Product name: Stigmasterol glucoside
Synonym name:
Catalogue No.: BP3373
Cas No.: 19716-26-8
Formula: C₃₅H₅₈O₆
Mol Weight: 574.843
Botanical Source:
Physical Description: Powder
Type of Compound: Steroids

Purity: 95%~99%
Analysis Method: HPLC-DAD or/and HPLC-ELSD
Identification Method: Mass, NMR
Packing: Brown vial or HDPE plastic bottle

Storage: Store in a well closed container, protected from air and light. Put into refrigerate or freeze for long term storage.
Whenever possible, you should prepare and use solutions on the same day. However, if you need to make up stock solutions in advance, we recommend that you store the solution as aliquots in tightly sealed vials at -20℃. Generally, these will be useable for up to two weeks.

The product could be supplied from milligrams to grams, up to kilograms
Inquire for bulk scale.

Descriptions:
Stigmasterol glucoside, a constituent of Adenanthera pavonina seed and leaf, may be a plant anticancer agent.[1,2]

References:
[1] Nigam S K, Misra G, Mitra C R. Planta Medica, 1973, 23(2):145-8.
[2] Arisawa M, Kinghorn A D, Cordell G A, et al. Planta Medica, 1985, 51(6):544-5.
[3] Chen Z, Bi X, Luo W, et al. J Guangxi Medical University, 2014, 17(1):74-6.

HNMR of Stigmasterol glucoside

HPLC of Stigmasterol glucoside

Storage conditions：Short-term storage at 2~8℃, long-term storage at -20 ~ -80℃

Introduction/Overview

Cardiovascular disease (CVD) is the leading cause of death and disability worldwide, with dyslipidemia, particularly hypercholesterolemia, being its core risk factor. Although statins have achieved great success as first-line lipid-lowering drugs, their muscle toxicity, risk of liver injury, and the phenomenon of "statin intolerance" in some patients have prompted researchers to continuously search for safer, multi-target alternative or adjuvant treatment strategies from natural products. Plant sterols and their glycoside derivatives, as secondary metabolites widely present in the plant kingdom, have long been of great concern due to their structural similarity with cholesterol and clear cholesterol lowering activity. Stigmasterol glucoside (SG), as an important glycosylation form of stigmasterol, not only retains the physiological activity of parent sterols, but also changes its physicochemical properties and bioavailability due to the introduction of glycosylation, demonstrating unique pharmacological potential. In recent years, studies have revealed that SG is not only a classic 5 α - reductase inhibitor, but also shows significant activity in regulating key targets of lipid metabolism, laying a scientific foundation for its application in the field of reducing blood lipids and anti atherosclerosis. This article aims to systematically review the chemical properties, plant sources, pharmacological activities of SG, especially its multi-target lipid-lowering mechanism, drug evaluation, and clinical application prospects, in order to provide comprehensive academic references for the deep development of this natural product.

Chemical structure and physicochemical properties

Dougesterol glucoside (CAS: 19716-26-8) is a steroid saponin compound with a molecular formula of C35H58O6 and a molecular weight of 574.8430. Its chemical structure consists of two parts: a hydrophobic sterol core and a hydrophilic glycoside group.

1. Sterol nucleus Stigmasterol, belonging to Δ 5-steronols. Its structural feature is the presence of a double bond at the C-5 position and an ethyl substitution at the C-24 position (24 ethyl - Δ 5,22-cholesterol-3 β - ol), which distinguishes it from animal derived cholesterol (hydrogen at the C-24 position) and sitosterol (ethyl at the C-24 position, but without a double bond at C-22), and is also the structural basis for its specific biological activity.
2. Glycoside part A β - D-glucopyranosyl group is connected to the C-3 hydroxyl group of the sterol parent nucleus through a glycosidic bond. The introduction of sugar groups significantly enhances the polarity of the molecule.

Based on the above structure, SG exhibits typical amphiphilic molecular characteristics, which directly affect its physicochemical properties:
* Lipid water partition coefficient (LogP)The calculated value is about 5.63, indicating that the compound has high lipophilicity, mainly due to its large steroid nucleus structure. However, compared to its aglycone sitosterol (with a higher LogP), the introduction of glucose groups reduces its LogP value and increases its dispersibility in the aqueous phase.
* Topological Polarity Surface Area (TPSA)Approximately 99.38 Å ², mainly contributed by multiple hydroxyl oxygen atoms on the glucose unit. A higher TPSA value indicates that the molecule has a certain polarity.
* Water solubility Extremely low, with an experimental or predicted value of about 0.003 mg/mL, it belongs to insoluble compounds. This limits its direct application in aqueous media and is a key issue that needs to be addressed in formulation development.
* spectral characteristics In the nuclear magnetic resonance hydrogen spectrum, characteristic proton signals of steroid nuclei (such as C-18, C-19 angle methyl, C-6 ene hydrogen), as well as glucose end group protons (δ~4.2-4.4 ppm, d, J=7-8 Hz, indicating β configuration) and other glycosyl proton signals can be observed. Excimer ion peaks such as [M+H]+or [M+Na]+can be observed in mass spectrometry, as well as glycoside fragment peaks produced by the loss of glucose groups (-162 Da).

Plant sources and extraction methods

SG is widely distributed in various plants such as leguminous, Solanaceae, Lamiaceae, and Scrophulariaceae, and often coexists with other sterol glycosides such as β - sitosterol glucoside.

• Main plant sources：

  • Leguminous plants As the source of the name "stigmasterol", soybean（Glycine max）Its products are one of the most abundant natural sources of SG. In addition, chickpea, pea and astragalus are also high in content.
  • medicinal plants Many traditional medicinal plants contain SG and may be related to their medicinal properties. For example, Rehmannia glutinosa（Rehmannia glutinosa）, Radix Rehmanniae（Digitalis purpurea However, attention should be paid to the toxicity of cardiac glycosides and centella asiatica（Centella asiatica）Heart piercing lotus（Andrographis paniculata）Wait.
  • Other edible plants Such as pumpkin seeds, avocados, etc.

• Extraction and Separation Methods：

  1. Extract Organic solvent reflux extraction or ultrasound assisted extraction are commonly used. Due to the low polarity of SG, medium polarity solvents such as methanol, ethanol, ethyl acetate, or their mixed solvents with water are commonly used. In recent years, green extraction technologies such as supercritical CO2 extraction (which requires the addition of entrainers such as ethanol) have also been applied.
  2. Enrichment and Purification After defatting with petroleum ether, the crude extract can be preliminarily separated using the amphiphilic properties of SG. Common methods include:
     • Silica gel column chromatography Using chloroform methanol gradient elution, SG usually flows out in a higher proportion of methanol (such as chloroform: methanol=8:2 to 7:3).
     • Reverse phase chromatography C18 bonded phase column with methanol water or acetonitrile water as mobile phase is an effective method for obtaining high-purity SG.
     • Liquid liquid distribution Enrich by utilizing its distribution difference in n-butanol and water.
  3. appraisal Combined with thin-layer chromatography (TLC, commonly using 10% ethanol sulfate for color development), high-performance liquid chromatography (HPLC, commonly using evaporative light scattering detector ELSD or mass spectrometry detector), as well as nuclear magnetic resonance (NMR) and mass spectrometry (MS) for structural confirmation.

Pharmacological activity research

The pharmacological activity research of SG has expanded from early single target to multi-faceted biological effect evaluation, and its core activities can be summarized as follows:

1. Hypolipidemic and antiatherosclerotic activity This is currently the most concerned pharmacological effect of SG. In vitro and in vivo studies have shown that SG can effectively reduce the levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and triglycerides (TG) in the serum of high-fat diet induced ApoE -/- mice, while increasing high-density lipoprotein cholesterol (HDL-C). Its strength of action may be weaker than statins, but its multi-target properties show advantages. In addition, SG can reduce the area of atherosclerotic plaque, improve vascular endothelial function, inhibit vascular inflammation, and play a comprehensive anti atherosclerosis effect.

2. 5 α - reductase inhibitory activity SG is a natural 5 α - reductase inhibitor with an IC50 value of 27.2 µ M. 5 α - reductase can convert testosterone into the more active dihydrotestosterone (DHT), which is closely related to benign prostatic hyperplasia (BPH) and androgenic alopecia. SG can potentially be used to treat BPH and hair loss by inhibiting the enzyme and reducing DHT production.

3. Anti inflammatory and antioxidant activity：

   • anti-inflammatory SG can significantly inhibit the excessive production of nitric oxide (NO), prostaglandin E2 (PGE2), and pro-inflammatory cytokines (such as TNF - α, IL-1 β, IL-6) induced by lipopolysaccharide (LPS) and other factors in macrophages. The mechanism involves inhibiting the activation of nuclear factor kappa B (NF - κ B) and mitogen activated protein kinases (MAPKs) signaling pathways.
   • antioxidant SG can eliminate free radicals such as DPPH and ABTS, and enhance the intracellular antioxidant defense system, such as upregulating the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH Px), and reducing the level of malondialdehyde (MDA). Its antioxidant effect helps alleviate cellular damage and inflammatory response induced by oxidative stress.

4. Antitumor activity: Preliminary studies have shown that SG can inhibit the proliferation of many cancer cell lines (such as breast cancer MCF-7, liver cancer HepG2, colon cancer HT-29, etc.), and can induce apoptosis and cell cycle arrest. Its anti-tumor mechanism may be related to regulating the Bcl-2/Bax ratio, activating the caspase cascade, and inhibiting the PI3K/Akt/mTOR signaling pathway. However, its anti-tumor activity is mostly in the stage of cell experiments, and its in vivo potency and selectivity need further evaluation.

5. Other activities There are also reports that SG has potential activities such as neuroprotection, liver protection, and immune regulation, but research is not yet sufficient.

Mechanism of action and molecular targets

The lipid-lowering effect of SG is not achieved through a single pathway, but through the coordinated regulation of lipid metabolism homeostasis through multiple targets and links. Its core molecular targets and mechanisms are as follows:

1. Inhibit cholesterol synthesis SG can inhibit the activity of hydroxymethylglutaryl-CoA reductase (HMGCR). HMGCR is the rate limiting enzyme for cholesterol synthesis in the body and a classic target for statins. SG may reduce endogenous cholesterol synthesis by modulating or affecting its expression through allosteric regulation, thereby feedback upregulating the expression of low-density lipoprotein receptor (LDLR) in liver cells.
2. Upregulation of low-density lipoprotein receptor (LDLR)LDLR is a key membrane receptor for clearing circulating LDL-C. SG promotes the transcription and expression of LDLR genes by activating peroxisome proliferator activated receptor alpha (PPARA) or indirectly by reducing intracellular cholesterol levels, accelerating liver uptake and clearance of LDL-C.
3. Regulating apolipoprotein and lipoprotein metabolism：
   • Inhibition of Proprotein Converting Enzyme Subtilisin 9 (PCSK9)PCSK9 can bind and promote lysosomal degradation of LDLR. Research has shown that SG may downregulate the expression of PCSK9, thereby stabilizing LDLR protein levels and enhancing their function.
   • Affects apolipoprotein B (APOB) and apolipoprotein E (APOE)APOB is the main structural protein of LDL, and APOE plays a crucial role in the clearance of chylomicrons and VLDL residues. SG may regulate triglyceride rich lipoprotein metabolism by affecting their expression or post-translational modifications.
4. Activate PPARA pathway PPARA is a key nuclear receptor that regulates fatty acid oxidation, lipoprotein metabolism, and inflammation. SG, as a potential PPARA agonist, can promote the expression of fatty acid β - oxidation related genes (such as CPT1A) and reduce TG levels after activation; Simultaneously inducing the expression of lipoprotein lipase (LPL) and promoting the breakdown of TG rich lipoproteins; It can also have anti-inflammatory effects.
5. Inhibition of cholesterol ester transfer protein (CETP)CETP promotes the transfer of cholesterol esters from HDL to LDL/VLDL, reducing HDL-C levels. SG may inhibit the activity of CETP, thus maintaining or increasing the level of HDL-C with cardiovascular protection, and reducing LDL particles that cause atherosclerosis.
6. Anti inflammatory and antioxidant mechanisms The realization of its lipid-lowering benefits cannot be achieved without the assistance of anti-inflammatory and antioxidant effects. Reduce vascular wall inflammation by inhibiting pathways such as NF - κ B and MAPK; By scavenging free radicals and enhancing antioxidant enzymes, LDL is protected from oxidative modification (ox LDL), which is a key factor in the initiation of atherosclerosis.

To sum up, SG cooperatively plays the role of lowering blood lipid and anti atherosclerotic hardening through multiple network mechanisms of "inhibiting synthesis (HMGCR), promoting clearance (LDLR ↑, PCSK9 ↓), improving transport (CETP ↓, APOB/E), promoting oxidation (PPARA ↑)" and "anti-inflammatory and antioxidant".

Evaluation of drug properties and pharmacokinetics

Based on the provided pharmacological parameters and existing research, the preliminary evaluation of SG's pharmacological properties is as follows:

• absorb As a compound with high LogP and low water solubility, the oral absorption of SG may be limited by its dissolution rate. The glycosylation structure may make it more easily involved in active transport through glucose transporters (such as SGLT1) in small intestinal epithelial cells than glycosides, but overall bioavailability is expected to be low. Formulation strategies such as nanocrystals, solid dispersions, phospholipid complexes, cyclodextrin inclusion complexes are crucial for improving their solubility and absorption.
• distribution A high molecular weight (574.8) and moderate TPSA indicate low blood-brain barrier (BBB) permeability, which reduces the risk of central nervous system side effects but also limits its potential for use in central nervous system diseases. Their parental nature may result in a higher distribution of them in tissues rich in membrane structures, such as the liver, adrenal glands, and gonads.
• Metabolism As a sterol glycoside, SG may undergo two main metabolic pathways in the body: ① hydrolysis under the action of β - glucosidase in the gut microbiota, producing aglycone sitosterol and glucose, which can be absorbed and further metabolized (such as hydroxylation and side chain oxidation); ② It undergoes phase I (such as oxidation) and phase II (such as glucuronidation and sulfation) metabolism in the liver. The specific metabolic enzyme profile (such as CYP450 subtypes) remains to be clarified.
• excretion The prototype drug and its metabolites may be mainly excreted through bile and feces, with some water-soluble metabolites excreted through the kidneys.
• Preliminary Safety Assessment：
  • HERG inhibition The prediction is' no ', indicating that the risk of inducing QT interval prolongation and apical torsion ventricular tachycardia is low, and the cardiovascular safety is preliminarily optimistic.
  • Genotoxicity The predicted value of Ames test is 0.0, indicating no mutagenicity in this experimental system, but it needs to be confirmed through a complete combination of in vitro and in vivo genetic toxicity tests.
  • General toxicity Plant sterols and their glycosides, as food ingredients, have been shown to be relatively safe for human consumption over a long period of time. However, systematic research is still needed on the subchronic, chronic, and reproductive toxicity of high-dose medicinal use.

Clinical application prospects and prospects

SG, as a natural active molecule with multiple targets and effects, has broad development prospects in the following fields:

1. Hypolipidemic/antiatherosclerotic drugs or functional food additives This is the most direct application direction of SG. Can be developed as:
   • prescription drug As a supplement or alternative to statins, it is particularly suitable for patients with statin intolerance. It is necessary to solve its bioavailability problem through structural modification or advanced formulation technology, and conduct strict clinical trials to verify its efficacy and safety.
   • Over the Counter (OTC) or Health Food As an upgraded form of plant sterol esters, SG has a clear molecular structure and multi-target mechanism, which can be developed into a lipid-lowering health product with stronger scientific claims.
2. Adjuvant therapy for benign prostatic hyperplasia (BPH)Based on its 5 α - reductase inhibitory activity, SG can be used in combination with existing drugs (such as finasteride) or in the development of compound formulations for the treatment of BPH, which may have the advantage of minimal side effects of plant-based drugs.
3. Anti inflammatory adjuvant therapy Its anti-inflammatory mechanism is clear and can be used for auxiliary management of inflammation related chronic diseases, such as metabolic inflammation, fatty liver, etc.
4. cosmetic ingredient Its antioxidant, anti-inflammatory properties, and 5 α - reductase inhibitory activity can be used to develop cosmetics with anti-aging, soothing, and anti hair loss functions.

However, achieving its clinical application still requires overcoming the following challenges and conducting in-depth research:
* challenge① Poor water solubility and low bioavailability; ② The natural source content is limited, and the chemical or biological synthesis pathways need to be optimized to achieve large-scale production; ③ The multi-target characteristic is both advantageous and may bring unpredictable off target effects, requiring a more precise mechanism of action map.
* Future research directions：① structural optimization Modify the sugar moiety or steroid nucleus to enhance activity, solubility, and metabolic stability. ② Formulation innovation Develop new nano drug delivery systems (such as liposomes, polymer nanoparticles), self microemulsions, etc., to improve delivery efficiency. ③ Systematic pharmacokinetic study Clarify its ADME process in various animal models and humans. ④ In depth mechanism research Using proteomics, metabolomics, network pharmacology and other methods, comprehensively elucidate its functional network. ⑤ High quality clinical research Conduct randomized controlled clinical trials from phase I to phase III to confirm its effectiveness and safety in treating specific diseases such as mixed hyperlipidemia and mild BPH.

Conclusion

As a naturally occurring sterol glycoside, stigmasterol glucoside exhibits diverse pharmacological activities beyond traditional plant sterols due to its unique chemical structure. Especially in regulating lipid metabolism, it forms a synergistic network by acting on multiple key targets such as HMGCR, LDLR, PCSK9, CETP, PPARA, etc., providing a solid scientific basis for its use as a candidate drug for reducing blood lipids and anti atherosclerosis. Meanwhile, its combination of 5 α - reductase inhibition, anti-inflammatory, and antioxidant activities further expands its potential therapeutic applications. Despite the challenges of solubility and bioavailability in drug development, modern pharmaceutical chemical modifications and novel formulation technologies provide feasible solutions for this. In the future, through interdisciplinary in-depth research and development, sitosterol glucoside is expected to be transformed from a common plant component into a novel drug or highly effective functional factor for treating metabolic and chronic inflammatory diseases, contributing to human health through natural efforts.