| Size | Price | Stock | Qty |
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| Targets |
Human Endogenous Metabolite
AFMK targets oxidative stress pathways as an antioxidant metabolite of melatonin. It functions as a free radical scavenger, protecting cells from oxidative and nitrosative stress. AFMK may also be measured in plasma as an index of melatonin synthesis and metabolism. |
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| ln Vitro |
One of the metabolites of melatonin, AFMK is produced by a variety of metabolic routes, including nonenzymatic, pseudoenzymatic, and enzymatic ones[1]. Pretreatment with AFMK dramatically reduces DNA damage. The electron spin resonance (ESR) investigation used to test AFMK's in vitro hydroxyl radical scavenging potential revealed an extremely high level of activity. The ESR study yielded IC50 values of 338.08 nM. AFMK, a metabolite of melatonin, is a biogenic amine that is rarely studied[2]. In comparison to the outcomes achieved with Gemcitabine alone, AFMK given to PANC-1 in combination with Gemcitabine suppresses the synthesis of HSP70 and cIAP-2[3].
In vitro, AFMK has antioxidant and free radical scavenging activities in several experimental models. It is a potent antioxidant that attenuates oxidative damage to DNA, proteins, and lipids. AFMK is a poorer scavenger compared to some other antioxidants, with a pKa of 8.7 at physiological pH. It protects cells from oxidative and nitrosative stress. |
| ln Vivo |
In vivo, AFMK is a strong antioxidant. AFMK dramatically reverses the decrease in mice's plasma's overall antioxidant capacity that is brought on by radiation[2].
In vivo, AFMK attenuates X-ray-induced oxidative damage to DNA, proteins and lipids in mice. It is a major metabolite of melatonin produced via enzymatic and oxidative pathways. AFMK may be measured in plasma as an index of melatonin synthesis and metabolism. The compound has antioxidant activity in vivo. |
| Enzyme Assay |
In vitro antioxidant assays for AFMK involve incubating the compound with free radical generators such as DPPH, ABTS, or peroxyl radicals. Radical scavenging activity is measured spectrophotometrically by monitoring the decrease in absorbance. For cell-based assays, cells are treated with AFMK and oxidative stress is induced by H₂O₂ or X-ray irradiation. Protection against oxidative damage is assessed by measuring ROS levels, DNA damage, and lipid peroxidation.
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| Cell Assay |
Western Blot Analysis[3]
Cell Types: Human pancreatic carcinoma cell line (PANC-1) Tested Concentrations: 0.001, 0.1, 10, 1000 nM Incubation Duration: Experimental Results: Augmented the inhibitory effects on HSP70 expression from 0.47 (Gemcitabine alone) to 0.13 (10 nM AFMK), 0.08 (0.1 nM AFMK) and 0.01 (0.001 nM AFMK). For in vitro cell assays, cells are cultured in medium supplemented with AFMK at concentrations of 1-100 µM. Cell viability is assessed by MTT assay. ROS levels are measured using fluorescent probes such as DCFH-DA. Protection against oxidative damage is evaluated by measuring DNA damage (comet assay), protein carbonylation, and lipid peroxidation (MDA). The compound's antioxidant effects are assessed in cells exposed to oxidants or radiation. |
| Animal Protocol |
Animal/Disease Models: Male C57BL mice 8 wk of age[2]
Doses: 10 mg/kg body weight Route of Administration: intraperitoneal (ip) injection Experimental Results: Radiation-induced decline in the total antioxidant capacity of plasma was Dramatically reversed in AFMK pretreated mice. AFMK-pretreated irradiated groups demonstrated a Dramatically lower value of comet tail length and % DNA in tail. For in vivo animal studies, AFMK is administered to mice via oral gavage or intraperitoneal injection at doses of 10-100 mg/kg. The compound's protective effects against X-ray-induced oxidative damage are evaluated by measuring DNA, protein, and lipid damage in tissues. Plasma AFMK levels are quantified by LC-MS/MS as an index of melatonin metabolism. Tissue distribution and antioxidant effects are assessed. |
| ADME/Pharmacokinetics |
AFMK (MW 264.28) has a molecular formula of C13H16N2O4. It is soluble in DMSO and organic solvents. The compound is stable under standard storage conditions. As a melatonin metabolite, it is produced through enzymatic and oxidative pathways. The pKa of AFMK at physiological pH is 8.7. It is an endogenous biogenic amine.
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| Toxicity/Toxicokinetics |
The compound is for research use only. No specific toxicity data are available. As an endogenous metabolite of melatonin, it is expected to have low toxicity at physiological concentrations. High doses may affect melatonin metabolism and oxidative stress pathways. Standard safety precautions should be followed.
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| References |
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| Additional Infomation |
N-γ-acetyl-N-2-formyl-5-methoxykynurenamine is an aromatic ketone.
AFMK (CAS# 52450-38-1) is an endogenous metabolite of melatonin and a research reagent used primarily in studies of oxidative stress, antioxidant defense, and melatonin metabolism. It has not been investigated in clinical trials nor approved as a therapeutic drug. The compound is used as an index of melatonin synthesis and metabolism in plasma and in studies of radiation-induced oxidative damage. It is a valuable tool for studying the antioxidant properties of melatonin metabolites. |
| Molecular Formula |
C13H16N2O4
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|---|---|
| Molecular Weight |
264.28
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| Exact Mass |
264.11
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| CAS # |
52450-38-1
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| PubChem CID |
171161
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| Appearance |
Light yellow to light brown solid powder
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| Density |
1.2±0.1 g/cm3
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| Boiling Point |
589.4±50.0 °C at 760 mmHg
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| Melting Point |
138-140ºC
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| Flash Point |
310.3±30.1 °C
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| Vapour Pressure |
0.0±1.7 mmHg at 25°C
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| Index of Refraction |
1.560
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| LogP |
0.82
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
19
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| Complexity |
333
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC(=O)NCCC(=O)C1=C(C=CC(=C1)OC)NC=O
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| InChi Key |
JYWNYMJKURVPFH-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C13H16N2O4/c1-9(17)14-6-5-13(18)11-7-10(19-2)3-4-12(11)15-8-16/h3-4,7-8H,5-6H2,1-2H3,(H,14,17)(H,15,16)
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| Chemical Name |
N-[3-(2-formamido-5-methoxyphenyl)-3-oxopropyl]acetamide
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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) |
DMSO: 50 mg/mL (189.19 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (9.46 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 25.0 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: ≥ 2.5 mg/mL (9.46 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (9.46 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.7839 mL | 18.9193 mL | 37.8387 mL | |
| 5 mM | 0.7568 mL | 3.7839 mL | 7.5677 mL | |
| 10 mM | 0.3784 mL | 1.8919 mL | 3.7839 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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