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Cat. No.: HY-187371

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DA29

DA29 is a human caseinolytic protease P (hClpP) activator, with an EC₅₀ of 0.85 μM and a K_d of 185 nM. DA29 binds to the hydrophobic allosteric pocket of this enzyme, activates its proteolytic function, and displaces hClpX from the hClpXP complex. DA29 induces mitochondrial dysfunction and ROS accumulation. DA29 exerts cytotoxic effects on cancer cells and organoids. DA29 can be used in the research of diffuse intrinsic pontine glioma.

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

Formula: C₂₀H₁₈F₆N₂O
Molecular Weight:416.36

Storage:

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

DA29 Chemical Structure

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Use of MCE Products

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

Biological Activity
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- In Vitro
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Biological Activity

Description

In Vitro

OthersAnti-cancerOxidative StressMitochondrial FunctionLipid Metabolism

Others

DA29 (compound 26) potently activates purified human hClpP, with an EC₅₀ of 0.85 μM and an activation rate of 95% at a concentration of 100 μM in cell-free fluorescence assays; it binds to purified human hClpP with a K_d of 185 nM; and it enhances the thermal stability of hClpP in Caco-2 cells^[1].
DA29 exhibits high basolateral permeability in Caco-2 and MDCK-MDR1 cell monolayer models, with BA/AB ratios of 5.91 and 4.61, respectively, suggesting its potential blood-brain barrier penetration ability; meanwhile, it shows weak interaction with P-gp (EC₅₀ = 71 μM)^[1].

Anti-cancer

DA29 (1-100 μM; 72 h) exhibits cytotoxic activity against the SU-DIPG-36 and SU-DIPG-50 diffuse intrinsic pontine glioma (DIPG) cell lines, with IC₅₀ values of 4.0 μM and 11.6 μM, respectively; it reduces the viability of DIPG tumor organoids DMG2_O and DMG3_O, with IC₅₀ values of 2.57 μM and 1.94 μM, respectively^[1].

Oxidative Stress

DA29 (1 μM; 1-24 h) significantly raises intracellular ROS levels in SU-DIPG-36 cells, with the notable increase detected at 6 h^[1].

Mitochondrial Function

DA29 (1.5 μM; 24 h) displaces ClpX from the ClpXP complex, reduces the levels of specific mitochondrial respiratory chain subunits and TFAM, and decreases mtDNA levels in SU-DIPG-36 cells; it also reduces the levels of mitochondrial ribosomal subunits, inhibits polyribosome formation, and disassembles the ClpXP complex^[1].

Lipid Metabolism

DA29 (24 h) remodels the lipid profiles of SU-DIPG-36 and SH-SY5Y cells, reduces most lysophosphatidylcholine (LPC) and sphingomyelin (SM) species, and alters the species distribution of triglyceride (TG), which is consistent with the characteristics of impaired mitochondrial function^[1].

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

Western Blot Analysis^[1]

Cell Line:

SU-DIPG-36 DIPG cell line
Concentration:

1.5 μM
Incubation Time:

24 h
Result:

Completely abolished ClpX levels while leaving ClpP unaffected, indicating ClpXP complex disassembly.
Reduced levels of SDHB (Complex II), UQCRC2 (Complex III), ATP5A (Complex V), and COI (Complex IV), but not NDUFB8 (Complex I).
Strongly reduced TFAM levels and dramatically decreased mtDNA levels.
Left VDAC1 (mitochondrial mass marker) unchanged.

Related Protocols :

Chemical Information

Molecular Weight 416.36
Formula C₂₀H₁₈F₆N₂O
SMILES

O=C1N(CC2=CC=C(C(F)(F)F)C=C2)CCN(CC3=CC=CC(C(F)(F)F)=C3)C1
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]. Miciaccia M, et al. Dysregulation of Human ClpP Using Small Molecules with Piperazine-Based Scaffold for Diffuse Intrinsic Pontine Glioma Therapy Validated by Patient-Derived Tumor Organoids. J Med Chem. 2026 Jul 8.

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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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