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20-HEDE (WIT 002) is an antagonist of 20-hydroxyeicosatetraenoic acid (20-HETE).

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

20-HEDE Chemical Structure

20-HEDE Chemical Structure

CAS No. : 240427-90-1

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Customer Review

Based on 2 publication(s) in Google Scholar

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  • Biological Activity

  • Protocol

  • Purity & Documentation

  • References

  • Customer Review

Description

20-HEDE (WIT 002) is an antagonist of 20-hydroxyeicosatetraenoic acid (20-HETE).

IC50 & Target

20-HEDE (WIT 002)[1].

In Vitro

ω-hydroxylation activity toward arachidonic acid is high in A549 cells, thus, A549 cells are treated with HET0016 or WIT 002 in the invasion assays, and both of them significantly decrease invasion[2]. WIT 002 inhibits proliferation of 786-O and 769-P renal adenocarcinoma cells, but HET0016 and WIT 002 fail to inhibit proliferation of normal renal epithelial cells RPTC[2][3].

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

In Vivo

The effect of the 20-HETE antagonist, WIT 002 on the growth of 786-O clear cell renal carcinoma is assessed in ectopic mouse model of renal tumor. The growth of tumors is significantly suppressed by WIT 002 administered daily to athymic nude mice implanted subcutaneously with cells 786-O. Tumor growth is inhibited by 84%±128%. It is of note that in these experiments WIT 002 treatment start only after the tumor is seeded for 7-14 days and is relatively large 0.1 cm. Thus, WIT 002 is effective at arresting the growth of a fairly advanced tumor[3].

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

Molecular Weight

324.50

Formula

C20H36O3

CAS No.
Appearance

Liquid (Density: 0.959±0.06 g/cm3)

Color

Off-white to yellow

SMILES

O=C(O)CCCC/C=C\CCCCCCC/C=C\CCCCO

Shipping

Room temperature in continental US; may vary elsewhere.

Storage
Pure form -20°C 3 years
In solvent -80°C 6 months
-20°C 1 month
Solvent & Solubility
In Vitro: 

DMSO : ≥ 180 mg/mL (554.70 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

*"≥" means soluble, but saturation unknown.

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 3.0817 mL 15.4083 mL 30.8166 mL
5 mM 0.6163 mL 3.0817 mL 6.1633 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, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

  • Molarity Calculator

  • Dilution Calculator

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

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Volume
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Molecular Weight *

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

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

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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    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 4.5 mg/mL (13.87 mM); Clear solution

    This protocol yields a clear solution of ≥ 4.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (45.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
  • Protocol 2

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

    Solubility: ≥ 4.5 mg/mL (13.87 mM); Clear solution

    This protocol yields a clear solution of ≥ 4.5 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 (45.0 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: ≥98.0%

References
Cell Assay
[2][3]

Human NSCLC cell lines (e.g A549) are seeded onto the upper well of the chamber. Subsequently, serum-free medium with HET0016 (10 μM), WIT002 (10 μM), WIT003 (0.01, 0.1, 1 μM), or ethanol as a control is added to the upper chamber, while the lower well is filled to the top (500 μL) with RPMI-1640 containing 5% fetal calf serum (FCS) as a chemoattractant. Cells are allowed to migrate for 5 h. Cells that have invaded to the bottom surface of the filter are counted with an ocular micrometer in a blinded manner, counting a minimum of 10 high-powered fields (HPF) [2]. Human renal cell adenocarcinoma lines (RPTC) 786-O or 769-P cells are plated and next day (0 hrs) transferred to serum-free medium containing either EGF or ET-1. Cells are exposed either to 10 μM WIT002 or vehicle. Cell counting is performed at the day of transfer to serum free medium (0 hrs) and 24, 48 and 72 hrs thereafter. Medium is changed to fresh containing mitogens and drugs every 24 hrs. Data presented are characteristic experiment from at least two separate experiments, each performed in triplicate[3].

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

Animal Administration
[3]

Mice[3]
Experiments are carried out on 6-week immunodefficient athymic nude mice weighing 20-26 g. Animals are acclimated for 1 week prior to injection with renal cell carcinoma. Immediately before each implantation, the cells are trypsinized, counted and resuspended in 10% serum containing RPMI media. The concentration of cells is adjusted to 40 millions/mL and 4 million cells/animal are injected subcutaneously. The cells are allowed to grow for 7-15 days until the size of the tumors reach approximately 0.1 cm3. The mice then receive daily subcutaneous injection (s.c.) injections of WIT 002 (10 mg/kg/day in 200 μL) in an isotonic NaPO4 buffer (pH 9.0) or vehicle (0.1 M NaPO4 solution pH 9.0). The diameter of the tumor is measured on every 3-4 days for 2 weeks using precision calibers. At the end of the experiment the mice are euthanized with CO2 and the tumors were excised to confirm the diameter measurements[3].

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, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 3.0817 mL 15.4083 mL 30.8166 mL 77.0416 mL
5 mM 0.6163 mL 3.0817 mL 6.1633 mL 15.4083 mL
10 mM 0.3082 mL 1.5408 mL 3.0817 mL 7.7042 mL
15 mM 0.2054 mL 1.0272 mL 2.0544 mL 5.1361 mL
20 mM 0.1541 mL 0.7704 mL 1.5408 mL 3.8521 mL
25 mM 0.1233 mL 0.6163 mL 1.2327 mL 3.0817 mL
30 mM 0.1027 mL 0.5136 mL 1.0272 mL 2.5681 mL
40 mM 0.0770 mL 0.3852 mL 0.7704 mL 1.9260 mL
50 mM 0.0616 mL 0.3082 mL 0.6163 mL 1.5408 mL
60 mM 0.0514 mL 0.2568 mL 0.5136 mL 1.2840 mL
80 mM 0.0385 mL 0.1926 mL 0.3852 mL 0.9630 mL
100 mM 0.0308 mL 0.1541 mL 0.3082 mL 0.7704 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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20-HEDE
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HY-101527
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