β-Sitosterol (also known as SKF 14463), a lipid regulating agent, is one of several natural phytosterols (plant sterols) with chemical structures similar to that of cholesterol. It is a typical plant steroid with anticholesteremic properties. Recent research has demonstrated that β-Sitosterol causes G2/M arrest, endoreduplication, and apoptosis via the Bcl-2 and PI3K/Akt signaling pathways.

Physicochemical Properties

Molecular Formula	C29H50O
Molecular Weight	414.71
Exact Mass	414.386
Elemental Analysis	C, 83.99; H, 12.15; O, 3.86
CAS #	83-46-5
Related CAS #	83-46-5
PubChem CID	222284
Appearance	White to off-white solid powder
Density	1.0±0.1 g/cm3
Boiling Point	501.9±19.0 °C at 760 mmHg
Melting Point	139-142 ºC
Flash Point	220.4±13.7 °C
Vapour Pressure	0.0±2.9 mmHg at 25°C
Index of Refraction	1.521
LogP	10.73
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	1
Rotatable Bond Count	6
Heavy Atom Count	30
Complexity	634
Defined Atom Stereocenter Count	9
SMILES	C[C@@]12[C@@H]([C@H](C)CC[C@@H](CC)C(C)C)CC[C@H]1[C@@H]1CC=C3C[C@H](CC[C@]3(C)[C@H]1CC2)O
InChi Key	KZJWDPNRJALLNS-VJSFXXLFSA-N
InChi Code	InChI=1S/C29H50O/c1-7-21(19(2)3)9-8-20(4)25-12-13-26-24-11-10-22-18-23(30)14-16-28(22,5)27(24)15-17-29(25,26)6/h10,19-21,23-27,30H,7-9,11-18H2,1-6H3/t20-,21-,23+,24+,25-,26+,27+,28+,29-/m1/s1
Chemical Name	(3S,8S,9S,10R,13R,14S,17R)-17-[(2R,5R)-5-ethyl-6-methylheptan-2-yl]-10,13-dimethyl-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-ol
Synonyms	(-)-beta-Sitosterol; 22,23-Dihydrostigmasterol; 24-alpha-Ethylcholesterol; AI3-26020; alpha-Dihydrofucosterol; Rhamnol; Angelicin; beta-Sitosterol; CCRIS 5529; Azuprostat; Cinchol; Cupreol; Harzol; Nimbosterol; Prostasal; Quebrachol; Triastonal
HS Tariff Code	2934.99.03.00
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)

Biological Activity

Targets	lipid regulating agent
ln Vitro	Beta-Sitosterol is one of the most abundant dietary phytosterols. According to one study, beta-sitosterol has the ability to prevent leukemia, ovarian cancer, breast, prostate, colon, lung, stomach, and colon cancer.
ln Vivo	The total cells and eosinophils in the bronchoalveolar lavage (BAL) fluid markedly decreased (p<0.05) after L-BS or beta-sitosterol (BS) administration (1 mg/kg; i.p.), and the ROS production also decreased in comparison to the asthma control. Histopathological features were detected by performing histochemistry, including H&E and alcian blue & P.A.S staining. Both L-BS and beta-sitosterol (BS) mitigated the inflammation by eosinophil infiltration and mucus hypersecretion by goblet hyperplasia. These effects of L-BS were superior to those of BS. L-BS and BS inhibited the increased mRNA and protein expression of IL-4 and IL-5 in the lung tissue and BAL fluid, respectively. The IgE concentration in the BAL fluid and serum was measured by performing ELISA and the ovalbumin-specific IgE in the BAL fluid was uniquely inhibited by L-BS (p<0.05). The splenocytes were isolated from the normal and asthmatic mice and incubated in the absence and presence of 100 microg/ml ovalbumin, respectively. L-BS blocked the survival rate of the splenocytes of the mice (p<0.01). This finding indicates the possibility of L-BS and BS as potential therapeutic molecules in asthma and may contribute to the need to improve current therapeutic drugs. [2]
Cell Assay	Viability of mouse splenocytes [2] We performed MTT assay and apoptosis assay to determine the cell viability with using the MTT assay kit and annexinV-fluorescein isothiocyanate (FITC) apoptosis detection kit. The splenocytes were isolated from the spleens of normal and asthmatic control mice by syringe pumping. After three washings with 10 ml of DMEM supplemented with antibiotics–antimycotics, the splenocytes were incubated with 3 ml of RBC lysis buffer for 10 min at room temperature, and then this was washed twice with 10 ml of the wash medium. 2 × 105 splenocytes in 100 μl of DMEM medium containing 10% FBS were seeded onto a 96-well culture plate. L-BS, BS or dexamethasone (1 μg/ml) in the absence or presence of 100 μg/ml ovalbumin was added to the individual well and then the plate was incubated for 48 h at 37 °C in a CO2 incubator. After the addition of 10 μl of MTT solution in each well, the plate was incubated at 37 °C for 4 h in a CO2 incubator and 100 μl of solubilization solution was added to each well for MTT assay. After 24 h incubation, the absorbance was measured by using an ELISA reader at 550 nm. For the apoptosis assay, the cells were harvested and resuspended in binding buffer. Annexin V-FITC and PI were added and incubated for 15 min at room temperature. The cells were analyzed by FACSort cytofluorimeter using CellQuest software. Negative cells against annexin V and PI staining were considered as viable or non-apoptotic cells.[2]
Animal Protocol	Induction of asthma in mice[2] Six to eight-week-old female BALB/c mice were obtained from Daehan Biolink Co. LTD. They were maintained in an air-conditioned room. The room temperature (about 22 ± 1 °C) and humidity (about 55 ± 10%) were automatically controlled. The mice were divided into five groups (n = 5), and airway inflammation was induced in four groups. Each mouse was immunized through intraperitoneal (i.p.) injection with 20 μg of chicken OVA and 1 mg of aluminum hydroxide on days 1 and 14, as shown in Fig. 2. The mice were exposed to a 5% ovalbumin solution aerosolized using an ultrasonic nebulizer for 1 h per day from days 21 to 27 after the second sensitization. The mice were placed in a Plexiglass chamber (30 × 30 × 15 cm3) that contained small ventilation holes on one side during the inhalation challenge. The aerosol was generated with a nebulization rate of 1 ml/min. Three groups of asthma-induced mice were treated through i.p. injection with 1 mg/kg of L-BS, BS or dexamethasone between days 14 and 27. Both L-BS and BS dissolved in DMSO diluted less than 1/100 by phosphate-buffered saline (PBS). The negative control group was sensitized and challenged with PBS without drug administration.
Toxicity/Toxicokinetics	mouse LD oral >25 gm/kg Cancer Letters, 127(135), 1998 [PMID:9619869]
References	[1]. Beta-Sitosterol: A Promising but Orphan Nutraceutical to Fight Against Cancer. Nutr Cancer. 2015;67(8):1214-20. [2]. Int Immunopharmacol. 2007 Dec 5;7(12):1517-27. doi: 10.1016/j.intimp.2007.07.026.
Additional Infomation	Sitosterol is a member of the class of phytosterols that is stigmast-5-ene substituted by a beta-hydroxy group at position 3. It has a role as a sterol methyltransferase inhibitor, an anticholesteremic drug, an antioxidant, a plant metabolite and a mouse metabolite. It is a 3beta-sterol, a stigmastane sterol, a 3beta-hydroxy-Delta(5)-steroid, a C29-steroid and a member of phytosterols. It derives from a hydride of a stigmastane. Active fraction of Solanum trilobatum; reduces side-effects of radiation-induced toxicity. Beta-Sitosterol has been reported in Sambucus chinensis, Erythrophleum fordii, and other organisms with data available. All the currently available cancer therapeutic options are expensive but none of them are safe. However, traditional plant-derived medicines or compounds are relatively safe. One widely known such compound is beta-sitosterol (BS), a plant derived nutrient with anticancer properties against breast cancer, prostate cancer, colon cancer, lung cancer, stomach cancer, ovarian cancer, and leukemia. Studies have shown that BS interfere with multiple cell signaling pathways, including cell cycle, apoptosis, proliferation, survival, invasion, angiogenesis, metastasis and inflammation. Most of the studies are incomplete partly due to the fact that BS is relatively less potent. But the fact that it is generally considered as nontoxic, the opposite of all currently available cancer chemo-therapeutics, is missed by almost all research communities. To offset the lower efficacy of BS, designing BS delivery for "cancer cell specific" therapy hold huge potential. Delivery of BS through liposome is one of such demonstrations that has shown to be highly promising. But further research did not progress neither in the field of drug delivery of BS nor in the field on how BS mediated anticancer activities could be improved, thus making BS an orphan nutraceutical. Therefore, extensive research with BS as potent anticancer nutraceutical is highly recommended. [1] Asthma is a disease marked by chronic lung inflammation and the number of patients suffering from asthma increases annually. Both beta-sitosterol (BS) and beta-sitosterol glucoside exist in a variety of plants and have anti-tumor, anti-microbial, and immunomodulatory activities. However, the precise role of BS and beta-sitosterol glucoside in asthma has not been well understood. The aim of this study was to investigate the inhibitory effects of BS and lactose-BS (L-BS) on the pathophysiological process in ovalbumin-induced asthmatic mice. The total cells and eosinophils in the bronchoalveolar lavage (BAL) fluid markedly decreased (p<0.05) after L-BS or BS administration (1 mg/kg; i.p.), and the ROS production also decreased in comparison to the asthma control. Histopathological features were detected by performing histochemistry, including H&E and alcian blue & P.A.S staining. Both L-BS and BS mitigated the inflammation by eosinophil infiltration and mucus hypersecretion by goblet hyperplasia. These effects of L-BS were superior to those of BS. L-BS and BS inhibited the increased mRNA and protein expression of IL-4 and IL-5 in the lung tissue and BAL fluid, respectively. The IgE concentration in the BAL fluid and serum was measured by performing ELISA and the ovalbumin-specific IgE in the BAL fluid was uniquely inhibited by L-BS (p<0.05). The splenocytes were isolated from the normal and asthmatic mice and incubated in the absence and presence of 100 microg/ml ovalbumin, respectively. L-BS blocked the survival rate of the splenocytes of the mice (p<0.01). This finding indicates the possibility of L-BS and BS as potential therapeutic molecules in asthma and may contribute to the need to improve current therapeutic drugs. [2]

Solubility Data

Solubility (In Vitro)

DMSO:<1 mg/mL Water: <1 mg/mL Ethanol: ~5 mg/mL (12.1 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 20 mg/mL (48.23 mM) in 0.5% CMC-Na/saline water (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 1 mg/mL (2.4 mM) (saturation unknown) in 10% EtOH + + 40% PEG300 + 5% Tween80 + + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution. For example, if 1 mL of working solution is to be prepared, you can take 100 μL of 25 mg/mL EtOH + stock solution and add to 400 μL of PEG300, mix well; Then add 50 μL of Tween 80 to the above solution, mix well; Finally, add 450 μL of saline to the above solution, mix well. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Solubility in Formulation 3: ≥ 1 mg/mL (2.4 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. For example, if 1 mL of working solution is to be prepared, you can take 100 μL of 25 mg/mL EtOH + stock solution and add to 900 μL of 20% SBE-β-CD in saline, mix well. 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 4: ≥ 1 mg/mL (2.4 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 take 100 μL of 25 mg/mL EtOH + stock solution and add to 900 μL of corn oil, mix well (clear solution). Solubility in Formulation 5: ~5 mg/mL (12.1 mM) in 15% Cremophor EL + + 85% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution. Solubility in Formulation 6: ~10 mg/mL (24.1 mM) in Corn Oil , clear solution.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.4113 mL	12.0566 mL	24.1132 mL
	5 mM	0.4823 mL	2.4113 mL	4.8226 mL
	10 mM	0.2411 mL	1.2057 mL	2.4113 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.