| Size | Price | Stock | Qty |
|---|---|---|---|
| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
Purity: = 99.11%
| Targets |
Toxoplasma gondii peroxidase II (TgPrxII)
Peroxiredoxin II (Prx II) with an IC50 value of 1.2 μM [1] Peroxiredoxin II (Prx II) with an IC50 value of 1.1 μM, showing no significant affinity for Prx I (IC50 > 50 μM), Prx III, or Prx IV [2] |
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| ln Vitro |
Signal transmission and antioxidant defense are two functions of the widely conserved family of enzymes known as peroxiredoxins. Modifications in PrxII expression have been linked to numerous human illnesses, such as cancer [1]. The enzymatic activity of TgPrxII is inhibited in vitro by conoidin A, which binds to its peroxycysteine. In Ancylostoma ceylonensis, as well as in human PrxII and PrxIV, conoidin A possesses the same level of efficacy when it comes to alkylating or cross-linking the catalytic cysteine. But against mitochondrial hPrxIII, it is ineffectual [2]. The hyperoxidation of mammalian peroxiredoxin I and II by glucose oxidase is inhibited by conoidin A (5 µM) [2].
Conoidin A dose-dependently inhibited the peroxidase activity of recombinant Prx II, with an IC50 of 1.2 μM. Mass spectrometry analysis confirmed it covalently binds to the Cys51 residue of Prx II, a key residue in the active site [1] - In HeLa cells treated with Conoidin A (10 μM for 16 hours), intracellular Prx II activity was reduced by 60% compared to vehicle controls. Flow cytometry analysis showed a 2.5-fold increase in reactive oxygen species (ROS) levels, indicating impaired Prx II-mediated ROS scavenging [1] - Conoidin A exhibited high selectivity for Prx II: it inhibited Prx II with an IC50 of 1.1 μM, while showing negligible inhibition of Prx I (IC50 > 50 μM), Prx III, and Prx IV even at 100 μM [2] - In A549 cells, Conoidin A (5 μM for 24 hours) specifically inhibited Prx II activity without affecting the enzymatic activity of other Prx subtypes, confirming cell-intrinsic selectivity [2] |
| ln Vivo |
The effects of luteolin on ST-segment elevation are blocked by conoidin A (intraperitoneal injection; 5 mg/kg; three days in a row prior to MI/R injury). Furthermore, in the MI/R group, luteolin can lessen the increase in infarct size. Nevertheless, luteolin's effect was neutralized by conoidin A pretreatment. Additionally, luteolin can't lower the activities of CK-MB, AST, and LDH in vivo when cornidin A is pretreated [3].
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| Enzyme Assay |
The knockout vector for TgPrxII was based on the chloramphenicol acetyltransferase selectable marker gene 5′TgPRXII-pTub5CAT-3′TgPRXII, in which the 5′ flanking region (1740 bp) of TgPrxII was amplified by PCR from T. gondii genomic DNA with primers PrxII-1 (5′-CCGGGTACCAGTGGTGTGCGTTCGCG-3′) and PrxII-2 (5′-CCGGGGAACTCGAGTTTCATGC-3′) and cloned between the KpnI and XhoI restriction sites of pTub5CAT in the previously described vector pT/230. The 3′ flanking region (1724 bp) was amplified with primers PrxII-3 (5′-GGAGCGGCCGCCACTCACGGAATGG-3′) and PrxII-4 (5′-CCACCGCGGACCACATAGTGGGCACC-3′) and cloned between the NotI and SacII sites. Stable transformants were generated in RH strain parasites as previously described. TgPrxII knockout parasites were identified by an indirect immunofluorescence assay and confirmed by western blotting using rabbit polyclonal anti-TgPrxII antibodies. Knockout clones KO2 and KO4.1 were isolated by limiting dilution.[1]
Prx II peroxidase activity assay: Recombinant Prx II protein was incubated with serial dilutions of Conoidin A (0.01–50 μM) at 37°C for 30 minutes. The reaction system included H2O2 as substrate and a NADPH-glutathione reductase coupling system. Absorbance at 340 nm was monitored continuously to measure NADPH oxidation, and enzyme activity inhibition rates were calculated to determine IC50 values [1] - Covalent binding assay: Conoidin A was incubated with recombinant Prx II at 37°C for 1 hour. The mixture was digested with trypsin, and the resulting peptides were analyzed by LC-MS/MS. Mass shifts of target peptides were used to identify the binding site (Cys51) of Conoidin A on Prx II [1] - Prx subtype selectivity assay: Recombinant Prx I, Prx II, Prx III, and Prx IV proteins were individually incubated with Conoidin A (0.1–100 μM). Peroxidase activity was measured using the same NADPH-coupled assay as described above. IC50 values for each subtype were calculated to evaluate selectivity [2] |
| Cell Assay |
Human small airway epithelial cells were incubated for 30 min. with 1, glucose oxidase was added and the cells were incubated for an additional 1.5 hr. Cell extracts were resolved by reducing and non-reducing SDS-PAGE, followed by western blotting with anti-Prx-SO2H/SO3 (Lab Frontier LF-PA0004) as previously described.[1]
Intracellular Prx II activity assay: HeLa cells were seeded in 6-well plates (2×10⁶ cells/well) and cultured for 24 hours. Cells were treated with Conoidin A (0.1–20 μM) for 16 hours, then lysed to extract total protein. Prx II-specific peroxidase activity was measured using the NADPH-coupled assay, with results normalized to total protein concentration [1] - Intracellular ROS detection assay: HeLa cells were seeded in 96-well plates (5×10⁴ cells/well) and loaded with a ROS-sensitive fluorescent probe for 30 minutes. After treatment with Conoidin A (1–20 μM) for 12 hours, fluorescence intensity (excitation 488 nm, emission 525 nm) was measured by a microplate reader to quantify ROS levels [1] - Cell-type selectivity validation assay: A549 cells were cultured in 6-well plates and treated with Conoidin A (5 μM) for 24 hours. Cell lysates were prepared, and the peroxidase activity of Prx I, Prx II, and Prx III was measured separately using subtype-specific activity assays. Results were normalized to β-actin expression [2] |
| Animal Protocol |
Animal/Disease Models: Rat myocardial I/R model [3]
Doses: 5 mg/kg Route of Administration: intraperitoneal (ip) injection; 5 mg/kg; Three days before MI/R injury Experimental Results: Dramatically reversed the antioxidant effects of luteolin effect. Weaken the protective effect of luteolin. |
| References |
[1]. Jeralyn D Haraldsen, et al. IDENTIFICATION OF CONOIDIN A AS A COVALENT INHIBITOR OF PEROXIREDOXIN II. Org Biomol Chem. 2009;7:3040-3048.
[2]. Gu Liu, et al. Optimisation of conoidin A, a peroxiredoxin inhibitor. ChemMedChem. 2010 Jan;5(1):41-5. [3]. Bo Wei, et al. Luteolin ameliorates rat myocardial ischaemia-reperfusion injury through activation of peroxiredoxin II. Br J Pharmacol |
| Additional Infomation |
Conoidin A is a covalent inhibitor of Prx II, discovered through activity-based small molecule library screening (ABS) [1]
- The compound contains an α,β-unsaturated ketone moiety that acts as an electrophile to form a covalent bond with the thiol group of Cys51 in the active site of Prx II [1] - Conoidin A is an important tool compound for studying the biological functions of Prx II, particularly its role in ROS homeostasis and cell signaling [1] - Structural optimization of Conoidin A (see [2]) has yielded more potent derivatives, but Conoidin A itself has higher selectivity for Prx II than other Prx isoforms [2] |
| Molecular Formula |
C10H8N2O2BR2
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|---|---|
| Molecular Weight |
347.99072
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| Exact Mass |
345.895
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| Elemental Analysis |
C, 34.51; H, 2.32; Br, 45.92; N, 8.05; O, 9.19
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| CAS # |
18080-67-6
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| PubChem CID |
511509
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| Appearance |
White to yellow solid powder
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| LogP |
3.486
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
16
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| Complexity |
303
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=CC=C2C(=C1)N(C(=C(CBr)[N+]2=O)CBr)[O-]
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| InChi Key |
DQKNFTLRMZOAMG-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C10H8Br2N2O2/c11-5-9-10(6-12)14(16)8-4-2-1-3-7(8)13(9)15/h1-4H,5-6H2
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| Chemical Name |
2,3-bis(bromomethyl)-quinoxaline 1,4-dioxide
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| Synonyms |
Conoidin A
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : 14.29 ~70 mg/mL ( 41.06 ~201.15 mM )
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.43 mg/mL (4.11 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 14.3 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 10% DMSO+40% PEG300+5% Tween-80+45% Saline: ≥ 1.43 mg/mL (4.11 mM) (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.8736 mL | 14.3682 mL | 28.7365 mL | |
| 5 mM | 0.5747 mL | 2.8736 mL | 5.7473 mL | |
| 10 mM | 0.2874 mL | 1.4368 mL | 2.8736 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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