Single-dose Administration Study Research Articles

Risk of unintentional antidoping rule violations by consumption of hemp products.

Consumption of hemp products is continuously growing, with an expanding scope of applications. Suppliers operate through different distribution channels, but the Internet is a major retail platform. Hemp products are prepared from cannabis plants and, therefore, might contain a variety of different natural cannabinoids. According to the regulations of the World Anti-Doping Agency, all natural and synthetic cannabinoids are prohibited in-competition, with the explicit exemption of cannabidiol. Therefore, an investigation of 23 hemp products for the presence of cannabinoids was performed to determine the likelihood of unintentional violations of anti-doping regulations. An assay for the detection of 16 cannabinoids in nutritional supplements was developed and validated. The sample preparation consisted of QuEChERS extraction, trimethylsilylation, and analysis by gas chromatography/tandem mass spectrometry. All 23 commercially available hemp products were analyzed, and assay characteristics such as selectivity, limit of detection, limit of identification, limit of quantification, linearity, imprecision, recovery, and accuracy were determined. Twenty of 23 hemp products included a variety of cannabinoids at, occasionally, substantial concentrations, with four products covering the entire spectrum of tested cannabinoids. An ethics committee-approved single-dose administration study was conducted with the commercially available hemp products, investigating the presence of 16 cannabinoids in urine collected pre- and post-consumption. Variable patterns of cannabinoids or their metabolites in urine were observed. In 30% of the urine samples collected 8h after ingestion, the presence of a prohibited cannabinoid would have resulted in an unintentional violation of anti-doping regulations.

A Novel Anti-IgM/HLA-DR Bispecific Antibody for Treatment of Refractory B Cell Malignancies

Introduction: IgM composes a part of the B cell receptor, and its expression is restricted to B cell lineage cells. There have been several reports that refractory B cell malignant cells frequently express IgM. For example, activated B cell-like (ABC) diffuse large cell lymphoma (DLBCL) is a subtype with a poor prognosis following R-CHOP treatment (Lenz et al., N Engl J Med, 2008), and the tumor cells generally express IgM-type immunoglobulin (Ruminy et al., Leukemia, 2011). In addition, IgM-expressing chronic lymphocytic leukemia (CLL) was also found to have a poor prognosis (D'Avola et al., Blood, 2016). Therefore, IgM is a promising therapeutic target for refractory B cell malignancies. However, anti-IgM antibodies may not function fully in the body due to the presence of soluble IgM. To improve the selectivity to the IgM expressing cells, we produced a bispecific antibody bound to IgM and HLA-DR, which are both expressed by B cells, and it was able to bind IgM+HLA-DR+ cells in the presence of soluble IgM. Thus, the aim of the present study was to examine the in vitro anti-tumor activity of the anti-IgM/HLA-DR bispecific antibody against B cell lymphoma, and to clarify its efficacy and tolerability in non-human primates (NHP).Methods: BTA106 is the bispecific antibody that can bind to IgM and HLA-DR, and BTA111 is glycoengineered BTA106. To investigate their direct inhibitory effects and Fc-mediated effects against B cell lymphoma cell lines, a growth inhibition assay, antibody-dependent cell cytotoxicity (ADCC) assay and complement dependent cytotoxicity (CDC) assay were performed. In the growth inhibition assay and CDC assay, cell proliferation was measured by WST-8. In the ADCC assay, human peripheral blood mononuclear cells were used as effector cells. Dead cells were stained with propidium iodide and measured by flow cytometry. JeKo-1 (Mantle cell lymphoma, MCL), Ramos (Burkitt's lymphoma, BL), RL (DLBCL), B104 (B cell lymphoma) and two rituximab-resistant cell lines, RL-4RH (Czuczman et al., Clin Cancer Res., 2008) and RRBL1 (generously provided by Dr. Tomita, Tomita et al., Int J Hematol., 2007), were used. To clarify the efficacy and tolerability of BTA106 in NHP, a dose escalation study and single-dose administration study were performed using cynomolgus monkeys.Results: BTA106 and BTA111 demonstrated direct growth inhibition activity, and CDC and ADCC activity, although their intensities differed among cell lines. In the case of rituximab-resistant cell lines, RRBL1 and RL-4RH, BTA106 and BTA111 exhibited weak or no growth inhibitory effects, but their CDC and ADCC activity was superior to that of rituximab. Development of resistance to rituximab is also a poor prognostic factor and remains problematic. Thus, BTA106 and BTA111 are expected as therapeutic antibodies to overcome rituximab resistance.A dose escalation study and single dose study were performed in cynomolgus monkeys. BTA106 greatly reduced peripheral B cells. Moreover, after the administration of BTA106, lymph follicles disappeared in axillary lymph nodes. In these studies, decreased locomotor activity and vomiting were observed; however, these symptoms resolved 2 hrs after injection. Other BTA106-related clinical toxicities or changes in body weight were not observed.Conclusions: In the in vitro studies, BTA106 and BTA111 demonstrated significant anti-tumor effects on rituximab-sensitive and -resistant B cell lymphoma cell lines. The efficacy and toxicity studies in cynomolgus monkeys revealed that BTA106 depletes peripheral and lymph node B cells, and it was well tolerated. BTA111 was considered to be significantly more effective than BTA106 due to glycoengineering, which induced high ADCC activity. These data suggest that BTA111 is an effective and potent therapeutic antibody for B cell malignancies, especially rituximab-resistant B cell lymphoma. Currently, a repeated administration study of BTA111 with cynomolgus monkeys to evaluate its efficacy and toxicity is ongoing. DisclosuresOhashi:Zenyaku Kogyo Co., Ltd.: Employment. Miyashita:Zenyaku Kogyo Co., Ltd.: Employment. Nagata:Zenyaku Kogyo Co., Ltd.: Employment. Tatebe:Zenyaku Kogyo Co., Ltd.: Employment. Otsuka:Zenyaku Kogyo Co., Ltd.: Employment. Suzuki:Zenyaku Kogyo Co., Ltd.: Employment. Sasaki:Zenyaku Kogyo Co., Ltd.: Employment. Enami:Zenyaku Kogyo Co., Ltd.: Employment. Tsukada:Zenyaku Kogyo Co., Ltd.: Employment. Fukushima:Zenyaku Kogyo Co., Ltd.: Employment.

Abstract 2607: Endoxifen pharmacokinetics and bioavailability of in female rats and dogs

Abstract Endoxifen (END), the active metabolite of tamoxifen (TAM), is currently being developed as a drug for the treatment of estrogen receptor positive breast cancer based on recent in vitro and clinical data demonstrating that TAM drug effect is substantially related to circulating plasma concentrations of END, a secondary metabolite produced by human CYP2D6 metabolism. Women with deficient CYP2D6 metabolism have lower END concentrations and a higher risk of breast cancer recurrence. We characterized the pharmacokinetics and oral bioavailability of END in female rats and dogs. We further compared END exposure following equivalent doses of END and TAM. Rats were administered 2 mg/kg i.v. END, 20 − 200 mg/kg oral END or 80 mg/kg oral TAM. Fasted dogs were administered 0.5 mg/kg i.v. END, 15 − 100 mg/kg oral END or 30 mg/kg oral TAM. Plasma END concentrations were also measured in rats or dogs after oral dosing once per day for 4 consecutive days. Plasma samples were analyzed using a validated reverse-phase HPLC method with fluorescence detection. In the single-dose administration study, terminal elimination half-life and plasma clearance values in rats given 2 mg/kg i.v. END were 6.3 h and 2.4 L/h/kg, respectively. Plasma concentrations above 0.1 µM and 1 µM END were achieved with 20 mg/kg and 200 mg/kg doses, respectively, and those concentrations were maintained for longer than 24 h. Oral absorption of END was linear over the range of 20 − 140 mg/kg with a bioavailability greater than 67%. END exposure in rats following 80 mg/kg oral END was 100-fold greater than END exposure following 80-mg/kg oral TAM. Following 0.5 mg/kg i.v. END to dogs, terminal elimination half-life and plasma clearance values were 9.2 h and 0.4 L/h/kg, respectively. Plasma concentrations above 1 µM and 10 µM were achieved with the 15 mg/kg dose and 100 mg/kg, respectively, and those concentrations were maintained for longer than 24 h. Absorption was linear over the range of 15 − 100 mg/kg with a bioavailability greater than 50%. END exposure in dogs following 30 mg/kg oral END was 10-fold greater than END exposure following 30-mg/kg oral TAM. In the repeated-dose administration study, the day 4 END peak plasma concentration in rats increased from 1 to 9 µM as the daily END dose was increased from 20 to 200 mg/kg. The day 4 END peak plasma concentration in dogs exceeded 20 µM and plasma concentrations remained above 2 µM for 24 hours following 4 daily doses of 30 mg/kg END. Based on these data, we conclude that END has high oral bioavailability; high concentrations may be maintained after repeated dosing in dogs and rats; and substantially higher END concentrations are achievable following an oral dose of END in comparison to a similar dose of TAM. Supported by NCI contract N01-CM-52206. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2607.

Dose escalation trial of the efficacy, safety, and pharmacokinetics of a novel fibrinolytic agent, BB-10153, in patients with ST elevation MI: Results of the TIMI 31 trial

Currently available fibrinolytic agents are limited by their ability to restore normal blood flow in only half of patients, the risk of reocclusion, and the risk of intracranial hemorrhage. The genetically engineered agent BB-10153 is activated by thrombin, not plasminogen activator enzymes, which limits its activity to the site of thrombus which may in turn reduce the risk of systemic bleeding. BB-10153 also has a relatively long half-life of 3-4 hours, which may also limit the potential for early reocclusion [1, 2]. The study was a phase II, open-label, multi-center, dose escalation, single-dose administration study to determine the efficacy, safety, tolerability, pharmacokinetics and pharmacodynamics of BB-10153 in ST segment elevation MI (STEMI). STEMI patients (n = 50) received a single dose of BB-10153 at one of six dose levels (1.0, 2.0, 3.0, 5.0, 7.5 and 10 mg/kg). The primary endpoint was TIMI flow grade (TFG) 3 at 60 minutes following the intravenous bolus of BB-10153. Mean area under the curve for drug concentration ranged from 48.0 microg.h/mL in the 1 mg/kg dose group to 788.6 microg.h/mL in the 10 mg/kg dose group. Likewise, mean Cmax generally increased with dose over the entire dose range, from 4.9 microg/mL in the 1 mg/kg dose group to 139.6 microg/mL in the 10 mg/kg dose group. The mean apparent terminal half-life (t1/2) was 4.4 hours (range 2.2 to 7.6 hours). Few patients in the 1-3 mg/kg dosage groups achieved TFG 3 on the one-hour post-dose angiogram (4/20, 20%), and no patients achieved complete ST segment resolution. The 5, 7.5 and 10 mg/kg doses were associated with similar rates of TIMI grade 3 flow of approximately three per seven patients. Pooling TFG 3 data from the 5, 7.5 and 10 mg/kg groups yielded a TIMI grade 3 flow rate of 34% (n = 10/29; range 29-43%). No patients experienced 30-day death, recurrent acute MI, cardiogenic shock, stroke or anaphylaxis during the study. One patient experienced recurrent angina and developed recurrent myocardial ischemia requiring urgent revascularization. Three patients sustained TIMI major bleeding events (one in 1 mg/kg group, two in 7.5 mg/kg group), six patients sustained TIMI minor bleeds (one in the 2, 3, 7.5 and 10 mg/kg groups, two in the 5 mg/kg group), twp patients sustained TIMI minimal bleeds (one in each of the 2 and 10 mg/kg groups) and no patients sustained intracranial hemorrhage (ICH). In a dose escalation study of a single intravenous bolus, the novel fibrinolytic agent, BB-10153 was associated with a rise in the mean area under the curve and Cmax for drug concentration over the dose range 1 to 10 mg/kg. Higher doses were associated with a range of TIMI grade 3 flow of 29-43%, and no patients experienced 30-day death, recurrent acute MI, cardiogenic shock, stroke or anaphylaxis during the study. In a dose escalation study of a single intravenous bolus, the novel fibrinolytic agent, BB-10153 was associated with a rise in the mean area under the curve and Cmax for drug concentration over the dose range 1 to 10 mg/kg. Higher doses were associated with a range of TIMI grade 3 flow of 29-43%, and no patients experienced 30-day death, recurrent acute MI, cardiogenic shock, stroke or anaphylaxis during the study.

Rectal and parenteral paracetamol, and paracetamol in combination with NSAIDs, for postoperative analgesia

We have reviewed the analgesic efficacies of rectal and parenteral paracetamol and tested the evidence for a possible additive analgesic effect of the combination of paracetamol with a non-steroidal anti-inflammatory drug (NSAID) in postoperative pain. Randomized controlled trials were evaluated. Outcome measures were pain scores and demand for supplementary analgesia. Eight studies compared rectal paracetamol with placebo. One study of single-dose administration of rectal paracetamol 40-60 mg kg(-1) and three studies of repeat dosing with 14-20 mg kg(-1) showed significant analgesic efficacy, while studies of a single dose of 10-20 mg kg(-1) were negative. Ten studies compared parenteral paracetamol with placebo and eight studies showed improved pain relief with paracetamol. Of the nine studies comparing paracetamol with a combination of paracetamol and an NSAID, six studies showed improved pain relief for the combination while only two of the six studies comparing an NSAID with a combination of an NSAID and paracetamol showed improved pain relief for the combination. Considering the few studies available, evidence was found of a clinically relevant analgesic effect of rectal and parenteral paracetamol. Concurrent use of paracetamol and an NSAID was superior to paracetamol alone but no evidence was found of superior analgesic effect of the combination compared with the NSAID alone.

Effects of kojic acid on thyroidal functions in rats by single-dose administration and in cultured rat thyroid cells (FRTL-5 cells).

The effects of kojic acid (KA) on thyroidal function were studied by single-dose administration in rats and in cultured rat thyroid cells (FRTL-5 cells). In rats receiving a single dose of 1,000 mg/kg KA orally, the 125I uptake from blood into the thyroid gland was significantly lower than that of the control group from 30 min to 24 hr after administration. The 125I organification activity of the KA groups was significantly lower than control from 30 min to 6 hr after administration. However, the 125I organification activity at 24 hr or 48 hr after administration recovered enough to be nearly comparable with the control group. In the study in FRTL-5 cells, KA inhibited iodine organification dose-dependently, but did not inhibit iodine uptake. These results suggest that the observed lower iodine uptake activity in the single-dose administration study in rats was due to the inhibition of iodine organification caused by the oral administration of KA, consequently decreasing iodine in the entire thyroid gland. Although serum T4 showed a tendency to decrease from 2 hr to 48 hr after administration of KA, serum TSH did not show any evident change associated with KA in the single-dose administration study in rats. Based on these results, it is presumed that a massive dose or long administration period might be needed to decrease serum T4 and increase serum TSH. From these results, it is presumed that KA affected thyroidal function when given at a massive dose or in a long administration period by inhibiting iodine organification in the thyroid.

Enantioselective analysis of metoprolol in plasma using high-performance liquid chromatographic direct and indirect separations: applications in pharmacokinetics

Direct enantioselective separation on chiral stationary phases and indirect separation based on the formation of diastereomeric derivatives were developed and compared for the HPLC analysis of R(+) and S(−)-metoprolol in human plasma. Plasma samples prepared using solid-phase extraction columns or liquid–liquid extraction were directly analyzed on a Chiralpack® AD or on a Chiralcel® OD-H columns, respectively. S-(−)-menthyl choroformate was also used to yield diastereomeric derivatives resolved on a RP-8 column. The methods were employed to determine plasma concentrations of metoprolol enantiomers in a pharmacokinetic study of single dose administration of racemic metoprolol to a healthy Caucasian volunteer phenotyped as extensive metabolizer of debrisoquine. The correlation coefficients among enantioselective metoprolol plasma concentrations (5–223 ng/ml) obtained by the three methods were equal or higher than 0.99. The direct method that employed the chiral column Chiralpak® AD may be considered the most sensitive, although the three methods demonstrated interchangeable use in the pharmacokinetic investigation.

Pilot study of plasma concentrations of mosapramine, a new iminodibenzyl antipsychotic agent, after multiple oral administration in schizophrenic patients

Plasma concentrations of mosapramine, a new iminodibenzyl antipsychotic agent, were evaluated after multiple oral administration in schizophrenic patients. Mosapramine was administered orally to nine schizophrenic patients at doses of 25 mg, 50 mg, and 75 mg three times a day for about 8 weeks, and plasma concentrations of intact mosapramine were determined. Plasma concentrations could not be determined in two of the nine patients because of interference from concomitant drugs; however, the following results were obtained from seven patients. Plasma concentrations of intact mosapramine reached steady-state levels 1 to 2 weeks after the initiation of multiple-dose administration. Large interindividual differences were observed in plasma concentrations of intact mosapramine. In addition, some patients' plasma concentrations in the steady state were higher than the estimated concentrations. However, in the single-dose administration study after multiple-dose administration, no significant difference was observed in area under the curve (AUC) between estimated values and observed values. Furthermore, no significant difference was seen in elimination half-life between single-dose administration studies before and after multiple-dose administration. These results suggest that the pharmacokinetic parameters of mosapramine are not significantly varied by multiple-dose administration.