4-hydroxyphenylpyruvate dioxygenase (HPPD); Carotenoid biosynthesis

Pyrasulfotole is a pyrazole pesticide that is 1,3-dimethylpyrazol-5-ol in thich the hydrogen at position 4 has been replaced by a 2-(methylsulfonyl)-4-(trifluoromethyl)benzoyl group. A 4-hydroxyphenylpyruvate dioxygenase inhibitor, it is used as a herbicide (particularly in conjunction with the safener mefenpyr-diethyl) to control various broad-leaved weeds. It has a role as a herbicide, a carotenoid biosynthesis inhibitor, an EC 1.13.11.27 (4-hydroxyphenylpyruvate dioxygenase) inhibitor and an agrochemical. It is a pyrazole pesticide, an aromatic ketone, a sulfone and a member of (trifluoromethyl)benzenes [1].

Absorption, Distribution and Excretion [1]
Following oral administration of 10 mg/kg phenyl or pyrazole ring-labeled pyrasulfotole, approximately 60% of radiolabeled compound was excreted in the urine after 6 hours, while approximately 73% of the administered dose was recovered in the urine by the time of sacrifice (52 hours). Therefore, approximately 60% of the compound was absorbed within 6 hours of exposure. Less than 2% of the administered dose remained in the residual carcass and tissues at sacrifice, and the highest residues were found in the liver and kidney. Approximately 35% of labeled compound was excreted in the feces 52 hr after dosing, approximately 25% of was parent.

In a metabolism study, AE 0317309 (/pyrasulfotole/ 97.6-100% ai, Vial #s C-938, C-939, C- 938B, C-1024A, K-1196, K-1267), male Wistar Hanover rats (five rats for each radiolabel) were given single oral doses of (phenyl-UL-(14)C) AE 0317309 or pyrazole-3-(14)C AE 0317309 at dose rates of 10.0 and 9.88 mg/kg bw, respectively. In two separate experiments, male Wistar Hanover rats with surgically implanted jugular cannula were dosed intravenously with (phenyl-UL-(14)C) AE 0317309 (four rats) or (pyrazole-3-(14)C) AE 0317309 (five rats) at dose rates of 9.81 and 9.60 mg/kg bw, respectively. The (phenyl-UL-(14)C) AE 0317309 was readily absorbed following oral dosing, with approximately 62% of the administered radioactivity dose being recovered in the urine within 6 h post dose. A total of approximately 73% of the administered radioactivity dose was recovered in the urine by the time of sacrifice (52 hr). The (pyrazole-3-(14)C) AE 0317309 was readily absorbed following a single oral dose, with approximately 57% of the administered radioactivity dose being recovered in the urine within 6 h post dose. A total of approximately 75% of the dose was recovered in the urine by the time of sacrifice (48 hr). In the case of intravenous administration, approximately 90% of the radioactivity was excreted in the urine and approximately 10% in the feces by 48 h. Most (over 80%) of the intravenous dose was excreted in the urine within 6 h. In all experiments, <2% of the administered dose remained in the residual carcass and tissues at sacrifice. In all experiments, the highest residues were found in the liver and kidney. There was no apparent difference between the phenyl-UL-(14)C label and the pyrazole-3-(14)C label in the distribution of equivalents in the tissues following intravenous dosing. However, concentrations in the tissues were consistently higher in the case of the pyrazole-3-(14)C label than in the case of the phenyl-UL-(14)C label following oral dosing. The distribution of radioactivity in tissues did not differ greatly between oral and intravenous dosing, with the exception that the concentration in the residual carcass is somewhat higher in the latter case. Following both oral and intravenous administration, most of the dose was excreted unchanged as AE 0317309. ...

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Metabolism / Metabolites
In a metabolism study, AE 0317309 (/pyrasulfotole/ 97.6-100% ai, Vial #s C-938, C-939, C- 938B, C-1024A, K-1196, K-1267), male Wistar Hanover rats (five rats for each radiolabel) were given single oral doses of (phenyl-UL-(14)C) AE 0317309 or pyrazole-3-(14)C AE 0317309 at dose rates of 10.0 and 9.88 mg/kg bw, respectively. In two separate experiments, male Wistar Hanover rats with surgically implanted jugular cannula were dosed intravenously with (phenyl-UL-(14)C) AE 0317309 (four rats) or (pyrazole-3-(14)C) AE 0317309 (five rats) at dose rates of 9.81 and 9.60 mg/kg bw, respectively. ... Following both oral and intravenous administration, most of the dose was excreted unchanged as AE 0317309. Hydroxymethyl AE 0317309, desmethyl AE 0317309, and AE B197555 were observed as minor metabolites in the urine and feces. Following oral dosing, approximately 70% of the radioactivity was excreted in the urine and 30% in the feces by 48 or 52 hr.

Antidote and Emergency Treatment
/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/

/SRP:/ Advanced treatment: Consider orotracheal or nasotracheal intubation for airway control in the patient who is unconscious, has severe pulmonary edema, or is in severe respiratory distress. Positive-pressure ventilation techniques with a bag valve mask device may be beneficial. Consider drug therapy for pulmonary edema ... . Consider administering a beta agonist such as albuterol for severe bronchospasm ... . Monitor cardiac rhythm and treat arrhythmias as necessary ... . Start IV administration of D5W /SRP: "To keep open", minimal flow rate/. Use 0.9% saline (NS) or lactated Ringer's if signs of hypovolemia are present. For hypotension with signs of hypovolemia, administer fluid cautiously. Watch for signs of fluid overload ... . Treat seizures with diazepam or lorazepam ... . Use proparacaine hydrochloride to assist eye irrigation ... . /Poisons A and B/

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Non-Human Toxicity Excerpts
/LABORATORY ANIMALS: Subchronic or Prechronic Exposure/ In a 28-day dermal toxicity study, AE 0317309 (/pyrasulfotole/ 96.3% w/w ai, batch Op:1-4) was applied to the shaved skin of 10 Wistar HsdCpb:WU rats/sex/dose at dose levels of 0, 10, 100, or 1000 mg/kg bw/day, 6 hours/day for 5 days/week during a 28-day period. There were no compound-related effects on mortality, clinical signs (including ophthalmology), body weight, food and water consumption, hematology, or gross pathology. Statistically significant increases in cholesterol (27%), triglycerides (46%), and protein (4%) were observed in males, and total bilirubin (17%) in females at 1000 mg/kg bw/day. Since the observed changes were minor and within the limits of the submitted historical control data, they were not considered toxicologically significant. Absolute liver weights were statistically significantly increased in males (13.9%) and females (11.3%) at 1000 mg/kg bw/day. Hepatocellular hypertrophy and hypertrophy of the pars distalis of the pituitary gland were observed in males at 1000 mg/kg bw/day. Focal degeneration of the pancreas was observed in 2/10 males and 1/10 females at 100 mg/kg bw/day and in 8/10 males and 8/10 females at 1000 mg/kg bw/day. An increased incidence of thyroid follicular cell hypertrophy was seen in high-dose males. Alteration of thyroid colloid was seen with increasing incidence and severity at greater than or equal to 10 mg/kg bw/day in males. However, since the alteration was predominantly minimal at 10 mg/kg, it was not considered toxicologically relevant at this dose. The LOAEL is 100 mg/kg/day, based on focal degeneration of the pancreas (both sexes) and alteration of thyroid colloid (males). The NOAEL was 10 mg/kg/day.

/LABORATORY ANIMALS: Subchronic or Prechronic Exposure/ In a 90-day oral toxicity study, AE 0317309 (/pyrasulfotole/ 95.7% w/w ai, batch Op:1-4) was administered in the diet to groups of 4 Beagle dogs/sex/dose at dose levels of 0, 100, 500, or 1000 ppm (equivalent to 0/0, 3/3, 17/17 and 40/33 mg/kg bw/day in male/female dogs, respectively) for a period of 90 days. There were no compound-related effects on mortality, clinical signs, body weight, food and water consumption, hematology, clinical chemistry, urinalysis, gross pathology and microscopic pathology. The LOAEL was not established. The NOAEL is 40 mg/kg bw/day for males and 33 mg/kg bw/day for females.

/LABORATORY ANIMALS: Subchronic or Prechronic Exposure/ In a 90-day oral toxicity study, AE 0317309 (/pyrasulfotole/ 97.4% w/w ai, batch H2235) was administered in the diet to groups of 10 Rj:WI(IOPS HAN) Wistar rats rats/sex/dose at dose levels of 0, 2, 30, 1000, 7000 and 12,000 ppm (equivalent to 0.0, 0.13, 1.96, 66, 454, and 830 mg/kg bw/day for males and 0.0, 0.15, 2.32, 77, 537 and 956 mg/kg bw/day in females) for a period of 90 days. No abnormalities were detected during the neurotoxicity assessment and there were no treatment related effects on hematological parameters. At 12000 ppm, six male and one female were either sacrificed for humane reasons or were found dead during the treatment period. The group was terminated at week 11 due to excessive toxicity. At 7000 ppm, two males were found dead or were sacrificed for humane reasons during the treatment period. Treatment related clinical signs were observed in a large number of rats at 7000 and 12,000 ppm and consisted of intensely yellow colored urine associated on a few occasions with soiled anogenital area, soiled fur, piloerection, general pallor and wasted appearance. Other clinical signs noted included: few or no feces, cold to touch, reduced motor activity, labored respiration, hunched posture, increased salivation and soiling around the mouth. White areas on eyes were noted in two males at 7000 ppm and in one male and four females at 12,000 ppm. At 1000 ppm, yellow colored urine was noted in all males on a few days and one female presented a white area on the eyes. At 12,000 ppm, a reduction in mean body weight gain of 70% was recorded in males during the first week of exposure. At this dose level, the depressions in body weight gain ranged from 11.5 to 56% over days 22 to 70. In females at 7000 ppm and 12,000 ppm, the depressions in mean body weight gain were 12.5 and 15.6%, respectively, relative to controls at the end of the 90 day period. At 12,000 ppm food consumption in males was lower than control values throughout the study. In females at 12,000, the mean food consumption was lower than control value on the first week of treatment only (reduction of 28%) without reaching statistical significance. At 7000 ppm, a reduction of food consumption was noted during the first week in males (28%) and females (15%), the difference with controls reaching statistical significance in males only. Very slight reductions thereafter were also observed in both sexes but were not statistically significant. Neovascularization of the cornea and characteristic "snowflake" corneal opacities were noted at 7000 and 12000 ppm in males, and at 1000, 7000, and 12000 ppm in females. In males, bilirubin, AST and ALT, urea, and creatinine were increased (but not statistically significantly) at 7000 ppm. Cholesterol was statistically significantly (P less than or equal to 0.01) increased in males at 1000 ppm (45%) and 7000 ppm (51%). Triglycerides were statistically significantly (P less than or equal to 0.01) increased in males at 1000 ppm (112%) and 7000 ppm (68%). Ketone levels were increased from 1000 ppm in both males and females. ... There was an increased incidence of occult blood, erythrocytes, leukocytes, and epithelial cells in the urine in both males and females at 7000 ppm and in females at 12,000 ppm (males in the 12,000 ppm group did not survive until the end of the study and urine was therefore not collected). At 1000 ppm and 7000 ppm, the relative liver to body weight in males was statistically significantly increased by 22 and 26% respectively. At 1000 ppm and 7000 ppm, relative kidney to body weight in males was increased 3.5 and 38.6%, respectively, the latter increase being statistically significant. For females relative liver weight was increased 8.7, 13 and 8.7% at 1000, 7000 and 12,000 ppm and relative kidney weight was increased 8, 25.4 and 30%, respectively. At 7000 ppm and 12,000 ppm, abnormal shape of the kidneys, mottled kidneys, dilation of and gritty content in the renal pelvis, gritty content, distension of the urinary bladder, and gritty content of the ureters, and enlarged livers were observed in males and females. Livers were enlarged in 3/10 males at 1000 ppm. Prominent lobulation was noted in 1/7 male at 7000 ppm and in 2/10 males at 1000 ppm. The thyroid gland was enlarged in one male at 1000 ppm. The 12,000 ppm group was terminated early at 11 weeks and tissues from animals in this group were not microscopically examined. Histological changes associated with the presence of calculi (urolithiasis) were found in the kidneys/urinary bladder/ureters in 4/8 males and 6/10 females at 7000 ppm. Associated histological changes included: pelvic dilatation (unilateral or bilateral), urinary epithelial hyperplasia (pelvis, urinary bladder and ureters), interstitial fibrosis of the urinary tract, cystitis and Ureteritis. Slight to moderate diffuse centrilobular hepatocellular hypertrophy was observed in 6/7 males at 7000 ppm, in 9/10 males at 1000 ppm and in 1/10 female at 7000 ppm. In females, a periportal vacuolation was found in 8/10 animals at 7000 ppm and 3/10 animals at 1000 ppm. The LOAEL is 1000 ppm (77 mg/kg bw/day) in females based on neovascularization of the cornea and "snowflake" corneal opacity; and 7000 ppm (454 mg/kg bw/day) in males based on mortality, neovascularization of the cornea and "snowflake" corneal opacity, and histopathology in the kidney, urinary bladder, thyroid, and ureters. The NOAEL is 30 ppm (2.32 mg/kg bw/day) in females and 1000 ppm (66 mg/kg bw/day) in males.

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Non-Human Toxicity Values
LD50 Rat dermal > 2,000 mg/kg
LD50 Rat oral > 2,000 mg/kg
LC50 Rat inhalation > 5.03 mg/L/4 hr

[1]. https://pubchem.ncbi.nlm.nih.gov/compound/11693711

Enzymes, coenzymes and metabolites are the reagents characteristic of enzymatic analysis. Some of these materials must be prepared or isolated in the laboratory, but many are available commercially. Information about these materials is less readily available to the analyst than information about inorganic reagents. This situation is made worse by the multiplicity and variety of the definitions of purity. In 1955, the Committee on Biological Chemistry and Chemical Technology, National Academy of Sciences, National Research Council (USA) began to collect standard values for biochemical compounds1). As a result of this work the first volume of “Specifications and Criteria of Biochemical Compounds” was published in June 1960*). This volume does not contain chapters on “Phosphate Esters” and “Enzymes”. This chapter examines the activity and the stability of biochemical reagents. The best criterion for the purity of an enzyme is the specific activity together with details of the activity of contaminating enzymes. The activity of these contaminants is expressed as a percentage of the specific activity of the enzyme described. The activities of the enzymes are given in units and the specific activity in units per mg protein. To conform to the proposed International Unit, a unit of enzyme activity is defined as the amount of enzyme which transforms 1 μmole of substrate in 1 min at 25°C. ADH contains up to 40 SH groups and 5 zinc atoms per molecule and therefore, it is sensitive to oxidizing and chelating agents. The crystalline suspension in 2.0 M (NH4)2SO4 solution containing 3% Na4P2O7 and 1% glycine is stable for several months at 0°C to 4°C when air is excluded, while at room temperature it is only stable for 1 week. Solutions or suspensions can be dialyzed against distilled water at 0°C to 3°C without any significant loss of activity. Triosephosphate isomerase is used as the auxiliary enzyme and glycerol-1-phosphate dehydrogenase as the indicator enzyme. [1]

C14H13F3N2O4S

362.32

365400-11-9

Solid Powder

Cc1c(c(n(n1)C)O)C(=O)c2ccc(cc2S(=O)(=O)C)C(F)(F)F

Sulfonylpyraclostrobin; Pyrasulfotole; 365400-11-9; Pyrasulfotole [ISO]; (5-hydroxy-1,3-dimethyl-1H-pyrazol-4-yl)(2-(methylsulfonyl)-4-(trifluoromethyl)phenyl)methanone; PYRASULFATOLE; HSDB 7879; M31E7U368M; 2,5-dimethyl-4-[2-methylsulfonyl-4-(trifluoromethyl)benzoyl]-1H-pyrazol-3-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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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