Ononin

• Anti-inflammatory
• Antioxidant
• Cardiovascular protection
• Anticancer potential

• Neuroprotection
• Bone health
• Immunomodulation
• Metabolic regulation
• Antimicrobial

Ononin (formononetin-7-O-glucoside) exerts its diverse biological effects through multiple molecular pathways. As an O-glycosylated isoflavone, ononin possesses a unique structural feature where a glucose molecule is attached to the C-7 position of the formononetin backbone via an oxygen atom (O-glycosidic bond). This structure influences its pharmacokinetics, metabolism, and biological activities. In the body, ononin undergoes metabolism primarily through hydrolysis of the O-glycosidic bond by intestinal β-glucosidases, releasing the aglycone formononetin.

This conversion is significant because formononetin is the biologically active form responsible for most of ononin’s health effects. The rate and extent of this conversion vary among individuals based on their gut microbiome composition, diet, and other factors. Formononetin can be further demethylated to form daidzein, which can be metabolized by gut microbiota to produce equol in some individuals. This metabolic conversion is significant because equol has potent estrogenic and antioxidant activities.

However, the ability to produce equol varies among individuals based on their gut microbiome composition, with approximately 30-50% of adults being equol producers. As a phytoestrogen precursor, ononin (through its metabolites formononetin and potentially daidzein and equol) demonstrates weak estrogenic activity due to structural similarity to 17β-estradiol. Formononetin binds to estrogen receptors (ERs), with a higher affinity for ER-β compared to ER-α. This selective ER modulation contributes to ononin’s potential benefits for hormone-dependent conditions while potentially reducing risks associated with ER-α activation.

The estrogenic effects are context-dependent, showing estrogen-like effects in low-estrogen environments (such as postmenopausal women) and potentially anti-estrogenic effects in high-estrogen environments through competitive binding to ERs. Ononin demonstrates significant anti-inflammatory effects, primarily through its metabolite formononetin. It 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.

Ononin also modulates the mitogen-activated protein kinase (MAPK) signaling pathways, including p38 MAPK, extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK), further contributing to its anti-inflammatory properties. As an antioxidant, ononin (through its metabolites) scavenges reactive oxygen species (ROS) and free radicals. It also enhances endogenous antioxidant defenses by activating nuclear factor erythroid 2-related factor 2 (Nrf2), a transcription factor that regulates the expression of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and heme oxygenase-1 (HO-1). In cardiovascular health, ononin (through its metabolite formononetin) improves endothelial function by increasing nitric oxide (NO) production through activation of endothelial nitric oxide synthase (eNOS).

It also demonstrates vasodilatory effects by activating large-conductance calcium-activated potassium (BKCa) channels in vascular smooth muscle cells. Ononin inhibits platelet aggregation and thrombus formation, potentially reducing the risk of thrombotic events. Additionally, it improves lipid profiles by reducing total cholesterol, low-density lipoprotein (LDL) cholesterol, and triglycerides while increasing high-density lipoprotein (HDL) cholesterol. Ononin has demonstrated anticancer potential through multiple mechanisms, primarily mediated by its metabolite formononetin.

It inhibits cell proliferation by inducing cell cycle arrest, primarily at the G0/G1 or G2/M phases, through modulation of cyclins, cyclin-dependent kinases (CDKs), and CDK inhibitors. It induces apoptosis (programmed cell death) in various cancer cell lines through both intrinsic (mitochondrial) and extrinsic (death receptor) pathways. It upregulates pro-apoptotic proteins (Bax, Bad) and downregulates anti-apoptotic proteins (Bcl-2, Bcl-xL), leading to mitochondrial membrane permeabilization, cytochrome c release, and caspase activation. Ononin also inhibits angiogenesis (formation of new blood vessels) by downregulating vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α (HIF-1α), thereby limiting tumor growth and metastasis.

Additionally, it suppresses cancer cell migration and invasion by inhibiting matrix metalloproteinases (MMPs) and modulating epithelial-mesenchymal transition (EMT) markers. In the central nervous system, ononin exhibits neuroprotective effects through multiple mechanisms. It reduces oxidative stress and neuroinflammation, protects against excitotoxicity, and promotes neuronal survival. Ononin also enhances the expression of neurotrophic factors, including brain-derived neurotrophic factor (BDNF), and activates the PI3K/Akt/glycogen synthase kinase-3β (GSK-3β) pathway, promoting neuronal survival and synaptic plasticity.

For bone health, ononin (through its metabolites) inhibits osteoclast differentiation and activity while promoting osteoblast proliferation and differentiation, potentially leading to increased bone formation and reduced bone resorption. These effects are mediated through both ER-dependent and ER-independent pathways, including modulation of the receptor activator of nuclear factor kappa-B ligand (RANKL)/osteoprotegerin (OPG) system. Ononin demonstrates immunomodulatory effects by regulating the balance between pro-inflammatory and anti-inflammatory cytokines, modulating T cell differentiation, and enhancing natural killer (NK) cell activity. It also exhibits antimicrobial properties against various bacteria and fungi, potentially through disruption of cell membranes and inhibition of essential microbial enzymes.

In metabolic regulation, ononin (through its metabolites) improves insulin sensitivity and glucose metabolism through multiple mechanisms. It activates adenosine monophosphate-activated protein kinase (AMPK) in skeletal muscle and liver, leading to increased glucose uptake, enhanced glycolysis, and reduced gluconeogenesis. It also promotes the translocation of glucose transporter 4 (GLUT4) to the cell membrane in muscle and adipose tissue, further enhancing glucose uptake. The O-glycosidic bond in ononin makes it more susceptible to hydrolysis compared to C-glycosides, affecting its bioavailability and metabolism.

This structural feature results in significant first-pass metabolism, with most ononin being converted to formononetin before reaching systemic circulation. Therefore, many of ononin’s biological effects are likely mediated through its metabolite formononetin rather than the parent compound itself.

Optimal dosage ranges for ononin are not well-established due to limited clinical studies specifically evaluating ononin as a standalone supplement. Most research has been conducted on plant extracts containing ononin along with other bioactive compounds. Based on the available research and traditional use, the following dosage ranges can be considered: For standardized Ononis spinosa extract (typically containing 1-3% ononin), the common dosage range is 300-900 mg daily, corresponding to approximately 3-27 mg of ononin. For standardized red clover extract (typically containing 0.5-2% ononin), typical dosages range from 40-160 mg daily, corresponding to approximately 0.2-3.2 mg of ononin.

For standardized Astragalus extract (typically containing 0.1-0.5% ononin), typical dosages range from 500-1000 mg daily, corresponding to approximately 0.5-5 mg of ononin. Isolated ononin supplements are rare, but when available, typical dosages would range from 5-25 mg daily, based on its proportional content in effective herbal extracts. It’s important to note that ononin’s bioavailability and metabolism can vary significantly between individuals based on gut microbiome composition, particularly regarding its conversion to formononetin and potentially to daidzein and equol. For most health applications, starting with a lower dose and gradually increasing as needed and tolerated is recommended.

Divided doses (2-3 times daily) may be preferred due to the relatively short half-life of ononin’s metabolites, though specific pharmacokinetic data in humans is limited.

Ononin has relatively low oral bioavailability, estimated at approximately 5-15% based on limited animal studies, though comprehensive human pharmacokinetic data is lacking. As an O-glycosylated isoflavone, ononin’s bioavailability is significantly influenced by its metabolism in the gastrointestinal tract. Upon oral administration, ononin undergoes significant first-pass metabolism in the intestine. The O-glycosidic bond (where glucose is attached to the C-7 position of formononetin via an oxygen atom) is readily hydrolyzed by intestinal β-glucosidases, releasing the aglycone formononetin.

This conversion begins in the small intestine and continues in the large intestine, with most ononin being converted to formononetin before reaching systemic circulation. The bioavailability of intact ononin is estimated to be less than 5%, while the bioavailability of its metabolite formononetin is estimated to be around 10-20%. In the intestine, formononetin can be further demethylated by cytochrome P450 enzymes to form daidzein, which can be metabolized by gut microbiota to produce equol in some individuals. This metabolic conversion is significant because equol has potent estrogenic and antioxidant activities.

However, the ability to produce equol varies among individuals based on their gut microbiome composition, with approximately 30-50% of adults being equol producers. In the liver, formononetin undergoes phase II metabolism, primarily through glucuronidation and sulfation, forming conjugates that are more water-soluble and readily excreted. These conjugates may be less biologically active than free formononetin, though some evidence suggests they can be deconjugated in target tissues, releasing the active compound. The plasma half-life of formononetin (the primary active metabolite of ononin) is relatively short, estimated at approximately 4-8 hours based on animal studies, necessitating multiple daily doses for sustained therapeutic effects.

Formononetin demonstrates moderate distribution to various tissues, with some evidence suggesting preferential accumulation in hormone-responsive tissues such as breast, uterus, and prostate, likely due to its interaction with estrogen receptors in these tissues.

Liposomal formulations – can increase bioavailability by 2-4 fold by enhancing cellular uptake and protecting ononin from degradation, Nanoemulsion formulations – can increase bioavailability by 3-5 fold by improving solubility and enhancing intestinal permeability, Self-emulsifying drug delivery systems (SEDDS) – improve dissolution and absorption in the gastrointestinal tract, Phospholipid complexes – enhance lipid solubility and membrane permeability, Cyclodextrin inclusion complexes – improve aqueous solubility while maintaining stability, Solid dispersion techniques – enhance dissolution rate and solubility, Combination with piperine – inhibits P-glycoprotein efflux and intestinal metabolism, potentially increasing bioavailability by 30-60%, Microemulsions – provide a stable delivery system with enhanced solubility, Co-administration with fatty meals – can increase absorption by stimulating bile secretion and enhancing lymphatic transport, Combination with probiotics – certain probiotic strains may enhance the conversion of ononin to formononetin and potentially to daidzein and equol, increasing its biological activity in equol producers

Ononin 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 dietary fiber may reduce absorption, so supplements may be more effective than whole food sources for achieving specific therapeutic effects. Due to the relatively short half-life of ononin’s metabolite formononetin (estimated at 4-8 hours based on animal studies), divided doses (2-3 times daily) may be beneficial for maintaining consistent blood levels throughout the day, though specific human pharmacokinetic data is limited. For anti-inflammatory support, consistent daily dosing is recommended, with some evidence suggesting that taking ononin with meals may help reduce gastrointestinal inflammation.

For cardiovascular support, consistent daily dosing is recommended, with some evidence suggesting that morning dosing may be particularly beneficial for blood pressure regulation, though more research is needed. For anticancer support, consistent daily dosing is important to maintain therapeutic levels in target tissues. Some research suggests that timing may influence efficacy, with potential benefits to taking ononin during specific phases of cancer treatment, though this requires medical supervision. For bone health, consistent daily dosing is important, as these effects develop gradually over time with regular use.

For urinary tract support, some traditional sources recommend taking Ononis spinosa extracts (containing ononin) in the morning and early afternoon rather than evening to avoid nighttime urination due to its mild diuretic effects. Enhanced delivery formulations like liposomes or nanoemulsions 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. The timing of ononin supplementation relative to other medications should be considered, as it may interact with certain drugs, particularly those affecting hormone levels or those metabolized by the same enzymes. In general, separating ononin supplementation from other medications by at least 2 hours is recommended to minimize potential interactions.

• Gastrointestinal discomfort (mild to moderate, common)
• Nausea (uncommon)
• Headache (uncommon)
• Menstrual changes in women (uncommon, due to phytoestrogenic effects of metabolites)
• Breast tenderness (rare, due to phytoestrogenic effects of metabolites)
• Allergic reactions (rare, particularly in individuals with legume allergies)
• Mild dizziness (rare)
• Skin rash (rare)
• Mild insomnia (rare)
• Increased urination (common with Ononis spinosa extracts due to diuretic effects)

• Pregnancy and breastfeeding (due to phytoestrogenic effects of metabolites and insufficient safety data)
• Hormone-sensitive conditions including hormone-dependent cancers (breast, uterine, ovarian) due to phytoestrogenic effects of metabolites
• Individuals with legume allergies (particularly for red clover or Astragalus-derived ononin)
• Individuals with severe liver disease (due to potential effects on liver enzymes)
• Individuals scheduled for surgery (discontinue 2 weeks before due to potential effects on blood clotting)
• Children and adolescents (due to potential hormonal effects during development)
• Individuals with thyroid disorders (isoflavones may affect thyroid function in susceptible individuals)
• Individuals with estrogen receptor-positive breast cancer or a history of such cancer (due to potential estrogenic effects of metabolites)
• Individuals with endometriosis or uterine fibroids (conditions that may be estrogen-sensitive)
• Individuals with severe kidney disease (particularly for Ononis spinosa extracts due to diuretic effects)

• Hormone replacement therapy and hormonal contraceptives (may interfere with or enhance effects due to phytoestrogenic activity of metabolites)
• Tamoxifen and other selective estrogen receptor modulators (SERMs) (potential competitive binding to estrogen receptors by metabolites)
• Anticoagulant and antiplatelet medications (may enhance antiplatelet effects, potentially increasing bleeding risk)
• Diuretic medications (may enhance diuretic effects, particularly with Ononis spinosa extracts)
• Cytochrome P450 substrates (may affect the metabolism of drugs that are substrates for CYP1A2, CYP2C9, and CYP3A4)
• Thyroid medications (isoflavones may affect thyroid function in susceptible individuals)
• Antidiabetic medications (may enhance blood glucose-lowering effects)
• Drugs metabolized by UDP-glucuronosyltransferases (UGTs) (potential competition for these enzymes)
• Drugs with narrow therapeutic indices (warfarin, digoxin, etc.) require careful monitoring due to potential interactions
• Aromatase inhibitors (may counteract the effects of these drugs used in breast cancer treatment)

Based on preclinical studies and limited clinical data, the upper limit for ononin supplementation is generally considered to be 25-30 mg daily for most adults. For Ononis spinosa extracts (typically containing 1-3% ononin), upper limits of 900-1200 mg daily are generally considered safe. For red clover extracts (typically containing 0.5-2% ononin), upper limits of 160-200 mg daily have been used in clinical studies without significant adverse effects. For Astragalus extracts (typically containing 0.1-0.5% ononin), upper limits of 1000-1500 mg daily are generally considered safe.

Higher doses may increase the risk of hormonal effects and drug interactions, particularly in sensitive individuals. For general supplementation, doses exceeding these levels are not recommended without medical supervision. The safety profile of ononin is generally favorable at recommended doses, with most side effects being mild and transient. However, the phytoestrogenic properties of its metabolites and potential for drug interactions necessitate caution, particularly with long-term use or in vulnerable populations.

Individuals with hormone-sensitive conditions, thyroid disorders, or those taking medications with potential interactions should consult healthcare providers before use. The long-term safety of high-dose ononin supplementation has not been fully established, particularly regarding effects on hormone-sensitive tissues. Some regulatory authorities have expressed caution about long-term, high-dose isoflavone supplementation in certain populations, such as women with a history or family history of breast cancer. The potential for ononin’s metabolites to act as both estrogen agonists and antagonists, depending on the tissue, estrogen environment, and dose, adds complexity to safety considerations.

This dual activity may be beneficial in some contexts (such as bone health in postmenopausal women) but potentially harmful in others (such as in estrogen-sensitive cancers). It’s worth noting that most safety data for ononin comes from studies on plant extracts containing ononin along with other bioactive compounds, rather than isolated ononin. Therefore, the specific safety profile of isolated ononin may differ from that of the plant extracts.

In the United States, ononin is not approved by the FDA as a drug. Ononis spinosa, red clover, and Astragalus extracts containing ononin 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 ononin specifically.

Ononis spinosa, red clover, and Astragalus are generally recognized as safe (GRAS) when used in traditional amounts as herbs or supplements.

Eu: In the European Union, ononin is not approved as a medicinal product. Ononis spinosa, red clover, and Astragalus extracts are primarily regulated as food supplements under the Food Supplements Directive (2002/46/EC). However, Ononis spinosa is also included in the European Medicines Agency’s (EMA) ‘Community herbal monographs’ as a traditional herbal medicinal product for increasing the amount of urine to achieve flushing of the urinary tract as an adjuvant in minor urinary complaints. This allows products containing Ononis spinosa to be registered as traditional herbal medicinal products in EU member states, provided they meet the quality, safety, and traditional use requirements.

Uk: In the United Kingdom, Ononis spinosa, red clover, and Astragalus extracts are regulated as food supplements. Ononis spinosa can also be registered as a traditional herbal medicinal product for urinary tract conditions, similar to the EU regulation. They are not licensed as medicines for specific indications and cannot be marketed with medicinal claims unless registered as traditional herbal medicinal products.

Canada: Health Canada regulates Ononis spinosa, red clover, and Astragalus extracts as Natural Health Products (NHPs). Several products containing these extracts have been issued Natural Product Numbers (NPNs), allowing them to be sold with specific health claims, primarily related to traditional use in herbal medicine. Isolated ononin is not specifically approved as a standalone ingredient.

Australia: The Therapeutic Goods Administration (TGA) regulates Ononis spinosa, red clover, and Astragalus 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. Ononin as an isolated compound is not specifically regulated.

China: In China, Astragalus (Huangqi) is officially listed in the Chinese Pharmacopoeia as a traditional Chinese medicine and is approved for various indications including strengthening immunity, promoting tissue regeneration, and increasing vitality. Ononis spinosa and red clover are not as commonly used in traditional Chinese medicine but may be included in some formulations. Ononin as an isolated compound is primarily used in research rather than as an approved therapeutic agent.

Japan: In Japan, Astragalus is recognized as a traditional herbal medicine ingredient and is included in several approved Kampo formulations. Ononis spinosa and red clover extracts are available as dietary supplements but are not as commonly used as in Western countries. Ononin as an isolated compound is not specifically approved as a pharmaceutical but is available as a component of various dietary supplements.

Korea: In South Korea, Astragalus is recognized as a traditional herbal medicine and is included in the Korean Pharmacopoeia. Ononis spinosa and red clover extracts are available as dietary supplements. Ononin as an isolated compound is primarily used in research rather than as an approved therapeutic agent.

Medium to High

Isolated ononin supplements are rare and typically expensive when available, costing $2.00-$4.00 per day for effective doses (5-25 mg daily). Standardized Ononis spinosa extracts (containing ononin along with other bioactive compounds) typically cost $0.75-$2.00 per day for basic extracts (300-900 mg daily, corresponding to approximately 3-27 mg of ononin) and $2.00-$3.50 per day for premium, highly standardized formulations. Standardized red clover extracts (containing ononin along with other isoflavones) typically cost $0.30-$1.00 per day for basic extracts (40-160 mg daily, corresponding to approximately 0.2-3.2 mg of ononin) and $1.00-$2.00 per day for premium formulations. Standardized Astragalus extracts (containing ononin along with other bioactive compounds) typically cost $0.50-$1.50 per day for basic extracts (500-1000 mg daily, corresponding to approximately 0.5-5 mg of ononin) and $1.50-$3.00 per day for premium formulations.

Enhanced delivery formulations such as liposomes or nanoemulsions typically cost $3.00-$6.00 per day, though they may offer better bioavailability and potentially superior therapeutic outcomes.

For anti-inflammatory support, ononin offers moderate value. Preclinical studies have demonstrated significant anti-inflammatory effects through inhibition of the NF-κB pathway and modulation of inflammatory cytokines. While clinical evidence in humans is limited, the mechanisms of action are well-established, and the potential benefits align with traditional uses of plants containing ononin, particularly Ononis spinosa and Astragalus. When compared to other anti-inflammatory supplements, ononin (particularly as part of Ononis spinosa or Astragalus extracts) is moderately priced and offers a comprehensive approach to inflammation.

For cardiovascular support, ononin offers moderate value. Preclinical studies have demonstrated significant cardiovascular benefits, including antioxidant effects, improved endothelial function, and protection against oxidative stress-induced cardiomyocyte damage. While clinical evidence in humans is limited, the mechanisms of action are well-established, and the potential benefits align with traditional uses of plants containing ononin, particularly Astragalus. When compared to other cardiovascular supplements, ononin is relatively expensive but offers a unique mechanism of action through its specific isoflavone glycoside structure.

For anticancer support as a complementary approach, ononin offers moderate value. Preclinical studies have demonstrated anticancer effects through multiple mechanisms, including induction of apoptosis, cell cycle arrest, and inhibition of migration and invasion. However, clinical evidence in humans is lacking, and ononin should only be considered as a complementary approach alongside conventional cancer treatments, not as a standalone treatment. When compared to other natural compounds with anticancer potential, ononin is relatively expensive but offers a unique mechanism of action through its specific isoflavone glycoside structure.

For bone health, ononin offers moderate value. Preclinical studies have demonstrated bone-protective effects through inhibition of osteoclastogenesis and promotion of osteoblast activity. While clinical evidence in humans is limited, the mechanisms of action are well-established, and the potential benefits align with the known effects of isoflavones on bone metabolism. When compared to other bone health supplements, ononin is relatively expensive but offers a complementary approach that may be particularly beneficial when combined with calcium and vitamin D.

For urinary tract support, ononin (particularly as part of Ononis spinosa extracts) offers good value. Traditional use of Ononis spinosa for urinary tract conditions is well-documented, and some clinical evidence supports its mild diuretic effects. When compared to other urinary tract supplements, Ononis spinosa extracts are moderately priced and offer a comprehensive approach that aligns with traditional use. When comparing the cost-effectiveness of different sources of ononin: Standardized Ononis spinosa extracts offer a good balance of cost and standardized dosing for urinary tract conditions and general anti-inflammatory support.

They typically contain ononin alongside other bioactive compounds, providing a comprehensive approach that aligns with traditional use. Standardized red clover extracts are less expensive than Ononis spinosa extracts and provide ononin alongside other isoflavones like formononetin and biochanin A, which may offer synergistic effects for bone health and menopausal symptoms. Standardized Astragalus extracts are moderately priced and provide ononin alongside other bioactive compounds like astragalosides and polysaccharides, which may offer synergistic effects for immune support and cardiovascular health. Enhanced delivery formulations such as liposomes or nanoemulsions offer better bioavailability and potentially superior therapeutic outcomes, which may justify their higher cost for specific health conditions.

However, for general health maintenance, standard formulations are likely more cost-effective. Individual variation in ononin metabolism significantly affects the value proposition of ononin supplementation. Factors such as gut microbiome composition, diet, and genetic factors can influence the conversion of ononin to formononetin and potentially to daidzein and equol, leading to variable responses among individuals.

Pure ononin has moderate stability, with a typical shelf life of 1-2 years when properly stored. The O-glycosidic bond (where glucose is attached to the C-7 position of formononetin via an oxygen atom) is relatively stable under normal storage conditions but can be hydrolyzed under acidic conditions or in the presence of specific enzymes like β-glucosidases. Standardized plant extracts containing ononin (such as Ononis spinosa, red clover, or Astragalus extracts) typically have a shelf life of 1-2 years from the date of manufacture when properly stored. Dried plant material (Ononis spinosa root, red clover flowers, Astragalus root) properly stored can maintain acceptable ononin content for 1-2 years.

Traditional decoctions and liquid extracts have a much shorter shelf life, with optimal potency maintained for only a few days under refrigeration. Enhanced delivery formulations such as liposomes or nanoemulsions 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 ononin. Protect from moisture, heat, oxygen, and light exposure, which can accelerate degradation. For research-grade pure ononin, storage under inert gas (nitrogen or argon) at -20°C is recommended for maximum stability.

For dried plant material (Ononis spinosa root, red clover flowers, Astragalus root), store in airtight containers away from light and moisture to preserve the ononin content. 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.

For traditional decoctions, prepare fresh and consume within 24-48 hours, storing any remainder in the refrigerator.

Exposure to UV light and sunlight – causes photodegradation of the isoflavone structure, High temperatures (above 30°C) – accelerates decomposition and hydrolysis of the O-glycosidic bond, Moisture – promotes hydrolysis of the O-glycosidic bond and microbial growth, particularly in liquid formulations, Oxygen exposure – leads to oxidation, particularly affecting the hydroxyl group at the C-7 position, pH extremes – ononin is most stable at slightly acidic to neutral pH (5-7), with increased hydrolysis in strongly acidic conditions and increased degradation in alkaline conditions, Metal ions (particularly iron and copper) – can catalyze oxidation reactions, Enzymatic activity – β-glucosidases can hydrolyze the O-glycosidic bond, converting ononin to formononetin, Microbial contamination – particularly relevant for liquid formulations, can lead to degradation of active compounds through enzymatic hydrolysis, Incompatible excipients in formulations – certain preservatives or other ingredients may interact negatively with ononin, Repeated freeze-thaw cycles – can destabilize enhanced delivery formulations such as liposomes or nanoemulsions

Synthesis Methods

Natural Sources

Quality Considerations

Ononin itself was not identified or isolated until the modern era, but it is a bioactive constituent of several plants that have been used in traditional medicine systems for thousands of years. While the specific contribution of ononin to the traditional uses of these plants was unknown to ancient practitioners, it is now recognized as one of the compounds responsible for many of their medicinal properties. Ononis spinosa (Spiny restharrow) has been used in European traditional medicine since ancient times. It was mentioned by Dioscorides in his ‘De Materia Medica’ in the 1st century CE, where it was recommended primarily for its diuretic properties.

The plant’s Latin name ‘Ononis’ is derived from the Greek word ‘onos’ (donkey), as donkeys were said to eat the plant despite its spiny nature. The species name ‘spinosa’ refers to the plant’s thorny stems. In traditional European herbal medicine, Ononis spinosa root was primarily used for urinary tract conditions, including kidney stones, urinary tract infections, and edema. It was valued for its diuretic and anti-inflammatory properties, which align with modern research on ononin’s effects.

The root was typically prepared as a decoction or tincture. In addition to its urinary applications, Ononis spinosa was traditionally used for rheumatic complaints, gout, and skin conditions. These uses also align with modern research on ononin’s anti-inflammatory and antioxidant properties. In the Middle Ages, Ononis spinosa was included in various herbal compendiums and was recommended by influential herbalists such as Hildegard von Bingen (12th century) and Nicholas Culpeper (17th century).

Culpeper’s ‘Complete Herbal’ described restharrow as useful for ‘stone and gravel’ (kidney stones) and for ‘obstructions of the liver and spleen.’ In traditional German medicine, Ononis spinosa was one of the ingredients in ‘Species diureticae’ (diuretic species), a mixture of herbs used to promote urine flow and treat urinary tract conditions. This formulation was included in various pharmacopoeias until the early 20th century. Red clover (Trifolium pratense) has a rich history of use in European folk medicine dating back centuries. Traditional herbalists used red clover for various conditions, including respiratory ailments (such as whooping cough, bronchitis, and asthma), skin disorders (including eczema and psoriasis), and as a blood purifier.

It was also traditionally used for women’s health issues, though not specifically for menopausal symptoms as it is often used today. In the 19th century, red clover became part of various herbal formulations for treating cancer and other chronic diseases, most notably as an ingredient in the Hoxsey formula, a controversial alternative cancer treatment. Native American tribes, including the Cherokee, Iroquois, and Ojibwa, also used red clover for respiratory conditions, fever, and as a blood cleanser. Astragalus membranaceus (Huangqi) has an extensive documented history in traditional Chinese medicine (TCM), dating back over 2,000 years.

It was first described in the ‘Shennong Bencao Jing’ (Divine Farmer’s Classic of Materia Medica), compiled around 200-300 CE, where it was classified as a superior herb, indicating its high value and relative safety. In TCM, Astragalus root was traditionally used to strengthen the ‘Qi’ (vital energy), particularly of the spleen and lungs, to strengthen resistance to disease, promote tissue regeneration, and increase vitality. It was commonly used for fatigue, weakness, frequent colds, shortness of breath, and poor appetite. The modern scientific study of ononin began in the mid-20th century, with its isolation and characterization from various plant sources.

The structure of ononin was elucidated as formononetin-7-O-glucoside, distinguishing it from other isoflavone glycosides by the presence of a methoxy group at the C-4′ position of the aglycone (formononetin) and a glucose moiety attached to the C-7 position via an O-glycosidic bond. Research on ononin’s biological activities expanded significantly in the early 2000s, with studies investigating its antioxidant, anti-inflammatory, neuroprotective, and anticancer properties. The interest in plants containing ononin, particularly Ononis spinosa, red clover, and Astragalus, for various health applications grew during this period, leading to the development of standardized extracts for modern use. In recent decades, research on ononin has expanded to include its potential applications in neurodegenerative diseases, cardiovascular health, bone health, and cancer prevention and treatment.

The unique structure of ononin, with its O-glycosidic bond and methoxylated aglycone, continues to be investigated for its distinct biological activities and potential therapeutic applications. Today, ononin is recognized as one of the bioactive compounds in Ononis spinosa, red clover, and Astragalus, providing a scientific basis for many of their traditional uses while also revealing new potential therapeutic applications based on its unique pharmacological properties.

Preclinical investigations into ononin’s anti-inflammatory effects in various inflammatory conditions, including rheumatoid arthritis, inflammatory bowel disease, and neuroinflammation, Studies on ononin’s neuroprotective effects in models of neurodegenerative diseases, including Alzheimer’s disease and Parkinson’s disease, Investigations into ononin’s renoprotective effects in various kidney injury models, including drug-induced nephrotoxicity and diabetic nephropathy, Research on ononin’s bone-protective effects, especially for postmenopausal osteoporosis, Studies on ononin’s anticancer effects, particularly for lung cancer, breast cancer, and colorectal cancer, Investigations into ononin’s cardiovascular benefits, particularly its effects on oxidative stress, inflammation, and endothelial function, Limited clinical trials evaluating plant extracts containing ononin (such as Ononis spinosa and red clover) for various health conditions, including menopausal symptoms, osteoporosis, and urinary tract disorders