Cat. No.: HY-145669

DIF-3

8 Cited Publications

Customer Review

Based on 8 publication(s) in Google Scholar

DIF-3 is an orally active anticancer agent. DIF-3 reduces the expression levels of cyclin D1 and c-Myc by facilitating their degradation via activation of GSK-3β. DIF-3 inhibits Wnt/β-catenin signaling pathway-related proteins in cells. DIF-3 induces reactive oxygen species (ROS) and autophagy. DIF suppresses the growth of Trypanosoma. cruzi in HT1080 cells. DIF-3 exerts antitumor effects both in vitro and in vivo.

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

Purity: 99.84%
CAS No.: 113411-17-9
Formula: C₁₃H₁₇ClO₄
Molecular Weight:272.72

Storage:

4°C, protect from light

* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

DIF-3 Chemical Structure

CAS No.: 113411-17-9

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Top Publications Citing

Use of MCE Products

Fluorescent Dye

Strong Fluorescence Signal / Multi-Disciplinary Applications / High Fluorescence Specificity

Biological Activity
- Description
- Cellular Effect
- In Vitro
- In Vivo
Chemical Information
Publications (8)
Solvent & Solubility
- In Vitro
- In Vivo
Purity & Documentation
References
Help & FAQs
Bioz

Biological Activity

Description

All Wnt Isoforms :

All CDK Isoforms :

All GSK-3 Isoforms :

Cellular Effect

Cell Line	Type	Value	Description	References
A549	IC₅₀	2 μM Compound: DIF-3	Growth inhibition of human A549 cells Growth inhibition of human A549 cells	[PMID: 28814374]
S2	IC₅₀	4.6 μM Compound: 2, DIF-3	Immunosuppressive activity in Drosophila S2 cells expressing att-luc reporter gene assessed as inhibition of peptidoglycan-induced attacin production pre-treated for 1.5 hrs before peptidoglycan stimulation for 8 hrs by luciferase reporter gene assay Immunosuppressive activity in Drosophila S2 cells expressing att-luc reporter gene assessed as inhibition of peptidoglycan-induced attacin production pre-treated for 1.5 hrs before peptidoglycan stimulation for 8 hrs by luciferase reporter gene assay	[PMID: 26122773]

In Vitro

DIF-3 (10-30 μM, 24-48 h) inhibits proliferation in a dose-dependent manner in human colon cancer cells^[1].
DIF-3 (30 μM, 24 h) arrests the cell cycle at G₀/G₁ phase in human colon cancer cells^[1].
DIF-3 (10-30 μM, 1-24 h) induces proteolysis of cyclin D1 and c-Myc in human colon cancer cells via activation of GSK-3β^[1].
DIF-3 (10-30 μM, 1-24 h) inhibits Wnt/β-catenin signaling pathway-related proteins in human colon cancer cells^[1].
DIF-3 (10 μM, 4 h) inhibits lysophosphatidic acide (LPA) stimulated migration of murine osteosarcoma LM8 cells^[2].
DIF-3 (20 μM, 0.25-24 h) inhibits mTOR signaling and induces autophagy in K562 CML cells^[3].
DIF-3 (20 μM) induces mitochondrial membrane potential (MMP) (0-10 min), a decrease of ATP content (6 h) and an induction of ROS production (0-30 min) in K562 cells^[3].
DIF-3 (10 μM, 3 d) suppresses the growth of Trypanosoma. cruzi in HT1080 cells with an IC₅₀ of 3.95 μM^[4].

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

Cell Proliferation Assay^[1]

Cell Line:

HCT-116, DLD-1
Concentration:

10, 20, 30 μM
Incubation Time:

24, 48 h
Result:

Strongly inhibited proliferation in a dose-dependent manner in both cell lines.
HCT-116 cells were more sensitive to DIF-3 than DLD-1 cells.

Cell Cycle Analysis^[1]

Cell Line:

HCT-116, DLD-1
Concentration:

30 μM
Incubation Time:

24 h
Result:

Significantly increased the number of cells in G0/G1 phase and decreased those in S phase in both cell lines.
Also significantly increased the number of cells in G2 phase when HCT-116 cells were employed, this effect was not observed by using DLD-1 cells.

Cell Migration Assay ^[2]

Cell Line:

LM8
Concentration:

10 μM
Incubation Time:

4 h
Result:

Suppressed LPA-induced cell migration, whereas did not significantly affected cell migration in the absence of LPA.

Cell Autophagy Assay^[3]

Cell Line:

K562
Concentration:

20 μM
Incubation Time:

0.25, 0.5, 1, 3, 6, 24 h for mTOR study; 6, 16, 36, 48 h for Cathepsin B activity study
Result:

Induced the rapid dephosphorylation of mTOR at Ser2481. Inhibited the mTOR pathway and leads to a rapid and robust conversion of LC3-I to LC3-II.
Increased autophagy was accompanied by the late activation of cathepsin B.

Western Blot Analysis^[1]

Cell Line:

HCT-116
Concentration:

10, 20, 30 μM
Incubation Time:

1, 3, 6, 12, 24 h
Result:

Induced a rapid and marked reduction in the amount of cyclin D1 protein and c-Myc protein in a time and dose-dependent manner.
Had no effect on the expression level of β-catenin (30 μM for 24 h).
Reduced the amount of TCF7L2 which increased gradually over time in control cells, in a time and dose-dependent manner.
Reduction of TCF7L2 was associated with the transcriptional inhibition of cyclin D1 mRNA in HCT-116 cells.

Western Blot Analysis^[1]

Cell Line:

HCT-116
Concentration:

30 μM
Incubation Time:

1 h
Result:

Effects could be attenuated by MG132 (HY-13259) and SB216763 (HY-12012), indicating the involvement of GSK-3β in degradation of cyclin D1 and c-Myc.

In Vivo

DIF-3 (150 mg/kg; p.o.; once a day for 5 days/week over 4 weeks) inhibits oxidative stress (0.2% KBrO₃)-induced tumors in Mutyh^-/- mice^[1].

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

Animal Model:

Mutyh^-/- mice (KBrO3 dissolved in water at a concentration of 2 g/L was administered to 4-week-old mice for 12 weeks)^[1]
Dosage:

150 mg/kg
Administration:

Oral gavage (p.o.); once a day for 5 days/week over 4 weeks
Result:

Markedly reduced the number of intestinal tumors, especially the number of large tumors with a diameter of >2.0 mm.
No differences in the appearance, activity, body weight, or blood cell counts.
The numbers of TCF7L2-and cyclin D1-positive nuclei in tumors were significantly decreased.

Chemical Information

CAS No. 113411-17-9
Appearance Solid
Molecular Weight 272.72
Formula C₁₃H₁₇ClO₄
Color White to off-white
SMILES

O=C(CCCCC)C1=C(C=C(C(Cl)=C1O)OC)O
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

4°C, protect from light

* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

Publications (8)

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Journal Impact Factor

J Transl Med

MaR1 and NGF combine to inhibit autophagy through the GSK-3β/β-catenin pathway to promote sciatic nerve repair. [Abstract]

2026 Feb 9;24(1):361. PMID: 41664203
Oncogene

D-mannose induces TFE3-dependent lysosomal degradation of EGFR and inhibits the progression of NSCLC. [Abstract]

2023 Nov;42(47):3503-3513. PMID: 37845392
Mol Med

CGREF1 modulates osteosarcoma proliferation by regulating the cell cycle through the Wnt/β-catenin signaling pathway. [Abstract]

2024 Dec 20;30(1):260. PMID: 39707194
Int J Oncol

Ephrin‑B2 promotes gastric cancer growth by inhibiting apoptosis and regulating the cell cycle via the Wnt/β‑catenin signaling pathway. [Abstract]

2026 Jan;68(1):8. PMID: 41268595
mBio

The Streptococcus pyogenes Rgg2/Rgg3 quorum sensing system causes global suppression of macrophage inflammatory programs via an intranuclear mechanism. [Abstract]

2025 Sep 25:e0037325. PMID: 40996031
Diabetol Metab Syndr

IL-37 inhibited inflammation to improve gestational diabetes mellitus through the GSK3/NF-κB pathway. [Abstract]

2025 Jun 5;17(1):193. PMID: 40474280
Placenta

The downregulation of B2R inhibits spiral artery remodeling by reducing the autophagy of trophoblast cells. [Abstract]

2025 Oct 7:171:194-204. PMID: 41076744
bioRxiv

Suppressed macrophage response to quorum-sensing-active Streptococcus pyogenes occurs at the level of the nucleus. [Abstract]

2025 Feb 8:2025.02.07.637189. PMID: 39975246

Solvent & Solubility

In Vitro:

DMSO : 100 mg/mL (366.68 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

View the Complete Stock Solution Preparation Table

Preparing Stock Solutions

View the Complete Stock Solution Preparation Table

Concentration Solvent Mass

1 mg

5 mg

10 mg

Concentration

Solvent

Mass

1 mM

3.6668 mL

18.3338 mL

36.6676 mL

5 mM

0.7334 mL

3.6668 mL

7.3335 mL

10 mM

0.3667 mL

1.8334 mL

3.6668 mL

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 (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Molarity Calculator

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

Mass Mass

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Molecular Weight Molecular Weight

Dilution Calculator

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

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In Vivo:

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

Stock Solution Working Solution

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% Corn Oil

Solubility: ≥ 2.5 mg/mL (9.17 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:

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mg/kg
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g
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μL
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Please enter your animal formula composition:

%
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Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
%
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+
%
Tween-80 + +
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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).

Working solution concentration: 0.22 mg/mL

Purity & Documentation

Select Batch:

Purity: 99.98%

Assay: 99.98%

ee.: 99.98%

Data Sheet (282 KB)
SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Portuguese - PT (252 KB)
COA (2659 KB)
Elemental Analysis Report (2659 KB)
HNMR (2659 KB)
CNMR (2659 KB)
RP-HPLC (2659 KB)
NP-HPLC (2659 KB)
MS (2659 KB)
LCMS (2659 KB)
IR (2659 KB)
GC (2659 KB)
OR (2659 KB)
SDS-PAGE (2659 KB)
SEC-HPLC (2659 KB)
Other Report (2659 KB)
Handling Instructions (2659 KB)

References

[1]. Naoya Kubokura , et al. Differentiation-inducing factor-3 inhibits intestinal tumor growth in vitro and in vivo. J Pharmacol Sci. 2015 Apr;127(4):446-55. [Content Brief]

[2]. Kubohara Y, et al. Derivatives of Dictyostelium differentiation-inducing factors inhibit lysophosphatidic acid-stimulated migration of murine osteosarcoma LM8 cells. Biochem Biophys Res Commun. 2015 Aug 7;463(4):800-5. [Content Brief]

[3]. Dubois A, et al. Differentiation inducing factor 3 mediates its anti-leukemic effect through ROS-dependent DRP1-mediated mitochondrial fission and induction of caspase-independent cell death. Oncotarget. 2016 May 3;7(18):26120-36. [Content Brief]

[4]. Nakajima-Shimada J, et al. Derivatives of Dictyostelium discoideum differentiation-inducing factor-3 suppress the activities of Trypanosoma cruzi in vitro and in vivo. Biochem Pharmacol. 2013 Jun 1;85(11):1603-10. [Content Brief]

Help & FAQs

How should lyophilized recombinant proteins be reconstituted and stored?

1. Before opening the cap, centrifuge the vial at 13000 rpm for 20-30 seconds. This step will ensure that any lyophilized powder that may have adhered to the cap or walls is collected at the bottom of the vial, minimizing the risk of product loss.

Complete Stock Solution Preparation Table

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg
DMSO	1 mM	3.6668 mL	18.3338 mL	36.6676 mL	91.6691 mL
	5 mM	0.7334 mL	3.6668 mL	7.3335 mL	18.3338 mL
	10 mM	0.3667 mL	1.8334 mL	3.6668 mL	9.1669 mL
	15 mM	0.2445 mL	1.2223 mL	2.4445 mL	6.1113 mL
	20 mM	0.1833 mL	0.9167 mL	1.8334 mL	4.5835 mL
	25 mM	0.1467 mL	0.7334 mL	1.4667 mL	3.6668 mL
	30 mM	0.1222 mL	0.6111 mL	1.2223 mL	3.0556 mL
	40 mM	0.0917 mL	0.4583 mL	0.9167 mL	2.2917 mL
	50 mM	0.0733 mL	0.3667 mL	0.7334 mL	1.8334 mL
	60 mM	0.0611 mL	0.3056 mL	0.6111 mL	1.5278 mL
	80 mM	0.0458 mL	0.2292 mL	0.4583 mL	1.1459 mL
	100 mM	0.0367 mL	0.1833 mL	0.3667 mL	0.9167 mL

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DIF-3

Anti-Infection Compound Library

Cell Cycle/DNA Damage Compound Library

Kinase Inhibitor Library

PI3K/Akt/mTOR Compound Library

Protein Tyrosine Kinase Compound Library

Stem Cell Signaling Compound Library

Wnt/Hedgehog/Notch Compound Library

Anti-Cancer Compound Library

Anti-Aging Compound Library

Differentiation Inducing Compound Library

Oxygen Sensing Compound Library

Glycolysis Compound Library

Cytoskeleton Compound Library

Orally Active Compound Library

Neuroprotective Compound Library

Anti-Breast Cancer Compound Library

Anti-Pancreatic Cancer Compound Library

Anti-Blood Cancer Compound Library

Anti-Cancer Metabolism Compound Library

Glucose Metabolism Compound Library

Anti-Liver Cancer Compound Library

Anti-Colorectal Cancer Compound Library

Anti-Prostate Cancer Compound Library

Anti-Pulmonary Fibrosis Compound Library

Osteogenesis Compound Library

Cancer Stem Cells Compound Library

Antihypertensive Compound Library

Metabolic Enzyme Compound Library

Serine/Threonine Kinase Inhibitor Library

Extracellular Vesicles (EVs) Compound Library

Anti-Hematopathy Compound Library

Anti-Ovarian Cancer Compound Library

Multi-Target Compound Library

Bioactive Compound Library Max

MCE Bioactive Compound Library

Bioactive Compound Library

Anti-Gastric Cancer Compound Library

Anti-Fibrosis Compound Library

Protein Kinase Compound Library

Cell Proliferation Compound Library

High-Throughput Bioactive Compound Library

Anti-Rheumatic Arthritis Compound Library

Posttranslational Modification Library

High-Efficiency Gene Editing Compound Library

Publications Citing Use of MedChemExpress (MCE) DIF-3

All Wnt Isoforms

All CDK Isoforms

All GSK-3 Isoforms

Biological Activity

Chemical Information

Publications (8)

Solvent & Solubility

Purity & Documentation

References

Help & FAQs

Complete Stock Solution Preparation Table

DIF-3 Related Classifications

Cat. No.: HY-145669