The Study Of Preparation Process And Quality Standards Of New Anti-tuberculosis Drug-Tenaidu Capsule

Radix Ranunculi Ternati is a Chinese herbal medicine for the treatment of tuberculosis. It is the dry tuber of Ranunculus ternatus Thunb., which was newly discovered in the 1950s in Xinyang, Henan Province.Clinical use of Radix Ranunculi Ternati Capsule (ministerial standards: WS3-B-2035-95) combined with chemotherapeutic drugs are used to treat scrofula, tuberculosis, tuberculosis of epididymis, etc, which can significantly shorten the treatment cycle of simple application of chemotherapeutics as well as the advantage of reducing resistance and relapse rate.However, the capsule is made directly from the aqueous extracts supplemented with medicinal powder. Such preparation process is rough and lacks rational quality control standards with the active ingredient not clear.In order to clarify the therapeutic basis of Radix Ranunculi Ternati and enrich the active ingredients, to develop an efficacious and quality controlled anti-tuberculosis new traditional Chinese drug, Our project aimed 4-[formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl]butanoic acid as target composition based on pharmacodynamic screening. We have improved the quality standards of Radix Ranunculi Ternati and optimized the extraction and purification process of the anti-tuberculosis extracts. We have also established quality standards of the extraction and preparation process and developed a new antituberculosis drug-Tenaidu Capsule through secondary development of Capsule Radix Ranunculi Ternati.1. The improvement of the quality standards of Radix Ranunculi Ternati. We have added the content assay method of4-[formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl]butanoic acid (HPLC-DAD method) as target composition (HPLC-DAD method). Through the analysis of target composition content in herb with different collected location and harvest time, we have determined the minimum content of 4-[formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl]butanoic acid in herb was 0.0165%.2. The study of intermediate extraction process of Tenaidu Capsule The yield of 4-[formyl-5-(hydroxymethyl)-1H-pyrrol-l-yl]butanoic acid used as index, the extraction process was screened by combination of single factor experiments and orthogonal design. The optimal extraction process was as follows:The herb was soaked in 70% ethanol for three times. Firstly it was extracted with eight times of solvent for 1.5h, then extracted with six times of solvent for 1 h twice. According to the extraction process, the yield and extraction rate of4-[formyl-5-(hydroxymethyl)-1H-pyrrol-l-yl]butanoic acid were 0.0345% and 93.30% respectively.3. The study of intermediate purification process of Tenaidu Capsule D101 macroporous resin was selected as suitable resin by dynamic adsorption and desorption test. The final purification process was optimized by orthogonal design using the combination of 4-[formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl]butanoic acid yield and intermediate content as composite index.The final purification process was as follows:According to 70% of the saturated adsorption capacity as sample volume, after concentrating to 0.5 g·mL-1with, the extract was loaded to macroporous resin column with diameter-height ratio of 1:6, in the speed of 3 BV·h-1. The column was eluted with water,70% ethanol for 4 BV respectively in the speed of 4 BV·h-1. The elute of 70% ethanol was dried then yielded intermediate.4. The study of intermediate quality standards of Tenaidu Capsule10 batches of intermediates were prepared using the selected process from different batches of herbs. Through the analysis of 4-[formyl-5-(hydroxymethyl)-1H-pyrrol-l-yl]butanoic acid content of the 10 batches, content limitation of 4-[formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl]butanoic acid was intially formulated as 1.25%.5. The study of preparation process of Tenaidu CapsuleTenaidu Capsule intermediates used as raw material, both the type and amount of adjuvant were determined by examination of flowability, hydroscopicity, etc. Tenaidu Capsule was prepared and quality standards of preparation were established. The limitation of 4-[formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl]butanoic acid in capsules was 10.0mg·g-1. Preliminary capsule stability test was carried out.