Matrine Isolation By Temperature-Sensitive Molecularly Imprinted Technology And Pharmacokinetics Study Of Matrine And Its Derivatives

Sophora telekinesis, a species of the Sophora genus in Leguminosae family and mainly distributed in Guangxi, is widely used in Chinese herbal medicine that has historically been used in the treatment of cancer and inflammation. Previous phytochemical studies on this plant revealed that quinolizidine alkaloids, flavonoids, and triterpenoids are present as its major constituent. In the past few years, much more attention was paid to the analysis of the quinolizidine alkaloids, including matrine (MT), oxymatrine (OMT), oxysophocarpine (OSC) and sophocarpine (SC), due to their various pharmacological activities. This paper focusses on the determination and extraction MT in the plant. Moreover, metabolism and pharmacokinetic study of several MT derivatives which had highly reactive for anti-tumor was also investigations.Main conclusions of this work are drawn as follows:(1) A rapid HPLC MS/MS method for determination of four quinolizidine alkaloids has been established, the results showed that MT type alkaloids in radix sophorae tonkinensis is dominated by OMT; followed by MT and OSC, also contains a small amount of SC.(2) It was for the first time to establish a HPLC method for determination total MT in radix sophorae tonkinensis using 50% ethanol as medium, sodium bisulfite as reductant which enables the formation of SC, OSC and OMT in the form of MT. It could be using a reverse phase column and then avoid complex mobile phase, provides a quick and convenient method for the determination of total MT. Results show that total MT increased 0.08～0.29% compared with Pharmacopoeia method and basically the same as four of the total alkaloids determination by LC-MS. Which could provide reference and simple method for quality control of traditional Chinese medicinal and evaluation of the extraction process.(3) It was for the first time to the synthesis of a temperature-sensitive MT-imprinted polymer by free-radical cross-linking copolymerization of methacrylic acid at 60 ℃ in the presence of ethylene glycol dimethacrylate as the cross-linker, N-isopropyl acrylamide as the temperature-responsive monomer and MT as the template molecule. Binding experiments and Scatchard analyses revealed that two classes of binding sites were formed on molecular imprinted polymer (MEP) at 50 ℃. Additionally, the thermoresponsive MIP was tested for its application as a sorbent material in the extraction process for the selective separation of MT from radix Sophorae tonkinensis. The MT dissolution curve (before or after using thermoresponsive MIP) is explained by Weibull and Logistic model, the results proved that using thermoresponsive MIP as a sorbent material in the extraction process can significantly improve the MT dissolution efficiency and increase the maximum saturated dissolution. Results showed that the thermoresponsive MIP displayed different efficiency in clean-up and enrichments using the SPE protocol at different temperatures using water as eluent. This approach provides a new method for the extraction of MT in green process.(4) The rat liver microsomal incubation experiments of MT and Its derivatives shown that only 14-thienyl methylene matrine (TMM) occurrence of biological degradation for matrine while 14-(3-Methylbenzyl) matrine (3MBM),14-(4-Methylbenzyl)matrine (4MBM) and 14-(2-Methylnaphthalene) matrine (28#) were not degraded.(5) A rapid, sensitive and selective HPLC MS/MS method has been developed and validated for the simultaneous determination of TMM and MT in rat plasma, the results showed that oral administration of TMM is able to enhance oral bioavailability of MT and greatly extended the half-life of MT. Combination of rat liver microsomal incubation experiments indicating that TMM biotransformation to MT in vivo due to the 14-position of the double bonds, this provided a new idea for the structure modification and the development of new drugs.(6) Pharmacokinetic studies were performed by 3MBM and 4MBM at the equivalent dose, the results showed that two blood drug concentration characteristics are similar, indicating that they possess the same metabolic process. Indicated that 3MBM is more suitable for activity than 4MBM.(7) Rat liver microsomes incubation experiment, pharmacokinetics and pharmacological studies showed that with the increase of LogP the antitumor activity of MT’derivatives also increased, but the plasma concentration decreased, indicating that the structure modification of MT should pay attention to LogP of compounds, in order to improve the development chance into drugs.