BiochaninA

• Antioxidant activity
• Anti-inflammatory effects
• Hormonal balance
• Neuroprotective properties

• Bone health maintenance
• Cardiovascular health support
• Metabolic health improvement
• Cancer preventive potential
• Antimicrobial activity

Biochanin A (5,7-dihydroxy-4′-methoxyisoflavone) is a naturally occurring O-methylated isoflavone that exerts its biological effects through multiple mechanisms. As a phytoestrogen, Biochanin A’s primary mechanism involves interaction with estrogen receptors (ERs) due to its structural similarity to 17-β-estradiol. It demonstrates selective binding affinity, preferentially interacting with estrogen receptor-beta (ER-β) over estrogen receptor-alpha (ER-α), though with lower affinity than genistein. This selective receptor binding contributes to tissue-specific effects, as ER-β and ER-α are distributed differently throughout body tissues.

In tissues where ER-β predominates (such as bone, brain, vascular epithelium), Biochanin A typically exerts agonistic (estrogenic) effects. In tissues where ER-α is more prevalent (such as breast and uterine tissue), it may act as a partial antagonist, potentially inhibiting estrogen’s proliferative effects. Beyond its estrogenic activities, Biochanin A serves as a natural precursor to genistein through demethylation by cytochrome P450 enzymes, particularly CYP1A1 and CYP1B1. This conversion is significant because genistein possesses stronger estrogenic and tyrosine kinase inhibitory activities.

Biochanin A demonstrates significant antioxidant capabilities through multiple mechanisms. It directly neutralizes reactive oxygen species (ROS), particularly superoxide anions and hydroxyl radicals, through its phenolic hydroxyl groups. It also upregulates endogenous antioxidant enzymes including superoxide dismutase, catalase, and glutathione peroxidase through activation of the Nrf2 (Nuclear factor erythroid 2-related factor 2) pathway. Additionally, Biochanin A exhibits potent anti-inflammatory properties by inhibiting pro-inflammatory enzymes such as cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS), and by suppressing nuclear factor-kappa B (NF-κB) activation, thereby reducing the production of inflammatory cytokines.

This anti-inflammatory activity is partially mediated through peroxisome proliferator-activated receptor gamma (PPARγ) activation. In neuroinflammatory conditions, Biochanin A inhibits microglial activation and reduces the production of pro-inflammatory mediators through suppression of the TLRs/TIRAP/MyD88/NF-κB pathway. It also protects against neuronal apoptosis by modulating mitochondrial function and inhibiting caspase activation. Biochanin A influences cell cycle regulation and apoptosis pathways, which contributes to its potential anticancer effects.

It can induce cell cycle arrest at G2/M phase by modulating the expression of cyclins and cyclin-dependent kinases. Additionally, it can trigger apoptosis in various cancer cell lines through both intrinsic (mitochondrial) and extrinsic (death receptor) pathways. Biochanin A also inhibits aromatase activity, potentially reducing estrogen production, which may contribute to its effects against hormone-dependent cancers. In metabolic health, Biochanin A improves insulin sensitivity and glucose metabolism by enhancing insulin receptor signaling, GLUT4 translocation, and AMPK activation.

It also favorably modulates lipid metabolism by regulating genes involved in lipid synthesis and oxidation. For bone health, Biochanin A promotes osteoblast differentiation while inhibiting osteoclast activity, creating a positive balance in bone remodeling. It also enhances calcium absorption and retention, further supporting bone mineral density. Biochanin A is a potent inhibitor of drug transporters, particularly P-glycoprotein (P-gp), which can increase the bioavailability of certain medications.

It also inhibits fatty acid amide hydrolase (FAAH), the enzyme responsible for degrading endocannabinoids, potentially enhancing endocannabinoid signaling. Additionally, Biochanin A is a selective and reversible inhibitor of monoamine oxidase B (MAO-B), which may contribute to its neuroprotective effects in conditions like Parkinson’s disease. It also inhibits β-site APP-cleaving enzyme 1 (BACE1), a key enzyme in the pathogenesis of Alzheimer’s disease.

The optimal dosage of Biochanin A has not been definitively established through large-scale clinical trials in humans. Most research on Biochanin A has been conducted in vitro or in animal models, with limited human clinical studies. Biochanin A is typically consumed as part of red clover extracts or isoflavone supplements rather than as an isolated compound. Red clover supplements standardized to contain isoflavones (including Biochanin A, formononetin, genistein, and daidzein) typically provide 40-80 mg of total isoflavones per day, with Biochanin A comprising approximately 10-30 mg of this amount.

This dosage range is based on traditional use and preliminary clinical studies rather than comprehensive dose-finding trials. It’s important to note that Biochanin A is partially converted to genistein in the body, so its effects may be partially attributed to this metabolic conversion.

Biochanin A demonstrates poor oral bioavailability, which significantly limits its therapeutic potential. This low bioavailability is primarily attributed to its poor aqueous solubility, extensive first-pass metabolism, and efflux by intestinal transporters. When consumed orally, Biochanin A is absorbed in the small intestine, but the absorption rate is relatively low, with estimates ranging from 5-20% of the ingested dose reaching systemic circulation. After absorption, Biochanin A undergoes extensive phase I metabolism, primarily through O-demethylation by cytochrome P450 enzymes (particularly CYP1A1 and CYP1B1) to form genistein.

This conversion is significant because genistein possesses stronger estrogenic and tyrosine kinase inhibitory activities than Biochanin A itself. Additionally, Biochanin A undergoes phase II metabolism in the liver, where it is conjugated with glucuronic acid and sulfate. These conjugated forms constitute the majority (>95%) of circulating Biochanin A in plasma. The plasma elimination half-life of Biochanin A is relatively short, approximately 2-3 hours, which necessitates frequent dosing to maintain therapeutic levels.

Urinary excretion is the primary route of elimination for Biochanin A and its metabolites. Importantly, Biochanin A is a potent inhibitor of drug transporters, particularly P-glycoprotein (P-gp), which can increase the bioavailability of certain medications when co-administered. This property has led to interest in using Biochanin A as a bioavailability enhancer for poorly absorbed drugs, though this application requires careful consideration of potential drug interactions.

Nanostructured lipid carriers (NLCs) have shown significant improvement in Biochanin A bioavailability, with studies reporting up to 3.5-fold increase in oral bioavailability compared to unformulated Biochanin A, PEGylated nanostructured lipid carriers (PEG-NLCs) further enhance bioavailability by improving stability and circulation time, Mixed micelles containing Pluronic F127 and Plasdone S630 have demonstrated enhanced oral bioavailability of Biochanin A, Inclusion complexes with cyclodextrins, particularly (2-hydroxypropyl)-β-cyclodextrin, improve solubility and dissolution rate, Enteric-coated microparticles protect Biochanin A from degradation in the stomach and enhance intestinal absorption, Liposomal formulations enhance cellular uptake and improve bioavailability, Taking Biochanin A supplements with meals containing some fat may enhance absorption, Co-administration with piperine (black pepper extract) may inhibit glucuronidation and increase bioavailability, Self-emulsifying drug delivery systems (SEDDS) improve solubility and enhance lymphatic transport, Solid lipid nanoparticles (SLNs) provide controlled release and improved bioavailability

Due to the limited clinical research on Biochanin A as an isolated compound, specific timing recommendations are not well-established. However, based on its pharmacokinetic properties and the available research on isoflavone supplements containing Biochanin A, the following general recommendations can be made: For optimal absorption, Biochanin A supplements should be taken with meals, particularly those containing some fat, as this may enhance dissolution and absorption. Given its relatively short half-life (2-3 hours), dividing the daily dose into two administrations (morning and evening) may provide more consistent plasma levels. For menopausal symptom relief, consistent daily consumption is more important than specific timing, as the beneficial effects typically develop over weeks of regular use.

For bone health support, taking Biochanin A together with calcium and vitamin D supplements may be beneficial, as these nutrients work synergistically. For cardiovascular benefits, taking Biochanin A with meals may help reduce postprandial oxidative stress and lipid peroxidation. When using Biochanin A as part of red clover or isoflavone supplements for menopausal symptoms, taking the last dose of the day approximately 1-2 hours before bedtime may be helpful for night sweats. If using Biochanin A supplements alongside medications, be aware of potential interactions due to its inhibitory effects on drug transporters like P-glycoprotein.

Consult a healthcare provider for specific guidance on timing to minimize interactions.

• Gastrointestinal discomfort (mild bloating, gas, nausea) – most common, typically mild
• Headache – uncommon
• Skin rash – rare
• Menstrual cycle changes in premenopausal women – uncommon
• Insomnia or sleep disturbances – rare

• Current or history of estrogen-receptor positive breast cancer (controversial, consult healthcare provider)
• Current or history of endometrial cancer
• Undiagnosed abnormal uterine bleeding
• Active or history of thromboembolic disorders
• Known allergy or hypersensitivity to red clover or soy products
• Pregnancy and lactation (due to lack of safety data)
• Concurrent use of anticoagulant medications (potential interaction)
• Hormone-sensitive conditions that might be affected by phytoestrogens

• Tamoxifen and other selective estrogen receptor modulators (SERMs) – potential interference with therapeutic effects
• Aromatase inhibitors – potential interference with therapeutic effects
• Anticoagulants (warfarin, heparin) – Biochanin A may enhance anticoagulant effects, increasing bleeding risk
• P-glycoprotein substrates (digoxin, cyclosporine, certain chemotherapeutics) – Biochanin A inhibits P-gp, potentially increasing drug levels
• CYP450 substrates – Biochanin A inhibits various CYP enzymes, potentially affecting drug metabolism
• Estrogen-containing medications (including hormonal contraceptives) – additive estrogenic effects possible
• Antidiabetic medications – may enhance hypoglycemic effects
• Thyroid medications – theoretical interaction, though less likely than with other isoflavones

No official upper limit has been established by regulatory authorities specifically for Biochanin A. Clinical studies using red clover extracts have typically used preparations providing approximately 10-30 mg of Biochanin A daily (as part of 40-80 mg total isoflavones) without serious adverse effects in most populations. For general safety, most experts recommend not exceeding the amounts found in standard red clover supplements (approximately 30 mg/day of Biochanin A) for long-term use without medical supervision. Higher doses may be appropriate for specific therapeutic purposes under healthcare provider guidance, but safety data for high-dose, long-term use is limited.

The safety of isolated Biochanin A supplements at doses exceeding those found in traditional red clover extracts has not been thoroughly evaluated in large-scale human studies. Individuals with specific health conditions, particularly hormone-sensitive conditions, should consult healthcare providers before using supplements containing Biochanin A. Due to its inhibitory effects on drug transporters and metabolizing enzymes, Biochanin A may interact with various medications, potentially altering their efficacy or safety profiles. This is particularly important when considering higher doses.

In the United States, Biochanin A as a component of red clover extracts is regulated as a dietary supplement under the Dietary Supplement Health and Education Act (DSHEA) of 1994. As a dietary supplement ingredient, Biochanin A is not subject to the same pre-market approval process as pharmaceuticals. Manufacturers are responsible for ensuring their products are safe before marketing and that product labels are truthful and not misleading. The FDA does not authorize specific health claims for Biochanin A or red clover supplements.

Any claims made must be limited to general structure/function claims rather than disease treatment claims. Red clover extracts containing Biochanin A have not received a qualified health claim from the FDA for any specific health condition. The FDA has not established a specific upper limit for Biochanin A consumption. Isolated Biochanin A supplements (as opposed to red clover extracts) are less common in the US market, and their regulatory status would depend on whether they meet the definition of a dietary ingredient under DSHEA.

If Biochanin A were to be developed as a pharmaceutical agent for specific therapeutic applications, it would require formal FDA approval through the standard drug approval process, including clinical trials demonstrating safety and efficacy.

Eu: In the European Union, Biochanin A as a component of red clover extracts is regulated as a food supplement under the Food Supplements Directive (2002/46/EC). The European Food Safety Authority (EFSA) has evaluated several health claims for isoflavones, including those found in red clover, and has generally not approved specific claims due to insufficient evidence according to their standards. Isolated Biochanin A would likely be considered a novel food ingredient under Regulation (EU) 2015/2283 if marketed separately from traditional botanical extracts, requiring safety assessment before marketing. The EU has not established a specific upper limit for Biochanin A consumption. Some EU member states have implemented additional national regulations or recommendations regarding isoflavone supplements, which would include red clover extracts containing Biochanin A.

Canada: Health Canada regulates red clover extracts containing Biochanin A as Natural Health Products (NHPs). Several red clover products have received Natural Product Numbers (NPNs), indicating they have been assessed for safety, efficacy, and quality. Health Canada has approved certain claims for red clover supplements, primarily related to menopausal symptom relief. These claims typically require specific standardization of isoflavone content, including Biochanin A. Isolated Biochanin A supplements would require separate approval as NHPs. Health Canada has not established a specific upper limit for Biochanin A consumption but generally recommends doses consistent with those used in approved clinical studies.

Australia: The Therapeutic Goods Administration (TGA) regulates red clover extracts containing Biochanin A as complementary medicines. Many red clover products are listed on the Australian Register of Therapeutic Goods (ARTG) as AUST L products, which are assessed for safety and quality but not efficacy. Some specific red clover products with substantial evidence may be registered as AUST R products, which undergo more rigorous assessment including efficacy evaluation. Isolated Biochanin A supplements would require separate listing or registration. The TGA has not established a specific upper limit for Biochanin A consumption but generally follows the safety assessments conducted by other major regulatory bodies.

Japan: In Japan, red clover extracts containing Biochanin A may be regulated as Foods with Health Claims, specifically as Foods with Functional Claims (FFC) if scientific evidence supports their benefits. Manufacturers must notify the Consumer Affairs Agency before marketing such products. Isolated Biochanin A supplements are not commonly marketed in Japan. The Japanese Ministry of Health, Labour and Welfare has not established a specific upper limit for Biochanin A consumption.

Medium

Biochanin A is typically consumed as part of red clover extracts rather than as an isolated compound. Red clover supplements standardized to contain isoflavones (including Biochanin A, formononetin, genistein, and daidzein) typically cost between $0.30-$1.00 per day for an effective dose (40-80 mg of total isoflavones, containing approximately 10-25 mg of Biochanin A). Generic red clover extracts are generally the most affordable ($0.30-$0.60 per day), while specialized formulations with enhanced bioavailability, specific isoflavone profiles, or additional synergistic ingredients can cost $0.70-$1.20 per day. Isolated Biochanin A supplements are rare in the consumer market and are primarily available for research purposes at significantly higher costs.

Advanced delivery systems designed to enhance Biochanin A bioavailability, such as nanostructured lipid carriers or liposomal formulations, would likely command premium prices if commercially available.

The value proposition of Biochanin A varies significantly depending on the specific health application and individual factors. For menopausal symptom relief, red clover extracts containing Biochanin A offer moderate value, with clinical studies showing mixed results. Some women experience significant benefits, while others report minimal effects. Compared to hormone replacement therapy, red clover extracts are generally less effective for severe symptoms but have fewer potential side effects and lower cost.

For bone health support, the value proposition is moderate, as effects are typically modest and require long-term use. However, when compared to other bone health supplements, red clover extracts may offer unique benefits through their complex isoflavone profile. For cardiovascular health, the value proposition is less clear, as benefits have been demonstrated primarily in short-term studies with limited long-term outcome data. For neuroprotective effects, which have been demonstrated primarily in preclinical studies, the value proposition remains theoretical until more human clinical data becomes available.

The poor bioavailability of Biochanin A significantly limits its value in standard supplement formulations. Advanced delivery systems that enhance bioavailability could potentially improve the cost-effectiveness of Biochanin A supplementation, but these are not widely available in consumer products. When comparing cost-efficiency across different sources, standard red clover extracts generally provide the highest Biochanin A content per dollar spent. However, the overall value depends on the specific health outcome targeted and individual response.

For most consumers, a balanced approach may offer the best value: incorporating Biochanin A-containing foods into the diet while selectively using standardized supplements for specific health concerns. The long-term value of Biochanin A supplementation may improve as research advances and more targeted, bioavailable formulations become available.

Biochanin A as a pure compound has moderate stability, with a typical shelf life of 2-3 years when stored properly under controlled conditions. However, in supplement formulations, particularly red clover extracts, the shelf life may be shorter (1-2 years) due to interactions with other compounds and exposure to environmental factors. The methoxy group in Biochanin A provides slightly better stability compared to its demethylated counterpart genistein, particularly in terms of oxidative stability. Liquid extracts typically have shorter shelf lives (12-18 months) compared to powdered or encapsulated forms.

Advanced formulations designed to enhance bioavailability, such as nanostructured lipid carriers or liposomal formulations, may have different stability profiles depending on the specific formulation technology used. Products with added antioxidants or stabilizers may maintain potency longer than those without such additives.

Store in a cool, dry place away from direct sunlight and heat sources. Optimal temperature range is 59-77°F (15-25°C). Keep in original container with lid tightly closed to protect from moisture, oxygen exposure, and light. Opaque, airtight containers are ideal for preserving potency.

Refrigeration is recommended after opening for liquid extracts or products without stabilizers. Avoid storing in bathroom medicine cabinets or kitchen areas where temperature and humidity fluctuate. For bulk powders, consider using desiccant packets to minimize moisture exposure. Freezing is not recommended for most formulations as freeze-thaw cycles can accelerate degradation.

If the product changes color significantly (becoming darker) or develops an unusual odor, it may indicate degradation and should be discarded. For advanced delivery systems like nanostructured lipid carriers or liposomal formulations, follow manufacturer-specific storage recommendations, as these may differ from conventional supplements.

Exposure to oxygen (oxidation is a primary degradation pathway for Biochanin A, particularly affecting the hydroxyl groups), Exposure to light, particularly UV light, which accelerates oxidation and structural changes, High temperatures (above 86°F/30°C) accelerate degradation, Alkaline conditions cause rapid degradation through ring opening reactions, High humidity, which can promote hydrolysis and microbial growth, Presence of metal ions, particularly iron and copper, which catalyze oxidation, Enzymatic degradation if moisture penetrates the product, Prolonged exposure to air after opening the container, Freeze-thaw cycles, which can disrupt the chemical structure, Microbial contamination, particularly in liquid formulations or products with high moisture content, Chemical interactions with other compounds in complex formulations, Demethylation reactions converting Biochanin A to genistein during storage

Synthesis Methods

Natural Sources

Quality Considerations

Biochanin A has been consumed by humans for millennia as a component of various plants, particularly red clover (Trifolium pratense), though it was not identified or isolated as a specific compound until relatively recently. Red clover has a long history of traditional use in various cultures for medicinal purposes. Native Americans used red clover for respiratory conditions, skin problems, and as a blood purifier. In traditional European herbal medicine, red clover was used for skin conditions like eczema and psoriasis, respiratory problems including whooping cough and bronchitis, and as a blood cleanser.

In traditional Chinese medicine, red clover was used to treat toxic abscesses and sores. While these traditional uses did not specifically target Biochanin A, this isoflavone is now known to be one of the primary bioactive compounds in red clover. The scientific understanding of Biochanin A began in the mid-20th century when isoflavones were first isolated and identified as phytochemicals. Biochanin A was specifically isolated and characterized in the 1950s, with its chemical structure determined through spectroscopic methods.

The estrogenic properties of isoflavones, including Biochanin A, weren’t fully recognized until the 1940s, when sheep grazing on clover-rich pastures in Western Australia experienced fertility problems. This phenomenon, known as ‘clover disease,’ led to the identification of isoflavones as the causative agents and the coining of the term ‘phytoestrogens.’ In the 1980s and 1990s, epidemiological studies began noting significantly lower rates of hormone-dependent cancers, cardiovascular disease, and menopausal symptoms in Asian populations consuming traditional soy-rich diets compared to Western populations. While soy contains primarily genistein and daidzein rather than Biochanin A, these observations sparked intense scientific interest in isoflavones as potential protective compounds. This research eventually expanded to include Biochanin A from red clover.

The 1990s saw the introduction of numerous red clover extract supplements to the market, particularly targeted at menopausal women seeking alternatives to hormone replacement therapy. These extracts were standardized to contain specific amounts of isoflavones, including Biochanin A. In the early 2000s, several clinical trials investigated the effects of red clover extracts on menopausal symptoms, bone health, and cardiovascular parameters, with mixed results. While these studies did not specifically isolate the effects of Biochanin A, they contributed to our understanding of the potential health benefits of isoflavone mixtures containing Biochanin A.

In recent years, research has increasingly focused on the specific properties and potential therapeutic applications of Biochanin A as an isolated compound, particularly its neuroprotective, anti-inflammatory, and metabolic effects. Advanced delivery systems have been developed to overcome Biochanin A’s poor bioavailability, potentially enhancing its therapeutic potential. Throughout this scientific evolution, red clover extracts containing Biochanin A have remained popular dietary supplements, particularly for women’s health. However, isolated Biochanin A supplements are not widely available commercially, and most human consumption of this compound continues to be through red clover extracts or through dietary sources.

No comprehensive meta-analyses specifically focused on Biochanin A are currently available. Most systematic reviews and meta-analyses have examined red clover extracts or isoflavones as a group, rather than Biochanin A specifically., Ghazanfarpour M, et al. Red clover for treatment of hot flashes and menopausal symptoms: A systematic review and meta-analysis. Journal of Obstetrics and Gynaecology. 2016;36(3):301-311. Found modest effects of red clover extracts (containing Biochanin A) on menopausal hot flashes, with significant heterogeneity among studies., Abdi F, et al. Effects of phytoestrogens on bone mineral density during the menopause transition: a systematic review of randomized, controlled trials. Climacteric. 2016;19(6):535-545. Included studies on red clover extracts (containing Biochanin A) and found some evidence for beneficial effects on bone mineral density in menopausal women.

Effects of Biochanin A on Cognitive Function in Postmenopausal Women (hypothetical – no actual registered trial), Biochanin A for Neuroprotection in Parkinson’s Disease (hypothetical – no actual registered trial), Biochanin A Supplementation for Metabolic Health in Type 2 Diabetes (hypothetical – no actual registered trial), Note: As of 2024, there are no major registered clinical trials specifically investigating isolated Biochanin A in humans. Most clinical research focuses on red clover extracts or isoflavone mixtures rather than Biochanin A specifically.