Synonym name: Apigenin-7-O-neohesperidoside; Rhoifoloside
Catalogue No.: BP1215
Cas No.: 17306-46-6
Formula: C₂₇H₃₀O₁₄
Mol Weight: 578.523
Botanical Source: Chorisia spp., Citrus aurantium (Seville orange), Rhus succedanea and many other plant spp.

Purity: 95%~99%
Analysis Method: HPLC-DAD or/and HPLC-ELSD
Identification Method: Mass, NMR
Packing: Brown vial or HDPE plastic bottle
Can be supplied from milligrams to grams.

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

Description:

Rhoifolin possesses a variety of significant biological activities including antioxidant, anti-inflammatory, antimicrobial, hepatoprotective and anticancer effects, it may be beneficial for diabetic complications through its enhanced adiponectin secretion, tyrosine phosphorylation of insulin receptor-β and GLUT4 translocation. Rhoifolin has a radioprotective effect against radiation-induced decrease of blood platelets and cardiac biochemical lesions in whole body irradiated mice.

References:

Evid Based Complement Alternat Med. 2011;2011:624375.    

Insulin-Mimetic Action of Rhoifolin and Cosmosiin Isolated from Citrus grandis (L.) Osbeck Leaves: Enhanced Adiponectin Secretion and Insulin Receptor Phosphorylation in 3T3-L1 Cells. 

Citrus grandis (L.) Osbeck (red wendun) leaves have been used in traditional Chinese medicine to treat several illnesses including diabetes. However, there is no scientific evidence supporting these actions and its active compounds. 

METHODS AND RESULTS:

Two flavone glycosides, Rhoifolin and cosmosiin were isolated for the first time from red wendun leaves and, identified these leaves are rich source for Rhoifolin (1.1%, w/w). In differentiated 3T3-L1 adipocytes, Rhoifolin and cosmosiin showed dose-dependent response in concentration range of o.oo1-5 μM and 1-20 μM, respectively, in biological studies beneficial to diabetes. Particularly, Rhoifolin and cosmosiin at 0.5 and 20 μM, respectively showed nearly similar response to that 10 nM of insulin, on adiponectin secretion level. Furthermore, 5 μM of Rhoifolin and 20 μM of cosmosiin showed equal potential with 10 nM of insulin to increase the phosphorylation of insulin receptor-β, in addition to their positive effect on GLUT4 translocation. 

CONCLUSIONS:

These findings indicate that Rhoifolin and cosmosiin from red wendun leaves may be beneficial for diabetic complications through their enhanced adiponectin secretion, tyrosine phosphorylation of insulin receptor-β and GLUT4 translocation.    

Zhongguo Zhong Yao Za Zhi. 2014 Aug;39(16):3094-101.    

[Chemical constituents from Callicarpa nudiflora and their cytotoxic activities].[Pubmed: 25509294]    

The chemical consitituents from cytotoxic fraction of the Callicarpa nudiflora extract were isolated and purified by a combination of HP-20 macroporous resin, silica gel and Sephadex LH-20 column chromatographies. 

METHODS AND RESULTS:

The structures were elucidated on the basis of the spectroscopic data and comparison of their spectroscopic data with reported data. The cytotoxicity was evaluated by the MTT assay. The 50% and 70% EtOH elutions of EtOH-extract showed significant cytotoxic activities, leading to the isolation of twelve compounds, which were identified as luteoloside(1), lutedin-4'-O-β-D-glucoside(2), 6-hydroxyluteolin-7-O-β-glucoside(3), lutedin-7-O-neohesperidoside(4), Rhoifolin (5), luteolin-7, 4'-di-O-glucoside (6), forsythoside B (7), acteoside (8), alyssonoside (9), catalpol(10), nudifloside(11), and leonuride(12). Compounds 3-6, 10 and 12 were isolated from this genus for the first time, and compound 9 was isolated from this plant for the first time. The cytotoxicity assay demonstrated that flavonoids 1-6, in various concentrations, showed monolithic proliferation inhibitory activities against Hela, A549 and MCF-7 cell lines. Compounds 3, 5 and iridoid glycoside 11 possessed higher cytotoxicacivities. 

CONCLUSIONS:

In short, flavonoids are the main components of cytotoxic extract from C. nudiflora, while phenylethanoid glycosides are the predominant ingredient but inactive to cancer cell lines. In addition, the minor iridoid glycoside expressed weak cytotoxic activity.    

Pharmacology & Clinics of Chinese Materia Medica, 2012(6):18-20.    

Study on the protective effect of rhoifolin against hepatic injury induced by triptolide   

Study on the protective effect of Rhoifolin against hepatic injury induced by triptolide.

METHODS AND RESULTS:

The protective effects of Rhoifolin at the concentration of 75,150,300μg/ml,on the in vitro damage of human L02 liver cells injured by triptolide at the concentration of 40μg/ml were evaluated with cell survival rates,which were determined by MTT assay.Sixty mice were randomly divided into six groups as follows: The drug groups mice were treated with Rhoifolin for six day at the dose of 140,70,35mg/kg,respectively.The normal control group and the model control group took equivalent amount of distilled water by the same route.And mice in the positive control group were treated with 70mg/kg diammonium glycyrrhizinate.After the last administration,all groups except the normal control were administered with triptolide at dose of 0.625mg/kg by gavage to induce acute hepatic injury.Thereafter,the contents of ALT,AST in serum,the activities of SOD,MDA,GSH-Px and GST in the liver were measured and the hepatic histological changes were observed by optical microscope. The cell survival rates of human L02 liver cells injured by triptolide and then treated with Rhoifolin at the concentration of 75,150,300μg/ml were significantly increased.The contents of ALT,AST in serum were significantly decreased,the content of MDA in the liver was also significantly lowered,and the activities of SOD,MDA,GSH-Px and GST in liver were significantly improved after preventive treatment with Rhoifolin at the dose of 140,70,35mg/kg.Rhoifolin can ameliorated the hepatic pathological changes.

CONCLUSIONS:

The protective effect of Rhoifolin against hepatic injury induced by triptolide was significant in vitro and in vivo.  

J. Appl. Pharmaceut. Sci., 2012, 2(8):74-9.    

Anti-inflammatory effect of Apigenin-7-neohesperidoside (Rhoifolin) in carrageenin-induced rat edema model   

Flavonoids are normal constituents of the human diet and are known for a variety of biological activities. They have been reported to bring benefits in lowering inflammation and oxidative stress.

METHODS AND RESULTS:

The present investigation was performed first, to evaluate the anti-inflammatory activity of Rhoifolin and second, to search for the possible contributing mechanisms for this hypothesized effect. Rhoifolin caused a time and reverse dose dependent reduction of carrageenin-induced rat paw oedema. Following 4 hr of treatment, Rhoifolin at doses 2.5, 25 & 250 mg/kg caused a significant inhibition of rat paw edema volume by 14, 25 & 45 % respectively in comparison to the control group (74%). In addition to significantly abrogating prostaglandin E2 level, increasing doses of Rhoifolin significantly diminished the TNF-a release in the inflammatory exudates. In the same animal model, Rhoifolin increased the total antioxidant capacity in a reverse dose order, with the highest capacity obtained with the lowest dose tested. 

CONCLUSIONS:

This study demonstrates for the first time the effectiveness.    

 HNMR of Rhoifoloside

HPLC of Rhoifolin

​

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

Introduction/Overview

Diabetes, inflammatory diseases, osteolysis, malignant tumors and other major diseases seriously threaten human health, and the demand for new therapeutic drugs is increasingly urgent. Natural products have always been an important source of innovative drug discovery due to their structural diversity and rich biological activity. Flavonoids, as one of the major classes, are widely distributed in the plant kingdom and exhibit various pharmacological activities including antioxidant, anti-inflammatory, and anti-tumor effects. Rhoifolin, a flavonoid carbon glycoside with a unique pharmacological spectrum, has attracted much attention in recent years because of its remarkable activities in anti diabetes, anti-inflammatory, anti osteolysis and anti-tumor. It is mainly derived from plants in the Rutaceae family, such as wild lacquer（Rhus succedanea L. Separated from others. This article aims to systematically review the chemical structure, plant origin, pharmacological activity, molecular mechanism of action, pharmacological characteristics, and clinical application potential of wild lacquer glycoside, in order to provide comprehensive scientific references for the in-depth research and development of this compound.

Chemical structure and physicochemical properties

Wild lacquer tree glycoside (CAS number: 17306-46-6), also known as apigenin-7-O-neohesperidoside, is a flavonol carbon glycoside. Its molecular formula is C27H30O14 and its molecular weight is 578.5230. Its basic skeleton is apigenin, which is a 5,7,4 '- trihydroxyflavone. A disaccharide, neohesperidose (also known as rhamnose - (α 1 → 2) - glucose), is connected to its 7th hydroxyl group through a glycosidic bond. This unique glycosylation structure has a decisive impact on its water solubility and biological activity.

From the analysis of physical and chemical properties, the theoretical lipid water partition coefficient (LogP) of wild lacquer glycoside is about -0.1675, indicating its good hydrophilicity. Its topological polar surface area (TPSA) is as high as 228.9700 Å ², mainly attributed to the numerous oxygen atoms and sugar moieties in the molecule. The predicted value of its water solubility is about 2.2997 mg/mL, which belongs to the soluble range. These parameters collectively determine its poor membrane permeability, especially the predicted "low" ability to penetrate the blood-brain barrier, which limits its direct potential for central nervous system related diseases. In the preliminary safety screening, the hERG inhibition risk is "no", indicating a low risk of cardiac toxicity; The Ames test value is 0.6, indicating a low risk of mutagenicity, but further in vitro and in vivo experiments are needed to confirm.

Plant sources and extraction methods

Wild lacquer glycosides are mainly found in plants of the Rutaceae and Anacardiaceae families. Its name comes from the plant wild lacquer（Rhus succedanea L. It is one of its main sources. In addition, the peel and leaves of citrus fruits such as grapefruit and lime, as well as various medicinal plants such as Citrus aurantium(Lime)Agrimonia pilosa It has also been found in plants such as Longya Grass.

The extraction of wild lacquer glycoside from plant materials is usually carried out using organic solvent extraction method. Common solvents include methanol, ethanol, or their aqueous solutions. In order to improve extraction efficiency, modern extraction techniques such as ultrasound assisted extraction (UAE), microwave-assisted extraction (MAE), and pressurized liquid extraction (PLE) have been widely used. For example, using a 70% -80% ethanol aqueous solution to extract dried plant powder under ultrasound conditions can effectively improve the yield of wild lacquer glycoside. After decompression and concentration, the extract is preliminarily enriched and decolorized by macroporous adsorption resin (such as AB-8, D101) column chromatography, and further separated and purified by silica gel column chromatography, Sephadex gel column chromatography, and high-performance liquid chromatography (HPLC) preparation chromatography, finally obtaining high-purity laccase monomer. The optimization of extraction process needs to comprehensively consider factors such as solvent polarity, temperature, time, solid-liquid ratio, and plant parts (leaves, skin, fruits).

Pharmacological activity research

Numerous in vitro and in vivo pharmacological studies have shown that wild lacquer glycoside has multiple biological activities.

1. Antidiabetic activity: Rhus verniciflorin showed a clear improvement in insulin resistance and type 2 diabetes models. Research has shown that it can promote the secretion of adiponectin (a fat factor with insulin sensitizing effects) by adipocytes, enhance the tyrosine phosphorylation level of insulin receptor - β subunit, and activate downstream insulin signaling pathways. More importantly, it can promote the translocation of glucose transporter 4 (GLUT4) from the cytoplasm to the cell membrane, directly accelerating the uptake and utilization of glucose by peripheral tissues such as fat and muscle, and reducing blood sugar levels.

2. anti-inflammatory activity The anti-inflammatory effect of wild lacquer glycoside is one of its most prominent pharmacological properties. In lipopolysaccharide (LPS) - induced macrophage inflammation models and various animal inflammation models (such as mouse ear swelling and arthritis models), wild lacquer glycoside can significantly inhibit the production and release of inflammatory mediators. It can downregulate the expression of nitric oxide synthase (NOS2/iNOS) and cyclooxygenase-2 (PTGS2/COX-2), reduce the production of nitric oxide (NO) and prostaglandin E2 (PGE2). Meanwhile, it can effectively inhibit the expression of key pro-inflammatory cytokines such as tumor necrosis factor - α (TNF - α) and interleukin-6 (IL-6).

3. Anti osteolysis and inhibition of osteoclastogenesis activity In diseases such as aseptic loosening after artificial joint replacement surgery, worn titanium particles can induce inflammatory bone resorption. Wild lacquer tree glycoside can effectively improve bone destruction under titanium particle stimulation. The mechanism is to inhibit osteoclast differentiation induced by receptor activator of nuclear factor kappa B ligand (RANKL). Osteoclasts are the main cells responsible for bone resorption, and their excessive activation leads to bone resorption.

4. Antitumor activity: Rhus verniciflorin showed dose-dependent cytotoxicity to a variety of human cancer cell lines, including breast cancer, lung cancer, liver cancer, colon cancer, etc. It can induce cancer cell cycle arrest (such as G2/M phase arrest) and promote its apoptosis. In addition, the study suggests that it may exert its anti metastatic potential by inhibiting tumor cell migration and invasion.

Mechanism of action and molecular targets

The multiple pharmacological activities of wild lacquer glycoside stem from its regulation of multiple key signaling pathways, and its target network is complex and precise.

• The core pathway of anti-inflammatory effect The anti-inflammatory effect of wild lacquer tree glycoside is mainly achieved by inhibiting two core inflammatory signaling pathways, nuclear factor kappa B (NF - κ B) and signal transduction and transcription activator 3 (STAT3).

  • NF - κ B pathway In the resting state, NF - κ B (mainly composed of RELA/p65 subunits) binds to the inhibitory protein I κ B and exists in the cytoplasm. Inflammatory stimuli such as LPS and TNF - α activate the I κ B kinase complex (IKK, with IKBKB/IKK β being the key catalytic subunit), leading to phosphorylation and degradation of I κ B, thereby releasing NF - κ B into the nucleus and initiating transcription of inflammatory genes such as IL-6, TNF, NOS2, COX-2, etc. Wild lacquer tree glycoside has been proven to inhibit the activation of IKK β, prevent the degradation of I κ B and the nuclear translocation and DNA binding activity of NF - κ B p65, and block the inflammatory cascade reaction upstream.
  • STAT3 pathway After binding to cytokines such as IL-6 and their receptors, JAK kinase is activated, leading to STAT3 phosphorylation. Phosphorylated STAT3 forms dimers and enters the nucleus, serving as transcription factors to drive inflammation and tumor related gene expression. Wild lacquer glycoside can inhibit the phosphorylation of STAT3, thereby interrupting this pathway.
  • Other targets Research has also shown that wild lacquer glycoside can inhibit the activation of NLRP3 inflammasome, reduce the activity of caspase-1 (CASP1), and thus reduce the release of mature inflammatory factors such as IL-1 β. In addition, it may also have regulatory effects on transient receptor potential vanillic acid subtype 1 (TRPV1) and ANKYRIN 1 (TRPA1) channels, which play important roles in pain and neurogenic inflammation.

• Mechanism of Anti Osteolysis Action During osteoclast differentiation, RANKL binds to its receptor RANK, activating downstream NF - κ B and mitogen activated protein kinase (MAPK, including ERK, JNK, p38) pathways, ultimately inducing osteoclast specific gene expression. Wild lacquer tree glycoside can simultaneously inhibit the activation of NF - κ B and MAPK pathways induced by RANKL, thereby inhibiting the differentiation of osteoclast precursor cells into mature osteoclasts and reducing bone resorption.

• Anti diabetes mechanism: In addition to the aforementioned promotion of adiponectin secretion, enhancement of insulin receptor phosphorylation and GLUT4 translocation, its anti-inflammatory effect itself also helps to improve insulin resistance, because chronic low-grade inflammation is an important pathological basis of type 2 diabetes. By inhibiting inflammatory pathways such as IKK β/NF - κ B and JNK, kaempferol can alleviate the interference of inflammation on the insulin signaling pathway.

• Potential mechanisms of anti-tumor treatment Its anti-tumor activity is closely related to inducing cell apoptosis, which may involve activating the mitochondrial apoptosis pathway (regulating the Bcl-2/Bax ratio and activating Caspase-3/9). Meanwhile, its inhibition of the STAT3 and NF - κ B pathways not only has anti-inflammatory effects, but also blocks the critical roles of these pathways in tumor cell survival, proliferation, invasion, and chemotherapy resistance.

Evaluation of drug properties and pharmacokinetics

Although wild lacquer glycoside exhibits extensive biological activity in vitro, its drug like and pharmacokinetic (PK) properties are key factors determining its successful development as a drug.

• Absorption, distribution, metabolism, excretion (ADME)：

  • absorb As a flavonoid glycoside, wild lacquer glycoside has high polarity (high TPSA, low LogP), which may limit its oral bioavailability. Glycoside forms usually require hydrolytic enzymes (such as β - glucosidase) from the gut microbiota or brush edges of intestinal epithelial cells to hydrolyze them into aglycones (apigenin) and glycosides, before aglycones can be well absorbed through passive diffusion. Therefore, there are individual and species differences in its absorption degree and speed.
  • distribution Predict low blood-brain barrier permeability, mainly distributed in peripheral tissues and organs. The binding rate between it and plasma proteins still needs to be clarified through experimental data.
  • Metabolism Flavonoids are mainly biotransformation in the liver through phase I metabolism (such as hydroxylation and demethylation) and phase II metabolism (glucuronidation and sulfation). Wild lacquer glycoside itself and its aglycone apigenin are substrates for these metabolic enzymes. Metabolites usually have increased polarity, which facilitates excretion.
  • excretion Metabolites are mainly excreted through the kidneys and urine, and some prototype drugs and metabolites may also be excreted through bile and feces.

• Challenges and Strategies in Drug Development：

  1. Balance between water solubility and permeability The current properties tend towards high hydrophilicity and poor membrane permeability. The solubility and transmembrane transport ability can be improved through structural modifications, such as preparing prodrugs (such as esterification, preparation of phospholipid complexes), or developing novel drug delivery systems (such as nanocrystals, liposomes, micelles).
  2. Metabolic stability It is necessary to study its metabolic stability in liver microsomes or liver cells, clarify the main metabolic sites and enzymes, and provide direction for structural optimization.
  3. In vivo efficacy verification Most of the activity data comes from cell models, and there is an urgent need for systematic in vivo pharmacological and dose exploration studies in animal models that are closer to human diseases to confirm their therapeutic window.
  4. safety evaluation Although the initial prediction of hERG and Ames risks is low, a comprehensive preclinical safety evaluation is still needed, including acute toxicity, chronic toxicity, reproductive toxicity, etc.

Clinical application prospects and prospects

The multi-target and multi pathway properties of wild lacquer tree glycosides provide broad prospects for their application in the treatment of complex diseases, but they also face many challenges.

• Potential application directions：

  1. Metabolic diseases As an insulin sensitizer and anti-inflammatory agent, it is expected to be developed as an auxiliary drug or functional food ingredient for the treatment of type 2 diabetes and its complications (such as diabetes nephropathy and neuropathy).
  2. Inflammatory bone disease It has unique potential in the treatment of inflammatory diseases with bone destruction, such as aseptic loosening of artificial joints, rheumatoid arthritis, periodontitis, etc.
  3. Adjuvant anti-tumor therapy Due to its dual anti-inflammatory and apoptosis inducing effects, it may serve as an adjuvant drug for traditional chemotherapy/radiotherapy, enhancing therapeutic efficacy, reducing inflammatory side effects, and overcoming drug resistance.
  4. Local anti-inflammatory and analgesic effects Given its regulatory effect on pain related targets such as TRPV1, it is possible to explore the development of topical formulations for skin inflammation and muscle pain.

• Future research prospects：

  1. In depth mechanism research Using chemical biology methods such as affinity fishing, molecular docking, and kinetic simulations to identify the direct target proteins and create a more accurate target pathway network diagram.
  2. Structural optimization and derivative development Based on its pharmacophore and pharmacological shortcomings, a systematic structural modification was carried out to synthesize a series of derivatives, and candidate compounds with stronger activity, higher bioavailability, and better selectivity were screened.
  3. Research on New Delivery Systems Actively developing biocompatible nanocarrier systems, such as natural product based nanoparticles or exosomes, to enhance their targeting, stability, and in vivo circulation time.
  4. Multi omics integration analysis In animal models or organoid models, transcriptomics, proteomics, metabolomics and other techniques are applied to comprehensively evaluate the systemic biological changes after intervention with wild lacquer glycosides, and discover new biomarkers and potential indications.
  5. Preclinical and clinical translation After completing the pharmacological, pharmacokinetic, and safety evaluations of the system, promote its entry into clinical trials to explore its effectiveness and safety in humans.

Conclusion

As a natural source of flavonoid glycoside, Rhus verniciflorin shows great potential to become a multi target therapeutic drug with its multiple pharmacological activities of anti diabetes, anti-inflammatory, anti osteolysis and anti-tumor. The study of its mechanism of action has delved into key signaling pathways such as NF - κ B, STAT3, MAPK, etc., revealing the scientific connotation of its therapeutic effect through multi-level regulation. However, its inherent pharmaceutical defects, such as low oral bioavailability and fast metabolism, are obstacles that must be overcome in future translational research. Through interdisciplinary strategies such as modern medicinal chemistry, pharmacy, and systems biology, the deep development and utilization of wild lacquer glycoside is expected to transform it from a promising lead compound into a new type of drug for the treatment of metabolic syndrome, chronic inflammation, and bone related diseases, contributing the power of natural products to human health.