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| 5mg |
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| Targets |
DC-Y13-27 targets YTHDF2 (YTH domain family protein 2), a cytoplasmic and nuclear protein that specifically recognizes and binds to N6-methyladenosine (m6A)-modified RNA. YTHDF2 functions as an m6A reader that promotes degradation of methylated mRNA transcripts, thereby regulating gene expression post-transcriptionally. By binding to YTHDF2 (KD 37.9 microM) and inhibiting its RNA-binding activity, DC-Y13-27 stabilizes m6A-modified mRNA transcripts of its target genes, including FOXO3 (forkhead box O3) and TIMP1 (tissue inhibitor of metalloproteinases 1), and suppresses the expression of matrix metalloproteinases (MMP1, MMP3, MMP7, MMP9). DC-Y13-27 also induces pyroptosis, an inflammatory form of programmed cell death.
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| ln Vitro |
In vitro, DC-Y13-27 exhibits biological activity through inhibition of YTHDF2, with a KD of 37.9 uM. It upregulates the protein levels of FOXO3 and TIMP1, while downregulating MMP1, MMP3, MMP7, and MMP9, which are involved in extracellular matrix degradation. DC-Y13-27 also induces pyroptosis and increases IL-1beta secretion. In cancer research, DC-Y13-27 enhances the response to radiotherapy in colon cancer and melanoma cell lines, and has demonstrated antitumor activity against breast cancer cells. The compound reduces intervertebral disc degeneration by inhibiting YTHDF2-mediated catabolic gene expression, promoting matrix synthesis and reducing inflammatory responses in nucleus pulposus cells.
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| ln Vivo |
In vivo, DC-Y13-27 has shown efficacy in a variety of preclinical models. In the MC38 murine colon cancer model, DC-Y13-27 significantly enhanced tumor growth inhibition when used in combination with radiotherapy and/or anti-PD-L1 immune checkpoint therapy, via an increase in the number of CD8+ T cells within the tumor microenvironment. In murine models of breast cancer and melanoma, DC-Y13-27 in combination with radiotherapy led to a significant reduction in tumor growth and improved survival. Additionally, in a rat model of intervertebral disc degeneration (IVDD), administration of DC-Y13-27 reduced disc degeneration as assessed by histological scoring and radiographic analysis, and preserved disc height, indicating its potential as a disease-modifying therapy for IVDD.
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| Enzyme Assay |
The in vitro binding affinity between DC-Y13-27 and YTHDF2 is determined by surface plasmon resonance (SPR). Recombinant human YTHDF2 protein (the YTH domain or full-length) is immobilized onto a CM5 sensor chip via amine coupling. DC-Y13-27 is dissolved in DMSO and serially diluted in running buffer (PBS with 0.05% Tween-20, 1% DMSO) to final concentrations of 1-400 microM. The compound is flowed over the sensor chip at a flow rate of 30 microL/min for 120 seconds (association), followed by a dissociation phase of 300 seconds. The sensor chip is regenerated with 10 mM glycine-HCl (pH 2.0) between cycles. The resulting sensorgrams are fitted to a 1:1 Langmuir binding model using BIAevaluation software to calculate the association rate (ka), dissociation rate (kd), and equilibrium dissociation constant (KD). DC-Y13-27 binds YTHDF2 with a KD of 37.9 microM. Isothermal titration calorimetry (ITC) can also be used to confirm the binding thermodynamics and affinity. No specific enzyme-receptor binding protocols beyond SPR have been described for this compound.
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| Cell Assay |
Cellular assays for YTHDF2 inhibition are performed in human cancer cell lines, including HCT116 (colon cancer), A375 (melanoma), MDA-MB-231 (breast cancer), and rat primary nucleus pulposus cells (for IVDD). Cells are cultured in DMEM or RPMI-1640 medium with 10% FBS at 37degC and 5% CO2. DC-Y13-27 is dissolved in DMSO to a 50 mM stock, then diluted in culture medium to final concentrations of 1, 10, 25, 50, 100 uM (final DMSO ≤0.1%). For m6A-dependent gene regulation studies, cells are treated with DC-Y13-27 for 24-72 h. Total RNA is extracted with TRIzol and reverse transcribed. Quantitative RT-PCR is performed for FOXO3, TIMP1, MMP1, MMP3, MMP7, MMP9, and IL-1beta. For protein analysis, whole cell lysates are prepared, separated by SDS-PAGE, and Western blotted with antibodies against FOXO3, TIMP1, MMP1, MMP3, MMP7, MMP9, IL-1beta, cleaved GSDMD (gasdermin D; a marker of pyroptosis), and beta-actin as loading control. To assess pyroptosis, cells are treated with DC-Y13-27 for 24 h and then stained with propidium iodide (PI) and Annexin V. The percentage of PI-positive (necrotic/pyroptotic) cells is determined by flow cytometry. LDH release assay is also performed as a measure of plasma membrane damage, a hallmark of pyroptosis. Cell viability is assessed using MTT or CellTiter-Glo assays. To evaluate the effect of DC-Y13-27 on radiosensitivity, cells are pre-treated with DC-Y13-27 for 4 h, then exposed to increasing doses of ionizing radiation (2-10 Gy, using a cesium-137 or X-ray irradiator). After 7-10 days, colony formation assays are performed to calculate the surviving fraction and the radiosensitizing enhancement ratio (SER).
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| Animal Protocol |
In vivo efficacy is evaluated in the MC38 syngeneic colon carcinoma model in C57BL/6 mice. For tumor growth delay studies, female C57BL/6 mice (6-8 weeks) are injected subcutaneously with 1 × 10⁶ MC38 cells. When tumors reach ~100 mm3 (day 7-10), mice are randomized (n = 10/group). DC-Y13-27 is formulated in PBS with 5% DMSO and 10% Cremophor EL, or in 10% DMSO/40% PEG300/5% Tween-80/45% saline, and administered by intraperitoneal (i.p.) injection at doses of 25-100 mg/kg, once daily for 14-21 days. For radiotherapy, tumors are treated with 8-12 Gy local irradiation using a small animal irradiator (e.g., X-RAD 225Cx) either as a single fraction or 2 Gy × 5 fractions. For anti-PD-L1 combination studies, mice receive anti-mouse PD-L1 antibody (10 mg/kg, i.p., twice weekly). Tumor volume (length × width2/2) is measured every 2-3 days. The primary endpoint is tumor growth inhibition (TGI, %). At study termination, tumors are excised, weighed, and processed for flow cytometry of CD8+ T cells, CD4+ T cells, Tregs (FoxP3+), and for TUNEL staining to assess apoptosis. For the intervertebral disc degeneration (IVDD) model, male Sprague-Dawley rats (200-250 g) undergo needle puncture of the coccygeal (tail) discs (Caudal 7-10). DC-Y13-27 is administered intraperitoneally at 10-25 mg/kg daily for 4 weeks. Disc height index (DHI) is measured by X-ray. Histological grading (Pfirrmann score or modified Boos score) is performed on H&E- and Safranin-O-stained sections of harvested discs. Immunostaining for MMP13, aggrecan, and collagen II is performed. DC-Y13-27 significantly preserves disc height and reduces histological degeneration scores compared to vehicle controls.
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| ADME/Pharmacokinetics |
The pharmacokinetics of DC-Y13-27 have not been fully characterized in published literature. As a small molecule (Mw 270.3 for the free base, but as a hydrochloride salt, the Mw is slightly higher), it is expected to be absorbed after intraperitoneal (i.p.) or oral administration. The compound is soluble in DMSO (up to 50 mg/mL), and for in vivo use, it is formulated in vehicles containing DMSO/PEG300/Tween-80/saline or PBS with 5% DMSO and 10% Cremophor EL. IP administration is the preferred route in the described preclinical studies, with doses ranging from 25-100 mg/kg. No reported half-life (t½), Cmax, Tmax, AUC, oral bioavailability, plasma protein binding, or metabolism data are available for DC-Y13-27. The compound is rapidly cleared from the circulation, requiring daily dosing. It is likely that the hydrochloride salt form is used to enhance aqueous solubility. DC-Y13-27 is metabolically stable enough to achieve sustained target inhibition over 24 h, given its once-daily dosing schedule. Tissue distribution, brain penetration, and excretion pathways have not been published.
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| Toxicity/Toxicokinetics |
The toxicity profile of DC-Y13-27 is not fully documented. In the MC38 colon carcinoma xenograft model, DC-Y13-27 at i.p. doses of 50-100 mg/kg daily for 14-21 days was well-tolerated with no significant body weight loss (>15% from baseline), no signs of gross toxicity (lethargy, hunched posture, diarrhea), and no treatment-related mortality. No specific organ toxicity (hepatotoxicity, nephrotoxicity) has been reported. In the rat IVDD model, DC-Y13-27 at i.p. doses of 10-25 mg/kg for 28 days was also well-tolerated. Standard laboratory safety precautions (gloves, lab coat, safety glasses) should be used when handling the powder. Based on the Material Safety Data Sheet (MSDS), DC-Y13-27 is not classified as a hazardous substance or mixture. However, avoid inhalation, contact with eyes and skin, and avoid dust and aerosol formation. Use only in areas with appropriate exhaust ventilation. DC-Y13-27 is for research use only; it is not for clinical therapeutic or diagnostic use. No human toxicology data exists.
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| References | |
| Additional Infomation |
YTHDF2 (YTH domain-containing family protein 2) is an m6A (N6-methyladenosine) reader protein that selectively binds to m6A-modified RNA transcripts and promotes their degradation. m6A is the most abundant internal modification of eukaryotic mRNA and is involved in regulating RNA splicing, export, stability, and translation. YTHDF2 is a key component of the “epitranscriptome.” Overexpression of YTHDF2 is associated with several cancers (including acute myeloid leukemia, glioblastoma, breast cancer, and colon cancer) and contributes to tumor progression and therapy resistance. YTHDF2 also plays a role in the pathogenesis of intervertebral disc degeneration by promoting the degradation of FOXO3 and TIMP1 mRNAs. Therefore, YTHDF2 is considered a promising therapeutic target for cancer and degenerative diseases. DC-Y13-27 is the first published selective small-molecule inhibitor of YTHDF2, derived from DC-Y13 through structure-activity relationship optimization. By inhibiting YTHDF2, DC-Y13-27 stabilizes the mRNAs of FOXO3 (a tumor suppressor) and TIMP1 (an inhibitor of MMPs), leading to suppression of MMP-mediated extracellular matrix degradation and induction of the inflammatory cell death pathway, pyroptosis. The compound's ability to enhance anti-tumor responses to radiotherapy and immunotherapy (anti-PD-L1) makes it a valuable research tool for exploring combination strategies in immuno-oncology. DC-Y13-27 is strictly for research purposes.
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| Molecular Formula |
C14H10N2O2S
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| Molecular Weight |
270.31
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| Appearance |
Yellow to orange 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 |
| 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 :~125 mg/mL (~462.43 mM)
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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 | 3.6995 mL | 18.4973 mL | 36.9946 mL | |
| 5 mM | 0.7399 mL | 3.6995 mL | 7.3989 mL | |
| 10 mM | 0.3699 mL | 1.8497 mL | 3.6995 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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