| Targets |
(S)-JWZ-5-13 targets cyclin-dependent kinase 7 (CDK7), which is a key regulator of the cell cycle and transcription. CDK7 functions as a CDK-activating kinase (CAK) that phosphorylates and activates other CDKs (CDK1, CDK2, CDK4, CDK6) involved in cell cycle progression, and as a component of the transcription factor TFIIH that phosphorylates RNA polymerase II. As a PROTAC negative control, (S)-JWZ-5-13 binds to CDK7 with similar affinity to the active degrader but lacks the ability to recruit an E3 ubiquitin ligase due to its stereochemistry, preventing formation of the ternary complex required for ubiquitination and degradation.
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| ln Vitro |
In vitro, (S)-JWZ-5-13 inhibits CDK7 with an IC50 of 21.1 nM, which is similar to the active PROTAC degrader JWZ-5-13. Despite this comparable enzyme inhibition activity, (S)-JWZ-5-13 fails to induce CDK7 degradation in cells. This dissociation between CDK7 inhibition and degradation demonstrates that the compound binds to CDK7 with high affinity in a cell-free system but does not engage the E3 ligase machinery in the cellular context. The compound therefore serves as an ideal negative control for evaluating PROTAC-mediated CDK7 degradation studies, allowing researchers to attribute CDK7 protein level reduction to targeted degradation rather than kinase inhibition or transcriptional effects.
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| ln Vivo |
No direct in vivo activity studies have been published for (S)-JWZ-5-13. As a negative control compound, it is not expected to have significant in vivo efficacy. The active PROTAC JWZ-5-13 has shown oral bioavailability in mice and effectively degrades CDK7 in multiple cancer cells, leading to potent inhibition of cell proliferation. In contrast, (S)-JWZ-5-13 likely lacks in vivo efficacy due to its inability to induce CDK7 degradation, serving as a control in pharmacokinetic/pharmacodynamic studies to confirm that observed in vivo effects are due to CDK7 degradation rather than kinase inhibition.
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| Enzyme Assay |
No specific enzyme/receptor binding protocols have been established for (S)-JWZ-5-13. For CDK7 kinase inhibition studies, a radiometric protein kinase assay or fluorescence-based kinase activity assay is used. Recombinant CDK7/cyclin H/MAT1 complex is incubated with varying concentrations of (S)-JWZ-5-13 (0.1-1000 nM) in assay buffer containing ATP and a peptide substrate. The reaction is initiated by adding ATP and incubated at 30degC. Incorporated radioactivity (for radiometric assay) or fluorescence signal (for fluorescence-based assay) is measured. IC50 values are calculated from dose-response curves. For binding affinity studies, surface plasmon resonance (SPR) or fluorescence polarization can be used to measure direct binding to CDK7. The same protocols apply for the active degrader.
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| Cell Assay |
In vitro cellular protocols for (S)-JWZ-5-13 involve culturing cancer cell lines such as HeLa, HCT116, or other CDK7-dependent cells in DMEM with 10% FBS at 37degC in 5% CO2. Cells are seeded in 6-well or 12-well plates at 1-2×10⁵ cells/well and allowed to attach overnight. Cells are treated with varying concentrations of (S)-JWZ-5-13 (0.1 nM to 10 microM) for 4-24 hours. For comparison, parallel treatments with the active PROTAC JWZ-5-13 are performed. CDK7 protein levels are assessed by western blotting of cell lysates using anti-CDK7 antibody, with GAPDH or beta-actin as loading control. Unlike the active degrader, (S)-JWZ-5-13 should show no reduction in CDK7 protein levels. Cell viability is assessed using MTT or CellTiter-Glo assays after 48-72 hours. All experiments are performed in triplicate.
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| Animal Protocol |
No in vivo animal protocols have been specifically established for (S)-JWZ-5-13. For active JWZ-5-13, standard xenograft models in immunodeficient mice are used to evaluate CDK7 degradation and anti-tumor efficacy. Approximately 5×10⁶ cancer cells are injected subcutaneously. When tumors reach ~150-200 mm3, mice are treated with JWZ-5-13 orally or intraperitoneally. For (S)-JWZ-5-13 as a control, the same dosing regimen (e.g., 10-50 mg/kg) would be used to demonstrate lack of CDK7 degradation and anti-tumor activity. Tumor volumes are measured every 2-3 days. At study termination, tumors are excised and CDK7 protein levels are analyzed by western blotting to confirm that the active degrader, but not the control compound, reduces CDK7 in vivo. All procedures require approval by institutional animal care committees.
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| ADME/Pharmacokinetics |
No direct pharmacokinetic studies have been published for (S)-JWZ-5-13. The active PROTAC JWZ-5-13 has demonstrated oral bioavailability in mice, indicating favorable absorption properties. (S)-JWZ-5-13 likely has similar physicochemical properties to JWZ-5-13, including molecular weight 983.23 g/mol, predicted logP of approximately 3-4, and solubility in DMSO (~100 mg/mL). For formulation, 10% DMSO, 40% PEG300, 5% Tween 80, 45% saline can be used. Specific PK parameters such as half-life, Cmax, AUC, clearance, and oral bioavailability are reported for JWZ-5-13 but not specifically for the (S)-enantiomer negative control.
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| Toxicity/Toxicokinetics |
According to safety data, (S)-JWZ-5-13 is for research use only and not for human therapeutic applications. Standard laboratory safety precautions should be followed: wear protective gloves, safety goggles, and a lab coat. Work in a well-ventilated fume hood. Avoid inhalation, ingestion, and contact with skin and eyes. The compound should be stored as a powder at -20degC for up to 3 years or at 4degC for 2 years. In solution, store at -80degC for 6 months or -20degC for 1 month. As a PROTAC CDK7 inhibitor, it may have anti-proliferative activity and should be handled with caution. This product is not for human therapeutic use.
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| References | |
| Additional Infomation |
(S)-JWZ-5-13 is a PROTAC CDK7 degrader that serves as a negative control for JWZ-5-13 in PROTAC research. It exhibits similar in vitro CDK7 inhibition (IC50 = 21.1 nM) but fails to induce CDK7 degradation in cells, providing a valuable tool for distinguishing between target binding/degradation and off-target effects. CDK7 is a dual-function kinase that regulates both cell cycle progression (via activation of CDK1/2/4/6) and transcription (via RNA polymerase II phosphorylation), making it an attractive target for cancer therapy, particularly in cancers with MYC or other transcription factor dependencies. The stereochemistry at the linker attachment site prevents E3 ligase recruitment while maintaining target binding affinity. No clinical trial data is available; the compound is for research use only and not approved for human therapy.
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| Related CAS # |
JWZ-5-13
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| Appearance |
White to off-white solid powder
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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 (~101.71 mM; with sonication)
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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.) |
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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