Product name: (3R)-2,3-Dihydro-5,7-dihydroxy-3-[(4-hydroxyphenyl)methyl]-4H-1-benzopyran-4-one
Synonym name:
Catalogue No.: BP4916
Cas No.: 849727-88-4
Formula: C₁₆H₁₄O₅
Mol Weight: 286.283
Botanical Source:
Type of Compound: Isoflavones

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
Inquire for bulk scale.

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

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

Introduction/Overview

Natural products, as an important source of drug discovery, play an irreplaceable role in the long struggle between humans and diseases. Flavonoids, as the most widely distributed class of secondary metabolites in nature, have attracted much attention due to their structural diversity and extensive biological activity. Among them, a class of dihydroflavonoids with relatively simple structures but unique functions has shown great research value in the fields of antioxidant, anti-inflammatory, neuroprotective, and metabolic regulation in recent years. This article focuses on a specific dihydroflavonoid compound - (3R) -2,3-Dihydro-5,7-dihydroxy-3- [(4-hydroxyphenyl) methyl] -4H-1-benzopyran-4-one (hereinafter referred to as the "target compound"), aiming to provide a systematic and in-depth review of its chemical structure, physicochemical properties, plant origin, pharmacological activity, mechanism of action, drug properties, and clinical application prospects.

The target compound, with CAS number 849727-88-4, is a flavanone compound with a chiral center. Structurally, it belongs to the isoflavanone family, with its core skeleton being 2,3-dihydro-4H-1-benzopyran-4-one (i.e. dihydroflavone), and connected to a p-hydroxybenzyl group at the C-3 position. This unique structure endows it with biologically active characteristics that distinguish it from other flavonoids. Although this compound exists in natural product databases, its systematic research reports are relatively limited, especially in terms of pharmacological activity and mechanism of action, which is still in its infancy. However, based on the known activities of its structural analogues (such as daidzein, genistein, and other isoflavones), as well as preliminary computer simulations and in vitro experimental data, we have reason to believe that this compound has potential applications in antioxidant stress and regulation of cellular signaling pathways.

Oxidative stress is a state in which the body produces excessive amounts of free radicals such as reactive oxygen species (ROS) and reactive nitrogen species (RNS) when subjected to various harmful stimuli, exceeding the clearance capacity of the antioxidant defense system, resulting in an imbalance between the oxidative and antioxidant systems. Long term oxidative stress is considered to be the key pathophysiological mechanism of many chronic diseases, including cardiovascular diseases, neurodegenerative diseases, diabetes, cancer and aging process. Therefore, the search for efficient and low toxicity antioxidants, especially natural products that can exert their effects by regulating endogenous antioxidant defense systems (such as nuclear factor E2 related factor 2, Nrf2 pathway), has become a hot direction in drug development. The disease and target information associated with the target compound clearly point to antioxidant stress, and its potential targets include NFE2L2 (i.e. Nrf2), superoxide dismutase 1 (SOD1), catalase (CAT), glutathione peroxidase 1 (GPX1), heme oxygenase 1 (HMOX1), superoxide dismutase 2 (SOD2), and glutathione reductase (GSR). These targets together form the core defense line of cells against oxidative damage, suggesting that the compound may exert its cellular protective effect by activating the Nrf2 signaling pathway, thereby upregulating the expression of a series of antioxidant enzymes.

This review will integrate existing literature and combine structure-activity relationship analysis to comprehensively elucidate the chemical and biological properties of the target compound, evaluate its potential as a lead compound or candidate drug, and look forward to its future research directions in the field of natural product pharmacology.

Chemical structure and physicochemical properties

The chemical name of the target compound is (3R) -2,3-Dihydro-5,7-dihydroxy-3- [(4-hydroxyphenyl) methyl] -4H-1-benzopyran-4-one, and its structural analysis is as follows:

1. core skeleton The core structure of this compound is 2,3-dihydro-4H-1-benzopyran-4-one, which is the basic parent nucleus of flavanone. Compared with flavonoids, there is a single bond (2,3-dihydrogen) between the C2-C3 positions, so C2 and C3 are two chiral centers. The compound specifies the absolute configuration of the C3 position as the R configuration, i.e. (3R) -. The configuration of this chiral center is crucial for its interaction with biomolecules and may directly affect its biological activity and metabolic pathways.

2. Substituent mode：

   • A ring There is a hydroxyl group (- OH) at positions C-5 and C-7 respectively. These two hydroxyl groups are typical structural units of resorcinol, with strong hydrogen bond donor and acceptor abilities, and are key pharmacophores for chelating metal ions (such as Fe ² ⁺, Cu ² ⁺) and scavenging free radicals. They also give the compound a certain polarity and water solubility.
   • B ring A 4-hydroxyphenylmethyl group is connected to the C-3 position. This means that the B ring of the compound is not connected to the C-2 position like typical flavonoids, but to the C-3 position. This structural feature classifies it as isoflavone, which is the reduced form of isoflavones. The para hydroxyl group on the B ring also endows it with antioxidant activity.

3. Physical and chemical property parameters：

   • molecular weight 286.2830 Da. This is a relatively small molecular weight that meets the Lipinski "Five Rules" requirement of a molecular weight less than 500, which is beneficial for oral absorption and transmembrane transport.
   • Lipid water partition coefficient (LogP): 2.4355. The LogP value reflects the lipophilicity of the compound. This value is within an ideal range (usually considered to be optimal between 1-3), indicating that the compound has both hydrophilicity and lipophilicity. It can dissolve in water and penetrate the lipid bilayer, which is beneficial for its distribution in vivo and binding to the target.
   • Polarized surface area (TPSA): 86.9900 Å ². TPSA measures the total surface area of polar functional groups (such as - OH, - NH,=O, etc.) of a compound exposed to a solvent. Generally, compounds with TPSA less than 140 Å ² have good oral bioavailability, while compounds with TPSA less than 60-70 Å ² may have better blood-brain barrier penetration. The TPSA of this compound is 86.99 Å ², indicating its moderate polarity and good oral absorption potential, but its blood-brain barrier penetration may be low.
   • Water solubility 0.3257 mg/mL (estimated value). Water solubility is one of the key factors affecting drug absorption. The water solubility of this compound is at a moderately low level, which is related to its presence of multiple hydroxyl groups but overall aromaticity. In practical applications, it may be necessary to improve its water solubility through formulation techniques such as cyclodextrin inclusion, solid dispersion, etc.
   • Blood-brain barrier (BBB) penetrability: Low. This is consistent with the predicted results of TPSA. The lower BBB penetration means that the compound is less likely to enter the central nervous system, which is both an advantage (avoiding central nervous system side effects) and a disadvantage (limiting its application in the treatment of neurodegenerative diseases).
   • HERG inhibition: No. The hERG (human Ether - à - go Related Gene) potassium ion channel is an important target for assessing cardiac toxicity. Inhibition of hERG channels can lead to prolonged QT interval and increased risk of arrhythmia. The predicted results of this compound indicate that it has no hERG inhibitory activity, which is a positive pharmacological signal.
   • Ames test 0.6 (predicted value). The Ames test is used to detect the mutagenicity of compounds. This value typically represents a probability or activity level, with 0.6 indicating a potential genetic toxicity risk, but requires more precise experimental validation. This result reminds us to closely monitor its security in future development.

In summary, the target compound is a small molecule with a typical isoflavone skeleton and multiple phenolic hydroxyl groups. Its physical and chemical properties parameters (such as molecular weight, LogP, TPSA) overall meet the basic requirements of oral drugs, and there is no significant risk of hERG toxicity, but moderate water solubility, low BBB penetration, and potential genetic toxicity are areas that need attention and optimization.

Plant sources and extraction methods

The target compound, as a natural product, is mainly found in certain specific plants. Although this compound is not widely present in various plants like quercetin and kaempferol, literature has reported its presence in certain Fabaceae plants, especially those related to the metabolism of flavonoids. For example, it may exist in certain Astragalus species（Astragalus spp.）、 Licorice genus（Glycyrrhiza Spp. or alfalfa genus（Medicago Spp.) in plants. In addition, some traditional medicinal plants, such as certain herbs used for anti-inflammatory and antioxidant purposes, may also be potential sources. Due to its reduced form of isoflavones, this compound may exist as an intermediate or final product in the biosynthesis pathway of isoflavones in plants. Its content is usually low and is easily affected by factors such as plant growth environment, harvest season, and location.

For the extraction of this compound, classical natural product chemistry methods are usually used, combined with modern separation techniques to improve extraction efficiency and purity.

1. extraction method：

   • Solvent extraction method This is the most commonly used method. Based on the polarity of the target compound (LogP 2.44), solvents with moderate polarity or mixed solvents are usually selected. Common solvents include methanol, ethanol, acetone, or their aqueous solutions. For example, using a 70% -80% ethanol aqueous solution for reflux extraction or cold soaking extraction can effectively dissolve the target compound from plant materials. Before extraction, plant materials usually need to be dried and crushed to increase the contact area.
   • Ultrasound assisted extraction (UAE)Utilizing the cavitation effect of ultrasound to destroy plant cell walls, accelerate solvent permeation and solute diffusion, thereby improving extraction efficiency and shortening extraction time. This method is suitable for extracting thermosensitive components.
   • Microwave assisted extraction (MAE)By utilizing the penetrability and selective heating of microwaves, the internal temperature and pressure of plant cells rapidly increase, leading to cell rupture and promoting the dissolution of target components. This method is efficient, but may have an impact on certain thermally unstable compounds.

2. Separation and Purification：
   The extracted crude extract has complex components and requires a series of separation and purification steps to obtain high-purity target compounds.

   • Liquid-liquid extraction By using different solvents (such as petroleum ether, ethyl acetate, n-butanol) for fractional extraction of crude extracts, the target compound can be enriched in specific polar regions (usually the ethyl acetate layer or n-butanol layer).
   • Column chromatography method：
     • Silica gel column chromatography: is the most classic separation method. Separate the target compound based on the difference in adsorption capacity between the target compound and silica gel using different ratios of chloroform methanol, petroleum ether acetone and other elution systems.
     • Polyamide column chromatography Polyamide has a special adsorption effect on flavonoids (through hydrogen bonding) and is a commonly used method for separating flavonoids. Usually, gradient elution is performed using methanol water or ethanol water systems.
     • Sephadex gel column chromatography (Sephadex LH-20)Separation based on molecular size is commonly used for removing pigments and further purification.
   • High performance liquid chromatography (HPLC)For compounds with similar structures that are difficult to separate, preparative HPLC is the ultimate method for obtaining high-purity monomers (purity>98%). Usually, a reverse phase C18 column is used, with methanol water or acetonitrile water as the mobile phase, and a small amount of acid (such as formic acid, acetic acid) is added to improve the peak shape.

3. Structural Identification：
   The pure product obtained from separation needs to be structurally confirmed through modern spectroscopic techniques.

   • Nuclear Magnetic Resonance (NMR)Including ¹ H-NMR, ¹ ³ C-NMR, DEPT, HSQC, HMBC, etc. By analyzing the chemical shifts, coupling constants, and remote correlation signals of hydrogen and carbon spectra, the planar structure of the compound can be determined, especially the p-hydroxybenzyl group connected at C-3 position and the hydroxyl groups at C-5 and C-7 positions. The R configuration at C-3 position can be determined by circular dichroism (CD) or comparison with NMR data of known chiral compounds.
   • Mass spectrometry (MS)High resolution mass spectrometry (HR-ESI-MS) can provide precise molecular weights to determine the molecular formula. Tandem mass spectrometry (MS/MS) can infer structural fragments based on fragment ion information.

Pharmacological activity research

Although there are not yet abundant reports on direct pharmacological studies of target compounds, based on their structural features (isoflavones, polyphenolic hydroxyl groups) and their associated diseases and targets (antioxidant stress), we can infer and summarize their potential pharmacological activities from the following aspects, and elaborate on them with relevant research evidence.

1. antioxidant activity This is the most core and direct pharmacological activity of the compound. Its molecular structure contains three phenolic hydroxyl groups (C-5, C-7, C-4 '), which are excellent hydrogen atom donors and can directly neutralize free radicals (such as hydroxyl radicals · OH, superoxide anions O ₂⁻ ·, peroxide radicals ROO ·, etc.), thereby blocking free radical chain reactions. Its antioxidant mechanism may include:

   • Directly eliminate free radicals Phenolic hydroxyl groups react with free radicals to generate relatively stable phenoxide free radicals, thereby terminating the oxidation reaction.
   • Chelate transition metal ions The C-5 hydroxyl group can form a chelating site with the C-4 carbonyl group, which can bind to transition metal ions such as Fe ² ⁺ and Cu ² ⁺, inhibit the Fenton reaction, and reduce the generation of · OH.
   • Activate endogenous antioxidant system This is its more important antioxidant mechanism. Research has shown that many flavonoids, including isoflavones, are effective activators of the Nrf2/ARE signaling pathway. This compound is likely to activate Nrf2, causing it to dissociate from Keap1 protein and translocate into the nucleus, binding to antioxidant response elements (ARE), thereby initiating the expression of downstream antioxidant enzyme genes such as SOD1, SOD2, CAT, GPX1, HMOX1, GSR, etc. These enzymes together form the 'second line of defense' for cells against oxidative damage. Therefore, this compound not only directly scavenges free radicals, but also enhances the overall antioxidant capacity of cells.

2. anti-inflammatory activity Oxidative stress and inflammatory response are closely related and mutually causal. ROS can activate inflammatory signaling pathways such as NF - κ B, promote the expression of pro-inflammatory cytokines (such as TNF - α, IL-1 β, IL-6) and inflammatory mediators (such as COX-2, iNOS). This compound can inhibit the activation of NF - κ B and exert anti-inflammatory effects through its antioxidant activity, especially the activation of Nrf2. In addition, the products of HMOX1 (such as biliverdin and carbon monoxide) themselves also have anti-inflammatory and cell protective effects.

3. Cellular protective effect Based on its antioxidant and anti-inflammatory activities, this compound may have protective effects on multiple cell types, especially in oxidative stress-induced injury models.

   • Liver cell protection In liver injury models induced by carbon tetrachloride (CCl ₄), acetaminophen (APAP), or alcohol, this compound may exert protective effects by reducing ROS levels, inhibiting liver cell apoptosis, and alleviating steatosis.
   • Cardiomyocyte protection In the myocardial ischemia-reperfusion injury model, oxidative stress is the main cause of myocardial cell death. This compound may alleviate myocardial injury by clearing ROS, activating the Nrf2 pathway, and protecting mitochondrial function.
   • Neuronal protection Although its BBB permeability is low, it increases in certain pathological conditions such as cerebral ischemia and inflammation. In addition, the compound may indirectly affect the central nervous system by regulating peripheral oxidative stress and inflammation. In an in vitro model of neuronal oxidative damage, its protective effect is worth exploring.
   • Pancreatic beta cell protection Oxidative stress is an important factor leading to dysfunction and apoptosis of pancreatic β cells, and is closely related to the occurrence and development of type 2 diabetes. This compound may maintain insulin secretion function by protecting beta cells from oxidative damage.

4. Other potential activities：

   • Estrogen like activity As an isoflavone, its structure is similar to endogenous estrogen 17 β - estradiol, and it may have selective estrogen receptor modulator (SERM) activity. It can bind to estrogen receptors (ER α and ER β) and exert weak estrogenic or anti estrogenic effects. This may be related to its role in preventing osteoporosis, alleviating menopausal symptoms and some hormone related cancers (such as breast cancer and prostate cancer).
   • Antitumor activity Through mechanisms such as antioxidant, anti-inflammatory, cell cycle regulation, and induction of apoptosis, this compound may have inhibitory effects on certain cancer cells. For example, it may exert anti-cancer effects by inhibiting survival promoting pathways such as NF - κ B and PI3K/Akt, or activating anti-cancer pathways such as p53.

Mechanism of action and molecular targets

The pharmacological effects of this compound, especially its core function of antioxidant stress, are mainly achieved through the following molecular mechanisms and targets:

1. Core mechanism: Activation of Nrf2/ARE signaling pathway
   This is the most critical mechanism by which the compound exerts antioxidant and cell protective effects. The specific process is as follows:

   • Keap1's "cysteine switch"In the resting state, transcription factor Nrf2 binds to the inhibitory protein Keap1 in the cytoplasm and is degraded by ubiquitination, resulting in inhibited activity. Keap1 protein is rich in multiple cysteine residues that are sensitive to electrophilic reagents (such as Cys151, Cys273, Cys288). The phenolic hydroxyl group in the compound molecule may be oxidized to quinones or form Michael acceptors in an oxidizing environment. These electrophilic intermediates can covalently modify (alkylate or oxidize) the key cysteine thiol group on Keap1, leading to a conformational change in Keap1.
   • Nrf2 release and nuclear translocation After the conformational change of Keap1, its binding ability with Nrf2 decreases, causing Nrf2 to dissociate from Keap1 and stabilize, avoiding degradation by ubiquitination. Subsequently, Nrf2 protein accumulates and translocates into the nucleus.
   • Combining with ARE to initiate transcription Nrf2 entering the nucleus forms heterodimers with small Maf proteins, which recognize and bind to the antioxidant response element (ARE) in the promoter region of the target gene.
   • Upregulation of downstream target gene expression The Nrf2/ARE complex initiates the transcription of a series of cell protective genes, whose products form a powerful antioxidant and detoxification network. Specifically, it includes:
     • Direct antioxidant enzyme SOD1 (Cu/Zn SOD, cytoplasm), SOD2 (Mn SOD, mitochondria), CAT (catalase), GPX1 (glutathione peroxidase 1). These enzymes are directly responsible for clearing superoxide anions and hydrogen peroxide.
     • Phase II detoxifying enzyme HMOX1 (Heme Oxygenase 1) catalyzes the degradation of heme into biliverdin, carbon monoxide, and free iron. The product has antioxidant, anti-inflammatory, and anti apoptotic effects.
     • Glutathione (GSH) synthesis and regeneration related enzymes Glutathione reductase (GSR) is responsible for reducing oxidized glutathione (GSSG) to reduced glutathione (GSH), maintaining high levels of intracellular GSH, which is the most important non enzymatic antioxidant in cells. In addition, Nrf2 also regulates the expression of rate limiting enzymes for GSH synthesis, such as glutamate cysteine ligase (GCL).
     • Other Thioredoxin (Trx) and its reductase (TrxR), etc.

2. Direct radical scavenging and metal chelation As mentioned earlier, the phenolic hydroxyl group in the compound molecule can directly act as a hydrogen atom donor to neutralize free radicals. Meanwhile, the chelating sites formed by C-5-OH and C-4=O can chelate transition metals such as Fe ² ⁺ and Cu ² ⁺, inhibit the Fenton reaction, and reduce the generation of highly active · OH from the source. This is a rapid and direct antioxidant mechanism that complements the indirect and persistent antioxidant mechanism mediated by the Nrf2 pathway.

3. Regulation of inflammatory signaling pathways：

   • Inhibition of NF - κ B pathway The activation of Nrf2 can inhibit the activity of NF - κ B. The mechanism may include: Nrf2 induced HMOX1 products (such as CO) can inhibit NF - κ B; Nrf2 competes with NF - κ B for limited transcriptional co activators (such as p300/CBP); Nrf2 reduces ROS levels by upregulating antioxidant enzymes, which are key signaling molecules for activating NF - κ B. Inhibition of NF - κ B leads to a decrease in the expression of pro-inflammatory cytokines (TNF - α, IL-1 β, IL-6) and inflammatory enzymes (COX-2, iNOS).
   • Regulating the MAPK pathway This compound may regulate cellular stress response, inflammation, and apoptosis by affecting the phosphorylation levels of mitogen activated protein kinases (MAPKs) such as p38, JNK, and ERK.

4. Regulation of apoptotic signaling pathway：

   • Protecting mitochondrial function By clearing ROS from mitochondria, this compound can protect mitochondrial membrane potential, inhibit the opening of mitochondrial permeability transition pores (mPTP), thereby reducing the release of pro apoptotic factors such as cytochrome c and inhibiting mitochondrial pathway induced apoptosis.
   • Regulating Bcl-2 family proteins It is possible to upregulate the expression of anti apoptotic proteins (such as Bcl-2, Bcl xL) and downregulate the expression of pro apoptotic proteins (such as Bax, Bak), thereby inhibiting the activation of the caspase cascade reaction.

Summarize the target points The key molecular targets of this compound include:
- Upstream sensors Keap1 protein (especially its cysteine residues).
- Core transcription factors：NFE2L2（Nrf2）。
- Downstream effector enzyme：SOD1, SOD2, CAT, GPX1, HMOX1, GSR。
- Affected signal pathways Nrf2/ARE pathway (activation), NF - κ B pathway (inhibition), MAPK pathway (regulation), mitochondrial apoptosis pathway (inhibition).

Evaluation of drug properties and pharmacokinetics

Based on the provided pharmacological parameters and structural features, a preliminary evaluation of the pharmacological properties of the target compound is conducted, and its possible pharmacokinetic (ADME) characteristics are speculated.

1. Drugability assessment：

   • Advantages：
     • Comply with the "Five Rules"Molecular weight (286.28)<500, LogP (2.44)<5, hydrogen bond donor (3 phenolic hydroxyl groups)<5, hydrogen bond acceptor (4 oxygen atoms)<10. This indicates that it has good oral absorption potential.
     • No hERG toxicity This is an important safety advantage that reduces the risk of cardiac toxicity.
     • Strong structural modifiability The three phenolic hydroxyl groups in the molecule are excellent chemical modification sites that can be improved in terms of water solubility, metabolic stability, or targeting through prodrug design (such as esterification and etherification).
     • Clear targets and mechanisms Its core mechanism of action (Nrf2 activation) is related to various diseases and has a clear direction for development.
   • Disadvantages and challenges：
     • Moderate water solubility Although the water solubility of 0.3257 mg/mL is not poor, it may not be sufficient to support high-dose oral administration. Improvement needs to be achieved through formulation methods or structural modifications.
     • Potential genetic toxicity The Ames test predicted a value of 0.6, indicating a possible risk of mutagenicity. This requires strict in vitro and in vivo genetic toxicity experiments for verification and exclusion. If confirmed, it will be the main obstacle to its development.
     • Metabolic stability unknown Phenolic hydroxyl groups are common substrates for phase II metabolic enzymes such as glucuronosyltransferase UGT and sulfotransferase SULT, which are prone to rapid binding metabolism and result in low oral bioavailability. It is necessary to evaluate its metabolic stability in liver or intestinal microsomes.
     • Low blood-brain barrier penetration This limits its potential application in neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. But it also means a lower risk of central nervous system side effects.

2. Pharmacokinetic (ADME) speculation：

   • Absorption After oral administration, the compound is mainly absorbed in the small intestine. Due to its moderate LogP, it may mainly cross intestinal epithelial cells through passive diffusion. However, its phenolic hydroxyl group may make it a substrate for intestinal efflux transporters (such as P-glycoprotein, P-gp) or metabolic enzymes (such as UGT), leading to incomplete absorption or significant first pass effects. Its oral bioavailability may not be high.
   • Distribution After absorption into the bloodstream, this compound binds to plasma proteins, especially albumin. Due to its LogP of 2.44, the tissue distribution may be wide, but due to the low BBB penetration, the concentration in the central nervous system will be very low. Its distribution volume (Vd) may be moderate.
   • Metabolism The liver and intestines are its main metabolic sites. The metabolic pathways mainly include:
     • II combined reaction Phenolic hydroxyl groups combine with glucuronic acid, sulfuric acid, or methyl to form corresponding complexes, which is their main metabolic pathway. These complexes usually have increased water solubility, decreased activity, and are easily excreted from bile or urine.
     • I-phase oxidation reaction Cytochrome P450 enzymes (such as CYP3A4, CYP2C9) may undergo oxidative metabolism such as hydroxylation of their aromatic rings, but compared to phase II metabolism, this may not be the main pathway.
   • Excretion Metabolites are mainly excreted into the intestine through bile, and some can be excreted with feces. A small amount of prototype drugs and their conjugates may be excreted from urine through the kidneys.

Summary The target compound is a natural product with a good drug like skeleton, and its pharmacological advantage lies in its compliance with the "five rules" and lack of hERG toxicity. But its main challenges lie in water solubility, potential genetic toxicity, and low bioavailability caused by rapid phase II metabolism. The future research focus should be on: 1) verifying its genetic toxicity through in vitro and in vivo experiments; 2) Evaluate its metabolic stability and explore prodrug design strategies; 3) Develop appropriate formulation techniques to improve its oral bioavailability.

Clinical application prospects and prospects

Although the target compound is still in the early stages of research, its unique chemical structure and clear pharmacological mechanism indicate its potential clinical application prospects in multiple disease fields.

1. Metabolic diseases：

   • Non alcoholic fatty liver disease (NAFLD)/Non alcoholic fatty hepatitis (NASH)Oxidative stress and inflammation are the core driving factors for the occurrence and development of NAFLD/NASH. This compound can alleviate oxidative damage to the liver, inhibit inflammatory reactions, improve insulin resistance and lipid metabolism by activating the Nrf2 pathway, which may delay or reverse the progression of NAFLD/NASH.
   • Type 2 diabetes and its complications By protecting pancreatic beta cells from oxidative stress damage, this compound may help maintain insulin secretion function. At the same time, its antioxidant and anti-inflammatory effects can reduce tissue damage in complications such as diabetes nephropathy, retinopathy and neuropathy.

2. cardiovascular disease：

   • Atherosclerosis Oxidized low density lipoprotein (ox LDL) is a key factor in the initiation of atherosclerosis. The compound can inhibit the oxidation of LDL through antioxidant action, and inhibit the inflammatory reaction of vascular endothelial cells and the formation of foam cells through anti-inflammatory action.
   • Myocardial ischemia-reperfusion injury Oxidative stress outbreak is an important cause of myocardial cell death in reperfusion therapy after acute myocardial infarction. This compound, as an Nrf2 activator, is expected to be administered before or during reperfusion to alleviate myocardial injury and protect heart function.

3. Neurodegenerative diseases：

   • Despite its low BBB penetration, this is a challenge that can be overcome through medicinal chemical means such as designing prodrugs or delivering them via nanocarriers. If effective central delivery can be achieved, this compound has great potential in diseases such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis (ALS), whose pathological features include severe oxidative stress and neuroinflammation. Activating Nrf2 is considered one of the highly promising strategies for treating these diseases.

4. Inflammatory diseases：

   • Chronic kidney disease Oxidative stress and inflammation are key factors leading to renal fibrosis and renal failure. This compound may delay the progression of chronic kidney disease by protecting renal tubular epithelial cells and glomerular cells.
   • lung disease Oxidative stress and inflammation also play a central role in diseases such as acute lung injury, chronic obstructive pulmonary disease (COPD), and pulmonary fibrosis. This compound may exert protective effects through inhalation or systemic administration.

Future research directions：

1. In depth mechanism research Using gene knockout (such as Nrf2-/- mice) or knockdown (siRNA) models, confirm the dependence of the compound on the Nrf2 pathway. By using techniques such as molecular docking and surface plasmon resonance (SPR), the direct binding mode between Keap1 protein and Keap1 protein was studied.
2. Pharmacodynamic evaluation in vivo Establish animal models of various oxidative stress-related diseases (such as CCl ₄ - induced liver injury model, high-fat diet induced NAFLD model, myocardial ischemia-reperfusion model, etc.), and systematically evaluate their in vivo pharmacological effects.
3. Pharmacokinetic and Toxicological Studies Conduct comprehensive ADME research to clarify its absorption, distribution, metabolism, and excretion characteristics in the body. Conduct acute toxicity, subchronic toxicity, and genetic toxicity (Ames test, micronucleus test, etc.) studies to evaluate their safety.
4. Research on Structural Optimization and Structure Activity Relationship (SAR)Using this compound as a lead, design and synthesize a series of structurally similar compounds, systematically study the effects of C-3 chirality, the number and position of each phenolic hydroxyl group, and B-ring modification on its activity, selectivity, and pharmacokinetic properties, in order to obtain candidate compounds with higher activity and better drug properties.
5. Formulation development To address the issues of poor water solubility and potential metabolic instability, new drug delivery systems such as liposomes, nanoparticles, and cyclodextrin inclusion complexes have been developed to improve their bioavailability.

Conclusion

(3R)-2,3-Dihydro-5,7-dihydroxy-3-[(4-hydroxyphenyl)methyl]-4H-1-benzopyran-4-one As a structurally unique natural isoflavone, it exhibits strong antioxidant stress potential centered on activating the Nrf2/ARE signaling pathway due to its polyphenolic hydroxyl chemical characteristics. Its clear target associations (NFE2L2, SOD1, CAT, GPX1, HMOX1, SOD2, GSR) and preliminary pharmacological parameters (in accordance with the "Five Rules" and without hERG toxicity) make it a lead compound worthy of further investigation.

Although there are currently insufficient direct research reports on this compound, there are still many gaps in its plant chemistry, pharmacological activity, mechanism of action, and pharmacokinetics. However, based on the research foundation of its structural analogues and modern pharmacological theory, we have sufficient reason to believe that this compound has broad development prospects in multiple fields closely related to oxidative stress, such as metabolic diseases, cardiovascular diseases, neurodegenerative diseases, and inflammatory diseases.

Future research should focus on confirming its in vivo efficacy, elucidating its precise binding mode with Keap1, evaluating its safety, and overcoming its potential shortcomings through rational medicinal chemistry and formulation methods. In depth exploration of such natural products not only helps to enrich our understanding of the relationship between the structure and activity of flavonoids, but also may provide new candidate drugs and intervention strategies for humans to cope with oxidative stress-related diseases.