Home PI3K/Akt/mTOR PI3K inhibitor - Autophagy inhibitor 3-MA (3-Methyladenine) For research use only.

3-MA (3-Methyladenine)

Synonyms: NSC 66389

3-MA (3-Methyladenine) is a selective PI3K inhibitor for Vps34 and PI3Kγ with IC50 of 25 μM and 60 μM in HeLa cells; blocks class I PI3K consistently, whereas suppression of class III PI3K is transient, and also blocks autophagosome formation. Solutions are unstable and should be fresh-prepared.

3-MA (3-Methyladenine) Chemical Structure

3-MA (3-Methyladenine) Chemical Structure

CAS No. 5142-23-4

Purity & Quality Control

Products often used together with 3-MA (3-Methyladenine)

Rapamycin

3-Methyladenine remarkably inhibits autophagy in colon tissues of LPS-treated mice, while Rapamycin induces autophagy.

Zhou M, et al. EBioMedicine. 2018 Sep;35:345-360.

Necrostatin-1

3-Methyladenine and Necrostatin-1 inhibit cell death of bone marrow macrophages (BMDM) induced by LPS/zVAD and PolyI: C/zVAD.

Chen YS, et al. Mol Cells. 2022 Apr 30;45(4):257-272.

Z-VAD-FMK

3-Methyladenine and Z-VAD-FMK combination confirm vital role of programmed cell death in pristimerin-mediated anti-cancer actions.

Al-Tamimi M, et al. Biomed Pharmacother. 2022 Dec;156:113950.

Fer-1 (Ferrostatin-1)

3-methyladenine and Ferrostatin-1 use abolishes acrylamide (ACR)-induced cell death in chondrocytes.

Wang H, et al. Exp Ther Med. 2023 Apr 12;25(6):246.

3-MA (3-Methyladenine) Related Products

Signaling Pathway

PI3K Signaling Pathways

PI3K Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
K562 Function Assay 10mM 1h decreases the expression of LC3-II and the formation of autophagosomes 21864037
Jurkat Function Assay 10mM 1h decreases the expression of LC3-II and the formation of autophagosomes 21864037
HeLa  Cytotoxicity Assay 2mM 24h inhibites the cytotoxicity of silibinin to HeLa cells. 21875385
PC12/TetOn Function Assay 0.1/1mM 18h leads to α-syn(WT) accumulation, toxicity, and oligomer formation  21906659
RMPI8226  Function Assay 5mM 1h suppresses the level of autophagy under nutrient depletion 21915620
MCF-7 Function Assay 10mM 48h blocks autophagy induced by bortezomib 21931937
HBx  Apoptosis Assay 10mM 48h DMSO increases cell death 22020078
Marc-145 Function Assay 5mM 12/24/36h reduces the PRRSV titers and the protein expression 22119900
U937 Function Assay 2mM 12h decreases the autophagy ratio  22155150
BGC-823 Function Assay 5mM 2h inhibits the rate of autophagic cells 22322152
A549 Function Assay 0.1mM 24h suppresses SU11274-induced cell death 22466960
pDCs Function Assay 10mM 0.5h  reduces the induction of IFN-α by ssRNA40 22396599
HeLa  Apoptosis Assay 5mM 24h induces caspase-dependent cell death 22545128
U251  Apoptosis Assay 5mM 24h increases S1-induced cell death 22579788
MCF-7  Apoptosis Assay 0.1mM 6h enhances sirtinol-induced apoptosis 22751989
PC-3  Apoptosis Assay 2mM 2h increases ORI-induced cell death 22745580
HCT116 Apoptosis Assay 5mM 24h DMSO enhances apigenin-induced cell death 24626522
U2OS Growth Inhibition Assay 10mM 24h intensifies the growth inhibition induced by Dox 24639013
A2780cp Apoptosis Assay 2.5mM 1h ddH2O enhances cisplatin-induced cell death 24817946
HepG2 Function Assay 5mM 4h increases cellular levels of HL  24713587
Microglia  Apoptosis Assay 5mM 24h decreases hypoxia-induced cell death 24818601
MDA-MB 231 Apoptosis Assay 5mM 0.5h modulates Tocomin® induced apoptosis 24830781
PANC-1  Apoptosis Assay 1mM 48h DMSO enhances bortezomib-induced cell viability loss 24842158
MDA-MB-231 Function Assay 2mM 48h promotes TM-induced cell death 24970676
MDA-MB-231 Function Assay 2mM 24h inhibits autophagy induced by TM  24970676
MCF-7  Function Assay 2mM 48h promotes TM-induced cell death 24970676
MCF-7  Function Assay 2mM 24h inhibits autophagy induced by TM  24970676
HepG2 Apoptosis Assay 3mM 5h reduces cell apoptosis induced by QDs 22836595
HeLa Apoptosis Assay 10mM 2h decreases cell viability co-treatment with PEI 23000135
SK-HEP-1 Apoptosis Assay 10mM 1h protects against autophagy and induces apoptosis in bufalin-treated cells 22858649
MDA-MB231 Function Assay 5mM 1h increases resveratrol-mediated caspase activation and cell death 23088850
PaCa44 Apoptosis Assay 2.5mM 1h reduces genipin-mediated apoptosis 23124112
T-47D Function Assay 10mM 2h inhibits autophagy process and increases rapamycin induced apoptosis 23300026
GTL-16  Apoptosis Assay 5mM 24h reduces cell viability as compared to cells treated with MET inhibitors 23313490
U251MG  Function Assay 3mM 1h suppresses LC3-II protein expression 23338618
T24 Function Assay 10mM 1h reduces the cleavage of LC3 after baicalin treatment 23354080
HUVECs Function Assay 3mM 24h blocks the protective effect of resveratrol by inhibiting autophagy 23358928
MCF-7  Function Assay 5mM 24h inhibits starvation-induced autophagy 23395679
Hela  Function Assay 5mM 24h inhibits starvation-induced autophagy 23395679
OR6 Function Assay 10mM 72h suppresses HCV replication and formation of autophagosomes 23395875
HT-29  Function Assay 1mM 48/96h inhibits AMPK induces autophagic cell death 23508272
SH-SY5Y Cytotoxicity Assay 5mM 24h increases PCN toxicity 23525265
Saos-2  Apoptosis Assay 1mM 96h increases cell death induced by PCX 23563171
1321N1 Cytotoxicity Assay 5mM 24h protects cell against PCN-induced toxicity 23525265
A2780 Apoptosis Assay 5mM 24h converts FTY720 with CDDP into an additive effect towards killing ovarian cancer cells 23592281
OV2008 Apoptosis Assay 5mM 24h converts FTY720 with CDDP into an additive effect towards killing ovarian cancer cells 23592281
PC12  Function Assay 10mM 24h water  inhibits chymotrypsin-like proteasomal activity. 23603979
SH-SY5Y Apoptosis Assay 5mM 1h abolishes celastrol neuroprotective effect 23619395
SH-SY5Y Function Assay 1mM 24h inhibits the autophagy induced by TOCP 23743148
HepG2 Function Assay 10mM 24h inhibits siTIGAR- and HBSS-induced autophagy 23817040
HeLa Function Assay 10mM 2h suppresses LC3 II expressison 23864738
HONE-1  Function Assay 5mM 1h represses 6r-mediated ROS production 23892358
MCF7 Function Assay 5mM 24h increases CuO induced cell death 23962629
HO8910 Apoptosis Assay 10mM 12h enhances B19-induced apoptosi 23983610
SMMC-7721 Apoptosis Assay 5mM 24h attenuates TNF-α protection against serum starvation-mediated apoptosis 24066693
Hep3B Apoptosis Assay 5mM 24h attenuates TNF-α protection against serum starvation-mediated apoptosis 24066693
H460 Function Assay 10mM 4h increases cisplatin-induced cell death 24173208
A549 Function Assay 10mM 4h inhibits autophagy induced by irradiation 24142735
H1299  Function Assay 10mM 4h increases cisplatin-induced cell death 24173208
WiDr Function Assay 10mM 1h inhibits PCBL-induced LC3 II expression 24190489
LoVo  Apoptosis Assay 5mM 48h enhances DCA-induced apoptosis. 24201812
HepG2 E47 Function Assay 2.5mM 48h increases the toxicity of AA, BSO, and CCl4 24273738
RKO Function Assay 2mM 1h DMSO enhances cell death by geldanamycin 24291777
Hep3B Apoptosis Assay 2mM 12h DMSO inhibits AZD8055-induced cell death 24297300
ACHN-5968 Apoptosis Assay 5mM 3h enhances paclitaxel-mediated apoptosis  24305604
Huh7 Apoptosis Assay 2mM 12h DMSO inhibits AZD8055-induced cell death 24297300
UOK257 Apoptosis Assay 5mM 3h enhances paclitaxel-mediated apoptosis  24305604
ECSCs  Apoptosis Assay 10mM 4h decreases rapamycin-treated apoptosis 24319109
MCF-7  Function Assay 10mM 24h inhibits the autophagy induced by chemotherapy drugs 24315578
SGC-7901  Apoptosis Assay 2mM 1h increases CA-4 induced apoptosis 24321340
SMMC-7721 Apoptosis Assay 2mM 1h increases CA-4 induced apoptosis 24321340
T24 Apoptosis Assay 5mM 1.5h potentiates celecoxib-induced apoptosis 24349176
NTUB1 Apoptosis Assay 5mM 1.5h potentiates celecoxib-induced apoptosis 24349176
MG-63 Apoptosis Assay 10mM 12h enhances DP-induced apoptosis 24358301
MG-63 Apoptosis Assay 0.5/1mM 32h enhances salinomycin-induced cell apoptosis 24358342
MG-63 Function Assay 0.5/1mM 48h induces salinomycin-induced cell viability loss 24358342
U2OS  Function Assay 0.5/1mM 48h induces salinomycin-induced cell viability loss 24358342
HGC-27 Function Assay 10mM 1h inhibits the cell viability loss by RAD001 or MK-2206 24416349
HCT116  Apoptosis Assay 5mM 24h enhances the apoptosis induced by apigenin 24626522
A549 Apoptosis Assay 10mM 48h accelerates the reduction of cell viability induced by PTX 24626722
Saos-2 Apoptosis Assay 10mM 24h intensifies the growth inhibition of the U2OS cells induced by Dox 24639013
U2OS Apoptosis Assay 10mM 24h intensifies the growth inhibition of the U2OS cells induced by Dox 24639013
HepG2 Function Assay 5mM 4h increases HL release 24713587
A549 Apoptosis Assay 5mM 48h decreases the percentage of cell death and expression levels of caspase-3, Beclin-1 and LC3-II 24706303
A2780cp Apoptosis Assay 2.5mM 1h ddH2O enhances cisplatin-induced cell death 24817946
Microglia  Apoptosis Assay 5mM 24h decreases hypoxia-induced cell death 24818601
HT-29  Apoptosis Assay 1mM 48h DMSO enhances bortezomib-induced cell viability loss 24842158
MDR Apoptosis Assay 10mM 6h strengthens the power of anticancer drugs 25019701
H157 Function Assay 5mM 2h suppresses SPC induced accumulation of LC3-II 25285628
A549  Function Assay 5mM 2h suppresses SPC induced accumulation of LC3-II 25285628
A2780cp  Growth Inhibition Assay 1mM 1h increases cisplatin-induced cell death 25322694
NBL-W-S  Apoptosis Assay 1mM 6h increases cell apoptosis induced by GANT-61 25323222
NBL-W-S  Growth Inhibition Assay 1mM 6h enhances GANT-61 toxicity 25323222
A549  Apoptosis Assay 5mM 2h DMSO inhibits BDMC-induced apoptotic cell death 25716561
95D Apoptosis Assay 5mM 2h DMSO inhibits BDMC-induced apoptotic cell death 25716561
A549  Growth Inhibition Assay 3mM 2h DMSO reduces growth inhibitory effect of BDMC 25716561
95D Growth Inhibition Assay 3mM 2h DMSO reduces growth inhibitory effect of BDMC 25716561
Nara-H Growth Inhibition Assay 5mM 48h enhances temsirolimusmediated suppression of Nara-H cell proliferation 21805033
HUVECs Function Assay 10mM 0.5h decreases the AGE-BSAinduced autophagy leve 21468592
HepG2 Apoptosis Assay 2mM 1h enhances radiation-induced cell death 21453691
U-2 OS qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for U-2 OS cells 29435139
A673 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells 29435139
DAOY qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for DAOY cells 29435139
Saos-2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Saos-2 cells 29435139
BT-37 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 cells 29435139
RD qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for RD cells 29435139
SK-N-SH qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-SH cells 29435139
BT-12 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-12 cells 29435139
MG 63 (6-TG R) qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for MG 63 (6-TG R) cells 29435139
NB1643 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells 29435139
OHS-50 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for OHS-50 cells 29435139
Rh41 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh41 cells 29435139
Rh30 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh30 cells 29435139
SJ-GBM2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SJ-GBM2 cells 29435139
SK-N-MC qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells 29435139
NB-EBc1 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells 29435139
LAN-5 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells 29435139
Rh18 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh18 cells 29435139
Click to View More Cell Line Experimental Data

Biological Activity

Description 3-MA (3-Methyladenine) is a selective PI3K inhibitor for Vps34 and PI3Kγ with IC50 of 25 μM and 60 μM in HeLa cells; blocks class I PI3K consistently, whereas suppression of class III PI3K is transient, and also blocks autophagosome formation. Solutions are unstable and should be fresh-prepared.
Targets
Autophagy [1] Vps34 [1]
(HeLa cells)		 PI3Kγ [1]
(HeLa cells)
25 μM 60 μM
In vitro
In vitro

The slight preference for Vps34 prevention by 3-Methyladenine probably arises from a hydrophobic ring specific to Vps34, which encircles the 3-methyl group of 3-Methyladenine. [1] 3-Methyladenine has been reported to cause cancer cell death under both normal and starvation conditions. 3-Methyladenine could also suppress cell migration and invasion independently of its ability to inhibit autophagy, implying that 3-Methyladenine possesses functions other than autophagy suppression. 3-Methyladenine elicits caspase-dependent cell death that is independent of autophagy inhibition. Treatment with 5 mM 3-Methyladenine reduces the percentage of glucose-starved HeLa cells displaying GFP-LC3 puncta to 23%. The levels of LC3-I are increasing and the levels of LC3-II are decreasing between 12 and 48 hours in cells that are treated with 3-Methyladenine. Conversion of LC3-I to LC3-II is suppressed by 3-Methyladenine. Treatment of HeLa cells with 3-Methyladenine at 2.5 mM or 5 mM for one day does not affect cell viability, whereas treatment with 10 mM 3-Methyladenine for one day causes a 25.0% decrease in cell viability. Treatment of cells with 2.5, 5 or 10 mM 3-Methyladenine for two days causes 11.5%, 38.0% and 79.4% decrease in viability, respectively. 3-Methyladenine decreases cell viability in a time- and dose-dependent manner. 3-Methyladenine significantly shortens the duration of nocodazole-induced-prometaphase arrest. [2] Suppression of autophagy by 3-Methyladenine inhibits SU11274-induced cell death. [3] Prolonged treatment with 3-Methyladenine (up to 9 hours) induces significant LC3 I to II conversion in wild type MEFs. Prolonged treatment with 3-Methyladenine, but not wortmannin, markedly increases GFP-LC3 punctuation/aggregation. 3-Methyladenine-induced LC3 conversion and free GFP liberation are ATG7-dependent. 3-Methyladenine treatment leads to evident increase of p62 protein level. 3-Methyladenine increases the p62 level even in Atg5−/− MEFs as well as in cells with DOX-mediated deletion of ATG5. 3-Methyladenine inhibits class I and class III PI3K in different temporal patterns. 3-Methyladenine-induced LC3 I to LC3 II conversion is dramatically compromised in Tsc2−/− cells compared with wild type cells.3-Methyladenine disrupts the anti-autophagic function of mTOR complex 1. [4]
Kinase Assay Protein degradation assay
HeLa cells are radiolabeled for 24 hours with 0.05 mCi/mL l-[U- 14C]valine. At the end of the labeling period, cells are rinsed three times with PBS. Cells are incubated for the designated times in either full medium or EBSS with or without the presence of 10 mM 3-Methyladenine.
Cell Research Cell lines HeLa cell line
Concentrations 1-10 mM
Incubation Time 24, 48 or 72 hours
Method

Cell (such as HeLa cell) viability is determined by a trypan blue exclusion assay. Briefly, after treated with 3-Methyladenine, both adherent and floating cells are collected and suspended in phosphate buffered saline (PBS, pH 7.4) at a final density of 1-2 × 106/mL. An equal volume of 0.4% trypan blue solution (w/v, in PBS) is added to the cell suspension and mixed thoroughly. After incubation at room temperature for 3 min, cell counting is performed using a hemacytometer.
Experimental Result Images Methods Biomarkers Images PMID
Western blot α-SMA / TGF-β / LC-3BI / LC-3B II / Beclin-1 / NF-κB p65 caspase-3 / caspase-9 / PARP VEGF APP / BACE1 / ADAM17 / Presenilin 1 / Presenilin 2 / Nicastrin / APH-1 / Pen-2 / LC3-1 / LC3-2 29296191
Immunofluorescence LC3 / Hif-α / COX2 29039446
Growth inhibition assay Cell viability 26934124
In Vivo
In vivo

3-Methyladenine blocks autophagy through its effect on class III phosphatidylinositol 3-kinase (PI3K). 3-Methyladenine treatment does not alter the degree of hemorrhage compared with the subarachnoid hemorrhage (SAH) group. 3-Methyladenine pretreatment significantly aggravates neurological symptoms when compared with the SAH + vehicle group. Autophagy is decreased when 3-Methyladenine treatment is applied. Conversely, cleaved caspase-3 is markedly up-regulated in the SAH + 3-Methyladenine group. In line with the up-regulation of cleaved caspase-3 expression, the number of TUNEL-positive cells in the right cortex is significantly increased in the SAH + 3-Methyladenine group compared with the SAH + vehicle group. [5]
Animal Research Animal Models Adult male Sprague–Dawley rats weighing 300-350 g
Dosages 400 nM
Administration Intracerebral ventricular

Chemical Information & Solubility

Molecular Weight 149.15 Formula

C6H7N5
CAS No. 5142-23-4 SDF Download 3-MA (3-Methyladenine) SDF
Smiles CN1C=NC(=N)C2=C1N=CN2
Storage (From the date of receipt) 3 years -20°C powder Solutions are unstable. Prepare fresh or purchase small, pre-packaged sizes. Repackage upon receipt.

In vitro
Batch:

DMSO : 10 mg/mL ( (67.04 mM) Warmed with 50°C water bath; Ultrasonicated; Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Ethanol : 10 mg/mL

Water : 4 mg/mL


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In vivo
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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

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Frequently Asked Questions

Question 1:
I'm also wondering whether it can be dissolved in water,or maybe something like culture medium,normal saline solution to form 10mM solution

Answer:
As the reference (http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal. pone.0035665), 3-MA, which was found to inhibit autophagy at concentrations ranging from 1 to 10 mM was directly dissolved into the culture medium at the indicated concentrations. And we tested the solubility of S2767, and found the solubility of 3-MA in DMEM is 31 mg/mL at about 40°C.

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