CN105726802A - 一种八正片的制备方法及应用 - Google Patents
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Abstract
本发明属于中药制剂领域,具体提供一种八正片的制备方法,由栀子10g、车前子10g、瞿麦10g、萹蓄10g、滑石10g、大黄10g、川木通10g、灯芯草5g、甘草10g作为原料药制成,按比例取所述中药材,加水煎煮提取,收集提取液,滤过,将滤液减压浓缩至相对密度为1.02~1.18的浓缩液,上HPD400大孔吸附树脂柱,用水溶液洗脱,收集洗脱液,浓缩成浸膏或浓缩液,喷雾干燥,制粒,压片,每片重0.6g,使得载药量有很大提高,本发明还提供了八正片在制备抑制人血管内皮EC?304细胞增殖药物中的应用。
Description
技术领域
本发明涉及中药制剂技术领域,具体涉及一种八正片的制备方法及应用。
背景技术
八正片记载于中药新药转正标准,处方为栀子10g、车前子10g、瞿麦10g、萹蓄10g、滑石10g、大黄10g、川木通10g、灯芯草5g、甘草10g,以上八味,采用普通工艺制备成片剂。清热、利尿,通淋。用于湿热下注,小便短赤,淋漓涩痛,口燥咽干。广谱抗菌,速效利尿,消炎止痛,促进愈合。综合调理,提高免疫,减少复方,纯天然中药制剂,副作用小,减少耐药。口服,一次3-4片,一日3次,每片0.6g。
现有技术中,尚未有八正片在提取制备方面采用大孔树脂技术的报道,而采用普通方法,工艺粗糙、复杂、落后,杂质多,导致患者用量过大,不方便服用,严重影响了本品在临床上应用。
发明内容
发明目的:为了解决上述问题,本发明的目的在于提供一种八正片的制备方法。
本发明的另一个目的在于提供一种八正片在制备抑制人血管内皮EC-304细胞增殖药物中的应用。
技术方案:本发明的目的是通过如下的方案实现的:
一种八正片的制备方法,由栀子10g、车前子10g、瞿麦10g、萹蓄10g、滑石10g、大黄10g、川木通10g、灯芯草5g、甘草10g作为原料药制成,所述的方法由下列步骤组成:取上述中药材,加水煎煮提取,收集提取液,滤过,将滤液减压浓缩至相对密度为1.02~1.18的浓缩液,上HPD400大孔吸附树脂柱,用水溶液洗脱,收集洗脱液,浓缩成浸膏或浓缩液,喷雾干燥,制粒,压片,每片重0.6g。
所述八正片的制备方法,制备方法中煎煮条件为:第一次加水为药材重量的8-12倍量,煎煮1-2h,第二次加水为药材重量的6-10倍量,煎煮1-2h。
所述八正片的制备方法,制备方法中喷雾干燥条件为:进风温度为100-120℃,出风温度为80-90℃,物料温度为70-90℃,雾化压力为0.2-0.4兆帕,喷雾速度为5-10ml/s。
所述八正片在制备抑制人血管内皮EC-304细胞增殖药物中的应用,八正片由栀子10g、车前子10g、瞿麦10g、萹蓄10g、滑石10g、大黄10g、川木通10g、灯芯草5g、甘草10g作为原料药制成,制备方法由下列步骤组成:取上述中药材,加水煎煮提取,收集提取液,滤过,将滤液减压浓缩至相对密度为1.02~1.18的浓缩液,上HPD400大孔吸附树脂柱,用水溶液洗脱,收集洗脱液,浓缩成浸膏或浓缩液,喷雾干燥,制粒,压片,每片重0.6g。
所述八正片在制备抑制人血管内皮EC-304细胞增殖药物中的应用,制备方法中煎煮条件为:第一次加水为药材重量的8-12倍量,煎煮1-2h,第二次加水为药材重量的6-10倍量,煎煮1-2h。
所述八正片在制备抑制人血管内皮EC-304细胞增殖药物中的应用,制备方法中喷雾干燥条件为:进风温度为100-120℃,出风温度为80-90℃,物料温度为70-90℃,雾化压力为0.2-0.4兆帕,喷雾速度为5-10ml/s。
现有技术中,八正片口服,一次3-4片,一日3次,每片0.6g,采用本发明制备成的八正片每片0.6g,但含有的药材量比原来多,在同样服用量的情况下具有更多活性成分。
具体实施方式
以下通过实施例形式,对本发明的上述内容再作进一步的详细说明,但不应将此理解为本发明上述主题的范围仅限于以下的实例,凡基于本发明上述内容所实现的技术均属于本发明的范围。
实施例1:取栀子10g、车前子10g、瞿麦10g、萹蓄10g、滑石10g、大黄10g、川木通10g、灯芯草5g、甘草10g,加水煎煮提取,第一次加水为药材重量的8倍量,煎煮2h,第二次加水为药材重量的6倍量,煎煮2h,收集提取液,滤过,将滤液减压浓缩至相对密度为1.02的浓缩液,上HPD400大孔吸附树脂柱,用水溶液洗脱,收集洗脱液,浓缩成浸膏或浓缩液,喷雾干燥,制粒,压片,每片重0.6g,喷雾干燥条件为:进风温度为100℃,出风温度为90℃,物料温度为70℃,雾化压力为0.4兆帕,喷雾速度为5ml/s。。
实施例2:取栀子10g、车前子10g、瞿麦10g、萹蓄10g、滑石10g、大黄10g、川木通10g、灯芯草5g、甘草10g,加水煎煮提取,第一次加水为药材重量的12倍量,煎煮1h,第二次加水为药材重量的10倍量,煎煮1h,收集提取液,滤过,将滤液减压浓缩至相对密度为1.18的浓缩液,上HPD400大孔吸附树脂柱,用水溶液洗脱,收集洗脱液,浓缩成浸膏或浓缩液,喷雾干燥,制粒,压片,每片重0.6g,喷雾干燥条件为:进风温度为120℃,出风温度为80℃,物料温度为90℃,雾化压力为0.2兆帕,喷雾速度为10ml/s。
实施例3:取栀子10g、车前子10g、瞿麦10g、萹蓄10g、滑石10g、大黄10g、川木通10g、灯芯草5g、甘草10g,加水煎煮提取,第一次加水为药材重量的10倍量,煎煮1.5h,第二次加水为药材重量的8倍量,煎煮1.5h,收集提取液,滤过,将滤液减压浓缩至相对密度为1.10的浓缩液,上HPD400大孔吸附树脂柱,用水溶液洗脱,收集洗脱液,浓缩成浸膏或浓缩液,喷雾干燥,制粒,压片,每片重0.6g,喷雾干燥条件为:进风温度为110℃,出风温度为85℃,物料温度为80℃,雾化压力为0.3兆帕,喷雾速度为7.5ml/s。
实施例4:八正片抑制人血管内皮EC-304细胞增殖的实验研究资料
1实验材料
1.1实验用细胞株:人血管内皮EC-304细胞,南京医科大学实验室细胞库,DMEM+10%FBS常规培养。
1.2实验药物:研究药物:本发明八正片:按实施例3方法制备。药液储液:称取100mg八正片内容物,混悬于5ml无水乙醇中,0.2μm滤器过滤,500μl doff管分装,-20℃存储,同时0.2μm滤器过滤无水乙醇以备对照组之用。
1.3实验试剂:DMEM(GIBCO公司Cat.No.12100-061Lot.No.758137);胎牛血清(浙江天杭生物科技有限公司Lot.No.100419);NaHCO3(上海久亿化学试剂有限公司Cat.No.11810-033Lot.No.1088387);Trypsin(AMRESCO公司批号:2010/04);EDTA(AMRESCO公司批号:2009/10);Penicillin G Sodium Salt(AMRESCO公司批号:2010242);Streptomycin Sulfate(AMRESCO公司批号:2010382);无水乙醇(南京化学试剂有限公司批号:080310182);MTT(Biosharp批号:0793);PBS(实验室自配);
1.4实验器材:莱卡倒置显微镜(德国Leica型号:DM1L);可见-紫外光微孔板检测仪(美国MD公司型号:SPECTRA MAX 190);CO2培养箱(FORMA型号:3111);超净台(苏净集团安泰公司制造型号:SW-CJ-ZFD);纯水仪(美国Spring公司型号:S/N 020579);精密移液器(法国吉尔森公司型号:P2);电子天平(德国赛多利斯有限公司型号:BT323S);全自动高压灭菌锅(日本SANYO公司型号:MLS-3020);台式电热鼓风干燥箱(上海精密实验设备公司型号:DHG9123A);冰箱(西门子公司型号:KG18V21TI);液氮罐(CBS型号:2001);低速离心机(上海安亭科学仪器厂型号:KA-1000);0.2μm滤器(MILLIPORE型号:SLGP033RB);10cm培养皿(NEST公司)、96孔培养板(NEST公司);细胞计数板;离心管、移液管、Tips若干。
2实验方法:1)人血管内皮EC-304细胞用DMEM+10%FBS于37℃、5%CO2进行常规培养(10cm培养皿),当细胞生长至对数期时,收集细胞,弃去培养液,PBS轻洗3遍,加入3ml 0.25%胰蛋白酶-0.04%EDTA,37℃消化2min后,向其中加入5ml完全培养基中和反应,吹打细胞后将其转入离心管中,1000rpm离心5min,调整细胞悬液浓度3×104个/ml。2)将细胞种入96孔培养板中,每孔加入细胞悬液180μl,培养板放入细胞培养箱中(37℃,5%CO2)常规培养。3)根据细胞生长情况,一般长至50%-70%,加入八正片溶液,继续培养24h。4)24h后加入20μl MTT溶液(5mg/ml,即0.5%MTT),继续培养4h。5)4h后扣板法倒去上清,用吸水纸轻轻拍干,每孔加入200μl二甲基亚砜,置摇床上低速振荡10min,使结晶物充分溶解。在酶联免疫检测仪490nm处测量各孔的吸光值。6)同时设置本底(不加细胞,只加培养液),对照孔(细胞、相同浓度的药物溶解介质、培养液、MTT、二甲基亚砜),每组设定6个复孔。7)结果以药物对细胞的抑制率表示:细胞增值抑制率(%)=(对照孔OD值-给药孔OD值)/对照孔OD值×100%。实验重复3次。
3实验结果:MTT法实验后统计结果显示,与对照组比较,当剂量达到5mg/ml时,对人血管内皮EC-304细胞增殖抑制有差异(P<0.05),剂量在10mg/ml时该差异具有显著性(P<0.01),当剂量达到15-20mg/ml时有极显著性差异(P<0.001)。
表1八正片对人血管内皮EC-304细胞增殖抑制影响研究
注:与对照组比较,*P<0.01;**P<0.001
4实验结论:八正片可以抑制人血管内皮EC-304细胞增殖,减少人血管内皮EC-304细胞的细胞生长数目,该作用呈剂量依赖性。
Claims (6)
1.一种八正片的制备方法,由栀子10g、车前子10g、瞿麦10g、萹蓄10g、滑石10g、大黄10g、川木通10g、灯芯草5g、甘草10g作为原料药制成,其特征在于所述的方法由下列步骤组成:取上述中药材,加水煎煮提取,收集提取液,滤过,将滤液减压浓缩至相对密度为1.02~1.18的浓缩液,上HPD400大孔吸附树脂柱,用水溶液洗脱,收集洗脱液,浓缩成浸膏或浓缩液,喷雾干燥,制粒,压片,每片重0.6g。
2.根据权利要求1所述八正片的制备方法,其特征在于制备方法中煎煮条件为:第一次加水为药材重量的8-12倍量,煎煮1-2h,第二次加水为药材重量的6-10倍量,煎煮1-2h。
3.根据权利要求1所述八正片的制备方法,其特征在于制备方法中喷雾干燥条件为:进风温度为100-120℃,出风温度为80-90℃,物料温度为70-90℃,雾化压力为0.2-0.4兆帕,喷雾速度为5-10ml/s。
4.八正片在制备抑制人血管内皮EC-304细胞增殖药物中的应用,其特征在于八正片由栀子10g、车前子10g、瞿麦10g、萹蓄10g、滑石10g、大黄10g、川木通10g、灯芯草5g、甘草10g作为原料药制成,制备方法由下列步骤组成:取上述中药材,加水煎煮提取,收集提取液,滤过,将滤液减压浓缩至相对密度为1.02~1.18的浓缩液,上HPD400大孔吸附树脂柱,用水溶液洗脱,收集洗脱液,浓缩成浸膏或浓缩液,喷雾干燥,制粒,压片,每片重0.6g。
5.根据权利要求4所述八正片在制备抑制人血管内皮EC-304细胞增殖药物中的应用,其特征在于制备方法中煎煮条件为:第一次加水为药材重量的8-12倍量,煎煮1-2h,第二次加水为药材重量的6-10倍量,煎煮1-2h。
6.根据权利要求4所述八正片在制备抑制人血管内皮EC-304细胞增殖药物中的应用,其特征在于制备方法中喷雾干燥条件为:进风温度为100-120℃,出风温度为80-90℃,物料温度为70-90℃,雾化压力为0.2-0.4兆帕,喷雾速度为5-10ml/s。
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