[논문]Succinylated Pullulan Acetate Microspheres for Protein Delivery

Woo, Young-Rong; Seo, Seog-Jin; Na, Kun

doi:10.4333/kps.2011.41.6.323

Succinylated Pullulan Acetate Microspheres for Protein Delivery 원문보기

Journal of pharmaceutical investigation, v.41 no.6, 2011년, pp.323 - 329

Woo, Young-Rong (Department of Biotechnology, The Catholic University of Korea) , Seo, Seog-Jin (Department of Biotechnology, The Catholic University of Korea) , Na, Kun (Department of Biotechnology, The Catholic University of Korea)

Abstract ▼ AI-Helper

In order to develop new protein carrier replacing poly(DL-lactic acid-co-glycolic acid) (PLGA) microspheres, succinylated pullulan acetate (SPA) was investigated to fabricate a long term protein delivery carrier. SPA microspheres loaded with lysozyme (Lys) as a model protein drug were prepared by a water/oil/water (W/O/W) double emulsion method. An acidity test of SPA copolymers after hydrolysis was performed to estimate the change of protein stability during releasing proteins from the microspheres. There was no pH change of SPA copolymers, but pH of PLGA polymers after hydrolysis was significantly decreased to around pH 2, indicating that the long-term stability of proteins released from SPA microspheres can be guaranteed. Loading efficiency of proteins into SPA microspheres was three times higher than those into conventional PLGA microspheres, indication of inducing stronger charge interaction between proteins and succinyl groups in SPA microspheres. Although initial burst behaviors were monitored in Lys-loaded SPA microspheres due to relatively strong hydrophilic succinyl segments in SPA microspheres, initial burst issues would be circumvented if the ratio of charge density of succinyl moieties and hydrophobic acetate groups is harmonically controlled. Therefore, in this study, a new attempt of protein delivery system was made and functional SPA was successfully confirmed as a new protein carrier.

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문제 정의

, 2005). These works were attributed to high stability of protein drugs from acylation when degrading PLGA. Another group reported on using the system with natural polymers to protect protein drugs in acidic environment of PLGA microsphere(Lee et al.

제안 방법

In this study, we synthesized succinylated pullulan acetate (SPA; Figure 1(c)) as a protein delivery depot using negatively charged succinyl anhydride and hydrophobic acetic anhydride, and confirm the resultants by ¹H nuclear magnetic resonance (NMR). Because the carboxyl groups in SPA have a pKa around pH 4.
Shapes of PLGA, SPA1 and 2 microspheres were similarly regular and spherical as visualized by FE-SEM photography and they were not affected by the succinyl group contents. Lysloaded SPA2 microspheres were expected to be slightly smaller and more compact than Lys-loaded PLGA and SPA1 microspheres, but the significant results concerning size distribution among them were not analyzed in this study.

이론/모형

, 2010). Here, Lys as a model drug was encapsulated into SPA microspheres via a water/ organic/water (W/O/W) double-emulsion method. The proteinloaded SPA microspheres were expected to reduce initial burst of protein drugs and to sustain long-term release of proteins.
SPA microspheres were prepared via a double-emulsion-solvent evaporation technique (W/O/W, water-in-oil-in-water). Five mg of Lys dissolved in 0.

성능/효과

In conclusion, we successfully synthesized SPA copolymers composed of negatively charged succinyl groups and hydrophobic acetyl groups grafted to pullulan in effort to fabricate microspheres for the future development of new protein drug carriers. SPA microspheres evidenced that the long-term stability of proteins in the microspheres were maintained as a result of unchanged pH unlike PLGA microspheres.
The microparticles were characterized by their morphological structure and their size by the field emission scanning electron microscopy (FE-SEM). The protein encapsulation efficiency and release profiles of Lys in vitro were examined, and its stability was also assessed by a pH test compared to those in conventional PLGA microspheres.
3%, respectively, whereas those in SPA microsphers were about three times higher than those in PLGA microspheres. These results indicated that SPA copolymers can induce not only hydrophobic forces between Lys and acetyl groups but also charge interaction between protein drugs and succinyl groups in SPA microspheres. In contrast, PLGA microspheres have only hydrophobic forces between proteins and PLGA when proteins were loaded into the microspheres.
There were no pH changes of SPA1 (Figure 4b) and SPA2 (Figure 4c) copolymers between before and after hydrolysis, but pH of PLGA polymers (Figure 4a) after hydrolysis was significantly decreased to around pH 2 as shown in Figure 4. These results indicated that proteins encapsulated in SPA microspheres are stable, and it is possible to perform long-term release of therapeutic protein drugs in SPA microspheres.
Although SPA microspheres were endowed with charge functions for stably ionic interaction between Lys and the microspheres, initial burst behaviors of Lys from SPA microspheres could not be avoided by thereby higher hydrophilicity. These results suggested that relatively hydrophobic forces were weaker, the more highly hydrophilic succinyl segments despite higher loading efficiencies of proteins into the microspheres by stronger charge interaction, thus resulting in releasing out of almost all the proteins from the microspheres. Therefore, if the ratio of charge density and hydrophobic forces are harmonically controlled, the initial burst problems of the microspheres can be solved.

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