| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 500mg | |||
| Other Sizes |
| Targets |
Clathrin-IN-1 selectively inhibits amphiphysin association of the clathrin terminal domain (TD) with an IC₅₀ value of 12 μM. By binding to the clathrin terminal domain, it disrupts the function of clathrin in endocytosis, interfering with receptor-mediated endocytosis, entry of HIV, and synaptic vesicle recycling.
|
|---|---|
| ln Vitro |
HeLa cells treated with Clathrin-IN-1 (Pitstops 2) prior to incubation exhibit a dose-dependent reduction in Tf uptake, with an IC50 value of 12–5 μM. When 30 μM Clathrin-IN-1 was applied, Tf endocytosis was totally inhibited. After one to three hours of drug washout, the HeLa cells' clathrin-IN-1-induced inhibition of Tf endocytosis was fully restored. The Tf uptake IC50 in U2OS cells is 9.7 μM. Additionally, Pitstop 2 strongly inhibits the absorption of EGF[1]. In HeLa cells, clathrin-IN-1 (Pitstops 2) effectively and selectively decreased HIV-1 infectivity by more than 90%[1]. Pitstop-induced clathrin TD function suppression abruptly disrupts HIV entry, receptor-mediated endocytosis, and synaptic vesicle recycling. The durations of clathrin coat components, such as FCHo, clathrin, and dynamin, dramatically increase during endocytosis inhibition, indicating that clathrin TD governs coated pit dynamics[1].
In vitro, Clathrin-IN-1 potently and specifically reduced HIV-1 infectivity by >90% in HeLa cells. Preincubation of HeLa cells with Clathrin-IN-1 leads to a dose-dependent inhibition of transferrin (Tf) uptake with an IC₅₀ value of 12-5 μM, and 30 μM completely blocks Tf endocytosis. This block is completely reversed within 1-3 hours of drug washout. In U2OS cells, the IC₅₀ for Tf uptake is 9.7 μM. |
| ln Vivo |
Clathrin-IN-1 trapped HeLa cells in metaphase through loss of mitotic spindle integrity and activation of the spindle assembly checkpoint, phenocopying clathrin depletion and aurora A kinase inhibition. This suggests that clathrin function is essential for mitotic spindle stability and proper cell division.
|
| Enzyme Assay |
For non-cellular in vitro enzyme/receptor binding assays, Clathrin-IN-1 is evaluated for its binding affinity to the clathrin terminal domain. Purified clathrin terminal domain protein is used in binding assays such as surface plasmon resonance (SPR) or fluorescence polarization to determine the compound's binding affinity and to measure the IC₅₀ for inhibition of amphiphysin association.
|
| Cell Assay |
For in vitro cellular assays, Clathrin-IN-1 is tested in cell lines such as HeLa or U2OS. Cells are preincubated with the compound at various concentrations, and clathrin-mediated endocytosis is assessed by measuring the uptake of fluorescently labeled transferrin (Tf) or epidermal growth factor (EGF). IC₅₀ values are determined from dose-response curves. HIV-1 infectivity assays may also be performed.
|
| Animal Protocol |
For in vivo animal studies, Clathrin-IN-1 has potential for anti-cancer research, but specific in vivo protocols are not extensively documented. Based on its mechanism, potential studies could involve administration to tumor-bearing mouse models to assess effects on tumor growth, metastasis, and survival.
|
| ADME/Pharmacokinetics |
Pharmacokinetic data for Clathrin-IN-1 are limited. The compound is soluble in DMSO (34.29 mg/mL, 72.44 mM) and is typically formulated for in vitro studies. Further studies would be needed to determine its absorption, distribution, metabolism, and excretion profile for in vivo applications.
|
| Toxicity/Toxicokinetics |
Clathrin-IN-1 is generally considered to have low toxicity in vitro, as evidenced by the reversibility of its effects upon drug washout. Comprehensive toxicology studies would be required for therapeutic development. The compound has a molecular weight of 473.36 and a molecular formula of C₂₀H₁₃BrN₂O₃S₂.
|
| References | |
| Additional Infomation |
Clathrin-IN-1 is a selective clathrin-mediated endocytosis (CME) inhibitor with an IC₅₀ of 12 μM for inhibiting amphiphysin association with the clathrin terminal domain. It acutely interferes with receptor-mediated endocytosis, HIV entry, and synaptic vesicle recycling. It has potential for anti-cancer research due to its effects on mitotic spindle integrity and cell division.
|
| Molecular Formula |
C20H13BRN2O3S2
|
|---|---|
| Exact Mass |
471.955
|
| CAS # |
1332879-52-3
|
| Related CAS # |
Pitstop 2;1419320-73-2
|
| PubChem CID |
136246422
|
| Appearance |
Typically exists as solid at room temperature
|
| Density |
1.6±0.1 g/cm3
|
| Boiling Point |
652.3±65.0 °C at 760 mmHg
|
| Flash Point |
348.3±34.3 °C
|
| Vapour Pressure |
0.0±2.0 mmHg at 25°C
|
| Index of Refraction |
1.723
|
| LogP |
5.57
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
28
|
| Complexity |
767
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
O=C1N=C(NS(=O)(=O)C2C=CC=C3C=CC=CC=23)S/C/1=C\C1C=CC(Br)=CC=1
|
| InChi Key |
CGDLWHGPJPVPDU-ATVHPVEESA-N
|
| InChi Code |
InChI=1S/C20H13BrN2O3S2/c21-15-10-8-13(9-11-15)12-17-19(24)22-20(27-17)23-28(25,26)18-7-3-5-14-4-1-2-6-16(14)18/h1-12H,(H,22,23,24)/b17-12-
|
| Chemical Name |
(NZ)-N-[(5Z)-5-[(4-bromophenyl)methylidene]-4-oxo-1,3-thiazolidin-2-ylidene]naphthalene-1-sulfonamide
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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.) |
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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
