Eupatorin

• Anticancer
• Anti-inflammatory
• Antioxidant
• Diuretic

• Hepatoprotective
• Antimicrobial
• Antidiabetic
• Nephroprotective
• Cardiovascular protection

Eupatorin (5,3′-dihydroxy-6,7,4′-trimethoxyflavone) exerts its diverse biological effects through multiple molecular pathways, with some unique mechanisms that distinguish it from other flavonoids. One of eupatorin’s most distinctive and well-studied mechanisms is its selective activation in cancer cells through cytochrome P450 1 (CYP1) enzyme metabolism. Eupatorin itself has limited direct cytotoxicity against cancer cells. However, when metabolized by CYP1 enzymes (particularly CYP1A1 and CYP1B1), which are often overexpressed in tumor tissues but not in normal tissues, eupatorin is converted to its active metabolites through demethylation.

These metabolites, including luteolin and other hydroxylated derivatives, exhibit potent antiproliferative and cytotoxic effects. This CYP1-dependent activation represents a form of biocatalytic activation that provides a degree of tumor selectivity, as the active metabolites are preferentially generated in cancer cells with high CYP1 expression. In cancer cells, eupatorin and its metabolites induce cell cycle arrest primarily at the G2/M phase by modulating the expression and activity of cell cycle regulators. It decreases the expression of cyclin B1 and cyclin-dependent kinase 1 (CDK1), while increasing the expression of p21, a CDK inhibitor.

This disruption of cell cycle progression prevents cancer cells from dividing and proliferating. Eupatorin also induces apoptosis (programmed cell death) through both intrinsic (mitochondrial) and extrinsic (death receptor) pathways. It modulates the expression of Bcl-2 family proteins, decreasing anti-apoptotic proteins (Bcl-2, Bcl-xL) and increasing pro-apoptotic proteins (Bax, Bad). This leads to mitochondrial membrane permeabilization, cytochrome c release, and activation of caspase cascades.

Additionally, eupatorin activates the extrinsic apoptotic pathway by enhancing death receptor signaling and caspase-8 activation. As an anti-inflammatory agent, eupatorin inhibits the nuclear factor-kappa B (NF-κB) signaling pathway by preventing IκB kinase (IKK) activation and subsequent nuclear translocation of NF-κB, thereby reducing the expression of pro-inflammatory genes. It suppresses the production of inflammatory cytokines including tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6), while inhibiting cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) expression. Eupatorin also modulates the mitogen-activated protein kinase (MAPK) pathway, including p38, extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK), further contributing to its anti-inflammatory properties.

The antioxidant properties of eupatorin are mediated through both direct and indirect mechanisms. With its two hydroxyl groups, eupatorin can directly scavenge some reactive oxygen species (ROS), though its scavenging capacity is more limited than flavonoids with more hydroxyl groups. More significantly, eupatorin activates the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway, leading to increased expression of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and heme oxygenase-1 (HO-1). Eupatorin exhibits diuretic effects, which are attributed to its ability to inhibit Na+/K+/2Cl- cotransporter activity in the kidney, promoting sodium and water excretion.

This mechanism explains its traditional use in conditions requiring increased urine output and its presence in plants traditionally used for urinary tract conditions, such as Orthosiphon stamineus (Java tea). In the liver, eupatorin demonstrates hepatoprotective effects by reducing oxidative stress, inflammation, and lipid accumulation. It activates AMP-activated protein kinase (AMPK), which enhances fatty acid oxidation and reduces lipogenesis, potentially benefiting conditions like non-alcoholic fatty liver disease (NAFLD). Eupatorin also induces phase II detoxification enzymes through Nrf2 activation, enhancing the liver’s capacity to metabolize and eliminate toxins.

In metabolic regulation, eupatorin improves insulin sensitivity and glucose metabolism by activating AMPK and peroxisome proliferator-activated receptor gamma (PPAR-γ). It also inhibits α-glucosidase, an enzyme involved in carbohydrate digestion, potentially reducing postprandial glucose levels. The balanced hydroxyl/methoxy structure of eupatorin (two hydroxyl groups and three methoxy groups) contributes to its unique pharmacological profile. The methoxy groups enhance its lipophilicity and membrane permeability, while the hydroxyl groups maintain some direct antioxidant capacity.

This structural feature also influences its metabolism by CYP enzymes, particularly the tumor-selective activation by CYP1 enzymes, which is central to its anticancer mechanism.

Optimal dosage ranges for eupatorin in humans have not been well established through clinical trials. Most research has focused on eupatorin as a component of herbal extracts, particularly Orthosiphon stamineus (Java tea), rather than as an isolated compound. For Orthosiphon stamineus extracts, typical daily doses range from 2-9 grams of dried herb or 400-1200 mg of standardized extract containing 0.1-0.5% eupatorin. This would correspond to approximately 0.4-6 mg of eupatorin daily.

For isolated eupatorin, which is rarely available as a standalone supplement, estimated effective doses based on preclinical studies would range from 1-10 mg daily. It’s important to note that eupatorin’s bioactivation by CYP1 enzymes, particularly in cancer cells, suggests that its effective dose may be lower than many other flavonoids due to this targeted activation mechanism.

Eupatorin has moderate oral bioavailability, estimated at approximately 15-25% in animal studies. This is higher than many other flavonoids due to its balanced hydroxyl/methoxy structure, which provides a good compromise between water solubility and lipophilicity. The three methoxy groups (at positions 6, 7, and 4′) increase lipophilicity compared to more hydroxylated flavonoids, potentially enhancing passive diffusion across cell membranes. However, eupatorin’s bioavailability is still limited by several factors, including first-pass metabolism in the liver, efflux by P-glycoprotein transporters in the intestine, and phase II metabolism (primarily glucuronidation and sulfation).

Interestingly, eupatorin’s metabolism by cytochrome P450 enzymes, particularly CYP1A1 and CYP1B1, which are often overexpressed in tumor tissues, leads to the formation of active metabolites through demethylation. These metabolites, including luteolin and other hydroxylated derivatives, may have different bioavailability profiles and contribute to eupatorin’s overall biological effects. This metabolic activation is particularly relevant for eupatorin’s anticancer properties, as it provides a degree of tumor selectivity.

Nanoemulsion formulations – can increase bioavailability by 3-10 fold by improving solubility and enhancing intestinal permeability, Liposomal encapsulation – protects eupatorin from degradation and enhances cellular uptake, Self-emulsifying drug delivery systems (SEDDS) – improve dissolution and absorption in the gastrointestinal tract, Phospholipid complexes – enhance lipid solubility and membrane permeability, Microemulsions – provide a stable delivery system with enhanced solubility, Combination with piperine – inhibits P-glycoprotein efflux and intestinal metabolism, Cyclodextrin inclusion complexes – improve aqueous solubility while maintaining stability, Solid dispersion techniques – enhance dissolution rate and solubility, Co-administration with other flavonoids that may compete for metabolic enzymes, potentially extending eupatorin’s half-life, Nanoparticle formulations – improve stability and targeted delivery, particularly relevant for anticancer applications

Eupatorin is best absorbed when taken with meals containing some fat, which can enhance solubility and stimulate bile secretion, improving dissolution and absorption. The presence of other flavonoids may enhance eupatorin’s bioavailability through competitive inhibition of metabolic enzymes or transporters. For diuretic effects, morning dosing is recommended to avoid nighttime urination. For anti-inflammatory and antioxidant effects, timing is less critical than consistency of use, though divided doses throughout the day may maintain more consistent blood levels due to eupatorin’s relatively short half-life (approximately 3-5 hours in animal studies).

For metabolic support, taking eupatorin with meals may enhance its effects on postprandial glucose levels through α-glucosidase inhibition. Enhanced delivery formulations like nanoemulsions or liposomes may have different optimal timing recommendations based on their specific pharmacokinetic profiles, but generally follow the same principles of taking with food for optimal absorption. For individuals interested in eupatorin’s anticancer potential, it’s worth noting that the CYP1-dependent activation mechanism may be influenced by various factors, including the expression and activity of CYP enzymes, which can be affected by diet, medications, and circadian rhythms. However, the clinical relevance of these factors for supplementation is not well established, and eupatorin should not be used as a standalone cancer treatment.

• Gastrointestinal discomfort (mild to moderate)
• Nausea (uncommon)
• Diarrhea (uncommon)
• Headache (rare)
• Dizziness (rare)
• Allergic reactions (rare)
• Increased urination (expected due to diuretic effect)
• Electrolyte imbalances with high doses or prolonged use (due to diuretic effect)
• Mild hypotension (uncommon)

• Pregnancy and breastfeeding (due to insufficient safety data)
• Scheduled surgery (discontinue 2 weeks before due to potential anticoagulant effects)
• Bleeding disorders (due to potential antiplatelet activity)
• Hormone-sensitive conditions (due to potential phytoestrogenic effects)
• Individuals taking medications metabolized by CYP1A1, CYP1B1, or other CYP enzymes (due to potential interactions)
• Individuals with severe liver or kidney disease (due to limited data on metabolism and excretion in these populations)
• Individuals with electrolyte imbalances (due to diuretic effects)
• Individuals with dehydration (due to diuretic effects)

• Diuretic medications (may enhance diuretic effects, potentially leading to excessive fluid loss and electrolyte imbalances)
• Antihypertensive medications (may enhance blood pressure-lowering effects)
• Cytochrome P450 substrates (eupatorin may affect the metabolism of drugs that are substrates for CYP enzymes, particularly CYP1A1 and CYP1B1)
• P-glycoprotein substrates (may alter drug transport and absorption)
• Anticoagulant and antiplatelet medications (may enhance bleeding risk due to potential antiplatelet effects)
• Lithium (diuretic effects may increase lithium concentrations, potentially leading to toxicity)
• Digoxin (electrolyte imbalances from diuretic effects may increase risk of digoxin toxicity)
• Antidiabetic medications (may enhance blood glucose-lowering effects)
• Chemotherapeutic agents (potential interactions due to CYP1-mediated metabolism, which could either enhance or reduce efficacy depending on the specific drug)

Due to limited human clinical data on isolated eupatorin, a definitive upper limit has not been established. Based on safety data for Orthosiphon stamineus extracts (which contain eupatorin) and animal toxicity studies, doses up to 10 mg of eupatorin daily or 1200 mg of standardized Orthosiphon stamineus extract daily appear to be well-tolerated in most individuals. For general supplementation, doses exceeding these levels are not recommended without medical supervision due to potential drug interactions, diuretic effects, and limited long-term safety data at higher doses. It’s important to note that eupatorin’s metabolism by CYP enzymes suggests potential for drug interactions that should be considered when determining appropriate dosing.

Additionally, the diuretic effects of eupatorin may lead to electrolyte imbalances with high doses or prolonged use, particularly in susceptible individuals.

Eupatorin itself is not approved as a drug by the FDA and is not commonly available as an isolated supplement. Plant extracts containing eupatorin, such as Orthosiphon stamineus (Java tea) extracts, are regulated as dietary supplements under the Dietary Supplement Health and Education Act (DSHEA) of 1994. Manufacturers cannot make specific disease treatment claims but may make general structure/function claims with appropriate disclaimers. The FDA has not evaluated the safety or efficacy of eupatorin specifically.

Orthosiphon stamineus is generally recognized as safe (GRAS) when used in traditional amounts as a tea or supplement.

Eu: In the European Union, eupatorin is not approved as a medicinal product. Orthosiphon stamineus extracts containing eupatorin are regulated as traditional herbal medicinal products under Directive 2004/24/EC in several EU countries, allowing them to be sold with specific health claims related to traditional use for increasing the amount of urine to achieve flushing of the urinary tract and for minor urinary complaints. In other EU countries, these extracts may be sold as food supplements, subject to the general food safety regulations. The European Medicines Agency (EMA) has published a community herbal monograph on Orthosiphon stamineus, recognizing its traditional medicinal use.

Germany: In Germany, Orthosiphon stamineus is approved by Commission E (the German regulatory authority for herbs) for irrigation therapy in inflammatory conditions of the lower urinary tract and prevention of kidney gravel. It is available as a traditional herbal medicinal product.

Malaysia: In Malaysia, where Orthosiphon stamineus is native and widely used, it is registered with the National Pharmaceutical Regulatory Agency (NPRA) as a traditional medicine. Several products containing Orthosiphon stamineus extracts are approved for traditional uses including diuresis, kidney and bladder inflammation, and gout.

Indonesia: In Indonesia, Orthosiphon stamineus is officially listed in the Indonesian Herbal Pharmacopoeia and is approved as a traditional medicine (jamu) for diuretic purposes, kidney stones, and rheumatism.

Australia: The Therapeutic Goods Administration (TGA) regulates Orthosiphon stamineus extracts as complementary medicines. Several products containing these extracts are listed on the Australian Register of Therapeutic Goods (ARTG). Traditional use claims are permitted with appropriate evidence of traditional use. Eupatorin as an isolated compound is not specifically regulated.

Japan: Orthosiphon stamineus is recognized as a medicinal plant in Japan and is included in some Kampo formulations. Eupatorin as an isolated compound is not specifically regulated for therapeutic use.

Medium (as part of herbal extracts) / High (as isolated compound)

Isolated eupatorin is rarely available commercially for supplementation and is primarily sold as a research chemical at prices ranging from $300-$800 per 10-25 mg, making

it prohibitively expensive for regular supplementation. Standardized Orthosiphon stamineus extracts containing eupatorin along with other flavonoids typically cost $0.30-$1.50 per day for basic extracts and $1.50-$4.00 per day for premium, highly standardized formulations. Orthosiphon stamineus tea, the most traditional form of consumption, is the most cost-effective option, typically costing $0.20-$0.80 per day.

The cost-effectiveness of eupatorin must be evaluated in the context of herbal extracts containing it, as isolated eupatorin is not practically available for regular supplementation due to its high cost and limited commercial availability. For diuretic effects and urinary tract health, Orthosiphon stamineus tea or basic extracts offer good value due to their relatively low cost and established traditional use. The diuretic effects are typically observed within a few days of consistent use, providing a reasonable return on investment for this specific application. For anti-inflammatory and antioxidant benefits, Orthosiphon stamineus extracts offer moderate value compared to other botanical anti-inflammatories.

While not as potent as some specialized anti-inflammatory supplements, they provide a broader spectrum of benefits at a lower cost. For metabolic support, particularly for blood glucose management, the value proposition is less clear. While preclinical studies suggest potential benefits, the clinical evidence is limited, and there may be more cost-effective options specifically for this application. For anticancer applications, the unique CYP1-dependent activation mechanism of eupatorin is scientifically interesting but not yet translated into practical therapeutic applications.

The current state of research does not support the use of eupatorin or Orthosiphon extracts as standalone cancer treatments, regardless of cost considerations. When comparing the cost-effectiveness of Orthosiphon extracts containing eupatorin to other supplements with similar indications: For diuretic effects, Orthosiphon extracts are comparable in cost to dandelion and juniper berry but may offer more comprehensive benefits due to the presence of multiple active compounds including eupatorin, sinensetin, and rosmarinic acid. For urinary tract health, Orthosiphon extracts are generally more cost-effective than cranberry extracts while providing complementary benefits. For general antioxidant and anti-inflammatory benefits, Orthosiphon extracts are moderately priced compared to alternatives, offering good but not exceptional value.

The most cost-effective way to consume eupatorin is through traditional Orthosiphon stamineus tea, which can be prepared from dried leaves at a fraction of the cost of processed extracts. However, the concentration of eupatorin and other active compounds may be lower and less consistent in tea preparations compared to standardized extracts.

Pure eupatorin is moderately stable, with a typical shelf life of 2-3 years when properly stored. The balanced hydroxyl/methoxy structure (two hydroxyl groups and three methoxy groups) provides better stability compared to more hydroxylated flavonoids. Standardized herbal extracts containing eupatorin, such as Orthosiphon stamineus extracts, typically have a shelf life of 1-2 years from the date of manufacture. Tea preparations have a much shorter shelf life, with optimal potency maintained for 6-12 months when properly stored.

Enhanced delivery formulations such as nanoemulsions or liposomes generally have shorter shelf lives of 1-2 years, depending on the specific formulation and preservative system.

Store in a cool, dry place away from direct sunlight in airtight, opaque containers. Refrigeration is recommended for liquid formulations and can extend shelf life of extracts containing eupatorin. Protect from moisture, heat, oxygen, and light exposure, which can accelerate degradation. For research-grade pure eupatorin, storage under inert gas (nitrogen or argon) at -20°C is recommended for maximum stability.

For dried herb material (e.g., Orthosiphon stamineus leaves), store in airtight containers away from light and moisture to preserve the eupatorin content. Tea preparations should be consumed fresh, as eupatorin and other active compounds may degrade over time in aqueous solutions. The addition of antioxidants such as vitamin E or ascorbic acid to formulations can help prevent oxidation and extend shelf life. Enhanced delivery formulations may have specific storage requirements provided by the manufacturer, which should be followed carefully to maintain stability and potency.

Avoid repeated freeze-thaw cycles, particularly for liquid formulations, as this can destabilize the product.

Exposure to UV light and sunlight – causes photodegradation, though the methoxy groups provide some protection compared to more hydroxylated flavonoids, High temperatures (above 30°C) – accelerates decomposition, Moisture – can promote hydrolysis and microbial growth, particularly in liquid formulations, Oxygen exposure – leads to oxidation, particularly of the hydroxyl groups, pH extremes – eupatorin is most stable at slightly acidic to neutral pH (5-7), Metal ions (particularly iron and copper) – can catalyze oxidation reactions, Enzymatic activity – may occur in improperly processed plant extracts, Incompatible excipients in formulations – certain preservatives or other ingredients may interact negatively with eupatorin, Repeated freeze-thaw cycles – can destabilize enhanced delivery formulations such as nanoemulsions or liposomes

Synthesis Methods

Natural Sources

Quality Considerations

Eupatorin itself was not identified or isolated until the modern era, but it is a constituent of several plants that have been used in traditional medicine systems for centuries. While the specific contribution of eupatorin to the traditional uses of these plants was unknown to ancient practitioners, it is now recognized as one of the bioactive compounds in these historically important medicinal materials. Eupatorin is primarily found in Orthosiphon stamineus (also known as Java tea, Cat’s whiskers, or Misai Kucing) and Eupatorium cannabinum (Hemp agrimony), both of which have rich histories in traditional medicine. Orthosiphon stamineus has been used in Southeast Asian traditional medicine, particularly in Indonesia, Malaysia, Thailand, and Vietnam, for hundreds of years.

In traditional Indonesian and Malaysian medicine (Jamu), it was classified as a plant that promotes diuresis and detoxification. The leaves were traditionally prepared as a tea and used to treat various conditions including kidney and bladder inflammation, kidney stones, urinary tract infections, diabetes, hypertension, rheumatism, and gout. The first documented medicinal use of Orthosiphon stamineus appears in traditional Javanese medical texts from the 18th century, where it was recommended for urinary disorders and ‘cleansing the kidneys.’ Dutch colonists in Indonesia became aware of its medicinal properties and introduced it to Europe in the late 19th century, where it gained popularity as ‘Java tea’ for treating kidney and bladder ailments. In Vietnamese traditional medicine, Orthosiphon stamineus (known as ‘Râu Mèo’ or cat’s whiskers) was used for similar purposes, with additional applications for treating edema and inflammatory conditions.

In Thai traditional medicine, it was known as ‘Yaa Nuat Maeo’ and was used for diuretic, antidiabetic, and anti-inflammatory purposes. Eupatorium cannabinum (Hemp agrimony) has been used in European traditional medicine since ancient times. The Greek physician Dioscorides mentioned it in his De Materia Medica in the 1st century CE, noting its use for liver ailments. In medieval European herbalism, it was used as a ‘blood purifier,’ diuretic, and liver tonic.

The plant was also used to treat fever, particularly malaria, earning it the name ‘fever plant’ in some regions. Native American tribes also used related Eupatorium species for similar purposes. The name ‘eupatorin’ is derived from Eupatorium, which itself was named after Mithridates Eupator, king of Pontus in the 1st century BCE, who was said to have discovered the medicinal properties of a plant in this genus. Modern scientific interest in eupatorin began in the mid-20th century as part of the scientific investigation into the active components of these traditional medicinal plants.

Its structure was elucidated as 5,3′-dihydroxy-6,7,4′-trimethoxyflavone, identifying it as a polymethoxylated flavone with a unique pattern of hydroxylation and methoxylation. In recent decades, research has revealed eupatorin’s potential anticancer properties, particularly its selective activation by CYP1 enzymes that are often overexpressed in tumor tissues. This mechanism was unknown to traditional practitioners but may explain some of the observed benefits of plants containing eupatorin in traditional medicine. Today, Orthosiphon stamineus extracts containing eupatorin are commercially available as dietary supplements and herbal teas, primarily marketed for kidney and urinary tract health, based on both traditional use and modern scientific research.

No meta-analyses specifically on eupatorin are currently available; most analyses focus on Orthosiphon stamineus extracts rather than isolated eupatorin.

Limited ongoing trials specifically investigating eupatorin; most research focuses on Orthosiphon stamineus extracts or other plant sources containing eupatorin, Several preclinical studies investigating eupatorin’s potential in cancer therapy, particularly focusing on its CYP1-dependent activation mechanism, Research on novel delivery systems to enhance eupatorin’s bioavailability and targeted delivery, Investigations into eupatorin’s potential for metabolic disorders, including diabetes, Studies on the combination of eupatorin with conventional therapies for enhanced efficacy in various conditions