3-Methyladenine (3-MA)

Biological Activity

3-Methyladenine is a selective autophagy and phosphoinositide 3-kinase (PI3K) inhibitor for Vps34 and PI3Kγ with IC50 of 25 and 60 nM, respectively. 3-Methyladenine significantly shortens the duration of nocodazole-induced-prometaphase arrest. 3-Methyladenine inhibits class I and class III PI3K in different temporal patterns. 3-Methyladenine blocks autophagy through its effect on class III phosphatidylinositol 3-kinase (PI3K), the activity of which is required for the nucleation and assembly of the initial phagophore membrane. 3-Methyladenine treatment does not alter the degree of hemorrhage compared with the SAH group.

Product Citations

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  Oxid Med Cell Longev. 2021 Jun 14;2021:6662225.

  Impaired Autophagy Induced by oxLDL/ β 2GPI/anti- β 2GPI Complex through PI3K/AKT/mTOR and eNOS Signaling Pathways Contributes to Endothelial Cell Dysfunction
  3-Methyladenine (3-MA) purchased from AbMole

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  BMC Cancer. 2021 Feb 25;21(1):195.

  Blockage of AMPK-ULK1 pathway mediated autophagy promotes cell apoptosis to increase doxorubicin sensitivity in breast cancer (BC) cells: an in vitro study
  3-Methyladenine (3-MA) purchased from AbMole

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  Int J Neurosci. 2020 Mar 11;1-15.

  Autophagy-deficiency in Bone Marrow Mononuclear Cells From Patients With Myasthenia Gravis: A Possible Mechanism of Pathogenesis.
  3-Methyladenine (3-MA) purchased from AbMole

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  Front Pharmacol 2018 May 14;14;9:334.

  Resveratrol Sensitizes Carfilzomib-Induced Apoptosis via Promoting Oxidative Stress in Multiple Myeloma Cells
  3-Methyladenine (3-MA) purchased from AbMole

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  Mol Immunol. 2017 Jul;87:132-140.

  IL-37 induces autophagy in hepatocellular carcinoma cells by inhibiting the PI3K/AKT/mTOR pathway
  3-Methyladenine (3-MA) purchased from AbMole

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  Int J Clin Exp Med. 2017;10(5):7553-7562.

  Combination of icotinib and wogonin induces apoptosis and autophagy to overcome acquired resistance in lung cancer harbouring EGFR T790M mutation
  3-Methyladenine (3-MA) purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Front Pharmacol (2018). Figure 6. 3-methyladenine (AbMole BioScience, Houston, TX, USA)
	Method	Intracellular ROS Production
	Cell Lines	MM cells
	Concentrations	1 mmol/L
	Incubation Time	2 h
	Results	However, RSV/CFZ combination increased levels of LC3-II and p62/SQSTM1. Higher levels of LC3-II could be partially blocked by the autophagy inhibitor 3-MA, but not for p62/SQSTM1

	Source	Int J Clin Exp Med (2017) . Figure 6. 3-methyladenine (3-MA) (Abmole Bioscience Inc. Houston, TX, USA)
	Method	Evaluation of the inhibitory activity
	Cell Lines	Human NSCLC cell line NCI-H1975
	Concentrations	100 mM
	Incubation Time
	Results	In addition, the suppression of mTOR phosphorylation decreased the percentage of early apoptotic cells (Figure 6A and 6B) and caspase 3 cleavage (Figure 6C).

	Source	Int J Clin Exp Med (2017) . Figure 5. 3-methyladenine (3-MA) (Abmole Bioscience Inc. Houston, TX, USA)
	Method	Evaluation of the inhibitory activity
	Cell Lines	Human NSCLC cell line NCI-H1975
	Concentrations	100 mM
	Incubation Time
	Results	As a control, 3-MA was used to prevent mTOR phosphorylation. The suppression of mTOR phosphorylation decreased the number of intracellular autophagosomes increased by treatment with wogonin plus icotinib combination (Figure 5B).

	Source	Mol Immunol (2017) . Figure 2. 3-methyladenine (Abmole Bioscience, USA)
	Method	IL-37 induces autophagy in SMMC-7721 and Huh-7 cells.
	Cell Lines	SMMC-7721 and Huh-7 cells
	Concentrations	5 mmol/l
	Incubation Time
	Results	The LC3-II/LC3-I ratio was significantly decreased in IL-37/3-MA-co-treated SMMC-7721 and Huh-7 cells and increased in IL-37/CQ-co-treated SMMC-7721 and Huh-7 cell (Fig. 2E and F). Together, these data further indicate that IL-37 induces autophagy in SMMC-7721 and Huh-7 cells.

Protocol

Cell Experiment
Cell lines	U251 cells
Preparation method	Western blotting was performed to determine the expression levels of ubiquitinated proteins, PDI and Grp78. The expression levels of ubiquitinated PDI and Grp78 were increased following the inhibition of autophagy by treatment with 3-MA
Concentrations	10 mM
Incubation time	12 h

Animal Experiment
Animal models	Male Wistar rats
Formulation	saline
Dosages	3.5 mg/kg
Administration	i.v.

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

Chemical Information

References

[1] Zhang R, et al. Mol Med Rep. Inhibition of autophagy using 3-methyladenine increases cisplatin-induced apoptosis by increasing endoplasmic reticulum stress in U251 human glioma cells.

[2] Zhu B, et al. Int J Clin Exp Med. 3-methyladenine, an autophagic inhibitor, attenuates therapeutic effects of sirolimus on scopolamine-induced cognitive dysfunction in a rat model.

[3] Hualei Guo, et al. Cardiovasc Drυgs Ther. Resveratrol protects HUVECs from oxidized-LDL induced oxidative damage by autophagy upregulation via the AMPK/SIRT1 pathway

[4] Ying Xie, et al. Mol Med Rep. Protective role of autophagy in AGE-induced early injury of human vascular endothelial cells