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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