CN110393754A - 一种具有抗呼吸道合胞病毒活性的中药组合物 - Google Patents
一种具有抗呼吸道合胞病毒活性的中药组合物 Download PDFInfo
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Abstract
本发明公开了一种具有抗呼吸道合胞病毒活性的中药组合物,由以下重量份的组分制备而成:鸡血藤25~45份、连翘15~35份、板蓝根20~45份、川椒3~15份。本发明还公开了所述具有抗呼吸道合胞病毒活性的中药组合物的制备方法。本发明制备中药组合物可以有效抑制呼吸道合胞病毒活性,具有剂量较小,毒性小,使用安全的特点,并且原材料易得,成本较低,具有广阔的市场前景。
Description
技术领域
本发明涉及医药卫生领域,具体是一种具有抗呼吸道合胞病毒活性的中药组合物。
背景技术
呼吸道合胞病毒(respiratory syncytial virus,RSV)为副粘病毒科、肺炎病毒属的负链RNA病毒,其感染遍及全世界,是婴幼儿下呼吸道感染的最主要的病原,也老年人以及免疫缺陷人群下呼吸道感染的主要因素。RSV感染主要是损伤细支气管上皮细胞,导致小气道障碍、过度充气以及肺不张,可引起间质性肺炎及毛细支气管炎。目前也没有针对专门针对RSV感染的有效治疗药物。广谱抗病毒药物利巴韦林是唯一可应用于RSV的小分子化学药物,由于存在作用机制不明、疗效不显著和毒副作用等问题,其在临床上的应用十分有限。
中药是中国乃至世界的医学宝库,中药治疗病毒性感染的疾病已经有几千年的历史。中药由于其直接及间接的抗病毒作用特点,使其在治疗病毒类疾病具有独特优势,其能在抗病毒同时兼有解热、抗炎等作用,还能增强机体免疫功能,阻止病毒进入细胞组织,同时很少伤害正常组织细胞,毒副作用较小,故其临床使用前景广阔。因此,很有必要在现有技术的基础之上,寻找开发一种抗呼吸道合胞病毒作用强、不良反应小的中药组合物。
发明内容
本发明的目的在于提供一种具有抗呼吸道合胞病毒活性的中药组合物,以解决上述背景技术中提出的问题。
为实现上述目的,本发明提供如下技术方案:
一种具有抗呼吸道合胞病毒活性的中药组合物,由以下重量份的组分制备而成:鸡血藤25~45份、连翘15~35份、板蓝根20~45份、川椒3~15份。
进一步,由以下重量份的组分制备而成:鸡血藤30~42份、连翘20~30份、板蓝根25~40份、川椒5~12份。
进一步,由以下重量份的组分制备而成:鸡血藤35份、连翘25份、板蓝根33份、川椒7份。
进一步,所述中药组合物为散剂。
一种具有抗呼吸道合胞病毒活性的中药组合物的制备方法,包括以下步骤:
1)按重量份配比取所述原药材中的鸡血藤、连翘和板蓝根,切片,用蒸馏水洗2次,加水适量,加热沸提3次,每次1小时,合并3次提取液,过滤,减压浓缩至小体积,冷冻干燥得提取物;
2)按重量份配比取所述原药材中的川椒,粉碎后过40~80目筛得粉末;
3)将步骤1)所得提取物和步骤2)所得粉末置入混合机中搅拌,充分混匀,最后经粉碎制成超微粉,得到中药组合物。
进一步,步骤1)中,所述减压浓缩的温度为60℃以下。
进一步,步骤3)中,所述粉碎为采用超微粉碎技术进行粉碎。
所述的具有抗呼吸道合胞病毒活性的中药组合物在制备抗呼吸道合胞病毒活性药物中的用途。
本发明公开的一种具有抗呼吸道合胞病毒活性的中药组合物中各味药药效如下:
鸡血藤:性温,味苦微甘,归肝、心、肾经;色赤入血,质润行散;具有活血舒筋,养血调经的功效;主治风湿痹痛,手足麻木,肢体瘫痪,月经不调,经行不畅,痛经,经闭,白细胞减少症,具有扩血管、抗血小板聚集、促进磷代谢等作用。水煎剂对离体及在体蟾蜍心脏有轻度抑制作用;也能增加壮年兔血细胞作用,升高其血红蛋白;能加强子宫的节律性收缩。有调节血脂、抗动脉粥样硬化病变作用。也可治疗风湿性关节炎、红斑性狼疮、血小板减少、急性泄泻等症。
连翘:性微寒,味苦,归肺、心、小肠经;具有清热解毒,消肿散结的功效。连翘或其挥发油、连翘酚具广谱抗菌作用,对多种革兰氏阳性及阴性细菌均有抑制作用。如金黄色葡萄球菌、肺炎双球菌、溶血性链球菌、志贺氏痢疾杆菌、史氏痢疾杆菌、伤寒杆菌、霍乱弧菌、副伤寒杆菌、变形杆菌、大肠杆菌、星形奴卡氏菌等;对亚洲甲型流感病毒、鼻病毒-17型等有抑制作用。用于流感、肺炎、肠炎、流行性淋巴管炎、胸疫及疮肿等。
板蓝根:性苦寒,归心、胃经;具有清热解毒,凉血利咽的功效。本品对多种细菌有抑制作用,对金黄色葡萄球菌、溶血性链球菌、肺炎双球菌、脑膜炎双球菌、伤寒杆菌、副伤寒杆菌、大肠杆菌、肠炎杆菌、八联球菌等具有一定的抑制作用,用于细菌性感染、败血症、肺炎、咽炎、流行性淋巴管炎、马腺疫、胸疫、猪肺疫、猪喘气病、流感、肠炎、水肿病、血痢、肝炎、黄疽及鱼类出血病、烂鳃病和肠炎等病。
川椒:性辛、温,归脾、胃、肾经;具有除湿散寒、活血通络之效,用于胃腹冷痛、呕吐不能食,可与干姜、党参等同用,如大建中汤,还可用于胃脘寒痛、脊背凉痛、蛔虫引起的上腹疼痛、疥疮、水疱丘疹、阴囊湿疹、阴囊潮湿瘙痒等。
与现有技术相比,本发明的有益效果是:
一种具有抗呼吸道合胞病毒活性的中药组合物,可以有效抑制呼吸道合胞病毒的活性,将传统中药与现代制剂理论相结合,具有剂量较小,毒性小,使用安全的特点,并且原材料易得,成本较低,应用前景广泛。
具体实施方式
下面结合实施例进一步说明本发明的实质性内容,但并不以此限定本发明保护范围。尽管参照较佳实施例对本发明作了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的实质和范围。
实施例1
一种具有抗呼吸道合胞病毒活性的中药组合物,由以下重量份的组分制备而成:鸡血藤25份、连翘15份、板蓝根20份、川椒3份。
该中药散剂的制备方法是:
(1)按上述重量份配比分别称取原药材中的鸡血藤、连翘和板蓝根,切片,用蒸馏水洗2次,加水适量,加热沸提3次,每次1小时,合并3次提取液,过滤,20℃减压浓缩至小体积,冷冻干燥得提取物;
(2)按重量份配比称取所述原药材中的川椒,粉碎后过60目筛得粉末;
(3)将步骤(1)所得提取物和步骤(2)所得粉末置入混合机中搅拌,充分混匀,最后经超微粉碎技术粉碎,制成超微粉,得到中药组合物,备用。
实施例2
一种具有抗呼吸道合胞病毒活性的中药组合物,由以下重量份的组分制备而成:鸡血藤42份、连翘30份、板蓝根40份、川椒12份。
该中药散剂的制备方法是:
(1)按上述重量份配比分别称取原药材中的鸡血藤、连翘和板蓝根,切片,用蒸馏水洗2次,加水适量,加热沸提3次,每次1小时,合并3次提取液,过滤,20℃减压浓缩至小体积,冷冻干燥得提取物;
(2)按重量份配比称取所述原药材中的川椒,粉碎后过60目筛得粉末;
(3)将步骤(1)所得提取物和步骤(2)所得粉末置入混合机中搅拌,充分混匀,最后经超微粉碎技术粉碎,制成超微粉,得到中药组合物,备用。
实施例3
一种具有抗呼吸道合胞病毒活性的中药组合物,由以下重量份的组分制备而成:鸡血藤35份、连翘25份、板蓝根33份、川椒7份。
该中药散剂的制备方法是:
(1)按上述重量份配比分别称取原药材中的鸡血藤、连翘和板蓝根,切片,用蒸馏水洗2次,加水适量,加热沸提3次,每次1小时,合并3次提取液,过滤,20℃减压浓缩至小体积,冷冻干燥得提取物;
(2)按重量份配比称取所述原药材中的川椒,粉碎后过60目筛得粉末;
(3)将步骤(1)所得提取物和步骤(2)所得粉末置入混合机中搅拌,充分混匀,最后经超微粉碎技术粉碎,制成超微粉,得到中药组合物,备用。
实施例4
一种具有抗呼吸道合胞病毒活性的中药组合物,由以下重量份的组分制备而成:鸡血藤45份、连翘35份、板蓝根45份、川椒15份。
该中药散剂的制备方法是:
(1)按上述重量份配比分别称取原药材中的鸡血藤、连翘和板蓝根,切片,用蒸馏水洗2次,加水适量,加热沸提3次,每次1小时,合并3次提取液,过滤,20℃减压浓缩至小体积,冷冻干燥得提取物;
(2)按重量份配比称取所述原药材中的川椒,粉碎后过60目筛得粉末;
(3)将步骤(1)所得提取物和步骤(2)所得粉末置入混合机中搅拌,充分混匀,最后经超微粉碎技术粉碎,制成超微粉,得到中药组合物,备用。
实施例5
一种具有抗呼吸道合胞病毒活性的中药组合物,由以下重量份的组分制备而成:鸡血藤30份、连翘20份、板蓝根25份、川椒5份。
该中药散剂的制备方法是:
(1)按上述重量份配比分别称取原药材中的鸡血藤、连翘和板蓝根,切片,用蒸馏水洗2次,加水适量,加热沸提3次,每次1小时,合并3次提取液,过滤,20℃减压浓缩至小体积,冷冻干燥得提取物;
(2)按重量份配比称取所述原药材中的川椒,粉碎后过60目筛得粉末;
(3)将步骤(1)所得提取物和步骤(2)所得粉末置入混合机中搅拌,充分混匀,最后经超微粉碎技术粉碎,制成超微粉,得到中药组合物,备用。
对比例1
一种具有抗呼吸道合胞病毒活性的中药组合物,由以下重量份的组分制备而成:鸡血藤35份、连翘25份。
该中药散剂的制备方法是:
按上述重量份配比分别称取原药材中的鸡血藤和连翘,切片,用蒸馏水洗2次,加水适量,加热沸提3次,每次1小时,合并3次提取液,过滤,20℃减压浓缩至小体积,冷冻干燥得提取物,置入混合机中搅拌,充分混匀,最后经超微粉碎技术粉碎,制成超微粉,得到中药组合物,备用。
对比例2
一种具有抗呼吸道合胞病毒活性的中药组合物,由以下重量份的组分制备而成:板蓝根33份、川椒7份。
该中药散剂的制备方法是:
(1)按上述重量份配比称取原药材中的板蓝根,切片,用蒸馏水洗2次,加水适量,加热沸提3次,每次1小时,合并3次提取液,过滤,20℃减压浓缩至小体积,冷冻干燥得提取物;
(2)按重量份配比称取所述原药材中的川椒,粉碎后过60目筛得粉末;
(3)将步骤(1)所得提取物和步骤(2)所得粉末置入混合机中搅拌,充分混匀,最后经超微粉碎技术粉碎,制成超微粉,得到中药组合物,备用。
对比例3
一种具有抗呼吸道合胞病毒活性的中药组合物,由以下重量份的组分制备而成:鸡血藤35份、连翘25份、板蓝根33份、川椒7份。
该中药散剂的制备方法是:
按上述重量份配比分别称取原药材中的鸡血藤、连翘、川椒和板蓝根,切片,粉碎后过60目筛得粉末,得到中药组合物,备用。
试验例1
(1)试样配制:将实施例1-5和对比例1-3制备的中药组合物分别溶解于DMEM液(含2%小牛血清)中备用,配制浓度为100μg/mL;利巴韦林注射液(河南辅仁怀庆堂制药有限公司生产,批号171114,国药准字H20044698),规格:100mg/1mL(支)。
(2)细胞株与病毒株:人喉表皮癌细胞Hep-2;呼吸道合胞病毒(RSV)A亚型(Long株)。
(3)病毒滴定测定:将活化好的毒株作10-1~10-9系列稀释,使用常规方法感染致密单层Hep-2细胞,每孔100μL,设8个复孔,同时设正常细胞对照组,34.5℃、5%CO2培养箱培养4d,每天观察细胞病变效应(CPE)情况,CPE程度可分为从″-″~″++++″:以最高稀释度不再出现新病变时为终点,无细胞病变时为″-″,25%以下细胞出现病变为″+″,25%~50%为″++″,50%~75%为″+++″,75%~100%为″++++″。
(4)药物对Hep-2细胞主性测定:将胰酶消化后的Hep-2细胞,以5×103个/孔密度接种到96孔培养板上,37℃、5%CO2箱培养培养24h,吸弃培养液后用含2%小牛血清的DMEM培养基稀释药物,其中,将药物倍比稀释成6个系列浓度,起始浓度为500ug/mL,加入96孔板,每孔100μL,每个稀释度设3个复孔,并设正常细胞对照组,继续37℃、5%CO2箱培养48h,观察并记录药物作用于细胞的最大无毒浓度TCO。
(5)中药组合物抗RSV药效学实验:将100TCID50的RSV A亚型病毒液接种于已长成单层的Hep-2细胞中,每孔50μL,在药物无毒浓度范围内加入含不同浓度的中药组合物,每个剂量组重复3孔,所述中药组合物分别为实施例1-5和对比例1-3中得到的中药组合物,于37℃、5%CO2培养箱继续培养。同时设正常细胞对照组、病毒对照组。每天在倒置显微镜下观察细胞形态,以病毒对照组CPE达到75%,且细胞对照正常时为实验观察终点,记录各孔CPE程度。中药组合物抗RSV药效学实验结果见表1。
表1中药组合物抗RSV药效学实验结果
(6)利巴韦林抗RSV药效学实验:将100TCID50的RSV A亚型病毒液接种于已长成单层的Hep-2细胞中,每孔50μL,在药物无毒浓度范围内加入含不同浓度的利巴韦林的维持液。每个剂量组重复3孔,于37℃、5%CO2培养箱继续培养。每天在倒置显微镜下观察细胞形态,以病毒对照组CPE达到75%,且细胞对照正常时为实验观察终点,记录各孔CPE程度。利巴韦林抗RSV药效学实验结果见表2。
表2利巴韦林抗RSV药效学实验结果
样品ug/mL | 500 | 250 | 125 | 62.5 | 31.25 | 15.625 |
利巴韦林 | - | - | - | - | + | + |
在抗病毒活性实验中,Hep-2细胞对照未出现病变,根据表1和12的结果,可以看出,阳性对照药物利巴韦林在31.25ug/ml以上浓度时有明显抑制RSV的作用。通过比较实施例1-5中药组合物的抗RSV药效学实验结果,可以看出,实施例3中药组合物的抗RSV病毒活性效果最佳,通过比较实施例3、对比例1和对比例2中药组合物的抗RSV药效学实验结果,可以看出,采用实施例3配方中部分药材制备的对比例1和对比例2中药组合物对抗RSV病毒活性效果不如实施例3中药组合物,通过比较实施例3和对比例3中药组合物的抗RSV药效学实验结果,可以看出,按照实施例3所述配方采用常规工艺制备中药组合物,其抗RSV病毒活性效果没有实施例3中药组合物的好。随着实施例3中药组合物浓度的增大,Hep-2细胞病变(指病毒引起的细胞病变:细胞变大,融合,脱落形成空斑)的程度逐渐减弱,当药物浓度达到125ug/mL时,细胞病变被完全抑制,即实施例3中药组合物浓度在体外完全抑制RSV增殖的有效浓度为125ug/mL以上。
试验例2
清洁级wistar大鼠,雌雄各半,周龄4-6周,体质量80-100g,清洁级。随机分为四个组,每组10只,各组大鼠除正常对照组外,其余用乙醚浅麻醉后鼻腔滴注RSV 100 TCID50100uL进行病毒接种,连续病毒滴鼻2d。A组:实施例3中药组合物组(0.8g/kg),B组:利巴韦林组(50mg/kg),C组:正常对照组,D组:感染模型组。A,B,D组大鼠在首次滴鼻感染后24h,开始灌胃给药,共给药3d,正常对照组和感染模型组灌胃同样体积的生理盐水。实验结束后处死,取各组大鼠肺组织,福尔马林固定后进行肺组织病理切片研究,观察病理切片发现肺组织的炎性病灶、各级支气管上皮炎性细胞浸润,或有渗出物、肺大泡、肺泡间隔病变等,提示造模成功。
肺部病变评分标准:肺泡病变分为瘀血、间质炎症和气肿,肺内支气管病变分为上皮变性、腔内渗出和管周炎,根据炎症等由轻到重的程度分别标记为1,2,3,4分,极轻度病变为0.5分,累计所有分数,计算出每组动物的均分,结果如表3所示,可见本发明所述具有抗呼吸道合胞病毒活性的中药组合物能有效改善病毒性肺炎症状,治疗效果良好。
表3动物模型病理评分
组别 | n | 组内动物综合评分 |
A | 10 | 3.38±1.17 |
B | 10 | 2.93±1.06 |
C | 10 | 2.22±1.01 |
D | 10 | 4.41±1.03 |
Claims (8)
1.一种具有抗呼吸道合胞病毒活性的中药组合物,其特征在于,由以下重量份的组分制备而成:鸡血藤25~45份、连翘15~35份、板蓝根20~45份、川椒3~15份。
2.根据权利要求1所述的具有抗呼吸道合胞病毒活性的中药组合物,其特征在于,由以下重量份的组分制备而成:鸡血藤30~42份、连翘20~30份、板蓝根25~40份、川椒5~12份。
3.根据权利要求2所述的具有抗呼吸道合胞病毒活性的中药组合物,其特征在于,由以下重量份的组分制备而成:鸡血藤35份、连翘25份、板蓝根33份、川椒7份。
4.根据权利要求1所述的具有抗呼吸道合胞病毒活性的中药组合物,其特征在于,所述中药组合物为散剂。
5.一种如权利要求1-4任一所述的具有抗呼吸道合胞病毒活性的中药组合物的制备方法,其特征在于,包括以下步骤:
1)按重量份配比取所述原药材中的鸡血藤、连翘和板蓝根,切片,用蒸馏水洗2次,加水适量,加热沸提3次,每次1小时,合并3次提取液,过滤,减压浓缩至小体积,冷冻干燥得提取物;
2)按重量份配比取所述原药材中的川椒,粉碎后过40~80目筛得粉末;
3)将步骤1)所得提取物和步骤2)所得粉末置入混合机中搅拌,充分混匀,最后经粉碎制成超微粉,即得。
6.根据权利要求5所述的具有抗呼吸道合胞病毒活性的中药组合物的制备方法,其特征在于,步骤1)中,所述减压浓缩的温度为60℃以下。
7.根据权利要求5所述的具有抗呼吸道合胞病毒活性的中药组合物的制备方法,其特征在于,步骤3)中,所述粉碎为采用超微粉碎技术进行粉碎。
8.如权利要求1-4任一所述的具有抗呼吸道合胞病毒活性的中药组合物在制备抗呼吸道合胞病毒活性药物中的用途。
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