1 1，■l 」

Ap미led Chemistly,

Vol 13. No. 2, October 2009. 181. 184

cisPlatin이 봉입된 수용성 Ca『boxymathyl Chito$ an m icel le의 체조

뵤뵤프 · 정현 · 김원석 · 최창용� 정영일� 장미경 · 나재운’ 순천대학교 공과대학 고분자공학과

(E. mail : jwnah@ sunchon.ac.kr ’)

P『epa『ati。n of ci$ Platin- IncO『po『ated Ca『b。xymethylWate 『－＄이Uble Chit0San MICelle$

拉브峴그盤뜨끄흐쯔 · Hyun Jung · WOn- Seok Kim· Changyoung Choi. Young. 11 Jeong · ·Mi. Kyeong Jang · Jae- Woon Nah ’

Department of Polymer Science and Engineering, Sunchon National University,

Suncheon, Jeonnam 540- 742, Korea

(E. mail : jwnah@ sunchon. ac. kr)

ABSTRACT

In this Study, we prepared micelles based on the formation of an

ion- complex between carboxymethyl water- soluble chitosan (CMCh)

and cisPlatin. Cisplatin (cis ’dichlorodiammineplatinum (11) : CDDP) 15

one of the most widely used antitumor drugs in the treatment of a

variety of solid tumors. CDDP loaded CMCh (Cis- CMCh) micelles were

prepared by ion- complex and investigated by IH- NMR, FT- IR

(Fourier transform infrared) , TEM (scanning 이eCtron microscopy) ,

DLS (dynamic light scattering) , release behavior and cells cytotoxicity

measurement. The average diameter, drug content, and drug foading

efficiency of Cis- CMCs- NP were 3OO.snm, 7. 1% (w/ w) , and 76. 0% ,

respectively.

1. IntroduCtion

Chitosan a copolymer of glucosamine and N- acetyglucosamine units hnked by

1 . 4 glucosidic bonds, is obtained by N- deacetylation of chitin, which is the

second most naturally occurring biopolymer (after cellulose) 【2] . It has been

found a number of appllcations in drug delivery including that of adsorption

enhancer of hydrophilic macromolecular drugs 【3] . Cisplati미

cis- diamineplatinum- (11) dichloride (CDDP) , 15 a chemotherapeutic drug used for

the treatment of neuroblastomas, and testicular, bladder, ovarian, and other solid

cancers [ 51. However, it 15 well known that its efficiency 15 restriCted by serious

side effects, such as the emergence of intrinsic and acquired resistance, acute

nephrotoxicity and chronic neurotoxicity [ 6, 7] . In this study, we prepared

nanoparticles based on the formation of an ion complex between carboxymethyl

chitosan (CMCh) and cisplatin. The physicochemical properties of the

cisplatin- incorporated nanoparticles of CMCh, as wen as their cisplatin release

18l

1 82 남정표 · 정 현 · 김원석 · 최창용 · 정 영 일 · 장미경 · 나재운

behavior from

investlgated.

the nanoparticles and HEK 293T cells cytotoxicity, were

2. Experimental

Preparation of cisPlitin- incorporated CMCh micenes (Cis- CMCh)

50 mg of o- CMCh and N, O- CMCh was dissolved in 5 ml of deionized water

(0. 1 %, w/ v) without addition of acetic acid, respectively. Also, 50 mg of

N- CMCh was dissolved in 5 ml of deionized water (0. 1 %, w/ v) . The reaction

mixtures were stirred for 30min at room temperature. 10 mg of cisplatin powder

was added to solution of 0 and N, O- CMCh, respectively. Also, 5, 10 and 20 mg

of cisPlat1n powder was added to soluti0n of N- CMCh. The mlxtures were stirred

gently for 3 days at room temperature. Subsequently the resulting solution was

dialyzed against deionized water for 6 h with stirring to remove unwanted trace

elements. The dialyzed solution was lyophihzed for 3 days or anaiyzed.

.（〕，「 ．》．〈〕， , . t驢 ‘ 》

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* '’톨‘’륵�c。 ，＊눕＊ ’. ’ 톨읨뺨껍C터＊ 邇노 ‘

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."N . /n ．·，t ,^

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:：二 "0 0 · 1』맥

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’ 1 ’ . � 칙 茗 날 r’ 1 ’ 냥 ’.

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q: , , "q콥떠노

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*.’x:(, ．〔 꽈（커신 ·C esO O . d

. ，구끄”철c

.．ㅐ ,

嚼／" b

흼’1’x '”구？驪，.,，�＿_ , , -‘택

( 》 ( , ’ 《 - ．·

∥쇼／ (，卜｀ ‘》《】九1 《 h

！탬끄1

Figure 1. Schematic illustrations of polyeleCtrolyte complex formation

Cis- NCMCh (a) and Cis- OCMCh (b) .

Transmission electron microscope (TEM) observation

Cis- CMCh nanoparticles morphology was examined with

FX- H transmission electron microscope (TEM) . A drop

JEOL JEM- 2000

of Cis. CMCh

nanoparticles suspended in o,01%

carbon film coated on a copper grid

of ph0sph0tungstic acid were Placed 0n a

for TEM.

CisPlatin release from Cis- CMCh nanoparticles

3 mg of the Cis- CMCh nanoparticles was resuspended int0 2 ml of

phosphate- buffered saline (PBS, pH 7. 4) and transferred to a dialysis tube

(MWCO: 12, 000 9/ mol) . The dialysis tube was put into a 50 ml bottle with 40 ml

of PBS (O.I M, pH 7. 4) . Release test was performed at 370C with stirring rate of

10Orpm. The whole released medium was exchanged with fresh medium at

predetermined time to maintain the sink condition. Released drug was measured at

응용화학， 제 13 권 제 2 호， 2009

4. -

Cisnlatinol 봉입된 수용성 Carboxymethyl Chitosan rm*celle의 제조

302 nm 【1 6] with UV

calculated as acumulative

spectr0photometer (UV- 1200, ShimadzuCO. ,Ltd. Japan)

value.

3. Result and discussion

In this study, cisPlatin- incorporated micenes were prepared through the

ion- complexes between cisplatin and CMCh, as illustrated in Figure l, The

micelles were ion comPlexes formed by the simple mixing of cisplatin and CMCh.

The morphology of Cis- CMCh micenes was shown Figure 2. As shown in Figure

2, Cis- CMCh (Cis- NCMCh s) micenes had spherical shapes with particle size of

100 . 300 nm. The size and dlstribution of Cis- CMCh (Cis- NCMCh s) 5 micenes

in aqueous environmentS were measured by dynamic hght scattering. Cis- NCMChs

micelles formed 300 nm sized unimodal micelles with fairly narrow distributions.

。O！실．쇼．눕．

.-0

．샅ㄷ·

．一

Figure 2. Particle distribution and TEM image of Cis- NCMChsmicene

Figure 3 showed

SIZe

the cisPlatin release

Figure 3(a) , cisPlatin was continuously

from Cis- CMCh mlcenes, As shown in

released from micelles over 20 h at all

formulations. Higher cisplatin

from micelles. In the case

higher than 80 % for 12 h

contents induced decreased

of Cis. NCMCh s, released

whilP Cig- NCMCh 10 and

release

fraction

rate of cisplatin

of cisplatin was

Cis , NCMCh

relatively zero- order kinetics. Release

20 at 20 h was about 50 % and 40 %,

released in the case of Cis- NCMCh

Cis- NCMCh 10 and Cis- OCMCh 1 0.

fraCtion of Cis- NCMCh 10 and

20 showed

Ci s - NCMCh

drug was

Compare

released

O

>,

0

t

/.

’

11

1

’

respectively, while alm0St

5. Figure 3 (b) , graph

of the

꺼Za S

1n

fraction of cisPlatin was higher than 80 %

the

for

case of Cis- OCMCh

20 h while Cis- NCMCh

Applied Chemistry, Vof. 13, No. 2, 2009

1 84 남정표 · 정 현 · 김원석 · 최창용 · 정영일 · 장미경 · 나재운

畺 ，。·긍

글

쟎

l 1 5

T - m e lh )

畺궁

占낌 · O

첩 4 。軾

寒짊섬

콜금

C ］· _ O 「

T -m e 4h l

Figure 3. The release of cisPlatin from Cis- NCMCh (5, IOand20) (a) and

Cis- OCMCh1O (b) .

4. Conclusion

CisPlatin- incorporated nanoparticles were prepared by simple mixing of cisp- atin

and CMCh. Nanoparticles can be prepared by ion comPlex formation between

cisplatln and CMCh. At the observation of transmission electron microscopy

(TEM) , cisplatin- incorporated CMCh nanoparticles have spherical shapes with

diameter of about 100 nm , 300 nm. At drug release study, cisplatin release rate

was decreased by increased feeding amount of cisplatin. Nanoparticles from

OCMCh showed faster cisplatin release properties than that of NCMCh.

ReferenCe

I RAA, Muzzarelli, CarbohydrP0lym, 29, 309 (l996) .

2. P. Artursson, T. Lindmark, 5. 5. Davis and L. nlum, Effect of chitosan on the permeabihty of

monoIayers of intestinal epithelial-cells (caco-2). PharmaceuticaIResearch, ∥， 1358-1361 (l994).

3· P. J. Loehrer and L. H. Einhorn, CisPlatin, Ann.IntMed, 100, 704-713 (1984).

4. M. Kartalou and J. M. Essigmann, Mechanism of resistance to cisPlatin. Mutation

Research, 478 ,23- 43 (2001) .

5. V. Ponzani, F. Bress0Ile, 1. J. Haug, M. Galtier and J. P. Blayac, CibPlatin-induced renal toxicity

and toxicity-m。dulating strategies:Review. cancerchemotheraPyandPharmac。fogy, 朽， l- 9 (l "4) .

6. D.G. Kim, C.Y. Choi, M.K. Jang and J.W. Nah, Al- -trans retinoic acid associated low molecular

weight water soluble chitosan nanoParticles based on ion comPlex. MacromoI.Res. 14, 66-72

(2006) .

응용화학， 제 l3 권 제 2 호， 2D09