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  3. ZCL278

ZCL278 is a selective Cdc42 modulator that directly binds to Cdc42 and inhibits its functions with Kd of 11.4 μM for Cdc42-ZCL278 affinity in surface plasmon resonance (SPR) experiment.

For research use only. We do not sell to patients.

ZCL278 Chemical Structure

ZCL278 Chemical Structure

CAS No. : 587841-73-4

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10 mM * 1 mL in DMSO
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Based on 2 publication(s) in Google Scholar

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  • Protocol

  • Purity & Documentation

  • References

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Description

ZCL278 is a selective Cdc42 modulator that directly binds to Cdc42 and inhibits its functions with Kd of 11.4 μM for Cdc42-ZCL278 affinity in surface plasmon resonance (SPR) experiment.

IC50 & Target

Kd: 11.4 μM (Cdc42)[1]

In Vitro

ZCL278 as a potent, cell-permeable Cdc42-specific inhibitor that suppresses actin-based cellular functions, including Golgi organization and cell motility.In Swiss 3T3 fibroblast cultures, ZCL278 abolishes microspike formation and disrupted GM130-docked Golgi structures, two of the most prominent Cdc42-mediated subcellular events. ZCL278 reduces the perinuclear accumulation of active Cdc42 in contrast to NSC23766, a selective Rac inhibitor. ZCL278 suppresses Cdc42-mediated neuronal branching and growth cone dynamics as well as actin-based motility and migration in a metastatic prostate cancer cell line (i.e., PC-3) without disrupting cell viability[1]. ZCL278 inhibits Cdc42 function as an entry inhibitor for Junin virus (JUNV) and for vesicular stomatitis virus, lymphocytic choriomeningitis virus, and dengue virus but not for the nonenveloped poliovirus. In cells, ZCL278 is shown to efficiently inhibit chemically induced filopodium formation, a process dependent on Cdc42 activity. Dose-response experiments are first carried out in Vero cells, and while ZCL278 is not toxic at concentrations up to 200 μM, ZCL278 inhibits JUNV with IC50 of ~14 μM, as measured by flow cytometry[2].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

In Vivo

ZCL278 reduces the JUNV RNA load in the spleen more than 33-fold, with JUNV RNA being undetectable in 5 out of 8 mice. These results are similar to those seen in Gabapentin-treated mice, demonstrating that ZCL278 can abrogate JUNV replication[2].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Molecular Weight

584.89

Formula

C21H19BrClN5O4S2

CAS No.
Appearance

Solid

Color

White to light yellow

SMILES

O=C(NC(NC1=CC=C(S(=O)(NC2=NC(C)=CC(C)=N2)=O)C=C1)=S)COC3=CC=C(Br)C=C3Cl

Shipping

Room temperature in continental US; may vary elsewhere.

Storage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 2 years
-20°C 1 year
Solvent & Solubility
In Vitro: 

DMSO : 50 mg/mL (85.49 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 1.7097 mL 8.5486 mL 17.0972 mL
5 mM 0.3419 mL 1.7097 mL 3.4194 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.5 mg/mL (4.27 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% Corn Oil

    Solubility: ≥ 2.5 mg/mL (4.27 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.

In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation

Purity: 98.46%

References
Kinase Assay
[1]

Lyophilized Cdc42 protein is reconstituted to 5 mg/mL in a buffer consisting of 50 mM Tris, 0.5 mM MgCl2, 50 mM NaCl, 3% (wt/vol) sucrose, and 0.6% dextran. The stock solution is then diluted to 1 μM in 5 mM phosphate buffer, pH 7.4. Into a quartz cuvettete containing Cdc42 solution, aliquots of ZCL278 are added and incubated for 5 min before each fluorescent measurement. The excitation wavelength is 275 nm, and the fluorescence of tryptophan at 350 nm is measured after each addition. The titration curve is fitted using the equimolar specific binding model in GraphPad, and the Kd is calculated[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Assay
[1]

To determine cell viability, PC-3 cells are incubated for 24 h with or without the Cdc42 activator, ZCL278, or NSC23766. By using the trypan blue dye exclusion method, the numbers of live and dead cells are obtained with a Countess Automated Cell Counter. P values are assigned in each experiment, and any null hypothesis with probability level <95% is rejected[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Administration
[2]

Mice[2]
Four-week-old C57BL/6 mice receive intravenous injections of Gabapentin or ZCL278 (100 μg/g, i.p.). At 1 h after treatment, the mice are inoculated intraperitoneally with JUNV Candid #1 (1×106 PFU) in no more than 1 mL with a 27 1/2-gauge needle. At the end of the experiment, the mice are sacrificed, their spleens are homogenized with a Dounce homogenizer and centrifuged to generate a cell pellet and supernatant, and RNA expression levels are determined.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 1.7097 mL 8.5486 mL 17.0972 mL 42.7431 mL
5 mM 0.3419 mL 1.7097 mL 3.4194 mL 8.5486 mL
10 mM 0.1710 mL 0.8549 mL 1.7097 mL 4.2743 mL
15 mM 0.1140 mL 0.5699 mL 1.1398 mL 2.8495 mL
20 mM 0.0855 mL 0.4274 mL 0.8549 mL 2.1372 mL
25 mM 0.0684 mL 0.3419 mL 0.6839 mL 1.7097 mL
30 mM 0.0570 mL 0.2850 mL 0.5699 mL 1.4248 mL
40 mM 0.0427 mL 0.2137 mL 0.4274 mL 1.0686 mL
50 mM 0.0342 mL 0.1710 mL 0.3419 mL 0.8549 mL
60 mM 0.0285 mL 0.1425 mL 0.2850 mL 0.7124 mL
80 mM 0.0214 mL 0.1069 mL 0.2137 mL 0.5343 mL
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Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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