Synonym name: Nevadensin; Ethoxychelerythrine; 152743-19-6
Catalogue No.: BP0906
Cas No.: 10176-66-6
Formula: C₁₈H₁₆O₇
Mol Weight: 344.319
Botanical Source:

Purity: 95%~99%
Analysis Method: HPLC-DAD or/and HPLC-ELSD
Identification Method: Mass, NMR
Packing: Brown vial or HDPE plastic bottle
Can be supplied from milligrams to grams.

For Reference Standard and R&D, Not for Human Use Directly.
Inquire for bulk scale.

​Description:

Nevadensin is an important herb-based constituent inhibiting estragole bioactivation. Nevadensin protects against a methyleugenol-induced marker of hepatocarcinogenicity in male F344 rat. It also exhibits inhibition activity against Mycobacterium tuberculosis, with equal MIC value of 200 microg/mL.

References:

Food Chem Toxicol. 2014 Dec;74:28-34.    

The natural basil flavonoid nevadensin protects against a methyleugenol-induced marker of hepatocarcinogenicity in male F344 rat.  

The alkenylbenzene methyleugenol occurs naturally in a variety of spices and herbs, including basil, and their essential oils. At high dose levels methyleugenol induces hepatocarcinogenicity in rodents following bioactivation to 1'-sulfooxymethyleugenol which forms DNA adducts. 

METHODS AND RESULTS:

This study investigated whether the inhibitory effect of the basil flavonoid Nevadensin on sulfotransferase (SULT)-mediated bioactivation of methyleugenol observed in vitro would also be reflected in a reduction of DNA adduct formation and a reduction in an early marker for liver carcinogenesis in an 8-week rat study. Co-exposure to methyleugenol and Nevadensin orally resulted in a significant inhibition of liver methyleugenol DNA adduct formation and in inhibition of hepatocellular altered foci induction, representing indicators for initiation of neoplasia. 

CONCLUSIONS:

These results suggest that tumor formation could be lower in rodent bioassays when methyleugenol would be dosed in a matrix containing SULT inhibitors such as Nevadensin compared to experiments using the pure methyleugenol.    

Arch Pharm Res. 2003 Oct;26(10):816-20.    

Antimycobacterial and antioxidant flavones from Limnophila geoffrayi.    

The chloroform extract of the aerial part of Limnophila geoffrayi showed antimycobacterial and antioxidant activities. 

METHODS AND RESULTS:

Bioassay-guided fractionation has led to the isolation of the flavones Nevadensin (5,7-dihydroxy-6,8,4'-trimethoxyflavone, 1) and isothymusin (6,7-dimethoxy-5,8,4'-trihydroxyflavone, 2). Both compounds 1 and 2 exhibited inhibition activity against Mycobacterium tuberculosis, with equal MIC value of 200 microg/mL. Only compound 2 exhibited antioxidant activity against the radical scavenging ability of DPPH, with the IC50 value of 7.7 microg/mL. 

CONCLUSIONS:

The crude hexane, chloroform and methanol extracts as well as the pure compounds 1 and 2 did not exhibit mutagenic activity in the Bacillus subtilis recassay.    

Toxicol Appl Pharmacol. 2010 Jun 1;245(2):179-90.    

Identification of nevadensin as an important herb-based constituent inhibiting estragole bioactivation and physiology-based biokinetic modeling of its possible in vivo effect.    

Estragole is a natural constituent of several herbs and spices including sweet basil. In rodent bioassays, estragole induces hepatomas, an effect ascribed to estragole bioactivation to 1'-sulfooxyestragole resulting in DNA adduct formation. 

METHODS AND RESULTS:

The present paper identifies Nevadensin as a basil constituent able to inhibit DNA adduct formation in rat hepatocytes exposed to the proximate carcinogen 1'-hydroxyestragole and Nevadensin. This inhibition occurs at the level of sulfotransferase (SULT)-mediated bioactivation of 1'-hydroxyestragole. The Ki for SULT inhibition by Nevadensin was 4 nM in male rat and human liver fractions. Furthermore, Nevadensin up to 20 microM did not inhibit 1'-hydroxyestragole detoxification by glucuronidation and oxidation. The inhibition of SULT by Nevadensin was incorporated into the recently developed physiologically based biokinetic (PBBK) rat and human models for estragole bioactivation and detoxification. The results predict that co-administration of estragole at a level inducing hepatic tumors in vivo (50mg/kg bw) with Nevadensin at a molar ratio of 0.06, representing the ratio of their occurrence in basil, results in almost 100% inhibition of the ultimate carcinogen 1'-sulfooxyestragole when assuming 100% uptake of Nevadensin. Assuming 1% uptake, inhibition would still amount to more than 83%. Altogether these data point at a Nevadensin-mediated inhibition of the formation of the ultimate carcinogenic metabolite of estragole, without reducing the capacity to detoxify 1'-hydroxyestragole via glucuronidation or oxidation. 

CONCLUSIONS:

These data also point at a potential reduction of the cancer risk when estragole exposure occurs within a food matrix containing SULT inhibitors compared to what is observed upon exposure to pure estragole.    

HPLC of Lysionotin

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

Introduction/Overview

Lysionotin (CAS No.: 10176-66-6) is a trimethoxyflavonoid natural product with unique structural characteristics. Due to its special arrangement of polyhydroxyl and methoxy substituents, it exhibits abundant biological activity, especially showing potential pharmacological value in the field of anti-tuberculosis. As a plant metabolite, spider-spider not only plays an important role in plant defense mechanisms, but its potential for drug development is also attracting increasing attention. In recent years, as the problem of tuberculosis drug resistance has worsened, finding novel anti-tuberculosis drugs has become an urgent task in the global public health sector. Stone spiderin has become one of the hot topics in natural product pharmacology research due to its action on key targets of Mycobacterium tuberculosis.

This paper will systematically review the chemical structure and physicochemical properties of spiderlingin, plant origin and extraction methods, pharmacological activity and mechanism of action, druggability evaluation and pharmacokinetic characteristics, and explore its clinical application prospects and future development directions, aiming to provide comprehensive reference materials for researchers in related fields.

Chemical structure and physicochemical properties

Cyclophyllin belongs to the flavonoid class, specifically trimethoxyflavone, with a molecular formula of C18H16O7 and a molecular weight of 344.3190. Its structural feature is that methoxy groups replace the 6, 8, and 4' positions of the flavonoid backbone, while hydroxyl groups replace the 5 and 7' positions, forming a unique functional group arrangement. This structure allows it to combine the hydrophobicity of methoxyflavones and the hydrophilicity of hydroxyflavones, giving it a solid bioactive foundation.

In terms of physicochemical properties, the LogP value of soralanin is 2.4315, indicating moderate lipid solubility that facilitates cell membrane penetration. The polar surface area (TPSA) is 98.36 Ų, indicating a certain degree of polarity, which facilitates binding to biological targets. Low water solubility (about 0.0108 mg/mL) suggests limited solubility in the aqueous phase, which may affect its bioavailability. The low permeability of the blood-brain barrier indicates limited distribution in the central nervous system. The hERG channel inhibition test was negative, indicating a low risk of cardiotoxicity. The Ames test value was 1.2, indicating a relatively low genotoxicity risk.

The molecular structure of lithosardines is shown in the figure below (the structural diagram should be inserted here). Its distribution of multiple hydroxyl and methoxy groups provides diverse binding sites for its biological activity, especially showing high affinity for interactions with protein targets.

Plant Origins and Extraction Methods

Lysionotus is mainly found in plants of the genus Lysionotus, with higher levels in the leaves and stems of species such as Lysionotus pauciflorus. These plants are widely distributed in temperate and subtropical regions of Asia and have traditionally been used as traditional Chinese medicinal herbs, with effects such as clearing heat and detoxifying, reducing inflammation, and relieving pain.

Common methods for extracting spiderin include solvent extraction, ultrasound-assisted extraction, and column chromatography separation. Ethanol or methanol is generally used as the extraction solvent, combined with ultrasonic-assisted technology to improve extraction efficiency. The extraction process usually is:

1. Sample pretreatment: Plant materials are dried and crushed, then screened evenly.
2. Solvent extraction: Use 70%-95% ethanol or methanol for extraction at room temperature up to 50°C, with a time of about 2-4 hours.
3. Concentration: Concentrate the extract by rotating the evaporator to remove the solvent.
4. Separation and purification: Separation and purification are performed using silica gel column chromatography or high-performance liquid chromatography (HPLC) to obtain high-purity spiderganil.
5. Structural identification: Confirm its structure using techniques such as nuclear magnetic resonance (NMR), mass spectrometry (MS), and infrared spectroscopy (IR).

In recent years, green extraction technologies such as supercritical CO2 extraction and microwave-assisted extraction have also been attempted to extract sphenolic spiderin to improve efficiency and reduce environmental pollution.

Pharmacological activity research

Research on the pharmacological activity of stenxorlanin mainly focuses on its anti-tuberculosis effects and related antibacterial and anti-inflammatory activities. Multiple in vitro and in vivo experiments have shown that spiderin has a significant inhibitory effect on Mycobacterium tuberculosis, especially showing good activity in drug-resistant strains.

Anti-tuberculosis activity

Cycloloxin interferes with the physiological functions of Mycobacterium tuberculosis through a multi-target mechanism. It inhibits key targets such as INHA (pyruvate dehydrogenase inhibitor protein), KATG (catalase), EMBR (cell wall synthase), RPOB (RNA polymerase β subunit), and PNC1 (penicillin-binding protein), blocking bacterial energy metabolism, cell wall synthesis, and gene transcription, ultimately leading to bacterial death or growth restriction.

Other pharmacological activities

In addition to its anti-tuberculosis properties, lithizolanin also exhibits certain antioxidant and anti-inflammatory activities. Its hydroxyl groups can effectively eliminate free radicals and reduce oxidative stress damage. In inflammation models, spiderfurin reduces tissue inflammatory responses by inhibiting the expression of inflammatory factors such as TNF-α and IL-6. Additionally, some studies suggest it may have potential effects in liver protection and neuroprotection, but the related mechanisms still require further exploration.

Mechanism of action and molecular targets

The mechanism of action of spiderin involves multiple biological pathways, mainly by binding to key enzymes and proteins of Mycobacterium tuberculosis, thereby inhibiting its biosynthesis and metabolic processes.

1. INHA (pyruvate dehydrogenase inhibitor protein): Pyruvate dehydrogenase inhibitory protein: Cyclogonyl binds to INHA, interfering with bacterial energy metabolism and reducing ATP production, leading to insufficient energy supply for bacteria.
2. KATG (catalase): As an important enzyme for bacteria to defend against oxidative stress, KATG is inhibited by cycloplasmin, reducing bacterial resistance to oxidative damage and making it more susceptible to attack by the host's immune system.
3. EMBR (Cell Wall Synthase): Stone spiderin blocks cell wall synthesis, disrupts the structural integrity of bacterial cells, and leads to bacterial lysis.
4. RPOB (RNA Polymerase β Subunit): By inhibiting RNA polymerase, spiderlangin blocks bacterial gene transcription and suppresses protein synthesis.
5. PNC1 (penicillin-binding protein): affects the cross-linking process of bacterial cell walls, enhancing antibacterial effects.

Molecular docking and kinetic simulations show that the methoxy and hydroxyl groups of soracloflin form stable binding to the active site of the target protein through hydrogen bonding and hydrophobic interactions, enhancing its inhibitory activity. Additionally, the structural characteristics of soragonal allow it to cross bacterial cell walls and reach target sites.

Druggability evaluation and pharmacokinetics

The drug-producing parameters of lithizolanin indicate that it has certain development potential:

• The molecular weight (344.3190) complies with the Lipinski rule, which is beneficial for oral absorption.
• The LogP value (2.4315) is moderate, balancing lipophilic and water solubility, which facilitates cell membrane penetration.
• TPSA (98.36 Ų) is suitable for binding to multiple targets without affecting biofilm penetration.
• Low water solubility (0.0108 mg/mL) may limit its oral bioavailability and requires pharmaceutical improvement.
• The low permeability of the blood-brain barrier suggests a lower risk of side effects in the central nervous system.
• hERG channel inhibition negative, reducing the risk of cardiotoxicity.
• Ames test results (1.2) showed a low genotoxicity risk.

In terms of pharmacokinetics, current research is relatively limited. Preliminary in vivo experiments show that lithosarin is absorbed slowly after oral administration, has a moderate plasma half-life, and is mainly metabolized by the liver, though the metabolites are not yet fully identified. Due to its low water solubility and limited bioavailability, future technologies such as nanocarriers, liposomes, or solid dispersions will need to improve its stability and absorption efficiency in vivo.

Prospects and outlooks for clinical applications

As a multi-target natural anti-tuberculosis product, spiderin has good pharmacological activity and safety, demonstrating potential as a novel anti-tuberculosis drug. Its multi-target mechanism helps overcome the tendency of single-target drugs to develop resistance, providing new ideas for tuberculosis treatment.

The key to future clinical applications lies in:

1. Optimized formulation: Improves water solubility and bioavailability to ensure effective concentration in the body.
2. Systematic pharmacokinetic studies: clarify its distribution, metabolic pathways, and excretion mechanisms in vivo.
3. Safety evaluation: Conduct long-term toxicology and mutagenicity studies to ensure clinical safety.
4. Combination therapy research: Exploring synergies with existing anti-tuberculosis drugs to reduce resistance risk.
5. Clinical trial design: Gradually advance clinical trials from in vitro and animal models to human to verify efficacy and safety.

In addition, the potential applications of lithosarin in anti-inflammatory and antioxidant areas are worth further exploration, and may play an adjunctive therapeutic role in various chronic diseases.

Conclusion

As a naturally occurring trimethoxyflavonoid with a unique structure, spiderling has become an important subject for pharmacological research of natural products due to its remarkable anti-tuberculosis activity and excellent druggability parameters. Its multi-target mechanism of action offers new directions for the development of anti-tuberculosis drugs. Although research into its pharmacokinetics and clinical applications is still in its early stages, with the development of extraction and purification technologies and drug delivery systems, cyclospiderlin is expected to become a novel candidate drug for future anti-tuberculosis treatments.

Future research should focus on deeply elucidating its mechanism of action, optimizing drug formulations, and conducting systematic preclinical and clinical studies, aiming to move cyclocytosinol from the laboratory to clinical application, benefiting tuberculosis patients while expanding its potential in other disease fields.