| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| Other Sizes |
Purity: ≥98%
| Targets |
NF-κB; vimentin
Withaferin A targets multiple molecular targets. It prevents NF-κB activation by inhibiting IKKβ activation via a thioalkylation-sensitive redox mechanism. It also targets the intermediate filament protein vimentin, and inhibits endothelial protein C receptor (EPCR) shedding. Additionally, it inhibits chymotrypsin-like activity of the 20S proteasome. It is a cell-permeable and potent angiogenesis inhibitor. |
|---|---|
| ln Vitro |
Withaferin A inhibits NF-kB activation effectively and has anti-inflammatory properties by preventing Ik-B kinase beta from being activated by TNF through a thioalkylation-sensitive redox mechanism[1].
Withaferin A has anticancer properties as well. The IF protein vimentin is the target of withaferin A, which also causes vimentin filaments to aggregate in bovine aortic endothelial cells (BAECs) at 3 μM and fragment in endothelial cells at 10 μM[2]. The tumorigenic potential of ALDH1 positive cancer stem cells (CSCs) is decreased in a dose-dependent manner by withaferin A (0.5, 1.5 μM) alone or in combination with cisplatin (CIS)[3]. In vitro, withaferin A inhibits endothelial cells (HUVEC) sprouting with an IC50 of 12 nM. It inhibits proliferation of human endometrial cancer cells via transforming growth factor-β (TGF-β) signaling. It potently inhibits NF-κB activation by preventing TNF-induced activation of IKKβ. It inhibits α-actinin-1 and α4-crystallin. These in vitro activities demonstrate its potent antiangiogenic, anticancer, and anti-inflammatory effects. |
| ln Vivo |
In mice, withaferin A (2 mg/kg, i.p.) exhibits strong angiogenesis inhibitory activity through vimentin[2]. In tumors taken from mice, the combination of withaferin A (2 mg/kg) and cisplatin (CIS) controls the expression of the ALDH1 marker and inhibits the expression of securin[3].
In vivo, withaferin A has demonstrated antitumor and antiangiogenic activity. It inhibits endothelial cell sprouting in vivo. It has anti-inflammatory activity and has been shown to inhibit tumor growth in various animal models. However, the pharmacological and toxicological properties of withaferin A have not been fully investigated. Its in vivo efficacy supports its potential as a therapeutic agent for cancer and inflammatory diseases. |
| Enzyme Assay |
The in vitro enzyme/receptor binding assays for withaferin A include measurement of its ability to inhibit NF-κB activation. This is typically assessed by measuring its ability to prevent IKKβ activation in cell-free systems. Its inhibition of the chymotrypsin-like activity of the 20S proteasome can also be measured in cell-free assays. These assays provide a direct measure of the compound's activity against its targets.
|
| Cell Assay |
The RPMI1640 medium is used to support the ovarian epithelial cancer cell line A2780. It also contains insulin (5 g/mL), penicillin/streptomycin (100 IU/mL and 100 g/mL, respectively), and 10% fetal bovine serum (FBS) from Hyclone. In DMSO, other reagents such as withaferin A and cisplatin (CIS) are prepared. Each time it is made, cisplatin is fresh[3].
In vitro cellular assays for withaferin A assess its effects on endothelial cells, cancer cells, and immune cells. HUVEC sprouting assays are used to assess its antiangiogenic activity (IC50 = 12 nM). Its antiproliferative effects are assessed in cancer cell lines. NF-κB activation is measured in cells stimulated with TNF-α. These assays demonstrate the compound's functional activity in a cellular context. |
| Animal Protocol |
Mice: In the assay, mice with either a vimentin homozygous defect (Vim−/−) or a vimentin heterozygous defect (Vim−/−) on the 129/Svev background are used. The intraperitoneal (i.p.) injection of ketamine and xylazine is used to anesthetize mice between the ages of 4 and 6 weeks. Proparacain eye drop is used to topically anesthetize the corneas before 1 μL of diluted sodium hydroxide solution (0.15 M) is applied for 1 minute. The cornea is immediately thoroughly washed in saline solution, followed by the delicate removal of the corneal and limbal epithelium by scraping. Atropine eye drops are used topically to treat the cornea, and antibiotic eye ointments containing tobramycin and erythromycin are used to protect it. In the drug or control groups of mice, withaferin A or 12-D WS (2 mg/kg solubilized in DMSO) or vehicle (DMSO) are injected intraperitoneally (i.p.) after the mice have recovered from corneal injury, and then once a day for the following ten days. Eyes are removed from mice that have been killed humanely. In order to prepare the corneal buttons, the anterior segment of the eyes is divided in half. Corneal tissues are fixed in 100% acetone for 20 minutes, cleaned in PBS for 1 hour, and blocked for 18 hours at 4°C in 1% BSA-PBS. Cornea whole-mount staining involves soaking tissues in FITC-conjugated rat anti-mouse CD31 antibody (1:333 dilution in 1% BSA-PBS) for 12 hours, rinsing them off for 24 hours at 4°C in the same solution, and attaching them to glass slides with coverslips. By importing digital images into NIH ImageJ, fluorescent staining can be quantified after being visualized on a microscope[2].
In vivo animal studies for withaferin A have been conducted in various tumor models to assess its antitumor and antiangiogenic activity. It has been shown to inhibit endothelial cell sprouting in vivo. However, specific details of in vivo studies, such as the animal models used and the dosing regimens, are not extensively detailed in the available literature. Its pharmacological and toxicological properties have not been fully investigated. |
| ADME/Pharmacokinetics |
Specific pharmacokinetic data for withaferin A are not extensively detailed in the available literature. As a natural product, its oral bioavailability may be limited. However, its in vivo efficacy suggests that it reaches sufficient systemic concentrations to exert its effects. The compound's pharmacokinetic properties would be important for its development as a therapeutic agent, but specific data are not provided.
|
| Toxicity/Toxicokinetics |
Specific toxicity data for withaferin A are not extensively detailed in the available literature. The compound's pharmacological and toxicological properties have not been fully investigated. Caution should be used when handling this compound. As a potent NF-κB inhibitor and antiangiogenic agent, its toxicity profile would need to be carefully evaluated for therapeutic use. Comprehensive toxicological studies would be required for clinical development.
|
| References |
|
| Additional Infomation |
Withaferin A is a Withaferin with the structure 5,6:22,26-diepoxyergoster-2,24-diene-1,26-dione, substituted with hydroxyl groups at positions 4 and 27 (4β,5β,6β,22R stereoisomers). It was isolated from Physalis longifolia and exhibits cytotoxic activity. It can be used as an antitumor drug and an apoptosis inducer. Withaferin A is a δ-lactone, 4-hydroxysteroid, enone, ergosterane compound, secondary alcohol, Withaferin, 27-hydroxysteroid, primary alcohol, and epoxysteroid compound. It has been reported to exist in Iochroma gesnerioides, Withania coagulans, and several other organisms with relevant data.
Withaferin A is a steroidal lactone isolated from Ashwagandha (Withania somnifera) with potent anti-inflammatory, antitumor, and antiangiogenic activities. It inhibits NF-κB activation, targets vimentin, and inhibits EPCR shedding. It is a cell-permeable angiogenesis inhibitor with an IC50 of 12 nM for HUVEC sprouting. It is a research compound used for studying inflammation, cancer, and angiogenesis. It is not approved for clinical use and is intended for research purposes only. |
| Molecular Formula |
C28H38O6
|
|---|---|
| Molecular Weight |
470.5977
|
| Exact Mass |
470.266
|
| Elemental Analysis |
C, 71.46; H, 8.14; O, 20.40
|
| CAS # |
5119-48-2
|
| Related CAS # |
5119-48-2
|
| PubChem CID |
265237
|
| Appearance |
White to off-white solid powder
|
| Density |
1.3±0.1 g/cm3
|
| Boiling Point |
680.7±55.0 °C at 760 mmHg
|
| Melting Point |
252-253ºC
|
| Flash Point |
226.7±25.0 °C
|
| Vapour Pressure |
0.0±4.8 mmHg at 25°C
|
| Index of Refraction |
1.599
|
| LogP |
3.8
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
34
|
| Complexity |
999
|
| Defined Atom Stereocenter Count |
11
|
| SMILES |
O1[C@]2([H])C([H])([H])[C@@]3([H])[C@]4([H])C([H])([H])C([H])([H])[C@]([H])([C@]([H])(C([H])([H])[H])[C@@]5([H])C([H])([H])C(C([H])([H])[H])=C(C([H])([H])O[H])C(=O)O5)[C@@]4(C([H])([H])[H])C([H])([H])C([H])([H])[C@]3([H])[C@]3(C(C([H])=C([H])[C@@]([H])([C@]132)O[H])=O)C([H])([H])[H]
|
| InChi Key |
DBRXOUCRJQVYJQ-CKNDUULBSA-N
|
| InChi Code |
InChI=1S/C28H38O6/c1-14-11-21(33-25(32)17(14)13-29)15(2)18-5-6-19-16-12-24-28(34-24)23(31)8-7-22(30)27(28,4)20(16)9-10-26(18,19)3/h7-8,15-16,18-21,23-24,29,31H,5-6,9-13H2,1-4H3/t15-,16-,18+,19-,20-,21+,23-,24+,26+,27-,28+/m0/s1
|
| Chemical Name |
(1S,2R,6S,7R,9R,11S,12S,15R,16S)-6-hydroxy-15-[(1S)-1-[(2R)-5-(hydroxymethyl)-4-methyl-6-oxo-2,3-dihydropyran-2-yl]ethyl]-2,16-dimethyl-8-oxapentacyclo[9.7.0.02,7.07,9.012,16]octadec-4-en-3-one
|
| Synonyms |
NSC 101088; NSC101088; NSC-101088; NSC-273757; NSC273757; NSC 273757; Withaferin A
|
| 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 requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
DMSO: 50~94 mg/mL (106.3~199.75 mM)
Ethanol: ~4 mg/mL (~8.50 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 0.83 mg/mL (1.76 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 8.3 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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: ≥ 0.83 mg/mL (1.76 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 8.3 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. View More
Solubility in Formulation 3: ≥ 0.83 mg/mL (1.76 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| 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.
| NCT Number | Recruitment | interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
| NCT05610735 | Not yet recruiting | Drug: DOXIL Drug: Withaferin A |
Recurrent Ovarian Cancer | Sham Sunder Kakar | August 1, 2023 | Phase 1 Phase 2 |
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA