Gibberellin biosynthesis[1]

The activity of JMJD2A demethylase is inhibited by prohexadione (1 mM)[2]. Mouse neurospheres are induced to differentiate into neurons by prohexadione (1-2 mM)[2]. In soybean seedlings, prohexadione (100 mg/L) sprayed on the leaves reduces saline-alkali stress[3]. In potted apple trees, prohexadione increases structural resistance against fire blight infection[4].

Absorption, Distribution and Excretion
In rat metabolism studies, cyclohexanone calcium was rapidly absorbed, reaching peak tissue/carcass concentrations within 30 minutes; however, absorption saturated at the highest dose. The test substance did not accumulate in tissues. For low-dose animals, renal excretion was the primary clearance route. At high doses, fecal excretion became the primary clearance route. The main excretory metabolites (urine and feces) were identified as free acids. Metabolism/Metabolites Male and female Fisher 344 rats were administered 50 or 500 mg/kg of [14C]-BX-112 (radiolabeled at positions 3 or 5 of the cyclohexene ring) (batch number CP-1107, radiochemical purity: 98.9%, specific activity: 97.8 mCi/mg) by gavage. In groups B (50 mg/kg) and D (500 mg/kg) (mass balance study), urine, feces, and cage cleaning fluid samples were collected from 5 animals of each sex in each group for up to 7 days. In group C1, 5 animals of each sex were pretreated with unlabeled BX-112 technology (purity: 93.8%) for 14 days, followed by administration of 50 mg/kg of radiolabeled material. Urine, feces, and cage cleaning fluid samples were collected within 7 days after the last administration. Samples collected from 0 to 48 hours after administration were analyzed to identify specific metabolites. In addition, radiolabeled material was extracted from liver and kidney tissues of male animals treated with 50 mg/kg (group B3) and 500 mg/kg (group D3), respectively, and the animals were sacrificed 0.5 hours after administration. The test substance was primarily excreted in urine and feces as a free acid (50 mg/kg: 38.3%–39.1%, 500 mg/kg: 60.9%–68.0%, 50 mg/kg repeated administration: 44.2%–53.7%). Another compound isolated from urine was preliminarily identified as an ester glucuronide of BX-112. The total percentages of the test substance's compound were 20.4% and 21.7% of the administered dose in males and females (50 mg/kg group), 7.3% and 3.7% (50 mg/kg repeated administration group), and 23.2% and 17.7% (50 mg/kg repeated administration group), respectively. The only other metabolite identified was recovered from the organic phase of the fecal extract. This substance was depropionyl free acid of BX-112. Its content did not exceed 2.3% of the administered dose in all test groups. In both liver and kidney, the predominant recovered substance at both dose levels was the free acid of BX-112 (liver: 81.2% and 93.1% of the recovered radiolabeled substance in the 50 mg/kg and 500 mg/kg groups, respectively; kidney: 70.1% and 91.8% of the recovered labeled substance in the 50 mg/kg and 500 mg/kg groups, respectively). The appendix notes that two previously unknown compounds were methyl esters of the test substance and its depropionyl metabolite, considered experimental errors. The appendix also provides further information on the separation of radiolabeled compounds from feces; the separated compounds were predominantly free acids of the parent compound, consistent with the initial report. Furthermore, the use of a C18 HPLC column instead of a C8 column resulted in a different ratio of parent compound to conjugate compared to the ratio observed in the original analysis; further analysis of this "conjugate" by FAB-MS and atmospheric piezo-spray HPLC/MS did not definitively identify the compound.

Toxicity Data
LC50 (Rat) > 4,210 mg/m³/4h

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5000 mg/kg LD50 (Rat, Dermal Contact) > 2000 mg/kg LD50 (Mouse, Oral) > 5000 mg/kg LC50 (Rat, Inhalation) > 4.21 mg/L Air/4 hours (Systemic)

[1]. Effect of a plant growth regulator prohexadione-calcium on insect pests of apple and pear. J Econ Entomol. 2005 Apr;98(2):423-31.

[2]. Prohexadione, a plant growth regulator, inhibits histone lysine demethylases and modulates epigenetics. Toxicol Rep. 2014 Nov 4;1:1152-1161.

[3]. Prohexadione-calcium alleviates saline-alkali stress in soybean seedlings by improving the photosynthesis and up-regulating antioxidant defense. Ecotoxicol Environ Saf. 2021 Sep 1;220:112369.

[4]. Evidence that prohexadione-calcium induces structural resistance to fire blight infection. Phytopathology. 2009 May;99(5):591-6.

Prohexadione is a 4-oxomonocarboxylic acid formed by substituting carbonyl groups at the 3 and 5 positions and a propionyl group at the 4 position of cyclohexane carboxylic acid. It is a plant growth regulator, commonly used as an anti-lodging agent for small grains in the form of its corresponding calcium salt (called calcium propylcyclohexanone). It has various uses, including as an agrochemical, plant growth regulator, and gibberellin biosynthesis inhibitor. It is a 4-oxomonocarboxylic acid and a β-triketone, and is also the conjugate acid of propylcyclohexanone (2-).

C10H12O5

212.20

212.068

88805-35-0

Prohexadione calcium;127277-53-6

184900

Fine white powder
Pale yellow brown colored fine powder

1.33 g/cm3

454.9ºC at 760 mmHg

>360 °C
Yellow powder with sweet smell. Melting point >360 °C. Hardly soluble in organic solvents. /Technical/

243.1ºC

1.512

0.214

1

5

3

15

313

0

O=C1C(C(=O)CC)C(=O)CC(C(O)=O)C1

BUCOQPHDYUOJSI-UHFFFAOYSA-N

InChI=1S/C10H12O5/c1-2-6(11)9-7(12)3-5(10(14)15)4-8(9)13/h5,9H,2-4H2,1H3,(H,14,15)

3,5-dioxo-4-propanoylcyclohexane-1-carboxylic acid

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.)