Agrimonia pilosa

Agrimonia pilosa Ledeb. (Hairy Agrimony): A Comprehensive Reference

1. Identity: Botanical Classification, Names, and Forms

Agrimonia pilosa Ledeb., also known as agrimony, Agrimoniae herba, or hairyvein agrimony, belongs to the family Rosaceae. The plant is commonly called hairy agrimony and is a traditional medicinal plant widely used in Eastern Europe and Eastern Asia. In Chinese tradition and in some scientific contexts, it is also referred to as dragon's tooth grass, desiccated grass, and melon herb, and represents the dried above-ground part of the plant from the Rosaceae family, which contains approximately 124 genera and 3,300 species distributed worldwide, with more in the northern temperate zone.

It is a perennial herb that grows in east Asia, central Europe, and the former Soviet Union. The plant is widely distributed in China, Korea, and Japan. There are ten other reported species of Agrimonia plants, including Agrimonia coreana Nakai — a naturally growing species in South Korea — and Agrimonia eupatoria Linn.

Common preparations and forms: The plant is harvested as it comes into flower and can be dried for later usage. The aerial parts of A. pilosa are used as an astringent hemostatic in the Chinese Pharmacopoeia to treat many types of bleeding, including bloody diarrhea, as well as to prevent toxins and reduce swelling in boils and ulcers. Extracts used in research include aqueous decoctions, ethanolic extracts, methanol extracts, and petroleum ether extracts. The Compound Agrimonia Enteritis Capsule, which is based on A. pilosa, is used for treating acute diarrhea caused by spleen deficiency and damp heat, loose stools, malnutrition and tiredness, abdominal distension and abdominal pain, and acute and chronic enteritis.

2. Traditional and Historical Use

Traditional Chinese Medicine (TCM)

In traditional Chinese medicine (TCM), A. pilosa is considered to have hemostasis-regulating, antimalarial, antidiarrheal, detoxification, and complement deficiency-restoring effects. It is astringent and hemostatic, and it is used for treating malaria, preventing dysentery, detoxification, and as a tonic for deficiency.

The classical text ZhongHuaBenCao (Chinese Materia Medica) recorded that the compatibility of A. pilosa with arborvitae leaves and lotus root can treat hemoptysis and hematemesis. Rhizoma Imperatae and Jiaoshanzhi are used for gingival bleeding, and Daji, Mutong, and Rhizoma Rhizoma are administered for hematuria; the National Compendium of Chinese Herbal Medicine documents that a double concentrate of the whole plant is externally used to treat Trichomonas vaginalis infection.

Additional historical information has been gathered from classical books including Ben Cao Tu Jing, A Textual Research on the Name and Reality of Plants, Modern Practical Chinese Medicine, and Zhen Nan Ben Cao. Agrimonia pilosa Ledeb. has traditionally been known for its effective antitumor activity and is currently used in China for cancer therapy.

East Asian and Korean Traditional Use

A. pilosa has been commonly used in Korea and other Asian countries as a natural flavoring agent and to prevent or treat various diseases, such as hemorrhage, chronic fatigue syndromes, and liver disorders. The aerial parts have been used as an antiviral and for treating hematochezia, traumatic injury, diarrhea, and leukorrhea in Oriental medicine. The aerial parts of A. pilosa have also been used as a wild vegetable mainly in the mountain villages of Korea.

Eastern European and Central Asian Traditional Use

Hairy agrimony is a traditional medicinal plant widely used in Eastern Europe and Eastern Asia regions. Hairy agrimony has been traditionally used to treat sore throat, abdominal pain, headache, mucoid dysentery, bloody and white discharge, parasites, and eczema.

Documented Traditional Indications (Summary)

• Agrimonia pilosa Ledeb. is primarily used in traditional East Asian medicine to treat abdominal pain, dysentery, and hemostasis.
• It has been used to treat various diseases, such as tumors, trichomoniasis, vaginitis, diarrhea, and dysentery.
• It is astringent and hemostatic, and its traditional uses include treating malaria, preventing dysentery, detoxification, and serving as a tonic for deficiency.
• Agrimonia pilosa Ledeb. has shown remarkable effectiveness in the treatment of various diseases, especially enteritis, gastric ulcers, and gastrointestinal bleeding.

3. Key Constituents and Active Compounds

Over the last few decades, approximately 252 compounds have been separated from A. pilosa (APL), including flavonoids, volatile oils, tannins, phenols, phloroglucinol, pentacyclic triterpenoids, isocoumarins, lignans, organic acids, and others. Flavonoids, phenols, and tannins have many pharmacological activities and are considered to be the main active constituents of APL.

Flavonoids

Total A. pilosa flavonoids inhibit α-glucosidase activity; the active components are luteolin, quercetin, vitexin, and isovitexin, among which quercetin has the highest activity and noncompetitively inhibits α-glucosidase. Catechin, luteolin, quercetin, quercitrin, hyperoside, rutin, and luteolin 7-O-β-glucoside exhibit significant DPPH free radical-scavenging activity, with IC₅₀ values of 5.06, 7.29, 4.36, 7.12, 6.34, 6.36, and 8.12 μM, respectively. The pharmacological activity of A. pilosa may be attributed primarily to its phenolic compounds agrimoniin, catechin, quercetin, and rutin. Additional isolated flavonoids include quercetin-7-O-β-d-rhamnoside, apigenin-7-O-β-d-glucopyranoside, kaempferol-7-O-β-d-glucopyranoside, kaempferol, and apigenin.

Phloroglucinol Derivatives

Agrimol A, B, C, D, and E were isolated from A. pilosa petroleum ether extract by the Shanghai Institute of Materia Medica and the Shanghai Fourteenth Pharmaceutical Factory in 1975. The phenolic compounds agrimol F and G were isolated from A. pilosa ethyl ether extract by Yamaki et al. Agrimophol and pseudoaspidin were isolated from the petroleum ether extract of A. pilosa rhizomes by Pei et al. Agrimophol (agrimol A, B, and C), the main constituent of A. pilosa Ledeb., has been widely investigated for the treatment of osteosarcoma, pancreatic carcinoma, prostatic carcinoma, and lung carcinoma.

Tannins and Organic Acids

Tannins and organic acids in A. pilosa mainly include potentillin, pedunculagin, casuarinin, isovanillic acid, and protocatechuic acid, which exhibit antitumor, anti-inflammatory, and free radical-scavenging activities. Agrimoniin (C₈₂H₅₄O₅₂), with a molecular weight of 1,871.282, is a dimeric ellagitannin responsible for the antitumor activity of A. pilosa Ledeb. Agrimoniin is a polyphenol from the group of tannins with antioxidant and anticancer activities.

Triterpenes

Triterpenes are the main chemical constituents of A. pilosa. Several bioactive monomers, such as corosolic acid, euscaphic acid, ursolic acid, and pomolic acid, are also found in A. pilosa. Most of A. pilosa triterpenes promote insulin sensitivity, improve glucose metabolism, and reduce oxidative stress in vitro, indicating that they have potential for the development of antidiabetic drugs.

Isocoumarins

The phytochemical study of A. pilosa based on α-glucosidase inhibitory (AGI) activity led to the isolation of four isocoumarins: agrimonolide (1), agrimonolide-6-O-β-d-glucopyranoside (2), desmethylagrimonolide (3), and desmethylagrimonolide-6-O-β-d-glucopyranoside (4), and four flavonoids: luteolin (5), quercetin (6), vitexin (7), and isovitexin (8). The four isocoumarins were isolated as α-glucosidase inhibitors for the first time.

4. Mechanisms of Action

Anti-inflammatory Mechanisms

Ethanolic extracts of Agrimonia pilosa Ledeb. have been shown to have anti-inflammatory and anti-allergic activities inhibiting inflammatory cytokines (IL-1β, IL-4, IL-6) and interferon-β (INF-β) production in induced RAW 246.7 cells. An ethanolic extract of Agrimonia pilosa Ledeb. was found to inhibit NO and PGE₂ production by downregulating iNOS and COX-2 expression. The ethanol extract also presented immunomodulatory activity, decreasing the upregulation of gene expression of COX-2, iNOS, IL-6, TNF-α, and NF-κB in lipopolysaccharide (LPS)-induced peripheral blood mononuclear cells.

Antitumor Mechanisms

The anti-cancer effect of A. pilosa can be attributed to the polyphenol Agrimol B; the effect on cancer cells may originate from its effect on c-MYC, SKP2, and p27, as observed by immunoblotting and immunofluorescence. Agrimol B enriches prostate and lung cancer cells in the G0 phase (a reversible quiescent state outside the cell cycle) and affects key regulators that control the G0 state.

Agrimol B (Agr) was found to increase ROS generation, block Bcl-2 expression, and increase Caspase-3 and Bax expression to promote cancer cell apoptosis. Agrimol B inhibited colon carcinoma progression by promoting oxidative stress and cell apoptosis by blocking the PGC-1α/NRF1/TFAM signaling pathway.

The anticancer action of agrimoniin is associated with the activation of mitochondria-dependent apoptosis; it exerts a direct influence on different mitochondrial functions, including the induction of the mitochondrial permeability transition pore (MPTP) as the primary mechanism of mitochondria-dependent apoptosis. The effects of agrimoniin on mitochondria are possibly based on the interaction of its numerous hydroxyl groups with adenylate translocase, a key regulator of both MPTP and oxidative phosphorylation.

Antidiabetic Mechanisms

Four isocoumarins — agrimonolide, agrimonolide-6-O-β-D-glucopyranoside, desmethylagrimonolide, and desmethylagrimonolide-6-O-β-D-glucopyranoside — were found to be α-glucosidase inhibitors. The endogenous glucose-inhibitory activity of agrimonolide is related to the inhibition of phosphoenolpyruvate carboxykinase, which is the rate-limiting enzyme in the gluconeogenesis pathway (IC₅₀, 8.3 μmol/L). In insulin-resistant cells, agrimonolide improves insulin sensitivity and promotes insulin-mediated glycogen synthesis.

Anthelmintic Mechanisms

Agrimophol inhibits glycogen decomposition in tapeworm by directly coming into contact with the tapeworm's body, thereby inhibiting tapeworm aerobic and anaerobic metabolism. Agrimol G destroys the parasite cuticle when incubated with adult Haemonchus parasites for 3 hours, and elicits a killing effect on Haemonchus parasites by inhibiting microtubule aggregation.

Coagulation Modulation

APL aqueous extracts at 4 g/L significantly prolonged whole blood clotting time and activated partial thromboplastin time, shortened prothrombin time, decreased activities of coagulation factor VIII, IX and XI, and levels of platelet aggregation and fibrinogen receptor expression. Coagulation factor VII activity and blood viscosity were increased after extract treatment, and the effects of APL extracts were in a concentration-dependent manner (0–80 g/L). The results suggest that APL aqueous extracts have a total anticoagulant activity, while exhibiting opposite effects with greater anticoagulant activity than procoagulant activity.

5. Scientific Evidence by Area of Use

5.1 Oncology / Antitumor Activity

Evidence base: Predominantly in vitro (cell-line) and some in vivo (animal xenograft) studies. No clinical trials in humans have been reported to date.

Extracts of A. pilosa exhibit antitumor effects in a concentration-dependent manner against several tumor cell lines, including MKN-45 human gastric cancer cells, HepG2 human hepatoma cells, U266 human multiple myeloma cells, MCF-7 human breast cancer cells, A549 human non-small cell lung cancer cells, and HeLa cells, with IC₅₀ values of 127.50, 53.31, 202.10, 206.80, 54.17, and 170.40 μg/mL, respectively.

The pure compounds Agrimol B and Agrimoniin of APL have in vitro anticancer and antitumor activity that inhibits the growth of cancer and tumor cells. The oral bioavailability of Agrimol B is 3.01, and the oral bioavailability of Agrimoniin is 19.38.

A study reported that A. pilosa methanol extract inhibited the invasion of cancer cells through inactivation of ERK and JNK in HT1080 cells. Oral administration of APL or Agrimol B has also been shown to reduce the growth of xenografts in animal prostate cancer cells, thereby demonstrating anti-cancer effects.

Although several studies have examined and isolated chemical constituents from this plant, little evidence exists supporting the antitumor activity of individual molecules identified in these studies. Detailed mechanisms of antitumor activity of A. pilosa Ledeb. have not been fully elucidated. Overall, evidence at this stage is preliminary and confined to preclinical models.

5.2 Metabolic / Antidiabetic Activity

Evidence base: In vitro enzyme inhibition studies and in vivo mouse models; no robust human clinical trials for A. pilosa specifically.

In Chinese traditional medicine, A. pilosa Ledeb. exhibits a great effect on treatment of type 2 diabetes mellitus (T2DM); considering that T2DM is correlated with postprandial hyperglycemia and oxidative stress, researchers investigated the α-glucosidase inhibitory activity and the antioxidant activity of flavonoid compound (FC) and triterpenoid compound (TC) from APL. The maximum inhibition was found to be 94.5% and 87.5% at 41.67 μg/mL of FC and TC, respectively. Compared with FC (IC₅₀ = 8.72 μg/mL), TC exhibited stronger efficiency with an IC₅₀ of 3.67 μg/mL.

Isocoumarins — compound 1 (agrimonolide) and 3 (desmethylagrimonolide) — showed strong α-glucosidase inhibitory activities with IC₅₀ values of 24.2 and 37.4 µM, respectively. In kinetic analysis, isocoumarins (compounds 1 and 3) showed non-competitive inhibition, whereas flavonoid compound 6 (quercetin) showed competitive inhibition.

In the mouse, a water Agrimonia pilosa Ledeb. extract alleviated metabolic syndrome symptoms while increasing the levels of adiponectin, an anti-inflammatory protein produced in adipose tissue. Evidence remains preliminary; no human clinical trials have been published for A. pilosa specifically in diabetes management.

5.3 Blood Coagulation and Hemostasis

Evidence base: In vitro hematological experiments; no controlled human clinical trials identified.

APL has showed anticoagulant and antithrombotic activities in some studies, whereas its actual effects on blood coagulation were initially unclear. A study was designed to observe the in vitro effects of APL aqueous extracts on blood coagulation and to investigate the underlying mechanisms. Studies were divided into four groups: 0, 4, 20, and 80 g/L of APL aqueous extracts mixed with plasma or whole blood samples. Clotting time of whole blood, plasma coagulation tests, activities of plasma coagulation factors, plasma calcium ion, platelet aggregation test, platelet fibrinogen receptor, and blood viscosity were measured. The in vitro results demonstrated a concentration-dependent, predominantly anticoagulant profile, which contrasts with the plant's traditional use as a hemostatic agent — a discrepancy that remains scientifically unresolved. These are in vitro data only and cannot be directly extrapolated to clinical use.

5.4 Antimicrobial and Anthelmintic Activity

Evidence base: In vitro and limited in vivo preclinical data; limited clinical data exist specifically for agrimophol as a tapeworm expellant.

Several phloroglucinol derivatives present in A. pilosa have antibacterial activities; for example, agrimol C, agrimol F, agrimol G, and agrimophol completely inhibit the growth of methicillin-resistant Staphylococcus aureus, Bacillus cereus, and Gardnerella species. However, experiments on the antibacterial activity of phenolic components of A. pilosa were performed in 1988, and the specific mechanism has not been clarified. Given the possibility of bacterial variation and drug resistance, the antibacterial activity of phenolic compounds in A. pilosa should be further investigated.

Agrimophol is the active ingredient of agrimony from the dry part above the ground of Agrimonia pilosa Ledeb., which has excellent anthelmintic effect, and can also be used for treating trichomonas enteritis, vaginitis, and other diseases. Clinical application and pharmacological studies have shown that agrimony root has a broad spectrum of anthelmintic effect with high activity and low toxicity. Agrimonia essential oil (at concentrations of 10, 50, and 100 μg/mL) shows a dose-dependent inhibitory activity on Leishmania promastigote and intracellular amastigote forms in vitro, with no toxic effects on host cells at these concentrations.

5.5 Antiviral Activity

Evidence base: In vitro cell-culture studies and in vivo mouse studies; no human clinical trials published.

Agrimonia pilosa ethanol extract (APEE) exhibited potent antiviral activity against SARS-CoV-2 with an IC₅₀ of 1.1 ± 0.03 µg/mL. Its mechanism of action included virucidal activity and inhibition of viral adsorption.

Agrimonia pilosa (AP), Galla rhois (RG), and their mixture (APRG64) strongly inhibited SARS-CoV-2 by interfering with multiple steps of the viral life cycle including viral entry and replication. Among 12 components identified in APRG64, three displayed strong antiviral activity: ursolic acid (1), quercetin (7), and 1,2,3,4,6-penta-O-galloyl-β-d-glucose (12). Molecular docking analysis showed these components to bind potently to the spike receptor-binding-domain (RBD) of SARS-CoV-2 and its variant B.1.1.7.

The mixture of A. pilosa and gallnut extract (APRG64) at a 6:4 ratio significantly inhibits the expression of HCV core 1b and NS5A proteins at a concentration of 5 μg/mL in vitro, with no obvious cytotoxicity; the inhibitory activity of luteolin was the most significant (P < 0.01). However, only in vitro experiments have been conducted, and antiviral activity of a single A. pilosa extract is significantly weaker than that of the mixture. All antiviral findings are from in vitro and animal studies; clinical evidence in humans is absent.

5.6 Anti-inflammatory and Analgesic Activity

Evidence base: In vitro and in vivo (animal) studies only; no human clinical trials reported.

Crude extracts and their isolates show a broad panel of pharmacological activities including anti-cancer, anti-microbial, antivirus, anti-oxidant, anti-inflammation, anti-diabetes, anti-osteosarcoma, anti-aging, anti-nociception, anti-adipogenesis, anti-leishmaniasis, estrogenic-like activity, neuroprotective and hepatoprotective activities, and vascular relaxation. The anti-inflammatory and analgesic properties are mechanistically linked to suppression of iNOS/COX-2 expression and inhibition of NF-κB pathway signaling, though these findings derive from cell and animal models.

5.7 Antioxidant Activity

Evidence base: In vitro free radical scavenging assays; no clinical studies.

Aqueous extracts of the aerial parts of A. pilosa have shown antiviral activity against hepatitis B viruses as well as anticoagulant activity. Water-alcoholic extracts of A. pilosa leaves demonstrated antioxidant and anti-inflammatory activities. The antioxidant effect is attributed primarily to flavonoids, with DPPH radical-scavenging IC₅₀ values in the low micromolar range as noted above. All findings are from in vitro assays.

5.8 Neuroprotective and Acetylcholinesterase Inhibitory Activity

Evidence base: In vitro enzyme inhibition; preliminary only.

Pharmacological studies on extracts prepared from the aerial parts of A. pilosa demonstrated broad biological properties, including antihemorrhagic, antiplatelet, antioxidant, nitric oxide scavenging, acetylcholinesterase inhibitory, and α-glucosidase inhibitory activities. Ellagitannin and flavonoid constituents have been studied for protein tyrosine phosphatase and acetylcholinesterase inhibitory activities. Evidence is entirely preclinical.

6. Dosage Forms and Reported Study Dosages

The following dosages and forms are those specifically reported in published scientific studies. They are not therapeutic recommendations.

• Aqueous extracts (in vitro hematological study): Studies used groups of 0, 4, 20, and 80 g/L of APL aqueous extracts mixed with plasma or whole blood samples.
• Agrimonia pilosa ethanol extract (APEE) against SARS-CoV-2: The ethanol extract exhibited potent antiviral activity with an IC₅₀ of 1.1 ± 0.03 µg/mL.
• APRG64 mixture against SARS-CoV-2 (in vitro): Cells were treated with AP, RG, or APRG64 at 0.1 or 0.5 µg/mL for 2 hours and then infected with SARS-CoV-2 at 0.01 multiplicity of infection.
• Agrimol B (in vitro cancer, Agr concentrations): Cells were treated with different concentrations of Agr (0, 144, 288, and 576 nM), and cell viability, migration rate, proliferation rate, and apoptosis rate of human colon cancer HCT116 cells were determined.
• Alpha-glucosidase inhibitory study (flavonoid and triterpenoid fractions): The maximum inhibition was found to be 94.5% and 87.5% at 41.67 μg/mL of FC and TC respectively; compared with FC (IC₅₀ = 8.72 μg/mL), TC exhibited stronger efficiency with an IC₅₀ of 3.67 μg/mL.
• Agrimonolide (isocoumarin) α-glucosidase inhibition: Agrimonolide and desmethylagrimonolide showed strong α-glucosidase inhibitory activities with IC₅₀ values of 24.2 and 37.4 µM, respectively.
• Essential oil (anti-Leishmania, in vitro): Agrimonia essential oil at concentrations of 10, 50, and 100 μg/mL showed dose-dependent inhibitory activity on Leishmania promastigote and intracellular amastigote forms in vitro.
• Agrimoniin (LD₅₀ in animal toxicity study): The substance was low in toxicity, with LD₅₀ equal to 100 mg/kg and over 1,000 mg/kg when administered to mice peritoneally and orally, respectively.

7. Body Systems and Health Areas of Association

• Gastrointestinal system: Traditional use and some clinical formulations target enteritis, gastric ulcers, gastrointestinal bleeding, dysentery, and diarrhea. The Compound Agrimonia Enteritis Capsule is used for treating acute and chronic enteritis, loose stools, abdominal distension and pain.
• Hematological / cardiovascular system: The plant is used as a hemostatic agent in TCM and has demonstrated in vitro dual coagulation-modulating effects.
• Oncology: In vitro and animal evidence for antitumor activity against multiple cancer lines; currently used in China as a component of cancer-related formulas.
• Metabolic / endocrine system: α-glucosidase inhibitory and insulin-sensitizing activities demonstrated in vitro and in vivo.
• Immune system / infectious disease: Antiviral, antibacterial, and antiparasitic activities documented in preclinical studies.
• Nervous system: Acetylcholinesterase inhibitory activity of ellagitannins and flavonoids demonstrated in vitro.
• Reproductive / genitourinary system: Traditional use for trichomoniasis and vaginitis; some estrogenic-like activity reported in preclinical studies.

8. Safety Considerations

A very limited range of A. pilosa toxicity tests have been documented regarding their genetic toxicity, acute, and sub-chronic oral toxicity, and systemic safety including the respiratory system, central nervous system, and cardiovascular system. As of at least 2018, no comprehensive studies on the toxicity of AP as a whole plant had been reported in the scientific literature.

For the isolated compound agrimoniin, animal data showed low toxicity, with LD₅₀ equal to 100 mg/kg (peritoneal) and over 1,000 mg/kg (oral) in mice.

Agrimophol-specific adverse effects: Fewer adverse effects were observed with agrimophol; occasional nausea, vomiting, dizziness, cold sweat, or diarrhea have been reported, and a collapse reaction has been described after a half month of use.

Dual coagulation effects — a notable safety concern: APL has shown both anticoagulant and antithrombotic activities in some studies, while its actual effects on blood coagulation are concentration-dependent and complex. The opposite procoagulant and anticoagulant effects observed at different concentrations in vitro suggest that concurrent use with anticoagulant or antiplatelet drugs could produce unpredictable interactions; however, this has not been formally studied in humans.

Insufficient clinical data: Most prescriptions for Agrimonia pilosa Ledeb. are empirical and lack rigorous clinical observation. The toxicology, standardized clinical studies, nature of active ingredients, pharmacokinetics, mechanism, and metabolism of Agrimonia pilosa Ledeb. should be deepened, especially through clinical trials, to ensure clinical safety of its use for further research.

Ca²⁺ channel interaction: Through patch-clamp recording, previous studies reported that Agrimonia plant extracts inhibit the function of Ca²⁺ release-activated Ca²⁺ channels (CRACs). The clinical implications of this finding for users of calcium channel-modulating drugs have not been formally evaluated.

Pharmacopoeia status: A. pilosa is still not listed in many countries and other official pharmacopoeias. The plant is, however, listed in the Pharmacopoeia of the People's Republic of China.

9. Overall Assessment of Evidence Quality

In vitro and in vivo results have successfully explained the pharmacological mechanisms of A. pilosa constituents. More bioassay-guided phytochemical and clinical studies are necessary. Although recent studies have isolated numerous active constituents and investigated their effects, the medicinal utility of this herb is not yet fully explored.

The totality of current evidence for Agrimonia pilosa is almost entirely preclinical (in vitro cell-line and in vivo animal studies). In vitro and in vivo results have successfully explained pharmacological mechanisms of A. pilosa constituents; however, more bioassay-guided phytochemical and clinical studies are necessary. No adequately powered, randomized human clinical trials have been published for any of the major claimed therapeutic indications. The traditional use in TCM and folk medicine across East Asia and Eastern Europe provides a long historical record, but most prescriptions for Agrimonia pilosa Ledeb. are empirical and lack rigorous clinical observation.

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