Application of benzoyl aconitine_Kain Industrial Additives

Background and overview^[1]

Aconitum is a processed product of the root of Aconitum, a plant of the Ranunculaceae family, and Aconitum aconite and Aconite are the dried mother roots of Aconitum. Aconite is the seed root of the Aconitum plant of the Ranunaceae family. It is a commonly used traditional Chinese medicine. It has analgesic, diuretic, cardiotonic and metabolic effects. Aconitum tuberous root (mother root) contains aconitine, hypoconitine, neoaconitine, talamine, racemic desmethyl Hengzhou aconitine, isotaladine, neoconitine, Junggar aconitine Alkali, aconitine, nordelsonine, isdelphinidine, benzoyl aconitine, aconitine, sembucin A, sembucin B, 14-acetylalamine, Aconitine, aconitine, aconitine, aconitine, aconitine, aconitine, aconitine, 3-deoxyaconitine, inert base, hocobucin A and B, uracil, aconitum polysaccharide A, B, C, D.

The alkaloids contained in Aconitine include aconitine, neoconitine, hypoconitine, benzoylconitine, benzoyl neoconitine and benzoyl hypoconitine, dl-nor Aconitine, aconitine A, aconitine B, kaurine, aconidine, norsonine, isodelviteine, methamphetamine, hypaconitine, neolin, Junggar aconitine, Xinjiang oil aconitine, North China aconitine, Huangcao aconitine and so on. The diester alkaloids in Aconitine: aconitine, neo-aconitine, and hypoaconitine are the active ingredients and toxic components in Aconite. They need to be processed and detoxified before use. During the processing process, the acetyl group at position 14 is removed. Remove the corresponding monoester alkaloids to obtain the corresponding monoester alkaloids: benzoylaconitine, benzoylneaconitine, benzoylhypoaconitine, and benzoyldeoxyaconitine to achieve the detoxification effect. The processed aconite is called prepared aconitine, in which benzoyl aconitine is an important active ingredient.

Apply^[2-3]

Pharmacological studies have shown that benzoyl aconitine has significant anti-inflammatory and analgesic effects. In addition, relevant domestic research has found that benzoyl neoaconine can inhibit the proliferation of human leukemia cells K562 cells and induce apoptosis. The apoptosis rate is concentration- and time-dependent. Studies have explored the effects of benzoyl neoaconitine on the proliferation and apoptosis of myeloma cells RPMI8226 (RPMI8226). RPMI8226 cells growing in the logarithmic phase were randomly divided into blank control group and benzoylneaconitine (0.1, 0.5, 1, 10, 100 μmo L) groups. After 24 hours, the optical density of each group was measured with CCK8. (A), the half inhibitory mass concentration (IC50) was calculated based on the A value; according to the measured IC50, the logarithmic phase growth RPMI8226 cells were divided into blank control group, negative control group, benzoyl neoaconitine (4, 6 , 8 μmo L) for each group, acted for 12 to 48 hours, used CCK8 to measure the A value of RPMI8226 cells in each group under different action times, and calculated the proliferation inhibition rate of each group; according to the inhibition rate of RMPI8226 proliferation by benzoyl neoconitine The influence pattern was then set up as a blank control group and a benzoylneaconitine (4, 8 μmo L) group. After 24 and 48 h of action, Annexin V/PI double-staining flow cytometry was used to detect the cell apoptosis rate in each group. Results After 24 hours of displaying the effect, the benzoyl neoaconitine in each group was converted into mass concentration. At a mass concentration of 13 to 13 000 ng/m L (i.e. 1 to 10 μmo L), the A value of each group increased with the increase in drug mass concentration. And decreased, P < 0. 05, and its IC50 mass concentration was 1 040 ng/m L; after acting for 12 to 48 hours, the cell proliferation inhibition rate of each group increased with the increase of the added benzoyl neoaconitine and the action time. The cell apoptosis rate in each group increased with the addition of benzoylneaconitine and the effect after 24 and 48 hours, and P < 0.05 in each group. 05. Conclusion: Benzoyl aconitine can inhibit the proliferation of RPMI8226 and induce apoptosis.

In addition, benzoyl aconitine combined with paeoniflorin can increase the anti-inflammatory and analgesic effects of benzoyl aconitine and paeoniflorin; benzoyl aconitine combined with paeoniflorin can increase the anti-inflammatory and analgesic effects of benzoyl aconitine and paeoniflorin. The pharmacodynamic effects of protamine and paeoniflorin in the treatment of rheumatoid arthritis may be mainly related to inhibiting the production of serum PGE2, regulating serum cytokines (IL-1β, TNF-α and VEGF), and inhibiting the synthesis and secretion of IgG. , regulates the expression of STAT1 and STAT3 in the JAK-STAT signaling pathway, and inhibits abnormal secretion and proliferation of synovial cells.

Preparation^[1]

A method for preparing high-purity benzoyl neoaconitine, which adopts high-speed countercurrent chromatography technology and includes the following steps:

1) Prepare a crude extract containing benzoyl neoaconitine as the injection material. The crude extract is a crude extract of Aconitum (for example, it can be a crude extract of raw aconite, a crude extract of prepared aconite or Aconitum crude extract);

2) Prepare a solvent system that constitutes the stationary phase and the mobile phase. The solvent system consists of component A, component B and component C, where component A is a chlorinated alkane and component B is a fatty alcohol. Component C is water, alkaline solvent or acidic solvent. The volume ratio of component A, component B and component C is (2-5):(0-5):(1-5). The solvent After the system is mixed and allowed to stand for stratification, the upper phase is the stationary phase and the lower phase is the mobile phase. Both the stationary phase and the mobile phase can be used to separate benzoyl neoconitine from the Aconitum crude extract in one separation. Components;

3) Fill the column of the countercurrent chromatograph with the stationary phase;

4) Then rotate the main unit, and then pump the mobile phase into the column, or pump the stationary phase and mobile phase into the column at the same time, and then turn the main unit;

5) Inject the sample through the injection valve, receive the target component according to the detector spectrum, and obtain benzoyl neoaconitine after separation.

Main reference materials

[1] CN200810208203.4 Preparation method of high-purity benzoyl neoaconitine

[2] Effects of benzoylneaconitine on the proliferation and apoptosis of myeloma cells RPMI8226

[3] Study on the pharmacodynamics and mechanism of benzoylaconitine combined with paeoniflorin in the treatment of rheumatoid arthritis