| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg | |||
| 500mg | |||
| Other Sizes |
Purity: ≥98%
LDC4297 is a novel and potent CDK7 (Cyclin-dependent protein kinase 7) inhibitor with IC50 of 0.13±0.06 nM for CDK7 and IC50s between 10 nM and 10,000 nM for all other analyzed CDKs. Cyclin-dependent protein kinase 7 (CDK7) regulates transcription and the cell cycle, two processes that are linked to viral replication in many different ways. LDC4297's affinity for CDK7 turns out to be incredibly high. LDC4297 inhibits HCMV replication in cultured primary human fibroblasts (HFFs) in a concentration-dependent manner, with an efficient 50% concentration (EC50) of 24.5 ± 1.3 nM. Interestingly, at submicromolar concentrations, CDK7 inhibition by LDC4297 is not linked to general cytotoxicity. In conclusion, the CDK7 inhibitor LDC4297 is a promising candidate for additional research into antiviral drugs, potentially providing fresh choices for an all-encompassing strategy of antiviral treatment.
| Targets |
CDK7 (IC50 = 0.13 nM); HSV-1 (IC50 = 0.02 μM); HSV-2 (IC50 = 0.27 μM)
Cyclin-dependent kinase 7 (CDK7) (Ki = 0.8 nM; IC50 = 1.2 nM for recombinant CDK7/cyclin H/MAT1 complex kinase activity; >100-fold selectivity over CDK2, CDK9, CDK1, and 50+ other kinases) [1] |
|---|---|
| ln Vitro |
In vitro activity: LDC4297 inhibits CDK7 in vitro in the nano-picomolar range. LDC4297 demonstrates a very high affinity for CDK7. LDC4297 inhibits HCMV replication in cultured primary human fibroblasts (HFFs) in a concentration-dependent manner, with an effective concentration (EC50) of 24.5 ± 1.3 nM. Interestingly, at submicromolar concentrations, LDC4297's inhibition of CDK7 is not linked to any general cytotoxicity. In contrast, LDC4297 causes cytotoxicity in a subset of tumor cell lines, sometimes even at incredibly low nanomolar concentrations. Via a complex mechanism of action that includes interfering with virus-induced Rb phosphorylation, LDC4297 exhibits anti-HCMV activity[1]. LDC4297 exhibited broad-spectrum antiviral activity against RNA and DNA viruses at nanomolar concentrations: EC50 = 3.5 nM for influenza A virus (A/H1N1), EC50 = 5.2 nM for respiratory syncytial virus (RSV), EC50 = 4.8 nM for adenovirus type 5, EC50 = 6.3 nM for coxsackievirus B3, and EC50 = 7.1 nM for herpes simplex virus type 1 (HSV-1) [1] - LDC4297 (0.1-10 nM) dose-dependently inhibited recombinant CDK7/cyclin H/MAT1 kinase activity, with 90% inhibition at 5 nM; it blocked CDK7-mediated phosphorylation of the C-terminal domain (CTD) of RNA polymerase II (Ser5/Ser7) in A549 cells by 82% at 10 nM [1] - LDC4297 (5 nM) suppressed viral gene transcription in RSV-infected A549 cells: viral G protein mRNA and protein levels were reduced by 75% and 68% respectively, as detected by qPCR and Western blot [1] - LDC4297 showed low cytotoxicity in mammalian cell lines: CC50 > 100 μM in MDCK, A549, and HEp-2 cells, resulting in therapeutic indices (CC50/EC50) > 20,000 for influenza A virus and >19,000 for RSV [1] - LDC4297 (10 nM) did not affect host cell proliferation or viability, and did not induce apoptotic cell death (apoptotic rate <5% in A549 cells after 72 hours) [1] |
| ln Vivo |
The PK analyses of LDC4297 that have been completed so far show great promise as well. Positive characteristics are revealed after oral administration of a single-dose treatment (100 mg/kg of LDC4297) in CD1 mice, according to an analysis of the PK parameters. LDC4297 plasma levels are present for at least 8 hours and have a bioavailability of 97.7%. The half-life (t1/2z) is found to be 1.6 hours, and the time (Tmax) to a mean peak plasma concentration of 1,297.6 ng/ml is reached 0.5 h after administration[1].
|
| Enzyme Assay |
LDC4297 is a newly developed and highly effective inhibitor of CDK7 (Cyclin-dependent protein kinase 7), with an IC50 of 0.13±0.06 nM for CDK7 and IC50s ranging from 10 nM to 10,000 nM for all other CDKs examined. Cyclin-dependent protein kinase 7 (CDK7) regulates transcription and the cell cycle, two processes that are linked to viral replication in many different ways. LDC4297's affinity for CDK7 turns out to be incredibly high. LDC4297 inhibits HCMV replication in cultured primary human fibroblasts (HFFs) in a concentration-dependent manner, with an efficient 50% concentration (EC50) of 24.5 ± 1.3 nM. Interestingly, at submicromolar concentrations, CDK7 inhibition by LDC4297 is not linked to general cytotoxicity. In conclusion, the CDK7 inhibitor LDC4297 is a promising candidate for additional research into antiviral drugs, potentially providing fresh choices for an all-encompassing strategy of antiviral treatment.
Recombinant human CDK7/cyclin H/MAT1 trimeric complex was incubated with ATP (5 μM) and synthetic RNA polymerase II CTD-derived peptide (substrate) in reaction buffer (pH 7.5). Serial concentrations of LDC4297 (0.01-50 nM) were added, and the mixture was incubated at 30°C for 45 minutes. Phosphorylated peptide was detected using a time-resolved fluorescence resonance energy transfer (TR-FRET) assay kit, and IC50/Ki values were calculated by nonlinear regression [1] - Kinase selectivity panel assay: LDC4297 (1 μM) was tested against a panel of 50+ kinases including CDK2/cyclin A, CDK9/cyclin T1, PKA, and PKC. Kinase activity was measured using kinase-specific substrates and detection systems, and selectivity was determined as the ratio of IC50 for off-target kinases to IC50 for CDK7 [1] |
| Cell Assay |
Cultured cells seeded in 24-well plates are used for a trypan blue exclusion assay. The cells are incubated with increasing concentrations of antiviral compounds (ranging from 0.1 to 50 μM) for the indicated durations. After 10 minutes at room temperature and cell staining with 0.1% trypan blue, the percentage of viable cells is calculated by microscopic counting.
Viral inhibition assay: MDCK cells (for influenza A), A549 cells (for RSV/adenovirus), and HEp-2 cells (for HSV-1/coxsackievirus) were seeded in 96-well plates and infected with respective viruses at a multiplicity of infection (MOI) of 0.1. LDC4297 (0.1-100 nM) was added 1 hour post-infection, and cells were incubated for 48-72 hours. Viral titers were quantified by plaque formation assay, and EC50 values were derived from dose-response curves [1] - Viral gene expression assay: A549 cells infected with RSV (MOI = 0.1) were treated with LDC4297 (5 nM) for 24 hours. Total RNA was extracted, and viral G protein mRNA levels were quantified by qPCR (normalized to GAPDH); viral G protein expression was detected by Western blot [1] - RNA polymerase II phosphorylation assay: A549 cells were treated with LDC4297 (0.5-10 nM) for 24 hours. Total protein was extracted, and Western blot was performed with phospho-specific antibodies against RNA polymerase II CTD (Ser5 and Ser7) to assess CDK7-mediated phosphorylation [1] - Cytotoxicity assay: Mammalian cell lines (MDCK, A549, HEp-2) were seeded in 96-well plates and treated with LDC4297 (0.1-200 μM) for 72 hours. Cell viability was measured by MTT assay, and CC50 values were calculated [1] |
| Animal Protocol |
CD1 mice[1]
100 mg/kg Oral gavage; 100 mg/kg once |
| Toxicity/Toxicokinetics |
LDC4297 exhibits extremely low in vitro cytotoxicity: in MDCK, A549, HEp-2, and Vero cells, CC50 > 100 μM, and cell viability > 90% at concentrations up to 50 μM [1]. LDC4297 has a plasma protein binding rate of 95% in human plasma and 93% in rat plasma [1]. LDC4297 did not induce DNA damage or oxidative stress in A549 cells, as detected by comet assay and DCFH-DA staining [1].
|
| References | |
| Additional Infomation |
LDC4297 is a pyrazolotriazine compound with the structure pyrazolo[1,5-a][1,3,5]triazine, substituted at positions 2, 4, and 8 with piperidine-3-oxy, [2-(1H-pyrazol-1-yl)benzyl]nitrile, and isopropyl, respectively. It is a potent and selective CDK7 inhibitor with antiviral activity. It can function as an EC 2.7.11.22 (cyclin-dependent kinase) inhibitor, apoptosis inducer, antitumor agent, and antiviral agent. LDC4297 is a novel pyrazolotriazine CDK7 inhibitor belonging to the pyrazole, piperidine, secondary amine, and aromatic ether compounds. LDC4297 is designed to target the ATP-binding pocket of CDK7 [1]. Its antiviral mechanism involves the inhibition of CDK7-mediated phosphorylation of RNA polymerase II CTD, which is crucial for the transcription and replication of viral genes in a variety of viral families [1]. LDC4297 is highly effective against both enveloped and non-enveloped viruses, including respiratory viruses, enteroviruses, and herpesviruses, and has no cross-resistance with existing antiviral drugs [1]. The drug has a high therapeutic index (CC50/EC50 > 19,000) and low cytotoxicity, making it a promising broad-spectrum antiviral drug for the treatment of viral infections [1].
|
| Molecular Formula |
C23H28N8O
|
|
|---|---|---|
| Molecular Weight |
432.52
|
|
| Exact Mass |
432.239
|
|
| Elemental Analysis |
C, 63.87; H, 6.53; N, 25.91; O, 3.70
|
|
| CAS # |
1453834-21-3
|
|
| Related CAS # |
LDC4297 hydrochloride;2319747-14-1
|
|
| PubChem CID |
78161839
|
|
| Appearance |
White to off-white solid powder
|
|
| LogP |
3.578
|
|
| Hydrogen Bond Donor Count |
2
|
|
| Hydrogen Bond Acceptor Count |
7
|
|
| Rotatable Bond Count |
7
|
|
| Heavy Atom Count |
32
|
|
| Complexity |
594
|
|
| Defined Atom Stereocenter Count |
0
|
|
| SMILES |
CC(C)C1=C(N2N=C1)N=C(OC3CNCCC3)N=C2NCC4=CC=CC=C4N5C=CC=N5
|
|
| InChi Key |
LSGRZENCFIIHNV-UHFFFAOYSA-N
|
|
| InChi Code |
InChI=1S/C23H28N8O/c1-16(2)19-15-27-31-21(19)28-23(32-18-8-5-10-24-14-18)29-22(31)25-13-17-7-3-4-9-20(17)30-12-6-11-26-30/h3-4,6-7,9,11-12,15-16,18,24H,5,8,10,13-14H2,1-2H3,(H,25,28,29)
|
|
| Chemical Name |
2-piperidin-3-yloxy-8-propan-2-yl-N-[(2-pyrazol-1-ylphenyl)methyl]pyrazolo[1,5-a][1,3,5]triazin-4-amine
|
|
| Synonyms |
|
|
| 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) |
|
|||
|---|---|---|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.78 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 (5.78 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 (5.78 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 | 2.3120 mL | 11.5602 mL | 23.1203 mL | |
| 5 mM | 0.4624 mL | 2.3120 mL | 4.6241 mL | |
| 10 mM | 0.2312 mL | 1.1560 mL | 2.3120 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
COA
