2. Metabolic Enzyme/Protease Stem Cell/Wnt
  3. Large Tumor Suppressor (LATS) E1/E2/E3 Enzyme YAP
  4. DDO-4033

DDO-4033 is a SPOP inhibitor (IC50 = 16.9 μM, Kd = 15.1 μM). DDO-4033 impairs the malignant migration, invasion, and proliferation of clear cell renal cell carcinoma (ccRCC) cell lines. DDO-4033 disrupts SPOP recruitment to its substrate LATS1, inhibits its polyubiquitination and subsequent degradation, and upregulates LATS1 expression. DDO-4033 has promising antitumor activity and is promising for renal cell carcinoma research.

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

DDO-4033

DDO-4033 Chemical Structure

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Based on 1 publication(s) in Google Scholar

DDO-4033 Related Antibodies

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  • Biological Activity

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Description

DDO-4033 is a SPOP inhibitor (IC50 = 16.9 μM, Kd = 15.1 μM). DDO-4033 impairs the malignant migration, invasion, and proliferation of clear cell renal cell carcinoma (ccRCC) cell lines. DDO-4033 disrupts SPOP recruitment to its substrate LATS1, inhibits its polyubiquitination and subsequent degradation, and upregulates LATS1 expression. DDO-4033 has promising antitumor activity and is promising for renal cell carcinoma research[1].

IC50 & Target

Lats1

 

In Vitro

DDO-4033 shows antiproliferative activity against 786-O, A498, A549, HK-2 cells, with IC50s of 1.1, 0.68, 19.96, 18.78 μM, respectively, but has weak antiproliferative activity against HepG2 cells, with an IC50 > 50 μM[1].
DDO-4033 (0-3 μM, 0-32 h) inhibits SPOP-mediated LATS1 ubiquitination and degradation, downregulates the mRNA level of CTGF and CYR61 genes in A498 cells[1].
DDO-4033 (0-5 μM, 24 h) inhibits the colony formation and impairs the cell migratory capacity in A498 cells[1].
DDO-4033 shows affinity for SPOP under MST, BLI, and ITC detection, with Kds of 15.1, 4.96, 4.94 μM, respectively[1].

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

DDO-4033 Related Antibodies

Western Blot Analysis[1]

Cell Line: A498 cells
Concentration: 0 μM, 0.12 μM, 0.25 μM, 0.5 μM, 1 μM
Incubation Time: 0 h, 1 h, 2 h, 4 h,8 h, 12 h, 16 h, 32 h
Result: Inhibited LATS1 ubiquitination, prolongs the half-life of LATS1, and accumulates LATS1.

Cell Migration Assay [1]

Cell Line: A498 cells
Concentration: 0 μM, 0.5 μM, 1 μM
Incubation Time: 24 h
Result: Inhibited the colony formation and impaired the cell migratory capacity.
In Vivo

DDO-4033 (10-50 mg/kg, intratumorally (i.t.) and peritumorally (PTI), once daily, 3 weeks) inhibits tumor growth, upregulates LATS1 and activates the Hippo pathway in A498-Luc xenograft BALB/c nude mice model[1].

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

Animal Model: A498-Luc (1 × 107 cells containing 25% Matrigel, s.c.) xenograft BALB/c nude mice (female, 18-20 g, 4-6 weeks of age) model[1]
Dosage: 10 mg/kg, 50 mg/kg
Administration: i.t. and PTI, once daily, 3 weeks
Result: Inhibited tumor growth, upregulated LATS1 and activated the Hippo pathway, was low in toxicity and did not change organ tissues.
Molecular Weight

406.39

Formula

C20H18N6O4
Appearance

Solid

Color

White to off-white

SMILES

C#CC1=C([N+]([O-])=O)C=C(C(NC2=CC(C)=NN2C(N3)=NC(CCC)=CC3=O)=O)C=C1
Shipping

Room temperature in continental US; may vary elsewhere.
Storage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 6 months
-20°C 1 month
Solvent & Solubility
In Vitro: 

DMSO : 25 mg/mL (61.52 mM; ultrasonic and warming and heat to 60°C; 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 2.4607 mL 12.3035 mL 24.6069 mL
5 mM 0.4921 mL 2.4607 mL 4.9214 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)

Mass
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Concentration
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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)

C1

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Volume (start)

V1

=
Concentration (final)

C2

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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 (6.15 mM); Suspended solution

    This protocol yields a suspended solution of ≥ 2.5 mg/mL (saturation unknown). Suspended solution can be used for oral and intraperitoneal injection.

    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.
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
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 2.4607 mL 12.3035 mL 24.6069 mL 61.5173 mL
5 mM 0.4921 mL 2.4607 mL 4.9214 mL 12.3035 mL
10 mM 0.2461 mL 1.2303 mL 2.4607 mL 6.1517 mL
15 mM 0.1640 mL 0.8202 mL 1.6405 mL 4.1012 mL
20 mM 0.1230 mL 0.6152 mL 1.2303 mL 3.0759 mL
25 mM 0.0984 mL 0.4921 mL 0.9843 mL 2.4607 mL
30 mM 0.0820 mL 0.4101 mL 0.8202 mL 2.0506 mL
40 mM 0.0615 mL 0.3076 mL 0.6152 mL 1.5379 mL
50 mM 0.0492 mL 0.2461 mL 0.4921 mL 1.2303 mL
60 mM 0.0410 mL 0.2051 mL 0.4101 mL 1.0253 mL
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DDO-4033 Related Classifications

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.

Keywords:

DDO-4033DDO4033DDO 4033Large Tumor Suppressor (LATS)E1/E2/E3 EnzymeYAPLarge Tumor SuppressorE1 activating enzymeE2 conjugating enzymeE3 ligating enzymeUbiquitin activating enzymeUbiquitin conjugating enzymeUbiquitin ligaseYes-associated proteinInhibitorRenal cell carcinomaA498 cellsBALB/c nude miceinhibitorinhibit

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