【摘要】：多肽類(lèi)藥物口服給藥的研究一直是國(guó)內(nèi)外藥劑學(xué)研究者熱衷的領(lǐng)域,但這類(lèi)藥物由于自身理化性質(zhì)等特點(diǎn),口服生物利用度低。這其中研究者尤為關(guān)注的是胰島素。胰島素是臨床上治療I型糖尿病的一線(xiàn)藥物,但存在的突出問(wèn)題是藥物半衰期短,需長(zhǎng)期頻繁地注射給藥。而胰島素口服給藥是一種患者順應(yīng)性好、服用方便的給藥方式,符合內(nèi)源胰島素的分泌模式。然而,胰島素直接口服給藥時(shí)遭遇胃腸道酶降解以及膜透過(guò)性差等諸多障礙。生物黏附性聚合物納米�？梢酝ㄟ^(guò)延長(zhǎng)胃腸道藥物滯留時(shí)間和減少胃腸道中消化酶對(duì)胰島素的降解等方式,有效改善胰島素口服生物利用度。 本研究將生物黏附性聚合物透明質(zhì)酸與L-半胱氨酸通過(guò)酰胺鍵連接制備了巰基化透明質(zhì)酸(HA-Cys),其游離巰基含量為(225.7±4.4)μmol·g-1,核磁共振以及紅外圖譜確證了其結(jié)構(gòu)。體外黏附性實(shí)驗(yàn)表明聚合物黏附性質(zhì)良好。 以HA-Cys為載體,采用超聲乳化法,制備了載胰島素的巰基化透明質(zhì)酸納米粒(Ins-HA-Cys-NPs)。通過(guò)單因素考察,確定了對(duì)Ins-HA-Cys-NPs影響較大的因素為油相、水相體積比,HA-Cys用量以及Ins用量,并通過(guò)星點(diǎn)設(shè)計(jì)-效應(yīng)面優(yōu)化法,以包封率、載藥量、粒徑以及多分散指數(shù)為評(píng)價(jià)指標(biāo),優(yōu)化了納米粒的制備工藝。最終得到的最優(yōu)處方為水相、油相體積比為1：4.12,HA-Cys用量為25.95mg, Ins用量為8.04mg。通過(guò)激光粒度分析儀測(cè)定納米粒粒徑和分布,平均粒徑(PS)為(178.5±0.75)nm,多分散系數(shù)(PDI)為0.214±0.01,Zeta電位(ZP)為(-38.47±0.46)mV；反相高效液相法測(cè)定納米粒包封率和載藥量,包封率(EE)為(48.85±0.66)%,載藥量(DL)為(4.79±0.13)%。進(jìn)行了納米粒的形態(tài)觀察、黏附性質(zhì)、體外釋放與抗酶降解特性的研究,評(píng)價(jià)了胰島素巰基化透明質(zhì)酸納米粒的大鼠口服給藥的降血糖效果。實(shí)驗(yàn)結(jié)果表明,所制得的Ins-HA-Cys-NPs外觀圓整,且分布均一；制備的Ins-HA-Cys-NPs具有良好的體外黏附特性,黏蛋白與Ins-HA-Cys-NPs納米�；鞈乙夯旌虾骦eta電位由(-17.32±0.38)mV下降到(-36.06±0.62)mV；與胰島素溶液相比,Ins-HA-NPs及Ins-HA-Cys-NPs藥物釋放時(shí)間均延長(zhǎng)；以胃蛋白酶、胰蛋白酶為代表,研究Ins-HA-Cys-NPs的抗酶解作用,說(shuō)明Ins-HA-Cys-NPs具有較好的抗酶降解作用；藥效學(xué)實(shí)驗(yàn)中,糖尿病大鼠口服Ins-HA-Cys-NPs后1-12h中,顯示出平緩的降血糖效果,相對(duì)生物利用度為6.9%。
[Abstract]:The study of oral administration of polypeptide drugs has always been a hot field of pharmacists at home and abroad, but the oral bioavailability of these drugs is low due to their physical and chemical properties. Of these, researchers are particularly concerned about insulin. Insulin is a first-line drug in the treatment of type I diabetes, but the outstanding problem is that the half life of the drug is short and it needs to be injected frequently for a long time. The insulin oral administration is a kind of patient's compliance, the convenient administration way, conforms to the endogenous insulin secretion pattern. However, insulin directly administered orally encountered many obstacles, such as gastrointestinal enzyme degradation and poor membrane permeability. Bioadhesive polymer nanoparticles can effectively improve the oral bioavailability of insulin by prolonging the drug retention time in the gastrointestinal tract and reducing the degradation of insulin by digestive enzymes in the gastrointestinal tract. In this study, mercaptohyaluronic acid (HA-Cys) was prepared by linking the bioadhesive hyaluronic acid with L-cysteine by amide bond. Its free thiol content was (225.7 鹵4. 4) 渭 mol g ~ (-1). Its structure was confirmed by NMR and IR spectra. In vitro adhesion test showed that the polymer adhesion was good. The sulfated hyaluronic acid nanoparticles (Ins-HA-Cys-NPs) loaded with insulin were prepared by phacoemulsification with HA-Cys as carrier. By single factor investigation, the oil phase, the volume ratio of water phase HA-Cys and the amount of Ins were determined, and the encapsulation efficiency, drug loading, particle size and polydispersity index were used as the evaluation indexes by the method of star design-effect surface optimization. The preparation process of nanoparticles was optimized. The optimum formulation was water phase, the volume ratio of oil phase was 1: 4.12, HA-Cys was 25.95 mg, and the dosage of Ins was 8.04 mg 路mol ~ (-1) 路min ~ (-1) 路min ~ (-1). The particle size and distribution of nanoparticles were measured by laser particle size analyzer. The average particle size (PS) was (178.5 鹵0.75) nm, and the polydispersity coefficient (PDI) was 0.214 鹵0.01U Zeta potential (ZP) was (-38.47 鹵0.46) MV. The encapsulation efficiency and drug loading capacity of nanoparticles were determined by reversed-phase high performance liquid chromatography (RP-HPLC), the entrapment efficiency (EE) was (48.85 鹵0.66), and the drug loading (DL) was (4.79 鹵0.13) mm. The effects of oral administration of insulin mercaptohyaluronic acid nanoparticles on hypoglycemia in rats were evaluated by means of morphological observation, adhesion properties, release in vitro and anti-enzymatic degradation of nanoparticles. The results showed that the prepared Ins-HA-Cys-NPs had a round appearance and uniform distribution, and the prepared Ins-HA-Cys-NPs had a good adhesion property in vitro, and the Zeta potential decreased from (-17.32 鹵0.38) MV to (-36.06 鹵0.62) MV after the mixture of mucin and Ins-HA-Cys-NPs nanoparticles suspension, and the Zeta potential decreased from (-17.32 鹵0.38) MV to (-36.06 鹵0.62) MV. Compared with insulin solution, the release time of Ins-HA-NPs and Ins-HA-Cys-NPs was prolonged. The antienzymatic hydrolysis of Ins-HA-Cys-NPs was studied by using pepsin and trypsin as the representative. Diabetic rats showed a gentle hypoglycemic effect in 1-12 hours after oral administration of Ins-HA-Cys-NPs, and the relative bioavailability was 6. 9%.
【學(xué)位授予單位】：廈門(mén)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R943

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