Arecaidine-propargyl ester
Arecaidine-propargyl ester is a selective M2 muscarinic receptor agonist with blood-brain barrier permeability, with a pKi of 5.91 for hm1, 7.06 for hm2, 6.07 for hm3, 6.01 for hm4, and 6.03 for hm5. Arecaidine-propargyl ester stimulates central and peripheral muscarinic receptors. Arecaidine-propargyl ester increases intracellular ROS, induces DNA damage and Apoptosis, and upregulates the expression of MnSOD and SIRT1. Arecaidine-propargyl ester reduces sympathetic nerve outflow, induces dose-dependent hypotension, and triggers negative chronotropic effects at high peripheral doses. Arecaidine-propargyl ester can be used in research related to Alzheimer's disease and glioblastoma.
For research use only. We do not sell to patients.
- CAS No.: 35516-99-5
- Formula: C10H13NO2
- Molecular Weight:179.22
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Storage:
Please store the product under the recommended conditions in the Certificate of Analysis.
All DNA/RNA Synthesis Isoforms
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Biological Activity
Description
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mAChR1 5.91 (pKi) |
mAChR2 7.06 (pKi) |
mAChR3 6.07 (pKi) |
mAChR4 6.01 (pKi) |
mAChR5 6.03 (pKi) |
SIRT1 |
In Vitro
Arecaidine-propargyl ester (0.01-1000 mM; 120 min) binds to cloned hm1, hm2, hm3, hm4, and hm5 muscarinic receptors expressed in CHO cells, with pKi values ranging from 5.91 to 7.06[2].
Arecaidine-propargyl ester (25-100 μM; 2 h) induces significant intracellular ROS production in glioblastoma cell lines U251MG and U87MG, and this effect is completely blocked by the ROS scavenger NAC[3].
Arecaidine-propargyl ester (100 μM; 72 h) induces significant apoptosis and reduces cell numbers in human glioblastoma cell lines U251MG and U87MG, with a stronger apoptotic effect on U251MG cells. Both of these effects are abrogated by the ROS scavenger NAC[3].
Arecaidine-propargyl ester (100 μM; 24-48 h) significantly upregulates the expression of SIRT1 protein in glioblastoma U251MG and U87MG cells in a time-dependent manner in vitro[3].
Arecaidine-propargyl ester (100 μM; 24-48 h) significantly upregulates the expression of MnSOD protein in U251MG and U87MG glioblastoma cells in a time-dependent manner[3].
Arecaidine-propargyl ester (100 μM; 24-48 h) significantly upregulates the mRNA expression level of Gadd45α in glioblastoma cell lines U251MG and U87MG[3].
Arecaidine-propargyl ester (100 μM; 24 h) increases total glutathione levels by 50% in U251MG glioblastoma cells, whereas no significant metabolic changes are detected in U87MG cells[3].
Arecaidine-propargyl ester (100 μM; 24-48 h) induces DNA double-strand breaks (detected by γ-H2AX positive rate) in glioblastoma cell lines U251MG and U87MG, and this effect depends on the activation of M2 muscarinic receptors in U251MG cells[3].
Arecaidine-propargyl ester (50-100 μM; 24 h) induces significant chromosomal aberrations in glioblastoma U251MG and U87MG cells; among them, U251MG cells exhibit higher sensitivity at 50 μM, and only U251MG cells show a significant increase in sister chromatid exchange levels at 50 μM[3].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:U251MG, U87MG human glioblastoma cell lines
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Concentration:100 μM
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Incubation Time:72 h
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Result:Increased the apoptotic index in U87MG cells and U251MG cells.
Significantly reduced the apoptotic index in both cell lines, returning levels near control values when co-treated with 5 μM NAC.
Caused a significant decrease in cell number in both lines, an effect reversed by co-treatment with NAC.
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Cell Line:U251MG, U87MG human glioblastoma cell lines
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Concentration:100 μM
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Incubation Time:24 h, 30 h, 48 h
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Result:Caused a significant increase in Gadd45α mRNA expression in both cell lines.
Increased Gadd45α/18S ratios in U87MG cells.
Increased Gadd45α/18S ratios in U251MG cells.
In Vivo
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:either sex, weight 2-4 kg (anaesthetized for cardiovascular function monitoring)[1]
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Dosage:1.0 × 10-9 moles/kg (vertebral artery); 1.4 × 10-9 moles/kg (femoral vein)
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Administration:left vertebral artery; femoral vein
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Result:Caused dose-dependent reduction in mean arterial pressure (intravenous), which was blocked by intravenous methylatropine (300 μg/kg) but not centrally administered atropine (30 μg/kg) or D-benzethimide (10 μg/kg).
Caused dose-dependent reduction in mean arterial pressure (vertebral artery), which was not blocked by intravenous methylatropine (300 μg/kg) but was blocked by vertebral artery-administered atropine (50 μg/kg) or D-benzethimide (10 μg/kg).
Did not alter heart rate with central administration; only relatively high intravenous doses caused a decrease in cardiac frequency.
Exhibited ED-25 of 1.0 × 10-9 moles/kg via vertebral artery and 1.4 × 10-9 moles/kg via femoral vein.
Chemical Information
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CAS No. 35516-99-5
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Molecular Weight 179.22
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Formula C10H13NO2
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SMILES
O=C(C1=CCCN(C1)C)OCC#C
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Please store the product under the recommended conditions in the Certificate of Analysis.
Protocols
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RNA extraction experimental
By lysing cells, releasing RNA, and removing impurities such as proteins and DNA, high-purity RNA products are finally obtained. The commonly used traditional method is the guanidine isothiocyanate/phenol/chloroform method (Trizol), which is suitable for a variety of animal materials including animal tissues, microorganisms, cultured cells, etc., and most plant materials.
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Apoptosis
Apoptosis, also called programmed cell death, is generally characterized by distinct morphological characteristics.
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TUNEL staining for apoptotic DNA fragmentation
TUNEL staining detects DNA strand breaks by using terminal deoxynucleotidyl transferase to add labeled nucleotides to exposed 3′-OH DNA termini, generating either microscopic staining in fixed cells or tissue sections, or fluorescence/cytometric signal in cell suspensions. TUNEL positivity reflects DNA fragmentation but should not be interpreted alone as definitive apoptosis, because TUNEL can also label necrotic, autolytic, mechanically damaged, or DNA-repair-associated DNA breaks.
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Annexin V plus membrane-impermeant dye apoptosis staining
Annexin V-based apoptosis assays rely on the detection of phosphatidylserine (PS) externalization from the inner leaflet of the plasma membrane to the outer leaflet, an early biochemical hallmark of apoptosis. Fluorescently labeled Annexin V binds PS in a calcium-dependent manner, enabling identification of early apoptotic cells by flow cytometry or fluorescence microscopy. When combined with a membrane-impermeant DNA-binding dye (e. g. , propidium iodide), this approach allows discrimination between viable (Annexin V−/dye−), early apoptotic (Annexin V+/dye−), and late apoptotic or necrotic (Annexin V+/dye+) cell populations by assessing membrane integrity and PS exposure.
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ROS/oxidative-stress fluorescent staining
ROS/oxidative-stress fluorescent staining uses cell-permeant fluorogenic probes that become fluorescent after oxidation inside cells or tissues; commonly used examples include DCFH-DA/DCFDA for broad cellular oxidant detection, DHE for superoxide-related signal detection, MitoSOX for mitochondrial superoxide-related signal detection, and CellROX probes for oxidative-stress-associated fluorescence readouts. The assay detects probe oxidation rather than a single ROS species unless the probe and analysis method have been chemically validated for that species. DCFH-DA enters cells, is deacetylated by intracellular esterases to DCFH, and produces fluorescent DCF after oxidation, so the readout is used as an operational measure of total cellular oxidative stress rather than a species-specific ROS measurement. DHE and MitoSOX can report superoxide-related oxidation, but red fluorescence alone can include non-specific ethidium-like oxidation products; HPLC or optimized spectral approaches are
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Transepithelial/transendothelial electrical resistance assay
TEER measures electrical resistance across epithelial or endothelial monolayers cultured on permeable supports, and the readout reflects ionic conductance through the cell barrier, especially the paracellular pathway regulated by junctional integrity. TEER can be measured without destroying the monolayer and is commonly used before or during transport, permeability, barrier-disruption, and barrier-maturation experiments. TEER values are influenced by biological maturation and technical conditions; reported factors include temperature, medium formulation, passage number, electrode geometry, membrane properties, and junctional length during early monolayer maturation. Therefore, TEER should be interpreted with blank-insert subtraction, area normalization, repeated readings, and, when possible, orthogonal barrier readouts such as FITC-dextran flux or tight-junction staining.
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Apoptosis Solutions
Apoptosis is a regulated, generally non-lytic cell-death pathway that removes unwanted, damaged, infected, or abnormal cells through coordinated morphological changes, caspase activation, DNA fragmentation, and membrane remodeling. The intrinsic apoptosis pathway is controlled mainly by mitochondrial outer membrane permeabilization, BCL-2 family proteins, cytochrome c release, apoptosome formation, caspase-9 activation, and downstream executioner caspase-3/7 activation. The extrinsic apoptosis pathway is initiated by death receptors such as Fas, TNFR, and TRAIL receptors, which recruit adaptor proteins and activate caspase-8 before engaging executioner caspases or mitochondrial amplification through BID cleavage. Apoptosis is linked to many phenotypes, including cancer cell killing, tissue homeostasis, immune regulation, neurodegeneration, infection response, and treatment-induced cytotoxicity; unresolved questions include how apoptosis interacts with necroptosis, pyroptosis, ferroptos
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Genotoxicity/Mutagenicity Study
The bacterial reverse mutation assay detects point mutations that restore amino-acid prototrophy in auxotrophic Salmonella typhimurium or Escherichia coli tester strains; after exposure to a test article, mutagenic activity is read out as an increased number of revertant colonies on minimal agar compared with the vehicle control. The assay uses tester strains with different mutation targets so that base-substitution and frameshift mutagens can be detected, and testing is performed with and without exogenous mammalian metabolic activation because some chemicals require biotransformation to become mutagenic.
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Alzheimer’s Disease Modeling
Alzheimer’s Disease (AD) is a neurodegenerative disorder characterized by a progressive decline in cognitive functions and loss of specific types of neurons and synapses. Alzheimer's symptoms can be simulated in mice by injecting drugs (such as Aβ) or genetically modified.
Purity & Documentation
References
[1]. Porsius AJ, et al. Central action of some cholinergic drugs (arecaidine esters) and nicotine on blood pressure and heart rate of cats. Prog Brain Res. 1977;47:131-5. [Content Brief]
[2]. Scapecchi S, et al. Highly chiral muscarinic ligands: the discovery of (2S,2'R,3'S,5'R)-1-methyl-2-(2-methyl-1,3-oxathiolan-5-yl)pyrrolidine 3-sulfoxide methyl iodide, a potent, functionally selective, M2 partial agonist. J Med Chem. 2006 Mar 23;49(6):1925-31. [Content Brief]
[3]. Di Bari M, et al. Cytotoxic and genotoxic effects mediated by M2 muscarinic receptor activation in human glioblastoma cells. Neurochem Int. 2015;90:261-270. [Content Brief]
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)