PROTAC CDK2/4/6 Degrader-2
Based on 1 Customer Validation
PROTAC CDK2/4/6 Degrader-2 is a CDK2/4/6 PROTAC degrader. PROTAC CDK2/4/6 Degrader-2. PROTAC CDK2/4/6 Degrader-2 can be converted into the prodrug PROTAC CDK2/4/6 Degrader-1 (HY-171826) through a one-step reaction with chloromethyl pivalate. PROTAC CDK2/4/6 Degrader-2 degrades CDK2/4/6 and their complex in malignant melanomas cells. PROTAC CDK2/4/6 Degrader-2 induces cell cycle arrest and cell apoptosis in various cancer cells, in particular for melanomas. PROTAC CDK2/4/6 Degrader-2 can be used for malignant melanomas research.
(Pink: CDK2 and CDK4 and CDK6 ligand (HY-168440); Blue: Cereblon ligand (HY-10984); Black: linker).
For research use only. We do not sell to patients.
- Purity : 97.22%
- CAS No.: 2541626-49-5
- Formula: C40H45N9O6
- Molecular Weight:747.84
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Storage:Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 6 months , -20°C, 1 month
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Protocols
- Purity & Documentation
- References
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Help & FAQs
Biological Activity
Description
IC50 & Target
[1]|
CDK2 |
CDK3 |
CDK6 |
Caspase 3 |
CDK4 |
Cellular Effect
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Cell Line
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Type | Value | Description | References |
|---|---|---|---|---|
| A-375 | IC50 |
0.1659 μM
Compound: 3
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Cytotoxicity against human A375 cells assessed as reduction in cell viability after 72 hrs by MTT assay
Cytotoxicity against human A375 cells assessed as reduction in cell viability after 72 hrs by MTT assay
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[PMID: 33256948] |
In Vitro
PROTAC CDK2/4/6 Degrader-2 (compound 3) (72 h) exhibits prominent anti-proliferative activity against B16F10 and A375, with IC50s of 0.09861 and 0.1659 μM, respectively[1].
PROTAC CDK2/4/6 Degrader-2 (0-2000 nM, 0-12 h) mediates CDK2/4/6 protein degradation and impairs downstream Rb signaling activation by hijacking the E3 ubiquitin ligase CRBN in B16F10 and A375 cells[1].
PROTAC CDK2/4/6 Degrader-2 (50-500 nM, 7 days) exerts significant anti-proliferative activity in B16F10 and A375 cells[1].
PROTAC CDK2/4/6 Degrader-2 (125-1000 nM,48 h) significantly induces apoptosis of A375 and B16F10 cells in a dose-dependent manner in B16F10 and A375 cells[1].
PROTAC CDK2/4/6 Degrader-2 (0-2000 nM, 48 h) induces caspase-dependent apoptosis via the common P53/Bcl-2/Bax apoptotic pathway in B16F10 and A375 cells[1].
PROTAC CDK2/4/6 Degrader-2 (25-1000 nM ,48 h) induces cell cycle arrest via the degradation of cell cycle-related proteins CDK2/4/6 in A375 and B16F10 cells[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:B16F10 and A375 cells
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Concentration:0, 62.5, 100, 125, 250, 500, 1000 and 2000 nM
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Incubation Time:0, 3, 6, 9 and 12 h
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Result:Induced significant intracellular cleavage of all CDK 2, CDK 4 and CDK 6 proteins in both concentration-dependent and time dependent manners.
Mitigated downstream p-Rb signaling pathway activation of melanoma cells.
Induced CDK2/4/6 protein degradation via the proteasomal pathway at 100 nM and 6 h.
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Cell Line:B16F10 and A375
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Concentration:50, 100, 250, 500 nM
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Incubation Time:7 days
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Result:Exerted a remarkable inhibiting effect on colony formation in A375 and B16F10 in low nano molar concentration.
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Cell Line:B16F10 and A375 cells
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Concentration:125, 250, 500, 1000 nM
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Incubation Time:48 h
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Result:Significantly induced apoptosis.
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Cell Line:B16F10 and A375 cells
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Concentration:0, 62.5, 125, 250, 500, 1000 and 2000 nM
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Incubation Time:48 h
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Result:Upregulated casepase-3 and cleaved PARP protein expression.
Also significantly induced up-regulation of the pro-protein level of P-53 and Bax.
Downregulated the anti-apoptotic protein level of Bcl-2.
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Cell Line:B16F10 and A375 cells
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Concentration:0, 25, 50, 100, 250, 500, 1000 nM
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Incubation Time:48 h
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Result:Increased accumulation of A375 cell population in G0/G1 phase, but G2/M phase for B16F10 cell.
Parmacokinetics
Chemical Information
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CAS No. 2541626-49-5
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Appearance Solid
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Molecular Weight 747.84
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Formula C40H45N9O6
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Color White to yellow
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SMILES
O=C(C1=CC=C(NC2=NC3=C(C=C(N3C4CCCC4)C(N(C)C)=O)C=N2)C=C1)NCCCCCCNC5=CC=CC(C(N6C7C(NC(CC7)=O)=O)=O)=C5C6=O
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Solvent & Solubility
In Vitro:
DMSO : 200 mg/mL (267.44 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
In Vivo:
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 5 mg/mL (6.69 mM); Suspended solution
This protocol yields a suspended solution of ≥ 5 mg/mL (saturation unknown). Suspended solution can be used for oral and intraperitoneal injection.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (50.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL.
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Protocols
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Apoptosis
Apoptosis, also called programmed cell death, is generally characterized by distinct morphological characteristics.
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TUNEL staining for apoptotic DNA fragmentation
TUNEL staining detects DNA strand breaks by using terminal deoxynucleotidyl transferase to add labeled nucleotides to exposed 3′-OH DNA termini, generating either microscopic staining in fixed cells or tissue sections, or fluorescence/cytometric signal in cell suspensions. TUNEL positivity reflects DNA fragmentation but should not be interpreted alone as definitive apoptosis, because TUNEL can also label necrotic, autolytic, mechanically damaged, or DNA-repair-associated DNA breaks.
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Flow cytometric DNA-content cell-cycle staining
Flow cytometric DNA-content cell-cycle staining measures the fluorescence intensity of DNA-bound fluorochromes in single cells or nuclei to estimate DNA content distributions, allowing assignment of populations to G0/G1, S, and G2/M phases by DNA histogram deconvolution. Propidium iodide (PI) intercalates into DNA, and PI fluorescence is proportional to cellular DNA content when staining is performed under conditions that make DNA accessible and minimize non-DNA signal. Cells with G2/M DNA content are expected to show approximately twice the fluorescence intensity of G0/G1 cells, while S-phase cells occupy intermediate fluorescence values. PI-based DNA-content analysis can also detect cells with fractional DNA content, often reported as sub-G1, when DNA fragmentation and extraction during staining reduce retained DNA signal in apoptotic cells. DAPI is an alternative DNA fluorochrome for univariate DNA-content analysis, while bivariate approaches combining DNA content with proliferation
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Annexin V plus membrane-impermeant dye apoptosis staining
Annexin V-based apoptosis assays rely on the detection of phosphatidylserine (PS) externalization from the inner leaflet of the plasma membrane to the outer leaflet, an early biochemical hallmark of apoptosis. Fluorescently labeled Annexin V binds PS in a calcium-dependent manner, enabling identification of early apoptotic cells by flow cytometry or fluorescence microscopy. When combined with a membrane-impermeant DNA-binding dye (e. g. , propidium iodide), this approach allows discrimination between viable (Annexin V−/dye−), early apoptotic (Annexin V+/dye−), and late apoptotic or necrotic (Annexin V+/dye+) cell populations by assessing membrane integrity and PS exposure.
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BrdU Incorporation Assay
Bromodeoxyuridine (BrdU) incorporation assay is based on the principle that BrdU, a thymidine analog, is incorporated into newly synthesized DNA during the S phase of the cell cycle, thereby serving as a marker of DNA replication and cellular proliferation. Incorporated BrdU can be detected using anti-BrdU antibodies following DNA denaturation, enabling visualization or quantification of proliferating cells through immunochemical detection methods such as immunofluorescence or immunohistochemistry.
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Apoptosis Solutions
Apoptosis is a regulated, generally non-lytic cell-death pathway that removes unwanted, damaged, infected, or abnormal cells through coordinated morphological changes, caspase activation, DNA fragmentation, and membrane remodeling. The intrinsic apoptosis pathway is controlled mainly by mitochondrial outer membrane permeabilization, BCL-2 family proteins, cytochrome c release, apoptosome formation, caspase-9 activation, and downstream executioner caspase-3/7 activation. The extrinsic apoptosis pathway is initiated by death receptors such as Fas, TNFR, and TRAIL receptors, which recruit adaptor proteins and activate caspase-8 before engaging executioner caspases or mitochondrial amplification through BID cleavage. Apoptosis is linked to many phenotypes, including cancer cell killing, tissue homeostasis, immune regulation, neurodegeneration, infection response, and treatment-induced cytotoxicity; unresolved questions include how apoptosis interacts with necroptosis, pyroptosis, ferroptos
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Protocol for Cell Cycle
Cell-cycle analysis by flow cytometry measures DNA content in single cells to estimate the fraction of cells in G0/G1, S, and G2/M phases. Propidium iodide intercalates into DNA, and after RNA removal with RNase, fluorescence intensity reflects cellular DNA content: 2N cells are assigned to G0/G1, cells between 2N and 4N to S phase, and 4N cells to G2/M. DNA-content analysis alone cannot reliably separate G0 from G1 or G2 from M. Ki-67 can distinguish quiescent G0 cells from cycling cells, EdU or BrdU incorporation marks active DNA synthesis in S phase, and phospho-histone H3 staining identifies mitotic cells within the 4N population.
Purity & Documentation
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Data Sheet (275 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Handling Instructions (2659 KB)
References
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 1.3372 mL | 6.6859 mL | 13.3718 mL | 33.4296 mL |
| 5 mM | 0.2674 mL | 1.3372 mL | 2.6744 mL | 6.6859 mL | |
| 10 mM | 0.1337 mL | 0.6686 mL | 1.3372 mL | 3.3430 mL | |
| 15 mM | 0.0891 mL | 0.4457 mL | 0.8915 mL | 2.2286 mL | |
| 20 mM | 0.0669 mL | 0.3343 mL | 0.6686 mL | 1.6715 mL | |
| 25 mM | 0.0535 mL | 0.2674 mL | 0.5349 mL | 1.3372 mL | |
| 30 mM | 0.0446 mL | 0.2229 mL | 0.4457 mL | 1.1143 mL | |
| 40 mM | 0.0334 mL | 0.1671 mL | 0.3343 mL | 0.8357 mL | |
| 50 mM | 0.0267 mL | 0.1337 mL | 0.2674 mL | 0.6686 mL | |
| 60 mM | 0.0223 mL | 0.1114 mL | 0.2229 mL | 0.5572 mL | |
| 80 mM | 0.0167 mL | 0.0836 mL | 0.1671 mL | 0.4179 mL | |
| 100 mM | 0.0134 mL | 0.0669 mL | 0.1337 mL | 0.3343 mL |
PROTAC CDK2/4/6 Degrader-2 Related Classifications
HY-185121 Related Classifications
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