Withanolide B is a natural bioactive compound found in W. somnifera with anti-aging, neuroprotective, anti-arthritic, and anti-cancer effects. It promotes osteogenic differentiation of hBMSCs via regulating ERK1/2 and Wnt/β-catenin signaling pathways.
Physicochemical Properties
| Molecular Formula | C28H38O5 |
| Molecular Weight | 454.59832 |
| Exact Mass | 454.272 |
| CAS # | 56973-41-2 |
| PubChem CID | 14236711 |
| Appearance | White to off-white solid powder |
| Density | 1.206 |
| LogP | 4.38 |
| Hydrogen Bond Donor Count | 1 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 2 |
| Heavy Atom Count | 33 |
| Complexity | 981 |
| Defined Atom Stereocenter Count | 11 |
| SMILES | CC1=C(C(=O)O[C@H](C1)[C@@H](C)[C@H]2CC[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)C |
| InChi Key | ZTEVDTFJUUJBLP-MBMSZCMESA-N |
| InChi Code | InChI=1S/C28H38O5/c1-14-13-20(32-25(30)15(14)2)16(3)17-8-9-18-22-19(10-12-26(17,18)4)27(5)21(29)7-6-11-28(27,31)24-23(22)33-24/h6-7,16-20,22-24,31H,8-13H2,1-5H3/t16-,17+,18-,19-,20+,22-,23-,24-,26+,27-,28-/m0/s1 |
| Chemical Name | (1S,2S,4S,5R,10R,11S,14R,15R,18S)-15-[(1S)-1-[(2R)-4,5-dimethyl-6-oxo-2,3-dihydropyran-2-yl]ethyl]-5-hydroxy-10,14-dimethyl-3-oxapentacyclo[9.7.0.02,4.05,10.014,18]octadec-7-en-9-one |
| 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 |
| 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
| ln Vitro | In hBMSCs, withanolide B (1-100 nM; 3-5 days) dramatically boosts the expression of the COL1A1 and RUNX2 genes and proteins [1]. Alkaline phosphatase (ALP) activity and the production of extracellular matrix calcium deposits are both enhanced by withanolide B (1–100 nM; 11 or 3 days) [1]. Withanolide B (1-100 nM; 3-5 days) raises the levels of active β-catenin and p-ERK protein expression in hBMSCs [1]. In SK-N-SH cells, withanolide B (10-100 μM; 48 hours) reduces the toxicity caused by Aβ42 aggregation [2]. |
| ln Vivo | Withanolide B (10 mg/kg; local injection) can improve bone repair in a rat tibial defect model [1]. |
| Cell Assay |
Cell viability assay [1] Cell Types: human bone mesenchymal stem cells Tested Concentrations: 1, 10, 100 nM Incubation Duration: 3, 5 days Experimental Results: The expression of COL1A1 and RUNX2 increased Dramatically. |
| Animal Protocol |
Animal/Disease Models: Male SD (SD (Sprague-Dawley)) rats with tibial defects (8 weeks, 200 g) [1] Doses: 10 mg/kg Route of Administration: At various time points (0, 3, 5, 7, and 9 days) Experimental Results: Increased trabecular number, trabecular thickness, and cortical bone thickness. |
| References |
[1]. Withanolide B promotes osteogenic differentiation of human bone marrow mesenchymal stem cells via ERK1/2 and Wnt/β-catenin signaling pathways. Int Immunopharmacol. 2020 Nov;88:106960. [2]. Improving the inhibition of β-amyloid aggregation by withanolide and withanoside derivatives. Int J Biol Macromol. 2021 Mar 15;173:56-65. [3]. Enhanced biosynthesis of withanolides by elicitation and precursor feeding in cell suspension culture of Withania somnifera (L.) Dunal in shake-flask culture and bioreactor. PLoS One. 2014 Aug 4;9(8):e104005. |
| Additional Infomation |
Withanolide B is a withanolide. Withanolide B has been reported in Withania coagulans, Datura metel, and Withania somnifera with data available. |
Solubility Data
| 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 |
| 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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1997 mL | 10.9987 mL | 21.9974 mL | |
| 5 mM | 0.4399 mL | 2.1997 mL | 4.3995 mL | |
| 10 mM | 0.2200 mL | 1.0999 mL | 2.1997 mL |