Cat. No.: HY-189265

Antitumor agent-226

Antitumor agent-226 is a DNMT3A inhibitor and epigenetic modulator. Antitumor agent-226 inhibits cell proliferation and migration by downregulating DNA Methyltransferase, reducing global DNA methylation, and modulating JNK, ERK1/2, and p38 phosphorylation through the MAPK pathway to induce mitochondria-dependent apoptosis, G2/M phase arrest, autophagy, and ROS production. Antitumor agent-226 can be used for research on prostate cancer.

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

CAS No.: 3056235-77-6
Formula: C₂₅H₂₁ClF₂N₄O₅
Molecular Weight:530.91

Storage:

Please store the product under the recommended conditions in the Certificate of Analysis.

Antitumor agent-226 Chemical Structure

CAS No.: 3056235-77-6

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Use of MCE Products

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

Biological Activity
- Description
- IC₅₀ & Target
- Cellular Effect
- In Vitro
- In Vivo
Chemical Information
Purity & Documentation
References
Calculators
Help & FAQs
Classifications
Bioz

Biological Activity

Description

IC₅₀ & Target

^[1]

DNMT3A

ERK1

ERK2

JNK

p38 MAPK

All DNA Methyltransferase Isoforms :

All p38 MAPK Isoforms :

All JNK Isoforms :

All ERK Isoforms :

Cellular Effect

Cell Line	Type	Value	Description	References
A549	IC50	1.58 μM	Antiproliferative activity against human A549 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay. Antiproliferative activity against human A549 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay.	42612263
PC-3	IC50	1.35 μM	Antiproliferative activity against human PC-3 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay. Antiproliferative activity against human PC-3 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay.	42612263
T47D	IC50	1.55 μM	Antiproliferative activity against human T47D cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay. Antiproliferative activity against human T47D cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay.	42612263
SMMC-7721	IC50	1.79 μM	Antiproliferative activity against human SMMC-7721 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay. Antiproliferative activity against human SMMC-7721 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay.	42612263
HEK293	IC50	11.22 μM	Cytotoxicity against human HEK293 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay. Cytotoxicity against human HEK293 cells assessed as reduction in cell viability incubated for 48 hrs by MTT assay.	42612263

In Vitro

Anti-cancerCell Cycle RegulationApoptosisAutophagyMitochondrial FunctionOxidative StressOthers

Anti-cancer

Antitumor agent-226 (compound 11M) (0.625-10 μM; 48 h) exhibits the strongest antiproliferative activity against A549 and PC-3 cell lines, with IC₅₀ values of 1.58 μM and 1.51 μM, respectively, and shows the highest selectivity index in PC-3 cells (SI = 8.31)^[1].
Antitumor agent-226 (1-3 μM; 24 h) inhibits PC-3 cell proliferation in a dose-dependent manner and induces morphological changes characteristic of cell death^[1].
Antitumor agent-226 (1-3 μM; 24 h) significantly inhibits the colony-forming ability of PC-3 cells^[1].
Antitumor agent-226 (1-3 μM; 24-48 h) significantly inhibits the migration of PC-3 cells^[1].
Antitumor agent-226 (1-3 μM) combined with Acetylcysteine (NAC) (HY-B0215) restores the viability of PC-3 cells^[1].

Anti-cancer

Cell Cycle Regulation

Antitumor agent-226 (1-3 μM; 24 h) induces G2/M phase cell cycle arrest in PC-3 cells^[1].
Antitumor agent-226 (1-3 μM; 24 h) induces G2/M phase arrest in PC-3 cells by upregulating p21 and downregulating Cyclin B1 and CDK1^[1].

Anti-cancer

Apoptosis

Antitumor agent-226 (1-3 μM; 24 h) induces apoptosis in PC-3 cells through downregulation of Bcl-2 and Bax^[1].

Apoptosis

Antitumor agent-226 (1-3 μM; 24 h) activates the MAPK signaling pathway in PC-3 cells by increasing the phosphorylation of JNK, ERK1/2, and p38^[1].
Antitumor agent-226 (1-3 μM; 24 h) induces apoptotic nuclear morphological changes in PC-3 cells^[1].
Antitumor agent-226 (1-3 μM; 24 h) induces apoptosis in PC-3 cells in a dose-dependent manner^[1].
Antitumor agent-226 (1-3 μM) induces apoptosis in PC-3 cells, and inhibition of autophagy promotes this apoptotic effect^[1].

Autophagy

Antitumor agent-226 (1-3 μM; 24 h) induces autophagy in PC-3 cells by increasing LC3-II expression and promoting the conversion of LC3-I to LC3-II^[1].
Antitumor agent-226 (3 μM; 24 h) induces autophagy in PC-3 cells, manifested by the formation of autophagosomes^[1].
Antitumor agent-226 (1-3 μM) increases the expression level of LC3-II in PC-3 cells; combined treatment with the autophagy inhibitor 3-Methyladenine (3-MA) (HY-19312) reverses this effect^[1].

Mitochondrial Function

Antitumor agent-226 (1-3 μM; 24 h) induces a dose-dependent decrease in mitochondrial membrane potential in PC-3 cells^[1].

Oxidative Stress

Antitumor agent-226 (1-3 μM; 24 h) induces ROS production in PC-3 cells^[1].

Cell Cycle Regulation

Apoptosis

Antitumor agent-226 (1-3 μM) modulates the expression of cell cycle regulatory proteins P21, CyclinB1, and CDK1 in prostate cancer cells^[1].
Antitumor agent-226 (1-3 μM) alters the expression of apoptosis-related proteins Bax and Bcl-2 in prostate cancer cells^[1].
Antitumor agent-226 (1-3 μM) affects the MAPK signaling pathway by modulating the phosphorylation of JNK, ERK, and P38 in prostate cancer cells^[1].
Antitumor agent-226 (1-3 μM) induces autophagy in prostate cancer cells, manifested as modulation of LC3-I and LC3-II levels^[1].
Antitumor agent-226 (1-3 μM) downregulates DNMT3A expression in prostate cancer cells^[1].

Others

Antitumor agent-226 (1-3 μM; 24 h) significantly downregulates DNMT3A protein levels in PC-3 cells^[1].
Antitumor agent-226 (3 μM; 24 h) significantly alters gene expression in PC-3 cells, particularly affecting metabolic processes, intracellular structures, and the MAPK signaling pathway^[1].
Antitumor agent-226 (3 μM; 24 h) reduces global DNA methylation levels in PC-3 cells, leading to genome-wide hypomethylation^[1].

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

Cell Viability Assay^[1]

Cell Line:

PC-3
Concentration:

1, 1.5, 2, 2.5, 3 μM
Incubation Time:

24 h
Result:

Caused significant morphological changes including cell shrinkage, fragmentation, rounding, and detachment.
Decreased cell viability gradually with increasing concentrations.

Cell Proliferation Assay^[1]

Cell Line:

PC-3
Concentration:

1, 2, 3 μM
Incubation Time:

24 h
Result:

Induced a marked reduction in colony numbers compared to the untreated group.

Cell Migration Assay ^[1]

Cell Line:

PC-3
Concentration:

1, 2, 3 μM
Incubation Time:

24, 48 h
Result:

Significantly inhibited PC-3 cell migration at 0, 24, and 48 h.
Strongly inhibited migration at concentrations of 2 μM and 3 μM.

Cell Cycle Analysis^[1]

Cell Line:

PC-3
Concentration:

1, 2, 3 μM
Incubation Time:

24 h
Result:

Increased the proportion of cells in G2/M phase to 19.7% at 1 μM.
Increased the proportion of cells in G2/M phase to 24.8% at 2 μM.
Increased the proportion of cells in G2/M phase to 27.4% at 3 μM.

Western Blot Analysis^[1]

Cell Line:

PC-3
Concentration:

1, 3 μM
Incubation Time:

24 h
Result:

Markedly upregulated the expression of p21.
Downregulated the expression of Cyclin B1 and CDK1.\nDownregulated the expression of apoptosis-related proteins Bcl-2 and Bax.\nSignificantly increased the phosphorylation levels of JNK, ERK1/2, and p38, while total protein levels remained unchanged.\nIncreased LC3-II expression.
Promoted the conversion of LC3-I to LC3-II.\nSignificantly reduced DNMT3A protein levels.

Immunofluorescence^[1]

Cell Line:

PC-3
Concentration:

1, 2, 3 μM
Incubation Time:

24 h
Result:

Induced intense, concentrated blue fluorescence in nuclei, a hallmark of early apoptosis, which became more pronounced with increasing concentrations.

Cell Cytotoxicity Assay^[1]

Cell Line:

PC-3
Concentration:

1, 2, 3 μM
Incubation Time:

24 h
Result:

Reduced mitochondrial membrane potential in 28.83% of cells at 1 μM.
Reduced mitochondrial membrane potential in 42.21% of cells at 2 μM.
Reduced mitochondrial membrane potential in 81.11% of cells at 3 μM.\nSignificantly increased ROS levels compared to the control group.

Apoptosis Analysis^[1]

Cell Line:

PC-3
Concentration:

1, 2, 3 μM
Incubation Time:

24 h
Result:

Induced apoptosis rates of 8.49% at 1 μM.
Induced apoptosis rates of 35.43% at 2 μM.
Induced apoptosis rates of 38.99% at 3 μM.

Cell Autophagy Assay^[1]

Cell Line:

PC-3
Concentration:

3 μM
Incubation Time:

24 h
Result:

Revealed the presence of autophagic vacuoles and autophagosomes, suggesting active autophagy.

In Vivo

Anti-cancer

Antitumor agent-226 (compound 11M) (5 mg/kg; i.p.; every other day; 21 days) significantly inhibits tumor growth in the PC-3 xenograft mouse model, with a favorable safety profile, accompanied by MAPK pathway activation and DNMT3A downregulation^[1].

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

Animal Model:

BALB/c nude mice (male, 5 weeks old, subcutaneous PC-3 cell xenograft model)^[1]
Dosage:

5 mg/kg
Administration:

i.p.; every two days; 21 days
Result:

Significantly reduced tumor volume and weight compared to control and cisplatin groups.
Induced extensive apoptosis and necrosis with nuclear fragmentation and cytoplasmic vacuolation.
Significantly increased apoptosis by TUNEL staining.
Significantly upregulated phosphorylated ERK (p-ERK) levels while total ERK expression remained stable.
Downregulated DNMT3A expression.
Showed no significant weight loss or signs of toxicity.
Showed no significant pathological changes in H&E-stained sections of the heart, liver, spleen, lungs, and kidneys.

Related Protocols :

Chemical Information

CAS No. 3056235-77-6
Molecular Weight 530.91
Formula C₂₅H₂₁ClF₂N₄O₅
SMILES

O=C1C(Cl)=CCCN1C(/C=C/C2=CC=C(OCC3=CN(C4=CC=C(OC(F)F)C=C4)N=N3)C(OC)=C2)=O
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

Please store the product under the recommended conditions in the Certificate of Analysis.

Purity & Documentation

Handling Instructions (2659 KB)

References

[1]. Zhang Y, et al. Novel Piperlongumine derivatives exert multimodal antitumor effects in prostate cancer via MAPK pathway activation, poly-mechanistic apoptosis induction, and DNMT3A-mediated DNA methylation reduction. Bioorganic chemistry. 2026 Oct 05;181:110408.

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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.

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Antitumor agent-226

All DNA Methyltransferase Isoforms

All p38 MAPK Isoforms

All JNK Isoforms

All ERK Isoforms