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Purity: ≥98%
BI-3802 (BI3802) is a novel and highly potent BCL6 degrader which inhibits the BTB domain of BCL6 with an IC50 of ≤3 nM; BI-3802 has demonstrated potent antitumor activity by inducing efficacious BCL6 protein degradation in several diffuse large B-cell lymphoma (DLBCL) cell lines.
| Targets |
BI-3802 is a potent and selective inhibitor of the BTB (Broad-Complex, Tramtrack, and Bric a brac) domain of the transcription factor BCL6 (B-cell lymphoma 6). It inhibits the interaction between the BCL6 BTB domain and transcriptional co-repressors (e.g., NCOR1, SMRT, BCOR) with an IC₅₀ ≤ 3 nM in a biochemical ULight assay. [1]
Additionally, BI-3802 induces rapid, proteasome-dependent degradation of BCL6 protein in cells, a property not solely dependent on its inhibitory potency. [1] |
|---|---|
| ln Vitro |
The cellular BCL6 IC50 of BI-3802 is 43 nM[1]. The reaction between BCL6 and SIAH1 is increased by BI-3802 (EC50=64 nM)[2].
BI-3802 induces rapid and potent degradation of BCL6 protein in multiple diffuse large B cell lymphoma (DLBCL) cell lines (e.g., SU-DHL-4, Farage, OCI-Ly1, BJAB), reducing BCL6 levels by ≥90% within 2 hours. The degradation half-life is approximately 5 minutes. [1] Degradation is mediated by the ubiquitin-proteasome system, as it is blocked by the proteasome inhibitor MG-132 and leads to the accumulation of poly-ubiquitylated BCL6. [1] BI-3802 causes strong de-repression of BCL6 target genes (e.g., PRDM1, IRF4, CD69, ATM, DUSP5) as measured by RNA-seq and qPCR, with effects significantly greater than those of a non-degrading but equipotent inhibitor (BI-3812). [1] BI-3802 exhibits anti-proliferative effects in several BCL6-positive DLBCL cell lines (e.g., SU-DHL-4, Farage, OCI-Ly1), leading to a slow-onset growth arrest over 4-7 days, but does not significantly induce apoptosis. BCL6-negative cell lines (e.g., Toledo, MV-4-11) are insensitive. [1] Chemoaffinity pulldown with an immobilized BI-3802 analog in Farage cell lysate identified BCL6 as the major target, with no other BTB/POZ domain-containing proteins detected, indicating high selectivity. [1] |
| Enzyme Assay |
A fluorescence polarization (FP) assay was used to monitor the inhibition of the interaction between a fluorescently labeled BCOR peptide and the recombinant BCL6 BTB domain protein. Compounds were screened in 384-well plates. The assay buffer contained HEPES, NaCl, Triton X-100, and reduced glutathione. After adding protein and peptide to compound-containing wells, plates were incubated and read on a fluorescence plate reader. [1]
A ULight time-resolved fluorescence resonance energy transfer (TR-FRET) assay was also used for more sensitive inhibition measurements. In this assay, biotinylated BCL6 BTB domain protein, a ULight-labeled BCOR peptide, and streptavidin-europium were used. Compounds were pre-incubated with the protein, followed by addition of the peptide and streptavidin-europium mix. After incubation, the TR-FRET signal was measured. A modified "ULight LOW" assay with lower protein concentration was used to accurately measure IC₅₀ values below 50 nM. [1] Surface plasmon resonance (SPR) was used to confirm direct, dose-dependent, and saturable binding of compounds to the BCL6 BTB domain and to determine dissociation constants (K_D). [1] |
| Cell Assay |
To quantify BCL6 degradation, SU-DHL-4 cells were suspended and treated with compound for 90 minutes. Cells were then lysed, and BCL6 protein levels were analyzed by capillary electrophoresis (Wes system) using specific antibodies, with GAPDH for normalization. Dose-response curves were generated to calculate DC₅₀ (concentration for half-maximal degradation) values. [1]
For long-term proliferation assays, DLBCL cell lines were seeded in 24-well plates at a defined density and maintained at constant compound concentrations. Cells were passaged every 3-4 days, and cumulative cell numbers were calculated by multiplying split ratios to generate growth curves over time. [1] Gene expression analysis was performed by RNA sequencing. Cells were treated with compound or DMSO for 20 hours or 7 days. For the 7-day treatment, cells were split and fresh compound was added at day 3. RNA was extracted, libraries were prepared from polyA-enriched RNA, and sequenced. Differential gene expression analysis was performed using bioinformatics tools. [1] Quantitative PCR (qPCR) was used to validate gene expression changes. Cells were treated, lysed, and cDNA was synthesized and amplified using gene-specific primers and a reference gene (GAPDH). Relative mRNA abundance was calculated. [1] For detection of poly-ubiquitylated BCL6, cells were treated with compound and proteasome inhibitor, followed by lysis and immunoprecipitation of BCL6. The immunoprecipitates were analyzed by western blotting using TUBEs (tandem ubiquitin-binding entities) to detect ubiquitin chains. [1] |
| References | |
| Additional Infomation |
BI-3802 is defined as a “degrader”, meaning that it not only inhibits the BTB domain of BCL6 but also induces its rapid ubiquitin-proteasome-dependent degradation. This contrasts with “non-degrader” inhibitors such as BI-3812, which only block the binding of co-repressors. [1] The degradation induced by BI-3802 requires DNA-binding ability of BCL6 (through its zinc finger domain or heteroDNA-binding domain) but does not require the known PEST domain, MAPK phosphorylation site, ATM/ATR pathway, or E3 ligase FBXO11. This suggests the existence of a novel degradation mechanism triggered by both compound binding and DNA binding. [1] Compared to BI-3812, BI-3802 exhibits stronger antiproliferative and derepressive effects, suggesting that complete removal of the BCL6 protein is more effective than simply inhibiting the function of its BTB domain, possibly due to the elimination of repressive functions mediated by other regions of BCL6. [1]
BI-3802 can be used as a highly efficient probe compound to study the effects of BCL6 degradation on cellular processes such as BCL6-dependent lymphoma and germinal center response. [1] |
| Molecular Formula |
C24H29CLN6O3
|
|---|---|
| Molecular Weight |
484.9785
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| Exact Mass |
484.198
|
| CAS # |
2166387-65-9
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| PubChem CID |
131632978
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| Appearance |
White to off-white solid powder
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| LogP |
4
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| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
6
|
| Heavy Atom Count |
34
|
| Complexity |
772
|
| Defined Atom Stereocenter Count |
2
|
| SMILES |
ClC1=CN=C(N=C1NC1C=CC2=C(C=C(C(N2C)=O)OCC(NC)=O)C=1)N1C[C@H](C)C[C@H](C)C1
|
| InChi Key |
GXTJETQFYHZHNB-GASCZTMLSA-N
|
| InChi Code |
InChI=1S/C24H29ClN6O3/c1-14-7-15(2)12-31(11-14)24-27-10-18(25)22(29-24)28-17-5-6-19-16(8-17)9-20(23(33)30(19)4)34-13-21(32)26-3/h5-6,8-10,14-15H,7,11-13H2,1-4H3,(H,26,32)(H,27,28,29)/t14-,15+
|
| Chemical Name |
2-((6-((5-Chloro-2-((3S,5R)-3,5-dimethylpiperidin-1-yl)pyrimidin-4-yl)amino)-1-methyl-2-oxo-1,2-dihydroquinolin-3-yl)oxy)-N-methylacetamide
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| Synonyms |
BI-3802; U52; BI 3802; U-52; BI3802; U 52
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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 : ~6.67 mg/mL (~13.75 mM)
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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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0619 mL | 10.3097 mL | 20.6194 mL | |
| 5 mM | 0.4124 mL | 2.0619 mL | 4.1239 mL | |
| 10 mM | 0.2062 mL | 1.0310 mL | 2.0619 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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