CN113785975A - 精胺、亚精胺脂质体在抗氧化、抗衰老中的应用 - Google Patents
精胺、亚精胺脂质体在抗氧化、抗衰老中的应用 Download PDFInfo
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- CN113785975A CN113785975A CN202111054501.4A CN202111054501A CN113785975A CN 113785975 A CN113785975 A CN 113785975A CN 202111054501 A CN202111054501 A CN 202111054501A CN 113785975 A CN113785975 A CN 113785975A
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- Prior art keywords
- spermine
- spermidine
- liposome
- solution
- lipid
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Abstract
本发明涉及一种精胺和/或亚精胺脂质体在抗氧化、抗衰老中的应用。它是以精胺和/或亚精胺为有效活性成分。精胺、亚精胺包裹于脂质体中,其原料的质量百分比组成为:精胺或亚精胺0.01~10%;脂质材料0.01~30%;余量为水溶液。本发明提供的精胺或亚精胺脂质体形态规则,平均粒径较小,分布均匀,具有良好的清除DPPH自由基、抗氧化、抗衰老、清除脂褐素和透皮吸收的能力。制得的脂质体能提高精胺、亚精胺的稳定性和透皮吸收性能,制备食用、药用或美妆用产品,尤其是透皮药物制剂和制备美妆产品,所制得的产品具有抗氧化和抗衰老功效且效果好等优点。
Description
技术领域
本发明涉及一种精胺、亚精胺脂质体在抗氧化、抗衰老中的应用,具体涉及一种精胺、亚精胺脂质体的制备工艺,并将其作为活性成分在制备食用、药用或美妆用组合物中的抗氧化、抗衰老的用途。
背景技术
精胺[Spermin, SPM, N,N’-双(3-氨基丙基)-1,4-丁二胺]与亚精胺[Spermidine, SPMD, N-(3-氨基丙基)-1,4-丁二胺]都属于脂肪族聚胺类化合物。因其最初是在人的精液中被发现而得名,存在于细菌和大多数动物细胞中,在调控细胞生长、分裂、分化和动物组织增殖等方面发挥关键作用,是重要的细胞生长和增殖因子。药理学研究表明,精胺、亚精胺还具有抗氧化、延缓衰老、抗炎和免疫调节等功效。因此,将精胺、亚精胺开发成功能性食用、药用或美妆产品,用于抗氧化、抗衰老具有重要的现实意义。精胺、亚精胺等多胺类成分广泛存在于日常饮食中,且多胺的保健功效作用正在逐渐被科学所证实,随着研究的不断深入,其应用将日益广泛,故将多胺类成分应用于食用、药用和美妆产品领域发挥其抗氧化、抗衰老的功效是安全且可行的。制备工艺便捷,存储条件简单且使用方便的上述产品,符合市场的需求。
精胺和亚精胺等多胺类成分具有优良的性质和功效,目前的研究多关注在精胺、亚精胺单体及其功能组合物的美白、对抗皮肤衰老等功效。但精胺和亚精胺在应用中存在局限性,如透皮吸收能力差,不利于人体吸收和在美妆产品中应用等。寻找理想优质的载体递送精胺、亚精胺,有效提高精胺、亚精胺的透皮吸收能力具有重要的现实意义。
中国专利CN109952096A公开了聚胺在用于诱导或促使皮肤变黑并调节黑素原生成的组合物和方法中的用途,所述方法包括施用多阳离子脂肪族胺,其中所述多阳离子脂肪族胺优选地为腐胺、亚精胺和精胺。
脂质体是由磷脂、胆固醇等类脂质物质通过自组装形成具有双分子薄层结构的超微球状体,其粒径一般在几十纳米到几微米之间。脂质体由类脂质双分子层组成的膜将内外环境隔开,膜表面由亲水基团组成,膜内部由疏水链聚集而成,脂质体既能负载亲脂性物质,也能负载亲水性物质,利用脂质体可以和细胞膜融合的特点,将某些药物或化学成分递送进入细胞内部。脂质体具有被动靶向的特点,能够缓慢地释放药物,延长药物作用时间,提高药物或化学成分的稳定性,促进药物透皮吸收,对局部疾病部位的靶向,降低药物或化学成分的毒性和刺激性等特点。因此,脂质体作为药物载体已广泛应用于药物制剂及美妆产品工艺配方中。制备精胺、亚精胺脂质体可提高精胺、亚精胺的稳定性、穿透皮肤的能力和缓释的性能。因此,制备精胺、亚精胺脂质体及其在食用、药用和美妆产品中的应用将成为研究热点。
发明内容
本发明的目的在于提供一种精胺、亚精胺脂质体和制备方法及精胺、亚精胺脂质体在抗氧化、抗衰老中的应用。本发明提供的精胺、亚精胺脂质体形态规则,平均粒径较小,分布均匀,具有良好的清除DPPH自由基、抗氧化、抗衰老、清除脂褐素和透皮吸收的能力,提高了精胺、亚精胺的稳定性和透皮性能,弥补了精胺、亚精胺在制备食用、药用或美妆用产品中的不足,使其在上述产品中的应用更为方便合理和方便。只需将其与配方中的其他成分混合均匀即可。
本发明提供的一种精胺、亚精胺脂质体,它是以精胺、亚精胺为有效活性成分,精胺、亚精胺包裹于脂质体中,其原料的质量百分比组成为:
精胺或亚精胺 0.01~10%
脂质材料 0.01~30%
余量为水溶液
所述的脂质材料为大豆卵磷脂、蛋黄卵磷脂或胆固醇中的一种或多种。
上述精胺或亚精胺的质量百分比优选为0.01~5%。
脂质材料的质量百分比优选为0.01~10%。
所述的水溶液为磷酸缓冲液、蒸馏水、去离子水、纯净水中的一种;其中,磷酸缓冲液的pH值在7。
本发明提供的一种精胺、亚精胺脂质体的制备方法包括下述步骤:
1)按计量将卵磷脂和胆固醇溶于有机溶剂,卵磷脂与胆固醇摩尔比为1:1~10,得到脂质溶液,40℃减压下旋转蒸发除去有机溶剂,使之在瓶壁上形成均匀的磷脂薄膜,在真空干燥器中放置12-16小时。
2)加入0.3 mol/L柠檬酸缓冲溶液(pH = 4.0)作为水合介质,在浴式超声仪中超声10 min,探头超声9 min(每隔3 min停止一次),室温下水合磷脂膜,制备空白脂质体,将所得脂质体通过0.22 μm微孔滤膜,进行整粒。
3)将上述空白脂质体溶液加入凝胶柱(2 cm × 60 cm),用磷酸盐缓冲溶液(pH =7.4)进行洗脱,收集洗脱后的脂质体溶液,按照计量的药脂比(药物质量/磷脂质量)加入精胺或亚精胺,空气振荡器中振荡1 h,4 ℃条件下孵育一定时间,即得精胺或亚精胺脂质体。
所述的精胺或亚精胺脂质体是将精胺或亚精胺包裹于脂质体的内部,形成一种亲水性的乳白色混悬液。
步骤1)中所述的有机溶剂为乙醇、二氯甲烷、三氯甲烷或乙醚,优选为三氯甲烷。所述卵磷脂与胆固醇摩尔比优选为1: 1~3。
步骤1)中,制成的脂质溶液的浓度为0.1~20 g/100mL。
步骤3)中所述的药脂比优选为1:10~60,更优选为1:20~30。
步骤3)中所述的药脂比中,精胺或亚精胺的质量含量为1%~20%。
本发明提供了上述精胺或亚精胺脂质体的用途。将其作为活性成分用于制备食用、药用或美妆用产品,所述产品包括胶囊、片剂、口服液、颗粒剂、保健饮品、保健酒、化妆水、面霜、精华、乳液或霜剂。
在上述方案的基础上,所述的精胺和/或亚精胺脂质体在食用、药用或美妆用产品中的质量百分含量为0.5%~10%。
本发明公开的精胺、亚精胺脂质体具有良好的清除DPPH自由基、抗氧化、抗衰老、清除脂褐素和透皮吸收的能力,提高了精胺、亚精胺的稳定性和透皮性能,弥补了精胺、亚精胺在制备食用、药用或美妆用产品中的不足,使其在上述产品中的配制更为方便合理和方便。只需将其与配方中的其他成分混合均匀即可。
本发明所述的精胺、亚精胺脂质体具有的主要优点为:
1)提高了精胺、亚精胺的稳定性。如果仅是简单的将药物与其他基质混合在一起,由于药物间的相互作用、光线、氧气、酸、碱等的影响,在制备和储藏过程中一部分便失去了活性。将精胺、亚精胺包裹在脂质体中,可避免药物间的相互作用及各种不稳定因素对药物的破坏,从而提高了其稳定性。
2)增加了精胺、亚精胺的皮肤通透性。脂质结构类似生物膜,其与皮肤角质层的磷脂相互作用,改变角质层对药物的屏障功能,通过吸附、融合、水合的方式将药物释放到皮肤中,从而提高了药物的皮肤穿透性。
3)改善了精胺、亚精胺的释放性能。脂质体进入机体后(包括皮肤等),可以在血液、表皮、真皮等形成药物储库,使药物缓慢释放,在细胞内外直接、持久地发挥作用。
附图说明
图1 实施例1中本发明制备的精胺、亚精胺脂质体透射电镜图片。
图2 实施例2中精胺、亚精胺脂质体对DPPH自由基的清除能力。
图3 实施例3中精胺、亚精胺脂质体的总抗氧化能力。
图4 实施例4中精胺、亚精胺脂质体透皮吸收的能力。
图5 实施例5中精胺、亚精胺脂质体对线虫头部摆动频率的影响。
图6 实施例6中精胺、亚精胺脂质体对线虫吞咽频率的影响。
图7 实施例7中精胺、亚精胺脂质体对线虫体内脂褐素含量的影响。
具体实施方式
下面结合实施例及附图对本发明作进一步详细的描述,但本发明的实施方式不限于此。
主要试剂及材料来源:大豆卵磷脂(大连美仑生物技术有限公司);胆固醇(Sigma-Aldrich公司,美国);精胺、亚精胺(上海阿拉丁生化科技股份有限公司,CAS分别为71-44-3和124-20-9);DPPH自由基清除能力试剂盒、总抗氧化能力(T-AOC)测试盒(ABTS法)均购自南京建成生物工程研究所;96孔板购自Corning;酶标仪(伯乐生命医学产品(上海)有限公司);Franz透皮扩散试验仪(型号RYJ-12B,北京铭成基业科技有限公司);BSA124S精密电子天平(德国赛多利斯);超纯水仪(默克密理博Milli-Q,德国);粒径-Zeta电位分析仪(Zetasizer Nano-ZS90,Malvern,英国);透射电子显微镜(Hitachi HT7700,日本);透析袋(截留分子量为500 Da,北京博奥拓达科技有限公司);台式恒温振荡器(天津市欧诺仪器仪表有限公司);C18色谱柱(4.6×150 mm,5 µm,GL Sciences公司,日本);Agilent 1260高效液相色谱仪(安捷伦,美国);酶标仪(Thermo Scientific Multiskan GO,美国);野生型秀丽隐杆线虫(品系Bristol N2)与尿嘧啶缺失型大肠杆菌均购于美国明尼苏达大学的线虫中心(Caenorhabditis Genetics Center, CGC)。
实施例1:精胺、亚精胺脂质体制剂的制备,所述脂质体制剂通过如下方法制备:
1)称取大豆卵磷脂25 mg和胆固醇6.88 mg溶于20mL氯仿,得到脂质溶液,40℃减压下旋转蒸发除去氯仿,使之在瓶壁上形成均匀的磷脂薄膜,放置在真空干燥器中12小时。
2)加入0.3 mol/L柠檬酸缓冲溶液(pH = 4.0)作为水合介质,在浴式超声仪中超声10 min(200 W),探头超声9 min(100 W)(每隔3 min停止一次),室温下水合磷脂膜,制备空白脂质体,将所得脂质体通过0.22 μm微孔滤膜,进行整粒。
3)将上述空白脂质体溶液加入Sephadex G-50凝胶柱(2 cm × 60 cm)(北京索莱宝科技有限公司),用磷酸盐缓冲溶液(pH = 7.4)进行洗脱,收集洗脱后的脂质体溶液,按照一定的药脂比(药物质量/磷脂质量)加入精胺或亚精胺0.625 mg,在空气振荡器中振荡1h,4℃条件下孵育一定时间,即得精胺脂质体(含药量为9.53%)、亚精胺脂质体(含药量为18.15%)。
取30 μL本发明制备的脂质体,分散于970 μL超纯水中,充分混匀后,通过粒径-Zeta电位分析仪检测纳米体系的粒径、PDI以及Zeta电位,重复测量3次,每次1 min。
用移液器吸取20 μL本发明制备的脂质体滴于铜网上,5 min后用滤纸吸去多余液体,待铜网上的水分挥去后即可通过透射电子显微镜观察纳米体系的形貌特征。
通过HPLC建立精胺、亚精胺浓度的检测方法。待测样品经丹磺酰氯衍生化反应,得到样品溶液进行检测:进样量为5 μL,流动相为水/乙腈=20/80(v/v),流速为1 mL/min,柱温为30℃,检测器波长为254 nm的色谱条件下检测。建立精胺与亚精胺浓度相对于其峰面积的标准曲线,并计算精胺、亚精胺纳米脂质体的载药量与包封率,计算公式如下:载药量=脂质体中精胺或亚精胺的质量/纳米脂质体的质量×100%;包封率=脂质体中精胺或亚精胺的质量/精胺或亚精胺的投量×100%。
实验结果如图1所示,通过粒径-Zeta电位分析仪测定精胺、亚精胺脂质体的粒径分别为347.4±38.7 nm和334.8±37.9 nm,PDI分别为0.250±0.039和0.229±0.050,Zeta电位分别为-49.7±4.2 mV和-42.5±3.6.9 mV。通过TEM进一步观察精胺、亚精胺脂质体的形貌特征,如图1所示,制备的脂质体呈规则球形,具有核壳结构,分散性良好。通过HPLC的峰面积建立精胺、亚精胺浓度的标准曲线,可进一步通过计算得精胺、亚精胺脂质体中精胺与亚精胺的载药量。
通过调整大豆卵磷脂、胆固醇、精胺、亚精胺之间的比例,可制备载药量在5%-20%的精胺、亚精胺脂质体,实现精胺、亚精胺的有效负载。
使用实施例1制备的精胺、亚精胺脂质体进行下述实验。
实施例2:精胺、亚精胺脂质体DPPH自由基清除能力测定
1,1-二苯基-2-苦基肼基自由基[1,1-Diphenyl-2-picrylhydrazyl radical,DPPH]广泛用于定量测定生物试样的抗氧化能力。根据DPPH自由基有单电子,在517 nm处有一强吸收,其醇溶液呈紫色的特性,当有自由基清除剂存在时,由于与其单电子配对而使其吸收逐渐消失,呈现的颜色越浅,即吸光度A值越低,进而对样本中DPPH清除能力进行定量分析。参照DPPH自由基清除能力试剂盒说明书,设对照管、测定管、空白管。其中,对照管:依次加入400 μL不同浓度的受试样品(实施例1制备的精胺、亚精胺脂质体及游离精胺、亚精胺),600 μL 80%甲醇。测定管:依次加入400 μL不同浓度的受试样品,600 μL工作液。空白管:依次加入400 μL 80%甲醇,600 μL工作液。各管混匀后,室温25 ℃避光静置30 min,4000 rad/min离心5 min,取上清液800 μL至比色皿中,无水甲醇调零,517 nm波长检测各管吸光度A值。按下式分别计算各样品对DPPH自由基的清除率。DPPH自由基清除率(%)=[1-(A测定-A对照)/A空白]×100 %
实验结果如图2所示,与空白对照组比较,精胺、亚精胺体外均显示出良好的清除DPPH自由基的能力(P<0.05),精胺、亚精胺体外清除DPPH自由基的能力差异无统计学意义(P>0.05)。与空白脂质体组比较,精胺脂质体、亚精胺脂质体体外均显示出良好的清除DPPH自由基的能力(P<0.05),精胺脂质体、亚精胺脂质体体外清除DPPH自由基的能力差异无统计学意义(P>0.05)。与精胺组比较,精胺脂质体体外清除DPPH自由基的能力差异无统计学意义(P>0.05)。与亚精胺组比较,亚精胺脂质体体外清除DPPH自由基的能力差异无统计学意义(P>0.05)。
实施例3:精胺、亚精胺脂质体总抗氧化能力测定(ABTS法)
ABTS在适当的氧化剂作用下氧化成绿色的ABST·+,在抗氧化物存在时,ABST·+的产生会被抑制,在405 nm或734 nm测定ABST·+的吸光度即可测定并计算样品的总抗氧化能力。Trolox是一种VE的类似物,具有和VE相近的抗氧化能力,用作其它抗氧化物总抗氧化能力的参考。例如,Trolox的总抗氧化能力为1,相同浓度情况下,其它物质的抗氧化能力用其抗氧化能力和Trolox相比的倍数来表示。参照总抗氧化能力(T-AOC)试剂盒说明书,采用96孔板检测,设空白孔、标准孔及测定孔。空白孔:依次加入10 μL蒸馏水,20 μL应用液,170 μLABTS工作液;标准孔:依次加入10 μLTrolox标准液,20 μL应用液,170 μL ABTS工作液;测定孔:依次加入10 μL待测样本(实施例1制备的精胺、亚精胺脂质体及游离精胺、亚精胺),20 μL应用液,170 μLABTS工作液。混匀后,室温反应6 min,波长405 nm,酶标仪读取各孔OD值。以标准品OD值为横坐标,各OD值对应的标准品浓度为纵坐标制成标准曲线,EXCEL制得曲线公式,把样本测定管测得的OD代入计算公式,求得结果。
实验结果如图3所示,与空白对照组比较,精胺与亚精胺体外均显示出良好的体外抗氧化能力(P<0.05),精胺与亚精胺体外抗氧化的能力差异无统计学意义(P>0.05)。与空白脂质体组比较,精胺脂质体与亚精胺脂质体均显示出良好的体外抗氧化能力(P<0.05),精胺脂质体与亚精胺脂质体体外抗氧化能力差异无统计学意义(P>0.05)。与精胺组比较,精胺脂质体体外抗氧化能力差异无统计学意义(P>0.05)。与亚精胺组比较,亚精胺脂质体体外抗氧化能力差异无统计学意义(P>0.05)。
实施例4:精胺和/或亚精胺脂质体透皮吸收能力检测
采用Franz透皮扩散试验仪对实施例1制备的精胺、亚精胺脂质体及游离精胺、亚精胺透皮吸收能力进行检测。取SD大鼠皮肤:采用10%水合氯醛麻醉大鼠,剃毛器小心剃去大鼠腹部的毛,剃毛刀除去余毛。脱颈椎处死后,剥离其腹部皮肤,采用60℃的热水浸泡40s,除去皮下脂肪组织,采用手术刀片小心刮去皮下筋膜,得到大鼠皮肤,经无菌生理盐水清洗后使用。将大鼠皮肤固定于Franz扩散池之间(扩散池有效扩散面积为2.2 cm2,接受池体积为8 mL);精密量取1mL溶液(包括空白脂质体、精胺脂质体、亚精胺脂质体、精胺溶液、亚精胺溶液,其中精胺或亚精胺等量浓度为30mg/mL),使其完全贴于大鼠皮肤表面进行透皮吸收实验;接收液为去离子水,温度设为32±0.2 ℃,接收池搅拌子转速为500 rpm;定时(1、2、4、6、8、10、12、24 h)取出全部接收液,备用,同时补充新鲜接收液。采用丹磺酰氯对接受液进行衍生化处理。建立精胺和亚精胺浓度检测标准标准曲线。将上述经衍生化反应得到的标准溶液在进样量为5 μL,流动相为水:乙腈=20:80,流速为1 mL/min,柱温为30℃,检测器波长为254 nm的色谱条件下检测。
实验结果如图4所示,以纵坐标为待测成分不同时间点的累计透过百分比(%),横坐标为取样时间t,绘制累计透过百分比(%)-时间曲线。实验数据表明,精胺和亚精胺以及精胺脂质体和亚精胺脂质体单位面积累积透皮渗透量均随时间的延长而逐渐增加。精胺和亚精胺脂质体24 h单位面积累计透皮渗透量明显高于精胺和亚精胺。
实施例5:精胺、亚精胺脂质体对线虫头部摆动频率的影响
随着年龄的增长,线虫在固体和液体介质上的运动能力下降,这与肌肉退化的程度有关,因此线虫运动能力下降常作为评价衰老的指标之一。头部摆动频率可显示出线虫运动能力的强弱。采用线虫模型评价实施例1制备的精胺、亚精胺脂质体及游离精胺、亚精胺对线虫头部摆动频率的影响。线虫头部摆动频率测定实验按以下方法分组:空白对照组只加入适量的灭菌后的纯水;实验组分别为5 mM 精胺干预组、5 mM 亚精胺干预组、5 mM精胺脂质体干预组、5 mM 亚精胺脂质体干预组、空白脂质体干预组。每隔1天将它们转移到已标记的相对应的各浓度干预组的培养基中。实验第9天,挑取不同干预组的线虫放于M9缓冲液中,在观察记录头部摆动频率前,使线虫在液体中适应1 min,然后再记录1 min内线虫的头部摆动次数。线虫头部两次经过身体中线记为1次头部摆动。每次实验随机挑选10条线虫,重复3次。
实验结果如图5所示,在显微镜下,通过计算线虫在液体中每分钟头部摆动的次数来观察精胺和亚精胺脂质体对N2线虫运动行为的影响。实验数据表明,空白脂质体与空白对照组比较,两组线虫摆动的次数的差异无统计学意义(P>0.05)。与空白对照组比较,精胺、亚精胺均可明显增加线虫摆动的次数(P<0.05)。与空白脂质体组比较,精胺脂质体、亚精胺脂质体均可明显增加线虫摆动的次数(P<0.05)。分别与精胺、亚精胺组比较,精胺脂质体、亚精胺脂质体增加线虫摆动的次数明显优于精胺、亚精胺(P<0.05)。由此可见,精胺与亚精胺纳米脂质体可明显改善线虫的运动能力,在延缓衰老的过程中可能发挥一定的作用,且作用强度强于精胺与亚精胺。
实施例6:精胺、亚精胺脂质体对线虫吞咽频率的影响
采用线虫模型评价实施例1制备的精胺、亚精胺脂质体及游离精胺、亚精胺对线虫吞咽频率的影响。分组同实施例5,同步化后生长至L4期的线虫分别放置到对照组及含各种干预物的NGM平板中。在开始干预第6天,每组随机挑取10条线虫,放置在20 μL的M9 缓冲液中,记录1 min内的线虫吞咽次数。每个浓度重复三次。
实验结果如图6所示,空白脂质体与空白对照组比较,两组线虫的咽泵运动速率差异无统计学意义(P>0.05)。与空白对照组比较,精胺、亚精胺均可明显提高线虫的咽泵运动速率(P<0.05)。与空白脂质体组比较,精胺脂质体、亚精胺脂质体均可明显提高线虫的咽泵运动速率(P<0.05)。由此可见,精胺与亚精胺纳米脂质体可明显提高线虫的咽泵运动速率,改善线虫的肌肉运动功能,延缓衰老。
实施例7:精胺和/或亚精胺脂质体对线虫体内脂褐素含量的影响
随着线虫衰老进程的发展,脂褐素会在其肠道内逐渐积累,且自发荧光,是线虫衰老的有效生物标志。采用线虫模型评价实施例1制备的精胺、亚精胺脂质体及游离精胺、亚精胺对线虫体内脂褐素含量的影响。空白对照组只加入适量的灭菌后的纯水;实验组分别为5 mM 精胺干预组、5 mM 亚精胺干预组、5 mM 精胺脂质体干预组、5 mM 亚精胺脂质体干预组、空白脂质体干预组。每组挑取30条同期化线虫置于对应的培养基内,干预5天后进行脂褐素荧光检测。将线虫分别挑至滴加左旋咪唑溶液(20 μL, 0.1mM)的琼脂糖垫上进行麻醉,使用倒置荧光显微镜,固定曝光时间和强度,激发波长设置为340-380 nm,发射波长为430 nm,观察线虫体内的脂褐素荧光强度,并拍摄荧光图片,用Image J软件测量荧光强度值。
实验结果如图7所示,空白脂质体与空白对照组比较,两组线虫的咽泵运动速率差异无统计学意义(P>0.05)。与空白对照组比较,精胺、亚精胺均可明显提高线虫的咽泵运动速率(P<0.05)。与空白脂质体组比较,精胺脂质体、亚精胺脂质体均可明显提高线虫的咽泵运动速率(P<0.05)。由此可见,精胺与亚精胺纳米脂质体可明显提高线虫的咽泵运动速率,改善线虫的肌肉运动功能,延缓衰老。
以上所述,仅为本发明较佳的具体实施方式,但本发明的保护范围并不局限于此,任何熟悉本技术领域的技术人员在本发明披露的技术范围内,根据本发明的技术方案及其发明构思加以等同替换或改变,都应涵盖在本发明的保护范围之内。
Claims (9)
1.一种精胺或亚精胺脂质体,其特征在于它是以精胺或亚精胺为有效活性成分,精胺或亚精胺包裹于脂质体中,其原料的质量百分比组成为:
精胺或亚精胺 0.01~10%
脂质材料 0.01~30%
余量为水溶液
所述的脂质材料为大豆卵磷脂、蛋黄卵磷脂或胆固醇中的一种或多种;
所述的水溶液为磷酸缓冲液、蒸馏水、去离子水、纯净水中的一种。
2.根据权利要求1所述的精胺、亚精胺脂质体,其特征在于所述的精胺或亚精胺的质量百分比为0.01~5%;所述的脂质材料的质量百分比为0.01~10%;所述的磷酸缓冲液的pH值为7。
3.权利要求1所述的精胺或亚精胺脂质体的制备方法,其特征在于包括下述步骤:
1)按计量将卵磷脂和胆固醇溶于有机溶剂,卵磷脂与胆固醇摩尔比为1:1~10,得到脂质溶液,40℃减压下旋转蒸发除去有机溶剂,使之在瓶壁上形成均匀的磷脂薄膜,在真空干燥器中放置12-16小时;
2)加入0.3 mol/L柠檬酸缓冲溶液(pH = 4.0)作为水合介质,在浴式超声仪中超声10min,探头超声9 min,每隔3 min停止一次,室温下水合磷脂膜,制备空白脂质体,将所得脂质体通过0.22 μm微孔滤膜,进行整粒;
3)将上述空白脂质体溶液加入2 cm × 60 cm的凝胶柱,用pH = 7.4磷酸盐缓冲溶液进行洗脱,收集洗脱后的脂质体溶液,按照计量的药脂比加入精胺或亚精胺,空气振荡器中振荡1 h,4 ℃条件下孵育,即得精胺或亚精胺脂质体。
4.根据权利要求3所述的制备方法,其特征在于步骤1)中所述的有机溶剂为乙醇、二氯甲烷、三氯甲烷或乙醚,优选为三氯甲烷;所述卵磷脂与胆固醇摩尔比优选为1: 1~3。
5.根据权利要求3所述的制备方法,其特征在于:步骤1)中,制成的脂质溶液的浓度为0.1~20 g/100mL。
6.根据权利要求3所述的制备方法,其特征在于:步骤3)中所述的药脂比优选为1:10~60,更优选为1:20~30。
7.根据权利要求3所述的制备方法,其特征在于:步骤3)中所述的药脂比中,精胺或亚精胺的质量含量在1%~20%。
8.权利要求1所述的精胺或亚精胺脂质体的用途,将其作为活性成分用于制备食用、药用或美妆用产品,所述产品包括胶囊、片剂、口服液、颗粒剂、保健饮品、保健酒、化妆水、面霜、精华、乳液或霜剂。
9.根据权利要求7所述的的用途,其特征在于所述的精胺和/或亚精胺脂质体在食用、药用或美妆用产品中的质量百分含量为0.5%~10%。
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Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20020001612A1 (en) * | 1996-11-12 | 2002-01-03 | The Regents Of The University Of California | Methods of forming protein-linked lipidic microparticles, and compositions thereof |
CN1829496A (zh) * | 2003-07-31 | 2006-09-06 | 朱利亚尼股份公司 | 精胺和/或亚精胺在食用、药用或美容用组合物中对抗皮肤老化的用途 |
US7491409B1 (en) * | 1999-03-02 | 2009-02-17 | Transave, Inc. | Encapsulation of bioactive complexes in liposomes |
US20100215582A1 (en) * | 2007-05-14 | 2010-08-26 | Konica Minolta Holdings, Inc. | Liposome and method for producing liposome |
CN102085157A (zh) * | 2011-01-27 | 2011-06-08 | 中国水产科学研究院黄海水产研究所 | 扇贝抗氧化肽纳米脂质体及其制备方法 |
CN104876831A (zh) * | 2015-04-03 | 2015-09-02 | 苏州圣诺生物医药技术有限公司 | 脂质修饰精胺衍生物及利用该衍生物制备的脂质体 |
CN104997652A (zh) * | 2015-07-09 | 2015-10-28 | 西安艾尔菲生物科技有限公司 | 一种抗皱保湿脂质体及其制备方法和应用 |
CN109453117A (zh) * | 2018-10-26 | 2019-03-12 | 南京中医药大学 | 强心苷类活性化合物脂质体及其制备方法 |
CN109952096A (zh) * | 2016-09-08 | 2019-06-28 | 新加坡科技研究局 | 聚胺在用于诱导或促使皮肤变黑并调节黑素原生成的组合物和方法中的用途 |
US20220396556A1 (en) * | 2019-10-18 | 2022-12-15 | The Trustees Of The University Of Pennsylvania | Lipid and Lipid Nanoparticle Formulation for Drug Delivery |
-
2021
- 2021-09-09 CN CN202111054501.4A patent/CN113785975B/zh active Active
Patent Citations (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20020001612A1 (en) * | 1996-11-12 | 2002-01-03 | The Regents Of The University Of California | Methods of forming protein-linked lipidic microparticles, and compositions thereof |
US7491409B1 (en) * | 1999-03-02 | 2009-02-17 | Transave, Inc. | Encapsulation of bioactive complexes in liposomes |
CN1829496A (zh) * | 2003-07-31 | 2006-09-06 | 朱利亚尼股份公司 | 精胺和/或亚精胺在食用、药用或美容用组合物中对抗皮肤老化的用途 |
US20060252838A1 (en) * | 2003-07-31 | 2006-11-09 | Fabio Rinaldi | Use of spermine and/or spermidine against skin ageting in dietary, pharmaceuetical or cosmetic compositions |
US20100215582A1 (en) * | 2007-05-14 | 2010-08-26 | Konica Minolta Holdings, Inc. | Liposome and method for producing liposome |
CN102085157A (zh) * | 2011-01-27 | 2011-06-08 | 中国水产科学研究院黄海水产研究所 | 扇贝抗氧化肽纳米脂质体及其制备方法 |
CN104876831A (zh) * | 2015-04-03 | 2015-09-02 | 苏州圣诺生物医药技术有限公司 | 脂质修饰精胺衍生物及利用该衍生物制备的脂质体 |
CN104997652A (zh) * | 2015-07-09 | 2015-10-28 | 西安艾尔菲生物科技有限公司 | 一种抗皱保湿脂质体及其制备方法和应用 |
CN109952096A (zh) * | 2016-09-08 | 2019-06-28 | 新加坡科技研究局 | 聚胺在用于诱导或促使皮肤变黑并调节黑素原生成的组合物和方法中的用途 |
CN109453117A (zh) * | 2018-10-26 | 2019-03-12 | 南京中医药大学 | 强心苷类活性化合物脂质体及其制备方法 |
US20220396556A1 (en) * | 2019-10-18 | 2022-12-15 | The Trustees Of The University Of Pennsylvania | Lipid and Lipid Nanoparticle Formulation for Drug Delivery |
Non-Patent Citations (1)
Title |
---|
赵丽等, 厦门大学出版社, pages: 2 * |
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