| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 500mg |
|
||
| Other Sizes |
| Targets |
Dihydrofolate targets dihydrofolate reductase (DHFR), the enzyme responsible for its reduction to tetrahydrofolate. DHFR catalyzes the NADPH-dependent reduction of dihydrofolate to tetrahydrofolate, a critical cofactor required for the synthesis of purines, thymidylate, and several amino acids. Dihydrofolate is the natural substrate for DHFR and is also a direct product of thymidylate synthase.
|
|---|---|
| ln Vitro |
In vitro, dihydrofolate is used as a substrate to measure DHFR activity in enzyme assays. Its conversion to tetrahydrofolate by DHFR can be monitored spectrophotometrically or by HPLC. Dihydrofolate is also used in studies of folate metabolism and as a reference standard for the quantification of folate metabolites in biological samples.
|
| ln Vivo |
In vivo, dihydrofolate is a metabolic intermediate in the folate pathway. It is produced by the action of thymidylate synthase on deoxyuridylate and is rapidly reduced to tetrahydrofolate by DHFR. Disruption of dihydrofolate metabolism is a key mechanism of action for antifolate drugs such as methotrexate, which target DHFR to block DNA synthesis.
|
| Enzyme Assay |
In vitro enzyme assays for dihydrofolate measure the activity of dihydrofolate reductase (DHFR). The enzyme is incubated with dihydrofolate substrate and NADPH, and the production of tetrahydrofolate is measured spectrophotometrically at 340 nm. The decrease in absorbance at 340 nm due to NADPH oxidation is monitored over time. IC50 values for DHFR inhibitors are calculated from inhibition curves.
|
| Cell Assay |
In vitro cell-based assays using dihydrofolate assess cellular DHFR activity and folate metabolism. Cells are cultured in folate-deficient media, and the conversion of dihydrofolate to tetrahydrofolate is measured. Antifolate drugs such as methotrexate are evaluated for their ability to inhibit cellular DHFR activity by measuring the accumulation of dihydrofolate or the depletion of tetrahydrofolate.
|
| Animal Protocol |
In vivo animal models for dihydrofolate metabolism include models of folate deficiency and antifolate drug toxicity. Rodents are treated with DHFR inhibitors such as methotrexate, and the effects on dihydrofolate accumulation, tetrahydrofolate depletion, and hematopoietic toxicity are evaluated. These models are used to study the pharmacology and toxicology of antifolate drugs.
|
| ADME/Pharmacokinetics |
Pharmacokinetic properties of dihydrofolate are not well characterized as it is a metabolic intermediate rather than a drug candidate. Dihydrofolate is produced endogenously and is rapidly metabolized by DHFR. In cells treated with DHFR inhibitors, dihydrofolate accumulates and can be detected in biological samples. The compound is soluble in water and alkaline solutions.
|
| Toxicity/Toxicokinetics |
Toxicity data for dihydrofolate are limited as it is an endogenous metabolite. Accumulation of dihydrofolate due to DHFR inhibition can lead to folate deficiency and impaired DNA synthesis, resulting in cytotoxicity. This mechanism underlies the therapeutic and toxic effects of antifolate drugs such as methotrexate.
|
| References |
Nutrients. 2018 Jun 20;10(6). pii: E795.
|
| Additional Infomation |
Dihydrofolic is a derivative of folic acid, which is converted to tetrahydrofolate by Dihydrofolic reductase. It interacts with bacteria during cell division and is targeted by various drugs to inhibit nucleic acid synthesis. It is a metabolite of E. coli and mice. It is the conjugate acid of Dihydrofolic (2-). Dihydrofolic is a metabolite found or produced in E. coli (K12 strain, MG1655 strain). Dihydrofolic has been reported in Arabidopsis thaliana, humans, and common bean, with relevant data available. Dihydrofolic is a metabolite found or produced in Saccharomyces cerevisiae.
Dihydrofolate is a folic acid derivative that is acted upon by dihydrofolate reductase to produce tetrahydrofolate. Because animals and plants require tetrahydrofolate to synthesize purines and pyrimidines, many drugs target dihydrofolate reductase to prevent nucleic acid synthesis. Dihydrofolate is also known as 7,8-dihydrofolic acid. It is used primarily for research purposes in folate metabolism and drug discovery. |
| Molecular Formula |
C19H21N7O6
|
|---|---|
| Molecular Weight |
443.42
|
| Exact Mass |
443.155
|
| CAS # |
4033-27-6
|
| PubChem CID |
135398604
|
| Appearance |
Typically exists as solid at room temperature
|
| Density |
1.69g/cm3
|
| Index of Refraction |
1.762
|
| LogP |
0.628
|
| Hydrogen Bond Donor Count |
7
|
| Hydrogen Bond Acceptor Count |
10
|
| Rotatable Bond Count |
9
|
| Heavy Atom Count |
32
|
| Complexity |
889
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
O=C(O)CC[C@@H](C(O)=O)NC(C1=CC=C(NCC2=NC3=C(N=C(N)NC3=O)NC2)C=C1)=O
|
| InChi Key |
OZRNSSUDZOLUSN-LBPRGKRZSA-N
|
| InChi Code |
InChI=1S/C19H21N7O6/c20-19-25-15-14(17(30)26-19)23-11(8-22-15)7-21-10-3-1-9(2-4-10)16(29)24-12(18(31)32)5-6-13(27)28/h1-4,12,21H,5-8H2,(H,24,29)(H,27,28)(H,31,32)(H4,20,22,25,26,30)/t12-/m0/s1
|
| Chemical Name |
(2S)-2-[[4-[(2-amino-4-oxo-7,8-dihydro-3H-pteridin-6-yl)methylamino]benzoyl]amino]pentanedioic acid
|
| Synonyms |
NSC-165989 NSC 165989 Dihydrofolate
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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 | 2.2552 mL | 11.2760 mL | 22.5520 mL | |
| 5 mM | 0.4510 mL | 2.2552 mL | 4.5104 mL | |
| 10 mM | 0.2255 mL | 1.1276 mL | 2.2552 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
