| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
Ki: 5.1 nM (WT), 34.3 nM (L28R)[1]
4′-DTMP targets dihydrofolate reductase (DHFR), a key enzyme in the folate biosynthesis pathway that catalyzes the reduction of dihydrofolate to tetrahydrofolate. The compound is a potent DHFR inhibitor with Ki values of 5.1 nM for wild-type DHFR (DHFRWT) and 34.3 nM for the L28R variant (DHFRL28R). 4′-DTMP carries a polar modification that induces additional local interactions with the enzyme. The compound's ability to inhibit both wild-type and resistant DHFR variants makes it a valuable tool for studying antibiotic resistance mechanisms and developing improved antimicrobial agents. |
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| ln Vitro |
4′-DTMP demonstrates potent DHFR inhibitory activity in vitro. It has Ki values of 5.1 nM for wild-type DHFR and 34.3 nM for the DHFRL28R variant. The compound's polar modification induces additional local interactions with the enzyme, enhancing its binding affinity. 4′-DTMP exhibits antimicrobial activity against Escherichia coli. Studies have shown that 4′-DTMP selects against the emergence of TMP-resistant bacteria carrying the L28R mutation in laboratory experiments. These in vitro findings demonstrate the compound's potential as a research tool for studying DHFR inhibition and bacterial resistance mechanisms.
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| ln Vivo |
In vivo activity data for 4′-DTMP are not extensively documented in the available literature. The compound is primarily used as a research tool for in vitro biochemical and cellular studies of DHFR inhibition. Its antimicrobial activity has been demonstrated against Escherichia coli in vitro, and it has been shown to select against the emergence of TMP-resistant bacteria in laboratory evolution experiments. Further in vivo studies, including efficacy in animal models of bacterial infection, would be required to fully characterize the compound's therapeutic potential.
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| Enzyme Assay |
The in vitro enzyme assay for 4′-DTMP involves measuring its inhibition of DHFR enzymatic activity. Recombinant wild-type DHFR and the L28R variant are expressed and purified. Enzyme activity is assessed by spectrophotometrically monitoring the reduction of dihydrofolate to tetrahydrofolate, coupled to the oxidation of NADPH. 4′-DTMP is incubated with the enzyme and substrate at various concentrations. Inhibition constants (Ki) are determined by analyzing the enzyme kinetics using Lineweaver-Burk or Dixon plots. The assay buffer typically contains Tris-HCl, KCl, and DTT to maintain enzyme stability and activity.
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| Cell Assay |
In vitro cellular assays for 4′-DTMP typically use bacterial cultures, particularly Escherichia coli, to assess antimicrobial activity. Minimal inhibitory concentration (MIC) assays are performed by culturing bacteria in liquid medium with serial dilutions of the compound. Bacterial growth is monitored by measuring optical density at 600 nm. The compound's ability to inhibit bacterial growth is quantified as the MIC. Resistance evolution experiments are conducted by serial passaging bacteria in the presence of sub-inhibitory concentrations of 4′-DTMP to assess its ability to select against resistance emergence.
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| Animal Protocol |
In vivo animal experiments for 4′-DTMP are not extensively documented. As a research compound primarily used for in vitro studies, 4′-DTMP has not been widely tested in animal models. Standard in vivo efficacy studies would involve mouse models of bacterial infection, where the compound would be administered via oral or parenteral routes. Bacterial load in tissues would be measured to assess antimicrobial efficacy. Pharmacodynamic studies could evaluate the compound's effects on bacterial growth and resistance development in vivo.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of 4′-DTMP are not extensively documented. As a small-molecule DHFR inhibitor with a molecular weight of 276.29, the compound is expected to have reasonable bioavailability. Its polar modification may influence its absorption and tissue distribution. 4′-DTMP is supplied as a solid and as a DMSO solution for research use. Further pharmacokinetic studies, including assessments of absorption, distribution, metabolism, and excretion (ADME), are necessary to fully characterize its PK profile and support its development as a therapeutic agent.
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| Toxicity/Toxicokinetics |
Toxicological data for 4′-DTMP are not extensively available in the public domain. As a research-grade compound intended for in vitro biochemical and cellular studies, 4′-DTMP has not undergone extensive toxicological evaluation. Standard cytotoxicity assays in mammalian cell lines may have been performed to assess safety margins. The compound's antimicrobial activity suggests selectivity for bacterial DHFR over mammalian DHFR, which may contribute to a favorable toxicity profile. Further preclinical toxicology studies would be required before clinical development.
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| References | |
| Additional Infomation |
4′-DTMP (4-Demethyltrimethoprim) is a potent DHFR inhibitor with Ki values of 5.1 nM for wild-type DHFR and 34.3 nM for the L28R variant. It carries a polar modification that induces additional local interactions with the enzyme. The compound has antimicrobial activity against Escherichia coli and selects against the emergence of TMP-resistant bacteria. 4′-DTMP is a research tool for studying DHFR inhibition, antibiotic resistance, and bacterial evolution. It has a molecular weight of 276.29 and formula C13H16N4O3. No clinical trials or regulatory approvals have been reported.
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| Molecular Formula |
C13H16N4O3
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|---|---|
| Molecular Weight |
276.29
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| Exact Mass |
276.122
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| CAS # |
21253-58-7
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| PubChem CID |
10423570
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| Appearance |
Brown to dark brown solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
567.4±60.0 °C at 760 mmHg
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| Flash Point |
296.9±32.9 °C
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| Vapour Pressure |
0.0±1.6 mmHg at 25°C
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| Index of Refraction |
1.652
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| LogP |
-0.09
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
20
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| Complexity |
293
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| Defined Atom Stereocenter Count |
0
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| SMILES |
COC1=CC(=CC(=C1O)OC)CC2=CN=C(N=C2N)N
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| InChi Key |
HPOCGNHBIFZCAN-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C13H16N4O3/c1-19-9-4-7(5-10(20-2)11(9)18)3-8-6-16-13(15)17-12(8)14/h4-6,18H,3H2,1-2H3,(H4,14,15,16,17)
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| Chemical Name |
4-[(2,4-diaminopyrimidin-5-yl)methyl]-2,6-dimethoxyphenol
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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) |
DMSO: 50 mg/mL (180.97 mM)
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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 | 3.6194 mL | 18.0969 mL | 36.1939 mL | |
| 5 mM | 0.7239 mL | 3.6194 mL | 7.2388 mL | |
| 10 mM | 0.3619 mL | 1.8097 mL | 3.6194 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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