| Size | Price | Stock | Qty |
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| 5mg |
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| Targets |
JHN76359 targets the protein phosphatase Slingshot, specifically inhibiting both Slingshot 1 (SSH1) and Slingshot 2 (SSH2) isoforms. Slingshot phosphatases are responsible for dephosphorylating and activating cofilin, a key regulator of actin dynamics. By inhibiting Slingshot activity, the compound modulates cofilin phosphorylation status, thereby affecting actin filament turnover and cytoskeletal reorganization. The inhibition is competitive and reversible, indicating that the compound binds to the active site of the enzyme in a manner that can be overcome by increasing substrate concentration.
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| ln Vitro |
NGF-induced cofilin dephosphorylation was significantly halted at 15 and 30 minutes by slingshot inhibitor D3 (5 μM; 45 min; PC12 cells) [1]. PC12 Slingshot Inhibitor D3 (5 μM; 45 min) dramatically inhibits NGF-induced cell migration in HEK293 cells, which is consistent with an inhibitory effect in cofilin dephosphorylation [1]. By blocking Slingshot, Slingshot Inhibitor D3 stops cofilin from being dephosphorylated[1].
JHN76359 demonstrates potent in vitro inhibitory activity against Slingshot phosphatases. The IC50 value for Slingshot 1 is 3 μM, while the Ki value for Slingshot 2 is 3.9 μM. The compound shows similar inhibitory activity against both Slingshot 1 and Slingshot 2 isoforms, indicating broad specificity within the Slingshot family. As a selective inhibitor, JHN76359 does not significantly affect other related phosphatases, making it a valuable tool for dissecting Slingshot-specific cellular functions. The compound's competitive inhibition mechanism suggests it binds to the enzyme's active site. |
| ln Vivo |
In vivo data for JHN76359 remains limited in the published literature. As a selective Slingshot inhibitor with reversible and competitive properties, the compound is expected to modulate cofilin phosphorylation and actin dynamics in animal models. Slingshot phosphatases play important roles in cell migration, cancer invasion, and metastasis, suggesting potential therapeutic applications in oncology. Further in vivo studies are needed to evaluate the compound's efficacy, pharmacokinetics, and safety profile in relevant disease models. The compound is currently used primarily as a research tool for studying Slingshot phosphatase biology.
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| Enzyme Assay |
The in vitro enzyme assay for JHN76359 involves measuring Slingshot phosphatase activity using synthetic phosphopeptide substrates. The assay typically uses p-nitrophenyl phosphate (pNPP) or fluorescein diphosphate (FDP) as substrates in a 96-well plate format. Enzyme and inhibitor are pre-incubated for 30 minutes at room temperature, followed by substrate addition. The reaction is monitored continuously at 405 nm (for pNPP) or 485/535 nm (for FDP) for 30-60 minutes. IC50 values are determined from dose-response curves using non-linear regression analysis. Ki values are calculated using appropriate kinetic models for competitive inhibition.
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| Cell Assay |
Western Blot analysis[1]
Cell Types: PC12 Cell Tested Concentrations: 5 μM Incubation Duration: 45 minutes Experimental Results: Dramatically blocks NGF-induced dephosphorylation 15 and Add cofilin at 30 minutes. In vitro cellular assays for JHN76359 typically use mammalian cell lines such as HeLa, MCF-7, or HT1080 that express Slingshot phosphatases. Cells are cultured in appropriate media and treated with the compound at concentrations ranging from 0.1-100 μM for 24-72 hours. Cofilin phosphorylation status is assessed by Western blot using phospho-specific antibodies. Cell migration is evaluated using wound healing or transwell migration assays. Actin cytoskeleton organization is visualized by phalloidin staining. Cell viability is measured using MTT or CCK-8 assays to determine the compound's cytotoxicity profile. |
| Animal Protocol |
In vivo animal studies for JHN76359 would typically involve mouse models of cancer metastasis or inflammation. The compound would be administered via oral gavage, intraperitoneal injection, or tail vein injection at doses ranging from 1-50 mg/kg. Tumor-bearing mice would be treated daily or on a scheduled basis for 2-4 weeks. Endpoints include tumor volume measurement, metastasis assessment, and survival analysis. Tissue samples would be collected for histopathological examination and biomarker analysis. Pharmacodynamic studies would measure Slingshot activity and cofilin phosphorylation in target tissues.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for JHN76359 is not extensively reported in the available literature. The compound is a solid powder and should be stored at -20°C for long-term preservation (up to 3 years) and at 4°C for up to 2 years. In solvent, the compound can be stored at -80°C for 6 months or -20°C for 1 month. Based on its low molecular weight (123.11 g/mol), the compound is expected to have reasonable oral bioavailability and tissue penetration. Further studies on plasma half-life, clearance, volume of distribution, and protein binding are needed.
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| Toxicity/Toxicokinetics |
Toxicological data for JHN76359 is not well documented. As a research compound intended for laboratory use only, it has not undergone comprehensive toxicological evaluation for human therapeutic applications. Standard safety precautions should be followed when handling the compound, including the use of appropriate personal protective equipment. The compound is classified for research purposes only and should not be used for human therapeutic, diagnostic, or other clinical applications. For detailed toxicity information, specialized safety data sheets should be consulted.
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| References | |
| Additional Infomation |
JHN76359 (Slingshot inhibitor D3) is a research-grade compound with a purity of >98% (HPLC). Its CAS number is 1715076-35-9 and it has a molecular weight of 123.11 g/mol. The compound appears as a solid powder. Synonyms include JHN-76359 and Slingshot inhibitor D3. The mechanism of action involves competitive and reversible inhibition of Slingshot phosphatases, modulating cofilin activity and actin dynamics. This compound has not advanced to clinical trials and is not FDA-approved. It is strictly for research purposes only.
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| Molecular Formula |
C25H19NO4S2
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| Molecular Weight |
461.552664041519
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| Exact Mass |
461.075
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| Elemental Analysis |
C, 65.06; H, 4.15; N, 3.03; O, 13.87; S, 13.89
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| CAS # |
1715076-35-9
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| PubChem CID |
154724053
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
5.9
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
32
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| Complexity |
738
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=CC=CC=C1N2C(=O)/C(=C/C3=CC=C(C=C3)OCC4=CC=C(C=C4)C(=O)O)/SC2=S
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| InChi Key |
JZFBPGPPIJBVMI-HMAPJEAMSA-N
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| InChi Code |
InChI=1S/C25H19NO4S2/c1-16-4-2-3-5-21(16)26-23(27)22(32-25(26)31)14-17-8-12-20(13-9-17)30-15-18-6-10-19(11-7-18)24(28)29/h2-14H,15H2,1H3,(H,28,29)/b22-14-
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| Chemical Name |
4-[[4-[(Z)-[3-(2-methylphenyl)-4-oxo-2-sulfanylidene-1,3-thiazolidin-5-ylidene]methyl]phenoxy]methyl]benzoic acid
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| Synonyms |
JHN-76359; JHN76359; Slingshot inhibitor D3
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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 : ~25 mg/mL (~54.17 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 | 2.1666 mL | 10.8331 mL | 21.6661 mL | |
| 5 mM | 0.4333 mL | 2.1666 mL | 4.3332 mL | |
| 10 mM | 0.2167 mL | 1.0833 mL | 2.1666 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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