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Purity: ≥98%
BI-3812 (BI3812) is novel, highly potent and efficacious BCL6 inhibitor with potential antitumor activity. It inhibits the BTB domain of BCL6 with an IC50 of ≤3 nM. The transcription factor BCL6 is a known driver of oncogenesis in lymphoid malignancies, including diffuse large B cell lymphoma (DLBCL). Disruption of its interaction with transcriptional repressors interferes with the oncogenic effects of BCL6. This work establishes the BTB domain as a highly druggable structure, paving the way for the use of other members of this protein family as drug targets. The magnitude of effects elicited by this class of BCL6-degrading compounds exceeds that of our equipotent non-degrading inhibitors, suggesting opportunities for the development of BCL6-based lymphoma therapeutics.
| Targets |
BI-3812 is a potent inhibitor of the BTB (Broad-Complex, Tramtrack, and Bric a brac) domain of the oncogenic transcription factor BCL6 (B-cell lymphoma 6) [1].
BCL6 BTB domain (IC₅₀ ≤ 3 nM in ULight biochemical assay; cellular binding IC₅₀ = 40 nM in LUMIER assay) [1] |
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| ln Vitro |
BI-3812 is a highly potent and efficacious BCL6 inhibitor probe compound. It effectively inhibits the interaction between the BCL6 BTB domain and transcriptional co-repressors (e.g., NCOR1) in cellular assays (LUMIER), with an IC₅₀ of 40 nM. Unlike the degrader compound BI-3802, BI-3812 does not induce ubiquitination and proteasome-dependent degradation of BCL6 protein in various diffuse large B cell lymphoma (DLBCL) cell lines (SU-DHL-4, Farage, OCI-Ly1, BJAB). It is classified as a "non-degrader," causing minimal reduction (<30%) in BCL6 protein levels even at high concentrations. In transcriptional studies, treatment with BI-3812 induced de-repression of known BCL6 target genes (e.g., PRDM1, IRF4, PTPN6) in DLBCL cell lines, but the magnitude of induction was consistently weaker compared to the degrader BI-3802. In long-term proliferation assays across several DLBCL cell lines (e.g., SU-DHL-4, Farage, OCI-Ly7, OCI-Ly1), BI-3812 showed only marginal anti-proliferative effects, which were observable only at concentrations far above its biochemical and cellular IC₅₀ values for BCL6 inhibition. Cells not expressing BCL6, such as Toledo cells, were insensitive to BI-3812. The structurally related but low-affinity BCL6 binder BI-5273 had no effect on proliferation, confirming the on-target activity of BI-3812 [1].
Hydrogen deuterium exchange mass spectrometry (HDX MS) analysis indicated that binding of BI-3812 to the BCL6 BTB domain protected regions close to the binding site from deuterium exchange, but showed less protection in remote regions compared to the degrader BI-3802, suggesting differences in protein stabilization effects [1]. A chemoaffinity pulldown experiment with an immobilized analog of the degrader BI-3802 identified BCL6 as the major target in DLBCL cells; no other BTB/POZ domain-containing proteins were identified, suggesting selectivity within this protein family for the related compound series [1] |
| ln Vivo |
BCL6 reagent BI-3812 has an IC50 of less than 3 nM. For the inhibition of BCL6, BI-3812 has an IC50 value of 40 nM[1].
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| Cell Assay |
BCL6 degradation was quantified in SU-DHL-4 cells. Cells were suspended and treated with compounds at logarithmic dose series using a digital dispenser. After 90 minutes of incubation, cells were lysed. BCL6 protein levels in the lysates were analyzed using capillary electrophoresis (Wes system) with anti-BCL6 and anti-GAPDH antibodies for normalization, generating dose-response curves and DC₅₀ (degradation potency) values [1].
Long-term proliferation assays were performed in DLBCL cell lines. Cells were seeded and maintained at constant compound concentrations in multi-well plates. They were passaged periodically to maintain density, and cumulative cell numbers were calculated by multiplying split ratios over time to generate growth curves [1]. Cellular target engagement was measured using LUMIER (luminescence-based mammalian interactome) assays. HEK293T cells were co-transfected with expression constructs for Renilla luciferase-tagged BCL6 (containing the BTB domain) and Firefly luciferase-tagged co-repressor NCOR1. After compound treatment, cells were lysed, and the lysate was subjected to immunoprecipitation using anti-Renilla antibody. The co-precipitated Firefly luciferase activity (representing the BCL6-NCOR1 interaction) was measured and normalized to the Renilla luciferase activity in the input lysate [1]. For qPCR analysis of gene expression, cells were treated in 96-well plates, lysed, and mRNA levels were quantified using reverse transcription-quantitative PCR with gene-specific primers. Data were normalized to GAPDH mRNA levels [1]. To study domain requirements for degradation, various mutant and chimeric constructs of BCL6 (e.g., lacking the zinc finger domain or fused to heterologous DNA-binding domains) were transiently transfected into HEK293 cells. Protein levels were assessed by western blotting after compound treatment [1] . |
| References | |
| Additional Infomation |
BI-3812 is a highly effective and potent BCL6 inhibitor probe compound. Its structure is very similar to that of the degrader BI-3802, but its function is that of a "non-degrader," which allows researchers to distinguish the biological effects caused by BCL6 inhibition and BCL6 protein degradation. Studies have shown that although both inhibitor and degrader compounds can relieve the inhibition of a similar set of BCL6 target genes, the degrader has a significantly stronger antiproliferative effect in diffuse large B-cell lymphoma (DLBCL) cell lines. BI-3812 provides a tool for studying the specific consequences of disrupting the interaction of BCL6 co-inhibitors by inhibiting the BTB domain without causing degradation [1]. This study establishes the BCL6 BTB domain as a target with great drug potential. The discovery of potent, non-degrading inhibitors such as BI-3812 and degrading agents such as BI-3802 from the same chemical family provides new avenues for studying BCL6 biology and developing potential lymphoma treatments [1].
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| Molecular Formula |
C26H32CLN7O5
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|---|---|
| Molecular Weight |
558.029184341431
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| Exact Mass |
557.215
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| CAS # |
2166387-64-8
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| PubChem CID |
134691741
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| Appearance |
Off-white to pink solid powder
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| LogP |
2.2
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
39
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| Complexity |
928
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| Defined Atom Stereocenter Count |
0
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| SMILES |
ClC1C=NC(=NC=1NC1C=C(C2=C(C=C(C(N2C)=O)OCC(NC)=O)C=1)OC)N1CCC(C(N(C)C)=O)CC1
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| InChi Key |
XCGYXEVLQIIEJH-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C26H32ClN7O5/c1-28-21(35)14-39-20-11-16-10-17(12-19(38-5)22(16)33(4)25(20)37)30-23-18(27)13-29-26(31-23)34-8-6-15(7-9-34)24(36)32(2)3/h10-13,15H,6-9,14H2,1-5H3,(H,28,35)(H,29,30,31)
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| Chemical Name |
1-(5-Chloro-4-((8-methoxy-1-methyl-3-(2-(methylamino)-2-oxoethoxy)-2-oxo-1,2-dihydroquinolin-6-yl)amino)pyrimidin-2-yl)-N,N-dimethylpiperidine-4-carboxamide
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| Synonyms |
BI3812; BI 3812; BI-3812
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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 : ~20.83 mg/mL (~37.33 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.08 mg/mL (3.73 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
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 (3.73 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 | 1.7920 mL | 8.9601 mL | 17.9202 mL | |
| 5 mM | 0.3584 mL | 1.7920 mL | 3.5840 mL | |
| 10 mM | 0.1792 mL | 0.8960 mL | 1.7920 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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