纳米辅酶Q_（10）抗皮肤光老化及其抗氧化作用研究
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摘要
辅酶Q10是线粒体的能量来源,是体内天然的抗氧化剂,具有重要的生理作用和临床应用价值。但是,辅酶Q10是脂溶性的物质,生物利用度低,限制了其应用。
纳米悬浮液制剂是近年来提出的的新剂型,是纳米载药系统的一种,能够提高药物溶解性、稳定性和生物利用度,还可以实现药物的靶向给药、控制药物释放、增加药物稳定性、降低不良反应、减少用药剂量等特点。
因此,本论文旨评价辅酶Q10纳米混悬液载药系统极其机理,进行了纳米制剂的表征,大鼠体内药代动力学实验,口服辅酶Q10纳米混悬液体内抗氧化实验；还进行了体外鼠皮渗透试验,抗皮肤光老化能力实验。本论文研究成果如下：
1、研究了纳米辅酶Q10质量评价和稳定性通过AFM电镜和激光粒度仪显示辅酶Q10为球形颗粒,粒径在200±50nm,zate电位为-29.8 mV,分散性和稳定性良好的体系。X射线衍射和DSC均显示辅酶Q10以无定形形式存在于纳米悬浮液中。辅酶Q10纳米悬浮液冻干粉中辅酶Q10的饱和溶解度和体外溶出速率都远高于辅酶Q10原药(P<0.05)；
2、研究了纳米辅酶Q10药代动力学特性结果表明,Tmax(3.3±0.23 h)显著小于辅酶Q10原药的Tmax值(P<0.05),表明辅酶Q10经纳米后在大鼠体内的吸收加快,达峰时间提前。纳米化后药物的Cmax (3.23±0.09μg/mL)显著高于原药的Cmax值(P<0.05),AUC值也大于原药的AUC,表明辅酶Q10经纳米化后在大鼠体内能达到更高的血浆浓度,机体生物利用度也显著增加,以原药(R)作为参比制剂,纳米混悬液(T)的相对生物利用度F为(101±3)%；
3、研究了纳米辅酶Q10体内抗氧化活性结果表明,较原药组和阳性对照组,纳米组显著升高血浆SOD、CAT和GSH-Px活力和肝脏的SOD活力(P<0.05),并明显降低小鼠血浆和肝脏MDA含量(P<0.05),并呈现剂量依赖性,辅酶Q10纳米混悬液对小鼠肝脏和肾脏组织超微结构无任何病理学影响,是安全可靠的新剂型；
4、研究了纳米辅酶Q10透皮能力和抗UVB皮肤老化通过体外皮肤渗透实验研究,表明辅酶Q10纳米混悬液具有优良的透皮能力,稳态渗透率由为13.54±1.10μg·cm-2·h-1,是市售商品的5.46倍。在UVB皮肤光老化实验中,较原药组和美丽加芬阳性对照组,纳米混悬液组显著升高血浆和皮肤中SOD、CAT和GSH-Px活力(P<0.05),并明显降低小鼠血浆和皮肤中MDA含量(P<0.05),提高了皮肤中羟脯氨酸的含量(P<0.05)。
综上所述,辅酶Q10纳米混悬液是辅酶Q10无定形分散体系,载药量高,粒子形态大多为圆球形,分散性良好,粒径约为200±50 nm,具有良好的稳定性。辅酶Q10纳米混悬液能够提高药物溶解性、稳定性和生物利用度,增强了药效、降低不良反应、减少用药剂量等特点。
Coenzyme Q10 is mitochondrial energy source and natural antioxidants in organism, having an important physiological role and clinical value. However, the coenzyme Q10 is a fat-soluble substances, with poor human absorption, low bioavailability and poor stability constituting a bottleneck in its formulation and application.
In recent years, nanosuspension is a new nano-drug delivery system, can increase drug solubility, stability and bioavailability,but also achieve targeted drug delivery, controlled drug release, increase drug stability, reduce side effects, reduce the dosage and so on.
Therefore, the aims of the study is to assess coenzyme Q10 suspension containing nano-medicine system from aspects of quality control and the pharmacodynamic. First of all, investigated preparation for the characterization of nano suspensions the quality of coenzyme Q10; we did pharmacokinetics experimental in rats to investigate in vivo absorption and metabolism; to investigate effect of oral coenzyme Q10 nano-suspension on antioxidative enzyme system,we detect the SOD, CAT, GSH-Px activity and MDA content; carried out in vitro rat skin permeation tests, to investigate the penetration ability of coenzyme Q10 nano-suspension, and the establishment of UVB light skin aging model to study nanosuspension coenzyme Q10 to see skin photoaging ability. Thus, for the clinical application of coenzyme Q10, health products, food and cosmetics fields widely used, open up good prospects. The results of the research topics are as follows:
1、Stability of quality assessment and quality evaluation of coenzyme Q10 and stability of nano-suspension Transmission electron microscopy shows that coenzyme Q10 as spherical particles, diameter of 200±50nm, good dispersion and stability of the system. X-ray diffraction and DSC show amorphous form of Coenzyme Q10 present in the nano-suspension. Nano Suspensions of coenzyme Q10,freeze-dried powder in saturated solubility and dissolution rate of coenzyme Q are much higher than the original drug, nanoparticle size could be reduced, increasing the particle surface area, to further increase its solubility;
2、Pharmacokinetics Tmax (3.3±0.23 h) was significantly smaller than the original drug coenzyme Q10 Tmax values (P<0.05), showed that coenzyme Q10 by the nano-absorption in rats after the accelerated time to peak early. Nanocrystalline drug Cmax (3.23±0.09μg/ mL) was significantly higher than the original drug Cmax values (P<0.05), AUC=78±2.49μg/(mL·h) value is also larger than the original drug AUC values that coenzyme Q10 in rats after by the ultra fine powder to achieve a higher plasma concentration, bioavailability body also increased significantly 1 times the original drug;
3、Antioxidant activity in mouse Compared to ordinary powder group and positive control group had significantly higher ultrafine plasma SOD, CAT and GSH-Px activity and liver SOD activity (P<0.05),and significantly reduced plasma and liver MDA content in mice (P<0.05), dose-dependent manner, coenzyme Q10 on the ultrastructure of mouse liver and kidney without any pathological effects of the new formulation is safe and reliable;
4、UVB Skin aging and penetration ability The studies about skin permeation in vitro showed that coenzyme Q10 nano-suspension has good penetration capacity, steady-state penetration rate of Js was 17.54±1.10, is a commercial product is marketed product 12 times. UVB skin aging in the experiment, compared with ordinary powder Li Jiafen positive control group and U.S.group had significantly higher ultrafine plasma and skin in the SOD, CAT and GSH-Px activity (P<0.05),and significantly reduced in mice MDA content in plasma and skin (P<0.05), increased content of hydroxyproline in the skin.
In summary, the coenzyme Q10 nanosuspension is coenzyme Q10 amorphous dispersion, has high drug loading, and particle shape are mostly spherical.well dispersed system with particle size of about 200±50 nm, and has good stability. Coenzyme Q10 nanosuspension increases drug solubility, stability and bioavailability, and increased efficacy, reduces side effects and the dosage and so on.
纳米悬浮液制剂是近年来提出的的新剂型,是纳米载药系统的一种,能够提高药物溶解性、稳定性和生物利用度,还可以实现药物的靶向给药、控制药物释放、增加药物稳定性、降低不良反应、减少用药剂量等特点。
因此,本论文旨评价辅酶Q10纳米混悬液载药系统极其机理,进行了纳米制剂的表征,大鼠体内药代动力学实验,口服辅酶Q10纳米混悬液体内抗氧化实验；还进行了体外鼠皮渗透试验,抗皮肤光老化能力实验。本论文研究成果如下：
1、研究了纳米辅酶Q10质量评价和稳定性通过AFM电镜和激光粒度仪显示辅酶Q10为球形颗粒,粒径在200±50nm,zate电位为-29.8 mV,分散性和稳定性良好的体系。X射线衍射和DSC均显示辅酶Q10以无定形形式存在于纳米悬浮液中。辅酶Q10纳米悬浮液冻干粉中辅酶Q10的饱和溶解度和体外溶出速率都远高于辅酶Q10原药(P<0.05)；
2、研究了纳米辅酶Q10药代动力学特性结果表明,Tmax(3.3±0.23 h)显著小于辅酶Q10原药的Tmax值(P<0.05),表明辅酶Q10经纳米后在大鼠体内的吸收加快,达峰时间提前。纳米化后药物的Cmax (3.23±0.09μg/mL)显著高于原药的Cmax值(P<0.05),AUC值也大于原药的AUC,表明辅酶Q10经纳米化后在大鼠体内能达到更高的血浆浓度,机体生物利用度也显著增加,以原药(R)作为参比制剂,纳米混悬液(T)的相对生物利用度F为(101±3)%；
3、研究了纳米辅酶Q10体内抗氧化活性结果表明,较原药组和阳性对照组,纳米组显著升高血浆SOD、CAT和GSH-Px活力和肝脏的SOD活力(P<0.05),并明显降低小鼠血浆和肝脏MDA含量(P<0.05),并呈现剂量依赖性,辅酶Q10纳米混悬液对小鼠肝脏和肾脏组织超微结构无任何病理学影响,是安全可靠的新剂型；
4、研究了纳米辅酶Q10透皮能力和抗UVB皮肤老化通过体外皮肤渗透实验研究,表明辅酶Q10纳米混悬液具有优良的透皮能力,稳态渗透率由为13.54±1.10μg·cm-2·h-1,是市售商品的5.46倍。在UVB皮肤光老化实验中,较原药组和美丽加芬阳性对照组,纳米混悬液组显著升高血浆和皮肤中SOD、CAT和GSH-Px活力(P<0.05),并明显降低小鼠血浆和皮肤中MDA含量(P<0.05),提高了皮肤中羟脯氨酸的含量(P<0.05)。
综上所述,辅酶Q10纳米混悬液是辅酶Q10无定形分散体系,载药量高,粒子形态大多为圆球形,分散性良好,粒径约为200±50 nm,具有良好的稳定性。辅酶Q10纳米混悬液能够提高药物溶解性、稳定性和生物利用度,增强了药效、降低不良反应、减少用药剂量等特点。
Coenzyme Q10 is mitochondrial energy source and natural antioxidants in organism, having an important physiological role and clinical value. However, the coenzyme Q10 is a fat-soluble substances, with poor human absorption, low bioavailability and poor stability constituting a bottleneck in its formulation and application.
In recent years, nanosuspension is a new nano-drug delivery system, can increase drug solubility, stability and bioavailability,but also achieve targeted drug delivery, controlled drug release, increase drug stability, reduce side effects, reduce the dosage and so on.
Therefore, the aims of the study is to assess coenzyme Q10 suspension containing nano-medicine system from aspects of quality control and the pharmacodynamic. First of all, investigated preparation for the characterization of nano suspensions the quality of coenzyme Q10; we did pharmacokinetics experimental in rats to investigate in vivo absorption and metabolism; to investigate effect of oral coenzyme Q10 nano-suspension on antioxidative enzyme system,we detect the SOD, CAT, GSH-Px activity and MDA content; carried out in vitro rat skin permeation tests, to investigate the penetration ability of coenzyme Q10 nano-suspension, and the establishment of UVB light skin aging model to study nanosuspension coenzyme Q10 to see skin photoaging ability. Thus, for the clinical application of coenzyme Q10, health products, food and cosmetics fields widely used, open up good prospects. The results of the research topics are as follows:
1、Stability of quality assessment and quality evaluation of coenzyme Q10 and stability of nano-suspension Transmission electron microscopy shows that coenzyme Q10 as spherical particles, diameter of 200±50nm, good dispersion and stability of the system. X-ray diffraction and DSC show amorphous form of Coenzyme Q10 present in the nano-suspension. Nano Suspensions of coenzyme Q10,freeze-dried powder in saturated solubility and dissolution rate of coenzyme Q are much higher than the original drug, nanoparticle size could be reduced, increasing the particle surface area, to further increase its solubility;
2、Pharmacokinetics Tmax (3.3±0.23 h) was significantly smaller than the original drug coenzyme Q10 Tmax values (P<0.05), showed that coenzyme Q10 by the nano-absorption in rats after the accelerated time to peak early. Nanocrystalline drug Cmax (3.23±0.09μg/ mL) was significantly higher than the original drug Cmax values (P<0.05), AUC=78±2.49μg/(mL·h) value is also larger than the original drug AUC values that coenzyme Q10 in rats after by the ultra fine powder to achieve a higher plasma concentration, bioavailability body also increased significantly 1 times the original drug;
3、Antioxidant activity in mouse Compared to ordinary powder group and positive control group had significantly higher ultrafine plasma SOD, CAT and GSH-Px activity and liver SOD activity (P<0.05),and significantly reduced plasma and liver MDA content in mice (P<0.05), dose-dependent manner, coenzyme Q10 on the ultrastructure of mouse liver and kidney without any pathological effects of the new formulation is safe and reliable;
4、UVB Skin aging and penetration ability The studies about skin permeation in vitro showed that coenzyme Q10 nano-suspension has good penetration capacity, steady-state penetration rate of Js was 17.54±1.10, is a commercial product is marketed product 12 times. UVB skin aging in the experiment, compared with ordinary powder Li Jiafen positive control group and U.S.group had significantly higher ultrafine plasma and skin in the SOD, CAT and GSH-Px activity (P<0.05),and significantly reduced in mice MDA content in plasma and skin (P<0.05), increased content of hydroxyproline in the skin.
In summary, the coenzyme Q10 nanosuspension is coenzyme Q10 amorphous dispersion, has high drug loading, and particle shape are mostly spherical.well dispersed system with particle size of about 200±50 nm, and has good stability. Coenzyme Q10 nanosuspension increases drug solubility, stability and bioavailability, and increased efficacy, reduces side effects and the dosage and so on.
引文
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