| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
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| Targets |
BRD4
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|---|---|
| ln Vitro |
Cells co-transfected with FKBPF36V-mEGFP and mCherry-BRD4 showed rapid translocation of FKBPF36V-EGFP to the nucleus following addition of NICE-01 (200 nM, 40 min; Figure 1c). The combination of AP1867 and JQ1 was unable to induce this effect (Figure S1). We then hypothesized that endogenous BET-containing proteins could be used as “carriers” for nuclear import. In cells stably expressing FKBPF36V-mEGFP, NICE-01 (250 nM, 3 h) enriched mEGFP in the nucleus (Figure 1d and Figure S1). This effect could be complemented by JQ1 or AP1867 at high concentrations (10 μM, Figure 1e).[1]
n cells lacking exogenous mCherry-BRD4, 10 μM NICE-01 was unable to induce nuclear import of FKBPF36V-mEGFP. [1] NICE-01 was able to import FKBPF36V-mEGFP-PIK3CAE545K in cells co-transfected with mCherry-BRD4.[1] Nuclear import showed first-order kinetics with respect to the FKBPF36V-mEGFP fraction in the cytoplasm after a short initial period (during which nuclear FKBPF36V-mEGFP (39.4 kDa) forms a ternary complex with mCherry-BRD4 and NICE-01); kimport was estimated to be 0.0266 min–1, and t1/2 = 26 min.[1] Cells co-transfected with FKBPF36V-mEGFP-NPM1c and mCherry-BRD4 showed rapid relocalization of FKBPF36V-mEGFP-NPM1c into the nucleus within minutes upon treatment with NICE-01 (Figure 2a,b; P = 4.4e-185 for the Mann–Whitney U test between distribution of cellular mEGFP-mCherry correlation before compound addition and 25 min after NICE-01 addition).[1] |
| Cell Assay |
Rapid import of GFP into the nucleus by AP1867-PEG2-JQ1 (NICE-01). (A) Schematic of the Nuclear Import and Control of Expression (NICE) concept. A bifunctional molecule allows import of a cytosolic protein of interest into the nucleus. (B) Structure of AP1867-PEG2-JQ1. (C) U2OS cells co-transfected with mCherry-BRD4 and FKBPF36V-mEGFP, treated with NICE-01 (200 nM) or DMSO, and imaged for 40 min. (D) 293T cells stably expressing FKBPF36V-mEGFP, treated with NICE-01 (250 nM) or DMSO, and imaged for 3 h. (E) 293T cells stably expressing FKBPF36V-mEGFP and co-treated with NICE-01 (250 nM) and either JQ1 or AP1867 at the indicated concentration and imaged for 15 min. (F) 293T cells stably expressing FKBPF36V-mEGFP and transfected with mCherry-BRD4, and either untreated or treated with NICE-01 (250 nM, 10 μM), and imaged for 15 min. White arrows indicate representative cells exhibiting the hook effect. Red arrow indicates representative cell with mCherry-BRD4 expression that does not exhibit hook effect. (G) 293T cells stably expressing FKBPF36V-mEGFP and transfected with mCherry-BRD4 and treated with NICE-01 (250 nM) and imaged for 50 min. 100 mCherry+ and mCherry– cells were randomly selected and assessed for complete import of mEGFP (cellular nuclear/cytosolic mEGFP intensity ≥2.5). P values were calculated by Fisher’s exact test.[1]
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| References |
[1]. Bifunctional small molecules that induce nuclear localization and targeted transcriptional regulation. J Am Chem Soc . 2023 Dec 6;145(48):26028-26037. https://pubmed.ncbi.nlm.nih.gov/37992275/
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| Additional Infomation |
The aberrant localization of proteins in cells is a key factor in the development of various diseases, including cancer and neurodegenerative disease. To better understand and potentially manipulate protein localization for therapeutic purposes, we engineered bifunctional compounds that bind to proteins in separate cellular compartments. We show these compounds induce nuclear import of cytosolic cargoes, using nuclear-localized BRD4 as a "carrier" for co-import and nuclear trapping of cytosolic proteins. We use this system to calculate kinetic constants for passive diffusion across the nuclear pore and demonstrate single-cell heterogeneity in response to these bifunctional molecules with cells requiring high carrier to cargo expression for complete import. We also observe incorporation of cargo into BRD4-containing condensates. Proteins shown to be substrates for nuclear transport include oncogenic mutant nucleophosmin (NPM1c) and mutant PI3K catalytic subunit alpha (PIK3CAE545K), suggesting potential applications to cancer treatment. In addition, we demonstrate that chemically induced localization of BRD4 to cytosolic-localized DNA-binding proteins, namely, IRF1 with a nuclear export signal, induces target gene expression. These results suggest that induced localization of proteins with bifunctional molecules enables the rewiring of cell circuitry, with significant implications for disease therapy.
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| Molecular Formula |
C63H76CLN7O13S
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|---|---|
| Molecular Weight |
1206.83
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| Exact Mass |
1205.49103
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| CAS # |
2982819-94-1
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| PubChem CID |
169183208
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
8.8
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| SMILES |
CC[C@@H](C1=CC(=C(C(=C1)OC)OC)OC)C(=O)N2CCCC[C@H]2C(=O)O[C@H](CCC3=CC(=C(C=C3)OC)OC)C4=CC(=CC=C4)OCC(=O)NCCOCCOCCNC(=O)C[C@H]5C6=NN=C(N6C7=C(C(=C(S7)C)C)C(=N5)C8=CC=C(C=C8)Cl)C
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| InChi Key |
USCBOLCYUGPYAF-YUZLDYPQSA-N
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| InChi Code |
InChI=1S/C63H76ClN7O13S/c1-10-47(44-34-53(78-7)59(80-9)54(35-44)79-8)61(74)70-27-12-11-16-49(70)63(75)84-50(23-17-41-18-24-51(76-5)52(32-41)77-6)43-14-13-15-46(33-43)83-37-56(73)66-26-29-82-31-30-81-28-25-65-55(72)36-48-60-69-68-40(4)71(60)62-57(38(2)39(3)85-62)58(67-48)42-19-21-45(64)22-20-42/h13-15,18-22,24,32-35,47-50H,10-12,16-17,23,25-31,36-37H2,1-9H3,(H,65,72)(H,66,73)/t47-,48-,49-,50+/m0/s1
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| Chemical Name |
[(1R)-1-[3-[2-[2-[2-[2-[[2-[(9S)-7-(4-chlorophenyl)-4,5,13-trimethyl-3-thia-1,8,11,12-tetrazatricyclo[8.3.0.02,6]trideca-2(6),4,7,10,12-pentaen-9-yl]acetyl]amino]ethoxy]ethoxy]ethylamino]-2-oxoethoxy]phenyl]-3-(3,4-dimethoxyphenyl)propyl] (2S)-1-[(2S)-2-(3,4,5-trimethoxyphenyl)butanoyl]piperidine-2-carboxylate
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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 | 0.8286 mL | 4.1431 mL | 8.2862 mL | |
| 5 mM | 0.1657 mL | 0.8286 mL | 1.6572 mL | |
| 10 mM | 0.0829 mL | 0.4143 mL | 0.8286 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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