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Type HazMat fee for 500 gram (Estimated)
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Chemical Structure| 59-05-2 Chemical Structure| 59-05-2
Chemical Structure| 59-05-2

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Methotrexate, an antifolate compound, 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 and can interfere with the growth of certain cells of the body, especially cells that reproduce quickly, such as cancer cells, bone marrow cells, and skin cells.

Synonyms: Amethopterin; CL14377; NCI-C04671

4.5 *For Research Use Only !

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Product Citations

Product Citations

Verónica Urdaneta-Páez ; Randy Hamchand ; Karen Anthony ; Jason Crawford ; Alan G. Sutherland ; Barbara I. Kazmierczak

Abstract: Pseudomonas aeruginosa is intrinsically resistant to many classes of antibiotics, reflecting the restrictive nature of its outer membrane and the action of its numerous efflux systems. However, the dynamics of compound uptake, retention, and efflux in this bacterium remain incompletely understood. Here, we exploited the sensor capabilities of a Z-nucleotide-sensing riboswitch to create an experimental system able to identify physicochemical and structural properties of compounds that permeate the bacterial cell, avoid efflux, and perturb the folate cycle or de novo purine synthesis. In the first step, a collection of structurally diverse compounds enriched in antifolate drugs was screened for ZTP (5-aminoimidazole-4-carboxamide riboside 5′-triphosphate) riboswitch reporter activity in efflux-deficient P. aeruginosa, allowing us to identify compounds that entered the cell and disrupted the folate pathway. These initial hits were then rescreened using isogenic efflux-proficient bacteria, allowing us to separate efflux substrates from efflux avoiders. We confirmed this categorization by measuring intracellular levels of select compounds in the efflux-deficient and -proficient strain using high-resolution liquid chromatography-mass spectrometry (LC-MS). This simple yet powerful method, optimized for high-throughput screening, enables the discovery of numerous permeable compounds that avoid efflux and paves the way for further refinement of the physicochemical and structural rules governing efflux in this multidrug-resistant Gram-negative pathogen. IMPORTANCE: Treatment of Pseudomonas aeruginosa infections has become increasingly challenging. The development of novel antibiotics against this multidrug-resistant bacterium is a priority, but many drug candidates never achieve effective concentrations in the bacterial cell due to its highly restrictive outer membrane and the action of multiple efflux pumps. Here, we develop a robust and simple reporter system in P. aeruginosa to screen chemical libraries and identify compounds that either enter the cell and remain inside or enter the cell and are exported by efflux systems. This approach enables the development of rules of compound uptake and retention in P. aeruginosa that will lead to more rational design of novel antibiotics.

Keywords: Pseudomonas aeruginosa ; antimicrobial resistance ; drug efflux

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Product Details of Methotrexate

CAS No. :59-05-2
Formula : C20H22N8O5
M.W : 454.44
SMILES Code : O=C(O)[C@@H](NC(C1=CC=C(N(CC2=NC3=C(N)N=C(N)N=C3N=C2)C)C=C1)=O)CCC(O)=O
Synonyms :
Amethopterin; CL14377; NCI-C04671
MDL No. :MFCD00064370
InChI Key :FBOZXECLQNJBKD-ZDUSSCGKSA-N
Pubchem ID :126941

Safety of Methotrexate

GHS Pictogram:
Signal Word:Danger
Hazard Statements:H301-H341-H360-H372
Precautionary Statements:P202-P260-P264-P280-P301+P310-P405
Class:6.1
UN#:1544
Packing Group:

Application In Synthesis of Methotrexate

* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.

  • Downstream synthetic route of [ 59-05-2 ]

[ 59-05-2 ] Synthesis Path-Downstream   1~2

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  • [ 59-05-2 ]
  • [ 945-24-4 ]
  • [ 52980-68-4 ]
  • 2
  • [ 1035373-85-3 ]
  • [ 59-05-2 ]
  • [ 1613616-96-8 ]
YieldReaction ConditionsOperation in experiment
With 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; triethylamine; In N,N-dimethyl-formamide; for 3h; 50 mg of methotrexate hydrate was stirred in 3 mL of DMF and treated with ED AC (63 mg, 330 umol) and triethylamine (77 uL, 550 umol). After 10 min, 12-chloro-3, 6-dioxo- dodecylamine hydrocholide (21.5 mg, 83 umol) was added. After 3 h, the product was isolated by preparative HPLC (2->50% MeCN in 0.1% aqueous formic acid). The appropriate fractions were concentrated and lyophilized to yield an orange solid. Calculated for M+H: 660.3; found 660.7
 

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