1. Metabolic Enzyme/Protease
  2. Endogenous Metabolite
  3. AFMK

AFMK, antioxidant metabolite of Melatonin, attenuates X-ray-induced oxidative damage to DNA, proteins and lipids in mice. AFMK is a poorer scavenger. The pKa of AFMK at physiological pH is 8.7. Antioxidant capacity. AFMK improves the anti-tumor effect of Gemcitabine in PANC-1 cells through the modulation of apoptotic pathway.

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AFMK Chemical Structure

AFMK Chemical Structure

CAS No. : 52450-38-1

Size Price Stock Quantity
Solid + Solvent
10 mM * 1 mL in DMSO
ready for reconstitution
USD 264 In-stock
Solution
10 mM * 1 mL in DMSO USD 264 In-stock
Solid
1 mg USD 80 In-stock
5 mg USD 240 In-stock
10 mg USD 390 In-stock
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Based on 1 publication(s) in Google Scholar

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  • Biological Activity

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Description

AFMK, antioxidant metabolite of Melatonin, attenuates X-ray-induced oxidative damage to DNA, proteins and lipids in mice. AFMK is a poorer scavenger. The pKa of AFMK at physiological pH is 8.7. Antioxidant capacity[1][2]. AFMK improves the anti-tumor effect of Gemcitabine in PANC-1 cells through the modulation of apoptotic pathway[3].

IC50 & Target

Human Endogenous Metabolite

 

In Vitro

AFMK is one of the metabolites of melatonin and can be formed by both enzymatic or pseudoenzymatic and nonenzymatic metabolic pathways[1].
AFMK pretreatment significantly inhibits DNA damage. AFMK shows a very high level of in vitro hydroxyl radical scavenging potential which was measured by an electron spin resonance (ESR) study. IC50 values resulting from ESR analysis was 338.08 nM. AFMK, a melatonin metabolite, is a sparingly investigated biogenic amine[2].
AFMK administered to PANC-1 in combination with Gemcitabine inhibits the production of HSP70 and cIAP-2 as compared to the results obtained with Gemcitabine alone[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Western Blot Analysis[3]

Cell Line: Human pancreatic carcinoma cell line (PANC-1)
Concentration: 0.001, 0.1, 10, 1000 nM
Incubation Time:
Result: 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).
In Vivo

AFMK is a potent antioxidant in vivo. AFMK significantly reverses radiation-induced decline in the total antioxidant capacity of plasma in mice[2].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model: Male C57BL mice 8 wk of age[2]
Dosage: 10 mg/kg body weight
Administration: Intraperitoneal injection
Result: Radiation-induced decline in the total antioxidant capacity of plasma was significantly reversed in AFMK pretreated mice.
AFMK-pretreated irradiated groups showed a significantly lower value of comet tail length and % DNA in tail.
Molecular Weight

264.28

Formula

C13H16N2O4

CAS No.
Appearance

Solid

Color

Light yellow to light brown

SMILES

CC(NCCC(C1=CC(OC)=CC=C1NC=O)=O)=O

Structure Classification
Initial Source
Shipping

Room temperature in continental US; may vary elsewhere.

Storage
Powder -20°C 3 years
In solvent -80°C 6 months
-20°C 1 month
Solvent & Solubility
In Vitro: 

DMSO : 50 mg/mL (189.19 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 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
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

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In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.5 mg/mL (9.46 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 2.5 mg/mL (9.46 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

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(per animal)

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Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation

Purity: 99.77%

References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 3.7839 mL 18.9193 mL 37.8387 mL 94.5966 mL
5 mM 0.7568 mL 3.7839 mL 7.5677 mL 18.9193 mL
10 mM 0.3784 mL 1.8919 mL 3.7839 mL 9.4597 mL
15 mM 0.2523 mL 1.2613 mL 2.5226 mL 6.3064 mL
20 mM 0.1892 mL 0.9460 mL 1.8919 mL 4.7298 mL
25 mM 0.1514 mL 0.7568 mL 1.5135 mL 3.7839 mL
30 mM 0.1261 mL 0.6306 mL 1.2613 mL 3.1532 mL
40 mM 0.0946 mL 0.4730 mL 0.9460 mL 2.3649 mL
50 mM 0.0757 mL 0.3784 mL 0.7568 mL 1.8919 mL
60 mM 0.0631 mL 0.3153 mL 0.6306 mL 1.5766 mL
80 mM 0.0473 mL 0.2365 mL 0.4730 mL 1.1825 mL
100 mM 0.0378 mL 0.1892 mL 0.3784 mL 0.9460 mL
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AFMK Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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AFMK
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HY-113314
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