Dichloroacetophenone(2,2-) / ROS Cancer Research Results

DAP, Dichloroacetophenone(2,2-): Click to Expand ⟱
Features:
2,2-Dichloroacetophenone (DAP) is an acetophenone derivative characterized by the presence of two chlorine atoms on the aromatic ring, typically at the 2‑position relative to the carbonyl group. Its structure and properties differ significantly from compounds like dichloroacetate (DCA).
-DAP may trigger the production of reactive oxygen species (ROS) in cells.
-Increased ROS can lead to cellular damage, which may trigger apoptosis (programmed cell death) in cancer cells that are already under metabolic and oxidative stress.
-Cytotoxic effects via DNA damage.

*** Importantly, 2,2-dichloroacetophenone (DAP) is a much more potent inhibitor of PDK1(Than DCA). It is effective at concentrations in the micromolar (μM) range.
- poor aqueous solubility and lower stability of DAP limits its therapeutic application

2,2-Dichloroacetophenone — also called α,α-dichloroacetophenone, is a chlorinated aromatic ketone and experimental small-molecule inhibitor of pyruvate dehydrogenase kinase 1. It is classified as a preclinical metabolic anticancer agent and chemical research compound. DAP is structurally and pharmacologically distinct from dichloroacetate and should also be distinguished from α-chloroacetophenone, the riot-control agent commonly abbreviated CN. DAP has shown activity against acute myeloid leukemia, EGFR-mutant non-small-cell lung cancer, and prostate-cancer models, but poor aqueous solubility, limited stability, weak selectivity, and insufficient pharmacokinetic characterization restrict its translational potential.

Primary mechanisms (ranked):

  1. Inhibition of pyruvate dehydrogenase kinase 1, relieving inhibitory control of the pyruvate dehydrogenase complex and disrupting tumor glycolytic and mitochondrial energy metabolism.
  2. Destabilization of PDK1-associated survival proteins, including PI3K, Akt, ULK1, and BCL-xL, partly through altered interaction with the CBL-b ubiquitin ligase system.
  3. Induction of mitochondrial apoptosis through BAX activation, caspase-3 cleavage, PARP cleavage, and suppression of BCL-2 and BCL-xL.
  4. Suppression of PI3K/Akt/mTOR prosurvival signaling.
  5. Suppression of cytoprotective autophagy through reduced ULK1, Beclin-1, LC3-II, ATG5, and ATG7 signaling.
  6. Inhibition of proliferation, clonogenic survival, migration, and tumor growth, with reported enhancement of EGFR-tyrosine-kinase inhibitor activity and activity against some drug-resistant cancer models.

Bioavailability / PK relevance: Human pharmacokinetic data are unavailable. Free DAP has poor aqueous solubility and limited physicochemical stability, and later medicinal-chemistry studies describe weak anticancer potency and poor selectivity relative to optimized derivatives. Lactoferrin nanoparticle encapsulation improved dispersion, stability, tumor-cell targeting, and experimental antitumor efficacy, but this remains a preclinical delivery strategy.

In-vitro vs systemic exposure relevance: Most free-DAP experiments used approximately 5–100 µM, commonly around 20–40 µM. No validated human plasma concentration or clinically achievable exposure has been established. Nanoparticle-formulated DAP showed activity at lower in-vitro concentrations, including approximately 1 µM, and at 20 mg/kg in a mouse xenograft model, but these results cannot be extrapolated to human systemic exposure.

Clinical evidence status: Preclinical only. Evidence consists of cancer-cell studies and mouse xenograft experiments in acute myeloid leukemia, non-small-cell lung cancer, and prostate cancer. No completed human anticancer trial, approved therapeutic formulation, established clinical dose, or regulatory approval was identified.


Mechanistic Effects of 2,2-Dichloroacetophenone

Rank Pathway / Axis Cancer Cells Normal Cells TSF Primary Effect Notes / Interpretation
1 Pyruvate dehydrogenase kinase 1 PDK1 ↓ PDK1 may ↓ R–G Disrupts metabolic adaptation and tumor-cell survival DAP is reported to inhibit the pyruvate dehydrogenase kinase isoform PDK1 at micromolar concentrations. It is not the unrelated PDPK1 kinase of the PI3K pathway.
2 Pyruvate oxidation and glycolytic dependence PDH activity ↑; glycolytic dependence ↓; oxidative metabolism altered Metabolic effects possible R–G Reduces metabolic flexibility PDK1 inhibition is expected to relieve PDH suppression. Direct metabolic responses may vary by tumor type and formulation.
3 CBL-b and survival-protein stability CBL-b-mediated degradation ↑; PDK1, PI3K, Akt, ULK1 and BCL-xL ↓ Insufficiently characterized G Destabilizes oncogenic and anti-apoptotic proteins Mechanistic interaction data were primarily obtained in AML cells and require confirmation in additional systems.
4 Mitochondrial apoptosis BAX activation ↑; caspase-3 ↑; cleaved PARP ↑; apoptosis ↑ Possible cytotoxicity at higher concentrations G Promotes intrinsic apoptotic death Associated with loss of anti-apoptotic signaling and mitochondrial BAX oligomerization.
5 BCL-2 family survival signaling BCL-2 ↓; BCL-xL ↓ Insufficiently characterized G Lowers the apoptotic threshold BAX and BAD responses may be model-dependent; the strongest evidence concerns reduced BCL-2 and BCL-xL.
6 PI3K Akt mTOR signaling PI3K ↓; Akt ↓; mTOR signaling ↓ Potential suppression G Reduces proliferation and survival signaling This effect may be partly secondary to PDK1-associated protein destabilization rather than direct inhibition of every pathway component.
7 Autophagy machinery ULK1 ↓; Beclin-1 ↓; LC3-II ↓; ATG5 ↓; ATG7 ↓; autophagy ↓ Insufficiently characterized G Suppresses cytoprotective autophagy Autophagy inhibition was demonstrated mainly in AML cells and may enhance apoptosis under metabolic stress.
8 Proliferation and clonogenic survival Proliferation ↓; colony formation ↓; viability ↓ Lower short-term effect reported in PBMCs, but selectivity is limited G Produces cytostatic and cytotoxic effects Subsequent analogue-development work characterized unmodified DAP as weakly potent and poorly selective.
9 Migration and invasive phenotype Migration ↓; colony formation ↓ Not established G Restrains aggressive tumor behavior Reported particularly in prostate-cancer studies; evidence remains preclinical.
10 EGFR inhibitor sensitization Response to erlotinib and gefitinib ↑; apoptosis ↑ Combination toxicity not adequately defined G Enhances EGFR-targeted therapy Synergy was reported in EGFR-mutant and experimentally gefitinib-resistant NSCLC models.
11 Docetaxel resistance Docetaxel-resistant cell viability ↓ Not established G Potential chemosensitization Stronger activity was observed with lactoferrin-encapsulated DAP than with free DAP.
12 ROS and DNA damage Uncertain Uncertain R–G Not established as a primary DAP mechanism General ROS production and DNA damage should not be presented as verified core mechanisms without compound-specific experimental evidence.
13 Clinical Translation Constraint Poor solubility; limited stability; weak selectivity; formulation-dependent activity Systemic safety and therapeutic index unknown G Restricts clinical development No human PK, dose-escalation, safety, or efficacy data are available. Nanoparticle delivery improves preclinical performance but is not clinically validated.

P: 0–30 min    R: 30 min–3 hr    G: >3 hr
ROS, Reactive Oxygen Species: Click to Expand ⟱
Source: HalifaxProj (inhibit)
Type:
Reactive oxygen species (ROS) are highly reactive molecules that contain oxygen and can lead to oxidative stress in cells. They play a dual role in cancer biology, acting as both promoters and suppressors of cancer.
ROS can cause oxidative damage to DNA, leading to mutations that may contribute to cancer initiation and progression. So normally you want to inhibit ROS to prevent cell mutations.
However excessive ROS can induce apoptosis (programmed cell death) in cancer cells, potentially limiting tumor growth. Chemotherapy typically raises ROS.
-mitochondria is the main source of reactive oxygen species (ROS) (and the ETC is heavily related)
ROS Homeostasis in Cancer Cells and Its Potential as a Therapeutic Target

"Reactive oxygen species (ROS) are two electron reduction products of oxygen, including superoxide anion, hydrogen peroxide, hydroxyl radical, lipid peroxides, protein peroxides and peroxides formed in nucleic acids 1. They are maintained in a dynamic balance by a series of reduction-oxidation (redox) reactions in biological systems and act as signaling molecules to drive cellular regulatory pathways."
"During different stages of cancer formation, abnormal ROS levels play paradoxical roles in cell growth and death 8. A physiological concentration of ROS that maintained in equilibrium is necessary for normal cell survival. Ectopic ROS accumulation promotes cell proliferation and consequently induces malignant transformation of normal cells by initiating pathological conversion of physiological signaling networks. Excessive ROS levels lead to cell death by damaging cellular components, including proteins, lipid bilayers, and chromosomes. Therefore, both scavenging abnormally elevated ROS to prevent early neoplasia and facilitating ROS production to specifically kill cancer cells are promising anticancer therapeutic strategies, in spite of their contradictoriness and complexity."
"ROS are the collection of derivatives of molecular oxygen that occur in biology, which can be categorized into two types, free radicals and non-radical species. The non-radical species are hydrogen peroxide (H 2O 2 ), organic hydroperoxides (ROOH), singlet molecular oxygen ( 1 O 2 ), electronically excited carbonyl, ozone (O3 ), hypochlorous acid (HOCl, and hypobromous acid HOBr). Free radical species are super-oxide anion radical (O 2•−), hydroxyl radical (•OH), peroxyl radical (ROO•) and alkoxyl radical (RO•) [130]. Any imbalance of ROS can lead to adverse effects. H2 O 2 and O 2 •− are the main redox signalling agents. The cellular concentration of H2 O 2 is about 10−8 M, which is almost a thousand times more than that of O2 •−".
"Radicals are molecules with an odd number of electrons in the outer shell [393,394]. A pair of radicals can be formed by breaking a chemical bond or electron transfer between two molecules."

Recent investigations have documented that polyphenols with good antioxidant activity may exhibit pro-oxidant activity in the presence of copper ions, which can induce apoptosis in various cancer cell lines but not in normal cells. "We have shown that such cell growth inhibition by polyphenols in cancer cells is reversed by copper-specific sequestering agent neocuproine to a significant extent whereas iron and zinc chelators are relatively ineffective, thus confirming the role of endogenous copper in the cytotoxic action of polyphenols against cancer cells. Therefore, this mechanism of mobilization of endogenous copper." > Ions could be one of the important mechanisms for the cytotoxic action of plant polyphenols against cancer cells and is possibly a common mechanism for all plant polyphenols. In fact, similar results obtained with four different polyphenolic compounds in this study, namely apigenin, luteolin, EGCG, and resveratrol, strengthen this idea.
Interestingly, the normal breast epithelial MCF10A cells have earlier been shown to possess no detectable copper as opposed to breast cancer cells [24], which may explain their resistance to polyphenols apigenin- and luteolin-induced growth inhibition as observed here (Fig. 1). We have earlier proposed [25] that this preferential cytotoxicity of plant polyphenols toward cancer cells is explained by the observation made several years earlier, which showed that copper levels in cancer cells are significantly elevated in various malignancies. Thus, because of higher intracellular copper levels in cancer cells, it may be predicted that the cytotoxic concentrations of polyphenols required would be lower in these cells as compared to normal cells."

Majority of ROS are produced as a by-product of oxidative phosphorylation, high levels of ROS are detected in almost all cancers.
-It is well established that during ER stress, cytosolic calcium released from the ER is taken up by the mitochondrion to stimulate ROS overgeneration and the release of cytochrome c, both of which lead to apoptosis.

Note: Products that may raise ROS can be found using this database, by:
Filtering on the target of ROS, and selecting the Effect Direction of ↑

Targets to raise ROS (to kill cancer cells):
• NADPH oxidases (NOX): NOX enzymes are involved in the production of ROS.
    -Targeting NOX enzymes can increase ROS levels and induce cancer cell death.
    -eNOX2 inhibition leads to a high NADH/NAD⁺ ratio which can lead to increased ROS
• Mitochondrial complex I: Inhibiting can increase ROS production
• P53: Activating p53 can increase ROS levels(by inducing the expression of pro-oxidant genes)
• Nrf2 inhibition: regulates the expression of antioxidant genes. Inhibiting Nrf2 can increase ROS levels
• Glutathione (GSH): an antioxidant. Depleting GSH can increase ROS levels
• Catalase: Catalase converts H2O2 into H2O+O. Inhibiting catalase can increase ROS levels
• SOD1: converts superoxide into hydrogen peroxide. Inhibiting SOD1 can increase ROS levels
• PI3K/AKT pathway: regulates cell survival and metabolism. Inhibiting can increase ROS levels
• HIF-1α inhibition: regulates genes involved in metabolism and angiogenesis. Inhibiting HIF-1α can increase ROS
• Glycolysis: Inhibiting glycolysis can increase ROS levels • Fatty acid oxidation: Cancer cells often rely on fatty acid oxidation for energy production.
-Inhibiting fatty acid oxidation can increase ROS levels
• ER stress: Endoplasmic reticulum (ER) stress can increase ROS levels
• Autophagy: process by which cells recycle damaged organelles and proteins.
-Inhibiting autophagy can increase ROS levels and induce cancer cell death.
• KEAP1/Nrf2 pathway: regulates the expression of antioxidant genes.
    -Inhibiting KEAP1 or activating Nrf2 can increase ROS levels and induce cancer cell death.
• DJ-1: regulates the expression of antioxidant genes. Inhibiting DJ-1 can increase ROS levels
• PARK2: regulates the expression of antioxidant genes. Inhibiting PARK2 can increase ROS levels
• SIRT1 inhibition:regulates the expression of antioxidant genes. Inhibiting SIRT1 can increase ROS levels
• AMPK activation: regulates energy metabolism and can increase ROS levels when activated.
• mTOR inhibition: regulates cell growth and metabolism. Inhibiting mTOR can increase ROS levels
• HSP90 inhibition: regulates protein folding and can increase ROS levels when inhibited.
• Proteasome: degrades damaged proteins. Inhibiting the proteasome can increase ROS levels
• Lipid peroxidation: a process by which lipids are oxidized, leading to the production of ROS.
    -Increasing lipid peroxidation can increase ROS levels
• Ferroptosis: form of cell death that is regulated by iron and lipid peroxidation.
    -Increasing ferroptosis can increase ROS levels
• Mitochondrial permeability transition pore (mPTP): regulates mitochondrial permeability.
    -Opening the mPTP can increase ROS levels
• BCL-2 family proteins: regulate apoptosis and can increase ROS levels when inhibited.
• Caspase-independent cell death: a form of cell death that is regulated by ROS.
    -Increasing caspase-independent cell death can increase ROS levels
• DNA damage response: regulates the repair of DNA damage. Increasing DNA damage can increase ROS
• Epigenetic regulation: process by which gene expression is regulated.
    -Increasing epigenetic regulation can increase ROS levels

-PKM2, but not PKM1, can be inhibited by direct oxidation of cysteine 358 as an adaptive response to increased intracellular reactive oxygen species (ROS)

ProOxidant Strategy:(inhibit the Mevalonate Pathway (likely will also inhibit GPx)
-HydroxyCitrate (HCA) found as supplement online and typically used in a dose of about 1.5g/day or more
-Atorvastatin typically 40-80mg/day, -Dipyridamole typically 200mg 2x/day Combined effect research
-Lycopene typically 100mg/day range (note debatable as it mainly lowers NRF2)

Dual Role of Reactive Oxygen Species and their Application in Cancer Therapy 
ROS-Inducing Interventions in Cancer — Canonical + Mechanistic Reference
-generated from AI and Cancer database
ROS rating:  +++ strong | ++ moderate | + weak | ± mixed | 0 none
NRF2:        ↓ suppressed | ↑ activated | ± mixed | 0 none
Conditions:  [D] dose  [Fe] metal  [M] metabolic  [O₂] oxygen
             [L] light [F] formulation [T] tumor-type [C] combination




Item ROS↑ NRF2 Condition Mechanism Class Remarks 



ROS">Piperlongumine
+++ ↓ [D][T] ROS-dominant






ROS">Shikonin
+++↓/±[D][T]ROS-dominant






ROS">Vitamin K3 (menadione)
+++↓[D]ROS-dominant






ROS">Copper (ionic / nano)
+++↓[Fe][F]ROS-dominant






ROS">Sodium Selenite
+++↓[D]ROS-dominant






ROS">Juglone
+++↓[D]ROS-dominant






ROS">Auranofin
+++↓[D]ROS-dominant






ROS">Photodynamic Therapy (PDT)
+++0[L][O₂]ROS-dominant






ROS">Radiotherapy / Radiation
+++0[O₂]ROS-dominant






ROS">Doxorubicin
+++↓[D]ROS-dominant






ROS">Cisplatin
++↓[D][T]ROS-dominant






ROS">Salinomycin
++↓[D][T]ROS-dominant






ROS">Artemisinin / DHA
++↓[Fe][T]ROS-dominant






ROS">Sulfasalazine
++↓[C][T]ROS-dominant






ROS">FMD / fasting
++↓[M][C][O₂]ROS-dominant






ROS">Vitamin C (pharmacologic)
++↓[Fe][D]ROS-dominant






ROS">Silver nanoparticles
++±[F][D]ROS-dominant






ROS">Gambogic acid
++↓[D][T]ROS-dominant






ROS">Parthenolide
++↓[D][T]ROS-dominant






ROS">Plumbagin
++↓[D]ROS-dominant






ROS">Allicin
++↓[D]ROS-dominant






ROS">Ashwagandha (Withaferin A)
++↓[D][T]ROS-dominant






ROS">Berberine
++↓[D][M]ROS-dominant






ROS">PEITC
++↓[D][C]ROS-dominant






ROS">Methionine restriction
+↓[M][C][T]ROS-secondary






ROS">DCA
+±[M][T]ROS-secondary






ROS">Capsaicin
+±[D][T]ROS-secondary






ROS">Galloflavin
+0[D]ROS-secondary






ROS">Piperine
+±[D][F]ROS-secondary






ROS">Propyl gallate
+↓[D]ROS-secondary






ROS">Scoulerine
+?[D][T]ROS-secondary






ROS">Thymoquinone
±±[D][T]Dual redox






ROS">Emodin
±±[D][T]Dual redox






ROS">Alpha-lipoic acid (ALA)
±↑[D][M]NRF2-dominant






ROS">Curcumin
±↑/↓[D][F]NRF2-dominant






ROS">EGCG
±↑/↓[D][O₂]NRF2-dominant






ROS">Quercetin
±↑/↓[D][Fe]NRF2-dominant






ROS">Resveratrol
±↑[D][M]NRF2-dominant






ROS">Sulforaphane
±↑↑[D]NRF2-dominant






ROS">Lycopene
0↑—Antioxidant






ROS">Rosmarinic acid
0↑—Antioxidant






ROS">Citrate
00—Neutral


Scientific Papers found: Click to Expand⟱
6685- DAP,    Anticancer effects of some novel dichloroacetophenones through the inhibition of pyruvate dehydrogenase kinase 1
- in-vitro, Lung, H1975
PDKs↓, selectivity∅, MMP↓, Apoptosis↑, lactateProd↓, ROS↑,

Showing Research Papers: 1 to 1 of 1

* indicates research on normal cells as opposed to diseased cells
Total Research Paper Matches: 1

Pathway results for Effect on Cancer / Diseased Cells:


Redox & Oxidative Stress(tgid=1) ⓘ

ROS↑, 1,  

Mitochondria & Bioenergetics(tgid=3) ⓘ

MMP↓, 1,  

Core Metabolism/Glycolysis(tgid=4) ⓘ

lactateProd↓, 1,   PDKs↓, 1,  

Cell Death(tgid=5) ⓘ

Apoptosis↑, 1,  

Drug Metabolism & Resistance(tgid=21) ⓘ

selectivity∅, 1,  
Total Targets: 6

Pathway results for Effect on Normal Cells:


Total Targets: 0

Scientific Paper Hit Count for: ROS, Reactive Oxygen Species
Query results interpretion may depend on "conditions" listed in the research papers.
Such Conditions may include : 
  -low or high Dose
  -format for product, such as nano of lipid formations
  -different cell line effects
  -synergies with other products 
  -if effect was for normal or cancerous cells

Filter Conditions: Pro/AntiFlg:%  IllCat:%  CanType:%  Cells:%  prod#:289  Target#:275  State#:%  Dir#:2
  wNotes=0  sortOrder:rid,rpid

 

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