HepG2 cell viability is strongly inhibited by methazine at concentrations of 100, 300, and 1000 µM [1]. HepG2 cells' energy metabolism is promoted to switch from aerobic tricarboxylic acid cycle (TCA) and oxidative phosphorylation to anaerobic glycolysis by methazine (3, 10, 30 µM; 24 h) [1]. In HepG2 cells, promethazine (3, 10, 30 µM; 24 h) suppresses Complex IV activity [1].

There is an accumulation of triclofenac (46.3, 139, 417 mg/kg; po; single) in the kidney (18.64%), brain (23.58%), stomach (21.94%), and liver (35.84%)[1]. Promethazine (46.3, 139, 417 mg/kg; po; single) elevates MDA levels in mice's liver and brain more than in any other organ[1].

RT-PCR[1]
Cell Types: HepG2 cells
Tested Concentrations: 3, 10, 30 µM
Incubation Duration: 24 h
Experimental Results: Dramatically increased lactate dehydrogenase B (LDHB) levels when at 30 µM, and slightly increased 6-phosphofructo-2-kinase /fructose-2,6-biphosphatase 3 (PFKFB3). diminished ATP levels in a concentration-dependent manner to 91.3, 87.9 and 67.2% of the levels in the vehicle control under treatment with 3, 10 and 30 µM buprofezin, respectively. Dramatically increased the lactate levels.

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Immunofluorescence[1]
Cell Types: HepG2 cells
Tested Concentrations: 3, 10, 30 µM
Incubation Duration: 24 h
Experimental Results: Dramatically inhibited the activity of Complex IV to 82.2, 69.2 and 63.4% of the vehicle control levels following buprofezin treatment at 3, 10 and 30 µM, respectively.

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Cell Viability Assay[1]
Cell Types: HepG2 cells
Tested Concentrations: 3, 10, 30 µM
Incubation Duration: 24 h
Experimental Results: Dramatically increased the intracellular ROS levels in a concentrate-independent manner, and diminished mtDNA contents.

Animal/Disease Models: Male C57BL/6 mice (6 to 8weeks old)[1].
Doses: 46.3, 139, 417 mg/kg
Route of Administration: Oral administration; single
Experimental Results: Tended to elevate the MDA level in all organs, and the most significant concentration-dependent increases were observed in the liver and brain. demonstrated the highest concentrations in the liver (35.84%) followed by the brain (23.58%), stomach (21.94%) and kidney (18.64%), while the levels in the mouse spleen and heart were below the limit of detection.

Absorption, Distribution and Excretion
(14)C-buprofezin (radiochemical purity >97%) administered by gavage to 5 rats/sex/dose at 10 and 100 mg/kg; in males, 90-91% of dose eliminated by 48 hr (20-21% in urine, 69-71% in feces); in females, 87-89% of dose eliminated by 48 hr (13-14% in urine, 73-76% in feces); elimination faster in males during 1st 24 hr, but equalized by 48 hr; <1% of dose remained in body by 7 days; > or =30% of male dose and > or =38% of female dose recovered in bile at 24 hr; chromatography of urine, bile, and feces indicated extensive conjugation; bile cannulation of 3M/3F revealed that fecal metabolites were likely of bile origin.
[(14)C-phenyl]-buprofezin (2 or 22.5 mCi/mmol; >97% radiochemical purity) suspended in 1 mL of olive oil was administered by gavage to fasted /rats/ at 10 and 100 mg/kg (number of animals varied with the experiment); over 90% of administered dose was excreted by 48 hr at both concentrations; by 96 hr at both concentrations, 70-74% of dose excreted in feces (though a delay at the high dose relative to the low dose was noted through 24 hr), 21-25% in urine, very low amounts excreted as expired (14)C-CO2; at 10 mg/kg, 12% of the parent compound was excreted into the feces; Cmax in blood occurred at 9 hr for both doses after which concentrations declined biphasically (t1/2 =13 and 60 hr); peak levels of radiolabel occurred in tissues at 5-9 hr post dose, after which tissue levels decreased biphasically with a t1/2 of 3.5-15 hr and 15-72 hr for the 2 phases; by 96 hr tissue residue levels were low.
Male rats /were/ fed diet containing buprofezin at 200 or 1000 ppm for up to 24 weeks; 3/dose were sacrificed on days 2 and 4 and on weeks 1, 2, 4, 8, 12, 16, and 24; buprofezin levels measured by gas-liquid chromatography after extraction from blood, brain, liver, kidney, adipose tissue, and muscle; detection limit: 0.1 ppm; 200 ppm: only adipose tissue attained levels high enough to consistently detect, remaining stable between 4 days and 24 weeks at a mean concentration of 0.43-1.10 ppm; an occasional animal showed detectable levels in liver, while kidney, muscle, and brain never showed detectable levels; 1000 ppm: adipose tissue peaked at 10.53 ppm on day 4, declining to 3.40 ppm at 24 weeks; liver retained a stable concentration of 0.21-0.96 over the entire period; kidneys were near or below the detection limit for 8 weeks and undetectable thereafter; brain was near or below the detection limit for 1 week and undetectable thereafter; muscle was near or below the detection limit for 2 weeks and undetectable thereafter (no measurement at 4 weeks); test article thus did not accumulate in any tissue at either concentration.

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Metabolism / Metabolites
[(14)C-phenyl]-buprofezin (2 or 22.5 mCi/mmol; >97% radiochemical purity) suspended in 1 mL of olive oil was administered by gavage to fasted /rats/ at 10 and 100 mg/kg (number of animals varied with the experiment); ... metabolite studies revealed hydroxylation of the phenyl ring, oxidation of sulfur, and cleavage of the thiadiazin ring, with evidence of glucuronic and sulfuric conjugation.
/In/ ruminants: 2-tert-butylimino-5-(4-hydroxyphenyl)-3-isopropyl-1,3,5- thiadiazinan-4-one (BF2) was the major residue identified in liver and kidney (residue in fat and muscle were < or =0.020 ppm) and N-(4-hydroxyphenyl)acetamide (BF23) was the major residue identified in milk from the ruminant metabolism study (all other residue were <10% total radioactive residue (TRR); 3.5x maximum theoretical dietary burden (MTDB)). ...The residues of concern in milk are buprofezin and BF23; ... the residues of concern in ruminant tissues are buprofezin and BF2.

Toxicity Summary
IDENTIFICATION AND USE: Buprofezin is a solid. It is used as insecticide (chitin synthesis inhibitor). HUMAN EXPOSURE AND TOXICITY: There was no buprofezin-related increase in abnormal cells or in aberrations in human lymphocyte cultures with or without metabolic activation. ANIMAL STUDIES: The skin of rats was treated for 6 hours/day for 24 days with 0, 100, 300 or 1000 mg/kg/day. There was an increased incidence of focal hepatocellular necrosis for the 1000 mg/kg females. In long-term rat studies, increases in thyroid weight at 6, 12, and 24 month and in liver weight at 12 and 24 months were noted. Also hepatocyte necrosis and hyperplastic nodules in both sexes, interstitial pneumonia in males, interstitial heart edema and other heart effects in females were all noted at the high dose. Reduced maternal body weight and total litter resorption were noted in developmental studies. Buprofezin was not mutagenic in five Salmonella typhimurium tester strains (TA98, TA100, TA1535, TA1537, and TA1538) with or without metabolic activation. ECOTOXICITY STUDIES: A 48-hr acute exposure of buprofezin resulted in daphnid immobility at an EC(50) of 0.44 mg/L. In a 14-day chronic exposure study of buprofezin (0, 0.025, 0.05, 0.10 and 0.15 mg/L), the development and reproduction of daphnids were all significantly affected and the body length was more sensitive than other observed parameters. However, the adverse effects of buprofezin on parental daphnids can be passed on to their offspring and cannot be recovered in a short time. Malformations were observed when the embryos and larvae of African catfish (Clarias gariepinus) were exposed to more than 5 mg/L.

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Interactions
... In the present study, the combined effects of the heavy metal nickel (NiSO4) and insect growth regulator buprofezin on the induction of embryo toxicity in zebrafish were assessed. By applying nonlinear regression to the concentration-response data with each of the chemicals using the Hill and Langmuir functions and computing the predictions using the model of concentration addition (CA), we confirmed that NiSO4 and buprofezin acted together to produce synergistic embryotoxicity in zebrafish. Subsequently, we further found that the combination of NiSO4 and buprofezin formed a complex that facilitated the uptake of nickel (Ni) and buprofezin by the embryos. Following this, we clarified that an oxidative mechanism of the complex might underlie the synergistic embryotoxicity of NiSO4 and buprofezin.

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Non-Human Toxicity Values
LD50 Mouse oral >5 g/kg
LD50 Rat dermal >5 g/kg
LD50 Rat (male) oral 2198 mg/kg
LD50 Rat (female) oral 2355 mg/kg
LC50 Rat inhalation >2.2 mg/L (4 hr) /Applaud 70 DF/

[1]. Potential hepatic toxicity of buprofezin at sublethal concentrations: ROS-mediated conversion of energy metabolism. J Hazard Mater. 2016 Dec 15;320:176-186.

[2]. Inhibition of chitin biosynthesis by buprofezin analogs in relation to their activity controlling Nilaparvata lugens Stål. Pestic Biochem Physiol, 1985, 24(3): 343-347.

Buprofezin is a 2-(tert-butylimino)-5-phenyl-3-(propan-2-yl)-1,3,5-thiadiazinan-4-one in which the C=N double bond has Z configuration. It has a role as an insecticide and a member of homopteran inhibitor of chitin biosynthesis.

C16H23N3OS

305.44

305.156

69327-76-0

Buprofezin-d6;2140803-94-5

50367

White to off-white solid powder

1.18

273°C (12 torr)

104-106°C

176-178°C

1.52-1.522

4.185

0

3

3

21

408

0

PRLVTUNWOQKEAI-UHFFFAOYSA-N

InChI=1S/C16H23N3OS/c1-12(2)19-14(17-16(3,4)5)21-11-18(15(19)20)13-9-7-6-8-10-13/h6-10,12H,11H2,1-5H3

2-tert-butylimino-5-phenyl-3-propan-2-yl-1,3,5-thiadiazinan-4-one

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: 100 mg/mL (327.40 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (8.18 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (8.18 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (8.18 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)