Typically sets of 4 non-fasted rats/sex were gavage-dosed daily for 14 days with Flubendiamide (NNI-0001), (14)C-labeled on the phthalic acid ring, radiochemical purity after purification of 99.2%, in an aqueous suspension of 2% sodium carboxymethylcellulose containing 0.4% Tween 80. This study assessed ADME features including quantification of major metabolites. Rats were sacrificed either 9 hr, 24 hr, or 168 hr after the final (14th) dosing. ... Females showed less metabolized label and greater longterm tissue retention of label than males, consistent with other studies. Parent flubendiamide comprised 82% and 91% of fecal metabolites in males and females, respectively. Relative proportions of remaining fecal label consisted of the benzyl alcohol metabolite (A-16) (7.2% of excreted radioactivity in males, and 2.2% in females), the benzoic acid metabolite (A-18) (2.8% of excreted radioactivity in males and below detection in females), with other metabolites uncharacterized or in very small amounts. This study reported quantifiable amounts of the iodophthalimide metabolite (A-14) of flubendiamide in fat. This metabolite, involving cleavage of all components distal to the phthalamide nitrogen, had not been found in measurable amounts in excreta.
Typically sets of 4 fasted rats/sex were dosed once by gavage with Flubendiamide (NNI-0001), (14)C-labeled on either the phthalic acid ring or (to a limited extent) on the aniline ring, radiochemical content after purification > 99.5%, in an aqueous suspension of 1% sodium carboxymethylcellulose containing 0.2% Tween 80. This study assessed ADME features including identification of major metabolites following treatment with either 2 or 200 mg/kg flubendiamide. There was no significant excretion of label into expired air. Over 90% of administered dose was excreted in feces regardless of dose or sex. Although urine was a minor route (< 2% of administered dose), males consistently passed more label in urine than did females. High dose levels led to markedly reduced absorption. There was no obvious difference in excretion patterns and there were minimal differences in metabolite chromatograms between the two labeled forms, indicating that there was very little cleavage dividing major portions of the molecule. Tmax was 6 to 12 hours for blood and plasma. Tissue distribution of label was preferentially to the g.i. tract, liver and other highly perfused organs, and body fat. Low dose treatments found peak tissue levels in males several-fold higher than females. Tissue levels in males were lower by about 90% from 9 hrs to 24 hrs, and by another 90% from 24 hrs to 168 hrs. Females did not clear tissues as rapidly as males, however, so females had higher residual tissue concentrations at 168 hours than males. The primary substituent available for oxidation on flubendiamide is the methyl group on the aniline ring. The most common products were the benzyl alcohol (designated A-16) and the benzoic acid (designated A-18) of that methyl carbon. The less abundant intermediate benzaldehyde was designated as A-17. Substantial sex differences emerged in metabolic profiles. In the feces, the benzyl alcohol was the dominant metabolite in 2 mg/kg males (31-37% of administered dose), followed by parent flubendiamide (15-30% of administered dose), and the benzoic acid metabolite (15-16% of administered dose). In feces of 2 mg/kg females, 66% of administered dose was flubendiamide, 5-6% of administered dose was benzyl alcohol, and there was very little benzoic acid metabolite. There were no other major metabolites. Parent flubendiamide was the predominant labeled constituent in feces of both sexes at 200 mg/kg, indicating saturable absorption. Female rats were very slow to clear radioactivity: contrast liver label content in 2 mg/kg males dropping from 5.628% to 0.104% of administered dose at 9 hrs and 168 hrs after dosing vs. content in females of 1.623% and 1.305% of administered dose at the same sampling times. Female rats have limited hepatic metabolic capacity for flubendiamide, hence there was appreciable accumulation of parent flubendiamide in the livers of females at 24 hours (an order of magnitude more than all extractable metabolites combined). In males, very little parent flubendiamide was found in liver (less than half of the percent of dose represented by benzyl alcohol or benzaldehyde metabolites). Liver in males had smaller but measurable amounts of the sole identified product of cleavage between the two ring structures of flubendiamide (the aniline fragment).
Flubendiamide (purified to 99.6% purity for non-labeled a.i. and for (14)C-labeled on the phthalic acid ring) was administered once by gavage (aqueous suspension of 2% sodium carboxymethylcellulose containing 0.4% Tween 80) to F-344 (F344/DuCrj) rats provided with biliary cannulae. Three males and 6 females received 2 mg/kg in the definitive study. (Additional rats were dosed twice at 16-hr intervals with 20 mg/kg a.i. to facilitate characterization of metabolites.) ... The only quantifiable metabolite observed in feces or gastrointestinal content was the benzyl alcohol metabolite, possibly resulting from intestinal microbial action of non-absorbed material. Much of the biliary profile in males consisted of oxidation products of the methyl group on the aniline ring ... . There was also some oxidation of the methyl groups between the phthalamide nitrogen and the sulfonyl group (including compounds designated A-19, A-20, A-25, and A-29 by investigators). Most commonly the oxidation products of the methyl group on the aniline ring were not conjugated, however it appears that oxidation of a methyl group between the phthalamide nitrogen and the sulfonyl group frequently was followed by glucuronide conjugation in males. Both sexes were capable of glutathione formation of parent compound, conjugation proposed by investigators to be on the phthalic ring. In females, glutathione conjugates (and downstream derivatives of the conjugation) were the dominant biliary metabolites. A cyclic acetal was observed in bile, particularly in males.
Rat, dog, mouse, and human microsomes (from both sexes in all species) were all able to metabolize flubendiamide to A-16, except for microsomes from female rats. (Human microsomes, but not rat microsomes, additionally produced small amounts of hydroxybenzoic acid metabolite, A-20, however metabolism to A-16 appears to be a valid indicator of overall flubendiamide metabolism.) Antisera to CYP isoforms tested in male rat microsomes found that anti-CYP3A2 sera had no effect on A-16 production, whereas anti-CYP2C11 markedly inhibited A-16 production. In contrast, recombinant microsomes expressing rat CYP3A2 produced A-16, whereas other isoforms including CYP2C11 recombinant microsomes did not. Antibody effects on human liver microsome activities found that anti-rat CYP3A2 and anti-human CYP3A4 caused similar and substantial inhibition in microsomes from male or female human liver (it was noted that anti-rat CYP3A2 was strongly reactive toward CYP3A4). Of 5 recombinant microsomes expressing human liver P450 isoforms evaluated for A-16 production, only CYP3A4 yielded measurable A-16.
F-344 (F344/DuCrj) rats and Crlj: CD1 (ICR) mice (4/species/sex/dosing period) were dosed with 200 mg/kg/day unlabeled flubendiamide, purity 96.7% by gavage in corn oil. Treatments were daily for 1, 7, or 14 days. Animals were sacrificed 24 hr after the final dose. ...Concentrations of flubendiamide and of the iodophthalimide metabolite (A-14) /were determined/ in plasma, liver, and fat. Flubendiamide did not increase in these tissues over time in males. There was a marginal accumulation in females in all studied tissues, reaching steady state by day 7. A-14 appeared to be accumulating in fat in rats of both sexes, reaching steady state by day 7. Flubendiamide was always more concentrated in liver and fat than in plasma, and concentrations were much higher in females than in males. Day 7 concentrations of flubendiamide in liver and fat of male rats at day 7 were 1.3 and 8.9 mg/kg, respectively. Liver and fat concentrations in female rats were 26.7 and 68.0 mg/kg. A-14 concentrations in male and female rats for liver and fat at day 7 were identical at < 1.0 and 2.9 mg/kg, respectively. Flubendiamide concentrations on day 7 in liver and fat of male mice were 2.3 and 3.4 mg/kg, respectively. Corresponding levels in female mice were 2.4 and 1.9 mg/kg (i.e. much unlike the marked accumulation in female rats). A-14 levels were generally below quantification limits in both male and female mice. This study indicates that limited metabolic capacity of female rats toward flubendiamide and tendency to metabolize significant amounts of A-14 do not apply to either sex of mice.
/SRP:/ Immediate first aid: Ensure that adequate decontamination has been carried out. If patient is not breathing, start artificial respiration, preferably with a demand valve resuscitator, bag-valve-mask device, or pocket mask, as trained. Perform CPR if necessary. Immediately flush contaminated eyes with gently flowing water. Do not induce vomiting. If vomiting occurs, lean patient forward or place on the left side (head-down position, if possible) to maintain an open airway and prevent aspiration. Keep patient quiet and maintain normal body temperature. Obtain medical attention. /Poisons A and B/
/SRP:/ Basic treatment: Establish a patent airway (oropharyngeal or nasopharyngeal airway, if needed). Suction if necessary. Watch for signs of respiratory insufficiency and assist ventilations if needed. Administer oxygen by nonrebreather mask at 10 to 15 L/min. Monitor for pulmonary edema and treat if necessary ... . Monitor for shock and treat if necessary ... . Anticipate seizures and treat if necessary ... . For eye contamination, flush eyes immediately with water. Irrigate each eye continuously with 0.9% saline (NS) during transport ... . Do not use emetics. For ingestion, rinse mouth and administer 5 mL/kg up to 200 mL of water for dilution if the patient can swallow, has a strong gag reflex, and does not drool ... . Cover skin burns with dry sterile dressings after decontamination ... . /Poisons A and B/