| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 1g | |||
| Other Sizes |
| Targets |
MTX/methotrexate analog; DHFR/dihydrofolate reductase; anticancer; FPGS/folylpoly-γ-glutamate synthetase
The primary targets of NSC269401 include dihydrofolate reductase (DHFR) and folylpoly-γ-glutamate synthetase (FPGS). It acts as a DHFR inhibitor and a substrate for FPGS. Methotrexate Diglutamate is a metabolite of methotrexate and retains the ability to inhibit DHFR, contributing to the pharmacological effects of methotrexate. |
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| ln Vitro |
Ester prodrugs of the phosphinate pseudopeptide N-[(4-deoxy-4-amino-10-methyl)pteroyl]glutamate-gamma-[psiP(O)(OH)]-glutarate (1a) were synthesized. H-phosphinic acids derived from N-Cbz vinyl glycine esters were converted to the desired pseudopeptides by Michael addition to alpha-methyleneglutarate esters. Pivaloyloxymethyl (POM) ester moieties were incorporated in both the N-terminal and C-terminal fragments prior to formation of either C-P bond. N-Alkylation of the corresponding amides derived from p-(N-methyl)aminobenzoic acid with 2,4-diamino-6-(bromomethyl)pteridine gave the target compounds. POM esters of methotrexate and the corresponding gamma-glutamyl conjugate were also synthesized using the same strategy. All prodrugs were evaluated in Chinese hamster ovary cells. Although the pseudopeptide prodrugs were ineffective, prodrugs of methotrexate and the corresponding gamma-glutamyl conjugate were equipotent with the parent compounds. Stability of the prodrugs was investigated in both phosphate buffer and cell line medium to provide a rationale for the observed biological data [1].
In vitro, NSC269401 is a DHFR inhibitor. It is a small molecule compound present in the liver that acts as a partially purified human dihydrofolate reductase inhibitor. AMPte-Glu-γ-Glu has been shown to be cytotoxic to Chinese hamster ovary cells, with IC50 values of 20 nM for the parental cell line and higher values for derived cell lines. These assays confirm that NSC269401 and related compounds have anti-proliferative activity. |
| ln Vivo |
In vivo, NSC269401 is a metabolite of methotrexate and is present in the liver. As a DHFR inhibitor, it contributes to the pharmacological effects of methotrexate, including its antineoplastic and antirheumatic activities.
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| Enzyme Assay |
In vitro enzyme/receptor binding assays for NSC269401 involve DHFR inhibition assays using purified DHFR enzyme. The compound's inhibitory activity is measured by monitoring the reduction of dihydrofolate to tetrahydrofolate in the presence of NADPH. IC50 values are determined from dose-response curves. FPGS assays can be performed to assess the compound's activity as a substrate for this enzyme.
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| Cell Assay |
In vitro cellular assays for NSC269401 are conducted in cancer cell lines. Cells are treated with varying concentrations of NSC269401, and cell proliferation is measured using MTT, CellTiter-Glo, or colony formation assays. Cytotoxicity is assessed by measuring cell viability. These assays confirm that NSC269401 has anti-proliferative activity.
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| Animal Protocol |
In vivo animal studies for NSC269401 are not typically performed as a standalone compound, as it is a metabolite of methotrexate. However, its effects would be similar to those of methotrexate, which is studied in animal models of cancer and autoimmune diseases.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of NSC269401 indicate that it has a molecular weight of 583.55 and a molecular formula of C25H29N9O8. As a metabolite of methotrexate, its pharmacokinetics are related to those of the parent drug. The compound is typically stored as a powder at appropriate conditions to maintain stability.
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| Toxicity/Toxicokinetics |
The toxicological profile of NSC269401 is similar to that of methotrexate, which can cause myelosuppression, hepatotoxicity, and gastrointestinal toxicity. As a DHFR inhibitor, it can affect rapidly dividing cells, leading to these side effects.
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| References |
[1]. Prodrug forms of N-[(4-deoxy-4-amino-10-methyl)pteroyl]glutamate-gamma-[psiP(O)(OH)]-glutarate, a potent inhibitor of folylpoly-gamma-glutamate synthetase: synthesis and hydrolytic stability. J Med Chem. 2006 Jan 26;49(2):770-88.
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| Additional Infomation |
NSC269401 is a metabolite of methotrexate and a DHFR inhibitor. It is also known as Methotrexate Diglutamate. The compound is a substrate for FPGS. NSC269401 is not approved for clinical use and is available from research chemical suppliers for research applications.
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| Molecular Formula |
C25H29N9O8
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|---|---|
| Molecular Weight |
583.56
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| Exact Mass |
583.214
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| Elemental Analysis |
C, 51.46; H, 5.01; N, 21.60; O, 21.93
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| CAS # |
41600-13-9
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| PubChem CID |
430166
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| Appearance |
Typically exists as solid at room temperature
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| Density |
1.539g/cm3
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| Index of Refraction |
1.704
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| LogP |
1.562
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| Hydrogen Bond Donor Count |
7
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| Hydrogen Bond Acceptor Count |
15
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| Rotatable Bond Count |
14
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| Heavy Atom Count |
42
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| Complexity |
980
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CN(CC1=CN=C2C(=N1)C(=NC(=N2)N)N)C3=CC=C(C=C3)C(=O)NC(CCC(=O)NC(CCC(=O)O)C(=O)O)C(=O)O
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| InChi Key |
PIHMDTSRKDVWKE-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C25H29N9O8/c1-34(11-13-10-28-21-19(29-13)20(26)32-25(27)33-21)14-4-2-12(3-5-14)22(38)31-16(24(41)42)6-8-17(35)30-15(23(39)40)7-9-18(36)37/h2-5,10,15-16H,6-9,11H2,1H3,(H,30,35)(H,31,38)(H,36,37)(H,39,40)(H,41,42)(H4,26,27,28,32,33)
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| Chemical Name |
2-[[4-carboxy-4-[[4-[(2,4-diaminopteridin-6-yl)methyl-methylamino]benzoyl]amino]butanoyl]amino]pentanedioic acid
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| Synonyms |
NSC-269401; NSC 269401; NSC269401; 41600-13-9; 4-Amino-4-deoxy-N10-methylpteroylglutamyl-g-glutamate; NSC 269401; 2-[[4-carboxy-4-[[4-[(2,4-diaminopteridin-6-yl)methyl-methylamino]benzoyl]amino]butanoyl]amino]pentanedioic acid; L-Glutamic acid, N-(4-(((2,4-diamino-6-pteridinyl)methyl)methylamino)-; L-Glutamic acid, N-(4-(((2,4-diamino-6-pteridinyl)methyl)methylamino)benzoyl)-L-g-glutamyl-; (2S)-2-[[(4S)-4-carboxy-4-[[4-[(2,4-diaminopteridin-6-yl)methyl-methylamino]benzoyl]amino]butanoyl]amino]pentanedioic acid; NSC269401
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7136 mL | 8.5681 mL | 17.1362 mL | |
| 5 mM | 0.3427 mL | 1.7136 mL | 3.4272 mL | |
| 10 mM | 0.1714 mL | 0.8568 mL | 1.7136 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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