| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| Other Sizes |
| Targets |
HhAntag targets the Hedgehog signaling pathway by inhibiting the Smoothened (SMO) receptor. SMO is a G protein-coupled receptor that is a key transducer in the Hh pathway. By blocking SMO, HhAntag prevents the activation of downstream signaling, including the inhibition of GLI1-mediated transcription. This disruption of the Hh pathway leads to antitumor effects.
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| ln Vitro |
HhAntag (2-30 µM; 72 hours) inhibits Hh signaling pathway-sensitive cells with IC50 values ranging from 2 µM to >30 µM[1]. It is ten times more powerful than the natural product SMO antagonist cyclopamine in suppressing Hh pathway activity. With IC50 values of 30 µM, 5.4 µM, and 5.8 µM, HhAntag inhibits AsPC-1, BXPC-3, CFPAC, HPAC, HPAF-II, KP4, Panc 03.27, PA-TU-8902, PSN-1, and SU.86.86 cells. These are 2.7 µM, 6.2 µM, 10.3 µM, 2.5 µM, 2.9 µM, 5.8 µM, and 2.7 µM[1]. A Hh-responsive human mesenchymal cell line (HEPM) expressing GLI luciferase reporter construct (HEPM-rep) requires 100 nM of HhAntag to completely inhibit Hh signaling; this is greater than the value required for inhibition, which is 400 times lower, and in the most sensitive cancer cell line (1.9 μM), cell growth increases by 50% [1].
HhAntag inhibits Shh signaling in a reporter assay with an EC50 of 0.4 μM. It inhibits the proliferation of GNP cells when used at concentrations ranging from 0.1 nM to 25 μM. Its activity is confirmed in various cell-based assays measuring Hh pathway activity. |
| ln Vivo |
In HT55 and HT-29 colorectal cell line xenograft models, HhAntag (oral treatment; 75 mg/kg or 100 mg/kg; twice daily; 25 days) significantly slowed tumor growth, with an average tumor growth inhibition rate of 29% and 48%, respectively. DLD-1 xenograft development is unaffected by HhAntag [1].
HhAntag has shown antitumor activity in vivo. It has been evaluated in animal models of cancers driven by aberrant Hh signaling. Its ability to inhibit GLI1-mediated transcription and block the Hh pathway makes it a promising candidate for cancer therapy. However, specific in vivo data is limited. |
| Enzyme Assay |
The primary in vitro assay for HhAntag is a cell-based reporter assay measuring Shh signaling. Cells are transfected with a GLI-responsive luciferase reporter and treated with Shh. The compound's ability to inhibit Shh-induced luciferase activity is measured, and the EC50 of 0.4 μM is determined. Proliferation assays using GNP cells are also used to assess its activity.
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| Cell Assay |
Cellular assays for HhAntag involve treating cancer cell lines with activated Hh signaling with the compound. The inhibition of GLI1-mediated transcription is assessed by measuring the expression of Hh target genes, such as GLI1 and PTCH1, by qPCR. Cell proliferation and viability are measured using standard assays.
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| Animal Protocol |
Animal/Disease Models: Primary human xenografts (DLD-1, HT55 and HT-29 cells) from 6-8 week old female CD1 nu/nu (nude) mice [1]
Doses: 75 mg/kg or 100 mg/kg twice (two times) daily ; 25-day Experimental Results: Caused growth delay in HT55 and HT-29 xenografts, but had no effect on DLD-1 xenografts. In vivo efficacy of HhAntag has been evaluated in mouse models of Hh-driven cancers. The compound is administered, and its effects on tumor growth and survival are monitored. Pharmacodynamic markers, such as the inhibition of Hh target gene expression in tumor tissue, are also assessed. |
| ADME/Pharmacokinetics |
HhAntag has a molecular weight of 450.92 g/mol and a molecular formula of C24H23ClN4O3. Its IUPAC name is N-(4-Chloro-3-(6-(dimethylamino)-1H-benzo[d]imidazol-2-yl)phenyl)-3,5-dimethoxybenzamide. It is soluble in DMSO. It is typically stored at 2-8°C.
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| Toxicity/Toxicokinetics |
HhAntag is a research compound that is not intended for human use. Its toxicological profile is not extensively characterized in the available literature. As with all research chemicals, standard safety precautions should be followed when handling.
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| References | |
| Additional Infomation |
HhAntag is a small-molecule inhibitor of the Hedgehog signaling pathway that targets the Smoothened (SMO) receptor. It inhibits GLI1-mediated transcription and has antitumor activity. It is used in cancer research to study the Hh pathway and is available from various chemical suppliers for research purposes.
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| Molecular Formula |
C24H23CLN4O3
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|---|---|
| Molecular Weight |
450.92
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| Exact Mass |
450.146
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| CAS # |
496794-70-8
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| PubChem CID |
9911630
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
5.602
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
32
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| Complexity |
626
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
UBHJFPVTEATUFS-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C24H23ClN4O3/c1-29(2)16-6-8-21-22(12-16)28-23(27-21)19-11-15(5-7-20(19)25)26-24(30)14-9-17(31-3)13-18(10-14)32-4/h5-13H,1-4H3,(H,26,30)(H,27,28)
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| Chemical Name |
N-[4-chloro-3-[6-(dimethylamino)-1H-benzimidazol-2-yl]phenyl]-3,5-dimethoxybenzamide
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| Synonyms |
Hh Antag Hh-Antag HhAntag
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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 (~221.77 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 3 mg/mL (6.65 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 30.0 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: ≥ 3 mg/mL (6.65 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 30.0 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 | 2.2177 mL | 11.0884 mL | 22.1769 mL | |
| 5 mM | 0.4435 mL | 2.2177 mL | 4.4354 mL | |
| 10 mM | 0.2218 mL | 1.1088 mL | 2.2177 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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