BSJ-03-204 is a PROTAC linked by Cereblon ligand and CDK ligand. It is a potent and specific CDK4/6 dual degrader based on Palbociclib , targeting CDK4/D1 and CDK6/D1 The IC50 are 26.9 nM and 10.4 nM respectively. BSJ-03-204 does not induce IKZF1/3 degradation and has anti-neoplastic activity.

Physicochemical Properties

Molecular Formula	C43H48N10O8
Molecular Weight	832.903429031372
Exact Mass	832.365
CAS #	2349356-09-6
PubChem CID	139593561
Appearance	Light yellow to green yellow solid powder
LogP	2.2
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	14
Rotatable Bond Count	14
Heavy Atom Count	61
Complexity	1750
Defined Atom Stereocenter Count	0
SMILES	O=C1C(C(C)=O)=C(C)C2=CN=C(NC3=CC=C(C=N3)N3CCN(CCCCNC(COC4=CC=CC5C(N(C(C=54)=O)C4C(NC(CC4)=O)=O)=O)=O)CC3)N=C2N1C1CCCC1
InChi Key	KDMOCOWXLQOXEB-UHFFFAOYSA-N
InChi Code	InChI=1S/C43H48N10O8/c1-25-30-23-46-43(49-38(30)52(27-8-3-4-9-27)41(59)36(25)26(2)54)47-33-14-12-28(22-45-33)51-20-18-50(19-21-51)17-6-5-16-44-35(56)24-61-32-11-7-10-29-37(32)42(60)53(40(29)58)31-13-15-34(55)48-39(31)57/h7,10-12,14,22-23,27,31H,3-6,8-9,13,15-21,24H2,1-2H3,(H,44,56)(H,48,55,57)(H,45,46,47,49)
Chemical Name	N-[4-[4-[6-[(6-acetyl-8-cyclopentyl-5-methyl-7-oxopyrido[2,3-d]pyrimidin-2-yl)amino]pyridin-3-yl]piperazin-1-yl]butyl]-2-[2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl]oxyacetamide
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 Note: This product requires protection from light (avoid light exposure) during transportation and storage.
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

Targets	CDK4/D1 26.9 nM (IC50) CDK6/D1 10.4 nM (IC50)
ln Vitro	On MCL cell lines, BSJ-03-204 (0.0001-100 μM; for 3 or 4 days) has strong anti-proliferative effects[1]. A G1 arrest can be potently induced by BSJ-03-204 (1 μM; for 1 day)[1]. In WT cells, BSJ-03-204 (0.1–5 μM) solely degrades CDK4/6; it does not affect IKZF1/3[1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: MCL cell lines (Jeko-1, Mino, Granta-519, Rec-1, and Maver-1 cells) Tested Concentrations: 0.0001- 100 μM Incubation Duration: For 3 or 4 days Experimental Results: Had potent anti-proliferative effects on mantle cell lymphoma (MCL) cell lines. Cell Cycle Analysis[1] Cell Types: Granta-519 cells Tested Concentrations: 1 μM Incubation Duration: For 1 day Experimental Results: Potently induced a G1 arrest. Western Blot Analysis[1] Cell Types: Wildtype (WT) or Crbn-/- Jurkat cells Tested Concentrations: 0.1, 0.5, 1, 5 μM Incubation Duration: For 4 hrs (hours) Experimental Results: Only resulted in Degradation of CDK4/6 in WT cells, not IKZF1/3.
References	[1]. Development of Dual and Selective Degraders of Cyclin-Dependent Kinases 4 and 6. Angew Chem Int Ed Engl. 2019 May 6;58(19):6321-6326.

Solubility Data

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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.2006 mL	6.0031 mL	12.0062 mL
	5 mM	0.2401 mL	1.2006 mL	2.4012 mL
	10 mM	0.1201 mL	0.6003 mL	1.2006 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.