CN113575958A - 提高免疫力功能的保健组合物、胶囊及制备方法 - Google Patents
提高免疫力功能的保健组合物、胶囊及制备方法 Download PDFInfo
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- CN113575958A CN113575958A CN202110761142.XA CN202110761142A CN113575958A CN 113575958 A CN113575958 A CN 113575958A CN 202110761142 A CN202110761142 A CN 202110761142A CN 113575958 A CN113575958 A CN 113575958A
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Abstract
本发明涉及一种提高免疫力功能的保健组合物、胶囊及制备方法,复方保健胶囊内容物包括以下重量百分比计的制备原料:NMN25%~35%,灵芝孢子粉10%~15%,铁皮石斛粉10%~15%,防潮剂、助流剂8%~12%,黏合剂、稀释剂30%~40%,润滑剂2%~3%。与现有技术相比,本发明中胶囊外壳可掩盖NNM原料药的不良气味,避免光线、空气和湿气的接触,提高药物的稳定性和生物利用度;复方保健组合物能够提高小鼠胸腺指数,降低小鼠耳肿胀度,促进淋巴细胞增殖,增加小鼠吞噬指数和NK细胞活性。本发明制得的NMN复方保健胶囊含量均匀,体外溶出行为良好,生产工艺简单,稳定性较好,易于放大生产。
Description
技术领域
本发明涉及保健品相关领域,尤其是涉及一种提高免疫力功能的保健组合物、胶囊及制备方法。
背景技术
免疫力低下严重危害着现代人类健康,对于提高人体自身免疫力的研究也在急切进行当中。
烟酰胺单核苷酸(Nicotinamide Mononucleotide,NMN)是烟酰胺腺嘌呤二核苷酸(Nicotinamide Adenine Dinucleotide,NAD+)的前体物质,NMN可以增加细胞内NAD+水平,而NAD+是细胞能量转化的重要辅酶。可以起到恢复人体DNA细胞损伤以及激活人体DNA细胞的效果,提高人体免疫功能。事实上,NAD+水平下降导致的DNA损伤与免疫功能衰退才是各种疾病的关键诱因。
灵芝为多孔菌科真菌赤芝(Ganoderma Lucidum Karst.)或紫芝(Ganodermasinense Zhao,Xu et Zhang)的干燥子实体,亦是传统名贵中药材,具有补气安神、止咳平喘、增强免疫力、抗癌、降压等功效。铁皮石斛为兰科植物(Dendrobium officinale Kimuraet Migo)的干燥茎,具有“滋阴清热,益胃生津”之功效。常用于治疗热病津伤,口干烦渴,食少干呕,病后虚热不退,阴虚火旺,目暗不明,筋骨痿软等。目前已有研究表明,铁皮石斛具有增强免疫力、抗疲劳、抗氧化等多种保健功能。根据中医学基础理论,通过中药配伍以尽可能发挥药物的最大疗效,使其保健效果更好,故大多数石斛相关产品多是中药配伍复方制剂,即在以石斛为主的基础上可配伍灵芝、西洋参等其他滋补药材。
由于NMN对光线、空气和湿度具有一定的敏感性,随着湿度的增加,NMN的含量下降,含水量增加。这种高敏感性和不稳定的生物特性,使得NMN难以长时间储存,直接服用后有效成分易被分解,难以被人体吸收。因此单靠NMN是无法达到长期的良好吸收和充分发挥其药效,并且目前较大剂量的NMN对人体的副作用尚不明确。
发明内容
本发明的目的就是为了克服上述现有技术存在的缺陷而提供一种提高免疫力功能的保健组合物、胶囊及制备方法,使各组分组成一个整体,产生了协同作用,具有增强免疫力的保健功能。
本发明的目的可以通过以下技术方案来实现:
本技术方案的第一个目的是保护一种提高免疫力功能的保健组合物,所述组合物包括以下质量分:烟酰胺单核苷酸2.5~3.5份,灵芝孢子粉1.0~1.5份,铁皮石斛粉1.0~1.5份。
本技术方案的第二个目的是保护一种具有提高免疫力功能的保健组合物胶囊,所述保健组合物囊包括以下重量百分比计的制备原料:
进一步地,所述保健组合物囊包括以下重量百分比计的制备原料:
进一步地,所述黏合剂、稀释剂均为甘露醇、微晶纤维素、乳糖、淀粉中的一种或多种的组合。
进一步优选地,所述黏合剂、稀释剂均为微晶纤维素。
进一步地,所述防潮剂、助流剂均为微粉硅胶、氢化植物油中的一种或者两者的混合。
进一步优选地,所述防潮剂、助流剂均为微粉硅胶。
进一步地,所述润滑剂为硬脂酸镁、硬脂富马酸钠、单硬脂酸甘油酯中的一种或多种的组合。
进一步优选地,所述润滑剂为硬脂富马酸钠。
本技术方案的第三个目的是保护一种上述保健组合物胶囊的制备方法,包括以下步骤:
(1)将NMN原料药过筛备用,铁皮石斛研磨成细粉,按比例分别称取NMN、灵芝孢子粉、铁皮石斛粉、黏合剂、稀释剂,防潮剂、助流剂,润滑剂。
(2)将NMN原料药、灵芝孢子粉和铁皮石斛粉过筛后与步骤(1)中称取的防潮剂、助流剂按等量递加法混匀,制得混合物料A。
(3)将混合物料A与步骤(1)中称取的黏合剂、稀释剂按等量递加法混匀,过筛,制得混合物料B。
(4)将混合物料B与步骤(1)中称取的润滑剂混合均匀,混合3-5min,制得混合物料C,在胶囊板上进行胶囊填充,制得NMN胶囊成品。
进一步地,步骤(1)中所述NMN原料药经过筛后的粒径D≤0.250mm,灵芝孢子粉和铁皮石斛粉研磨成细粉过筛后的粒径D≤0.30mm。
本技术方案中组合物为NMN原料药、灵芝孢子粉、铁皮石斛粉和药学上可以接受的辅料,将原料药、灵芝孢子粉和铁皮石斛粉过筛后与防潮剂、助流剂按等量递加法混合均匀,过60目筛,再与黏合剂、稀释剂,润滑剂混匀后在0#胶囊板上进行装样填充。过筛处理起混合原料药的作用,从而保证组成的均一性。过筛后的原料药具有一定的黏性,容易团聚,加入防潮剂、助流剂混合过筛可改善内容物的流动性。黏合剂、稀释剂具有可压性,有助于提高药物的稳定性。
与现有技术相比,本发明具有以下技术优势:
1)本发明开发了一种提高免疫力功能的中西结合复方保健胶囊,使各组分组成一个整体,产生了协同作用,具有增强免疫力的保健功能。将NMN、灵芝孢子粉、铁皮石斛粉制备成胶囊以增加药物的稳定性。由于原料药在高湿条件下不稳定,故本品不宜采用湿法制粒工艺。在胶囊的制备工艺中,物料直接混合制粒工艺操作过程简单,节约时间,适合工业化生产。
2)本发明制得的具有提高免疫力功能的保健组合物胶囊解决了NMN稳定性差的问题,制得的颗粒疏松圆整、含量均匀,体外溶出行为良好,生产工艺简单,稳定性较好,易于放大生产。
3)本发明涉及的有效成分NMN的质量百分含量在25%~35%,NMN胶囊不易吸潮,溶出度试验表明NMN的溶出的稳定性良好。本发明使用的辅料价廉易得,能够节约生产成本。
4)本发明提供了一种提高免疫力功能的复方保健胶囊的制备方法,经过该制备方法制备的复方保健胶囊能够有效提高小鼠胸腺指数;降低小鼠耳肿胀度;增加小鼠吞噬指数;促进淋巴细胞增殖;增加NK细胞活性。该复方保健胶囊具有增强免疫力的作用。
附图说明
图1为实施例1-3所制备胶囊中NMN在水中的溶出曲线对比图。
图2为实施例3胶囊中NMN在蒸馏水、0.1mol/L盐酸溶液、pH6.8磷酸盐缓冲溶液以及pH6.0磷酸盐缓冲溶液中的溶出曲线,溶出测定方法为中国药典方法:桨法,900mL溶出介质,75rpm,37℃。
具体实施方式
下面结合附图和具体实施例对本发明进行详细说明。
本技术方案中制备具有提高免疫力功能的保健组合物胶囊的方法,包括以下步骤:
(1)将NMN原料药、灵芝孢子粉和铁皮石斛粉过60目筛,按比例分别称取NMN、灵芝孢子粉、铁皮石斛粉、防潮剂、助流剂、黏合剂、稀释剂、润滑剂。
(2)将NMN原料药、灵芝孢子粉和铁皮石斛粉与步骤(1)中称取的防潮剂、助流剂按等量递加法混匀,过60、80或100目筛,优选为过60目筛,制得混合物料A。
(3)将混料A与步骤(1)中称取的黏合剂、稀释剂按等量递加法混匀,过60、80或100目筛,优选为过60目筛,制得混合物料B。
(4)将混合物料B与步骤(1)中称取的润滑剂混合均匀,混合3-5min,制得混合物料C,在0#胶囊板上进行胶囊填充,制得NMN复方保健胶囊。
本技术方案中胶囊外壳可掩盖NNM原料药的不良气味(酸和略微有点苦涩),避免光线、空气和湿气的接触,提高药物的稳定性和生物利用度。该复方保健组合物能够提高小鼠胸腺指数,降低小鼠耳肿胀度,促进淋巴细胞增殖,增加小鼠吞噬指数和NK细胞活性。本发明制得的NMN复方保健胶囊含量均匀,体外溶出行为良好,生产工艺简单,稳定性较好,易于放大生产。
实施例1
本实施例中的配方如下所示:
本实施例中具有提高免疫力功能的保健组合物胶囊的制备方法,其具体操作步骤如下:
(1)称取处方量过60目筛后的NMN原料药、灵芝孢子粉、铁皮石斛粉、微粉硅胶、微晶纤维素。
(2)将NMN原料药、灵芝孢子粉、铁皮石斛粉与微粉硅胶按等量递加法混合均匀,过60目筛,制得混料A。
(3)将混料A与步骤(1)中称取的微晶纤维素按等量递加法混匀,过60目筛,制得混料B。
(4)将混料B与步骤(1)中称取的硬脂富马酸钠混合3-5min,制得混料C,在0#胶囊板上进行胶囊填充,制得NMN复方保健胶囊成品,进行包装,贮存。
实施例2
本实施例中配方如下所示:
本实施例制备NMN复方保健胶囊的具体操作步骤同实施例1。
实施例3
本实施例中配方如下所示:
本实施例制备NMN复方保健胶囊的具体操作步骤同实施例1。
NMN复方保健胶囊是以NMN、铁皮石斛、灵芝为主要成分,采用苯酚-浓硫酸法测定铁皮石斛粉和灵芝孢子粉中粗多糖的含量。通过紫外-可见分光光度法测定标准曲线为:y=0.0560x-0.0064,r=0.9998。每100mg铁皮石斛粉和灵芝孢子粉中粗多糖(以无水葡萄糖计)含量约为88.61mg。
体外溶出度研究是口服制剂的重要评价指标之一,不仅能模拟口服固体制剂在胃肠道中崩解和溶出情况,还能控制药物制剂的质量,因此建立紫外分光光度法测定体外溶出度。由于该复方制剂中NMN在主要成分中的含量较高,故在考察胶囊的体外溶出度时,以NMN的溶出度为评价指标。
从图1可以看出,实施例1制得的胶囊中NMN在水中的溶出时间在60min左右才溶出完全,溶出较慢,效果不理想,而实施例2的溶出时间过早,在15min就溶出完全,实施例3的胶囊溶出时间在30min左右,符合预期值,实施例3胶囊的溶出效果较好。
通过实施例3胶囊中NMN在不同溶出介质中的溶出曲线对比发现,在不同溶出介质中,本发明所制得的胶囊体外溶出良好,其中在0.1mol/L盐酸的溶出效果较好,因此选用0.1mol/L盐酸作为溶出介质。
实施例4
拆方试验
进行复方拆方研究,有助于阐明中药复方的配伍的组成原理及作用机制。采用分组研究法将复方处方分解为组方1(全方)、组方2、组方3和组方4,其中组方2的药物为灵芝孢子粉和铁皮石斛粉,组方3为NMN和灵芝孢子粉,组方4为NMN和铁皮石斛粉,四组实验均以小鼠耳肿胀为指标,实验方法同实施例5中二硝基氯苯诱导的小鼠耳肿胀实验,结果见下表:
表1:各拆方对小鼠耳肿胀的影响
从表中可以看出,组方2、组方3和组方4对小鼠耳肿胀是有效果的,但与全方比较,仍存在差异,全方具有显著的消肿作用(P<0.05),效果最佳。
实施例5
提高免疫力功能的复方保健胶囊的效果验证试验
以实施例3制备的复方保健胶囊内容物灌胃小鼠,验证本发明对于提高免疫力功能的效果。
试验动物:ICR小鼠,清洁级,雄性,体重17~21g,按照运动能力,体重状况分为4组,每组10只。
剂量分组:100mg/kg,300mg/kg,800mg/kg,并设阴性对照组,经口灌胃,连续30d。
试验步骤:
(1)免疫器官重量与脏器系数测定灌胃给药30d,小鼠断颈处死,75%酒精消毒,无菌解剖,取胸腺,脾脏,肝脏,除去残留在脏器上的血液,秤重。最后用所得数据计算脏器指数。脏器指数(g/10g)=器官质量/体质量。
(2)二硝基氯苯诱导的小鼠耳肿胀实验第24d灌胃给药后1h,将1%二硝基氯苯丙酮橄榄油(丙酮:橄榄油体积比1:1)涂抹在小鼠腹部,涂抹剂量为每只100μL。给药第25天,重复以上步骤。给药第29天,各组小鼠右耳前后面均匀涂抹1%二硝基氯苯丙酮橄榄油,涂抹剂量为每只100μL。各组小鼠左耳以同等方式涂抹橄榄油混合液作为对照。1d后,处死小鼠,剪下两边耳壳。剪取部位应在同一部位。取下耳片形状均为椭圆形。小鼠迟发型变态反应由左右耳重量差表示。
(3)碳粒廓清实验灌胃给药30d,将印度墨汁用生理盐水稀释5倍。注射入小鼠体内小鼠躯干发黑(0.1mL/10g)。在注射后2、10min从眼眶静脉丛取血20μL,加入到2mL 0.1%Na2CO3溶液中,600nm波长测定OD值,并将小鼠处死,取肝脏、脾脏,称重。整理得吞噬指数。
(4)脾淋巴细胞体外增殖作用实验
取(1)所得小鼠脾脏,装于含磷酸缓冲盐溶液的培养皿中,以5mL注射塞碾磨,混匀,200目灭菌双层尼龙筛过滤。合并滤液,1500r/min离心5min,弃上清,弹匀后加少量破红裂解液,以磷酸缓冲盐溶液中和破红,1500r/min再次离心5min,弃上清,以含10%胎牛血清的RPMI1640培养液轻轻吹打重悬沉淀。调整小鼠脾细胞为1.5×109个/mL,100μL/孔种板96孔中。设对照组,复方保健胶囊低、中、高剂量组;各组又分单独处理组和联合ConA(5μg/mL)组;每孔加入供试品100μL,于37℃,5%CO2培养箱中培养44h后,每孔加入噻唑蓝(5mg/mL)20μL继续培养4h。以移液枪小心吸去上清,每孔加入150μL二甲基亚砜,37℃孵育20min,490nm测定OD值。计算增殖指数:增殖指数=(OD样品-OD空白)/(OD对照-OD空白)。
(5)小鼠NK细胞的活性测定
靶细胞的传代(YAC-1细胞):实验前24h将靶细胞进行传代培养。应用前以磷酸缓冲盐溶液洗3次,用RPMI1640完全培养液调整细胞浓度为4×105个/mL。
小鼠脾细胞悬液的制备(效应细胞):方法同(5)。
NK细胞活性检测:取靶细胞和效应细胞各50μL(效靶比50:1),加入U型96孔培养板中;靶细胞自然释放孔加靶细胞和培养液各50μL,靶细胞最大释放孔加靶细胞和2.5%聚乙二醇辛基苯基醚各50μL。设对照组,复方保健胶囊低、中、高剂量组;各组又分单独处理组和联合ConA组;每孔加入供试品100μL,于37℃、5%CO2培养箱中培养24h,然后将96孔培养板以1500r/min离心5min,每孔吸取上清120μL置平底96孔培养板中,同时加入乳酸脱氢酶检测工作液60μL,混匀,室温(约25℃)避光孵育30min(用铝箔包裹后置于水平摇床上缓慢摇动)。然后在490nm处测定吸光度。计算NK细胞活性。
NK细胞活性/%=(反应孔OD-自然释放孔OD)/(最大释放孔OD-自然释放孔OD)
数据分析各组数据用表示,采用SPSS 17.0统计软件进行分析,以P<0.05为差异有统计学意义。
结果与分析
表1:本发明复方保健胶囊对小鼠免疫器官重量及脏器系数测定结果
注:与阴性对照组比较,**表示差异极显著,P<0.01。
与阴性对照组比较,复方保健胶囊各剂量组小鼠脾脏/体重比值均无统计学意义(P>0.05);低、中剂量组的胸腺/体重比值有显著差异(P<0.01);复方保健胶囊各剂量组小鼠肝脏/体重比值均明显下降,其中中剂量组有显著差异(P<0.01)。
表2:本发明复方保健胶囊二硝基氯苯诱导的小鼠耳肿胀实验结果
与阴性对照组比较,在引发超敏反应后,复方保健胶囊高剂量组小鼠右耳肿胀明显。中、低剂量组肿胀度有下降趋势,尤其中剂量组肿胀度下降趋势明显,但也不具统计学意义(P>0.05)。
表3:本发明复方保健胶囊碳粒廓清实验结果
注:与阴性对照组比较,*表示差异显著,P<0.05。
与阴性对照组相比,复方保健胶囊各剂量组小鼠的吞噬指数均增加,中剂量组差异有统计学意义(P<0.05)。表明复方保健胶囊能提高小鼠碳廓清能力,单核-巨噬细胞功能实验结果阳性。
表4:本发明复方保健胶囊脾淋巴细胞增殖能力的测定结果
注:各给药组与正常细胞组比较,**表示差异极显著,P<0.01。ConA处理组与正常细胞组比较,#表示差异显著,P<0.05,##表示差异极显著,P<0.01。
如表4所示,单独给药时,复方保健胶囊具有明显的促淋巴细胞增殖作用,并且这一作用呈浓度依赖性增加。ConA联合处理组对小鼠淋巴细胞增殖有一定的促进作用,高剂量组与对照组相比有显著性差异。联合给药时,复方保健胶囊各剂量组刺激脾淋巴细胞增殖程度均较单独给药时没有显著性差异,提示其与ConA没有协同促进作用。
表5:本发明复方保健胶囊NK细胞活性测定结果
注:联合ConA处理组与正常细胞组比较,*表示差异显著,P<0.05。
在对NK细胞活性测定试验中,与阴性对照组相比,复方保健胶囊单独处理组及联合给药高、中剂量组对小鼠NK细胞活性无显著影响,复方保健胶囊单独处理及联合给药低剂量组有增加NK细胞活性趋势,且联合给药低剂量组与对照组存在显著性差异。
综上,经过试验验证,本发明制备的复方保健胶囊能够提高小鼠胸腺指数,降低小鼠耳肿胀度,促进淋巴细胞增殖,增加小鼠吞噬指数和NK细胞活性。表明复方保健胶囊具有增强免疫力功能,可用于开发增强免疫力作用的保健食品。
上述的对实施例的描述是为便于该技术领域的普通技术人员能理解和使用发明。熟悉本领域技术的人员显然可以容易地对这些实施例做出各种修改,并把在此说明的一般原理应用到其他实施例中而不必经过创造性的劳动。因此,本发明不限于上述实施例,本领域技术人员根据本发明的揭示,不脱离本发明范畴所做出的改进和修改都应该在本发明的保护范围之内。
Claims (10)
1.一种提高免疫力功能的保健组合物,其特征在于,所述组合物包括以下质量分:烟酰胺单核苷酸2.5~3.5份,灵芝孢子粉1.0~1.5份,铁皮石斛粉1.0~1.5份。
3.根据权利要求2所述的一种具有提高免疫力功能的保健组合物胶囊,其特征在于,所述保健组合物囊包括以下重量百分比计的制备原料:
NMN 25%~35%
灵芝孢子粉 10%~15%
铁皮石斛粉 10%~15%
黏合剂、稀释剂 30%~40%
防潮剂、助流剂 8%~12%
润滑剂 2%~3%。
4.根据权利要求2所述的一种具有提高免疫力功能的保健组合物胶囊,其特征在于,所述黏合剂、稀释剂均为甘露醇、微晶纤维素、乳糖、淀粉中的一种或多种的组合。
5.根据权利要求4所述的一种具有提高免疫力功能的保健组合物胶囊,其特征在于,所述黏合剂、稀释剂均为微晶纤维素。
6.根据权利要求2所述的一种具有提高免疫力功能的保健组合物胶囊,其特征在于,所述防潮剂、助流剂均为微粉硅胶、氢化植物油中的一种或者两者的混合。
7.根据权利要求6所述的一种具有提高免疫力功能的保健组合物胶囊,其特征在于,所述防潮剂、助流剂均为微粉硅胶。
8.根据权利要求2所述的一种具有提高免疫力功能的保健组合物胶囊,其特征在于,所述润滑剂为硬脂酸镁、硬脂富马酸钠、单硬脂酸甘油酯中的一种或多种的组合。
9.一种权利要求2至8中任意一项所述保健组合物胶囊的制备方法,其特征在于,包括以下步骤:
(1)将NMN原料药过筛备用,铁皮石斛研磨成细粉,按比例分别称取NMN、灵芝孢子粉、铁皮石斛粉、黏合剂、稀释剂,防潮剂、助流剂,润滑剂;
(2)将NMN原料药、灵芝孢子粉和铁皮石斛粉过筛后与步骤(1)中称取的防潮剂、助流剂按等量递加法混匀,制得混合物料A;
(3)将混合物料A与步骤(1)中称取的黏合剂、稀释剂按等量递加法混匀,过筛,制得混合物料B;
(4)将混合物料B与步骤(1)中称取的润滑剂混合均匀,混合3-5min,制得混合物料C,在胶囊板上进行胶囊填充,制得NMN胶囊成品。
10.根据权利要求9所述的保健组合物胶囊的制备方法,其特征在于,步骤(1)中所述NMN原料药经过筛后的粒径D≤0.250mm,灵芝孢子粉和铁皮石斛粉研磨成细粉过筛后的粒径D≤0.30mm。
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