Catalog No.S1210 Synonyms: NCI-C04671

Methotrexate Chemical Structure

Molecular Weight(MW): 454.44

Methotrexate (MTX), analog of folic acid, is a nonspecific inhibitor of the dihydrofolate reductase(DHFR) of bacteria and cancerous cells as well as normal cells. It forms an inactive ternary complex with DHFR and NADPH.

Size Price Stock Quantity  
In DMSO USD 134 In stock
USD 97 In stock
USD 197 In stock
USD 970 In stock
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2 Customer Reviews

  • A. Viability comparison of REH vector control, BCL6 overexpression, or BCL6 overexpression cells pre-treated with 79-6 (125μM) following exposure to three chemotherapy drugs (Ara-C [1 μM], MTX [50 μM], VCR [25 μM]). (* = p < 0.05 BCL6 OX to vector control and # = p < 0.05 BCL6 OX to BCL6 + 79-6).

    Oncotarget, 2016, 7(17):23439-53. Methotrexate purchased from Selleck.

    Toxicity of MTX (3) and toxic analogues 5 a-e on a) hY1R-expressing MDA-MB-468 and b) non-hYR-expressing HEK293 cells. Cell viability was determined by resazurin assay. Bars represent the mean±SEM of at least three independent experiments performed in triplicate. Measurements were normalized by using only HBSS, pH 7, treated cells (set at 100 %) and ethanol-treated cells (set at 0 %). Statistical significances refer to only HBSS, pH 7, treated cells indicated as a dashed line.

    ChemMedChem, 2015, 10(5):804-14.. Methotrexate purchased from Selleck.

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Biological Activity

Description Methotrexate (MTX), analog of folic acid, is a nonspecific inhibitor of the dihydrofolate reductase(DHFR) of bacteria and cancerous cells as well as normal cells. It forms an inactive ternary complex with DHFR and NADPH.
hDHFR [4]
(Activated peripheral T cells)
24 nM
In vitro

Methotrexate (0.1-10 mM) induces apoptosis of in vitro activated T cells from human peripheral blood. Methotrexate achieves clonal deletion of activated T cells in mixed lymphocyte reactions. Methotrexate can selectively delete activated peripheral blood T cells by a CD95-independent pathway. [1] Methotrexate is taken up by cells via the reduced folate carrier and then is converted within the cells to polyglutamates. Methotrexate leads to diminished production of leukotriene B4 by neutrophils stimulated ex vivo. Methotrexate polyglutamates inhibit the enzyme aminoimidazolecarboxamidoadenosineribonucleotide (AICAR) transformylase more potently than the other enzymes involved in purine biosynthesis. Methotrexate is also known to suppress TNF activity by suppressing TNF-induced nuclear factor-κB activation in vitro, in part related to a reduction in the degradation and inactivation of an inhibitor of this factor, IκBα, and probably related to the release of adenosine. Methotrexate suppresses the production of both TNF and IFN-γ by T-cell-receptor-primed T lymphocytes from both healthy human donors and RA patients. Methotrexate treatment is associated with a significant decrease of TNF-α-positive CD4+ T cells, while the number of T cells expressing the anti-inflammatory cytokine IL-10 increased. [2]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
P388D1 M2LN[IN6fG:2b4jpZ4l1gSCjc4PhfS=> M1GwdmlEPTB;ND64JI5O M{PwdVg3OzJ2MUO=
L cells MXLjfZRwfG:6aXPpeJkh[XO|YYm= M4XDfmlEPTB;OD6zJI5O MXK4OlMzPDF|
D54 NVS5VpRN[3m2b4TvfIlkcXS7IHHzd4F6 MlfOTWM2OD1zNjDuUS=> MlXTPFY{OjRzMx?=
143B(TK-) M1nTVoN6fG:2b4jpZ4l1gSCjc4PhfS=> Mn\qTWM2OD16Lkigcm0> MnqyPFY{OjRzMx?=
U87MG NHrpZldkgXSxdH;4bYNqfHliYYPzZZk> M3;GfWlEPTB;MkKgcm0> M12yd|g3OzJ2MUO=
A549 M4\1ToN6fG:2b4jpZ4l1gSCjc4PhfS=> MnjBTWM2OD1|MTDuUS=> MU[4OlMzPDF|
H460 MVfjfZRwfG:6aXPpeJkh[XO|YYm= NH;qOoVKSzVyPUmuOUBvVQ>? MWC4OlMzPDF|
Daoy NWPXOZVG[3m2b4TvfIlkcXS7IHHzd4F6 NFGzd2JKSzVyPUmgcm0> M4TrXFg3OzJ2MUO=
U373MG M4\tPIN6fG:2b4jpZ4l1gSCjc4PhfS=> NXTybVc1UUN3ME2xNkBvVQ>? MkjrPFY{OjRzMx?=
Vero NGfCenlkgXSxdH;4bYNqfHliYYPzZZk> NHHsO3ZKSzVyPUmuNkBvVQ>? NX;aNYtTQDZ|MkSxNy=>
SCC25  MlrsS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? M1njNp4yKM7:TR?= NWLycXd2UUN3ME2yO{BvVQ>? MlL5PVAzOjd7NR?=
NCI-H460 NInQ[4dIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= MoO5glEh|ryP MYPJR|UxRTJ6IH7N NGP5dVk6ODJ{N{m1
NCI-H23 MXHHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NG\JTXh,OSEQvF2= MlLDTWM2OD12MzDuUS=> MmfsPVAzOjd7NR?=
NCI-H522 MV;Hdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? M1HzWJ4yKM7:TR?= NV:3UVVsUUN3ME2yNlkhdk1? NWDvZ|lrQTB{Mke5OS=>
EKVX NGXOd5VIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= NW[ycZVMhjFizszN MYHJR|UxRjFyMECgcm0> NIPvfHI6ODJ{N{m1
HCT-116 MYPHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? M2jXfZ4yKM7:TR?= M130fGlEPTB;M{Cgcm0> MV65NFIzPzl3
HCT-15 MVLHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NVr1e2F{hjFizszN M4nk[WlEPTB;M{Cgcm0> MY[5NFIzPzl3
HT29 M3fhfmdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 M2HURZ4yKM7:TR?= MmPHTWM2OD1|MjDuUS=> NIO4VZk6ODJ{N{m1
SW-620 M{GwSGdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 Mn\rglEh|ryP NV7XO2JMUUN3ME2zN{BvVQ>? M3u2PVkxOjJ5OUW=
KM12 NUPHWohKT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= MoLvglEh|ryP NEniVopKSzVyPUSyJI5O M1rZZ|kxOjJ5OUW=
SF-539 M2XTfmdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MnTkglEh|ryP NV3FTmdxUUN3ME2zOUBvVQ>? MXi5NFIzPzl3
SF-268 NXLsNWN5T3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NF36NlJ,OSEQvF2= NUXSem9CUUN3ME21NkBvVQ>? M3TlVVkxOjJ5OUW=
SNB-75 NWruV|lyT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NFfITmx,OSEQvF2= NFL1dVlKSzVyPkGwNFAxKG6P M3rHWFkxOjJ5OUW=
LOX IMVI Mk\MS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? M{HI[J4yKM7:TR?= M3Tzb2lEPTB;Mk[gcm0> MkTqPVAzOjd7NR?=
SK-MEL-5 NW\n[mNOT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NYDEdFBVhjFizszN MoT4TWM2OD16NzDuUS=> MYK5NFIzPzl3
SK-MEL-28 M1G5VGdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 M4jmcp4yKM7:TR?= MXHJR|UxRjFyMECgcm0> NEOwSYQ6ODJ{N{m1
OVCAR-8 MWfHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? M3PmUp4yKM7:TR?= M1y1RmlEPTB;M{Ggcm0> MUK5NFIzPzl3
OVCAR-5 Mn\DS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? NW\wV4xNhjFizszN NFnCUGdKSzVyPkGwNFAhdk1? NFjpfmo6ODJ{N{m1
OVCAR-3 M2HHeGdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 M4rNTJ4yKM7:TR?= M{TyV2lEPTB;M{m4JI5O MVO5NFIzPzl3
786-0 NUDmTGdyT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NYXBb|BHhjFizszN Mmm3TWM2OD1|MzDuUS=> NHvmVZg6ODJ{N{m1
UO-31 M3zIVWdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 NUDySYNvhjFizszN MlXWTWM2OD1zOUGgcm0> NYXnSWZHQTB{Mke5OS=>
ACHN MmC2S5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? MnP0glEh|ryP Mk\zTWM2OD12MDDuUS=> M3HLeFkxOjJ5OUW=
MCF7 NV3KbWZxT3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NX\hZmRihjFizszN NWPqOZhHUUN3ME2zOkBvVQ>? NXfYN2gzQTB{Mke5OS=>
MCF7-ADR MWHHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? MXj+NUDPxE1? M33LW2lEPTB;N{igcm0> NFf0dWk6ODJ{N{m1
PC-3 MXzHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NF7PWGx,OSEQvF2= NWHhbXdbUUN3ME2yJI5O M{\BOlkxOjJ5OUW=
DU-145 MmLtS5Jwf3SqIHnubIljcXSxcomgZZN{[Xl? M3;5dZ4yKM7:TR?= NWLiXYQyUUN3ME2yN{BvVQ>? MmLrPVAzOjd7NR?=
CCRF-CEM MVrHdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? NFrqeZlGSzVyPUG0MlQhdk1? M1PPV|ExQTV4MkKx
R1 NF\ufWVIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= M1y4VmVEPTB;Nke1JI5O MWixNFk2PjJ{MR?=
R2(Bos) NFnpXZJIem:5dHigbY5pcWKrdH;yfUBie3OjeR?= MWjFR|UxRTF4MECgcm0> Mlj5NVA6PTZ{MkG=
R30dm M{ThSWdzd3e2aDDpcohq[mm2b4L5JIF{e2G7 MnXJSWM2OD1zNDDuUS=> MWexNFk2PjJ{MR?=
FaDu MV;Hdo94fGhiaX7obYJqfG:{eTDhd5NigQ>? MkTVSWM2OD1zMT6zJI5O NHHGW5kyODl3NkKyNS=>
A253 NV\OVnA1T3Kxd4ToJIlvcGmkaYTvdpkh[XO|YYm= NET4UnZGSzVyPUG0MlUhdk1? NGLUSWIyODl3NkKyNS=>
HT-29 M37iOYN6fG:2b4jpZ4l1gSCjc4PhfS=> MXLJR|UxRTNwNEWg{txO NFXOe2wzOzl4OEiyOC=>
COLO-320 DM NVPEOWFb[3m2b4TvfIlkcXS7IHHzd4F6 NEXkdWdKSzVyPUWuNlUh|ryP NF3KXmozOzl4OEiyOC=>
COLO 205 MUHjfZRwfG:6aXPpeJkh[XO|YYm= NXT6clRxUUN3ME2zMlI4KM7:TR?= NV3PUoVSOjN7Nki4NlQ>
BGC-823 M1P2ZoN6fG:2b4jpZ4l1gSCjc4PhfS=> NVm2fIxXUUN3ME2wMlEyKM7:TR?= M3vBV|E5PTV3NU[y
Hela MkP3Z5l1d3SxeHnjbZR6KGG|c3H5 MVXJR|UxRTBwMTFOwG0> MVmxPFU2PTV4Mh?=
Bel-7402 Mk[4Z5l1d3SxeHnjbZR6KGG|c3H5 MmDVTWM2OD16Nz65JO69VQ>? MVixPFU2PTV4Mh?=

... Click to View More Cell Line Experimental Data

In vivo Methotrexate increases splenocyte AICAR content, raised adenosine concentrations in exudates from carrageenan-inflamed air pouches, and markedly inhibits leukocyte accumulation in inflamed air pouches in mice. Methotrexate-mediated reduction in leukocyte accumulation is partially reversed by injection of adenosine deaminase (ADA) into the air pouch, completely reverses by a specific adenosine A2 receptor antagonist, 3,7-dimethyl-1-propargylxanthine (DMPX), but not affected by an adenosine A1 receptor antagonist, 8-cyclopentyl-dipropylxanthine in mice. [3]


Solubility (25°C)

In vitro DMSO 90 mg/mL warmed (198.04 mM)
Water Insoluble
Ethanol Insoluble
In vivo Add solvents individually and in order:
2% DMSO+30% PEG 300+5% Tween 80+ddH2O

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 454.44


CAS No. 59-05-2
Storage powder
Synonyms NCI-C04671

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Clinical Trial Information

NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT01338987 Active, not recruiting Myelodysplastic Syndrome RAEB 2|Acute Lymphocytic Leukemia|Acute Myelogenous Leukemia|Myelodysplastic Syndrome RAEB 1 National Cancer Institute (NCI)|National Institutes of Health Clinical Center (CC) March 30, 2011 Phase 2
NCT02250937 Recruiting Acute Myeloid Leukemia|Myelodysplastic Syndrome M.D. Anderson Cancer Center October 27, 2014 Phase 2
NCT01875237 Active, not recruiting Leukemia|Myeloma|Myeloproliferative Diseases M.D. Anderson Cancer Center|Bellicum Pharmaceuticals December 27, 2013 Phase 1|Phase 2
NCT02385110 Recruiting Leukemia M.D. Anderson Cancer Center September 23, 2015 Phase 2
NCT03007147 Not yet recruiting B Acute Lymphoblastic Leukemia With t(9;22)(q34.1;q11.2); BCR-ABL1|BCR-ABL1 Fusion Protein Expression|Minimal Residual Disease|Philadelphia Chromosome Positive|T Acute Lymphoblastic Leukemia|Untreated Adult Acute Lymphoblastic Leukemia|Untreated Childhood Acute Lymphoblastic Leukemia Childrens Oncology Group|National Cancer Institute (NCI) July 2017 Phase 3
NCT02724904 Not yet recruiting Lymphoma Massachusetts General Hospital|Adienne SA|Dana-Farber Cancer Institute May 2017 --

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DHFR Signaling Pathway Map

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID