| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
SPEN (Split ends) protein, specifically binding to the RepA domain within the Xist long non-coding RNA. SPEN-IN-1 binds to RepA with high selectivity, disrupting the interaction between Xist and the polycomb repressive complex 2 (PRC2). The compound also inhibits RepA, PRC2, and SPEN, with Kd values of 400, 47, and 420 nM, respectively.
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| ln Vitro |
SPEN-IN-1 (0-100 μM) inhibits RepA-PRC2 and RepA-SPEN with IC50 values of 30 and 48 μM, respectively[1]. At Kd values of 400, 47, and 420 nM, respectively, SPEN-IN-1 inhibits RepA, PRC2, and SPEN[1]. In vitro and in cells, SPEN-IN-1 specifically ruins the connection between Xist and homologous proteins [1]. The accumulation of PRC2 and H3K27me3 is significantly inhibited by SPEN-IN-1 [1].
In vitro, SPEN-IN-1 selectively inhibits the SPEN-RNA interaction, disrupting the formation of the Xist-PRC2 complex. By blocking this interaction, it interferes with the initiation of X-chromosome inactivation. This compound is used as a molecular tool in cellular assays to dissect the mechanism of gene silencing on the inactive X chromosome and to identify SPEN-dependent gene regulation pathways. |
| ln Vivo |
In vivo, SPEN-IN-1 diminishes the RepA-SPEN interaction [1].
In vivo, SPEN-IN-1 serves as a valuable chemical probe to study the role of SPEN in animal models. While its primary use is in in vitro cell culture systems, it can be used in vivo to investigate the physiological consequences of acute SPEN inhibition on X-chromosome inactivation and other SPEN-mediated processes. It helps validate SPEN as a potential therapeutic target for diseases such as certain cancers. |
| Enzyme Assay |
Non-cell-based binding assays (SPR) are used to determine the interaction. Recombinant SPEN protein is immobilized on a chip. Increasing concentrations of SPEN-IN-1 (0-1000 nM) are injected over the chip. The binding affinity is measured by SPR. For selectivity, a fluorescence polarization assay with a labeled RNA probe representing the RepA motif can be used. The compound's ability to displace the probe is measured.
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| Cell Assay |
Mouse embryonic fibroblasts (MEFs) or human cell lines are treated with SPEN-IN-1 at concentrations of 0.1-10 uM for 24-72 hours. The effect on Xist-mediated silencing is determined by measuring the expression of X-linked genes via RT-qPCR. The efficacy of target engagement is confirmed by performing RNA immunoprecipitation (RIP) of Xist and SPEN, followed by RT-qPCR to quantify their interaction.
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| Animal Protocol |
X-chromosome inactivation models in mice can be used. Pregnant female mice are injected intraperitoneally with SPEN-IN-1 at doses of 10-30 mg/kg during critical stages of embryogenesis. Alternatively, adult mice are treated similarly to assess the impact on X-linked gene expression in specific tissues. Gene expression changes are evaluated by RNA-seq or RT-qPCR on isolated tissues to assess the compound's in vivo activity.
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| ADME/Pharmacokinetics |
Detailed pharmacokinetic data is not available for this research tool. As a small molecule (MW 416.47), it is expected to have good cell permeability and is orally bioavailable. SPEN-IN-1 shows excellent solubility in DMSO (66.67 mg/mL). For in vivo formulation, vehicles such as PEG300, Tween-80, and saline are typically used for intraperitoneal or oral administration.
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| Toxicity/Toxicokinetics |
SPEN-IN-1 is for research use only and has not been approved for human use. Acute toxicity studies have not been published, but as a chemical probe, it is generally used at concentrations (e.g., 0.1-10 uM in vitro) that are not overtly cytotoxic. At high concentrations, potential off-target effects on other RNA-binding proteins or transcription factors cannot be ruled out. Standard handling precautions should be taken.
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| References | |
| Additional Infomation |
SPEN-IN-1 is a research-grade chemical probe designed for epigenetic research. It has not entered clinical trials and is not a drug candidate. It is a valuable tool for exploring the fundamental biology of X-chromosome inactivation and the SPEN protein. This compound is not for therapeutic use and should be stored at -20degC in a dry, dark environment.
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| Molecular Formula |
C27H20N4O
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| Molecular Weight |
416.47
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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 (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 :~66.67 mg/mL (~160.08 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.00 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 (6.00 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 25.0 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 | 2.4011 mL | 12.0057 mL | 24.0113 mL | |
| 5 mM | 0.4802 mL | 2.4011 mL | 4.8023 mL | |
| 10 mM | 0.2401 mL | 1.2006 mL | 2.4011 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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