| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| 50mg |
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| 100mg |
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| 250mg | |||
| Other Sizes |
| Targets |
Smoothened (Smo) antagonist (IC₅₀ = 10 nM in Shh-light2 cells; IC₅₀ = 10 nM in C3H10T1/2 alkaline phosphatase assay; IC₅₀ = 3–66 nM in rat cerebellar GCP proliferation assay; IC₅₀ = 4.6 nM for BC binding to human Smo; IC₅₀ = 14 nM for BC binding to mouse Smo)[1]
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|---|---|
| ln Vitro |
MRT-83 demonstrates full antagonist capabilities, reducing ShhN (3 nM)-induced rat GCP growth with IC50 (~3 nM). MRT-83 also suppresses SAG (0.01 μM)-induced GCP growth (IC50 ~6 nM). MRT-83 prevents BC binding to HEK-hSmo cells in a dose-dependent manner with an IC50 of 4.6 nM. MRT-83 inhibits BC binding to cells expressing mouse Smo with an IC50 of 14 nM, which matches well with its IC50 in Shh-light2 and alkaline phosphatase assays [1].
MRT-83 inhibits Gli-dependent luciferase activity in Shh-light2 cells stimulated with ShhN (5 nM) with an IC₅₀ of 10 nM. MRT-83 inhibits SAG-induced differentiation of C3H10T1/2 cells into alkaline phosphatase-positive osteoblasts with an IC₅₀ of 10 nM. MRT-83 inhibits ShhN- and SAG-induced proliferation of rat cerebellar granule cell precursors with IC₅₀ values of 3 nM and 6–66 nM, respectively. MRT-83 blocks Bodipycyclopamine binding to human and mouse Smo-expressing cells with IC₅₀ values of 4.6 nM and 14 nM, respectively. MRT-83 does not affect Wnt signaling in HEK293 cells transfected with a Tcf/Lef-dependent luciferase reporter.[1] |
| ln Vivo |
The health of the animals treated with ShhN in the presence of MRT-83 was comparable to that of the other groups; however, there was no longer any evidence of Ptc transcription being upregulated in the SVZ of these animals, which is consistent with complete inhibition of ShhN-mediated effects (8.7 ±2.4 Ptc+ cells/section, n=9), which were not different from effects mediated by the vehicle. In vivo, ShhN-induced Ptc transcriptional upregulation in the LV SVZ can be inhibited by MRT-83 but not by MRT-36 [1].
Stereotaxic injection of MRT-83 (200 ng) into the lateral ventricle of adult mice abolishes ShhN-induced up-regulation of Patched transcription in the subventricular zone, while an inactive analog MRT-36 does not.[1] |
| Cell Assay |
Gli-dependent luciferase reporter assay: Shh-light2 cells are incubated with ShhN (5 nM) and compounds for 40 hours, followed by luciferase activity measurement.
Alkaline phosphatase assay: C3H10T1/2 cells are incubated with SAG (0.1 μM) and compounds for 6 days, followed by alkaline phosphatase activity measurement. Cerebellar granule cell precursor proliferation assay: Primary rat cerebellar granule cell precursors are cultured with ShhN or SAG and compounds, and proliferation is assessed by [³H]thymidine incorporation. BC binding assay: HEK293 cells expressing human or mouse Smo are incubated with Bodipycyclopamine (5 nM) and compounds for 2 hours, fixed, and fluorescence is quantified. Ciliary Smo accumulation assay: C3H10T1/2 or NT2 cells are serum-starved and treated with SAG and compounds for 18 hours, followed by immunofluorescence staining for Smo and acetylated tubulin.[1] |
| Animal Protocol |
Adult mice are stereotaxically injected into the lateral ventricle with 5 μl of a solution containing ShhN (0.9 μg) alone or with MRT-83 (200 ng) or MRT-36 (110 ng) in 45% 2-hydroxypropyl-β-cyclodextrin/PBS. Animals are sacrificed 48 hours post-injection, and brains are processed for in situ hybridization.[1]
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| References | |
| Additional Infomation |
MRT-83 is an acylguanidine derivative with a structure conforming to the pharmacophore model of Smo antagonists, possessing three hydrogen-bonded acceptor groups and three hydrophobic regions. It blocks the transport of Smo to primary cilia in C3H10T1/2 and NT2 cells, similar to the effects of SANT-1-like inhibitors. It does not affect the Wnt signaling pathway, indicating its selectivity for the Hedgehog pathway. In multiple assays, it was more potent than cyclopamine and showed activity in vivo by inhibiting Shh-induced patched transcription in the mouse brain. [1]
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| Molecular Formula |
C31H30N4O5
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|---|---|
| Molecular Weight |
538.59
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| Exact Mass |
538.221
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| CAS # |
1263131-92-5
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| Related CAS # |
MRT-83 hydrochloride;1359944-60-7
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| PubChem CID |
136210511
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| Appearance |
White to light yellow solid powder
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| Density |
1.2±0.1 g/cm3
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| Index of Refraction |
1.608
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| LogP |
5.23
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
40
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| Complexity |
841
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
BKTMNLKJWHVZRN-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C31H30N4O5/c1-19-10-15-24(33-31(32)35-30(37)23-16-26(38-2)28(40-4)27(17-23)39-3)18-25(19)34-29(36)22-13-11-21(12-14-22)20-8-6-5-7-9-20/h5-18H,1-4H3,(H,34,36)(H3,32,33,35,37)
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| Chemical Name |
3,4,5-trimethoxy-N-[N'-[4-methyl-3-[(4-phenylbenzoyl)amino]phenyl]carbamimidoyl]benzamide
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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 : ~250 mg/mL (~464.17 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (3.86 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 20.8 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: ≥ 2.08 mg/mL (3.86 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 20.8 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 | 1.8567 mL | 9.2835 mL | 18.5670 mL | |
| 5 mM | 0.3713 mL | 1.8567 mL | 3.7134 mL | |
| 10 mM | 0.1857 mL | 0.9284 mL | 1.8567 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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