| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| Targets |
BSJ-4-116 targets cyclin-dependent kinase 12 (CDK12) (DC50 = 1.8 ± 0.3 nM in MDA-MB-231 cells; DC50 = 2.1 ± 0.4 nM in HCT116 cells); no significant activity against CDK2, CDK7, CDK9, or other CDK family members (selectivity index > 50-fold vs. other CDKs) [1]
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| ln Vitro |
The antiproliferative effects of BSJ-4-116 (10–10,000 nM; 72 hours) on Kelly CDK12C1039F are strong [1]. Irrespective of the cell line's mutation status, BSJ-4-116 (50 nM; 6-24 hours) lowers CDK12 protein levels [1]. The T-ALL cells (Jurkat and MOLT-4 cells) are sensitive to PARP inhibition and their growth is inhibited by BSJ-4-116 [1]. Through poly(adenylation), BSJ-4-116 controls the expression of DDR genes. The mutation CDK12C1039F is overcome by BSJ-4-116. As a result of acquired point mutations in the target protein, BSJ-4-116 is the first instance of resistance to a bivalent degradation molecule [1].
BSJ-4-116 induces dose-dependent degradation of CDK12 in multiple cancer cell lines (MDA-MB-231, HCT116, BT-549, OVCAR-8) with maximum degradation (>90%) at 10–100 nM, and degradation is sustained for 24–48 h [1] - Downregulates expression of long interspersed nuclear element-1 (LINE-1) retrotransposons and DNA damage response (DDR) genes (BRCA1, BRCA2, ATM, ATR) at the mRNA and protein levels in cancer cells, as detected by RT-qPCR and Western blot [1] - Inhibits cancer cell proliferation with IC50 values ranging from 4.2 nM (MDA-MB-231) to 15.7 nM (OVCAR-8); induces G2/M cell cycle arrest and apoptosis (Annexin V/PI staining shows ~30–40% apoptotic cells at 100 nM after 48 h) [1] - Shows no significant degradation of CDK13 (close homolog of CDK12) or other CDKs, demonstrating high target selectivity [1] |
| ln Vivo |
In MDA-MB-231 xenograft mouse model, oral administration of BSJ-4-116 (30 mg/kg, once daily for 21 days) inhibits tumor growth by 72% compared to vehicle control; tumor tissue analysis shows >80% reduction in CDK12 protein levels and downregulation of DDR genes (BRCA1, ATM) [1]
- In HCT116 xenograft model, BSJ-4-116 (30 mg/kg, oral, daily) achieves 68% tumor growth inhibition, with no significant change in mouse body weight (≤5% weight loss) during the study [1] - Tumor pharmacodynamic analysis reveals sustained CDK12 degradation (≥70%) in tumor tissue for 24 h after a single oral dose of 30 mg/kg [1] |
| Enzyme Assay |
CDK12 kinase activity assay: Recombinant CDK12/cyclin K complex is incubated with a fluorescently labeled peptide substrate and ATP in the presence of serial dilutions of BSJ-4-116. After incubation at 30°C for 60 min, kinase activity is measured by detecting phosphorylated substrate using a microplate reader. The assay determines that BSJ-4-116 does not directly inhibit CDK12 kinase activity (IC50 > 1 μM), confirming its action as a degrader rather than a direct inhibitor [1]
- Target engagement assay (AlphaLISA): Biotinylated CDK12 protein is immobilized on streptavidin-coated plates, and a europium-labeled antibody against CDK12 is added. Serial dilutions of BSJ-4-116 are incubated with the mixture, and target engagement is measured by detecting changes in AlphaLISA signal. The assay shows dose-dependent binding of BSJ-4-116 to CDK12 with a Kd of 3.7 ± 0.5 nM [1] |
| Cell Assay |
Cell Proliferation Assay[1]
Cell Types: Parental Kelly and CDK12C1039F Cell Tested Concentrations: 10-10000 nM Incubation Duration: 72 hrs (hours) Experimental Results: The anti-proliferative activity of BSJ-4-116 is independent of mutation status and the degrader compounds exhibit improved GR50 (Growth rate inhibition of Kelly CDK12C1039F cells compared to the parental cell line) value. Western Blot Analysis[1] Cell Types: Parental and Kelly cells expressing CDK12C1039F (KellyCDK12CF) Tested Concentrations: 50 nM Incubation Duration: 6-24 hrs (hours) Experimental Results: Result in the same level of reduction in CDK12 protein levels regardless of mutational status of the cell line. CDK12 degradation Western blot assay: Cancer cells are seeded in 6-well plates and cultured overnight. Cells are treated with serial dilutions of BSJ-4-116 (0.1–100 nM) for 24 h, then lysed in RIPA buffer. Lysates are separated by SDS-PAGE, transferred to PVDF membranes, and probed with antibodies against CDK12, CDK13, and GAPDH (loading control). Band intensity is quantified using densitometry to calculate DC50 values [1] - Cell proliferation assay: Cells are plated in 96-well plates (5 × 103 cells/well) and treated with BSJ-4-116 (0.01–1000 nM) for 72 h. Cell viability is measured by adding a tetrazolium salt reagent, incubating for 4 h, and reading absorbance at 490 nm. IC50 values are calculated using nonlinear regression analysis [1] - RT-qPCR for DDR gene expression: After treating cells with BSJ-4-116 (10 nM) for 24 h, total RNA is extracted and reverse-transcribed to cDNA. Quantitative PCR is performed using specific primers for BRCA1, BRCA2, ATM, and GAPDH (housekeeping gene). Relative gene expression is calculated using the ΔΔCt method [1] - Apoptosis assay: Cells are treated with BSJ-4-116 (100 nM) for 48 h, harvested, and stained with Annexin V-FITC and propidium iodide (PI) for 15 min in the dark. Apoptotic cells are analyzed by flow cytometry, with Annexin V+/PI- (early apoptosis) and Annexin V+/PI+ (late apoptosis) cells counted together [1] |
| Animal Protocol |
Xenograft tumor model establishment: Female nude mice (6–8 weeks old) are subcutaneously injected with 5 × 106 MDA-MB-231 or HCT116 cells suspended in Matrigel (1:1 v/v with PBS) into the right flank. Tumors are allowed to grow to 100–150 mm3 before initiating treatment [1]
- Drug formulation and administration: BSJ-4-116 is dissolved in a vehicle consisting of PEG400/ethanol/PBS (30:10:60 v/v/v). Mice are randomly divided into vehicle control and treatment groups (n = 6 per group). The treatment group receives oral gavage of BSJ-4-116 at 30 mg/kg once daily for 21 days, while the control group receives the vehicle alone [1] - Tumor and tissue collection: Tumor volume is measured every 3 days using calipers (volume = length × width2 / 2). At the end of the study, mice are euthanized, and tumors are excised, weighed, and snap-frozen in liquid nitrogen for Western blot and RT-qPCR analysis. Major organs (liver, kidney, spleen) are collected for histopathological examination [1] |
| ADME/Pharmacokinetics |
In mice, the bioavailability of BSJ-4-116 (30 mg/kg) orally was 42 ± 6%, with a peak plasma concentration (Cmax) of 1.8 ± 0.3 μM 1 hour after administration [1]
- The plasma half-life (t1/2) was 3.2 ± 0.5 hours, and the area under the plasma concentration-time curve (AUC0–24h) was 8.7 ± 1.2 μM·h [1] - Tissue distribution analysis showed that the drug accumulated highly in tumor tissues (tumor/plasma ratio of 3.8 ± 0.7), with moderate distribution in the liver and kidneys, and low concentrations in the brain and muscle [1] |
| Toxicity/Toxicokinetics |
In a 21-day xenotransplantation study, BSJ-4-116 (30 mg/kg, orally, once daily) did not cause significant weight loss (maximum weight loss of 4.2% compared to the control group), and no significant pathological changes were observed in the liver, kidneys, spleen, heart, or lungs [1]. The plasma protein binding rate in mouse plasma was 92 ± 3% as determined by balanced dialysis [1]. No significant changes were observed in serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), or creatinine levels, indicating no significant hepatotoxicity or nephrotoxicity was observed [1].
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| References | |
| Additional Infomation |
BSJ-4-116 is a proteolytic targeting chimera (PROTAC) that recruits the E3 ubiquitin ligase cereblon (CRBN) to mediate CDK12 ubiquitination and proteasome degradation [1]. - Resistance mechanism: Acquired resistance to BSJ-4-116 in cancer cells is associated with missense mutations in CDK12 (e.g., Cys1039Arg, Gly1042Asp), which disrupt the binding of PROTAC to CDK12, thereby blocking CDK12 degradation [1]. - In CDK12-dependent cancer cell lines and xenograft models, BSJ-4-116 exhibits synergistic antitumor activity with the PARP inhibitor (olaparib) because downregulation of the DDR gene by BSJ-4-116 enhances sensitivity to PARP. Inhibition [1]
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| Molecular Formula |
C40H49CLN8O8S
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|---|---|
| Molecular Weight |
837.3839
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| Exact Mass |
836.31
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| Elemental Analysis |
C, 57.37; H, 5.90; Cl, 4.23; N, 13.38; O, 15.28; S, 3.83
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| CAS # |
2519823-34-6
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| PubChem CID |
155235839
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| Appearance |
White to off-white solid powder
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| LogP |
5.4
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
13
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| Rotatable Bond Count |
18
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| Heavy Atom Count |
58
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| Complexity |
1570
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC(C)S(=O)(=O)C1=CC=CC=C1NC2=NC(=NC=C2Cl)N[C@@H]3CCCN(C3)CCCCCCCNC(=O)COC4=CC=CC5=C4C(=O)N(C5=O)C6CCC(=O)NC6=O
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| InChi Key |
YJOJMGTVKMABKQ-FIQOPJFZSA-N
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| InChi Code |
InChI=1S/C40H49ClN8O8S/c1-25(2)58(55,56)32-16-7-6-14-29(32)45-36-28(41)22-43-40(47-36)44-26-12-11-21-48(23-26)20-9-5-3-4-8-19-42-34(51)24-57-31-15-10-13-27-35(31)39(54)49(38(27)53)30-17-18-33(50)46-37(30)52/h6-7,10,13-16,22,25-26,30H,3-5,8-9,11-12,17-21,23-24H2,1-2H3,(H,42,51)(H,46,50,52)(H2,43,44,45,47)/t26-,30?/m1/s1
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| Chemical Name |
Acetamide, N-[7-[(3R)-3-[[5-chloro-4-[[2-[(1-methylethyl)sulfonyl]phenyl]amino]-2-pyrimidinyl]amino]-1-piperidinyl]heptyl]-2-[[2-(2,6-dioxo-3-piperidinyl)-2,3-dihydro-1,3-dioxo-1H-isoindol-4-yl]oxy]-
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| Synonyms |
BSJ4-116BSJ-4-116 BSJ 4-116BSJ4116 BSJ-4116BSJ 4116
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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 : ~250 mg/mL (~298.55 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (2.48 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 20.8 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: ≥ 2.08 mg/mL (2.48 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.1942 mL | 5.9710 mL | 11.9420 mL | |
| 5 mM | 0.2388 mL | 1.1942 mL | 2.3884 mL | |
| 10 mM | 0.1194 mL | 0.5971 mL | 1.1942 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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