GVHD: PGS.

TS L Th Hng Nhan

Mc lc
H NANO DN TRUYN THUC CHO NO .......................................................................... 2
1 Gii thiu ................................................................................................................................. 2
2 Cu to hng ro mu no ........................................................................................................ 3
2.1 Lch s : ............................................................................................................................. 3
2.2 Cu to ............................................................................................................................... 4
3 Kim sot thi gian gii phng thuc ...................................................................................... 5
3.1 Gii phng thuc dng Extended-Release formulation ..................................................... 6
3.2 Gii phng thuc dng Sustained-Release formulation .................................................... 7
3.3 Gii phng thuc dng Pulsatile-Release formulation ...................................................... 7
4 Kim sot v tr gii phng thuc theo c ch ch ng .......................................................... 8
4.1. Con ng dn truyn absorptive-mediated transcytosis ............................................... 10
4.2 Con ng dn truyn transporter-mediated transcytosis ............................................... 18
4.3 Con ng dn truyn receptor-mediated transcytosis ................................................... 23
4.4 H dn tryn thuc nhy cm mi trng ....................................................................... 28
5 Kim sot v tr gii phng thuc theo c ch th ng (hiu ng EPR) .............................. 31
6 V d v thit k mt h dn truyn thuc vo no in hnh ................................................ 34
6.1 Khi nim Dendrimer ...................................................................................................... 34
6.2 t vn . ....................................................................................................................... 35
6.3 Phng thc tng hp v c ch dn truyn ................................................................... 36
6.4 Kt qu thc nghim ........................................................................................................ 38
7 Kt lun .................................................................................................................................. 40
Ti liu tham kho ........................................................................................................................ 42

H nano dn truyn thuc cho no

Trang 1

GVHD: PGS.TS L Th Hng Nhan

H NANO DN TRUYN THUC CHO NO

1 Gii thiu .
Glioma l loi ung th no thng gp nht, chim 29% ca tt c khi u thn
kinh trung ng v 80% cc khi u no c tnh. Thi gian sng trung bnh bnh nhn
glioblastoma (GBM) ch khong 14,6 thng sau khi iu tr phu thut ct b kt hp ha
tr v x tr ng thi [1]. Mt l do lin quan trc tip n vn kh chn on v ti
pht nhanh sau khi iu tr l s pht trin thm nhim ca gliomas gy kh khn cho
vic loi b hon ton m bnh hoc khu vc ung th xm nhp m khng nh hng
n s hot ng bnh thng ca no. Hn na, tc dng ph ca x tr v hu qu xu
ca cc phng php ha tr thng thng cng l nguyn nhn dn n s tht bi trong
iu tr [2].
Thuc truyn thng da trn c tnh hp th ca thuc i vi c th, do sinh
kh dng ph thuc nhiu yu t nh tan, pKa, khi lng phn t, s lng lin kt
hydrogen ca phn t, bn ha hc, tt c nhng yu t tc ng khng nh n
kh nng iu tr bnh. Thng thng bn cht thuc truyn thng l c khi lng phn
t nh, kh nng vt qua mng ngn ca c th v thm nhp vo cc bo quan cng
nh cc t bo. Do thuc ph hp cho vic cha bnh ung th. Tuy nhin hn ch v
phn b thuc dn n hiu ng ph khng mong mun khi phi s dng thuc vi liu
lng cao thuc c th p ng tt kh nng cha bnh. Bn cnh do khi lng
phn t nh, thuc d b o thi ra khi c th qua ng thn, c ngha l phi s dng
mt liu lng cao cha tr [3].
Trong nhiu nm, cc nh nghin cu c gng tm ra cc phng php mi
a cc tc nhn iu tr n khu vc khi u hiu qu v gim tch ly thuc khng cn
thit trong t bo no bnh thng v cc m ngoi vi. Nhiu nghin cu c tin hnh
c th m bo tt p ng thuc vi mt liu lng nh hn v hiu ng ph l nh
nht. Do , thuc cn phi ci thin sinh kh dng. Sinh kh dng nh hng bi nhiu
yu t, bao gm tnh cht vt l v tnh cht ha hc ca thuc, liu lng v nng s

H nano dn truyn thuc cho no

Trang 2

GVHD: PGS.TS L Th Hng Nhan

dng thuc cng nh chu k s dng. i vi cc khi u no, cch thng phn phi
thuc nh hng mc tiu theo c ch ch ng thu ht c s quan tm rng ri
trong nhiu thp k gn y. V khi u no c nhiu c im phc tp khc bit vi cc
khi u ngoi vi, nhiu yu t phi c xem xt phn phi thuc hiu qu, chng hn
nh cc ro cn trong ton b qu trnh, mi trng trong khi u v c im cc t bo
khi u [4]. Hin nay nhiu nh hng mc tiu khc nhau c khai thc t c
hiu qu iu tr cng vi vic s dng h thng phn phi kch thc nano. y , tc
gi s cung cp mt nh gi ngn gn v mt s mc tiu chin lc c th dng
phn phi thuc ti cc khi u no.
2 Cu to hng ro mu no
2.1 Lch s :
Hng ro mu no (Blood-Brain Barrier BBB) l mt mng sinh hc. Chng c
hai chc nng c bn: cung cp cc cht dinh dng (sinh hc) nui t bo no v
ngn cn cc cht c hi cho t bo no.
+ Chc nng dinh dng: cho php vn chuyn qua hng ro mu no nhng cht sinh
hc gip qu trnh chuyn ha ca t bo no.
+ Chc nng bo v: ngn cc cht c hi vn chuyn qua hng ro mu no.
Hng ro mu no bao gm cc t bo ni m (endothelial cell), t bo hnh sao
(astroglia), t bo ngoi mch (pericyte), cc i thc bo ngoi mch (perivascular
macrophage) v mng y (basal lamina), chng lin kt cht ch vi nhau. Vi c tnh
, chng hn ch cht ch s di chuyn cc cht t mu vo no, trong khi cc t bo
ni m mao mch cc b phn khc ca c th li khng c c c tnh .
Mt cu trc ca t bo hnh sao (astrocyte cell) nh ra, c gi l chn t bo hnh sao
(astrocytic feet) bao bc xung quanh cc t bo ni m ca hng ro mu no, to nn cc
c tnh sinh ha ca chng. Hng ro mu no c th phn bit vi cc hng ro tng
t nh chng l hng ro dch no ty- mu (blood-cerebrospinal fluid barrier: mt chc
nng ca cc t bo mng mch thuc m ri mch mc), v hng ro vng mc - mu

H nano dn truyn thuc cho no

Trang 3

GVHD: PGS.TS L Th Hng Nhan

(blood- retinal barrier). S pht hin hng ro mu no trong mt thi gian di v cng
ht sc tnh c. u tin, vo cui th k 19 nh khoa hc Paul Ehrlich trong khi tin
hnh th nghim c s ch n hng ro mu no. Paul Ehrlich l mt nh vi trng
hc, chuyn nghin cu cc cu trc c th thy c bng phng php nhum mu.
Khi ng tim cc cht mu vo c th, cc cht mu c th nhum tt c cc b phn ca
c th ngoi tr no b. Vo thi im , Ehrlich n gin cho rng no khng nhn cc
cht nhum mu. Sau , trong mt th nghim vo cui nm 1913, Edwin Goldmann
tim trc tip cht nhum mu vo dch no ty. ng ta pht hin rng, trong trng hp
ny no tr nn nhum mu, trong khi cc b phn cn li ca c th th khng bt mu.
iu ny chng t c s tn ti mt hng ro gia no v cc b phn cn li ca c th.
Vo thi im ngi ta cho rng, chnh h thng mch mu ng vai tr l hng ro
v hin nhin hng ro mu no vn cha c pht hin. Khi nim v hng ro mu
no c ngh bi Lina Stern vo nm 1921. Nhng mi cho n nm 1960, khi pht
hin ra knh hin vi in t, hng ro mu no mi c m t mt cch y . Ngi ta
cho rng, cc t bo hnh sao ch khng phi cc t bo biu m, ng vai tr c bn ca
hng ro mu no, bi v chn t bo hnh sao bao bc dy c quanh cc t bo biu m
ca hng ro mu no. Nm 1959, Hugh Davson pht hin ra hng ro mu- dch no ty.
2.2 Cu to
Hng ro mu no hnh thnh do cc t bo thn kinh hnh sao (astrocyte), chn
ca t bo ny bao bc ly mch mu dn truyn ln no. Bn cnh , mt ca
pericite cao (mt thnh phn ca mng mch mu) hnh thnh nn cc tight junctions lm
cho mng mch mu dn truyn ln no sp xp rt kht, hn ch s di chuyn ca cc
cht tan (hnh 1) . Theo nghin cu, mng mch mu ch cho cc cht dinh dng i qua
nh CO2, O2, glucose, cc cht a lipid, hn ch cc protein v cc polymer mch di
thm nhp qua mng mu. Bn cnh , cc cht c khi lng phn t di 50kDa cng
c kh nng cao qua mng mu no [5].

H nano dn truyn thuc cho no

Trang 4

GVHD: PGS.TS L Th Hng Nhan

Hnh 1: Cu to hng ro mu no
3 Kim sot thi gian gii phng thuc
Thng qua iu khin thi gian, cho php thuc t nng iu tr trong t bo.
Hnh 2 th hin nng thuc trong c th theo thi gian ca cc loi thuc thng
thng. Ban u nng thuc tng nhanh sau do qu trnh o thi qua thn v s
chuyn ha, trao i cht (thc bo,.enzyme..) nng thuc gim nhanh. Do cn
phi ch n vn o thi thuc [6].
Nng hiu qu l nng nh nht thuc t hiu qu iu tr. Trong khi ,
nng gii hn an ton l gi tr nng m khi vt qua n s gy c v tc dng
ph khng mong mun cho c th. Do vic thuc b o thi, ta phi s dng nhiu liu
m bo thuc lun t nng tr liu (hnh 2). Tuy nhin nu s dng khng ng
liu lng, gy ra s tch ly vt qu nng gii hn an ton s gy tc dng ph[6].

H nano dn truyn thuc cho no

Trang 5

GVHD: PGS.TS L Th Hng Nhan

Hnh 2 : Nng gii phng thuc theo thi gian ca liu n (tri) v liu a (phi)
3.1 Gii phng thuc dng Extended-Release formulation
S gii phng ca cc tc nhn iu tr trong cc cng thc ban u to ra mt
lng va t nng iu tr v sau tip tc gii phng thuc. Trong qu trnh
gii phng thuc, nng thuc gii phng khng nht thit l hng s nhng phi ko
di c thi gian duy tr nng hiu qu ( hnh 3) . C hai loi cng thc extendedrelease formulations. Th nht l dng polymeric matrix (hnh 3) c cha cc tc nhn
iu tr, s gii phng thuc c kim sot bng cch khuch tn qua cc khe h gia
cc chui polymer h thng mng matrix (s trng n v mt lin kt ngang quyt
nh n kh nng gii phng thuc). Nhng cng thc c p dng cho hydrogel v
nano particles. Th hai l dng polymeric capsules , s phn hy ca lp ph polymer
quyt nh vic gii phng ca thuc[7].

Hnh 3: Cc dng gii phng thuc v ng ng hc gii phng thuc ca

Extended-Release formulation

H nano dn truyn thuc cho no

Trang 6

GVHD: PGS.TS L Th Hng Nhan

3.2 Gii phng thuc dng Sustained-Release formulation
ng ng hc ca qu trnh gii phng thuc trong trng hp ny l bc
khng (nng gii phng ca thuc l hng s trong khong thi gian iu tr), hiu qu
thuc duy tr trong thi gian di. H thng dn truyn thuc ny bao gm mt lp
polymer bn thm cha thuc (hay cc tc nhn iu tr khc) v h polymer y . H
kim sot qu trnh gii phng thuc bng gradient y xuyn qua lp polymer bn thm.
Khi gp nc, polymer y ny (polymeric osmotic system) b hydrate ha, trng ln
dn n tng p sut v gii phng thuc ( hnh 4). Ngi ta iu chnh gradient thm ca
mng bn thm v mc hydrate ha ca polymeric osmotic system kim sot thi
gian gii phng thuc[8].

Hnh 4 : C ch gii phng thuc v ng ng hc gii phng thuc ca

Extended-Release formulation
3.3 Gii phng thuc dng Pulsatile-Release formulation
Mt dng kim sot thi gian l h dn truyn thuc p ng , thuc ch c gii
phng khi c th yu cu. Do , thuc s c gii phng gin on sau nhng
khong thi gian nht nh v lp i lp li lin tc n khi lng thuc ht. H dn
truyn loi ny gm 2 thnh phn trong mt cu trc ng vai tr sensor pht hin
thay i ca mi trng xung quanh v mt cu trc ng vai tr gii phng thuc (hnh
5).

H nano dn truyn thuc cho no

Trang 7

GVHD: PGS.TS L Th Hng Nhan

H dn truyn ny cho php nh thuc ng v tr, ng thi gian (c th cn) v
ng lng tc nhn iu tr . Do , chng ph hp cho vic iu tr nhng cn bnh
mn tnh [9].
Mt v d trong trng hp ny l h thng dn truyn insulin iu tr tiu ng
vi glucose oxidase ng vai tr sensor, khi lng ng trong mu tng cao, lng
glucose oxidase s chuyn glucose thnh glucomic acid, lm gim pH, khi lp
polymer s trng ln v insulin s c gii phng ra bi thuc c bao bc bi loi
polymer nhy pH (hnh 5). Khi lng thuc gii phng t c hiu qu iu tr lng
glucose trong mu gim lm pH cng gim xung (do qu trnh chuyn glucose thnh
glucomic acid bi glucose oxidase gim), lp polymer s tr li bnh thng , qu trnh
gii phng thuc s ngng li[10].

Hnh 5: Mt loi polymer nhy pH v ng ng hc gii phng thuc ca PulsatileRelease formulation

4 Kim sot v tr gii phng thuc theo c ch ch ng
Cu to ca u thn kinh m rt phc tp vi nhiu loi ro cn sinh l khc nhau
ngn chn thuc n v tr khi u. C hai ro cn chnh cho vic iu tr khi u no: hng
ro mu no (BBB) v hiu ng EPR tng i yu [11].

H nano dn truyn thuc cho no

Trang 8

GVHD: PGS.TS L Th Hng Nhan

Hng ro mu no (BBB) l mt h thng c cu to c bit vi cc t bo ni
m mao mch c bao ph mt phn bi pericytes v mng nn v c bao quanh gn
nh hon ton bi chn ca t bo hnh sao, n ngn chn khong 98% ca cc phn t
nh v gn nh 100% cc phn t ln bao gm cc protein ti t hp , thuc v gen c
vn chuyn vo no n v tr khi u. Hng ro mu no kim sot nghim ngt s vn
chuyn thuc vo trong no bng hng ro vt l (tight junctions) , s chuyn ha cht
(metabolic enzyme) v hng ro min dch (immunological barrier)[12].
gii quyt thch thc ny, nhiu loi chin lc nh hng mc tiu theo c
ch ch ng c p dng pht trin hiu qu h thng phn phi thuc n no
[13]. Cc h thng nhm mc tiu theo c ch ch ng ch yu c chia thnh: hp th
trung gian transcytosis (absorptive mediated transcytosis), vn chuyn trung gian
transcytosis ( transporter-mediated transcytosis) v th th trung gian endocytosis
(receptor-mediated endocytosis) (hnh 6).

Hnh 6: Cc con ng dn truyn thuc vo no

H nano dn truyn thuc cho no

Trang 9

GVHD: PGS.TS L Th Hng Nhan

4.1. Con ng dn truyn absorptive-mediated transcytosis
Hp th trung gian transcytosis l phng php phn phi thuc qua hng ro mu
no bng protein cation hoc cell-penetrating peptides (CPPs). Qa trnh phn phi thuc
c thc hin bi tng tc tnh in gia nhng v tr tch in dng cc gc protein
v b mt mng t bo trn cc t bo ni m no tch in m (cu to cc acid amine
tch in m) (hnh 7) [14].

Hnh 7: Cu trc cc acid amine tch in

S dn truyn thuc qua hng ro mu no (BBB) bng cationic proteins
H

dn

truyn

cationic

bovine

serum

albumin

(CBSA)

kt

hp

poly(ethyleneglycol)-poly(lactide) (PEG-PLA) nanoparticle (CBSA-NP) c tng

hp bi Lu v cc ng s dn truyn thuc vo no theo c ch ch ng. CBSA
ng vai tr nh mt "brain targetor" c hiu, c lin kt ha tr vi nhm chc
maleimide cui ca poly (ethyleneglycol) (PEG) bao quanh ht nano. PEG c chn
lm phn khc to v bi nhng c im l ha ca n bao gm tan trong nc tt ,
s linh ng ca chui ln cng nh c tnh thp , tng thi gian tun hon , ci thin s
tch ly thuc , nh tng cng hiu ng EPR. Mt khc, copolymers c kh nng to
ht nano trong vng kch thc mesoscopic (vi chc nm) vi khong phn b kch thc
hp [15].
Cc ht nano (NPs) to thnh t hn hp ca MPEG-PLA v maleimide-PEGPLA (copolymers) c tng hp bng phng php nh ha / bay hi dung mi. CBSA

H nano dn truyn thuc cho no

Trang 10

GVHD: PGS.TS L Th Hng Nhan

c iu ch t BSA qua qu trnh cation ha vi ethylenediamine trong EDAC. CBSA
sau c phn ng vi 2-iminothiolan (Trauts reagent). Cc thiolated CBSA
(CBSA vi nhm SH va thu c) c trn vi cc ht nano theo t l CBSA
thiolated: maleimide l 1: 1 (gn CBSA thiolated vo ht nano ph PEG thng qua
maleimide). Cc nhm amino trong cu trc CBSA-NP c im ng in cao nn trong
mi trng sinh l c th chng tn ti dng cation (hnh 8) [15].

Hnh 8: Cu trc PEG-PLA- CBSA-NP

Kch thc ht l mt tnh cht quan trng ca h nano c nh hng trc tip n
kh nng dn truyn thuc, s phn b kch thc ng knh ht thng c kim sot
di 200 nm. Trong nghin cu ny, kch thc h nano tng hp c c ng knh
di 100 nm to iu kin thun li cho vic vn chuyn thuc qua hng ro mu no
Kch thc ht nano khng thay i ng k khi kt hp CBSA , thm ch c sau khi np
thuc vo ht nano (bng 1).

H nano dn truyn thuc cho no

Trang 11

GVHD: PGS.TS L Th Hng Nhan

Bng 1 : S phn b kch thc ca NPs v CBSA-NPs (c np hay khng np 6coumarin)

Kh nng thm (permeability) ca CBSA-NP trong mi trng ni bo c kho
st v so snh vi bovine serum albumin (BSA) gn vi ht nanoparticles (BSA-NP).
CBSA-NP khng nh hng n cu trc ca knh ( tight junctions ) trong hng ro mu
no. CBSA-NP c c tnh thp i vi t bo ni m mao mch no ( BCECs). Kh
nng thm ca CBSA-NP cao hn khong 7,76 ln so vi ca BSA-NP , trong khi qu
trnh transcytosis b c ch vi lng d CBSA t do (bng 2). C th kt lun rng
CBSA-NP c kh nng vt qua BBB tt vi c tnh thp, to iu kin thun li
cung cp cc tc nhn tr liu ti thn kinh trung ng [15].

Bng 2: Kh nng thm ca BSA-NP v CBSA-NP qua hng ro mu no.

H nano dn truyn thuc cho no

Trang 12

GVHD: PGS.TS L Th Hng Nhan

Kh nng gii phng thuc ca cc ht CBSA-NP cha 6- coumarin c xc nh
bi th nghim th leaching ca thuc nhum trong mui sinh l PBS 0,1M iu kin
khc nhau (pH 4.0 v 7.4). Kt qu thc nghim cho thy khong 0,48% ca thuc
nhum c gii phng t CBSA-NP pH 4,0 so vi khong 0,72% pH 7.4 trong
72 gi khi ngm trong dung dch mui sinh l (hnh 9). iu ny cho thy lng thuc
gii phng thuc xy ra tt trong mi trng pH ni bo v thi gian gii phng c ko
di ng k [15].

Hnh 9: ng ng hc gii phng 6- coumarin t CBSA-NP pH=4 v pH=7.4

Tc gi dng phng php pht x hunh quang xc nh kh nng cc ht
nano vt qua c hng ro mu no, s phn b ca 6- coumarin trong vng i no
ca chut c theo di lin tc 4 h sau khi tim . BSA-NP v NPs khng th vt qua
hng ro mu no (mi tn trng). Ngc li, CBSA-NP c kh nng vt qua hng ro
mu no v tp trung xung quanh cc ht nhn ca t bo thn kinh ( cc mi tn trng)
(hnh 10). Cc ht nano khng thm nhp vo nhn t bo ca t bo thn kinh c th v
kch thc ca cc ht nano rt ln , trong khi BSA-NP khng th qua hng ro mu no
do chng khng tch in [15].

H nano dn truyn thuc cho no

Trang 13

GVHD: PGS.TS L Th Hng Nhan

Hnh 10: Kho st kh nng dn truyn vo no ca BSA-NP (hnh trn) v

CBSA-NP (hnh di) bng pht x hunh quang
S dn truyn thuc qua hng ro mu no (BBB) bng cell penetratingpeptides (CPPs)
H thng dn truyn thuc da vo AMT-type cell-penetrating peptide (CPP) cho
thy kh nng phn phi thuc qua hng ro mu no rt tt . CPPs c s dng vt
qua nhng ro cn lipophilic ca mng t bo v mang theo mt lng ln cc tc nhn
iu tr, trong c peptide / protein, DNA / oligonucleotide, khng th, v cc h thuc
nano nh liposomes, micelles,...
CPPs khng ng nht v kch thc v trnh t chui tch in dng. Mt s c
tnh nng ph bin ca CPPs nh mt chui amphipathic v kh nng tng tc vi mng
lipid.CPPs thng c ngun gc t protein t nhin bao gm transcription-activating
factor Tat , penetratin v Syn-Bvectors, trong Tat c s dng thng xuyn nht
[16].

H nano dn truyn thuc cho no

Trang 14

GVHD: PGS.TS L Th Hng Nhan

Chui peptide TAT l mt loi CPP cha nhiu nhm arginine. TAT t bn ngoi
c th nhp c cc t bo khng b nhim v b nhim . Tuy nhin cc c ch phn t
chnh xc m theo TAT i vo cc t bo vn cn cha r rng. TAT lin kt vi b
mt t bo heparin sulphate v cc lin kt gia ton b chiu di protein TAT vi c
proteoglycans heparin sulphate v h low - density lipoprotein receptor c chng
minh. Chui TAT c bn gm mt cm cation gm 6 arginine v lysine 2 ng vai tr
quan trng trong s vn chuyn ca cc protein qua mng sinh hc; qu trnh chuyn v
c h tr bi s kt dnh mnh m ca cc t bo vi cu trc khung ny, s rng buc
t bo c lp vi t bo th th v nhit . Cc nhm u guanidinium ca arginine cn
thit cho s hp thu peptide v tch in dng mnh hn cc nhm cation khc (do s
khc nhau ca im ng in), chng hn nh lysine, histidine hoc ornithine (hnh 11).

Hnh 11: Cu trc arginine

TAT peptide (TAT-PEG-b-Col) c tng hp bi Liu v cc ng s dn
truyn khng sinh qua hng ro mu no. Cholesteryl cloroformat ha tan trong DCM
c thm t t ,ng thi khuy mnh vo mt dung dch cha TEA v NH2-PEGCOOH. Hn hp ny phn ng trong 2 ngy. Sn phm th c tinh ch bng

H nano dn truyn thuc cho no

Trang 15

GVHD: PGS.TS L Th Hng Nhan

phng php thm tch loi b DCM trong 6 ngy s dng mng vi khi lng phn t
2000 DA. TAT peptide vi chui NH2-YGRKKRRQRRR c lin kt vi PEG thng
qua nhm carboxylic acid (hnh 12) [17].

Hnh 12: Cu trc TAT-PEG-b-Col

Cc ht nano c tng hp c mt li k nc ca cholesterol cha thuc v mt
lp v a nc ca PEG c cha cc phn t TAT tng cng s hp thu thuc vo
no. Nano polyme c cu trc li / v l h dn truyn y tim nng phn phi thuc
. Copolymer c kh nng to ht vi kch thc nh (<200 nm) vi khong phn b kch
thc hp v c th ha tan thuc k nc thng qua s tng tc k nc hoc lin kt
hydro trong li bao quanh bi lp v a nc bn ngoi. Cu trc ny c hiu qu bo v
cc cht c hot tnh sinh hc , ngn chn s phn hy thuc ko di tun hon trong c
th bng cch trnh qu trnh thc bo ca h thng li ni m (RES) [17].
Hnh dng v kch thc cc ht nano blank v nano cha ciprofloxacin c xc
nh bng cch s dng DLS v SEM. Cc ht nano l c dng hnh cu vi kch thc
trung bnh thp hn 200 nm. Kch thc ht nano khng thay i ng k khi kt hp
TAT , thm ch c khi np thuc vo ht nano ( tuy nhin kch thc ny vn cn ln
c th vt qua hng ro mu no) (hnh 13). Kch thc ca cc ht nano trong hnh

H nano dn truyn thuc cho no

Trang 16

GVHD: PGS.TS L Th Hng Nhan

SEM nh hn mt cht so vi cc kt qu thu c t phng php nh sng tn x
nguyn nhn l do s co li ca lp v cc ht nano trong qu trnh sy chn khng qu
trnh chun b mu SEM [17].

Hnh 13: Hnh SEM ca TAT-PEG-b-Col

Kt qu thc nghim cho thy profile gii phng thuc t cc ht nano trong dung
dch mui sinh l PBS (pH 7.4) 370C. Qa trnh gii phng thuc c duy tr trong
mt khong thi gian 5 h, iu ny chng t phn t thuc c c bao bc tt vo cc
ht nano (hnh 14) [17].

Hnh 14: ng ng hc gii phng ciprofloxacin t TAT-PEG-b-Col

H nano dn truyn thuc cho no

Trang 17

GVHD: PGS.TS L Th Hng Nhan

S phn b ca ciprofloxacin trong vng i no ca chut c theo di lin
tc bng phng php pht x hunh quang. S hp thu cc ht nano lin kt vi TAT
bi cc t bo ni m no ln hn so vi cc ht nano khng gn TAT. Quan trng nht,
cc ht nano TAT-PEG-b-Chol c th vt qua hng ro mu no (BBB) v tp trung
xung quanh nhn t bo ca t bo thn kinh , trong khi PEG-b-Chol khng th vt
qua hng ro ny. Nhng nghin cu ny cho thy kh nng dn truyn ca cc h Tatmodified nanoparticle vo no vi mc ch chn on v iu tr cc bnh lin quan n
no (hnh 15) [17].

Hnh 15: Kho st kh nng dn truyn vo no ca PEG-b-Chol (hnh trn) v

TAT-PEG-b-Chol (hnh di) bng pht x hunh quang
4.2 Con ng dn truyn transporter-mediated transcytosis
Bi v c rt nhiu loi h thng vn chuyn trn ni m no cung cp cho no
cc cht dinh dng cn thit v cc cht ni sinh, vn chuyn trung gian transcytosis

H nano dn truyn thuc cho no

Trang 18

GVHD: PGS.TS L Th Hng Nhan

(transporter mediated transcytosis) s dng cc h thng vn chuyn ny nh mt
phng php nh hng mc tiu vo cc m no y tim nng . Transporter mediated
transcytosis l phng php dn truyn da trn cht nn c tnh chn lc, ch nhng tc
nhn iu tr c tnh cht gn ging cc cht ni sinh hay cc cht dinh dng cn thit
mi c kh nng i qua v vn chuyn vo trong m no [18].
Cc GLUT1 phn b mt cao trn mng ca t bo ni m no BCECs. Hn
na, tiu th glucose ca no chim khong 30% tng lng tiu th glucose ca c th.
Mc hp thu glucose cao ca no cho thy -D glucose transporter GLUT1 l mt h
thng vn chuyn tt cho glucose nh hng mc tiu dn truyn thuc hiu qu v
c chn lc vo no [18].
Vi phng php ny, Qin v cc ng s tng hp mt loi dn xut glycosyl
mi ca cholesterol lm vt liu to ra mt loi liposome mi, khc phc vn phn
phi khng hiu qu vo t bo no ca cc cng thc liposome thng thng ny bng
cch nh hng mc tiu vo cc h thng cht vn chuyn glucose trn hng ro mu
no. Trong cu trc ny nhng nhm steroid a du nm chn vo mng liposomal v cc
nhm glucose a nc nm bn ngoi mng s c nhn din bi GLUT1 v vn
chuyn qua hng ro mu no. Cholesterol lin kt vi ng glucose thng qua glycols
ethylene v tr C-6 ca glucose nhm to i lc cao vi GLUT1 (hnh 16). Ethylene
glycol c kh nng lin kt vi cc th th sinh hc v cc ligand, v lm gim s lin kt
khng c hiu ca protein v phn t c hot tnh sinh hc khc. Mt khc, PEG thng
c s dng trong cc h thng dn truyn thuc bi nhng c im tnh cht l ha
ca n bao gm tan trong nc tt , s linh ng ca chui ln cng nh c tnh
thp , tng thi gian tun hon , ci thin s tch ly thuc , nh tng cng hiu ng
EPR [19] .

H nano dn truyn thuc cho no

Trang 19

GVHD: PGS.TS L Th Hng Nhan

Hnh 16: Quy trnh tng hp v cu trc GLP-4

Hnh dng v kch thc cc ht liposome blank v liposome cha Coumarin-6
c xc nh bng cch s dng DLS v SEM. Cc ht nano l c dng hnh cu vi
kch thc trung bnh thp hn 150 nm. Kch thc ht liposome khng thay i ng k
khi kt hp nhm glucose , thm ch c khi np thuc vo ht liposome (hnh 17). iu
ny rt c ngha trong vic thit k cc h nano trong dn truyn thuc. Kch thc ht
ng mt vai tr quan trng quyt nh kh nng chng b o thi bi l lch ca ngi
v chut. Ht phi nh trnh qu trnh lc ti cc khe t bo interendothelial (IES)
trong thnh tnh mch . Tuy nhin, kch thc cc khe ny t khi vt qu 200-500 nm .
Do , kch thc ca ht khng c vt qu 200 nm . Nu ln hn, ht phi bin
dng vt qua IES hay khng th vt qua hng ro ny .V vy, kch thc ht l
mt tnh cht quan trng ca h nano c nh hng trc tip n kh nng dn truyn
thuc, s phn b kch thc ng knh ht thng c kim sot di 200 nm khng
ch cho ht nano m c nhng immunoliposomes. Trong nghin cu ny, kch thc h

H nano dn truyn thuc cho no

Trang 20

GVHD: PGS.TS L Th Hng Nhan

liposome tng hp c c ng knh di 150 nm to iu kin thun li cho vic vn
chuyn thuc qua hng ro mu no [19].

Hnh 17: Ph phn b kch thc ht ca LIP (tri) v GLP-4 (phi)

nh gi c th kh nng vn chuyn qua hng ro mu no ca liposome lin
kt vi glucose , s phn phi trong cc m no ca LIP (liposome thng thng), CLP
(liposome bin tnh), v GLP-4 (liposome gn nhm glucose ) c kho st. S phn
b cc m ca coumarin-6 c o sau 5, 15, 30 pht, 1 gi, 2 gi, 4 h, 8 h, 12 h v 24 h
sau khi tim. Cc thng s dc ng hc (RE v CE) ca coumarin-6 trong no c
ghi nhn trong bng . Trong no, ch s AUC0-t ca coumarin-6 trong GLP-4 cao hn
2,85 ln so vi LIP, v cao hn 3,33 ln so vi CLP. Cc Cmax ca GLP-4 cao hn so
vi LIP 1,43 ln v 3,10 cao hn ln so vi CLP (bng 3) [19].

H nano dn truyn thuc cho no

Trang 21

GVHD: PGS.TS L Th Hng Nhan

Bng 3: Cc thng s dc ng hc ( RE v CE) ca coumarin-6 trong no

Kt qu thc nghim cho thy profile gii phng thuc t cc ht liposome LIP,
CLP, v GLP-4 trong no. Qa trnh gii phng thuc ca cc loi liposome c duy tr
trong mt khong thi gian di12 h, chng t phn t thuc c c bao bc tt vo
cc ht liposome. Trong , kh nng gii phng thuc ca GLP-4 l tt nht ( vi nng
t hn 200ng/g sau 12 h) (hnh 18) .iu ny cho thy lng thuc gii phng thuc
ca GLP-4 xy ra rt tt trong mi trng t bo ni m no v thi gian gii phng
c ko di ng k [19].

Hnh 18: Kho st nng thuc ca LIP, CLP, v GLP-4 trong no

H nano dn truyn thuc cho no

Trang 22

GVHD: PGS.TS L Th Hng Nhan

Ngoi ra, Quin v cc ng s cng kho st kh nng vn chuyn qua hng ro
mu no bng cch dng pht x hunh quang kim tra lng coumarin-6 trong BCECs
(hnh 19). Cc nhn t bo c nhum mu xanh vi DAPI . S pht hunh quang
mu c tm thy trong nhn t bo v t bo cht. Trong s tt c ba loi liposome
kho st, s dn truyn thuc vo BCECs no ca liposome gn nhm glucose l tt nht
(Hnh.). S pht hunh quang (mu ) mnh nht c quan st thy trong t bo
BCECs iu tr bng GLP-4 . Cng pht hunh quang (mu ) mnh nht tm thy
trong nhn t bo v t bo cht. Cng pht hunh t bo BCECs iu tr bng
gim ng k [19].

Hnh 19: Kho st kh nng dn truyn vo no LIP, CLP, v GLP-4 (theo th t t tri
sang phi) bng pht x hunh quang
4.3 Con ng dn truyn receptor-mediated transcytosis
Ni chung, tnh c hiu v chn lc ca con ng hp th qua trung gian
transcytosis ( absorptive-mediated transcytosis) nh hng mc tiu BBB khng cao
v cc cationic proteins hoc CPPs c th tng tc v lin kt vi bt k thnh phn no
c mng t bo tch in m trong c th. Hn na, c tnh v s chng min dch ca
cation protein hoc CPP lm hn ch ng dng trong dn truyn thuc. Trong khi ,con
ng vn chuyn qua trung gian transcytosis ( transporter-mediated transcytosis) da
trn cht nn c tnh chn lc, ch nhng tc nhn thuc c tnh cht gn ging cc cht

H nano dn truyn thuc cho no

Trang 23

GVHD: PGS.TS L Th Hng Nhan

ni sinh hay cc cht dinh dng cn thit mi c kh nng i qua v vn chuyn vo
trong m no [20].
Vn chuyn qua th th trung gian transcytosisis c coi l mt trong nhng cc
chin lc dn truyn thuc c nh hng cho no hon thin nht vi tnh c hiu cao,
chn lc tt v i lc vi mc tiu ln, mc d ligand c th c nh hng n trng thi
cn bng ni mi trong c th v ligand t nhin c th cnh tranh vi thuc gim
hiu qu nh hng v phn phi ca h [20].
C nhiu loi th th trn nhng mao mch ni m ca no, chng hn nh th th
transferrin (TFR), low-density lipoprotein (LDLR), th th insulin v th th nicotinic
acetylcholine, cc loi ligand nhm mc tiu, bao gm c ligand c tnh ni sinh v
ligand thc khun hay cc khung cu trc nh hng. Cc loi th th ny c s
dng dn truyn thuc qua hng ro mu no. Mt trong nhng h thng th th
transcytosis c s dng rng ri nht dn truyn thuc vo no l th th transferrin
(TFR) c phn b dy c trn cc t bo ni m ca hng ro mu no [20].
St l mt c cht quan trng ca nhiu loi enzyme khc nhau cn thit cho nhiu
qu trnh trao i cht, bao gm tng hp DNA, chuyn in t, vn chuyn v lu tr
oxy . T bo hp thu ion st t mi trng ngoi bo qua th th trung gian endocytosis ,
nhng st khng tng tc trc tip vi nhng th th . Thay vo , st lin kt vi
nhng protein transferrin, sau nhng protein ny lin kt vi cc th th transferrin v
vn chuyn st vo mi trng ni bo. y chnh l c s quan trng thit k h dn
truyn thuc theo con ng hp th qua trung gian transcytosis ( absorptive-mediated
transcytosis) qua th th transferrin (TFR) [20].
Cu trc protein c bn ca transferrin c to thnh t khong 700 n v axit
amin (80 kDa). Trong transferrin c s kt hp ca xon alpha v lp beta to thnh 2
nhnh khc nhau: nhnh N v nhnh C. Hai nhnh ny c gi li vi nhau bi mt
peptide ngn v to ra v tr k nc . Cc loi axit amin lin kt vi ion St ging nhau
c hai nhnh , bao gm hai phn t tyrosine, mt phn t acid aspartic v mt phn t

H nano dn truyn thuc cho no

Trang 24

GVHD: PGS.TS L Th Hng Nhan

histotine. S lin kt ca st vo hai nhnh ca transferrin cn mt anion, thng l
ion cacbonat (CO32-). in tch 3+ ca ion st c n nh bi hai phn t tyrosine v
mt phn t axit aspartic. Cc in tch trn anion c cn bng ca in tch dng
lin k trn protein (trong transferrins, phn t tch in dng xut pht t nhng chui
arginine v N-termins ca chui xon alpha) (hnh 20).

Hnh 20: Cu trc transferrins

Zhang v cc ng s tng hp thnh cng h polyphosphoester hybrid micells
gn transferrin dn truyn paclitaxel qua hang ro mu no. Maleimide-PEG-OH c
tng hp bng phn ng gia NH2-PEG-OH v 3-(maleimido) propionic acid Nhydroxysuccinimide ester (MPA-SE). Sau , Maleimide-PEG-PCL c tng hp bng
phn ng trng hp m vng ca -caprolacton v maleimide-PEG-OH vi Sn (Oct)2 l
cht xc tc. tng hp hybrid micells gn Tf dn truyn paclitaxel (TPM), Tf u
tin c gn nhm S-H trc khi gn vo lin kt i ca nhm maleimide trong
Maleimide-PEG-PCL (hnh 21). Cc polyphosphoester hybrid micelle hnh thnh cu
trc li-v trong dung dch nc vi hiu qu np thuc cao (89,9 3,4%). TPM thu
c c kch thc trung bnh 87,85 2,32 nm vi th l -12,33 1,46 mV [21].

H nano dn truyn thuc cho no

Trang 25

GVHD: PGS.TS L Th Hng Nhan

Hnh 21: Cu trc polyphosphoester hybrid micells gn transferrin (TPM)

ng hc ca qu trnh gii phng paclitaxel (PTX) t PM v TPM c o trong
dung dch sodium salicylate 1 M 37 C, trong tan ca PTX tng ln n 28,1 mg /
mL m khng ph v cu trc ca PEG-b-poly ( phenylalanine) micelle . Kt qu thc
nghim cho thy PTX c gii phng n nh t TPM. Hn 90% PTX c gii phng
t cc micelle trong vng 72 h, trong khi , vic gii phng PTX t Taxol (h
polyphosphoester hybrid micells khng gn transferrin ) dn truyn paclitaxel thu
c t hn 80% trong cng khong thi gian.Ngoi ra, tc gi cng dng phng php
pht x hunh quang xc nh kh nng cc ht nano vt qua c hng ro mu
no, s phn b ca PTX trong vng i no ca chut c theo di lin tc trong 4
ngy sau khi tim. PTX trong Taxol khng th vt qua hng ro mu no. Ngc li,
PTX trong TPM c kh nng vt qua hng ro mu v tp trung nng cao trong no
thm ch sau 4 ngy (hnh 22) [21].

H nano dn truyn thuc cho no

Trang 26

GVHD: PGS.TS L Th Hng Nhan

Hnh 22: ng ng hc gii phng paclitaxel ca Taxol ,TPM v kho st kh nng

dn truyn vo no ca Taxol (con chut bn tri trong mi hnh) , TPM (con chut
bn phi trong mi hnh) qua pht x hunh quang
TPM cho thy kh nng tch ly thuc trong no tt hn Taxol, thi gian sng
trung bnh ca Taxol v TPM ln lt l 33,6 v 39,5 ngy. Thi gian sng st ca
chut mang khi u iu tr bng Taxol khng di (35 ngy). Ngc li, TPM ko di
ng k thi gian sng st

ca nhng con chut mang khi u ln ti 45 ngy

(p<0.01).Cc kt qu trn cho thy TPM c th l h dn truyn nano tim nng cho no
(hnh 23) [21].

H nano dn truyn thuc cho no

Trang 27

GVHD: PGS.TS L Th Hng Nhan

Hnh 23: Kho st thi gian sng ca chut mang khi u no trong th nghim khi iu
tr bng Taxol v TPM
4.4 H dn tryn thuc nhy cm mi trng
4.4.1 H dn tryn thuc nhy cm vi nhit
Trong nghin cu dn truyn thuc, polymer nhy nhit c l l loi c
nghin cu nhiu nht trong nhng h polymer nhy cm vi mi trng. Polymer nhy
nhit c chia l 2 loi: Polymer thun nhit (positively thermosensitive) v Polymer
nghch nhit (negatively thermosensitive). Cc loi polymer thun nhit tng tnh i nc
khi nhit tng lm trng n cu trc, trong khi cc polymer nghch nhit gim tnh
i nc khi nhit tng khin chng co li [22].
t c tnh cht nghch nhit ny , cc polymer phi c t l ph hp gia
cc chui i nc v k nc trong cu trc ca n, v d nh polymer trong nc nhit
phng to lin kt hydro ( ti cc tm a nc), nhng khi nhit tng ln cc lin
kt hydro ny b ph v , tng tc k nc lc ny chim u th v cu trc polymer co
li. Nu t l nhm k nc trong cu trc polymer tng ln th nhit im chuyn pha
t trng thi co sang trng thi trng n tng ln. Nh vy, bng cch kim sot t l

H nano dn truyn thuc cho no

Trang 28

GVHD: PGS.TS L Th Hng Nhan

gia cc chui i nc v chui k nc trong cu trc polymer , ngi ta c th iu
khin c s gii phng thuc cc iu kin khc nhau (hnh 24) [23].

Hnh 24: Mt s loi polymer thun nhit v nghch nhit

4.4.2 H dn tryn thuc nhy cm vi pH
Cc loi polymer ny c c trng bi s hin din ca nhm c tnh acid
,chng hn nh acid cacboxylic hoc acid sulfonic, hoc cc nhm c tnh base, chng
hn nh cc amin, trong cu trc ca chng. Nhng nhm ny c th nhn hoc cho
proton ph thuc vo s thay i pH ca mi trng, dn n s ion ha trong cu trc
ca polymer. Nhng polyme c nhiu nhm ion c gi l polyelectrolyte [24].
i vi cc polyme c cha nhm mang tnh acid , khi pH ca tng, mc ion
ha polymer tng ln. Ngc li, i vi polymer c cha nhm amino trong cu trc ,
khi pH ca tng ln, mc ion ha polymer gim . S thay i mc ion ha gy ra
s thay i trng n ca polymer do lc y tnh in gia cc tm ion trong cu
trc v thuc c gii phng. Trong loi polymer ny, vic kim sot qu trnh gii
phng c thc hin bng vic iu chnh tnh cht v s lng cc tm ion (hnh 25)
[25].

H nano dn truyn thuc cho no

Trang 29

GVHD: PGS.TS L Th Hng Nhan

Poly(axit acrylic) (PAA) b ion ha pH cao, trong khi poly(N,Ndiethylaminoethyl methacrylate) (PDEAEM) b ion ha pH thp. Nh vy, h dn
truyn polymer PAA gii phng thuc pH cao ( mi trng base) cn h dn truyn
PDEAEM gii phng thuc pH thp ( mi trng acid) [26].

Hnh 25: C ch ion ha ca cc loi polymer nhy cm pH

4.4.2 H dn tryn thuc nhy cm vi nh sng
Cc loi polymer ny c c trng bi s hin din ca nhm c kh nng phn
tch thnh cc cp ion khi tng tc vi nh sng UV (polymer nhy cm nh sng
UV)hay trng n khi b kch thch bi nh sng nhn thy (polymer nhy cm nh sng
nhn thy). H polymer nhy cm nh sng c nhng u im so vi cc loi polymer
nhy nhit (tnh nhy b nh hng bi s khuch tn nhit) v polymer nhy pH (tnh
nhy b nh hng bi s khuch tn ion hydro). Cc loi polymer nhy cm vi nh
sng UV c tnh hp bng cch a cc dn xut ca leuco (bis(4-dimethylamino)phenylmethyl leucocyanide) vo trong cu trc polymer .Dn xut ca
Triphenylmethane leuco bnh thng trung tnh nhng khi tng tc vi nh sng UV
chng phn tch thnh ion triphenylmethyl cations, polymer lc ny trng n do lc y
tnh in gia cc tm ion v thuc c gii phng (hnh 26) [27].

H nano dn truyn thuc cho no

Trang 30

GVHD: PGS.TS L Th Hng Nhan

Hnh 26: C ch ion ha ca dn xut bis(4-di- methylamino)phenylmethyl leucocyanide

Thc t, cc loi polymer nhy cm vi nh sng nhn thy c u tin pht trin
hn do ngun nh sng nhn thy an ton hn, ph bin, khng tn km. Loi polymer
ny c tng hp bng cch a nhm light-sensitive chromophore ( v d nh
trisodium salt of chlorophyllin) vo cu trc polymer. Khi tng tc vi nh sng nhn
thy chromophore s hp thu nng lng v to ra nhit cc b lm cu trc polymer
trng n v gii phng thuc [28].
5 Kim sot v tr gii phng thuc theo c ch th ng (hiu ng EPR)
Trong nhng nm gn y, cc h dn truyn polyme thng minh ca thuc chng
ung th c nghin cu rng ri . Cc loi polyme ny c th tch ly th ng trong
t bo ung th do s khc bit v c im sinh ha v sinh l gia m lnh tnh v c
tnh. Ni chung, cc phn t nh c th khuch tn qua mng t bo ni m ca c m
khe mnh v m c tnh. Tuy nhin, cc i phn t ch c th i vo m c tnh v
thnh t bo ni m ca cc m ny sinh trng mnh, chen ln to thnh cc l hng c
kch thc ln cho php cc i phn t vt qua d dng. Cc tnh nng khc ca m
c tnh nh dn lu bch huyt km v tng sinh mch mu (qu trnh angiogenic), u
dn n vic tng cng lu gi cc i phn t. iu ny to ra s tch t th ng ca
cc i phn t trong m khi u, c gi l hiu ng EPR (enhanced permeability and
retention effect) (hnh 27) [29].

H nano dn truyn thuc cho no

Trang 31

GVHD: PGS.TS L Th Hng Nhan

Hnh 27: Hiu ng EPR (enhanced permeability and retention effect)

Khi cc i phn t tip cn m c tnh, chng c thc hin qu trnh ni bo
endocytosed (receptor-mediated endocytosis) hoc phagocytosed (ni bo khng trung
gian receptor) bi cc m t bo c tnh to thnh cc ti ni bo. Do pH trong cc ti
thp (6.5- 5.0) so vi cc mi trng ngoi bo (7,2-7,4), cc mi lin kt khng bn vi
mi trng acid c thy phn v gii phng thuc. Cui cng, ti ni bo c hp th
bi mt lysosome dn n vic to thnh lysosome th cp c cha cc enzyme thy
phn v tiu ha hot ng trong mi trng axit (pH lysosome = 4). Mt s nghin cu
ch ra rng trong m c tnh , mt s loi enzym c trng c nng vt qu mc bnh
thng. Nhiu h dn truyn tn dng c im ny kim sot qu trnh gii phng
thuc bng cch s dng cc lin kt khng bn vi cc enzyme c trng ny . Hn na,
pH ca m khi u thp hn hn so vi cc m khe mnh (s tch t ca acid folic) v
do c th kim sot qu trnh phn phi thuc da vo cc loi lin kt khng bn
trong mi trng acid [30].
khai thc hiu qu hiu ng EPR, cc nghin cu tp trung pht trin cc h
dn truyn nano to bi block copolymer do nhng tnh cht c o ca chng. Cc h
dn truyn copolymer to nn cu trc li-v , c kh nng mang nhiu loi tc nhn iu

H nano dn truyn thuc cho no

Trang 32

GVHD: PGS.TS L Th Hng Nhan

tr bao gm c genes v proteins vi hiu qu np thuc cao. ng lc ca qa trnh
micelle ha cc copolymer c th l tng tc k nc ( do s khc bit ln v tan ca
phn khc to li v phn khc to v) , tng tc tnh in (polyion complex micelles)
hay s to phc kim loi [31].
So vi micelles hot ng b mt, cc micelles polymer n nh hn vi nng
CMC thp hn v mc phn tch chm hn. iu ny cho php duy tr qu trnh phn
phi thuc trong mt thi gian di hn v t c s tch ly cao hn ca thuc ti v tr
mc tiu. Hn na, cc coplymer c th to ht trong vng kch thc mesoscopic (vi
chc nanomet) vi s phn b kch thc trong mt khong hp. Phm vi kch thc hp
ca cc ht nano to ra t cc loi copolymer ny tng t nh kch thc ca virus v
lipoprotein v l mt yu t rt quan trng trong dn truyn thuc (c bit l vo no),
nng cao tc dng hiu ng EPR [31].
PEG thng c s dng lm phn khc to v ca copolymer trong cc h
thng dn truyn thuc bi nhng c im tnh cht l ha ca n bao gm tan
trong nc tt , s linh ng ca chui ln cng nh c tnh thp , tng thi gian tun
hon , ci thin s tch ly thuc ti cc v tr mc tiu, nh tng cng hiu ng EPR
.Mt khc , PEG c th d dng bin tnh v lin kt c vi nhiu loi nhm chc ,
protein, peptide dn truyn thuc theo c ch ch ng (hnh 28) [31].

H nano dn truyn thuc cho no

Trang 33

GVHD: PGS.TS L Th Hng Nhan

Hnh 28:Cu trc cc loi copolymer ph bin (tri) v cch thc tng hp cc loi
pegylated PLA immunonanoparticles (phi)[32]
6 V d v thit k mt h dn truyn thuc vo no in hnh
6.1 Khi nim Dendrimer
Cu trc ha hc ca dendrimer bao gm mt li ( cu trc trung tm ) v nhiu
nhnh to thnh mt cu trc hnh cu ba chiu gm cc lp ng tm. Do , cc cu
trc dendrimeric c c trng bi cc lp gia mi tiu im (hoc im cascade) gi
l bc (hnh 29). Nh vy, dendrimer bc 4 l cu trc hnh cu ng tm c 4 tiu im
gia cc li v b mt. Li ca dendrimer l bc khng bi v n khng c tiu im. Cu
trc dendrimer c hai tnh cht quan trng cho qu trnh dn truyn thuc. Th nht l
bn trong hc ca n c c tnh ha hc cho php bao bc cc loi thuc c tnh cht k
nc. Tnh nng quan trng th hai l cc dendrimer cha nhm chc b mt c trng

H nano dn truyn thuc cho no

Trang 34

GVHD: PGS.TS L Th Hng Nhan

cho php gn cc loi thuc a nc bng lin kt ha tr, tng tc tnh in hoc lin
kt hydro [33].

Hnh 29: Cu trc Denrimer

6.2 t vn .
S tng sinh mch mu v gia tng kch thc khng gii hn l nhng c tnh
ca khi u c tnh. p ng nhu cu cc cht dinh dng v oxy cho t bo khi u sinh
sn , cc khi u phi kch thch s hnh thnh cc mch mu mi thng qua cc qu trnh
tng sinh mc mu c kim sot ch yu bi VEGF . VEGF c nng qu mc bnh
thng v tit ra ch yu bi cc t bo khi u, c th kch thch s tng sinh ca cc t
bo ni m, gy hnh thnh mch trong m khi u. Do , cc h dn truyn thuc mi
cho no tp trung nghin cu cc phng php iu tr kp, c ngha l tiu dit t bo
ung th ng thi kim sot VEGF khng cho hnh thnh cc t bo ng th mi [34].

H nano dn truyn thuc cho no

Trang 35

GVHD: PGS.TS L Th Hng Nhan

Phng php can thip bng RNA (RNAi) l mt phng php mi y tim nng
iu tr cc khi u no, c th c ch chn lc cc gen mc tiu v ci thin nhy
ca liu php iu tr. Cc loi anti-VEGF c tng hp thnh cng v c kh nng
c ch hot ng ca VEGF lm mt mch mu gim v tng trng khi u chm hn
ng k. Mt mnh tc nhn anti-VEGF c th khng gy c ch hon ton tng
trng khi u, nhng c th lm tng s xm nhp ca hp thuc vo khi u bng cch
gim p lc ni bo v khi phc bnh thng chc nng mng li mao mch. t
c hiu qu iu tr tt, cc h dn truyn kt hp tc nhn c ch anti-VEGF v
doxorubicin (DOX) c s dng rng ri nhm lm tng hiu qu tiu dit cc t bo
khi u v ngn chn qu trnh to thnh cc khi u mi [34].
Tc nhn c ch VEGF c s dng trong nghin cu ny l shVEGF. i vi
cc liu php kt hp gia gen v tc nhn iu tr, s phn phi ng thi rt cn thit
trnh vic s dng liu lng cao .DOX c th xen vo chui xon kp ca DNA
to thnh dng phc plasmid-DOX , shVEGF- DOX. S an xen nh vy c th bo v
cc plasmid chng li nuclease v ci thin dc c tnh ng hc ca DOX bng cch
tng cng qu trnh lu thng mu. iu ny s c li cho kh nng c ch VEGF ca
shVEGF . Trong khi , DOX c th tiu dit t bo ung th mt cch hiu qu hn. T
nhng u im trn , shVEGF-DOX c th lm cho liu php kt hp v phn phi ng
thi thc hin d hn. iu ny s m ra nhng ng dng mi cho phng php ny
trong tng lai [34].
6.3 Phng thc tng hp v c ch dn truyn
Dendrigraft poly-L-lysine (DGL) c s dng rng ri cho qu trnh chuyn gen
vo khi u bi v chng c kh nng to phc hiu qu v bao bc tt DNA to thnh cc
ht nano. Trong cu trc, dtACPP c lin kt vi cc b mt DGL thng qua
Polyetylenglycol R-malemidyl--N-hydroxysuccinimidyl (MAL-PEG-NHS) to nn
h thng chuyn gen (gene nanocarrier) dtACPP-PEG-DGL (dtACPPD).Cui cng, cc
ht nano dtACPPD / shVEGFDOX c to thnh bng tng tc tnh in gia cation
DGL v plasmid shVEGF in tch m (hnh 30)[34].

H nano dn truyn thuc cho no

Trang 36

GVHD: PGS.TS L Th Hng Nhan

Hnh 30: Qa trnh tng hp dtACPPD / shVEGFDOX

H dn truyn nano ny c xu hng tch ly trong cc khi u bng cch ko di
thi gian tun hon v tng cng hiu ng th ng (EPR). Sau , cc ht nano c
ni bo v gii phng cc tc nhn iu tr. Cell-penetrating peptide (CPP) c s
dng rng ri dn truyn qua hng ro mu no .Tuy nhin , tnh c hiu v chn lc
ca phng php ny khng cao v CPPs c th tng tc v lin kt vi bt k thnh
phn no c mng t bo tch in m trong c th. khc phc nhc im ny,ngi
ta lm bt hot tnh cht CPP trong khi lu thng ngoi bo v khi phc cc c tnh ca
n trong mi trng ni bo no . Trong dtACPP, chc nng thc hin qu trnh ni bo
ca CPP polycationic c bt hot bng cch lin kt vi pH-sensitive masking
peptide thng qua cht nn matrix metalloproteinase 2 (MMP2). Trong qu trnh vn
chuyn, cc peptide masking (im ng in khong 6,4) s tch in m che chn tt
cho CPP. Trong khi u, s phn tch mi lin kt gia CPP polycationic v pH-sensitive
masking peptide bng MMP2 xy ra, CPP s c kch hot v vt qua hng ro mu

H nano dn truyn thuc cho no

Trang 37

GVHD: PGS.TS L Th Hng Nhan

no vo t bo theo con ng hp th qua trung gian transcytosis (Absorptive-mediated
transcytosis) [34].

Hnh 31: C ch dn truyn ca dtACPPD/shVEGF-DOX.

6.4 Kt qu thc nghim
Kch thc ht l mt tnh cht quan trng ca h nano c nh hng trc tip n
kh nng dn truyn thuc, s phn b kch thc ng knh ht thng c kim sot
di 200 nm. Trong nghin cu ny, kch thc h nano tng hp c c ng knh
di 150 nm to iu kin thun li cho vic vn chuyn thuc qua hng ro mu no
(hnh 32). Kch thc ca cc ht nano trong hnh SEM nh hn mt cht so vi cc kt
qu thu c t phng php nh sng tn x nguyn nhn l do s co li ca lp v cc
ht nano trong qu trnh sy chn khng qu trnh chun b mu SEM [34].

H nano dn truyn thuc cho no

Trang 38

GVHD: PGS.TS L Th Hng Nhan

Hnh 32: Ph phn b kch thc ht (DLS) ca dtACPPD/shVEGF-DOX pH=7.4

(tri) v pH=6( phi)
Huang v cc ng s cng kho st kh nng vn chuyn qua hng ro mu no
bng cch dng pht x hunh quang kim tra lng doxorubicin (DOX) trong BCECs.
Da vo cng pht x, ta thy dtACPPD / shVEGF-DOX c kh nng dn truyn
vo no tt nht (hnh 33) [34].

Hnh 33: Kho st kh nng dn truyn vo no ca cc h khc nhau qua pht x hunh
quang

H nano dn truyn thuc cho no

Trang 39

GVHD: PGS.TS L Th Hng Nhan

Kt qu thc nghim cho thy kh nng tch ly thuc dtACPPD / shVEGF-DOX
trong no tt hn cc h cn li.Thi gian sng trung bnh ca dtACPPD / shVEGF,
dtACPPD / control DOX v dtACPPD / shVEGF-DOX ln lt l 38, 35 v 58,5 ngy.
Thi gian sng st ca chut mang khi u no iu tr bng free DOX, CPPD/shVGEF,
dtACPPD / shVEGF v dtACPPD / control DOX khng di ln lt l 28, 32, 47 v 43
ngy. Ngc li, dtACPPD / shVEGF-DOX ko di ng k thi gian sng st ca
nhng con chut mang khi u ln ti hn 80 ngy (p<0.01).Cc kt qu trn cho thy
dtACPPD / shVEGF-DOX l h dn truyn nano tim nng cho no (hnh 34) [34].

Hnh 34: Kho st thi gian sng ca chut mang khi u no trong th nghim khi iu
tr bng cc h khc nhau
7 Kt lun
Nhng thp k gn y chng kin nhng tin b vt bc trong vic pht trin
cc h dn truyn thuc n no.Cc h nano cho thy kh nng ng dng tuyt vi
thit k cc h dn truyn nh hng mc tiu vt qua hng ro mu no , m ra mt
phm vi rng pht trin cc loi thuc da trn h nano .Tnh cht ha hc ca cc h

H nano dn truyn thuc cho no

Trang 40

GVHD: PGS.TS L Th Hng Nhan

dn truyn thuc "thng minh" to iu kin cho s phn phi v tng tc ca thuc vi
cc m c tnh mc tiu [35]. Chng lin tc c ci tin c th vn chuyn nhiu tc
nhn iu tr k c cc gene v bo v thuc khng b phn hy, ngn nga cc tc dng
ph v tng sinh kh dng thuc. Cc h dn truyn thuc "thng minh" hin nay c
s dng cho nhiu liu php iu tr, bao gm c ung th, nhim trng do vi khun,
khng virus, vaccine v tr liu gen, nhng ng dng to ln nht c l l iu tr cc bnh
lin quan n no[36]. Trong tng lai, hu ht phng php iu tr bnh no c th
c thc hin thng qua h nano. Tuy nhin,cc cng ngh phn phi thuc da trn ht
nano ang c hin nay vn cn phi c ci thin hn na chng c th an ton hn,
hiu qu hn, nh hng tt hn v hiu qu chi ph cao hn. Trc khi i vo cc th
nghim lm sng v c ng dng rng ri trong tng lai vn cn mt chng ng di
ti u ha, tiu chun ha cc liu php ny, nhng nhng kt qu thu c ban u
rt tch cc.

H nano dn truyn thuc cho no

Trang 41

GVHD: PGS.TS L Th Hng Nhan

Ti liu tham kho

[1]
Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM. Int J Cancer.
2010,127, 2893-917.
[2]
Stupp R, Mason WP, vanden Bent MJ, Weller M, Fisher B, Taphoorn MJB. New
Engl J Med. 2005,352, 987-96.
[3]

Sanai N, Berger MS. Neurosurgery .2008,62,753-64.

[4]
de Vries, N. A.; Beijnen, J. H.; Boogerd, W. van Tellingen. Neurother, 2006,6,
11991209.
[5]

Daneman R. Ann Neurol. 2012,72, 648-72.

[6]
Wang, B., Siahaan, T. Soltero, R.A. In: Drug Delivery: principles and
applications, Ed. Wiley, Inc., 2005.
[7]
Sampath Kumar, K. P.; Bhowmik, D.; Chiranjib; Chandira, M.;Tripathi, K. K. J.
Chem. Pharm. Res. 2010,2, 349-360.
[8]

Verma, R.K.; Mishra, B.; Garg, S. Drug Dev. Ind. Pharm. 2000,26, 695-708.

[9]
Ravikumar Reddy J.; Veera, M.; Mohamed T. S.; Madhu C. S. J.Pharm. Sci. & Res.
2009,1, 109-115.
[10] Kost, J.; Horbett, T.A.; Ratner, B.D.; Singh, M. J. Biomed. Mater.Res. 1985,19, 11171133.
[11]

de Boer AG, Gaillard PJ. Ann Rev Pharmacol Toxicol. 2007,47, 323-55.

[12] Wong AD, Ye M, Levy AF, Rothstein JD, Bergles DE, Searson PC. Front Neuroeng.
2013, 6, 7-12.
[13]

Pardridge WM. Pharm Res. 2007, 24, 1733-44.

[14]

Herv F, Ghinea N, Scherrmann JM. AAPS J. 2008,10, 455-72.

[15]

Lu W, Tan YZ, Hu KL, Jiang XG. Int J Pharm. 2005,295, 247-60.

[16]

Gupta B, Levchenko TS, Torchilin VP. Adv Drug Deliver Rev. 2005,57, 637-51.

[17]

Liu LH, Venkatraman SS, Yang YY, Guo K, Lu J, He BP. Pept Sci, 2008,90, 617-23.

[18]

Umezawa F, EtoY. Bio chem Biophs Res Commun, 1988,153, 1038-44.

H nano dn truyn thuc cho no

Trang 42

GVHD: PGS.TS L Th Hng Nhan

[19]

Qin Y, Fan W, Chen HL, Yao N, Tang WW, Tang J. J Drug Target. 2010,18, 536-

549.
[20]

Ponka P, Lok CN. Int J Biochem Cell B. 1999,31, 1111-37.

Zhang PC, Hu LJ, Yin Q, Zhang ZW, Feng LY, Li YP. J Control Release.
2012,159, 429-34.
[21]

[22]

Bromberg L.E., Ron E.S. Adv. Drug Deliv. Rev. 1998,31, 197-221.

[23]

Schild H.G. Prog. Polym. Sci. 1992,17, 163-249.

[24]

. Firestone B.A, Siegel R.A. Polym. Sci. 1991, 43, 901-914.

[25]

L. Brannon-Peppas, Peppas N.A.. Biomaterials, 1990,11, 635-644.

[26]

N.A. Peppas, J. Klier. J. Controlled Release , 1991,16, 203-214.

A. Mamada, T. Tanaka, D. Kungwachakun, M. Irie. Macromolecules, 1990,23,

1517-1519.
[27]

[28]

A. Suzuki, T. Tanaka. J. Appl. Phys, 1996,80, 131- 136.

[29]

Matsumura.Y.; Maeda. H. Cancer Res, 1986,46, 6387-6392.

[30]

Vicent. M. J.; Duncan.R. Trends Biotechnol. 2006,24, 39-47.

K. Kataoka, A. Harada,H. Nagasaki. Advanced Drug Delivery Reviews , 2001,47,

113131.
[31]

[32] Jean-Christophe O. The Journal of the American Society for Experimental

NeuroTherapeutics , 2005,2, 108-119.
[33] Quintana, A.; Raczka, E.; Piehler, L.; Lee, I.; Myc, A.; Mahoros, I.; Patri, A. K.; Thomas,
T.; Mul, J.; Baker. J. R. Pharm. Res., 2002,19, 1310-1316.
[34] S. Huang, K. Shao, Y. Liu, Y. Kuang, J.Li, S.An, Y. Guo, H. Ma, C. Jiang. Nature ,
2013,26, 119-128.
[35]

Demeule M, Currie JC, Bertrand Y, Ch C, Nguyen T, Rgina A. J Neurochem

2008, 106, 153444.

[36]

Xin HL, Jiang XY, Gu JJ, Sha XY, Chen LC, Law K. Biomaterials , 2011,32, 4293305.

H nano dn truyn thuc cho no

Trang 43