유전자 전달체로서 히스티딘이 결합된 저분자량 수용성 키토산의 제조와 특성

박준규; 김동곤; 최창용; 장미경; 나재운; Jun-Kyu Park; Dong-gon Kim; Changyong Choi; Mi-Kyeong Jang; Jae-Woon Nah

Journal of the Korean Industrial and Engineering Chemistry, Vol.18, No.6, 607-611, December, 2007

Export Citation

유전자 전달체로서 히스티딘이 결합된 저분자량 수용성 키토산의 제조와 특성

Preparation and Characterization of the Histidine-graft-Low Molecular Weight Water-Soluble Chitosan as a Gene Carrier

박준규, 김동곤, 최창용 , 장미경 ^†, 나재운 ^†

순천대학교 공과대학 신소재응용공학부 고분자공학전공

Jun-Kyu Park, Dong-gon Kim, Changyong Choi , Mi-Kyeong Jang^†, and Jae-Woon Nah^†

Department of Polymer Science and Engineering, Sunchon National University, Jeonnam 540-742, Korea

E-mail:,

초록

양이온성 천연고분자 물질로서 유전자와 효과적으로 복합체를 형성할 수 있는 저분자량 수용성 키토산(LMWSC)에 유전자 발현 효율을 높이기 위해 histidine을 컨쥬게이션 시킨 유전자 전달체를 제조하였다. 히스티딘-LMWSC의 제조는 무수프탈 산으로 LMWSC의 아민기를 보호한 후 히스티딘과 결합한 다음 무수프탈산을 제거하는 반응으로 제조하였다. 제조한 유전자 전달체의 물리화학적 특성은 FT-IR과 NMR 스펙트럼을 통하여 확인하였고, 전기영동 결과로부터 본 실험에서 제조된 유전자 전달체와 DNA가 복합체를 형성하였다는 것을 알 수 있었다. 또한 유전자 전달체의 입자크기는 DLS를 이용하여 측정한 결과 히스티딘-LMWSC-DNA 복합체입자의 크기는 100∼200 nm임을 확인하였다. 이로부터 히스티딘을 함유하는 LMWSC가 유전자 전달체로서 사용될 수 있음을 확인하였다.

To improve transfection efficiency, we prepared histidine-low molecular weight water-soluble chitosan (LMWSC) having the potential to form complex with DNA as a cationic polymer. Histidine-LMWSC was synthesized by the esterification reaction and removing phthaloyl group. The histidine-LMWSC was characterized using FT-IR, 1H NMR spectra. Histidine-LMWSC was complexed with plasmid DNA (pDNA) in various polymer/DNA (N/P) weight ratios, and the complex was identified using gel retardation assay. The particle sizes of the hisitidine-LMWSC/DNA complexes were measured on a DLS instrument by fixing the histidine-LMWSC/DNA weight ratio of 10/1. Owing to the utilization of a large excess amount of cationic LMWSC against anionic DNA, the particle size of histidine-LMWSC/DNA complexes was in the range of 100∼200 nm. Therefore, histidine-LMWSC will be useful in the development of gene carriers.

Keywords:LMWSC;histidine;nanoparticle;gene therapy

[References]

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Fernandez-Urrusuno R, Calvo P, Remunan-Lopez C, Vila-Jato JL, Alonso MJ, Pharm. Res., 16, 1576 (1999)
Kotez AF, de Leeuw BJ, Lueben HL, deBoer AG, Verhoef JC, Junginger HE, Int. J. Pharm., 159, 243 (1997)
Jang MK, Jeong YI, Cho CS, Yang SH, Kang YE, Nah JW, Bull. Korean Chem. Soc., 23, 914 (2002)
Hirano S, Inui H, Koski H, Uno Y, Toda T, Chitin and chitosan: Ecologically bioactive polymer, in biotechnology and bioactive polymers, Gebrlrtin, C. and Carraher, C., eds. New York: Plenum Press, 43 (1994)
Son S, Chae SY, Choi C, Kim MY, Ngugen VG, Jang MK, Nah JW, Kweon JK, Macromol. Res., 12(6), 573 (2004)
Kim CH, Park HS, Gin YJ, Son YS, Lim SH, Choi YJ, Park KS, Park CW, Macromol. Res., 12(4), 367 (2004)
Lee KY, Macromol. Res., 13(6), 542 (2005)
Lee M, Nah JW, Kwon Y, Koh JJ, Ko KS, Kim SW, Pharm. Res., 18, 427 (2001)
Akinc A, Thomas M, Klibanov AM, Langer R, J. Gene. Med., 7, 657 (2005)
Jones TA, Gumbleton M, Duncan R, Adv. Drug Deliv. Rev., 55 (2003)
Kichler A, Leborgne C, Coeytaux E, Danos O, J. Gene. Med., 3, 135 (2001)
Patchornic A, Berger A, Katchalski E, J. Am. Chem. Soc., 79, 5227 (1957)
Boussif O, Lezoualc’h F, Zanta MA, Mergny MD, Scherman D, Demeneix B, Behr JP, Proc. Batl. Acad. Sci. U. S. A., 92, 823 (1995)
Neu M, Fischer D, Kissel T, J. Gene. Med., 7, 992 (2005)
Neu M, Fischer D, Kissel T, J. Control. Release, 78, 273 (2002)

[Related Articles]

[1] Hirano S, Polym. Int., 48, 732 (1999)

[2] Fernandez-Urrusuno R, Calvo P, Remunan-Lopez C, Vila-Jato JL, Alonso MJ, Pharm. Res., 16, 1576 (1999)

[3] Kotez AF, de Leeuw BJ, Lueben HL, deBoer AG, Verhoef JC, Junginger HE, Int. J. Pharm., 159, 243 (1997)

[4] Jang MK, Jeong YI, Cho CS, Yang SH, Kang YE, Nah JW, Bull. Korean Chem. Soc., 23, 914 (2002)

[5] Hirano S, Inui H, Koski H, Uno Y, Toda T, Chitin and chitosan: Ecologically bioactive polymer, in biotechnology and bioactive polymers, Gebrlrtin, C. and Carraher, C., eds. New York: Plenum Press, 43 (1994)

[6] Son S, Chae SY, Choi C, Kim MY, Ngugen VG, Jang MK, Nah JW, Kweon JK, Macromol. Res., 12(6), 573 (2004)

[7] Kim CH, Park HS, Gin YJ, Son YS, Lim SH, Choi YJ, Park KS, Park CW, Macromol. Res., 12(4), 367 (2004)

[8] Lee KY, Macromol. Res., 13(6), 542 (2005)

[9] Lee M, Nah JW, Kwon Y, Koh JJ, Ko KS, Kim SW, Pharm. Res., 18, 427 (2001)

[10] Akinc A, Thomas M, Klibanov AM, Langer R, J. Gene. Med., 7, 657 (2005)

[11] Jones TA, Gumbleton M, Duncan R, Adv. Drug Deliv. Rev., 55 (2003)

[12] Kichler A, Leborgne C, Coeytaux E, Danos O, J. Gene. Med., 3, 135 (2001)

[13] Patchornic A, Berger A, Katchalski E, J. Am. Chem. Soc., 79, 5227 (1957)

[14] Boussif O, Lezoualc’h F, Zanta MA, Mergny MD, Scherman D, Demeneix B, Behr JP, Proc. Batl. Acad. Sci. U. S. A., 92, 823 (1995)

[15] Neu M, Fischer D, Kissel T, J. Gene. Med., 7, 992 (2005)

[16] Neu M, Fischer D, Kissel T, J. Control. Release, 78, 273 (2002)