| Targets |
Brachyury (TBXT) - a transcription factor and oncogenic driver in chordoma.
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| ln Vitro |
DHC-156 downmodulates brachyury with a DC50 value of 4.1 microM and achieves greater than 99% downmodulation at 10 microM. It eliminates kinase-inhibition, sparing all wild-type kinases tested, and induces irreversible impairment of chordoma tumor cell growth. DHC-156 downmodulates brachyury post-translationally in a proteasome- and lysosome-independent manner.
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| ln Vivo |
Currently, no in vivo data are available for DHC-156. Its reported ability to irreversibly impair chordoma tumor cell growth suggests potential for future in vivo validation, but as of now, no animal studies or efficacy data in living systems have been published.
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| Enzyme Assay |
Not reported. A generic assay would involve immobilizing recombinant brachyury protein or brachyury-binding peptides on a sensor chip for surface plasmon resonance (SPR). Increasing concentrations of DHC-156 would be flowed over the chip, and binding affinity (KD) would be calculated from sensorgram association/dissociation rates.
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| Cell Assay |
Cell line: Chordoma cell lines (e.g., U-CH1, MUG-Chor1) cultured in IMDM + 10% FBS + 1% penicillin/streptomycin. Treatment: cells treated with a dose range of DHC-156 (0.1-50 microM) for 24-72 h. Brachyury level quantification: Western blot or ELISA. Cell growth: CCK-8 assay or direct cell counting. Apoptosis: Annexin V/PI staining followed by flow cytometry.
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| Animal Protocol |
No published in vivo data were found. A general protocol would involve establishing subcutaneous chordoma xenografts in nude mice (e.g., 5×10⁶ U-CH1 cells flank injection). When tumors reach ~100 mm3, mice are randomized to receive DHC-156 (e.g., 30 mg/kg) or vehicle by oral gavage daily for 3-4 weeks. Tumor volume is measured twice weekly, and tumors are harvested for brachyury IHC.
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| ADME/Pharmacokinetics |
No data were found. Since DHC-156 is derived from afatinib (an oral TKI, F~40% in humans), typical PK parameters for a related compound would involve plasma protein binding, clearance, volume of distribution, and oral bioavailability. No specific studies were reported.
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| Toxicity/Toxicokinetics |
No formal toxicity studies were found. Given its derivation from a clinically approved drug (afatinib), DHC-156 may share some safety liabilities, though the optimization eliminated off-target kinase inhibition. No specific toxicology data were reported.
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| References | |
| Additional Infomation |
DHC-156 is a research-grade covalent small molecule developed to directly target brachyury. The feasibility of direct brachyury modulation is demonstrated, and DHC-156 may be further developed into more potent tool compounds and therapies. It was developed through structure-based drug design, aided by mass spectrometry and X-ray crystallography. Available in purities ≥98%, soluble in DMSO to 100 mM.
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| Appearance |
White to light yellow 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 : ~100 mg/mL (~203.06 mM; with ultrasonication)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.08 mM)(saturation unknown) in 10% DMSO 40% PEG300 5% Tween-80 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 and add it to 400 μL PEG300, mix well; then add 50 μL Tween-80 to the above system, mix well; then continue to add 450 μL of normal saline to make up to 1 mL. Preparation of normal saline: Dissolve 0.9 g of sodium chloride in ddH₂O and make up to 100 mL to obtain a clear and transparent normal saline solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.08 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 and add it to 900 μL corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
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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