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Chemical Structure| 555-60-2 Chemical Structure| 555-60-2

Structure of CCCP
CAS No.: 555-60-2

Chemical Structure| 555-60-2

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CCCP can inhibit the activation of STING and is widely used as a protonophore uncoupling agent for oxidative phosphorylation. CCCP is the most commonly used inducer of mitophagy in mammalian cells and is thought to exert its effects by reducing the mitochondrial membrane potential.

Synonyms: Carbonyl cyanide 3-chlorophenylhydrazone; Carbonyl Cyanide m-Chlorophenylhydrazone; Mesoxalonitrile 3-chlorophenylhydrazone

4.5 *For Research Use Only! Not for Human Use. We Do Not Sell to Patients.

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Liu, Qinwen ; Chen, Haojie ; Li, Xiang ; Liu, Jingxin ; Li, Yiwen ; Shi, Zhenyi , et al.

Abstract: Triple-negative breast cancer (TNBC) is an aggressive subtype with poor prognosis. Here, we identify , a sesquiterpene lactone from species, as a potent inhibitor of TNBC cell growth with minimal toxicity to normal cells. Transcriptomic analyses revealed that activates , evidenced by elevated , lipid peroxidation, and Fe2+ accumulation, together with GSH depletion and downregulation of SLC7A11 and GPX4. Target identification by drug affinity responsive target stability and mass spectrometry uncovered CDGSH iron sulfur domain 1 (CISD1) as the direct binding partner of . Cellular thermal shift assay, surface plasmon resonance, and dynamics simulations consistently demonstrated that tryptophan-75 is the critical residue mediating this interaction. Functionally, promotes CISD1 ubiquitination and proteasomal degradation, thereby disrupting mitochondrial iron homeostasis and inducing . CISD1 destabilization further impaired mitochondrial integrity and activated PINK1/Parkin-dependent , establishing a dual ferroptosis–mitophagy mechanism. Importantly, genetic knockdown of CISD1 markedly attenuated the anticancer activity of , confirming its essential role. In an orthotopic TNBC mouse model, significantly suppressed tumor growth without causing systemic toxicity. Collectively, our findings provide the first demonstration that directly targets CISD1 at the Trp-75 site to trigger and , highlighting its promise as a therapeutic candidate for TNBC.

Keywords: CISD1 ; ; mitochondrial ; triple-negative breast cancer ; Xanthatin

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Lin, Jin-Yi ; Huang, Ze-Bo ; Zhang, Shi-Qi ; Wu, Yong ; Cheng, Ling-Jun ; Gao, Xiangzheng , et al.

Abstract: PINK1 is a master regulator of PINK1-parkin-mediated , a key process for maintaining mitochondrial homeostasis. The precise regulation of PINK1 is therefore essential for orchestrating . While proteolytic processing of PINK1 and degradation of cleaved PINK1 via the N-end rule under basal conditions have been extensively characterized, the mechanisms governing full-length PINK1 degradation upon mitochondrial damage remain enigmatic. Here, we demonstrate that PINK1 undergoes ubiquitination and proteasomal degradation during through the coordinated action of STUB1 and VCP/p97. Depletion of STUB1 stabilizes full-length PINK1, which paradoxically impairs through the acceleration of parkin degradation. At the organismal level, the STUB1-VCP axis plays an important role in neuronal mitophagy-related memory and learning capacities in the roundworm C. elegans. Congruently, this axis is impaired in the postmortem brain tissues from patients with Alzheimer’s disease compared with cognitively normal controls. Collectively, our findings support STUB1-VCP as a molecular calibrator that fine-tunes full-length PINK1 levels to enable efficient and maintain mitochondrial homeostasis.

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Alternative Products

Product Details of CCCP

CAS No. :555-60-2
Formula : C9H5ClN4
M.W : 204.62
SMILES Code : N#CC(C#N)=NNC1=CC=CC(Cl)=C1
Synonyms :
Carbonyl cyanide 3-chlorophenylhydrazone; Carbonyl Cyanide m-Chlorophenylhydrazone; Mesoxalonitrile 3-chlorophenylhydrazone
English Name :(3-Chlorophenyl)carbonohydrazonoyl dicyanide
MDL No. :MFCD00001848
InChI Key :UGTJLJZQQFGTJD-UHFFFAOYSA-N
Pubchem ID :2603

Safety of CCCP

Related Pathways of CCCP

PI3K-AKT

Isoform Comparison

Biological Activity

In Vitro:

Cell Line
Concentration Treated Time Description References
GV oocytes 1 µM 0.5 hours CCCP treatment activated the PRKN-mediated mitophagy pathway and resulted in meiotic arrest at the MI stage Autophagy. 2022 Mar;18(3):643-660.
Wheat root cells 10 µM 1 hour To study the effect of CCCP on cytokinin accumulation in root cells, the results showed that CCCP treatment decreased cytokinin accumulation in root cells but increased both flow from the roots and accumulation in the shoots. J Exp Bot. 2014 Jun;65(9):2287-94.
MRC5 cells 25 µM 2 hours CCCP treatment significantly increased mitolysosome formation and promoted VZV replication but attenuated IFN production. Cell Death Dis. 2024 Jan 6;15(1):16.
HaCaT cells 25 µM 2 hours CCCP treatment significantly increased mitolysosome formation and promoted VZV replication but attenuated IFN production. Cell Death Dis. 2024 Jan 6;15(1):16.
293T cells 10 µM 2 hours CCCP treatment induced mitochondrial damage, manifested as mitochondrial membrane rupture and vacuolization Autophagy. 2022 Mar;18(3):643-660.
SH-SY5Y cells 20 µM 2 hours To investigate the role of AMBRA1 in the PINK1-PRKN signaling pathway after CCCP treatment, results showed that AMBRA1 downregulation leads to reduced PINK1 stability and mitophagy. Autophagy. 2022 Aug;18(8):1752-1762.
HeLa cells 20 µM 2 hours To investigate the role of AMBRA1 in the PINK1-PRKN signaling pathway after CCCP treatment, results showed that AMBRA1 downregulation leads to reduced PINK1 stability and mitophagy. Autophagy. 2022 Aug;18(8):1752-1762.
Yeast cells 2 µM 20 minutes To test the direct inhibitory effect of CCCP on TbAQP2 glycerol permeability, results showed that CCCP directly inhibits TbAQP2 glycerol permeability in the single-digit micromolar range. Cells. 2020 Oct 21;9(10):2335.
Human induced pluripotent stem cell-derived dopaminergic neurons 10 µM 48 hours To evaluate the effect of CCCP-induced mitochondrial damage on healthy neurons, it was found that healthy neurons responded to CCCP by clearing impaired mitochondria, and this process was accelerated by USP30 inhibition. Cell Death Dis. 2024 Jan 15;15(1):52.
PARK2 knockout neurons 10 µM 48 hours To evaluate the effect of CCCP-induced mitochondrial damage on PARK2 knockout neurons, it was found that PARK2 knockout neurons showed an impaired mitophagic response to CCCP, but USP30 inhibition promoted mitophagy in PARK2 knockout neurons. Cell Death Dis. 2024 Jan 15;15(1):52.
Vascular smooth muscle cells (A10) 2 µM 5 minutes and 20 minutes CCCP increased the cellular ADP/ATP ratio and activated AMPK after 5 and 20 minutes. Br J Pharmacol. 2016 Nov;173(21):3145-3158.
GT1-7 cells 10 µM 6 hours CCCP was used for mitochondrial depolarization to assess its impact on the mitochondrial network. Results showed that CCCP led to a significant decrease in mitochondrial volume, indicating that the network likely underwent fragmentation prior to observation. Cells. 2023 Jun 27;12(13):1726.

In Vivo:

Species
Animal Model
Administration Dosage Frequency Description References
Mice Aged mice Intraperitoneal injection 1 µM Every other day for one month ML098 treatment improved oocyte quality and fertility in aged mice Autophagy. 2022 Mar;18(3):643-660.
Rat POI model Intraperitoneal injection 2 mg/kg Every other day for 15 days CCCP inhibited the ability of BMSCs-MOX treatment to regulate mitophagy and exacerbated Cy-induced ovarian injury. Stem Cell Res Ther. 2024 Apr 8;15(1):102

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