Absorption, Distribution and Excretion
A plant metabolism study was conducted on corn with soil application of (triazolinone-3-(14)C)amicarbazone at 1.6X the max seasonal rate; application to the soil was made pre-emergence, 4 hr after planting corn seed into containers in a greenhouse. The total radioactive residues (TRRs) were 0.988, 2.369, and 0.054 ppm in corn forage, fodder, and grain, respectively, with preharvest intervals (PHIs) of 104 to 126 days. Total extractable residues were 91 to 100% TRR. Residues were characterized/identified primarily by HPLC analysis with confirmatory analysis by TLC and LC/MS. Total identified residues were 74 to 79%TRR in corn commodities. Parent amicarbazone was the major residue identified in forage and fodder, and was found in smaller amt in grain. The major residue identified in grain was the metabolite iPr-2-OH DA MKH 3586, which was also found in forage and fodder. Other identified residues were DA MKH 3586, 4-N-glu-DA MKH 3586 and iPr-2-O-glu-DA MKH 3586 in grain, forage and fodder; and tBu-iPr-2-diOH DA MKH 3586 in forage only. Four additional glucose/glucoside conjugates were identified in small amt (< or = 5% TRR) in fodder.
... A 23 field corn field trials each consisted or 3 application scenarios - a single broadcast application made preemergence, a single preplant incorporated application (both at 0.45 lb ai/acre) and an early postemergence application (0.25 lb ai/acre). All applications were made using ground application equipment. Even though the early postemergence application rate was the lowest of the 3 applications, it resulted in the highest combined residues of amicarbazone and its metabolites DA MKH 3586 and iPr-2-OH DA MKH 3586; residues following preplant incorporated application were slightly higher than those following preemergence application. Combined residues were at most 0.56 ppm in forage, 0.43 ppm in stover, and approx 0.05 ppm in field corn grain.
... (Triazolinone-3-(14)C)amicarbazone ... was admin orally to 2 lactating goats at an avg of 101 ppm in the diet (42x the max theoretical dietary burden to dairy cattle). The goats were dosed once per day for 3 consecutive days. Milk was collected twice daily throughout the study, and tissues (muscle, fat, liver, and kidney) were collected at sacrifice. Radioactive residues were highest in the meat by-products liver and kidney, followed by muscle, fat, and milk ... Total residues amounting to 91 to 99% TRR were identified in goat matrices. Both parent amicarbazone and metabolite DA amicarbazone were major residues; amicarbazone was a major residue in all matrices with the exception of kidney. Metabolite iPr-2-OH DA amicarbazone was a major residue in kidney but was a minor residue in all other matrices. Metabolite t-Bu-OH MKH 3586 was also identified as a major residue in milk, muscle, and kidney, but was a minor residue in fat and liver. Metabolite iPr-Ene DA MKH 3586 was a major residue in liver but was a minor residue in all other matrices. Other identified metabolites were found at < or = 9% TRR; these included triazolinone MKH 3586, triazolinone DA MKH 3586, tBu-Acid MKH 3586, tBu-iPr-2-diOH DA MKH 3586 (kidney and liver only), iPr-1,2-diOH DA MKH 3586, and tBu-OH DA MKH 3586.
... Encapsulated amicarbazone was admin orally to lactating cows for 30 days. Cows were dosed at 0.374, 1.238, or 4.538 ppm in the diet. Milk was sampled twice daily ... Samples of muscle, liver, kidney and fat were collected at sacrifice. Amicarbazone-related residues were below the method limit of quantitation (LOQ; <0.010 ppm) in whole milk samples collected over the course of the dosing period from the high-dose group ... Amicarbazone-related residues were highest in liver and kidney, and lower in muscle and fat. In the high dose group, residues were 1.165 to 1.193 ppm in liver, 0.080 to 0.127 ppm in kidney, 0.014 to 0.021 ppm in muscle, and < 0.010 to 0.012 ppm in fat ... Residues from the mid- and low-dose groups were 0.305 to 0.440 ppm and 0.191 to 0.235 ppm in liver, and 0.033 to 0.039 ppm and 0.017 to 0.018 ppm in kidney, respectively. Residues were below the method LOQ ... in muscle and fat samples from the mid-dose group ... The relationship between dosing level and amicarbazone-related residues appears to be linear in liver and kidney.
For more Absorption, Distribution and Excretion (Complete) data for AMICARBAZONE (7 total), please visit the HSDB record page.

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Metabolism / Metabolites
In a rat metabolism study ... (triazolinone-3 (14)C) MKH 3586 (amicarbazone ... radiochemical purity > 99%) in water was admin to 4 male Fischer rats as a single gavage dose at 5.25 to 5.99 mg/kg ... Overall recovery of the radioactive dose was 95% by 72 hr following admin. The majority of the dose was recovered in the urine within 24 hr (64% dose), indicating substantial absorption. Fecal excretion accounted for 27% dose within 24 hr post-dose. Only 2% dose was recovered in the excreta after 24 hr ... Concn of radioactivity in the selected tissues at 72 hr post-dose were low (0.004 to 0.073 ug/g). HPLC and LC-MS/MS analysis were used to identify parent and 9 metabolites in excreta from male rats ... Identified cmpd in excreta accounted for 74% of the dose ... Minor amt of parent were detected in both urine (approximately 2% dose) and feces (< 1% dose). The major metabolite in excreta was a hydroxylated, deaminated derivative of the parent, iPr-2-OH DA (34% dose), mainly found in the urine (32% dose). Other hydroxylated deaminated derivatives were also identified: tBu-OH DA (11% dose in urine and 4% dose in feces), and iPr-1,2-di-OH DA (6% dose in urine and < 1% dose in feces). The glucuronic acid conjugate of the parent, MKH 3586 GA, accounted for 11% dose, mainly in the feces (10% dose). Other metabolites (each < or = 3% dose) identified in excreta included: tBu-1,2-di-iPr-tri-OH DA; tBu-iPr-di-OH DA; iPr-1,3-diOH DA; tBu-OH-iPr-ene and DA. Based on the metabolic profile, the metabolism of MKH 3586 in male rats primarily involves deamination followed by hydroxylation with elimination in the urine. Parent also undergoes glucuronic acid conjugation and elimination in the feces.
In a rat metabolism study ... (triazolinone-3-(14)C) 4-methyl MKH 3586 (soil metabolite of amicarbazone ... radiochemical purity > 99%) in water was admin to 4 male Fischer rats as a single gavage dose at 4.86 to 5.22 mg/kg ... Overall recovery of the radioactive dose was 91% by 96 hr following admin. The majority of the dose was recovered in the urine within 12 hr (70% dose), with total urinary excretion accounted for 80% dose, indicating substantial absorption. Fecal excretion (0 to 24 hr post-dose) accounted for 8% dose. Approx 3% dose was recovered in the excreta after 24 hr ... At 96 hr post-dose, the concn of radioactivity in the tissues were low (0.002 to 0.028 ug/g), with < 0.1% dose in each tissue. HPLC, (1)H-NMR, and LC-MS/MS analysis identified 12 components in excreta ... 4 of these cmpd were present as chiral pairs. Identified cmpd accounted for 75% of the dose ... The overall accountability of the admin dose was ~ 82% ... The tested substance was not detected in the urine or feces. Hydroxylation of the soil metabolite at the isopropyl moiety formed the major metabolite, 4-Me-i-Pr-2-OH DA MKH 3586 (29% dose), which was found primarily in urine (28% dose). Further hydroxylation at the tertiary butyl group resulted in the formation of 4-Me-t-Bu-iPr-2-di-OH DA MKH 3586, which accounted for 12% dose. Alternatively, additional hydroxylation of isopropyl moiety formed 4-Me-iPr-1,2-di-OH DA MKH 3586, which constituted 9% dose in urine. Desmethylation of the parent formed DA MKH 3586 (<1% dose), which was then hydroxylated at the isopropyl moiety to form iPr-2-OH DA MKH 3586 (8% dose). The parent was also be hydroxylated at the tertiary butyl moiety to form 4-Me-tBu-OH DA MKH 3586 (9% dose), which was only detected in urine. The other minor identified metabolites occurred resulted from further oxidation, usually hydroxylation. The pathway for the metabolism of 4-methyl DA MKH 3586 in rats primarily involves a series of hydroxylation reaction. No conjugates were detected. /4-Methyl MKH 3586 (soil metabolite of amicarbazone)/
... The residues of concern in plants and livestock for tolerances and risk assessment consist of /amicarbazone ( 4-Amino-N-(1,1-dimethylethyl)-4,5-dihydro-3-(1-methylethyl)-5-oxo-1H-1,2,4-triazole-1-carboxamide)/, a desamino metabolite /(DA amicarbazone: N-(1,1-dimethylethyl)-4,5-dihydro-3-(1-methylethyl)-5-oxo-1H-1,2,4-triazone-1-carboxamide), and a hydroxylated derivative of the latter /(iPr-2-OH DA amicarbazone: N-(1,1-dimethylethyl)-4,5-dihydro-3-(1-hydroxyl-1-methylethyl)-5-oxo-1H-1,2,4-triazone-1-carboxamide). The desamino metabolite is also included in the drinking water assessment along with its N-methyl derivative ...
/It was/ proposed that the major metabolic pathway in corn involves the deamination of the triazole amino group to form the desamino metabolite (DA MKH 3586) followed by hydroxylation at the tertiary carbon of the isopropyl group to form iPr-2-OH DA MKH 3586. Additional pathways involve hydroxylation of the isopropyl methyl to form iPr-1-OH DA MKH 3586, followed by glucosidation; hydroxylation of the t-butyl and isopropyl groups to form tBu-iPr-2-diOH DA MKH 3586. In addition, DA MKH 3586 formed an N-glucoside; and glucosidation of hydroxylated DA MKH 3586 formed several minor O-glucosides.
For more Metabolism/Metabolites (Complete) data for AMICARBAZONE (10 total), please visit the HSDB record page.

Weed Science 57(6):579-583. 2009

Amicarbazone is a member of the class of triazoles that is 4,5-dihydro-1H-1,2,4-triazol-5-one which is substituted at position 1 by a tert-butylaminocarbonyl group and at position 3 by an isopropyl group. A selective herbicide for pre- and post-emergence control of annual dicotyledonous weeds and grasses, it is not approved for use within the European Union. It has a role as a herbicide. It is a member of triazoles and a carbohydrazide.

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Mechanism of Action
In a ... mechanistic subchronic oral toxicity study ... MKH 3586 (amicarbazone; > or = 98% ai ... ) was admin to 25 Fischer 344 rats/sex/group in the diet at dose levels of 0, 50, 1250, or 2500 ppm (equiv to 0/0, 0.8/0.6, 19.4/13.5, or 40.0/28.8 mg/kg/day in males/females) for 10 wk. Additionally, 5 rats/sex/group were fed test diets containing 0 and 2500 ppm for 10 wk followed by a 4-wk recovery period ... In the 1250 ppm females, TSH, free T3, T3, free T4 and T4 were incr by 27 to 61%. In the 2500 ppm females, levels of these thyroid hormones were incr over controls by 5 to 24% (not significant (NS)) but to a lesser extent than at 1250 ppm. Incr T3 (52%) and free T3 (31%; NS) were observed in males at 2500 ppm. At the end of the recovery period, levels of TSH, T3 and T4 in 2500 ppm males and females were comparable to controls. In the perchlorate assay, thyroid to blood ratio of (125)I in MKH 3586 treated groups were comparable to negative controls in males and females, indicating that the incr in thyroid hormones is not due to incr synthesis ... There were no treatment-related effects on thyroid weight or microscopic changes in the thyroid at any dose. In the liver, UDP-GT activity was incr by 109 to 241% in the > or = 1250 ppm males and females and remained incr by 27% in the 2500 ppm females at the end of recovery. Abs/relative (to body) liver weights were incr in ... 1250 males (9/14%) and females (5/12%) and in the 2500 ppm males (25/32%) and females (18/31%). Relative liver weights remained incr (7%) in the 2500 ppm males at the end of the recovery period. Abs liver weights in males and abs and relative liver weights in females were comparable to controls at the end of the recovery period. Because metabolism of the test substance is primarily via glucuronidation by UDP-GT, it is postulated that MKH 3586 competitively inhibited UDP-GT glucuronidation of T3 and T4 at 1250 ppm, resulting in incr levels of these thyroid hormones in the blood serum. At 2500 ppm, further induction of UDP-GT began to compensate for this competitive inhibition, allowing T3 and T4 levels to decline. In the females, terminal bw were decr by 7 to 10% at > or = 1250 ppm ... and remained decr by 8% at 2500 ppm at the end of recovery. An incr in the number of animals spilling food was noted at 2500 ppm. Food consumption was intermittently decr on a g/animal/day basis by 3 to 16% (p < or = 0.05) in the: (i) ... males at 2500 ppm during treatment; (ii) ... females at > or = 1250 ppm during treatment; and (iii) 2500 ppm females at the end of the recovery period. On a g/kg/day basis, food consumption was similar among groups. Abs/relative uterine weights were decr at 1250 ppm (15/9%) and 2500 ppm (24/15%). Decr in abs (20%) and relative (13%; NS) uterine weight continued in the 2500 ppm recovery group animals. There were no effects of treatment on the number of estrous cycles. However, a minor incr of 4% in the duration of the estrous cycle was observed at 2500 ppm. In conclusion, thyroid hormones were incr in the > or = 1200 ppm females and 2500 ppm males. However, thyroid to blood ratios of (125)I in treated groups were comparable to negative controls, indicated there was no impairment of thyroid hormone synthesis. Thus, the differences in thyroid hormones must be due to metabolism at an extra-thyroidal site. The liver was implicated as this site based on incr liver weights and UDP-glucuronosyltransferase activity.
... Possible interactions of MKH 3586 (amicarbazone; 98.2% ai ... ), MKH 3594 (N-desamino metabolite; 99.4% ai ... ) and KOK 9422 (putative hydrolysis metabolite; 100% ai ... ) with the enzymes involved in the synthesis of thyroid hormones and the regulation of the hypothalmus-pituitary-thyroid axis were investigated using 3 in vitro systems: (i) thyroid peroxidase (TPO), the key enzyme responsible for organification of iodine and the coupling of iodinated tyrosine residues to both T3 and T4; (ii) ioidothyronine deiodinase type I (ID-I), catalyzing the phenolic ring deiodination of T4 to form T3 in peripheral organs like thyroid, liver and kidney; and (iii) iodothyronine deiodinase type II (ID-II), catalyzing the phenolic ring deiodination of T4 to T3 in the hypothalamus and pituitary gland. Positive controls adequately demonstrated the sensitivity of each of these in vitro assays through strong, concn-dependent inhibition of the enzyme of concern ... Neither MKH 3586, MKH 3594, nor KOK 9422 inhibited TPO, ID-I or ID-II at concn up to 1mM, suggesting that MKH 3586 does not affect the iodide organification step of thyroid hormone synthesis (via either inhibition of TPO or trapping of iodine) or the peripheral metabolism of thyroid hormones via Type I or Type II deiodinases in vivo. These findings support the conclusion that MKH 3586 does not affect the major enzymes involved in the synthesis or regulation of thyroid hormones and are consistent with the findings of the ... subchronic mechanistic study in rats ...
Amicarbazone has herbicidal activity against annual broadleaf weeds. It selectively inhibits acetolactase synthase, and enzyme involved in photosystem II of plants.

C10H19N5O2

241.29016

241.153

129909-90-6

153920

Colorless crystals

1.3±0.1 g/cm3

137.5ºC

1.587

-0.99

2

4

2

17

369

0

O=C(N1N=C(C(C)C)N(N)C1=O)NC(C)(C)C

ORFPWVRKFLOQHK-UHFFFAOYSA-N

InChI=1S/C10H19N5O2/c1-6(2)7-13-15(9(17)14(7)11)8(16)12-10(3,4)5/h6H,11H2,1-5H3,(H,12,16)

4-amino-N-tert-butyl-5-oxo-3-propan-2-yl-1,2,4-triazole-1-carboxamide

BAY-314666; BAY-MKH 3586; BAY314666; BAYMKH 3586; BAY 314666; BAY MKH 3586

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)

H2O : ~1 mg/mL (~4.14 mM)

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.)