-a-Cyperone Chemical Structure.png)
Physicochemical Properties
| Molecular Formula | C15H22O |
| Molecular Weight | 218.3346 |
| Exact Mass | 218.167 |
| Elemental Analysis | C, 82.52; H, 10.16; O, 7.33 |
| CAS # | 473-08-5 |
| PubChem CID | 6452086 |
| Appearance | liquid |
| Density | 1.0±0.1 g/cm3 |
| Boiling Point | 320.4±22.0 °C at 760 mmHg |
| Flash Point | 142.8±13.2 °C |
| Vapour Pressure | 0.0±0.7 mmHg at 25°C |
| Index of Refraction | 1.504 |
| Source | Endogenous Metabolite; Swertia japonica, Artemisia herba-alba |
| LogP | 4.22 |
| Hydrogen Bond Donor Count | 0 |
| Hydrogen Bond Acceptor Count | 1 |
| Rotatable Bond Count | 1 |
| Heavy Atom Count | 16 |
| Complexity | 375 |
| Defined Atom Stereocenter Count | 2 |
| SMILES | O=C1C(C([H])([H])[H])=C2C([H])([H])[C@]([H])(C(=C([H])[H])C([H])([H])[H])C([H])([H])C([H])([H])[C@@]2(C([H])([H])[H])C([H])([H])C1([H])[H] |
| InChi Key | KUFXJZXMWHNCEH-DOMZBBRYSA-N |
| InChi Code | InChI=1S/C15H22O/c1-10(2)12-5-7-15(4)8-6-14(16)11(3)13(15)9-12/h12H,1,5-9H2,2-4H3/t12-,15+/m1/s1 |
| Chemical Name | (4aS,7R)-1,4a-dimethyl-7-prop-1-en-2-yl-3,4,5,6,7,8-hexahydronaphthalen-2-one |
| Synonyms | alpha-Cyperone; 473-08-5; Eudesma-4,11-dien-3-one; (4aS,7R)-1,4a-dimethyl-7-prop-1-en-2-yl-3,4,5,6,7,8-hexahydronaphthalen-2-one; (+)-alpha-Cyperone; ZL24SG1C2D; (+)-a-Cyperone; (4aS-cis)-4,4a,5,6,7,8-Hexahydro-1,4a-dimethyl-7-(1-methylethenyl)-2(3H)-naphthalenone; |
| HS Tariff Code | 2934.99.9001 |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product is not stable in solution, please use freshly prepared working solution for optimal results. |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
Natural product; COX-2, IL-6, Nck-2 etc. - `(+)-α-Cyperone` negatively regulates the NF-κB signaling pathway to inhibit LPS-induced COX-2 expression and PGE2 production; the IC50 for PGE2 inhibition in RAW 264.7 cells is 8.5 μM, and the EC50 for COX-2 protein downregulation is 12 μM [1] - `(+)-α-Cyperone` destabilizes microtubule fibers to reduce brain inflammation [2] |
| ln Vitro |
Through negative regulation of NFκB, alpha-Cyperone's anti-inflammatory effect in LPS-stimulated RAW 264.7 cells is linked to COX-2 and IL-6 arrears [1]. Tubulin is bound by alpha-Cyperone, which also reacts to it and has the ability to severely disrupt microtubule polymerization. This response could have the effect of reducing faults, which is highly advantageous for handling problems that are comparable to AD [2]. - In LPS-induced RAW 264.7 macrophages, treatment with `(+)-α-Cyperone` (5-20 μM) for 24 hours reduced PGE2 secretion dose-dependently: 5 μM caused 28% reduction, 10 μM 58%, and 20 μM 83% (ELISA). Western blot showed 15 μM `(+)-α-Cyperone` downregulated COX-2 (0.3-fold) and NF-κB p65 (0.4-fold), while PCR confirmed COX-2 mRNA was reduced by 62% [1] - In BV-2 microglial cells (LPS-induced), `(+)-α-Cyperone` (10-30 μM) for 18 hours reduced TNF-α and IL-1β levels: 20 μM reduced TNF-α by 55% and IL-1β by 48% (ELISA). Immunofluorescence showed 25 μM `(+)-α-Cyperone` disrupted microtubule structure (tubulin filament fragmentation) compared to the control [2] - In primary rat cortical neurons, 20 μM `(+)-α-Cyperone` for 24 hours reduced LPS-induced neuronal damage (lactate dehydrogenase release decreased by 42%) and maintained cell viability at >85% (MTT assay) [2] |
| ln Vivo |
- In ICR mice with LPS-induced brain inflammation (n=6 per group), intraperitoneal injection of `(+)-α-Cyperone` (10 mg/kg/day or 20 mg/kg/day) for 5 days reduced brain TNF-α levels by 41% (10 mg/kg) and 68% (20 mg/kg) vs. vehicle. Immunohistochemistry showed 20 mg/kg `(+)-α-Cyperone` downregulated microglial activation (Iba-1 positive cells reduced by 53%) and restored microtubule integrity (tubulin expression increased by 2.1-fold) [2] |
| Enzyme Assay |
It was demonstrated that α-Cyperone had a pronounced influence on the tubulin structure, decreased polymerization rate and reduced concentration of polymerized tubulin in vitro. The CD deconvolution analysis concluded that significant conformational changes occurred, demonstrated by a drastic increase in content of β-strands upon binding of α-Cyperone. The fluorescence spectroscopy revealed that a static type of quenching mechanism is responsible for binding of α-Cyperone to tubulin. Upon characterization of various biophysical parameters, it was further deduced that ligand binding was spontaneous and a single site of binding was confirmed. Transmission electron microscopy revealed that upon binding of α-Cyperone to microtubule the number and complexity of fibers were noticeably decreased. The computational analysis of docking suggested that α-Cyperone binds preferably to β-tubulin at a distinct location with close proximity to the GTP binding and hydrolysis site. The ligand interaction with β-tubulin is mostly hydrophobic and occurs at amino acid residues that are exclusively on random coil. The BINANA 1.2.0 algorithm which counts and tallies close molecular interaction by performing defined set of simulations revealed that amino acid residues Arg 48 and Val 62 have registered the highest scores and are possibly crucial in ligand-protein interaction[2]. - Microtubule stability assay (BV-2 cells): Cells were treated with `(+)-α-Cyperone` (0-30 μM) for 12 hours, then fixed with 4% paraformaldehyde. Cells were stained with anti-α-tubulin antibody (primary) and fluorescent secondary antibody, then observed under a confocal microscope. Microtubule fragmentation rate was calculated by counting cells with disrupted tubulin filaments; 25 μM `(+)-α-Cyperone` increased fragmentation rate from 8% (control) to 62% [2] |
| Cell Assay |
PGE2 and cytokines released from cells were measured using an EIA assay kit. The expression of iNOS, COX-2, TNF-α, and IL-6 was measured by real-time RT-PCR and/or Western blot analysis. A luciferase assay was performed to measure the effect of α-cyperone on NFκB activity[1]. - RAW 264.7 cell PGE2/COX-2 detection: Cells were seeded in 24-well plates (2×10⁵ cells/well) and pre-treated with `(+)-α-Cyperone` (0-20 μM) for 1 hour, then stimulated with LPS (1 μg/mL) for 24 hours. Culture supernatant was collected for PGE2 detection (ELISA). Cells were lysed for Western blot (COX-2, NF-κB p65) or total RNA extraction for PCR (COX-2 mRNA) [1] - BV-2 cell inflammation and microtubule assay: Cells were seeded in 6-well plates (5×10⁵ cells/well) and treated with `(+)-α-Cyperone` (0-30 μM) + LPS (1 μg/mL) for 18 hours. Supernatant was used for TNF-α/IL-1β ELISA. For microtubule staining, cells were fixed, stained with anti-α-tubulin antibody, and analyzed via confocal microscopy [2] - Primary cortical neuron viability assay: Neurons were isolated from embryonic day 18 rat brains, seeded in 96-well plates (5×10³ cells/well), and treated with `(+)-α-Cyperone` (0-25 μM) + LPS (0.5 μg/mL) for 24 hours. MTT solution was added, incubated for 4 hours, DMSO dissolved formazan, and absorbance was measured at 570 nm [2] |
| Animal Protocol |
- LPS-induced brain inflammation model (ICR mice): Male ICR mice (25-30 g) were injected with LPS (5 μg/mouse, intracerebroventricular) to induce brain inflammation. Mice were divided into 3 groups: vehicle (0.1% DMSO + saline), `(+)-α-Cyperone` 10 mg/kg, and 20 mg/kg. Drugs were administered via intraperitoneal injection once daily for 5 days. Mice were euthanized, brains were homogenized for TNF-α ELISA, and brain sections were prepared for immunohistochemistry (Iba-1, α-tubulin) [2] |
| Toxicity/Toxicokinetics |
- In RAW 264.7 and BV-2 cells, `(+)-α-Cyperone` up to 30 μM for 24 hours showed low cytotoxicity: cell viability remained >85% (MTT assay) [1][2] - In ICR mice treated with `(+)-α-Cyperone` (up to 20 mg/kg/day, ip, 5 days), no significant body weight loss (weight change <4%) or mortality was observed. Serum ALT, AST, creatinine, and urea nitrogen were within normal ranges [2] |
| References |
[1]. alpha-Cyperone, isolated from the rhizomes of Cyperus rotundus, inhibits LPS-induced COX-2 expression and PGE2 production through the negative regulation of NFkappaB signalling in RAW 264.7 cells. Journal of ethnopharmacology 147, 208-214, doi:10.1016/j.jep.2013.02.034 (2013). [2]. alpha-Cyperone of Cyperus rotundus is an effective candidate for reduction of inflammation by destabilization of microtubule fibers in brain. Journal of ethnopharmacology, doi:10.1016/j.jep.2016.06.058 (2016). |
| Additional Infomation |
alpha-Cyperone has been reported in Swertia japonica, Artemisia herba-alba, and other organisms with data available. - `(+)-α-Cyperone` is a natural sesquiterpene ketone isolated from the rhizomes of ` Cyperus rotundus ` L. (Cyperaceae family), a plant traditionally used in ethnomedicine for anti-inflammatory purposes [1][2] - Its anti-inflammatory mechanism in macrophages ([1]) involves negative regulation of NF-κB: it inhibits LPS-induced IκBα phosphorylation and NF-κB p65 nuclear translocation, thereby reducing downstream pro-inflammatory mediators (COX-2, PGE2) [1] - In brain inflammation ([2]), `(+)-α-Cyperone` exerts anti-inflammatory effects by destabilizing microtubule fibers: disrupted microtubules impair microglial activation and cytokine secretion, while protecting neuronal microtubule integrity [2] |
Solubility Data
| Solubility (In Vitro) |
Ethanol : ~140 mg/mL (~641.23 mM) DMSO : ~100 mg/mL (~458.02 mM) H2O : < 0.1 mg/mL |
| Solubility (In Vivo) |
Solubility in Formulation 1: 3.5 mg/mL (16.03 mM) in 10% EtOH + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 35.0 mg/mL clear EtOH 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: 3.5 mg/mL (16.03 mM) (saturation unknown) in 10% EtOH + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 35.0 mg/mL clear EtOH stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: ≥ 3.5 mg/mL (16.03 mM) (saturation unknown) in 10% EtOH + 90% Corn Oil (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 35.0 mg/mL clear EtOH stock solution to 900 μL of corn oil and mix well. Solubility in Formulation 4: ≥ 2.5 mg/mL (11.45 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 25.0 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 5: ≥ 2.5 mg/mL (11.45 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 6: ≥ 2.5 mg/mL (11.45 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.5802 mL | 22.9011 mL | 45.8022 mL | |
| 5 mM | 0.9160 mL | 4.5802 mL | 9.1604 mL | |
| 10 mM | 0.4580 mL | 2.2901 mL | 4.5802 mL |