Absorption, Distribution and Excretion
Sprague-Dawley Crl:CD/BR rats (half male and half female) were administered 10 or 1000 mg/kg of [(14)C]…/biphenylhydrazine/… (radiochemical purity: >98%, specific activity: 34.4 mCi/mmol, labeled on the aromatic ring; biphenylhydrazine/… (unlabeled purity: 99.6%)) by gavage. In distribution, metabolism, and excretion studies, five animals per sex in each group were administered the drug, and urine and feces were collected for 7 days. In biliary studies, three cannulated animals per sex in each group were administered the drug, and urine, feces, and bile were collected for 72 hours. In preliminary pharmacokinetic and pharmacokinetic studies, three animals per sex in each group and five animals per sex in each group were administered the drug separately. In the preliminary studies, urine and feces were collected for 4 days, and blood samples were collected for 72 hours via jugular vein cannulation. In the primary study, urine and blood were collected from animals in the 10 mg/kg treatment group for 4 days. In the 1000 mg/kg treatment group, urine, feces, and blood were collected for 7 days. In the tissue distribution study, 9 animals per sex in each group were administered the drug. In the 10 mg/kg treatment group, 3 animals per sex were euthanized at 6, 24, and 48 hours post-administration. In the 1000 mg/kg treatment group, 3 animals per sex were euthanized at 18, 42, and 72 hours post-administration. Urine and feces were collected at specified time intervals. Tissue samples from the distribution, metabolism, and excretion studies, as well as the tissue distribution study, were processed to detect the presence of…radiolabeled substances. Radiolabeled substances were isolated from urine and feces obtained from the distribution, metabolism, and excretion studies, as well as the bile studies, and their structural analysis was performed to determine their metabolic profile. The primary route of excretion was feces in both dosage groups. For the 10 mg/kg group, 66% of the administered dose (AD) was recovered in feces, with 75-82% excreted within the first 24 hours. After 7 days, radiolabeled AD was recovered in urine and cage flushing fluid, accounting for 27-29% of the AD. For the 1000 mg/kg treatment group, 82% of AD was recovered in feces within 7 days of administration, with 46-57% recovered within the first 24 hours. The percentage of AD separated in urine and cage flushing fluid was 10-15%. Bile studies showed that bile was an important excretion route for the 10 mg/kg treatment group. Within 72 hours of administration, 69-74% of the administered dose was recovered in bile in the low-dose group. In contrast, only 21-26% of the dose was separated from bile within 72 hours in the high-dose group. Fecal recovery rates were only 7-8% in the 10 mg/kg group, compared to 56-64% in the 1000 mg/kg group. A significant portion of the high-dose dose remained unabsorbed. The following pharmacokinetic parameters were observed: tmax in males and females of the 10 mg/kg and 1000 mg/kg groups were 5 hours and 6 hours, and 18-24 hours, respectively; Cmax in males and females of the 10 mg/kg and 1000 mg/kg groups were 6.4 and 5.6 μg equivalents/g, and 119 and 71 μg equivalents/g, respectively. The corresponding t1/2 values for the 1000 mg/kg group were 11.5 hours and 13.3 hours for males of the low-dose group, and 12 hours and 15.6 hours for females of the high-dose group. In tissue distribution studies, the 10 mg/kg group reached maximum residual levels 6 hours after administration, and no radiolabeled substances were detected in any tissues. In the 1000 mg/kg group, maximum residual levels were reached in most tissues in male animals at 18 hours post-administration and in female animals at 42 hours post-administration. Significant radiomarkers were still detectable in some organs (e.g., spleen, erythrocytes, liver, and kidneys) up to 7 days post-administration. Analysis of the radiomarkers recovered from feces revealed various modifications to their parent compounds, including hydrazine oxidation, demethylation, cyclization, hydroxylation, separation into biphenyl and hydrazine carboxylic acid moieties, and conjugation with glucuronic acid or sulfuric acid. In the 10 mg/kg group, the identified fraction extracted from feces accounted for 39% of the total absorption. The major compounds were…biphenylhydrazine glucuronide (6.3–8.9% of total absorption),…biphenylhydrazine ester (4.8–7.2% of total absorption), and…4-hydroxybiphenyl (5.5–7.1% of total absorption). In contrast, in the 1000 mg/kg group, the parent compound recovered from feces accounted for 48-61% of the total absorption. Biphenylhydrazine glucuronide accounted for 4.7-5.6% of the total absorption. Overall, the test substance was well absorbed at low doses and was metabolized and bound before excretion via bile. At high doses, the dose percentage decreased significantly.

---

Metabolism/Metabolites
Sprague-Dawley Crl:CD/BR rats (both male and female) were administered 10 or 1000 mg/kg of [(14)C].../biphenylhydrazine/ (radiochemical purity: >98%, specific activity: 34.4 mCi/mmol, labeled on the aromatic ring,.../biphenylhydrazine/ (unlabeled...purity: 99.6%)). ... Analysis of the radiolabeled fraction recovered from feces revealed various modifications to the parent compound. Hydrazine is oxidized, demethylated, cyclohydroxylated, separated into biphenyl and hydrazine carboxylic acid moieties, and conjugated with glucuronic acid or sulfate. For the 10 mg/kg group, the identified fraction extracted from feces accounted for 39% of the administered dose (AD). The major compounds were.../biphenylhydrazine/glucuronide (6.3–8.9% of total intake), biphenylhydrazine (4.8–7.2% of total intake), and 4-hydroxybiphenyl (5.5–7.1% of total intake). In contrast, in the 1000 mg/kg dose group, the parent compounds recovered in feces accounted for 48–61% of total intake. Biphenylhydrazine glucuronide accounted for 4.7–5.6% of total intake. The major components recovered in urine were conjugates of p,p-biphenyl or sulfates of p,p'-biphenyl and 4-hydroxybiphenyl. The total amount of these compounds accounted for 19-21% of the administered dose in the 10 mg/kg dose group and 6-7% of the administered dose in the 1000 mg/kg dose group. The major metabolites identified in bile were biphenyl, 4-hydroxyl (17-20% of AD in the 10 mg/kg group and 2.1-2.5% of AD in the 1000 mg/kg group), ... biphenyl, 4-hydroxyl, 4-methoxy (17-19% of AD in the 10 mg/kg group and 2.8% of AD in the 1000 mg/kg group) and ... biphenylhydrazine/glucuronide (9-12% of AD in the 10 mg/kg group and 9-13% of AD in the 1000 mg/kg group).

---

Biological half-life
Sprague-Dawley Crl:CD/BR rats (both male and female) were administered 10 or 1000 mg/kg of [(14)C]……biphenylhydrazine/……(radiochemical purity: >98%, specific activity: 34.4) mCi/mmol, labeled on the aromatic ring, /biphenylhydrazine/……(unlabeled……purity: 99.6%), by gavage. The following pharmacokinetic parameters were obtained: the tmax for men and women in the 10 mg/kg and 1000 mg/kg groups were 5 and 6 hours and 18-24 hours, respectively; the Cmax for men and women in the 10 mg/kg and 1000 mg/kg groups were 6.4 and 5.6 μg equivalents/g and 119 and 71 μg equivalents/g, respectively; and the t1/2 for men and women in the low-dose and high-dose groups were 11.5 and 13.3 hours and 12 and 15.6 hours, respectively.

Toxicity Summary
Identification and Uses: Biflorin is a solid. Used as an acaricide/mite-killer (insecticide). Human Exposure and Toxicity: No data available. Animal Studies: Biflorin has mild eye and skin irritation in rabbits. In dogs, at concentrations of 400 ppm and 1000 ppm, Biflorin caused myeloid hyperplasia and resulted in a dose-dependent decrease in erythrocyte count, and an increase in platelet count and serum bilirubin levels. In rats, the only observed developmental effects were mild prepuce-glans separation in male rats at concentrations of 80 ppm and 200 ppm, and vaginal perforation in female rats at a concentration of 200 ppm. This drug has not shown mutagenicity in Salmonella Typhimurium strains TA98, TA100, TA1535, and TA1537, or in Escherichia coli strain WP2 uvrA. No increase in the percentage of cells with chromosomal aberrations associated with treatment was observed in Chinese hamster ovary cells. Biflorin treatment did not lead to an increase in micronucleus numbers in mice. Ecotoxicity studies: Biflorin exhibits mild acute oral toxicity in poultry (LD50 = 1032 mg/kg) and moderate subacute ingestible toxicity in poultry (LC50 = 656-1862 ppm). Biflorin is virtually non-toxic to small mammals (LD50 > 5000 mg/kg). Existing data indicate that Biflorin is acutely highly toxic to freshwater fish (LC50 = 0.58-76 ppm) and aquatic invertebrates (LC50/EC50 = 0.50 ppm). Interactions…We tested the ability of known organophosphates and carbamates to inhibit Biflorin activation, thereby reducing its acaricidal activity. Esterase activity was measured in vivo after mites were pre-exposed to organophosphates and carbamates. The results showed that the degree of esterase inhibition by different compounds was closely related to their ability to antagonize the effects of Biflorin on mites. These findings suggest that organophosphates and carbamates interfere with the efficacy of Biflorin, most likely through the inhibition of carboxylesterases responsible for activating the prodrug. Due to the strong antagonistic effect, mixtures of Biflorin with carbamate or organophosphate pesticides should not be used in the field. Furthermore, repeated application of organophosphate pesticides and Biflorin also poses practical risks. This study again demonstrates that understanding the mechanism of action is crucial for the rational development of resistance management strategies.

---

Non-human toxicity values
Oral LD50 in rats >5000 mg/kg
Dermal LD50 in rats >5000 mg/kg

[1]. Dekeyser, M.A., McDonald, P.T., and Angle, G.W., Jr. The discovery of bifenazate, a novel carbazate acaricide Chimia 57(11), 702-704 (2003).

[2]. A novel action of highly specific acaricide; bifenazate as a synergist for a GABA-gated chloride channel of Tetranychus urticae [Acari: Tetranychidae]. Neurotoxicology. 2012 Jun;33(3):307-13.

Biphenylhydrazine is a carboxylic acid ester prepared by the condensation of 2-(4-methoxy[1,1'-biphenyl]-3-yl)hydrazide carboxylic acid with 2-propanol. It is an acaricide, functioning similarly to carbazole acid, and is derived from the hydride of biphenyl. Biphenylhydrazine has been reported to be present in Ganoderma lucidum, with relevant data available. Biphenylhydrazine is an acaricide used to control mites on ornamental plants grown in greenhouses, shade houses, nurseries, fields, gardens, and indoor landscaping. Biphenylhydrazine exhibits low acute toxicity (Class IV) through all routes of exposure and has no evidence of skin sensitization. It is non-irritating to the skin and has minimal eye irritation. In a series of necessary mutagenicity studies, biphenylhydrazine was negative for mutagenicity.

C17H20N2O3

300.3523

300.147

149877-41-8

176879

Light yellow to yellow solid powder

1.2±0.1 g/cm3

122℃

1.578

3.12

2

4

6

22

343

0

VHLKTXFWDRXILV-UHFFFAOYSA-N

InChI=1S/C17H20N2O3/c1-12(2)22-17(20)19-18-15-11-14(9-10-16(15)21-3)13-7-5-4-6-8-13/h4-12,18H,1-3H3,(H,19,20)

propan-2-yl N-(2-methoxy-5-phenylanilino)carbamate

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 : ≥ 250 mg/mL (~832.36 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (6.93 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 20.8 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.

---

Solubility in Formulation 2: ≥ 2.08 mg/mL (6.93 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

---

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