단백질 약물 전달을 위한 수화젤, 융합 단백질 그리고 전기방사 마이크로 입자의 개발에 관한 연구

Abstract

Recent advances in pharmaceutical biotechnology have produced an increasing number of new drug candidates such as novel peptide and protein drugs as well as nucleic acid based drugs for gene therapy. However, efficient delivery of these biopharmaceuticals has been considered to be a major challenge because their intrinsic properties lead to poor physical and chemical stability in the body and limited membrane permeability, resulting in low bioavailability and low efficacy. In addition to the stability and permeability issues, targeted delivery of biopharmaceuticals to disease sites is important to maximize efficacy and minimize toxicity by reducing dose and frequency. In this study, various delivery systems including hydrogels, recombinant fusion technology and electrospun micro-particles have been studied to develop optimized dosage forms for efficient protein drug delivery. Thermo-reversible hydrogel based on low molecular weight methylcellulose was prepared and tested to improve protein drug stability and achieve a sustained release pattern. It was found that thermo-reversible hydrogel could reduce the number of injection and deliver a protein drug while maintaining its efficiency. The protein transduction domain such as Tat peptide was fused in a recombinant method to a protein, Heat Shock Protein 27 (Hsp27), which is known to be preventive against ischemic heart diseases. It showed that Tat-Hsp27 fusion protein increasing the cardiac stem cell viability by apoptosis pathway. Lastly, electrospray technology that could adjust size and shape of high molecule polymer containing proteins by using high voltage was applied to produce core/shell micro-particle. Protein loaded core/shell micro-particle was successfully fabricated and its core/shell structure was confirmed. Although further intensive studies are needed, three delivery systems mentioned above provide promising results for protein drug delivery.|단백질 약물은 부작용이 적은 점과 효능에서 뛰어난 평가를 받지만 구조 자체의 안정성과 목표 지점까지의 접근성, 그리고 잦은 투여 횟수에 대한 단점이 지적되어 왔다. 이러한 단점을 극복하기 위해 많은 약물 전달 시스템이 연구되어 왔으며 실제로 많은 개발이 이루어졌다. 이 연구에서는 이러한 추세에 발맞추어 단백질 약물 전달을 위한 다양한 접근 방식에 대해 폭넓게 살펴보았다. 단백질의 구조적 안정성을 높이고 투여 횟수를 줄이기 위하여 온도 감응성 젤을 이용, 하이드로젤의 약물 용출 경향을 확인한 뒤 동물 모델에 적용하여 단백질 약물 전달로서의 하이드로젤의 효능을 평가하였다. 또한 허혈성 심근질환에 대하여 효능이 있는 열충격단백질 (Hsp27)의 단백질의 세포 투과성을 높이기 위하여 단백질 전달 물질 (PTD)을 부착한 융합 단백질을 생산하여 동물 심장 줄기 세포에 미치는 영향과 그 메커니즘을 분석하였다. 마지막으로, 고전압 전류를 이용해 고분자 화합물의 크기와 모양을 조절할 수 있는 전기방사를 이용, 단백질 약물을 봉입한 코어셸 구조의 마이크로 크기의 입자를 생산하기 위한 조건을 확립하였다. 확립한 조건 하에 방사된 입자가 코어셸 구조를 유지하고 있는지 여부를 평가하였으며 추후 동물 모델 실험에 대비하고자 약물 단백질 용출 실험을 진행하였다. 본 연구를 통하여 단백질 약물의 안정성을 유지하면서 투여 빈도를 낮출 수 있는 방법과 단백질 약물의 구조적 개선을 통해 그 단백질의 효능 및 효율을 높일 수 있는 방법을 검증하였다; Recent advances in pharmaceutical biotechnology have produced an increasing number of new drug candidates such as novel peptide and protein drugs as well as nucleic acid based drugs for gene therapy. However, efficient delivery of these biopharmaceuticals has been considered to be a major challenge because their intrinsic properties lead to poor physical and chemical stability in the body and limited membrane permeability, resulting in low bioavailability and low efficacy. In addition to the stability and permeability issues, targeted delivery of biopharmaceuticals to disease sites is important to maximize efficacy and minimize toxicity by reducing dose and frequency. In this study, various delivery systems including hydrogels, recombinant fusion technology and electrospun micro-particles have been studied to develop optimized dosage forms for efficient protein drug delivery. Thermo-reversible hydrogel based on low molecular weight methylcellulose was prepared and tested to improve protein drug stability and achieve a sustained release pattern. It was found that thermo-reversible hydrogel could reduce the number of injection and deliver a protein drug while maintaining its efficiency. The protein transduction domain such as Tat peptide was fused in a recombinant method to a protein, Heat Shock Protein 27 (Hsp27), which is known to be preventive against ischemic heart diseases. It showed that Tat-Hsp27 fusion protein increasing the cardiac stem cell viability by apoptosis pathway. Lastly, electrospray technology that could adjust size and shape of high molecule polymer containing proteins by using high voltage was applied to produce core/shell micro-particle. Protein loaded core/shell micro-particle was successfully fabricated and its core/shell structure was confirmed. Although further intensive studies are needed, three delivery systems mentioned above provide promising results for protein drug delivery.