【作者】 张驰；

【导师】 陈铭；

【作者基本信息】 厦门大学 ， 海洋生物学， 2019， 硕士

【摘要】 本研究介绍了一种新的经皮给药技术,利用蜂海绵骨针与脂质体给药系统的协同作用,可以显著增加亲水性和疏水性大分子药物的经皮吸收。我们经由体外与体内透皮实验,证明了蜂海绵骨针与柔性脂质体可产生协同作用,大幅增加透明质酸的皮肤吸收,尤其是在皮肤深层的吸收。经蜂海绵骨针按摩后,每平方毫米皮肤表面可形成850±125个深度为48.6±13.5微米的微通道,这些微通道会持续存在120小时。在此期间,皮肤通道持续开启,完整的柔性脂质体囊泡会在皮肤深层与表层产生的水合梯度作用力的驱动下,包裹或携带大分子透明质酸,通过变形作用进入皮肤通道,将透明质酸输送到皮肤深层。在体外透皮实验中,骨针与柔性脂质体协同作用16小时后,离体皮肤的透明质酸吸收率为23.2±3.7%,是对照组的19.4±3.1倍(p<0.001),是骨针组的3.4±0.5倍(P<0.01)以及微针+柔性脂质体组的3.6±0.6倍(p<0.01)。而且,蜂海绵骨针与柔性脂质体协同作用的给药方式可以增加所有皮层,尤其是皮肤深层的药物吸收:体外实验中骨针+柔性脂质体组的皮肤深层透明质酸吸收率高达86.8±4.1%。在体内实验中,蜂海绵骨针与柔性脂质体协同作用6小时后,小鼠皮肤的透明质酸吸收率可达8.1%±1.6%,是对照组的7.0±1.4倍(p<0.01),柔性脂质体组的2.3±0.4倍(p<0.05)以及骨针组的2.1土0.4倍(p<0.05)。刺激性实验以及皮肤毒性试验证明了蜂海绵骨针与柔性脂质体联合作用的给药方式仅会对豚鼠皮肤造成轻微刺激,并且会在短时间内恢复正常,因此蜂海绵骨针与柔性脂质体联合作用,可安全有效的增加如透明质酸一样的亲水性大分子药物的经皮吸收。同时,我们也通过体外实验证明了蜂海绵骨针与醇质体协同作用促进环孢素A经皮递送的有效性。经过16小时的体外透皮,蜂海绵骨针与醇质体的组合药物吸收总率为32.98%±0.71%,其中活性表皮层药物吸收率为14.36%±1.44%,占药物吸收总率的43.55%±4.14%,是骨针组的1.53±0.15倍,是醇质体组的2.99±0.28倍。由此可见蜂海绵骨针与醇质体可产生协同作用,增加疏水性药物环孢素A在活性表皮层的皮肤吸收。综上所述,蜂海绵骨针既可以协同柔性脂质体,促进亲水性大分子药物的经皮吸收,也可以与醇质体共同使用,增加疏水性药物的皮肤渗透。这一全新的给药方式不仅克服了固体微针与微针贴片的固有弊端,具有使用灵活,长效促渗的优点。而且与两种技术的单独使用相比,进一步提升了药物的经皮吸收效果,为大分子药物的经皮递送提供了更加安全有效的选择。更多还原

【Abstract】 A new skin drug delivery technology was introduced in this study to increase the skin absorption of hydrophilic and hydrophobic drugs with the the combined use of sponge spicules and liposomes.We demonstrated that the topically combined use of sponge Haliclona sp.spicules(SHS)and flexible liposomes(FL),referred to as SFLS(SHS-Flexible Liposomes combined System),can result in synergy to improve the skin absorption and deposition of Hyaluronic acid(HA),especially in skin deep layers,both in vitro and in vivo.SHS treatment can result in skin micro-channels which are continuous,deep enough(48.6±13.5μm)and available in large quantities(850±125 micro-channels per mm2).These micro-channels gradually closed up in 120 hrs and also allowed the intact vesicles of flexible liposomes as well as vesicle-bound or vesicle-encapsulated HA to penetrate into the skin-deep layers under the driving force of transdermal osmotic gradients.Specifically,SFLS topical application enhanced the penetration of FITC-HA(MW:250 kDa)into porcine skin in vitro up to 23.2±3.7%,which is 19.4:1±3.1-fold higher(p<0.001)than Phosphate Buffered Saline(PBS)group,3.4±0.5-fold higher(p<0.01)than SHS group and 3.6±0.6-fold higher(p<0.01)than that from the combined use of Dermaroller and flexible liposomes.Moreover,SFLS can lead to significant enhanced skin deposition of HA in all skin layers,especially in skin deep layers:up to 86.8 ± 4.1%of HA skin absorbed was accumulated in deep skin layers.The effectiveness of SFLS topical application was also confirmed in vivo by using BALB/c mice.The HA percutaneous penetration induced by SFLS after 6h non-occlusive topical application in vivo(8.1 ± 1.6%)was dramatically increased,which is 7.0±1.4-fold higher(p<0.01)than PBS control,2.3±0.4-fold higher(p<0.05)than FL group,and 2.1 ±0.4-fold higher(p<0.05)than SHS group.In addition,the skin irritation and toxicity study showed that the SFLS treatment may cause very minimal redness and the skin can get recovery in short time.In sum,the combined use of SHS and FL(SFLS)offers a promising strategy to safely and effectively improve skin delivery of hydrophilic biomacromolecules such as HA.Meanwhile,we also demonstrated that the topically combined use of SHS and ethosomes can result in synergy to improve the skin absorption and deposition of Cyclosporine A(CSA).The combined use of SHS and ethosomes enhanced the penetration of CSA into porcine skin in vitro up to 32.98±0.71%,14.36± 1.44%of CSA was accumulated in viable epidermis layers,which accounts for 43.55±4.14%of total skin absorption of CSA.It is 1.53±0.15-fo]d higher than SHS group,and 2.99±0.28-fold higher than ethosomes group.Above all,SHS can be used in conjunction with both flexible liposomes and ethosomes to enhance the absorption of HA and CSA.Compared with traditional micro needles and microneedle patch,It provides a more convenient,effective and safe way for skin drug delivery.更多还原