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-ibuprofen (radiochemical purity >97%) was administered by gavage to 5 rats/sex/dose at doses of 10 and 100 mg/kg, respectively. Male rats eliminated 90-91% of the dose within 48 hours (20-21% excreted in urine and 69-71% in feces); female rats eliminated 87-89% of the dose within 48 hours (13-14% excreted in urine and 73-76% in feces); male rats eliminated the dose faster in the first 24 hours, but the elimination rate became more uniform after 48 hours; the residual dose in the body was <1% after 7 days; after 24 hours, the recovered dose in the bile of male rats was ≥30%, and the recovered dose in the bile of female rats was ≥38%; chromatographic analysis of urine, bile and feces showed extensive binding reactions; bile cannulation analysis of 3M/3F showed that fecal metabolites may be derived from bile.
[(14)C-phenyl]-ibuprofenazine (2 or 22.5 mCi/mmol; radiochemical purity >97%) was suspended in 1 mL of olive oil and administered to fasted rats by gavage at doses of 10 and 100 mg/kg (number of animals varied with the experiment); at both concentrations, more than 90% of the administered dose was excreted within 48 hours; after 96 hours, 70-74% of the dose was excreted in feces at both concentrations (although there was a delay of 24 hours in the high-dose group compared to the low-dose group), 21-25% was excreted in urine, and a very small amount was excreted as exhaled (14)C-CO2; at a dose of 10 mg/kg, 12% of the parent compound was excreted in feces; at both doses, the peak concentration of the drug in the blood occurred 9 hours after administration, followed by a biphasic decline (half-lives of 13 hours and 60 hours, respectively); the peak level of the radiolabeled substance in tissues occurred 5-9 hours after administration. After 96 hours, tissue levels showed a biphasic decline, with half-lives of 3.5–15 hours and 15–72 hours for the two phases, respectively; after 96 hours, tissue residue levels were low. Male rats were fed diets containing 200 or 1000 ppm ibuprofen for up to 24 weeks. Three rats from each dose group were sacrificed on days 2 and 4, and at weeks 1, 2, 4, 8, 12, 16, and 24. Ibuprofen was extracted from blood, brain, liver, kidney, adipose tissue, and muscle, and its concentration was determined by gas chromatography. The detection limit was 0.1 ppm. At 200 ppm, only adipose tissue reached a sufficiently detectable concentration, remaining stable from day 4 to week 24, with an average concentration of 0.43–1.10 ppm. Concentrations were detected in the liver of some animals, but never in the kidney, muscle, or brain. At 1000 ppm, adipose tissue peaked at 10.53 ppm on day 4, decreasing to 3.40 ppm at week 24. The liver maintained a stable concentration of 0.21–0.96 ppm throughout the period. The kidney showed a stable concentration of 0.21–0.96 ppm at day 8. The levels were close to or below the detection limit within one week and then became undetectable; in the brain, the levels were close to or below the detection limit within one week and then became undetectable; in the muscle, the levels were close to or below the detection limit within two weeks and then became undetectable (no measurements were taken at four weeks). Therefore, the test substance did not accumulate in any tissue at either concentration.

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

Toxicity Summary
Identification and Uses: Thiazide is a solid. It is used as an insecticide (chitin synthesis inhibitor). Human Exposure and Toxicity: No abnormal cells or increases associated with thiamethoxam were observed in metabolically activated or inactive human lymphocyte cultures. Animal Studies: Rat skin was treated daily with 0, 100, 300, or 1000 mg/kg/day of thiamethoxam for 6 hours for 24 days. Female rats in the 1000 mg/kg dose group showed an increased incidence of focal hepatocellular necrosis. In long-term rat studies, increased thyroid weight and liver weight were observed at 6, 12, and 24 months. Furthermore, high-dose groups also showed hepatocellular necrosis and proliferative nodules in both sexes, interstitial pneumonia in males, interstitial cardiac edema in females, and other cardiac damage. Developmental Studies: Maternal weight loss and complete offspring reabsorption were observed. Regardless of metabolic activation, ibuprofen was not mutagenic in any of the five tested Salmonella typhimurium strains (TA98, TA100, TA1535, TA1537, and TA1538). Ecotoxicity studies: 48 hours after acute ibuprofen exposure, Daphnia davidii exhibited activity impairment at an EC50 of 0.44 mg/L. In a 14-day chronic exposure study to ibuprofen (0, 0.025, 0.05, 0.10, and 0.15 mg/L), development and reproduction of Daphnia davidii were significantly affected, with body length being more sensitive than other observed parameters. However, the adverse effects of ibuprofen on parental Daphnia davidii were passed on to their offspring and did not recover in the short term. Malformations were observed in African catfish (Clarias gariepinus) embryos and larvae exposed to ibuprofen concentrations exceeding 5 mg/L.

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Interaction
…In this study, we evaluated the synergistic effect of the heavy metal nickel (NiSO4) and the insect growth regulator ibuprofen on zebrafish embryotoxicity. By performing nonlinear regression on the concentration-effect data of each chemical using Hill and Langmuir functions, and calculating predicted values using a concentration summation (CA) model, we confirmed that the combined effects of NiSO4 and thiamethoxam have synergistic toxicity on zebrafish embryos. Subsequently, we further discovered that NiSO4 and thiamethoxam form a complex that promotes embryonic uptake of both nickel (Ni) and thiamethoxam. We then elucidated that the oxidation mechanism of this complex may be a potential mechanism for the synergistic embryotoxicity of NiSO4 and buprenorphine.

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Non-human toxicity values
Oral LD50 in mice >5 g/kg
Dermal LD50 in rats >5 g/kg
Oral LD50 in male rats 2198 mg/kg
Oral LD50 in female rats 2355 mg/kg
Inhalation LC50 in rats >2.2 mg/L (4 hours) /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-(propyl-2-yl)-1,3,5-thiadiazinane-4-one, in which the C=N double bond has a Z configuration. It is an insecticide and also one of the inhibitors of chitin biosynthesis in homoptera insects.

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

CC(C)N1C(=NC(C)(C)C)SCN(C1=O)C2=CC=CC=C2

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.)