| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
| Targets |
The primary molecular targets of NSC 4810 have been identified as cytochrome P450 enzymes (for which it serves as a substrate in vivo) and acetylcholinesterase (for which it serves as a substrate in vitro). Cytochrome P450 enzymes are a family of heme-containing monooxygenases responsible for the oxidative metabolism of xenobiotics and endogenous compounds. As a substrate for P450 enzymes, NSC 4810 is metabolized to products that can be quantified to assess enzyme activity. Acetylcholinesterase is a serine hydrolase that terminates neurotransmission at cholinergic synapses by hydrolyzing acetylcholine. As a substrate for acetylcholinesterase, NSC 4810 is hydrolyzed by the enzyme, and the reaction products can be detected to measure enzyme activity. The compound's amide structure allows it to be recognized and processed by these enzymes, making it a useful probe for studying enzyme function, inhibition, and regulation in biological systems.
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| ln Vitro |
In vitro biological activity data for NSC 4810 are primarily related to its function as a substrate for acetylcholinesterase. In acetylcholinesterase assays, NSC 4810 is hydrolyzed by the enzyme, and the rate of hydrolysis can be measured to determine enzyme activity and to screen for potential inhibitors. The compound's hydrolysis by acetylcholinesterase can be monitored using various detection methods including spectrophotometric, fluorometric, or chromatographic techniques. The specific kinetic parameters (Km, Vmax) of NSC 4810 for acetylcholinesterase would need to be determined experimentally. As an amide substrate, it may be processed by other esterases and amidases as well, contributing to its overall in vitro profile. The compound may also exhibit other biological activities depending on the cell type and assay conditions, but its primary utility is as a research tool for enzyme studies rather than as a therapeutic agent.
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| ln Vivo |
In vivo activity of NSC 4810 relates to its function as a substrate for cytochrome P450 enzymes. When administered to animals, NSC 4810 is metabolized by hepatic cytochrome P450 enzymes, and the resulting metabolites can be detected and quantified in blood, urine, or tissues. This makes the compound useful for studying P450 enzyme activity in vivo, assessing drug-drug interactions, and evaluating the effects of various treatments or disease states on P450-mediated metabolism. The compound may also be used in pharmacokinetic studies to investigate the metabolic clearance and disposition of amide-containing compounds. Specific in vivo studies involving NSC 4810 would typically involve administration to rodents (mice or rats) via oral gavage, intraperitoneal injection, or intravenous injection, followed by collection of blood and tissue samples at various time points for metabolite analysis by HPLC or LC-MS/MS.
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| Enzyme Assay |
For in vitro enzyme assays with acetylcholinesterase substrates like NSC 4810, the following protocol is typically used: acetylcholinesterase (from Electrophorus electricus or human recombinant) is dissolved in 50 mM phosphate buffer (pH 7.4) at a concentration of 0.1-0.5 U/mL. The test compound is dissolved in DMSO and diluted in assay buffer to final concentrations ranging from 0.01 to 1000 μM. The reaction is initiated by adding the enzyme to the substrate solution in a 96-well plate, and the hydrolysis product is detected using a chromogenic or fluorogenic reagent. For Ellman's method, acetylthiocholine is used as a reference substrate, and the production of thiocholine is monitored by reaction with DTNB to produce a yellow color measured at 412 nm. For NSC 4810, hydrolysis of the amide bond releases a product that can be quantified by HPLC or LC-MS. Incubation is carried out at 25-37°C for 5-30 minutes, and the reaction is stopped by adding a stop solution. Kinetic parameters are calculated by fitting the data to Michaelis-Menten equation using nonlinear regression.
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| Cell Assay |
For in vitro cell-based assays with amide compounds like NSC 4810, the following typical protocol is used: human hepatocytes or liver microsomes are used to study cytochrome P450-mediated metabolism. Hepatocytes are cultured in William's E medium supplemented with 10% FBS and antibiotics at 37°C in 5% CO₂ for 24-48 hours to allow attachment and recovery. The test compound is added at concentrations of 1-100 μM, and cells are incubated for 0.5-4 hours. At various time points, aliquots of the medium are collected and analyzed by LC-MS/MS for the parent compound and metabolites. For cytotoxicity assessment, HepG2 or other cell lines are seeded in 96-well plates and treated with the compound at 0.1-200 μM for 24-72 hours, followed by MTT assay. For acetylcholinesterase activity in cell lysates, cells are lysed in RIPA buffer and the lysate is incubated with the substrate in 96-well plates, with the reaction product measured spectrophotometrically or fluorometrically.
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| Animal Protocol |
For in vivo animal studies with compounds like NSC 4810, the following general protocol is used: male or female Sprague-Dawley rats (6-8 weeks old, 180-220 g) or C57BL/6 mice (6-8 weeks old, 20-25 g) are used. The test compound is formulated in a suitable vehicle (e.g., 0.5% methylcellulose, PEG400, or saline) and administered via oral gavage at doses of 1-50 mg/kg. For intravenous administration, the compound is dissolved in saline or a mixture of saline and co-solvents and injected via the tail vein. Blood samples (100-200 μL) are collected from the retro-orbital plexus or tail vein at predetermined time points (0, 0.25, 0.5, 1, 2, 4, 6, 8, 12, and 24 hours post-dose). Plasma is separated by centrifugation and stored at -80°C until analysis. Urine and feces may also be collected for metabolism studies. Samples are analyzed by LC-MS/MS to determine the concentration of the parent compound and its metabolites. Pharmacokinetic parameters (Cmax, Tmax, AUC, t½, clearance, volume of distribution) are calculated using non-compartmental analysis. For P450 activity studies, the compound is administered and the rate of metabolite formation is determined.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of NSC 4810 have not been fully characterized in the literature. However, based on its physicochemical properties (molecular weight 239.74 g/mol, LogP ~3.2-3.8), the compound is expected to have moderate oral bioavailability and good membrane permeability. As an amide substrate for cytochrome P450 enzymes, it is likely metabolized by oxidative pathways including N-dealkylation and aromatic hydroxylation, mediated primarily by CYP3A4 and CYP2D6. The compound may also undergo hydrolysis by amidases to release the corresponding carboxylic acid and amine. Plasma protein binding is predicted to be moderate (70-85%) due to its lipophilic nature. The elimination half-life is estimated to be 2-4 hours in rats based on similar amide compounds. The compound is expected to be distributed to tissues with good perfusion including liver, kidney, and lung, and may have limited penetration into the central nervous system depending on its ability to cross the blood-brain barrier.
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| Toxicity/Toxicokinetics |
The toxicity profile of NSC 4810 has not been systematically evaluated. As an amide compound used as a research tool, its toxicological properties are not well-documented. However, benzamide and substituted benzamide derivatives generally require careful toxicity assessment. The chloro substituent may contribute to the compound's reactivity and potential toxicity. Some chlorinated benzamides have shown hepatotoxicity and nephrotoxicity in animal studies at high doses. The compound should be handled with appropriate safety precautions in a fume hood with personal protective equipment. For any therapeutic development, comprehensive toxicology studies including acute toxicity (LD50 determination in rodents), sub-chronic toxicity (28-day repeat-dose study), and genotoxicity testing (Ames test, micronucleus test) would be required. The compound's status as a P450 substrate suggests potential for drug-drug interactions when co-administered with other P450 substrates or inhibitors.
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| Additional Infomation |
NSC 4810 (CAS# 79606-45-4) is an amide bioactive compound with the molecular formula C13H18ClNO. It is also known as 4-chloro-N,N-diisopropylbenzamide, OR59402, and SKL760. It has been used as a substrate for cytochrome P450 enzymes in vivo and as a substrate for acetylcholinesterase in vitro, making it a valuable tool compound for studying enzyme kinetics, drug metabolism, and neurotransmitter regulation. The compound has a purity of ≥98% and an appearance of off-white solid with a melting point of 87-89°C. Future research could explore its utility in high-throughput screening for enzyme inhibitors, its metabolism by various P450 isoforms, and its potential as a lead compound for developing novel therapeutics targeting acetylcholinesterase or other enzymes.
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| Molecular Formula |
C13H18NOCL
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| Molecular Weight |
239.74112
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| Exact Mass |
239.108
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| CAS # |
79606-45-4
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| PubChem CID |
241026
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| Appearance |
Typically exists as solid at room temperature
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| Density |
1.072g/cm3
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| Boiling Point |
347.7ºC at 760 mmHg
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| Flash Point |
164.1ºC
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| Index of Refraction |
1.52
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| LogP |
3.599
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
1
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
16
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| Complexity |
224
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC(C)N(C(C)C)C(=O)C1=CC=C(C=C1)Cl
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| InChi Key |
MGPPJDHMGGQMSG-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C13H18ClNO/c1-9(2)15(10(3)4)13(16)11-5-7-12(14)8-6-11/h5-10H,1-4H3
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| Chemical Name |
4-chloro-N,N-di(propan-2-yl)benzamide
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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
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| Solubility (In Vivo) |
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.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] Oral Formulations
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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.1712 mL | 20.8559 mL | 41.7119 mL | |
| 5 mM | 0.8342 mL | 4.1712 mL | 8.3424 mL | |
| 10 mM | 0.4171 mL | 2.0856 mL | 4.1712 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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