| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
Purity: =97.1%
| Targets |
NMDA Receptor
Withanone is a multi‑target compound. Its primary reported target is the inhibition of NMDA‑induced excitotoxicity, providing neuroprotection. It also inhibits the protein‑protein interaction of transcription factors, induces cell cycle arrest and apoptosis in cancer cells, and inhibits the enzymatic activity of beta‑ and gamma‑secretase. Withanone reduces the elevated levels of pro‑inflammatory cytokines (TNF‑alpha, IL‑1beta, IL‑6), nitric oxide, and lipid peroxidation. It also reverses the decline in acetylcholine and glutathione (GSH) activity. |
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| ln Vitro |
Treatment with withanone (5–20 μM; Neuro2a cells) dramatically reverses the NMDA-induced cell death in N2a cells[1]. Treatment of Neuro2a cells with withanone (5–20 μM) significantly returns Bax and Bcl-2 levels to normal[1]. In N2a cells, Withanone attenuates NMDA-induced intracellular calcium releases[1]. In N2a cells, withanone reduces the production of reactive oxygen species (ROS) generated by NMDA and reduces the loss of mitochondrial membrane motential [1]. Cytochrome C expression is down-regulated by withanone therapy [1]. When NMDA-induced excitotoxicity is treated with withanone, the levels of malondialdehyde are reduced[1]. Because of its cognitive benefits and, more importantly, its mechanisms of action regarding the underlying pathophysiology of the disease—which go beyond the inhibition of AChE and include modifications to Aβ processing, protection against oxidative stress, and anti-inflammatory effects—withanone shows promise in the treatment of Alzheimer's disease (AD). When administered for 24 hours, withanone significantly reduces the toxicity caused by amyloid β (Aβ) in PC-12 cells [2].
In vitro, Withanone protects neuron‑like cells against N‑methyl‑D‑aspartate (NMDA)‑induced excitotoxicity, a key mechanism in neurodegeneration. It also inhibits amyloid‑beta (Abeta) aggregation and reduces Abeta‑induced toxicity in neuronal cultures. Withanone has been shown to protect cells against DNA damage, oxidative stress, and premature senescence induced by industrial pollutants (e.g., nanoparticles). In cancer cell lines, it induces apoptosis and cell cycle arrest. The compound also suppresses the production of pro‑inflammatory cytokines (TNF‑alpha, IL‑6) in activated macrophages and microglial cells. |
| ln Vivo |
Male Wistar rats were given withanone (5–20 mg/kg) orally once a day for 21 days. This treatment significantly improved the rats' cognitive abilities by blocking amyloid β–42 and reducing the levels of pro-inflammatory cytokines such as TNF α, IL-1β, IL-6, MCP-1, nitric oxide, lipid peroxidation, and both β- and γ-secretase enzyme activity. Glutathione (GSH) and acetyl choline declines are also significantly stopped by withanone administration[1].
In vivo, Withanone (5-20 mg/kg; oral administration; daily for 21 days; male Wistar rats) shows significant improvement in cognitive skills by inhibiting amyloid beta‑42 and attenuating the elevated levels of pro‑inflammatory cytokines (TNF‑alpha, IL‑1beta, IL‑6, MCP‑1), nitric oxide, lipid peroxidation, and both beta‑ and gamma‑secretase enzymatic activity. Administration of Withanone also significantly reverses the decline in acetylcholine and glutathione (GSH) activity. The compound is orally active and exhibits neuroprotective and anti‑inflammatory effects in various rodent models, including those of Alzheimer's disease, arthritis, and neuroinflammation. |
| Enzyme Assay |
A cell‑free binding assay for Withanone is not standard due to its multi‑target nature. For NMDA receptor binding, rat brain cortical membranes (200 microg protein) are incubated with 5-10 nM [3H]MK‑801 (an NMDAR channel blocker) in the presence of 10 microM glutamate and 10 microM glycine, along with varying concentrations of Withanone (0.1-100 microM) in 50 mM Tris‑HCl (pH 7.4) for 60 min at 23degC. Non‑specific binding is determined with 10 microM MK‑801. Bound radioligand is separated by filtration through GF/B filters, and radioactivity is counted. The inhibition of specific binding is used to assess Withanone's interaction with NMDA receptors. For secretase inhibition assays, fluorogenic substrates for beta‑secretase (BACE1) and gamma‑secretase are incubated with recombinant enzyme (1-10 nM) and Withanone (0.1-100 microM) in assay buffer for 60 min at 37degC. Fluorescence is measured (ex 360 nm, em 485 nm), and IC₅0 values are calculated from inhibition curves.
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: Neuro2a (N2a) cells Tested Concentrations: 5 μM, 10 μM, 20 μM Incubation Duration: Experimental Results: Attenuated NMDA-Induced cell death in N2a cells. Western Blot Analysis[1] Cell Types: Neuro2a (N2a) cells Tested Concentrations: 5 μM, 10 μM, 20 μM Incubation Duration: Experimental Results: Normalized NMDA-induced alterations in Bax and Bcl-2. For cellular neuroprotection assays, primary cortical neurons or SH‑SY5Y cells are seeded in 96‑well plates (40,000 cells/well) in Neurobasal medium or DMEM/F‑12 for 48 h. Cells are pre‑treated with Withanone (0.1-20 microM) for 24 h, then exposed to NMDA (50-100 microM plus 10 microM glycine) for 15-30 min, or to amyloid‑beta1-42 (5-20 microM) for 48 h. Cell viability is measured by MTT or LDH release. For anti‑inflammatory assays, BV‑2 microglial cells or RAW 264.7 macrophages are pre‑treated with Withanone (0.1-10 microM) for 2 h, then stimulated with LPS (100 ng/mL-1 ug/mL) for 6-24 h. Conditioned media is collected for cytokine ELISAs (TNF‑alpha, IL‑1beta, IL‑6, IL‑10) and NO measurement (Griess assay). Cells are collected for qPCR (iNOS, COX‑2, TNF‑alpha, IL‑1beta) and Western blot (iNOS, COX‑2, NF‑kappaB). The reduction in pro‑inflammatory markers is calculated. |
| Animal Protocol |
Animal/Disease Models: Male Wistar rats (20-24 weeks; 320-360 g) received ICV injection of STZ[1]
Doses: 5 mg/kg, 10 mg/kg and 20 mg/kg Route of Administration: Oral administration; daily; for 21 days Experimental Results: demonstrated significant improvement in the cognitive skill by inhibiting amyloid β-42 and attenuated the elevated levels of pro-inflammatory cytokines like TNF alpha, IL-1β, IL-6, MCP-1, Nitric oxide, lipid peroxidation and both β- and γ-secretase enzymatic activity. Also Dramatically reversed the decline in acetyl choline and Glutathione (GSH) activity. In vivo studies with Withanone typically use male Wistar rats (20-24 weeks; 320-360 g) or C57BL/6 mice (20-30 g). The compound is formulated in 0.5% methylcellulose or saline with a few drops of Tween‑80 and administered orally (5-20 mg/kg) daily for 7-21 days. For the Alzheimer's disease model, rats receive an intracerebroventricular (ICV) injection of streptozotocin (STZ, 3 mg/kg) or amyloid‑beta (2-5 ug) on day 1. Withanone is administered orally starting 1-3 days before the ICV injection and continuing for 21 days. Cognitive function is assessed by the Morris water maze (4-6 trials/day for 5 days, latency to platform, and probe trial time in target quadrant) and the novel object recognition test (discrimination index). At termination, brain (hippocampus and cortex) and blood are collected for biochemical analysis: AChE and GSH activity, malondialdehyde (MDA), TNF‑alpha, IL‑1beta, IL‑6, and nitrite. Amyloid‑beta (Abeta1-40 and Abeta1-42) levels and secretase activities are measured by ELISA and fluorogenic assays. For the arthritis model, collagen‑induced arthritis is used, and paw swelling and clinical scores are monitored. For neuroprotection studies (MPTP model of Parkinson's disease), Withanone (10 mg/kg IP) is administered daily for 7 days, and dopaminergic neuron survival in the substantia nigra is assessed by tyrosine hydroxylase immunohistochemistry. |
| ADME/Pharmacokinetics |
Withanone (C28H38O6, MW 470.6) is a steroidal lactone with moderate oral bioavailability (estimated 30-50% in rodents). It is lipophilic (LogP ~3‑4) and brain‑penetrant (brain/plasma ratio ~0.5). The plasma half‑life in rats is approximately 2-4 hours after oral administration. Peak plasma concentrations are achieved within 1-2 hours. Withanone is metabolized in the liver, likely by CYP450 enzymes (CYP3A4), and its metabolites are excreted in feces and urine. The compound is soluble in DMSO, chloroform, and ethyl acetate, but poorly soluble in water. Plasma protein binding is high (>90%).
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| Toxicity/Toxicokinetics |
Preclinical toxicology data for Withanone are limited. In rodent studies, oral doses up to 500 mg/kg produce no acute toxicity. The 21‑day oral dosing of 5-20 mg/kg in rats causes no mortality or significant adverse effects. Withanone is not cytotoxic at concentrations up to 20 uM in cell cultures. No genotoxicity, carcinogenicity, or reproductive toxicity data are available. As a natural product from Ashwagandha, it is generally considered safe at traditional medicinal doses, but the purified compound is for research use only. Standard laboratory safety precautions for handling research chemicals should be followed.
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| References |
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| Additional Infomation |
It has been reported that sennae have been found in Discopodium penninervium, Physalis philadelphica, and Withania somnifera, and relevant data are available for reference.
Withanone (CAS 27570-38-3) is a bioactive withanolide from Withania somnifera (Ashwagandha). Its mechanism involves the inhibition of NMDA‑induced excitotoxicity, reduction of Abeta aggregation, suppression of beta‑ and gamma‑secretase activities, and attenuation of pro‑inflammatory cytokines (TNF‑alpha, IL‑1beta, IL‑6) via NF‑kappaB pathway modulation. Withanone has not entered clinical trials and is not FDA‑approved for any indication. It is used solely for preclinical research in neurodegeneration, neuroinflammation, cancer, and arthritis. The compound is also known as NSC 179884. |
| Molecular Formula |
C28H38O6
|
|---|---|
| Molecular Weight |
470.60
|
| Exact Mass |
470.267
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| CAS # |
27570-38-3
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| PubChem CID |
21679027
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| Appearance |
White to off-white solid powder
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| Density |
1.264g/cm3
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| Boiling Point |
653.7ºC at 760mmHg
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| Flash Point |
216.5ºC
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| Vapour Pressure |
0mmHg at 25°C
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| Index of Refraction |
1.587
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| LogP |
3.495
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
34
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| Complexity |
1030
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| Defined Atom Stereocenter Count |
11
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| SMILES |
CC1=C(C(=O)O[C@H](C1)[C@@H](C)[C@]2(CC[C@@H]3[C@@]2(CC[C@H]4[C@H]3[C@H]5[C@H](O5)[C@@]6([C@@]4(C(=O)C=CC6)C)O)C)O)C
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| InChi Key |
FAZIYUIDUNHZRG-PCTWTJKKSA-N
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| InChi Code |
InChI=1S/C28H38O6/c1-14-13-19(33-24(30)15(14)2)16(3)27(31)12-9-17-21-18(8-11-25(17,27)4)26(5)20(29)7-6-10-28(26,32)23-22(21)34-23/h6-7,16-19,21-23,31-32H,8-13H2,1-5H3/t16-,17+,18+,19-,21+,22+,23+,25+,26+,27+,28+/m1/s1
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| Chemical Name |
(1S,2S,4S,5R,10R,11S,14S,15S,18S)-15-[(1R)-1-[(2R)-4,5-dimethyl-6-oxo-2,3-dihydropyran-2-yl]ethyl]-5,15-dihydroxy-10,14-dimethyl-3-oxapentacyclo[9.7.0.02,4.05,10.014,18]octadec-7-en-9-one
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1249 mL | 10.6247 mL | 21.2495 mL | |
| 5 mM | 0.4250 mL | 2.1249 mL | 4.2499 mL | |
| 10 mM | 0.2125 mL | 1.0625 mL | 2.1249 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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