CN112494598B - 一种用于治疗咽炎的有效部位组合物及其应用 - Google Patents
一种用于治疗咽炎的有效部位组合物及其应用 Download PDFInfo
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Abstract
本发明公开了一种用于治疗咽炎的中药有效部位组合物，其包括总黄酮提取物和总皂苷提取物；所述的总黄酮提取物提取自野菊花、橘络与红姑娘，所述的总皂苷提取物提取自沿阶草。按照重量份数计，其包括总黄酮提取物2～8份和总皂苷提取物2～10份。通过实验证明，本发明的有效部位组合物在治疗咽炎方面具有显著的协同效应。本发明产品成分明确，疗效稳定，服用量小，安全性高，不仅符合传统中医药治疗慢性咽炎的组方原则，而且符合中药现代化的发展要求，对推动医药产业从仿制向自主创新转变以及对推动我国中药国际化具有重要意义。
Description
技术领域
本发明属于中药领域，具体涉及一种用于治疗咽炎的有效部位组合物及其应用。
背景技术
慢性咽炎为咽黏膜、黏膜下及淋巴组织的慢性炎症。弥漫性咽部炎症常为上呼吸道慢性炎症的一部分；局限性咽部炎症则多为咽淋巴组织炎症。本病在临床中常见，病程长，症状容易反复发作。慢性咽炎多见于成年人，儿童也可出现。全身症状均不明显，以局部症状为主。各型慢性咽炎症状大致相似且多种多样，如咽部不适感、异物感、咽部分泌物不易咯出、咽部痒感、烧灼感、干燥感或刺激感，还可有微痛感。由于咽后壁通常因咽部慢性炎症造成较黏稠分泌物粘附，以及由于鼻、鼻窦、鼻咽部病变造成夜间张口呼吸，常在晨起时出现刺激性咳嗽及恶心。由于咽部异物感可表现为频繁吞咽。咽部分泌物少且不易咳出者常表现为习惯性的干咳及清嗓子咳痰动作，若用力咳嗽或清嗓子可引起咽部黏膜出血，造成分泌物中带血。
西医临床体征检查，咽后壁或舌根淋巴滤泡增生，重者有角化物附着，表现为咽喉部有异物感，但进食正常。诊断依据，咽部或喉部检查未见占位性病变，局部有淋巴滤泡增生，甚至有角化物附着，食道检查正常。目前市面上的西药咽喉片主要是华素片与明治地喹氯胺含片两种，这两种药片都不含抗生素。其中，华素片的主要成分是西地碘，碘分子具有超强杀菌抗感染作用，可迅速杀灭口腔、咽喉部位各种致病微生物，包括细菌繁殖体、真菌、芽孢、病毒。华素片独特的口含消炎方式直接作用于病症感染部位，清凉薄荷滋润咽喉，可有效治疗咽喉疾病，但长期服用碘和碘化合物可能会产生精神抑郁、神经过敏等不良反应。另外，明治地喹氯胺含片的主要成分是地喹氯胺，它通过破坏细菌表面从而杀菌。西药物治疗见效快，治疗短期内效果明显，但对身体有一定影响、部分人群服药后会导致一些副作用；西医治疗疾病采用对症治疗为目的，消除症状效果明显，治疗针对病症特征，但在治疗慢性咽炎时易反弹复发。
中医认为慢性咽炎与情志郁结，气机不利有关。如同梅核塞于咽喉，咯之不出，咽之不下，故名为“梅核气”。多由素体肺肾阴虚，或风热喉痹反复发作，余邪留滞不清，伤津耗液，使阴液亏损，咽喉失于濡养，兼之虚火上的，从而导致本病的发生。本病多在脏腑阴阳气血虚损的基础上发生，一般病程较长。临床所见，以阴虚为多，阳虚相对少见，亦有在阴虚或阳虚的基础上兼挟"痰凝"或"瘀血"而表现为虚中挟实者，辩证时须仔细区分。慢性咽炎以虚证居多，治疗大法以补为主，或滋阴降火，或温补脾肾、引人归原。因病程较长，难取速效，故长期服药，滋阴须防其腻，温补宜防其燥。至于挟痰挟瘀者，因其本乃虚中挟实，不可一味攻伐伤其正气，致病情更为缠绵。除药物治疗外，适当的心理疏导，使心情舒畅，气机调达，亦十分关键。
但是由于中药饮片有效成分含量低，服用量大，不方便，药味苦，所以不易被患者接受，但经过现代科学工艺的提取，使得有效成分含量提高，用量减少，服用方便，而且良药不再苦口，增加了接受的人群。
发明内容
本发明针对上述现有技术存在的不足，提供一种用于治疗咽炎的有效部位组合物，对于咽炎的治疗具有显著疗效，可应用于各种制剂。
具体技术方案如下：
本发明的目的之一是提供一种用于治疗咽炎的中药有效部位组合物，其特征在于，包括总黄酮提取物和总皂苷提取物；
所述的总黄酮提取物提取自野菊花、橘络与红姑娘(Physalis peruviana L.)，所述的总皂苷提取物提取自沿阶草。
本发明中所述的红姑娘为茄科植物酸浆果(Physalis peruviana L.)的果实的干制品。
实验证实，本发明的中药有效部位组合物具有显著的协同效应，对于咽炎疗效显著。
进一步，所述的中药有效部位组合物中，按照重量份数计，包括总黄酮提取物2～8份和总皂苷提取物2～10份。
再进一步，所述的中药有效部位组合物中，按照重量份数计，包括总黄酮提取物6份和总皂苷提取物5份。
进一步，所述的总黄酮提取物中总黄酮的含量不低于50wt％，所述的总皂苷提取物中的总皂苷的含量不低于50wt％。
进一步，上述中药有效部位组合物的制备方法包括如下步骤：
(1)总黄酮提取物制备：取药材野菊花、橘络和红姑娘为原料，加以其总重量计15～30倍量的80％～95％乙醇，70～80℃回流提取1～3次，每次2h，滤过，滤液合并，减压回收乙醇，加水稀释浓缩液，以0.8～1mL/min的流速通过大孔树脂柱，先以6～10倍柱体积去离子水洗脱，再以8～15倍柱体积70％乙醇洗脱，收集洗脱液，减压回收乙醇并浓缩，干燥，得总黄酮提取物；
(2)总皂苷提取物制备：取药材沿阶草为原料，加入以其重量计8～10倍量的75％～85％乙醇75～80℃回流提取1～1.5h，过滤，得滤液，滤渣重复操作提取一次，合并两份滤液，减压回收乙醇至相对密度为1:1～1:2，加入水饱和正丁醇萃取三次，合并正丁醇部分，回收正丁醇至每mL含1～1.5g生药，以0.8～1mL/min的流速通过大孔树脂柱，先以4～5倍柱体积的蒸馏水洗脱，再以4～5倍柱体积的70％乙醇洗脱，收集洗脱液，减压回收，干燥得总皂苷提取物。
再进一步，步骤(1)中，野菊花、橘络和红姑娘的质量比为1：(1～3)：(1～4)，最优选为1：2：3。
再进一步，步骤(1)中使用的大孔树脂柱为HPD-100大孔树脂柱，步骤(2)中使用的大孔树脂柱为D101大孔树脂柱。
本发明中所述的乙醇的百分比均为体积百分比。
本发明的目的之二是提供上述中药有效部位组合物在制备治疗咽炎的药物中的应用。
按照各有效部位的重量配比取上述两份有效部位提取物，不加辅料或加入药剂学常规辅料，制成药学上可以接受的剂型，片剂、颗粒剂、硬胶囊剂、丸剂、滴丸、或软胶囊等。
以上所说的药剂学常规辅料包括但不限于粘合剂，如纤维素衍生物、藻酸盐、明胶和聚乙烯吡咯烷酮；稀释剂，淀粉、糖粉、糊精、乳糖、微晶纤维素、甘露醇；润滑剂，如滑石粉、硬脂酸镁、微粉硅胶和聚乙二醇；崩解剂，如羧甲淀粉钠、低取代羟丙甲纤维素、交联羧甲基纤维素；吸收促进剂，如季铵化合物；表面活性剂，如十六烷醉、十二烷基硫酸钠；赋形剂，如乳糖、淀粉、蔗糖、糊精、碳酸钙、硫酸铝、氧化镁、硬脂酸镁，碳酸氢钠、甘油、丙二醇、聚乙二醇、山梨糖醇、羊毛脂、凡士林、微晶纤维素、蜂腊、木腊、液体石蜡、树脂、高级蜡、压敏胶、半合成脂肪酸脂等。
本发明的有益效果如下：
本发明以传统中医理论为指导思想，同时结合国内外的相关研究成果，在治疗慢性胃炎的方面效果显著。方中以野菊花为君药，野菊花味苦、辛、凉，具有清热解毒，疏风散热，散瘀，明目，降血压之功效，用于治疗咽炎、肺炎、白喉、胃肠炎、疔、痈等症，故为君。红姑娘为臣药，具有清热，解毒，利尿之功效，用于治热咳，咽痛，痢疾，水肿，疔，丹毒。红姑娘与野菊花配伍具有清热降火解读之功效，故为臣药。沿阶草为佐药，具有滋阴润肺；益胃生津；清心除烦之功效，用于肺燥干咳、肺痈、阴虚劳嗽、津伤口渴、咽喉疼痛、肠燥便秘等症，故为佐药。橘络为使药，橘络味苦、甘，性平。脾、肺经。通络，化痰止咳。用于咳嗽痰多，胸胁作痛。黄芪性平，能够调和诸药，引药直达病所，故为使药。本方中各药物相辅相成，缺一不可，按中医君、臣、佐、使理论合理组方，共奏清热解毒、疏风散热、清心除烦、滋阴降火，润燥利咽之功。
通过实验证明，本发明的有效部位组合物在治疗咽炎方面具有显著的协同效应。
本发明产品成分明确，疗效稳定，服用量小，安全性高，不仅符合传统中医药治疗慢性咽炎的组方原则，而且符合中药现代化的发展要求，对推动医药产业从仿制向自主创新转变以及对推动我国中药国际化具有重要意义。经查阅相关文献和专利，未检索到以野菊花、橘络、沿阶草、红姑娘的有效部位作为原料，生产加工并制成适宜药物制剂用于慢性咽炎的治疗或辅助治疗的报道。
具体实施方式
以下结合实例对本发明的原理和特征进行描述，所举实例只用于解释本发明，并非用于限定本发明的范围。实际生产中的用量不仅限于实例中的药物用量，可以用吨、千克、克等计量单位，但是各原料药物的用量配比仍依据本方明的技术方案。
具体实施方式中，总黄酮含量的测定方法为：精密称取在120℃减压干燥至恒重的芦丁对照品约10mg，置100mL容量瓶中，加70％乙醇70mL，置水浴微热使溶解，放冷，加70％乙醇至刻度，摇匀，即得。精密吸取对照品溶液1.0、2.0、3.0、4.0、5.0mL，分别置于10mL量瓶中，分别加水4.0、3.0、2.0、1.0、0，加5％亚硝酸钠溶液0.3mL，摇匀，放置6min，加10％硝酸铝溶液0.3mL，摇匀，放置6min，加4％氢氧化钠4mL，再加水刻度，摇匀，放置12min；同法制得空白溶液。于510nm处测定吸光度。以吸收度为纵坐标，浓度为横坐标，绘制标准曲线。取供试品100mg，至100ml容量瓶中，加70％乙醇至刻度，摇匀、滤过。精密量取滤液3mL，加2mL水，加5％亚硝酸钠溶液0.3mL，摇匀，放置6min，加10％硝酸铝溶液0.3mL，摇匀，放置6min，加4％氢氧化钠4mL，再加水刻度，摇匀，放置12min。于510nm测定吸光度，并根据标准曲线方程计算含量。
具体实施方式中，总皂苷含量的测定方法为：精密称取齐墩果酸对照品5.00mg,加甲醇溶解定容至25mL,制成每mL含齐墩果酸对照品0.2mg的溶液，即得。精密移取对照品溶液0，0.1,0.2,0.3,0.4,0.5,0.6mL分别加入7支干燥具塞试管中,水浴挥干溶剂,各加入新鲜配制的50mL/L香草醛冰醋酸溶液0.2mL,高氯酸0.8mL,于60℃恒温水浴加热15min,取出,流水冷却2min,加冰醋酸5ml,摇匀,随行试剂空白,照分光光度法试验,在544nm波长处测定吸收度。以吸收度为纵坐标,齐墩果酸量为横坐标,绘制标准曲线。取木通黄芪总皂苷有效部位10mg，加甲醇溶解定容至10mL。取供试品溶液0.5mL，置干燥具塞试管中,水浴挥干溶剂,各加入新鲜配制的50mL/L香草醛冰醋酸溶液0.2mL,高氯酸0.8mL,于60℃恒温水浴加热15min,取出,流水冷却2min,加冰醋酸5mL,摇匀，测定吸光度，代入标准曲线中，计算即得。
实施例1
制备治疗咽炎的胶囊，处方如下：总黄酮提取物120g、总皂苷提取物100g。
总黄酮提取物的制备方法如下：取以重量份数计的野菊花1份、橘络2份和红姑娘3份为原料，加以其重量计20倍量90％乙醇，75℃回流提取3次，每次2h，滤过，将滤液合并，减压回收乙醇，加水稀释浓缩液至原滤液体积，以1mL/min的流速通过HPD-100大孔树脂柱，先以8倍柱体积去离子水洗脱，再以10倍柱体积70％乙醇洗脱，收集洗脱液，减压回收乙醇并浓缩，干燥，得总黄酮提取物，测得其中的总黄酮含量为52.3％。
总皂苷提取物制备：取药材沿阶草为原料，加入以其重量计10倍量的80％乙醇80℃回流提取1.5h，过滤，得滤液，滤渣重复操作提取一次，合并两份滤液，减压回收乙醇至相对密度为1:1.5，加入等质量水饱和正丁醇萃取三次，合并正丁醇部分，回收正丁醇至每mL含1～1.5g生药，以1mL/min的流速通过D101大孔树脂，先以5倍柱体积的蒸馏水洗脱，再以5倍柱体积的70％乙醇洗脱，收集洗脱液，减压回收，干燥得总皂苷提取物，测得其中总皂苷含量为56.1％。
制法：按照处方取上述各有效部位提取物，粉碎，混合，加入淀粉至300g，混匀，制粒，将所得装入1号胶囊，每粒胶囊含内容物0.3g，即得。
实施例2
制备治疗咽炎的片剂，处方如下：总黄酮提取物120g、总皂苷提取物100g。
总黄酮提取物、总皂苷提取物的制备方法同实施例1。
制法：按照处方取上述各有效部位提取物，粉碎，混合，加入羟丙甲纤维素40g、微晶纤维素20g、微粉硅胶5g再加乳糖至300g，混匀，制粒，压制成片，即得。
实施例3
制备治疗咽炎的软胶囊，处方如下：总黄酮提取物140g、总皂苷提取物100g。
总黄酮提取物、总皂苷提取物的制备方法同实施例1。
制法：按照处方取上述各有效部位提取物，加聚乙二醇400至总量300g，混匀，压制软胶囊，即得。
实施例4
制备治疗咽炎的颗粒，处方如下：总黄酮提取物240g、总皂苷提取物200g。
黄酮提取物、总皂苷提取物的制备方法同实施例1。
制法：按照处方取上述各有效部位提取物，加10％淀粉，90％乙醇制粒，即得。
实施例5
制备治疗咽炎的颗粒，处方如下：总黄酮提取物3kg、总皂苷提取物2kg。
黄酮提取物、总皂苷提取物的制备方法同实施例1。
制法：按照处方取上述各有效部位提取物，加10％淀粉，90％乙醇制粒，每袋封装5g制得的颗粒，即得。
取实施例5所制颗粒剂治疗140例咽炎患者。治疗方案：口服，一次1袋，一日2次，8周为一个疗程。结果：治愈78例，好转41例，显效13例，无效8例，总有效率为94.3％。
对比例1
取沿阶草100g、红姑娘60g、橘络40g、野菊花20g，加10倍质量水煎煮2h，滤过，药渣再加8倍量水煎煮1.5h，滤过，合并滤液，减压浓缩，减压干燥，粉碎，加入淀粉至300g，装入1号胶囊，每粒内容物0.3g，制成胶囊剂。
对比例2
取实施例1制得的总黄酮提取物20g、实施例1制得的总皂苷提取物200g，加入淀粉至300g，装入1号胶囊，每粒内容物0.3g，制成胶囊剂。
对比例3
取实施例1制得的总黄酮提取物220g，加淀粉至300g，装入1号胶囊，每粒内容物0.3g，制成胶囊剂。
对比例4
取实施例1制得的总皂苷提取物220g，加淀粉至300g，装入1号胶囊，每粒内容物0.3g，制成胶囊剂。
对比例5
制备治疗咽炎的胶囊，处方如下：总黄酮提取物120g、总皂苷提取物100g。
与实施例1的区别在于，对比例5的总黄酮提取物提取自质量比为1:2的野菊花与橘络的混合物，提取方法同实施例1；总皂苷提取物同实施例1。
将上述处方药物共220g，加淀粉至300g，装入1号胶囊，每粒内容物0.3g，制成胶囊剂。
实验
为进一步验证本发明的药理作用，用本发明的胶囊进行动物药效学实验。
验证其对大鼠咽炎的影响。
1试剂、样品和动物
1.1试剂
二甲苯，天津市化学试剂三厂；乙醚(分析纯)，天律化学试剂三厂；0.9％氯化生理盐水，天津市风船化学试剂科技有限公司；角叉菜胶，SIDMA公司，羧甲基纤维素钠，北京风礼精求商贸有限责任公司；注射用氨苄西林钠，山东鲁抗医药股份有限公司。
1.2供试样品
使用实施例1以及对比例1～5获得的胶囊剂作为实验对象。
将实施例1标记为样品1，对比例1～5依次标记为样品2～6。
对照组：慢严舒柠咽炎片，国药准字Z61020304西安科力药业有限公司
给药前，取上述相应制剂，取胶囊内容物与慢严舒柠咽炎片，研细，采用纯水配制成所需要的药物浓度2.6g/ml的溶液，按人鼠给药剂量体表面积折算公式计算出小鼠给药剂量为灌胃39.05g·kg-1·d-1，大鼠给药剂量为灌胃48.28g·kg-1·d-1。
1.3实验动物
昆明种小鼠(20±2)g，雌雄各半，SPF级，由山东中医药大学试验动物中心提供。
SD大鼠(200±20)g，雌雄各半，由山东中医药大学试验动物中心提供。
2实验方法
2.1二甲苯致小鼠耳肿胀实验
2.1.1动物分组：取小鼠80只，雌雄各半，随机分为8组，每组10只，即空白组、对照组、样品1组、样品2组、样品3组、样品4组、样品5组、样品6组。每日灌胃一次，连续给药3天。
2.1.2实验方法：在末次给药1h后，小鼠右耳涂以二甲苯试剂50ul/只，左耳不涂作自身空白对照。涂抹二甲苯30min后，将小鼠脱颈椎处死，用内径8mm的打孔器打下左、右双耳相应部位的对等圆形耳片。在屯子天平上称重，右耳片质量减去左耳片质量之差为肿胀度，比较各组的肿胀度，并计算肿胀抑制率：肿胀抑制率＝(阴性对照组肿胀度-用药组肿胀度)/阴性对照组肿胀度。实验结果见表1。
表1对二甲苯致小鼠耳廓肿胀度的影响
2.2棉球所致大鼠肉芽肿增生实验
2.2.1动物分组：取SD大鼠，随机分为8组，每组10只，即空白组、对照组、样品1组、样品2组、样品3组、样品4组、样品5组、样品6组。每日灌胃一次，连续给药10天。
2.2.2实验方法：SD大鼠在乙醚浅麻醉及无菌坏境下作腹部小切口，用血管钳和镊子扩充大鼠皮下组织，在双侧腹股沟皮下植入30±1mg的灭菌棉球各1个，然后缝合。用前棉球加氨苄西林钠1mg/0.1mL，60℃烘箱中烤干，以防感染。除空白对照组(生理盐水)外，其余各给药组在手术当天开始灌胃给药，1次/d，连续10d。在第10天给药1h后股动脉放血处死大鼠，剥离取出棉球肉芽织织，剔除周削多余的脂肪，于70℃烘箱中干燥5h后取出，称重，将称地得的质量减去棉球重量，即为肉芽肿的质量，计算肉芽抑制率。肉芽抑制率＝(空白对照组肉芽质量-治疗组肉芽质量)/空白组肉芽质量。实验结果见表2。
表2对大鼠棉球肉芽肿形成实验的影响
2.3角叉菜胶致大鼠足肿胀实验
2.3.1动物分组：取SD大鼠，随机分为8组，每组10只，即空白组、对照组、样品1组、样品2组、样品3组、样品4组、样品5组、样品6组。每日灌胃一次，连续给药5天。
2.3.2实验方法：末次给药30min后，在每鼠右后足跖部皮下注入1％的角叉菜胶0.1mL/只致炎，于致炎前后不同时间点测量踝关节周长，每隔一小时测量一次，共测四次，计算肿胀度(致炎后踝关节周长-致炎前踝关节周长)。实验结果见表3。
表3对角叉菜胶所致的组肿胀的影响
2.4羧甲基纤维素钠致小鼠腹腔白细胞游走实验
2.4.1动物分组：取小鼠80只，雌雄各半，随机分为8组，每组10只，即空白组、对照组、样品1组、样品2组、样品3组、样品4组、样品5组、样品6组。每日灌胃一次，连续给药3天。
2.4.2试验方法：末次给药30min后，除对照组外向各组小鼠腹腔注射1％羧甲基纤维素钠0.5ml，30min后处死动物，向每只小鼠腹腔注射0.9％生理盐水5ml，轻轻揉按腹部，抽取腹腔液滴于细胞板上，常规于光镜下进行白细胞计数。实验结果见表4。
表4对羧甲基纤维素钠致小鼠腹腔白细胞游走实验的影响
通过以上二甲苯致小鼠耳肿胀实验、棉球所致大鼠肉芽肿增生实验、角叉菜胶致大鼠足肿胀实验、羧甲基纤维素钠致小鼠腹腔白细胞游走实验，样品1组不论是在抑制肿胀和肉芽肿增生方面，还是在缓解足肿胀和白细胞游走实验，都远远优于其他样品组，说明所选择的有效部位组合物效果显著，产生了显著的协同效应。
以上所述仅为本发明的较佳实施例，并不用以限制本发明，凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
Claims (4)
1.一种用于治疗咽炎的中药有效部位组合物，其特征在于，包括总黄酮提取物和总皂苷提取物；
所述的总黄酮提取物提取自野菊花、橘络与红姑娘，所述的总皂苷提取物提取自沿阶草；
按照重量份数计，包括总黄酮提取物 6份和总皂苷提取物5份；
其制备方法包括如下步骤：
（1）总黄酮提取物制备：取药材野菊花、橘络和红姑娘为原料，加以其重量计15～30倍量80%～95%乙醇，70～80℃回流提取1～3次，每次2h，滤过，滤液合并，减压回收乙醇，加水稀释浓缩液，以0.8～1mL/min的流速通过大孔树脂柱，先以6～10倍柱体积去离子水洗脱，再以8～15倍柱体积70%乙醇洗脱，收集洗脱液，减压回收乙醇并浓缩，干燥，得总黄酮提取物；
（2）总皂苷提取物制备：取药材沿阶草为原料，加入以其重量计8～10倍量的75%～85%乙醇75～80℃回流提取1～1.5h，过滤，得滤液，滤渣重复操作提取一次，合并两份滤液，减压回收乙醇至相对密度为1:1～1:2，加入水饱和正丁醇萃取三次，合并正丁醇部分，回收正丁醇至每mL含1～1.5g生药，以0.8～1mL/min的流速通过大孔树脂柱，先以4～5倍柱体积的蒸馏水洗脱，再以4～5倍柱体积的70%乙醇洗脱，收集洗脱液，减压回收，干燥得总皂苷提取物；
步骤（1）中，野菊花、橘络和红姑娘的质量比为1：2：3。
2.根据权利要求1所述的中药有效部位组合物，其特征在于，所述的总黄酮提取物中总黄酮的含量不低于50wt%，所述的总皂苷提取物中的总皂苷的含量不低于50wt%。
3.一种如权利要求1或2所述的中药有效部位组合物在制备治疗咽炎的药物中的应用。
4.根据权利要求3所述的应用，其特征在于，以所述的中药有效部位组合物为有效成分，制备成片剂、颗粒剂、硬胶囊剂、丸剂或软胶囊。
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