Synonym
Methotrexate hydrate; Amethopterin; CL 14,377; EMT 25299; MTX; NSC 740
IUPAC/Chemical Name
(4-(((2,4-diaminopteridin-6-yl)methyl)(methyl)amino)benzoyl)-L-glutamic acid hydrate
InChi Key
FPJYMUQSRFJSEW-ZOWNYOTGSA-N
InChi Code
InChI=1S/C20H22N8O5.H2O/c1-28(9-11-8-23-17-15(24-11)16(21)26-20(22)27-17)12-4-2-10(3-5-12)18(31)25-13(19(32)33)6-7-14(29)30;/h2-5,8,13H,6-7,9H2,1H3,(H,25,31)(H,29,30)(H,32,33)(H4,21,22,23,26,27);1H2/t13-;/m0./s1
SMILES Code
O=C(O)CC[C@@H](C(O)=O)NC(C1=CC=C(N(CC2=NC3=C(N)N=C(N)N=C3N=C2)C)C=C1)=O.[H]O[H]
Purity
>98% (or refer to the Certificate of Analysis)
Shipping Condition
Shipped under ambient temperature as non-hazardous chemical. This product is stable enough for a few weeks during ordinary shipping and time spent in Customs.
Storage Condition
Dry, dark and at 0 - 4 C for short term (days to weeks) or -20 C for long term (months to years).
Solubility
Soluble in DMSO
Shelf Life
>3 years if stored properly
Drug Formulation
This drug may be formulated in DMSO
Stock Solution Storage
0 - 4 C for short term (days to weeks), or -20 C for long term (months).
HS Tariff Code
2934.99.9001
Biological target:
Methotrexate hydrate acts by inhibiting the metabolism of folic acid and blocking key enzymes in the synthesis of purines and pyrimidines required for cell proliferation.
In vitro activity:
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. 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.
Reference: J Clin Invest. 1998 Jul 15;102(2):322-8. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC508890/
Arthritis Res. 2002;4(4):266-73. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC128935/
In vivo activity:
Methotrexate reduces thymus and spleen indices of mice. Methotrexate markedly decreases white blood cells, thymic and splenic lymphocytes at dose greater than or equal to 5 mg/kg. However, there is a significant difference between the treatment plus control group and the model group (p<0.01). The combination of grape seed proanthocyanidins and Siberian ginseng eleutherosides obviously diminishes the effects of Methotrexate exposure on indices of thymus and spleens in mice
Reference: Pharmacol Rep. 2006 Jul-Aug;58(4):473-92. http://if-pan.krakow.pl/pjp/pdf/2006/4_473.pdf
|
Solvent |
mg/mL |
mM |
comments |
| Solubility |
| DMF |
14.0 |
29.63 |
|
| DMSO |
3.0 |
6.35 |
|
| PBS (pH 7.2) |
1.0 |
2.12 |
|
Note: There can be variations in solubility for the same chemical from different vendors or different batches from the same vendor. The following factors can affect the solubility of the same chemical: solvent used for crystallization, residual solvent content, polymorphism, salt versus free form, degree of hydration, solvent temperature. Please use the solubility data as a reference only. Warming and sonication will facilitate dissolving. Still have questions? Please contact our Technical Support scientists.
Preparing Stock Solutions
The following data is based on the
product
molecular weight
472.46
Batch specific molecular weights may vary
from batch to batch
due to the degree of hydration, which will
affect the solvent
volumes required to prepare stock solutions.
| Concentration / Solvent Volume / Mass |
1 mg |
5 mg |
10 mg |
| 1 mM |
1.15 mL |
5.76 mL |
11.51 mL |
| 5 mM |
0.23 mL |
1.15 mL |
2.3 mL |
| 10 mM |
0.12 mL |
0.58 mL |
1.15 mL |
| 50 mM |
0.02 mL |
0.12 mL |
0.23 mL |
Formulation protocol:
1. Karasik A, Váradi A, Szeri F. In vitro transport of methotrexate by Drosophila Multidrug Resistance-associated Protein. PLoS One. 2018 Oct 12;13(10):e0205657. doi: 10.1371/journal.pone.0205657. PMID: 30312334; PMCID: PMC6185855.
2. Yang J, Gao L, Yu P, Kosgey JC, Jia L, Fang Y, Xiong J, Zhang F. In vitro synergy of azole antifungals and methotrexate against Candida albicans. Life Sci. 2019 Oct 15;235:116827. doi: 10.1016/j.lfs.2019.116827. Epub 2019 Aug 31. PMID: 31479680.
3. Previtali V, Petrovic K, Peiró Cadahía J, Troelsen NS, Clausen MH. Auxiliary in vitro and in vivo biological evaluation of hydrogen peroxide sensitive prodrugs of methotrexate and aminopterin for the treatment of rheumatoid arthritis. Bioorg Med Chem. 2020 Jan 15;28(2):115247. doi: 10.1016/j.bmc.2019.115247. Epub 2019 Dec 6. PMID: 31843461.
4. Choi SJ, Oh JM, Chung HE, Hong SH, Kim IH, Choy JH. In vivo anticancer activity of methotrexate-loaded layered double hydroxide nanoparticles. Curr Pharm Des. 2013;19(41):7196-202. doi: 10.2174/138161281941131219123718. PMID: 23489199.
In vitro protocol:
1. Karasik A, Váradi A, Szeri F. In vitro transport of methotrexate by Drosophila Multidrug Resistance-associated Protein. PLoS One. 2018 Oct 12;13(10):e0205657. doi: 10.1371/journal.pone.0205657. PMID: 30312334; PMCID: PMC6185855.
2. Yang J, Gao L, Yu P, Kosgey JC, Jia L, Fang Y, Xiong J, Zhang F. In vitro synergy of azole antifungals and methotrexate against Candida albicans. Life Sci. 2019 Oct 15;235:116827. doi: 10.1016/j.lfs.2019.116827. Epub 2019 Aug 31. PMID: 31479680.
In vivo protocol:
1. Previtali V, Petrovic K, Peiró Cadahía J, Troelsen NS, Clausen MH. Auxiliary in vitro and in vivo biological evaluation of hydrogen peroxide sensitive prodrugs of methotrexate and aminopterin for the treatment of rheumatoid arthritis. Bioorg Med Chem. 2020 Jan 15;28(2):115247. doi: 10.1016/j.bmc.2019.115247. Epub 2019 Dec 6. PMID: 31843461.
2. Choi SJ, Oh JM, Chung HE, Hong SH, Kim IH, Choy JH. In vivo anticancer activity of methotrexate-loaded layered double hydroxide nanoparticles. Curr Pharm Des. 2013;19(41):7196-202. doi: 10.2174/138161281941131219123718. PMID: 23489199.
1. Tian, H., and Cronstein, B.N. Understanding the mechanisms of action of methotrexate: Implications for the treatment of rheumatoid arthritis. Bull. NYU Hosp. Jt. Dis. 65(3), 168-173 (2007).
2. Swierkot, J., and Szechinski, J. Methotrexate in rheumatoid arthritis. Pharmacol. Rep. 58(4), 473-492 (2006).
3. Chan, E.S.L., and Cronstein, B.N. Molecular action of methotrexate in inflammatory diseases. Arthritis Res. 4(4), 266-273 (2002).
4. Wessels, J.A.M., Huizinga, T.W.J., and Guchelaar, H.J. Recent insights in the pharmacological actions of methotrexate in the treatment of rheumatoid arthritis. Rheumatology 47, 249-255 (2008).
5. Cutolo, M., Sulli, A., Pizzorni, C., et al. Anti-inflammatory mechanisms of methotrexate in rheumatoid arthritis. Ann. Rheum. Dis. 60(8), 729-735 (2001).
6. Christin-Maitre, S., Bouchard, P., and Spitz, I.M. Medical termination of pregnancy. N. Engl. J. Med. 342(13), 946-956 (2000).
7. Smolen, J.S., and Steiner, G. Therapeutic strategies for rheumatoid arthritis. Nat. Rev. Drug Dis. 2(6), 473-488 (2003).
