| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg | |||
| Other Sizes |
| Targets |
The primary target of Robotnikinin is the Sonic Hedgehog (Shh) ligand. Robotnikinin binds directly to Shh with a Kd value of 3.1 μM. By binding to the Shh ligand, Robotnikinin inhibits Shh signaling upstream of Smo. The Hedgehog signaling pathway plays a critical role in embryonic development, cell proliferation, and tissue patterning. Dysregulation of Hedgehog signaling is associated with various cancers and developmental disorders. Robotnikinin's direct binding to the Shh ligand provides a unique tool for studying the role of Shh in Hedgehog signaling.
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| ln Vitro |
Robotnikinin exhibits ShhN-binding ability at doses ranging from 1.56 μM to 25 μM. Based on kinetic data, a KD of 3.1 μM was determined[2]. Robotnikinin (50 µM) inhibits the rise in LC3-II caused by ShhN[3]. In H929 and U266 cell lines cocultured with HS-5 cells, robotnikinin (5 μM) downregulates classical NF-κB pathway proteins, indicating a connection between Hh signaling and the NF-κB pathway in MM[4].
In vitro, Robotnikinin demonstrates ShhN-binding ability at doses ranging from 1.56 μM to 25 μM. Based on kinetic data, a Kd of 3.1 μM was determined. Robotnikinin (50 µM) inhibits the rise in LC3-II caused by ShhN. In H929 and U266 cell lines cocultured with HS-5 cells, Robotnikinin (5 μM) downregulates classical NF-κB pathway proteins, indicating a connection between Hh signaling and the NF-κB pathway in multiple myeloma. The compound's ability to inhibit Shh signaling can be evaluated in various cell-based assays measuring Hedgehog pathway activity. |
| ln Vivo |
In vivo, Robotnikinin has been evaluated in models of Hedgehog-dependent tumors. The compound's ability to inhibit Shh signaling upstream of Smo makes it a valuable tool for studying the role of Hedgehog signaling in tumor growth and progression. By modulating this pathway, Robotnikinin can be used to study Hedgehog-mediated regulation of cell proliferation, differentiation, and tissue patterning. In skin and cosmetic research, it is used to study Hedgehog-mediated regulation of cell proliferation and differentiation. Further in vivo studies are needed to fully characterize its efficacy and safety in animal models.
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| Enzyme Assay |
In vitro enzyme/receptor binding studies for Robotnikinin focus on its interaction with the Shh ligand. Binding affinity to Shh can be assessed using surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC). A Kd value of 3.1 μM has been determined from kinetic data. The compound's ability to inhibit Shh signaling can be evaluated using Gli-luciferase reporter assays in cells expressing the Hedgehog pathway. Selectivity profiling against other ligands and signaling pathways can be performed to assess specificity. These methods are for research purposes only.
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| Cell Assay |
RT-PCR[4]
Cell Types: NCI -H929 or U266 cells. Tested Concentrations: 5 μM. Incubation Duration: 48 h. Experimental Results: Downregulated classical NF-κB pathway proteins in H929 and U266 cell lines cocultured with HS-5 cells. In vitro cell-based assays for Robotnikinin evaluate its effects on Hedgehog signaling. Cells expressing the Hedgehog pathway (e.g., H929 and U266 multiple myeloma cell lines) are treated with Robotnikinin at various concentrations (typically 0.1-50 µM). Hedgehog pathway activity is measured using Gli-luciferase reporter assays or by measuring the expression of Hedgehog target genes (e.g., GLI1, PTCH1) by qRT-PCR. NF-κB pathway proteins are measured by Western blot. Cell proliferation is assessed using MTT or CellTiter-Glo assays. RT-PCR can be performed to measure gene expression changes. Standard cell culture conditions are used with appropriate media and supplementation. |
| Animal Protocol |
In vivo animal studies for Robotnikinin would typically involve mouse models of Hedgehog-dependent tumors or developmental disorders. Tumor-bearing mice are treated with Robotnikinin via intraperitoneal or oral administration at various doses. Tumor volumes are measured regularly using calipers, and body weight is monitored for toxicity assessment. Pharmacodynamic studies evaluate Hedgehog pathway inhibition by measuring Gli1 expression in tumor tissues. The compound's ability to inhibit Shh signaling upstream of Smo can be distinguished from Smo inhibitors in these models. All procedures must comply with institutional animal care guidelines.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of Robotnikinin are not fully characterized in publicly available literature. The compound has a molecular weight of 454.95 g/mol. It is soluble in DMSO and should be stored at -20°C. The compound's half-life, Cmax, AUC, and clearance would need to be determined in preclinical pharmacokinetic studies. Information concerning product stability, particularly in solution, has rarely been reported. The compound is for research use only and is not approved for clinical use.
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| Toxicity/Toxicokinetics |
The toxicity profile of Robotnikinin is not fully characterized in publicly available literature. The compound is classified for research use only and not for human consumption. Standard safety precautions for handling chemical compounds apply, including the use of personal protective equipment and working in a chemical fume hood. Preclinical toxicology studies would be required for clinical development. The compound should be handled with care due to its biological activity.
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| References | |
| Additional Infomation |
It binds to the sound hedgehog factor protein, blocking its signaling pathway; the structure is described in the first article.
Additional information: Robotnikinin has the CAS number 1132653-79-2 and the molecular formula C₂₅H₂₇ClN₂O₄. The compound is a first-in-class small molecule inhibitor of Shh signaling that acts upstream of Smo, binding directly to Shh with a Kd value of 3.1 μM. Robotnikinin has a purity of >98% and a molecular weight of 454.95 g/mol. The compound is also known as N-[(4-chlorophenyl)methyl]-2-[(2R,6S,8E)-5,12-dioxo-2-phenyl-1-oxa-4-azacyclododec-8-en-6-yl]acetamide. Robotnikinin is used in research to study Hedgehog-mediated regulation of cell proliferation, differentiation, and tissue patterning. This product is for research use only and is not approved for clinical or therapeutic applications. |
| Molecular Formula |
C25H27CLN2O4
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| Molecular Weight |
454.9459
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| Exact Mass |
454.166
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| CAS # |
1132653-79-2
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| PubChem CID |
25109985
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| Appearance |
White to off-white solid powder
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| LogP |
4.823
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
32
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| Complexity |
654
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| Defined Atom Stereocenter Count |
2
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| SMILES |
C\1CC(=O)O[C@@H](CNC(=O)[C@@H](C/C=C1)CC(=O)NCC2=CC=C(C=C2)Cl)C3=CC=CC=C3
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| InChi Key |
DRDSZZCLAHXSAE-BQIDRLATSA-N
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| InChi Code |
InChI=1S/C25H27ClN2O4/c26-21-13-11-18(12-14-21)16-27-23(29)15-20-9-5-2-6-10-24(30)32-22(17-28-25(20)31)19-7-3-1-4-8-19/h1-5,7-8,11-14,20,22H,6,9-10,15-17H2,(H,27,29)(H,28,31)/b5-2+/t20-,22-/m0/s1
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| Chemical Name |
N-[(4-chlorophenyl)methyl]-2-[(2R,6S,8E)-5,12-dioxo-2-phenyl-1-oxa-4-azacyclododec-8-en-6-yl]acetamide
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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 | 2.1980 mL | 10.9902 mL | 21.9804 mL | |
| 5 mM | 0.4396 mL | 2.1980 mL | 4.3961 mL | |
| 10 mM | 0.2198 mL | 1.0990 mL | 2.1980 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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