Fungal targets involved in spore germination and infection structure development. In mammalian cells, it induces phosphorylation of JNK, activates the p38 pathway, decreases Bcl-2, activates caspase-3, decreases complex I activity, and increases autophagy and apoptosis.

In vitro, Fluazinam exhibits broad-spectrum antifungal activity. It has a strong inhibition effect on mycelial growth of S. sclerotiorum populations. It induces phosphorylation of JNK, activates the p38 pathway, decreases Bcl-2, activates caspase-3, decreases complex I activity, and increases autophagy and apoptosis in mammalian cells.

In vivo, Fluazinam is used as a protective fungicide to control fungal diseases in crops. It is orally active. Detailed dose-response and pharmacokinetic data in animals are limited as it is primarily an agricultural fungicide.

No specific cell-free enzyme assay protocol is detailed for Fluazinam. For fungicides, typical cell-free assays include fungal enzyme inhibition studies (e.g., mitochondrial complex I activity assays) or spore germination inhibition assays.

Cellular antifungal activity is assessed in fungal cultures. Fluazinam is tested against various fungal pathogens using agar dilution or broth microdilution methods to determine MIC values. Mycelial growth inhibition is measured. Apoptosis and autophagy induction can be assessed in mammalian cell lines.

In vivo efficacy is evaluated in agricultural settings or in plant pathology studies. Fluazinam is applied as a foliar spray or soil treatment. Endpoints include disease severity scores, fungal lesion counts, and crop yield.

Absorption, Distribution and Excretion
Metabolites AMPA (4-chloro-N2-[3-chloro-5-(trifluoromethyl)-2-pyridyl]-3-nitro-5-(trifluoromethyl)-1,2-phenylenediamine), DAPA (3-chloro-2-(2,6-diamino-3-chloro-α,α,α-trifluoromethyl)pyridine), and some related conjugates and hydrolysis products were isolated, identified, and characterized from the urine, feces, and bile of radiolabeled fluazinam-treated rats. Fluazinam is almost entirely metabolized via hydroxylation followed by conjugation. No sex differences were observed. In rat metabolic studies, only 33–40% of the radiolabeled fluazinam was absorbed. The majority of the administered dose (>89%) was recovered in feces. The radioactive material detected in feces was primarily the unabsorbed parent compound. Very little was excreted in urine (<4%). However, the total radioactive material in bile accounted for 25-34% of the administered dose, indicating significant enterohepatic circulation. Chromatographic analysis revealed the presence of multiple metabolites in the bile.

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Metabolism/Metabolites
…Fluoridamine was almost entirely metabolized in the test rats via hydroxylation, followed by a conjugation reaction. No sex differences were observed.
Molecular weight: 465.1 g/mol; molecular formula: C13H4Cl2F6N4O4. CAS No.: 79622-59-6. IUPAC name: 3-chloro-N-[3-chloro-2,6-dinitro-4-(trifluoromethyl)phenyl]-5-(trifluoromethyl)pyridin-2-amine. Appearance: typically a solid. Storage: typical for agrochemicals.

Toxicity Data
LC50 (Rat) = 463 mg/m³

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Non-human Toxicity Values LD50 (Rats, Oral) >5000 mg/kg
LD50 (Rabbit, Dermal) >2000 mg/kg
Fluazinam has been evaluated by the WHO/JMPR for pesticide residues. It has established acceptable daily intake (ADI) and acute reference dose (ARfD) values. As an agricultural fungicide, it has moderate toxicity and appropriate safety precautions are recommended during handling.

[1]. Impact of fluazinam on morphological and physiological characteristics of Sclerotinia sclerotiorum. Pestic Biochem Physiol. 2019 Mar;155:81-89.

Fluazinam is an aminopyridine compound, chemically named 2-amino-5-(trifluoromethyl)pyridine, in which one amino hydrogen atom is replaced by 3-chloro-2,6-dinitro-4-(trifluoromethyl)phenyl. It is a fungicide used to control gray mold, downy mildew, and other fungal pathogens. Fluazinam possesses properties such as inducing apoptosis, sensitization, xenobiotic activity, environmental pollution, and antifungal effects. It is a C-nitro compound, a chloropyridine compound, an aminopyridine compound, a secondary amino compound, a monochlorobenzene compound, and a (trifluoromethyl)benzene compound. Fluazinam is a broad-spectrum fungicide widely used in agriculture. It belongs to the diarylamine class of compounds, more specifically, an arylaminopyridine compound. Its mechanism of action lies in its strong uncoupling ability against oxidative phosphorylation in mitochondria and its high reactivity with thiols. Unlike other uncoupling agents, it exhibits broad-spectrum activity against fungi and, due to its rapid metabolism into compounds without uncoupling activity, has extremely low toxicity to mammals. Fluazinam is a protective fungicide, but it is neither systemic nor curative. Its mechanism of action is to inhibit spore germination and the formation of infection structures. Although effective against a variety of fungi, it is less effective against rust and powdery mildew, and therefore has not been commercially used in cereal crops. Due to its activity against the zoospores of pathogens, fluazinam is widely used to control potato late blight (Phytophthora infestans), and is particularly effective in controlling infection of potato tubers.

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Mechanism of Action
Fluazinam is a lipophilic weak acid with strong uncoupling activity against mitochondria in vitro.

Fluazinam is an approved agricultural fungicide used in many countries. It is not a pharmaceutical drug and is not approved for human therapeutic use. It is used in integrated pest management strategies to prevent crop losses caused by fungal infections. Its mechanism involves inhibition of spore germination and infection structure development.

C13H4CL2F6N4O4

465.09

463.951

79622-59-6

91731

Light yellow to yellow solid powder

1.8±0.1 g/cm3

376.1±42.0 °C at 760 mmHg

113ºC

181.3±27.9 °C

0.0±0.9 mmHg at 25°C

1.571

8.19

1

12

2

29

628

0

UZCGKGPEKUCDTF-UHFFFAOYSA-N

InChI=1S/C13H4Cl2F6N4O4/c14-6-1-4(12(16,17)18)3-22-11(6)23-9-7(24(26)27)2-5(13(19,20)21)8(15)10(9)25(28)29/h1-3H,(H,22,23)

3-chloro-N-[3-chloro-2,6-dinitro-4-(trifluoromethyl)phenyl]-5-(trifluoromethyl)pyridin-2-amine

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 : ~125 mg/mL (~268.77 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.47 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.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (4.47 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.

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