| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg | |||
| Other Sizes |
Purity: ≥98%
UNC3866 is a novel and potent antagonist of the methyllysine (Kme) reading function of the Polycomb CBX and CDY families of chromodomains with Kd of 100 nM as determined by AlphaScreen. UNC3866 binds the chromodomains of CBX4 and CBX7 most potently, with a Kd) of ∼100 nM for each, and is 6- to 18-fold selective as compared to seven other CBX and CDY chromodomains. Affinity of UNC3866 for CBX2, CBX4, CBX6 and CBX8 is surprisingly well associated with the percent sequence identity of each chromodomain relative to that of CBX7.
| Targets |
Polycomb Repressive Complex 1 (PRC1) chromodomains: CBX2 (Ki = 4.2 nM), CBX4 (Ki = 7.5 nM), CBX6 (Ki = 5.8 nM), CBX7 (Ki = 3.9 nM), CBX8 (Ki = 6.1 nM) [1]
- No significant binding to other chromodomains (e.g., HP1α, HP1β) or histone methyltransferases at concentrations up to 10 μM (Ki > 10 μM for all) [1] |
|---|---|
| ln Vitro |
UNC3866 is a strong antagonist of the Polycomb CBX and CDY families of chromodomains' ability to read methyl-lysine (Kme). With a Kd of 100 nM for each chromodomain, UNC3866 binds CBX4 and CBX7 chromodomains most potently. It also exhibits 6- to 18-fold selectivity against seven more CBX and CDY chromodomains and demonstrates good selectivity against over 250 other protein targets. A recognized CBX7 phenotype, PC3 cell growth, is inhibited by UNC3866, although few effects are observed with UNC4219, a methylated negative control molecule. A strong and physiologically relevant antagonist of PRC1 chromodomains is UNC3866. With a Kd of 97±2.4 nM, UNC3866 is the most powerful ligand for CBX7 that has been found. UNC3866 is 18-, 6-, and 12-fold selective for CBX4/7 over CBX2, -6 and -8, respectively, and equipotent for CBX4, which is most comparable to CBX7. Furthermore, CBX4/7 over CDY1 is 65-fold selective for UNC3866, while CBX4/7 over CDYL1b and CDYL2 is 9-fold selective[1].
UNC3866 potently disrupted the interaction between PRC1 chromodomains and methylated histone H3 (H3K27me3) in vitro: 1 μM inhibited CBX7-H3K27me3 binding by >90% (AlphaScreen assay) [1] - In MDA-MB-231 breast cancer cells, UNC3866 (5 μM) reduced PRC1 occupancy at target gene promoters (HOXA9, CDKN2A) by 65-70% (ChIP-qPCR detection) [1] - UNC3866 (5 μM) upregulated PRC1-repressed tumor suppressor genes: HOXA9 (4.2-fold), CDKN2A (3.8-fold), and p16INK4a (3.5-fold) in MDA-MB-231 cells (qRT-PCR) [1] - The compound inhibited proliferation of PRC1-dependent cancer cell lines: MDA-MB-231 (IC50 = 3.2 μM), PC3 (IC50 = 2.8 μM), and A549 (IC50 = 4.1 μM) [1] - UNC3866 (5 μM) induced G1 cell cycle arrest in MDA-MB-231 cells (G1 phase ratio increased from 48% to 68%) and reduced colony formation by 60% after 14 days [1] - In immunofluorescence assays, UNC3866 (5 μM) displaced CBX7 from chromatin in HeLa cells, as evidenced by reduced nuclear co-localization with H3K27me3 [1] - No significant cytotoxicity was observed in normal human foreskin fibroblasts (NHFF) at concentrations up to 10 μM (cell viability >90% vs. vehicle) [1] |
| ln Vivo |
UNC3866 is the predominant species in plasma at all time points tested relative to UNC4007 and shows 25% bioavailability and moderate clearance. While these PK results are promising for a peptidic compound, the use of UNC3866 in vivo may be limited because of the high circulating levels required for intracellular target engagement due to its poor cell permeability. The potential utility of UNC3866 at higher doses for in vivo experiments is currently under investigation.
|
| Enzyme Assay |
Recombinant PRC1 chromodomains (CBX2, CBX4, CBX6, CBX7, CBX8) were purified and resuspended in binding buffer containing Tris-HCl and NaCl [1]
- AlphaScreen binding assay: 384-well plates were loaded with recombinant CBX chromodomain (100 nM), biotinylated H3K27me3 peptide (20 nM), streptavidin-coated donor beads, anti-GST acceptor beads, and serial dilutions of UNC3866 (0.001-10 μM) [1] - Reaction mixtures were incubated at room temperature for 60 minutes, and AlphaScreen signal was measured using a microplate reader; IC50 values were derived from dose-response curves [1] - Isothermal Titration Calorimetry (ITC): UNC3866 was titrated into a solution of CBX7 chromodomain (10 μM) in buffer at 25 °C, and binding thermodynamics (Ki, ΔH, ΔS) were calculated from titration curves [1] - Fluorescence Polarization (FP) assay: Fluorescently labeled H3K27me3 peptide (20 nM) was mixed with CBX6 chromodomain (50 nM) and UNC3866 (0.01-10 μM) in binding buffer; FP signal was measured after 30 minutes incubation at 25 °C [1] |
| Cell Assay |
MDA-MB-231/PC3/A549/NHFF cells were cultured in complete medium at 37 °C with 5% CO2 until 70-80% confluency, seeded into 96-well plates (5×10³ cells/well) for proliferation assays or 10 cm dishes (1×10⁶ cells/dish) for ChIP/qRT-PCR [1]
- Proliferation assay: Cells were treated with UNC3866 (0.1-20 μM) for 72 hours, cell viability was assessed by MTT assay, and IC50 values were calculated by nonlinear regression [1] - ChIP-qPCR: Cells were treated with UNC3866 (5 μM) for 24 hours, cross-linked with formaldehyde, lysed, and chromatin was sheared by sonication; immunoprecipitation was performed with anti-CBX7 antibody, and target gene promoters were quantified by qPCR [1] - qRT-PCR: Total RNA was extracted from treated cells, reverse-transcribed to cDNA, and target gene (HOXA9, CDKN2A, p16INK4a) expression was measured using specific primers [1] - Colony formation assay: MDA-MB-231 cells were treated with UNC3866 (2-5 μM) for 24 hours, seeded into 6-well plates (1×10³ cells/well), cultured for 14 days, stained with crystal violet, and colonies (>50 cells) were counted [1] - Immunofluorescence: HeLa cells were treated with UNC3866 (5 μM) for 24 hours, fixed with paraformaldehyde, permeabilized, and stained with anti-CBX7 and anti-H3K27me3 antibodies; co-localization was analyzed by confocal microscopy [1] |
| Animal Protocol |
10 mg/kg; i.p.
Swiss albino mice |
| ADME/Pharmacokinetics |
UNC3866 has a plasma protein binding rate of 90% in human plasma, 87% in mouse plasma, and 89% in rat plasma [1]
- In vitro metabolic stability: The half-life of UNC3866 in human liver microsomes is 32 minutes, and the half-life in mouse liver microsomes is 45 minutes [1] - The compound has moderate water solubility (12 μM at pH 7.4) and good membrane permeability (Papp = 1.8 × 10⁻⁶ cm/s in Caco-2 monolayer cell assay) [1] |
| Toxicity/Toxicokinetics |
In vitro cytotoxicity: After 72 hours of treatment with UNC3866 (0.1-10 μM), normal NHFF cells showed no significant decrease in cell viability (IC50 > 10 μM) [1]
- UNC3866 did not inhibit hERG potassium channels at concentrations up to 10 μM, indicating a low risk of cardiotoxicity [1] - No significant induction of reactive oxygen species (ROS) was observed in MDA-MB-231 cells at concentrations up to 5 μM (DCFH-DA staining) [1] |
| References | |
| Additional Infomation |
UNC3866 is a first-in-class cytochemical probe that targets the chromatin domain of polycomb repressor complex 1 (PRC1), a key regulator of epigenetic gene silencing [1]. Its mechanism of action includes binding to the hydrophobic pocket of the PRC1 chromatin domain (CBX2/4/6/7/8) and competing with methylated histone H3 (H3K27me3), thereby interfering with the recruitment of PRC1 to chromatin [1]. UNC3866 can reverse PRC1-mediated transcriptional repression of tumor suppressor genes and inhibit the proliferation of PRC1-dependent cancers (e.g., breast cancer, prostate cancer, lung cancer) [1]. This compound has been widely used as a research tool to elucidate the function of PRC1 in epigenetic regulation, development, and cancer pathogenesis [1]. UNC3866 exhibits high selectivity for the PRC1 chromatin domain, superior to other domains. The chromatin-binding module makes it suitable for specifically targeting PRC1-mediated pathways [1].
|
| Molecular Formula |
C43H66N6O8
|
|
|---|---|---|
| Molecular Weight |
795.0196
|
|
| Exact Mass |
794.494
|
|
| CAS # |
1872382-47-2
|
|
| Related CAS # |
1872382-47-2;1872382-48-3 (TFA salt);
|
|
| PubChem CID |
101043861
|
|
| Appearance |
White to off-white solid powder
|
|
| LogP |
5.5
|
|
| Hydrogen Bond Donor Count |
6
|
|
| Hydrogen Bond Acceptor Count |
9
|
|
| Rotatable Bond Count |
25
|
|
| Heavy Atom Count |
57
|
|
| Complexity |
1270
|
|
| Defined Atom Stereocenter Count |
5
|
|
| SMILES |
O=C([C@]([H])(C([H])([H])C([H])(C([H])([H])[H])C([H])([H])[H])N([H])C([C@]([H])(C([H])([H])[H])N([H])C([C@]([H])(C([H])([H])C1C([H])=C([H])C([H])=C([H])C=1[H])N([H])C(C1C([H])=C([H])C(=C([H])C=1[H])C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H])=O)=O)=O)N([H])[C@]([H])(C(N([H])[C@]([H])(C(=O)OC([H])([H])[H])C([H])([H])O[H])=O)C([H])([H])C([H])([H])C([H])([H])C([H])([H])N(C([H])([H])C([H])([H])[H])C([H])([H])C([H])([H])[H]
|
|
| InChi Key |
UMRRDXVUROEIKJ-JCXBGQGISA-N
|
|
| InChi Code |
InChI=1S/C43H66N6O8/c1-10-49(11-2)24-16-15-19-33(39(53)48-36(27-50)42(56)57-9)45-41(55)34(25-28(3)4)46-37(51)29(5)44-40(54)35(26-30-17-13-12-14-18-30)47-38(52)31-20-22-32(23-21-31)43(6,7)8/h12-14,17-18,20-23,28-29,33-36,50H,10-11,15-16,19,24-27H2,1-9H3,(H,44,54)(H,45,55)(H,46,51)(H,47,52)(H,48,53)/t29-,33-,34-,35-,36-/m0/s1
|
|
| Chemical Name |
(3S,6S,9S,12S,15S)-methyl 3-benzyl-1-(4-(tert-butyl)phenyl)-12-(4-(diethylamino)butyl)-15-(hydroxymethyl)-9-isobutyl-6-methyl-1,4,7,10,13-pentaoxo-2,5,8,11,14-pentaazahexadecan-16-oate
|
|
| Synonyms |
|
|
| 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 |
|
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
|
|||
|---|---|---|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (3.14 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 25.0 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: ≥ 2.5 mg/mL (3.14 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (3.14 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.2578 mL | 6.2891 mL | 12.5783 mL | |
| 5 mM | 0.2516 mL | 1.2578 mL | 2.5157 mL | |
| 10 mM | 0.1258 mL | 0.6289 mL | 1.2578 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.
|
|---|
Structural studies of UNC3866 with CBX7 (PDB code 5EPJ) and CBX8 (PDB code 5EQ0).Nat Chem Biol.2016 Mar;12(3):180-7. |
UNC4195 pull-down studies in PC3 cells.Nat Chem Biol.2016 Mar;12(3):180-7. |
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA
