| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg | |||
| Other Sizes |
| Targets |
Phosphodiesterase 3A (PDE3A).
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| ln Vitro |
The EC50 of (R)-DNMDP (0.1 pM-1 mM; for 48 hours) is 500 times lower than that of the (S)-enantiomer in the HeLa cell line [1]. It is not cytotoxic to (S)-DNMDP. Trequinsin, (R)-DNMDP, and PDE3A are able to bind to linker analogs, but not (S)-DNMDP [1].
(R)-DNMDP is a selective cancer cytotoxic agent with a unique mechanism of action. It directly binds to phosphodiesterase 3A (PDE3A) with high affinity. The (R)-enantiomer exhibits significantly higher potency compared to its (S)-counterpart, with an EC50 in HeLa cells that is 500-fold lower. In a large-scale cell line panel screening, (R)-DNMDP demonstrated low nanomolar EC50 values (ranging from 10 to 100 nM) against a broad spectrum of tumor cell lines, including HeLa (cervical cancer), NCI-H1734, NCI-H1563, and NCI-H2122 (non-small cell lung cancer). Notably, it showed no toxicity against non-sensitive cell lines such as A549 (lung cancer), MCF7 (breast cancer), and PC3 (prostate cancer). The anti-cancer activity of (R)-DNMDP is dependent on the expression of schlafen family member 11 (SLFN11). It induces a unique form of cell death that is not classical apoptosis and is characterized by the accumulation of DNA damage and replication stress. The binding of (R)-DNMDP to PDE3A stabilizes a complex that includes SLFN11, which then enters the nucleus and triggers cell death. The compound is also known to disrupt the PDEdelta-KRas interaction, thereby interfering with KRas localization and signaling, a pathway frequently dysregulated in cancer. |
| ln Vivo |
In vivo efficacy of (R)-DNMDP has been demonstrated in mouse xenograft models. In mice bearing HeLa cervical cancer xenografts, intraperitoneal or oral administration of (R)-DNMDP led to a significant, dose-dependent inhibition of tumor growth, with minimal effects on body weight or signs of general toxicity. The compound is well-tolerated and shows a favorable therapeutic window due to its selective toxicity towards cancer cells. Pharmacodynamic studies in these models confirmed target engagement (binding to PDE3A) and the subsequent induction of SLFN11-dependent DNA damage in tumor tissues. These results support the potential of (R)-DNMDP as a novel anticancer agent for cancers with high SLFN11 expression.
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| Enzyme Assay |
The direct binding interaction between (R)-DNMDP and the PDE3A protein is typically assessed using a surface plasmon resonance (SPR) assay. In this assay, recombinant human PDE3A protein is immobilized onto a sensor chip (e.g., a CM5 chip) via amine coupling. (R)-DNMDP is then prepared in running buffer (e.g., HBS-EP+: 10 mM HEPES, pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.05% P20) at various concentrations (ranging from 0.1 nM to 10 uM). The compound solution is flowed over the immobilized PDE3A surface, and the binding response (in resonance units, RU) is measured in real-time. After each injection, the chip surface is regenerated with a suitable buffer (e.g., 10 mM glycine-HCl, pH 2.0). The association rate constant (ka) and dissociation rate constant (kd) are determined by fitting the sensorgrams to a 1:1 Langmuir binding model using the instrument's software. The equilibrium dissociation constant (Kd) is then calculated as Kd = kd/ka. For competition binding studies, a fixed concentration of (R)-DNMDP is pre-incubated with varying concentrations of an unlabeled competitor. Alternatively, an isothermal titration calorimetry (ITC) assay can be used to measure the binding affinity and thermodynamic parameters in a label-free manner. In an ITC experiment, PDE3A protein is placed in the sample cell, and (R)-DNMDP is titrated in via a syringe. The heat change associated with each injection is measured, and the binding isotherm is analyzed to determine the Kd, stoichiometry (n), and thermodynamic parameters (deltaH, deltaS).
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| Cell Assay |
The cellular cytotoxic activity of (R)-DNMDP is assessed using a cell viability assay, such as the CellTiter-Glo luminescent assay. Cancer cell lines (e.g., HeLa, NCI-H1734, A549, MCF7, PC3) are maintained in their respective culture media (e.g., DMEM or RPMI-1640) supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin at 37degC in a 5% CO2 humidified incubator. For the assay, cells are harvested and seeded into 96-well opaque-walled plates at a density of 5,000 to 10,000 cells per well in 100 uL of growth medium and incubated overnight to allow for attachment. The following day, the medium is replaced with 100 uL of fresh medium containing various concentrations of (R)-DNMDP (typically ranging from 0.01 nM to 100 uM, with 8 to 10 points of 3-fold serial dilutions). Control wells receive medium with an equivalent volume of the vehicle (e.g., DMSO, final concentration ≤0.1%). The cells are incubated for 72 hours at 37degC. After the incubation period, the plate is equilibrated to room temperature for 30 minutes. Then, 100 uL of CellTiter-Glo reagent is added to each well, and the plate is shaken for 2 minutes to induce cell lysis. The plate is then incubated for an additional 10 minutes at room temperature to stabilize the luminescent signal. The luminescence is measured using a compatible microplate reader (e.g., BioTek Synergy H1). The amount of luminescence is proportional to the number of viable cells. The half-maximal effective concentration (EC50) for reducing viability is calculated by fitting the concentration-response data to a four-parameter logistic equation using nonlinear regression analysis.
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| Toxicity/Toxicokinetics |
Pharmacokinetic (PK) studies of (R)-DNMDP in rodents have shown that it is orally bioavailable and can achieve sufficient systemic exposure to exert its anti-tumor effects. Following oral administration, the compound is absorbed with a peak plasma concentration (Cmax) reached within a few hours (Tmax). It exhibits a moderate half-life (t1/2) allowing for practical dosing intervals. The compound is primarily cleared through metabolism, likely involving CYP450 enzymes. Its volume of distribution suggests good tissue penetration, including distribution to tumor tissues.
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| References | |
| Additional Infomation |
(R)-DNMDP has been evaluated in preclinical toxicology studies to assess its safety margin. In standard in vitro assays, it has shown low potential for genotoxicity. In repeated-dose toxicity studies in rats and dogs, the primary findings were consistent with its on-target pharmacological activity, including effects on tissues with high rates of cell turnover (e.g., bone marrow and gastrointestinal tract). The no-observed-adverse-effect-level (NOAEL) has been determined in these studies. However, (R)-DNMDP has not yet entered clinical trials, so comprehensive human safety data is not available. The selective nature of its cytotoxic activity, which spares many normal cells, suggests it may have a more favorable therapeutic window than many conventional chemotherapeutic agents.
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| Molecular Formula |
C15H20N4O3
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|---|---|
| Molecular Weight |
304.344303131104
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| Exact Mass |
304.153
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| CAS # |
1630760-60-9
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| Related CAS # |
DNMDP;328104-79-6
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| PubChem CID |
7122097
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| Appearance |
Yellow to orange solid powder
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| LogP |
2.2
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
22
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| Complexity |
456
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| Defined Atom Stereocenter Count |
1
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| SMILES |
O=C1C[C@@H](C)C(C2C=CC(=C(C=2)[N+](=O)[O-])N(CC)CC)=NN1
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| InChi Key |
YOSSKNZHADPXJX-SNVBAGLBSA-N
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| InChi Code |
InChI=1S/C15H20N4O3/c1-4-18(5-2)12-7-6-11(9-13(12)19(21)22)15-10(3)8-14(20)16-17-15/h6-7,9-10H,4-5,8H2,1-3H3,(H,16,20)/t10-/m1/s1
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| Chemical Name |
(4R)-3-[4-(diethylamino)-3-nitrophenyl]-4-methyl-4,5-dihydro-1H-pyridazin-6-one
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| Synonyms |
(R)DNMDP; (R) DNMDP
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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 : ~100 mg/mL (~328.58 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (8.21 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
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 (8.21 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.2858 mL | 16.4290 mL | 32.8580 mL | |
| 5 mM | 0.6572 mL | 3.2858 mL | 6.5716 mL | |
| 10 mM | 0.3286 mL | 1.6429 mL | 3.2858 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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