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AFMK

Cat No.:V72320 Purity: ≥98%
AFMK is an antioxidant metabolite of melatonin that attenuates X-ray-induced oxidative damage to DNA, protein, and lipids in mice.
AFMK
AFMK Chemical Structure CAS No.: 52450-38-1
Product category: Endogenous Metabolite
This product is for research use only, not for human use. We do not sell to patients.
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Product Description
AFMK is an antioxidant metabolite of melatonin that attenuates X-ray-induced oxidative damage to DNA, protein, and lipids in mice. AFMK is a less effective detergent with a pKa of 8.7 at physiological pH. Has anti-oxidant effect. AFMK can enhance the anti-tumor effect of Gemcitabine in PANC-1 cells by regulating the apoptosis pathway.
Biological Activity I Assay Protocols (From Reference)
Targets
Human Endogenous Metabolite
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].
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].
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).
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.
References

[1]. On the free radical scavenging activities of melatonin's metabolites, AFMK and AMK. J Pineal Res. 2013 Apr;54(3):245-57.

[2]. AFMK, a melatonin metabolite, attenuates X-ray-induced oxidative damage to DNA, proteins and lipids in mice. J Pineal Res. 2007 Apr;42(4):386-93.

[3]. Melatonin and its metabolite N1-acetyl-N2-formyl-5-methoxykynuramine (afmk) enhance chemosensitivity to gemcitabine in pancreatic carcinoma cells (PANC-1). Pharmacol Rep. 2018 Dec;70(6):1079-1088.

Additional Infomation
N-gamma-Acetyl-N-2-Formyl-5-Methoxykynurenamine is an aromatic ketone.
These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C13H16N2O4
Molecular Weight
264.28
Exact Mass
264.11
CAS #
52450-38-1
PubChem CID
171161
Appearance
Light yellow to light brown solid powder
Density
1.2±0.1 g/cm3
Boiling Point
589.4±50.0 °C at 760 mmHg
Melting Point
138-140ºC
Flash Point
310.3±30.1 °C
Vapour Pressure
0.0±1.7 mmHg at 25°C
Index of Refraction
1.560
LogP
0.82
Hydrogen Bond Donor Count
2
Hydrogen Bond Acceptor Count
4
Rotatable Bond Count
6
Heavy Atom Count
19
Complexity
333
Defined Atom Stereocenter Count
0
InChi Key
JYWNYMJKURVPFH-UHFFFAOYSA-N
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)
Chemical Name
N-[3-(2-formamido-5-methoxyphenyl)-3-oxopropyl]acetamide
HS Tariff Code
2934.99.9001
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)
Solubility Data
Solubility (In Vitro)
DMSO: 50 mg/mL (189.19 mM)
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.

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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.
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 corn oil and mix evenly.


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

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What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
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What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
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Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
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In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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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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