned in Subject-3 (7M and 3F) and handled with Sp, Sil, Gd, and Gt. Subject-4 covered 10 HCV patients (6M and 4F) who received Sp, Gd, and Gt. The combined treatments of Sp, Sil, Gd, and Gt were administered orally for 12 months in doses of 20, 4, 0.8, and 6 mg/kg, respectively. Viral load, hematological parameters, liver, kidney functions, and blood glucose were determined in all HCV patients. The results confirmed that each treatment condition did no longer dispose of viral particles in HCV patients contrasted to patients treated with INF/Rib. Furthermore, in subject three, the treatment with Sp, Sil, Gd, and Gt caused a potential decrease in the viral load, progression of hematological pictures, decreased levels of liver enzymes (ALT, AST), and urea. In conclusion, these treatments could be used as adjunct to conventional treatment regimens.

AL001 restores CSF PGRN levels and normalizes disease‐associated biomarkers in individuals with frontotemporal dementia due to heterozygous mutations in the progranulin gene

AbstractBackgroundAL001 is a recombinant humanized anti‐Sortilin monoclonal antibody in development for frontotemporal dementia (FTD). Mutations in a single copy of the progranulin gene (GRN) leads to a 50% or greater decrease in the level of circulating progranulin (PGRN) protein levels and are causative of FTD. The disease is associated with increased inflammation and impaired lysosomal function in the brain. The INFRONT Phase 1/1b clinical study AL001‐1 investigated the safety, tolerability, pharmacokinetics and pharmacodynamics of intravenously administered AL001 in healthy volunteers (HV) and participants with a GRN mutation causative of FTD (FTD‐GRN). In addition, the study investigated the effect of AL001 on disease‐associated biomarkers.MethodA total of 64 participants were enrolled in the AL001‐1 study. In the single dose ascending part of the study, 50 healthy volunteer participants were enrolled with 38 exposed to AL001 and 12 exposed to placebo. The study enrolled 6 asymptomatic FTD‐GRN and 8 symptomatic FTD‐GRN participants. The 8 symptomatic FTD‐GRN participants received 3 doses of 30 mg/kg of AL001 over 4 weeks.ResultTreatment of symptomatic FTD‐GRN participants with AL001 over 4 weeks restored PGRN levels in the cerebrospinal fluid (CSF) to normal levels for more than 8 weeks. To further investigate the effects of restoring PGRN levels, partial normalization of a FTD disease proteomic signature was observed in the CSF 4 weeks after the last dose in multidose treatment group. Compared to healthy volunteers, AL001 treatment rescued cathepsin B (CTSB), a marker of lysosomal function, by 58%. AL001 treatment also partially normalized markers of inflammation and gliosis, osteopontin (SPP1) and chitotriosidase (CHIT1) by 52% and 22%, respectively. In addition to the reversal of these disease phenotypes, a 14% reduction (n.s.) in plasma neurofilament light chain (NfL), a marker of neuron injury and stress, was observed after 8 weeks after the last dose.ConclusionAL001 treatment resulted in a sustained increase in plasma and CSF PGRN in FTD‐GRN mutation carriers. New data showed reduction of NfL and normalization of proteins associated with the disease after one month of dosing. These results have supported progressing AL001 to Phase 2 and pivotal Phase 3 studies.

Influence of additive components with separate introduction of antagonistic components on gastrointestinal microorganization and microelements content in mouse blood serum

Taking into account the data on higher biological availability of chelate compounds of microelements in comparison with their inorganic salts, a liquid feed additive for cattle with the content of fat-soluble vitamins A, D, E and diacetophenonyl selenide (DAFS 25) in the form of stable microemulsion and chelates of manganese, iron, cobalt, and zinc was developed. The biochemical and physiologically antagonistic components of the additive are divided into two separate liquid mixtures for separate, alternating ingestion on white laboratory mice, histological methods evaluated the effect of different doses of the additive on the micro organization of the components of the stomach wall, small and large intestine in different terms of the experiment. The additive was introduced into animals individually daily per o s during one month in doses of 0.06, 0.12, 0.3, 0.6 and 1.2 g/kg. The control animals received an equivalent amount of physiological solution. The availability of trace elements of the developed product is estimated by analysis of blood serum on the zinc, iron, manganese, cobalt content by atomic-adsorption spectrometry. Examination bodies and blood for analysis were selected at the time of slaughter in accordance with generally accepted requirements. The dosage of the additive, which has the most effective influence on the growth of the concentration of injected micronutrients in the serum of mice in comparison with the control, was determined. It was found that the components of the additive have the most pronounced effect on zinc and iron content in the dose of 0.3 g/kg.

Rapid and Sensitive Quantification of Osimertinib in Human Plasma Using a Fully Validated MALDI-IM-MS/MS Assay.

The third-generation tyrosine kinase inhibitor (TKI), osimertinib, has revolutionized the treatment of patients with non-small cell lung carcinoma with epidermal growth factor receptor (EGFR)-activating mutation, and resistant to first- and second-generation TKIs. Osimertinib is now also proposed as a first-line therapy, thus extending the scope of applications in lung oncology. Personalized medicine approaches are still necessary to monitor if patients are exposed to adequate concentrations of osimertinib during their treatment. It would also help to understand the appearance of new resistances in patients after several months of dosing with osimertinib. Liquid chromatography–tandem mass spectrometry (LC–MS/MS) is currently the gold standard for the quantification of drugs in plasma enabling pharmacokinetic analyses and patient monitoring. In the present study, we propose an alternative to LC–MS/MS methods for the rapid and sensitive quantification of osimertinib in plasma using matrix-assisted laser desorption/ionization (MALDI) –MS. The presented assay requires only 3 min per sample for their preparation, analysis, and data extraction, and less than 3 h for quantification. A lower limit of quantification (LLOQ) of 5 ng/mL in plasma was retrieved. The method was fully validated, following the guidelines of the US Food and Drug Administration (FDA) and the European Medicines Agency (EMA) for bioanalytical method validation. The present developments prove the importance to consider alternative MS assays for time-efficient quantification of small molecule inhibitors in plasma in the context of personalized medicine for targeted therapies.

1532. Population Pharmacokinetic (PPK) Modeling and Simulation of Long-Acting (LA) Cabotegravir (CAB) to Inform Strategies Following Dosing Interruptions in HIV-1-Infected Subjects

BackgroundCAB is an integrase strand transfer inhibitor under investigation as an injectable LA formulation for the treatment and prevention of HIV, and as a tablet formulation as an oral lead-in (OLI) and bridging treatment for dose interruptions. The monthly injection regimen of CAB LA and rilpivirine (RPV) LA was noninferior to standard oral therapy in maintaining HIV-1 suppression in Phase 3 trials. PPK modeling and simulation was used to inform strategies for managing dosing interruptions.MethodsA 2-compartment model with first-order oral and LA absorption and elimination adequately described the data from 1,647 healthy (28%) and HIV-infected (72%) adult subjects in 16 studies. Gender was a significant covariate on LA absorption; therefore, simulations of 5,000 virtual subjects were performed using a 4:1 male:female ratio to ensure 1,000 representative females and covariate sampling with replacement from the analysis dataset. One- to 12-week delays in dosing of the second, third, and fourth injection were simulated, and predicted troughs were compared with the 5th percentile (0.65 μg/mL) of trough concentrations following the first injection in Phase 3. Simulations of 1–2 months of oral bridging with CAB 30 mg once daily from time of a missed injection until CAB LA dosing resumed were performed, with the median Cmax (13.1 μg/mL) observed following oral CAB 60mg once daily in Phase 2b as an upper reference.ResultsProportions of subjects predicted to achieve target plasma CAB trough concentrations are shown by length of delay and injection visit in Table 1. Oral bridging with CAB 30mg once daily starting at the time of a planned missed injection is predicted to provide exposures within ranges observed in clinical studies (Figure 1).ConclusionDosing delays of up to one week appear to have minimal impact, but the effect is more likely to become problematic with longer delays, particularly in the first few months of dosing. Oral bridging provides therapeutic and safe exposures for planned interruptions in LA dosing. Regardless of use of oral bridging, simulations support resuming CAB LA dosing for interruptions <1 month (<2 months between injections) and reinitiating CAB LA with a loading dose and subsequent monthly injections for interruptions ≥ 1 month (≥ 2 months between injections). Disclosures All authors: No reported disclosures.

Lessons learned from the metastatic castration-resistant prostate cancer phase I trial of EPI-506, a first-generation androgen receptor N-terminal domain inhibitor.

257 Background: Aniten compounds bind to the N-terminal domain (NTD) of the androgen receptor (AR) and inhibit AR dependent transcription. EPI-506, the pro-drug of EPI-002, was the first AR NTD inhibitor tested in a Phase 1 study in men with metastatic castration-resistant prostate cancer (mCRPC). The drug was well-tolerated but required high doses. At doses &gt;1280 mg, EPI-506 treatment resulted in PSA declines; however, these were minor and of short duration, reflecting EPI-506’s low potency and short half-life. To understand EPI-506’s metabolic vulnerabilities, patient plasma samples were analyzed to identify metabolites. Methods: PSA serum levels were assessed after a month of dosing. Patient plasma samples were analyzed and pharmacokinetic (PK) parameters calculated. Three plasma samples from patients (one 80 and two 3,600 mg doses), were pooled across timepoints and metabolites were analyzed. EPI-506 metabolism was assessed in in vitro ADME assays and metabolite activity was measured. Results: EPI-002 patient plasma profiles exhibited dose-proportional Cmax and AUC following once or twice-daily EPI-506 administration. PSA declines (range of 8-29%) were observed, especially at higher doses (≥ 1,280 mg). A total of 19 metabolites were identified. Metabolite M19, a glycerol-moiety oxidant, was the major drug-related component. Other metabolic pathways included O-glucuronidation, sulfation, carboxylic acid formation, and oxidative chlorine loss. The major metabolites were tested in an AR driven reporter assay and were shown to be inactive. Interestingly, in vitro ADME assays predicted glucuronidation and sulfation but not cytochrome dependent metabolism. Conclusions: EPI-506 was tested in a phase 1 trial and showed minor PSA declines. The drug was well-tolerated but was highly metabolized. Patient plasma samples identified 19 metabolites. Newer molecules have been synthesized to address EPI-002’s metabolic liabilities and demonstrate &gt; 20-fold improved potency and higher stability. These next generation Anitens are currently being characterized for IND filing. Clinical trial information: NCT02606123.

Phase 1 Trial of Cobomarsen, an Inhibitor of Mir-155, in Cutaneous T Cell Lymphoma

Background:Cobomarsen (MRG-106) is an inhibitor of miR-155, a microRNA with a strong link to cutaneous T cell lymphoma (CTCL) pathogenesis. The goals of this first in human study are to evaluate the safety, tolerability, pharmacokinetics, and efficacy of cobomarsen in mycosis fungoides (MF) patients.Methods:This Phase 1 trial evaluated cobomarsen given via intralesional injection (75 mg/dose), subcutaneous (SC), IV rapid bolus injection, or 2-hour IV infusion (300, 600 or 900 mg/dose). Patients must have MF stage I-III with plaques and/or tumors and could remain on concurrent stable CTCL therapy. Patients received 6 doses, either subcutaneous or intravenous, in the first 26 days of the study followed by weekly or bi-monthly doses. Safety was monitored by physical exams, clinical lab tests, and reported adverse events (AEs). Efficacy was assessed by CAILS and by the modified Severity Weighted Assessment Tool (mSWAT). The effect of cobomarsen on quality of life was assessed by Skindex-29.Results:38 subjects receiving IV or SC treatment (25 male/13 female, median age 59 years) have been on study for up to 22 months. As of the data cut off, no serious AEs have been attributed to cobomarsen. The most common AEs reported in greater than 15% of subjects were: fatigue, neutropenia, injection site pain, nausea, pruritus, and headache. Two AEs were deemed dose limiting toxicities (DLTs): Grade 3 worsening pruritus and Grade 3 tumor flare. The maximum tolerated dose (MTD) has not yet been reached.In the subcutaneous and IV cohorts, 29 out of 32 (91%) evaluable subjects had improvement in mSWAT score. Skin improvements were observed as early as the first assessment (Day 17). The best improvements were observed after more than 1 month of treatment. Eleven out of twenty-one (52%) patients receiving more than one month of dosing (6 doses) achieved greater than 50% reduction in mSWAT score. The mean duration of response (n=11) was 213 days, as of data cut off. Eight patients achieved a partial response meeting the criteria for ORR4, an objective response rate lasting at least four months in duration.The overall skin response in patients who received cobomarsen as monotherapy or cobomarsen with concurrent stable therapy were not significantly different. Improvement in quality of life (QOL), as measured by the Skindex-29 total score, correlated with reductions in mSWAT score during the treatment phase.Conclusions:These results demonstrate that cobomarsen is well-tolerated, has clinical activity, and has the potential to impact MF quality of life. These encouraging data support the continued investigation of cobomarsen in the CTCL population. The study is expanded to enroll patients with other hematologic malignancies in which miR-155 is elevated and relevant, including chronic lymphocytic leukemia (CLL), diffuse large B cell lymphoma (DLBCL), and adult T cell lymphoma/leukemia (ATLL). Final safety and efficacy data on CTCL mycosis fungoides will be presented. DisclosuresFoss:Seattle genetics: Consultancy; Miragen: Consultancy, Speakers Bureau; Spectrum: Consultancy; Mallinkrodt: Consultancy.

Analysis of Initial Nonresponders to Galcanezumab in Patients With Episodic or Chronic Migraine: Results From the EVOLVE-1, EVOLVE-2, and REGAIN Randomized, Double-Blind, Placebo-Controlled Studies.

ObjectiveTo examine the likelihood of response with continued galcanezumab treatment in patients with episodic or chronic migraine without initial clinical improvement.BackgroundA percentage of patients with migraine may require additional time on pharmacotherapy but discontinue treatment prematurely. Additionally, recognizing when continued treatment is unlikely to provide improvement limits unnecessary exposure.MethodsPost hoc analysis of response after continued galcanezumab treatment was conducted in a subset of patients with episodic (N = 879) and chronic (N = 555) migraine who did not achieve “good” early improvement (episodic, ≥50% reduction in baseline migraine headache days [MHD] and chronic, ≥30% reduction) after 1 month of dosing (NR‐1; episodic, n = 450 and chronic, n = 306). This subset was categorized by level of reduction in MHD during 1 month of treatment: “modest” (>30% to <50% fewer MHD for episodic and >10% to <30% fewer MHD for chronic), “limited” (episodic only; >10% to ≤30% fewer MHD), or “minimal/no” early improvement (≤10% fewer MHD to ≤10% more MHD), or “worsening” (>10% more MHD). The percentages of patients having “better” (≥75% fewer MHD for episodic and ≥50% for chronic), “good,” or “little‐to‐no” (≤10% fewer MHD) response during the remaining treatment period were calculated for each category. Similarly, the subset of NR‐1 patients who did not achieve “good” early improvement after 2 months of treatment (NR‐2; episodic, n = 290 and chronic, n = 240) were categorized by level of their average monthly reduction across 1 and 2 months using similar categories.ResultsOf NR‐1 patients with episodic migraine having “modest” early improvement, 62% (96/155) achieved “good” and 20% (31/155) achieved “better” responses with continued treatment. A percentage of patients with “limited” (43%; 46/108) or “minimal/no” (34%; 29/85) early improvement, or “worsening” (20%; 20/102) achieved a “good” response after continued treatment. A percentage of NR‐1 patients with chronic migraine having “modest” early improvement achieved “good” (38%; 44/116) and “better” (13%; 15/116) responses with continued treatment. A “good” response was achieved for a percentage of patients with “minimal/no” early improvement (17%; 23/133). Similar patterns were observed for the NR‐2 subset, though percentages were lower.ConclusionsGalcanezumab‐treated patients with episodic or chronic migraine without response following 1 or 2 months of treatment appear to have a reasonable likelihood of continued improvement in months following initial treatment and this opportunity is more likely in patients showing greater early improvements. While a small percentage of patients with episodic or chronic migraine who experienced worsening in the number of MHD following initial treatment responded with continued treatment, most do not show substantial reduction in MHD. Overall benefit of therapy should be determined collaboratively between the patient and physician.

THE ASSESSMENT OF THE ANTIOXIDANT STATUS OF THE ORGANISM OF RATS UNDER THE CHRONIC INTAKE OF SURFACE-ACTIVE SUBSTANCES BASED ON THE RAPE OIL

Introduction. The study of the character of reactions of lipid peroxidation, as well as the state of the antioxidant system regulating their course under the conditions of the action of chemical products on the body, is an important stage of investigations. There is not enough data on the development of oxidative stress in cells exposed to pesticides, and this concerns, first of all, studies of the activity of antioxidant enzymes. The aim of the study is to execute the experimental assessment of the influence of a surface active substances (SAS) on the basis of the rape oil on antioxidant status of white male rats during its multiple peroral intakes for 6 months in doses of 0.5, 5.0 and 50.0 mg/kg body weight (on a.s.). Material and methods. Studies on the determination of enzymatic indices of the body's antioxidant defense system (superoxide dismutase, glutathione peroxidase and glutathione reductase) were performed on a ChemWell biochemical analyzer (Awareness Technology, USA) with the use of diagnostic reagent kits by Randox Laboratories Ltd (England). The catalase activity was determined by the colorimetric method. Results. Under the prolonged action of surfactants on warm-blooded organisms, changes in the activity of antioxidant enzymes were established to occur in doses of 5,0 and 50,0 mg/kg b.m. Discussion. The test substance used in a dose of 0.5 mg/kg bm does not have an inhibitory effect on the antioxidant status of the organism. The use of a surfactant based on rapeseed oil at doses of 5.0 and 50.0 mg/kg mt has a negative effect on the antioxidant status of an organism of male rats. Conclusion. Surfactants based on rapeseed oil in doses of 5.0 and 500 mg/kg b.m. have a negative effect on the antioxidant status of an organism of male rats, which is probably due to the high content of erucic acid and glucosinolates in rapeseed oil.

Abstract LB-B33: Nonclinical safety evaluation of two distinct second generation variants of anti-CTLA4 monoclonal antibody, ipilimumab, in monkeys

Abstract Ipilimumab is a fully human immunoglobulin G1 (IgG1) monoclonal antibody against cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), an inhibitory receptor expressed on activated effector T cells and regulatory T cells (Treg), that inhibits the binding of CTLA4 to B7 ligands. BMS-986218 and BMS-986249 are second generation molecules that share the same amino acid sequence and ligand blocking properties as ipilimumab, but are mechanistically distinct. BMS-986218 is non-fucosylated (NF) and has an increased affinity for the activating Fcγ receptor (FcγR, CD16) affording the possibility of increased anti-tumor activity via depletion of Treg in the tumor. In contrast, BMS-986249 is a Probody™ of ipilimumab that has a masking peptide covering the active antigen-binding site of the antibody which is clipped by specific proteases within tumors, exposing the fully active antibody, and potentially offering reduced systemic toxicity liabilities with comparable efficacy to ipilimumab. In 1-month toxicity studies in monkeys (n = 5/sex/group) at weekly doses of 3, 15, or 75 mg/kg IV of BMS-986218 or 10 or 50 mg/kg IV of BMS-986249 or ipilimumab, profound enhancement of peripheral T-cell activation occurred in a dose-dependent manner for all 3 compounds following neoantigen immunization (keyhole limpet hemocyanin [KLH], HIV necessary and enforcing factor [Nef], and HIV group specific antigen [Gag] peptides), consistent with target pharmacology. Consistent with the intended mechanistic differences, peripheral T cell activation was generally increased at corresponding doses of BMS-986218 compared to ipilimumab, and was delayed and reduced in monkeys given BMS-986249. BMS-986218, ipilimumab, and BMS-986249 were clinically tolerated by monkeys at doses up to 3, 10, and 50 mg/kg, respectively, with generally mild, loose feces in some monkeys and/or minimal body weight decrease. At higher doses, early euthanasia occurred for 1 and 6 monkeys at 15 and 75 mg/kg BMS-986218, respectively, from Days 22-53 and 1 monkey at 50 mg/kg ipilimumab on Day 55 due to profound clinical toxicity. The predominant microscopic finding was generally dose-related lymphohistiocytic inflammation within a variety of tissues at all doses for all compounds, with BMS-986218 resulting in the greatest incidence, severity, and distribution of tissues and BMS-986249 having the least effects. The GI tract (stomach, cecum, and colon) and the kidney were the most severely and consistently affected, whereas additional organs were affected at higher doses. Most changes were partially or fully reversible during an 8-week recovery period with the exceptions of one monkey at 75 mg/kg BMS-986218 and one monkey at 50 mg/kg ipilimumab that were euthanatized on Days 53 or 55 due to unresolved GI toxicity, persistent lymphohistocytic inflammation, and/or unchanged or progressive increases in AST and ALT. Based on the tolerability and generally mild severity of lymphohistiocytic tissue inflammation, the highest non-severely toxic doses (HNSTD) for BMS-986218, ipilimumab, and BMS-986249 in monkeys following 1 month of dosing were 3 mg/kg (mean AUC 0-168h = 11,300 μg•h/mL), 10 mg/kg (AUC[0-168h] =44,600 µg·h/mL), and 50 mg/kg (AUC[0-168h] =205,000 µg·h/mL), respectively. Overall, these results support the potential of these 2nd generation anti-CTLA4 antibodies to offer an improved risk /benefit profile with increased activity of the NF variant and improved safety of the Probody™ relative to ipilimumab. Citation Format: Karen D. Price, Frank Simutis, Anthony Fletcher, Lila Ramaiah, Rima Srour, John Kozlosky, Jean Sathish, John Engelhardt, Annette Capozzi, James Crona, Courtni Newsome, Jennifer Wheeler, Daniel Szatkowski, Austin Thekkumthala, Bojing Wang, Wendy Freebern, Helen Haggerty, Todd Bunch, Michael Graziano. Nonclinical safety evaluation of two distinct second generation variants of anti-CTLA4 monoclonal antibody, ipilimumab, in monkeys [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2017 Oct 26-30; Philadelphia, PA. Philadelphia (PA): AACR; Mol Cancer Ther 2018;17(1 Suppl):Abstract nr LB-B33.

Remediation of incomplete nitrification and capacity increase of biofilters at different drinking water treatment plants through copper dosing.

Drinking water treatment plants based on groundwater may suffer from incomplete ammonium removal, which deteriorates drinking water quality and constrains water utilities in the operation of their plants. Ammonium is normally removed through nitrification in biological granular media filters, and recent studies have demonstrated that dosing of copper can stimulate the removal of ammonium. Here, we investigated if copper dosing could generically improve ammonium removal of biofilters, at treatment plants with different characteristics. Copper was dosed at ≤1.5 μg Cu/L to biofilters at 10 groundwater treatment plants, all of which had displayed several years of incomplete nitrification. Plants exceeded the Danish national water quality standard of 0.05 mg NH4+/L by a factor of 2-12. Within only 2-3 weeks of dosing, ammonium removal rates increased significantly (up to 150%). Nitrification was fully established, with ammonium effluent concentrations of <0.01 mg NH4+-N/L at most plants, regardless of the differences in raw water chemistry, ammonium loading rates, filter design and operation, or treatment plant configuration. However, for filters without primary filtration, it took longer time to reach complete ammonium removal than for filters receiving prefiltered water, likely due to sorption of copper to iron oxides, at plants without prefiltration. With complete ammonium removal, we subjected two plants to short-term loading rate upshifts, to examine the filters' ability to cope with loading rate variations. After 2 months of dosing and an average loading rate of 1.0 g NH4+-N/m3 filter material/h, the loading rate was upshifted by 50%. Yet, a filter managed to completely remove all the influent ammonium, showing that with copper dosing the filter had extra capacity to remove ammonium even beyond its normal loading rates. Depth sampling revealed that the ammonium removal rate of the filter's upper 10 cm increased more than 7-fold from 0.67 to 4.90 g NH4+-N/m3/h, and that nitrite produced from increased ammonium oxidation was completely oxidized further to nitrate. Hence, no problems with nitrite accumulation or breakthrough occurred. Overall, copper dosing generically enhanced nitrification efficiency and allowed a range of quite different plants to meet water quality standards, even at increased loading rates. The capacity increase is highly relevant in practice, as it makes filters more robust towards sudden ammonium loading rate variations.