| Size | Price | Stock | Qty |
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| 250mg |
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| 500mg |
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| Other Sizes |
| Targets |
DHFR-IN-3 targets dihydrofolate reductase (DHFR), an enzyme that catalyzes the reduction of dihydrofolate to tetrahydrofolate. This reaction is crucial for the synthesis of purines, thymidylate, and several amino acids, making DHFR a vital target for antimicrobial and anticancer therapies. By inhibiting DHFR, the compound depletes tetrahydrofolate pools, thereby disrupting DNA synthesis and cell division. DHFR-IN-3 has shown inhibitory activity against DHFR from different species, including rat liver and *Pneumocystis carinii*, with IC50 values of 19 µM and 12 µM, respectively. This suggests a degree of species selectivity, which is of interest for developing selective inhibitors.
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| ln Vitro |
In vitro studies have demonstrated that DHFR-IN-3 is a potent inhibitor of DHFR enzymatic activity. Its inhibitory potency has been quantified in cell-free assays, showing IC50 values of 19 µM against rat liver DHFR and 12 µM against *P. carinii* DHFR. The compound's activity is attributed to its ability to bind to the active site of the DHFR enzyme, competing with the natural substrate dihydrofolate. This inhibition is dose-dependent, and the compound has been used to study the structure-activity relationships of diaminoquinazolines as DHFR inhibitors. It is classified as an antifolate agent and serves as a valuable tool for investigating the role of DHFR in various cellular processes.
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| ln Vivo |
In vivo activity for DHFR-IN-3 has not been extensively documented in the available literature. As a research chemical, its primary use is for in vitro enzyme inhibition studies and biochemical assays. While DHFR inhibitors are known to have therapeutic potential, specific in vivo efficacy and pharmacological data for this particular compound are limited. However, the compound's structural class and mechanism of action suggest it could be explored for in vivo applications, particularly in models of infections or cancers where DHFR is a validated target. Further research is needed to characterize its pharmacokinetic and pharmacodynamic properties in living organisms.
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| Enzyme Assay |
In vitro enzyme assays for DHFR-IN-3 involve measuring the inhibition of DHFR-catalyzed reduction of dihydrofolate to tetrahydrofolate. A typical protocol uses recombinant or tissue-derived DHFR (e.g., from rat liver or *P. carinii*) incubated with the substrate dihydrofolate and the cofactor NADPH in a buffer solution. Varying concentrations of DHFR-IN-3 are added to the reaction mixture, and the enzymatic activity is monitored spectrophotometrically by measuring the decrease in absorbance at 340 nm, which corresponds to the oxidation of NADPH. The IC50 values are calculated by plotting the percentage of enzyme inhibition against the compound's concentration. The compound is typically dissolved in DMSO for stock solutions and diluted in the assay buffer.
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| Cell Assay |
In vitro cell-based assays for DHFR-IN-3 are typically performed in cell lines that are dependent on folate metabolism for proliferation. Cells are seeded in multi-well plates and treated with varying concentrations of the compound for a specified duration, typically 48-72 hours. Cell viability is assessed using standard assays such as MTT, CCK-8, or by measuring DNA synthesis through the incorporation of radioactive or fluorescent nucleotide analogs. The compound's anti-proliferative effect is determined by calculating the half-maximal inhibitory concentration (IC50) from dose-response curves. For mechanistic studies, the effects of DHFR-IN-3 on cell cycle progression and apoptosis can be evaluated using flow cytometry. The compound is typically dissolved in DMSO and diluted in cell culture medium to the desired concentrations.
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| Animal Protocol |
Specific in vivo animal experiment protocols for DHFR-IN-3 are not widely reported. However, for in vivo studies, DHFR-IN-3 can be formulated for administration. A recommended formulation for similar compounds involves dissolving the compound in a mixture of solvents, such as 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline, to achieve a clear solution at a concentration of ≥ 2.5 mg/mL. Another formulation option is 10% DMSO in 90% corn oil. The route of administration (e.g., oral, intraperitoneal, or intravenous) and dosing regimen would depend on the specific experimental design and disease model being studied.
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| ADME/Pharmacokinetics |
Pharmacokinetic (PK) properties of DHFR-IN-3 have not been extensively characterized in the available literature. However, some physicochemical properties relevant to its PK can be inferred. The compound has a molecular weight of 239.07 g/mol and is soluble in DMSO. For in vivo formulation, it can be prepared in a mixture of solvents such as 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline, achieving a clear solution at ≥ 2.5 mg/mL. The compound is recommended to be stored as a powder at 4°C, protected from light, and in solution at -80°C for up to 6 months or at -20°C for up to 1 month.
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| Toxicity/Toxicokinetics |
Specific toxicological data for DHFR-IN-3 are not provided in the available sources. As a research chemical, its safety profile in humans has not been established. The compound is classified as a research reagent for in vitro and biochemical studies and is not for therapeutic or veterinary use. Standard laboratory safety precautions should be followed when handling DHFR-IN-3, including the use of appropriate personal protective equipment. Its purity is typically ≥95% or higher.
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| References | |
| Additional Infomation |
DHFR-IN-3 is a small-molecule inhibitor of dihydrofolate reductase (DHFR), identified as 7-bromoquinazoline-2,4-diamine. It is a quinazoline derivative that acts as an antifolate agent. The compound has been studied for its inhibitory activity against DHFR from different species, showing IC50 values of 19 µM and 12 µM against rat liver and *P. carinii* DHFR, respectively. It is used as a research tool to investigate the role of DHFR in cell proliferation and as a potential lead compound for developing antimicrobial or anticancer agents. The compound is soluble in DMSO and is stored at 4°C, protected from light. DHFR-IN-3 is intended for research use only and is not for human consumption.
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| Molecular Formula |
C8H7BRN4
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| Molecular Weight |
239.0720
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| Exact Mass |
237.985
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| CAS # |
137553-43-6
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| PubChem CID |
456247
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| Appearance |
White to off-white solid powder
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| Density |
1.8±0.1 g/cm3
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| Boiling Point |
509.4±58.0 °C at 760 mmHg
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| Melting Point |
255-256°C
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| Flash Point |
261.9±32.3 °C
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| Vapour Pressure |
0.0±1.3 mmHg at 25°C
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| Index of Refraction |
1.811
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| LogP |
1.73
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
13
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| Complexity |
189
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
YPWBLOIRVSSESG-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C8H7BrN4/c9-4-1-2-5-6(3-4)12-8(11)13-7(5)10/h1-3H,(H4,10,11,12,13)
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| Chemical Name |
7-bromoquinazoline-2,4-diamine
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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 Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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) |
DMSO : ~100 mg/mL (~418.29 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (10.46 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (10.46 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (10.46 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.1829 mL | 20.9144 mL | 41.8288 mL | |
| 5 mM | 0.8366 mL | 4.1829 mL | 8.3658 mL | |
| 10 mM | 0.4183 mL | 2.0914 mL | 4.1829 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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