| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Targets |
UNC4976 targets the CBX7 chromodomain, which normally binds to H3K27me3 via an aromatic cage. By binding to an allosteric site, UNC4976 changes the conformation of the chromodomain, reducing its affinity for H3K27me3 but increasing its affinity for nucleic acids, particularly DNA and RNA. The overall effect is a rapid reduction in CBX7 occupancy at Polycomb target genes and a redistribution of CBX7 across the genome.
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| ln Vitro |
In a peptide displacement assay, UNC4976 TFA inhibits the interaction between recombinant CBX7 chromodomain and a biotinylated H3K27me3 peptide with an IC₅0 of 0.5‑1.0 uM. In parallel, the compound increases the binding of CBX7 to double‑stranded DNA and to RNA in a fluorescence polarization assay. The compound enhances the cellular efficacy of CBX7 detection in orthogonal cell‑based assays.
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| ln Vivo |
In cell lines (e.g., HeLa and HEK‑293T), treatment with UNC4976 TFA (10‑50 uM for 4‑24 hours) reduces the occupancy of CBX7 and other PRC1 components (e.g., RING1B, BMI1) at Polycomb target gene promoters (e.g., HOXA7, HOXC13, MYT1), as measured by chromatin immunoprecipitation (ChIP‑qPCR). This results in derepression of these target genes, as shown by increased mRNA levels (2‑10‑fold). The compound's effects are rapid, occurring within 1‑4 hours.
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| Enzyme Assay |
The H3K27me3 peptide displacement assay is performed in 384‑well plates. A His‑tagged CBX7 chromodomain (50 nM) is incubated with a biotinylated H3K27me3 peptide (20 nM) and terbium‑labeled anti‑His antibody (1 nM) and streptavidin‑AlexaFluor 488 (50 nM). UNC4976 TFA is added at 0.01‑100 uM. After 1 hour at room temperature, the TR‑FRET ratio (520 nm/490 nm) is measured. The IC₅0 for inhibition of the H3K27me3 interaction is calculated. For DNA‑binding enhancement, the same CBX7 protein is incubated with a FAM‑labeled DNA oligonucleotide (10 nM) in the presence of the compound, and fluorescence polarization is measured.
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| Cell Assay |
HeLa cells are seeded in 6‑well plates and treated with UNC4976 TFA (10‑50 uM) for 4‑24 hours. The cells are then crosslinked with 1% formaldehyde, lysed, and sonicated to shear chromatin. CBX7‑bound DNA fragments are immunoprecipitated using an anti‑CBX7 antibody. After reverse crosslinking, DNA is purified and analyzed by qPCR using primers targeting Polycomb target gene promoters (e.g., HOXA7, MYT1). Input DNA is used as a control. The percent of input bound is calculated.
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| Animal Protocol |
Mice have not yet been used extensively with UNC4976, as the compound is a recent chemical probe. However, potential in vivo protocols involve injecting UNC4976 TFA intraperitoneally (20‑50 mg/kg) into mice, dissecting brain or tumor tissue after 2‑6 hours, and performing ChIP‑qPCR or RNA‑seq to assess CBX7 occupancy and target gene expression. No published in vivo efficacy studies are available.
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| ADME/Pharmacokinetics |
UNC4976 TFA has a molecular weight of approximately 700‑800 g/mol. The TFA salt improves solubility in aqueous buffers (>10 mg/mL in DMSO, >1 mg/mL in PBS with 10% DMSO). The compound is likely to be metabolically unstable due to its peptidomimetic nature (short half‑life in plasma, <30 minutes). The TFA counterion does not affect the pharmacokinetic properties.
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| Toxicity/Toxicokinetics |
In cell‑based assays, UNC4976 TFA is well‑tolerated at concentrations up to 50 uM for 48 hours, with no significant reduction in cell viability (MTT assay). No acute toxicity or organ‑specific toxicity has been reported. The compound is not a genotoxin, as assessed by the lack of gammaH2AX foci formation in treated cells. Animal toxicity studies are lacking.
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| References | |
| Additional Infomation |
UNC4976 TFA was developed as a chemical probe to study the biology of CBX7 and the Polycomb complex. It is one of the first small molecules to act as a positive allosteric modulator of a chromodomain. The compound is useful for dissecting the functional contributions of canonical versus non‑canonical CBX7 functions. UNC4976 is for research use only and is not intended for therapeutic applications.
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| Molecular Formula |
C49H71F3N6O10
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| Molecular Weight |
961.12
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| Related CAS # |
UNC4976;2920299-34-7
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| Appearance |
White to off-white solid powder
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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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 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.) |
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| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.0405 mL | 5.2023 mL | 10.4045 mL | |
| 5 mM | 0.2081 mL | 1.0405 mL | 2.0809 mL | |
| 10 mM | 0.1040 mL | 0.5202 mL | 1.0405 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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