Background and overview[1][2]
As a traditional Chinese medicine, Wuyao is one of the important painkillers for warming the stomach, regulating qi and regulating qi. Modern pharmacological research shows that Aegonia has a wide range of pharmacological activities. For example, Aegonia can enhance the secretion of digestive juices, and can also combat abdominal pain caused by clinical use of rhubarb. It has the effects of stimulating and enhancing movement rhythm, can significantly inhibit the formation of ulcers, and can significantly combat abdominal pain. Ethanol-induced cell damage has a cytoprotective effect. Recently, it has been discovered that Wuyao has strong analgesic, anti-inflammatory and significant anti-rheumatic effects. The active ingredient in black medicine is norisoboldine. In the 2005 edition of the “Chinese Pharmacopoeia”, the index components for TLC identification and content determination of Chinese herbal medicine are all lactones. Echinacea lactone is a sesquiterpene component, which is a characteristic component of the herb. It has strong specificity and is suitable as an indicator for identifying the authenticity of the herb. However, agenilide is a fat-soluble component, its decoction rate in water decoction is low, and its content is also low, so it is not the best choice as an indicator for content determination. Therefore, isolating this compound from the original plant is of great significance for quality control indicators, clarifying its pharmacological effects, and finding lead compounds for the development of new drugs.
Structure
Preparation[1]
A method for the separation and purification of norisoboldine monomer, which is characterized in that the norisoboldine monomer is obtained by extracting, extracting and crystallizing from black medicine as raw material, specifically including the following main steps:
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A. Extraction: Crush the black medicine into coarse powder, add ethanol solution with a mass percentage concentration of 95%, the amount added is calculated according to the weight of the medicinal material: the volume of the ethanol solution = 1Kg: (8-12) L, reflux extraction 3 -5 times, 1.5-3 hours each time, filter the extract, combine the filtrate, concentrate to recover ethanol, and obtain a concentrated extract with a relative density of 1.20-1.40 at 50°C for the next step of extraction;
B. Extraction: Dissolve the concentrated extract obtained in step A by adding dilute hydrochloric acid with a mass percentage concentration of 3 pvc stabilizer-4%, adjust the pH to 2-4, filter out the acid aqueous liquid, and extract 3-5 times with chloroform , remove the chloroform liquid, adjust the pH of the acidic aqueous liquid to 8-10 with concentrated ammonia, and then extract 4-6 times with chloroform, combine the chloroform extracts, recover the filtrate to dryness, and obtain yellow norisoboldine crude product;
C. Crystallization: According to the calculation of yellow solid: methanol = 1g: (10-1000) ml, completely dissolve the crude norisoboldine obtained in step B with methanol, ethanol or acetone. After the dissolution is complete, use nanometer Filter through an organic membrane. Add 2-5 times the volume of distilled water to the filtrate for refrigeration crystallization. Repeat crystallization 2 to 3 times according to the specific purity of the crystal until the purity is ≥98%. After drying at 100°C, the norisoboldine monomer can be obtained. product.
Pharmacokinetics[2]
1) After intravenous administration to rats, the concentration of norisoboldine in plasma decreased rapidly, and the plasma concentration was lower than the limit of quantitation (10 ng·mL) after 120 minutes. 5 minutes after simultaneous injection, the metabolite norisoboldine-9- O-α-glucuronide can be detected in the plasma immediately, and norisoboldine-9 The plasma concentration of – O-α-glucuronide is much higher than that of norisoboldine, which shows that the biotransformation of norisoboldine is very rapid in the body. Compared with the prototype drug, the elimination of norisoboldine-9-O-glucuronide in the body is relatively slow, and the plasma concentration does not fall below the limit of quantitation until 12 hours later (25 ng·mL). After intravenous administration in rats, the blood drug concentration data were processed by PKsolution 2TM software according to the non-compartmental model. The pharmacokinetic parameters of norisoboldine and norisoboldine-9-O-α-glucuronide were compared and bentonite was used for statistical testing.
The results show that the pharmacokinetic parameters of norisoboldine and norisoboldine-9-O-α-glucuronide after intravenous injectiont1/2, AUC0t, ke and MRT are statistically significant. According to the metabolic clearance rate of norisoboldine in rat liver microsomes, the clearance rate of norisoboldine in rat liver is 59.4 mL/min/kg, accounting for 10% of the total clearance rate after intravenous administration in rats. rate of 31.9%. Therefore, hepatic metabolism is an important pathway for the elimination of norisoboldine in the body.
2) Pharmacokinetic characteristics after intragastric administration
RatAfter gastric administration, norisoboldine is rapidly absorbed and can be detected after 5 minutes. It reaches the maximum plasma concentration at 23 minutes and is rapidly eliminated thereafter. The plasma concentration is below the limit of quantification after 120 minutes. Just like intravenous administration, 5 minutes after intragastric administration to rats, norepinephrine waves can be detected immediately in the plasma of rats
Erdine-9-O-α-glucuronide, and its plasma concentration is much higher than that of norisoboldine’s prototype drug, so it cannot be ignored. Compared with the prototype drug, the elimination of norisoboldine-9 O-α-glucuronide in the body is relatively slow, and the plasma concentration does not fall below the limit of quantification after 24 hours ( 25 ng·mL). This shows that norisoboldine is absorbed rapidly and has an obvious first-pass effect.
Apply[3]
The total alkaloids of Wuyao can significantly inhibit secondary foot swelling in rats with adjuvant arthritis, and also have a tendency to inhibit primary foot swelling. At the same time, the total alkaloids of Wuyao can also reduce the severity of collagen arthritis in mice, reduce anti-collagen antibodies in serum, and protect bone and joint damage. Its mechanism of action is by preventing lipopolysaccharide-induced NF-κB activation in RAW264.7 cells. and phosphorylation of MAPKs. Norisoboldine, as the main active ingredient in total alkali of aegonia (containing 37.8% in total alkali of aegypti), has pharmacological effects similar to those of total alkali of aegypti. Norisopoldine inhibits the activation of macrophages and the release of pro-inflammatory factors by down-regulating the activity of the MAPKs signaling pathway, thereby significantly reducing the release of nitric oxide, tumor necrosis factor, and interleukin-1β from RAW264.7 macrophages.
However, pharmacokinetic studies show that the bioavailability of norisoboldine is only 2.7%, and the blood concentration of the metabolite norisoboldine-9-O-α-glucuronide is greatly It is higher than its prototype drug, and its residence time in the body is also longer than that of the prototype drug. Animal experiments of norisopoldine have shown that it can significantly reduce the severity of arthritis in mice (CIA), protect joints from damage, and reduce the content of anti-collagen antibodies in serum. The pharmacological effects are clear and the toxic side effects are small. However, Because it is slightly soluble in water, its rapid metabolism in the body and low bioavailability in blood concentration greatly limit its clinical application.
Main reference materials
[1] CN201110420168.4 A method for separation and purification of norisoboldine monomer
[2] Study on the pharmacokinetics and bioavailability of norisoboldine and its metabolites
[3] CN201510469672.1 Norisoboldine chitosan microspheres and preparation method thereof
