B Y T TRNG I HC DC H NI NG TH VNNH GI HIU LC IN VITRO CA 5 DN
XUT ARTEMISININ TRN CHNG PLASMODIUM FALCIPARUM KHNG CHLOROQUIN V TH
C TNH CP CA DN XUT C HIU LC CAO NHT TRN NG VT THC NGHIMKHA LUN TT
NGHIP DC S H NI - 2013B Y T TRNG I HC DC H NI NG TH VNNH GI HIU LC
IN VITRO CA 5 DN XUT ARTEMISININ TRN CHNG PLASMODIUM FALCIPARUM
KHNG CHLOROQUIN V TH C TNH CP CA DN XUT C HIU LC CAO NHT TRN NG VT
THC NGHIMKHA LUN TT NGHIP DC S Ngi hng dn : 1. Ths. L Th Thu Hng 2.
TS. Bi Quang PhcNi thc hin : Khoa Nghin cu iu tr st rt Vin St rt K
sinh trng -Cn trng Trung ngH NI - 2013LI CM Nthc hin v hon thnh kha
lun tt nghip ny, li u tin ti xin by t lng knh
trngvbitnsuscticgioThS.LThThuHng(BmnVisinhSinhhc, trngihcDcH Ni)
ngi thy trc tip hng dn, ch bo tn tnh v cho ti
nhnglikhuynqubutrongsutqutrnhnghincuvhonthnhkhalun tt nghip
ny.TixinbytlngbitnchnthnhtithyTS.BiQuangPhc(KhoaNghincu iu tr st
rt, Vin St rt K sinh trng Cn trng Trung ng) cng tp th cn b trong
Khoa c bit ch TS. Nguyn Th Minh Thu hng dn tn tnh, to mi iu kin
thun li gip ti trong qu trnh thc nghim v nghin cu hon thnh kha lun
ny.TicngxinchnthnhcmnccthyctrongBanGimhiu,Phngoto,cc phngbanv cc b
mn Trng i Hc Dc H Ni dy d, to iu kin trong 5 nm hc v hon thnh tt
kha hc.Cui cng, ti xin gi n nhng ngi thn trong gia nh, bn b c bit
nhmNNCC, nhng ngi lun bn cnhng vin, gip ti li cm n su sc trong sut
qu trnh hc tp v hon thnh kha lun tt nghip.H Ni, ngy 14 thng 5 nm
2013Sinh vinNg Th VnMC LCDANH MC CC K HIU, CC CH VIT TTDANH MC CC
BNGDANH MC CC HNH V, BIU , S T VN
..........................................................................................................................
1CHNG 1. TNG
QUAN..................................................................................................
31.1. K SINH TRNG ST RT
PLASMODIUM..........................................................
31.1.1. c im hnh
thi........................................................................................................
31.1.2. Chu k pht
trin...........................................................................................................
41.1.2.1. Chu k pht trin v tnh trong ngi
...................................................................
41.1.2.2. Chu k pht trin hu tnh trong c th
mui...................................................... 51.2. TNH
HNH ST RT TRN TH GII V VIT NAM...................................
51.2.1. Vi nt khi qut v bnh st
rt................................................................................
51.2.2. Tnh hnh bnh st rt trn th gii v Vit
Nam................................................... 61.2.2.1.
Tnh hnh bnh st rt trn th
gii........................................................................
61.2.2.2. Tnh hnh bnh st rt Vit
Nam........................................................................
61.2.3. Tnh hnh khng thuc ca
Plasmodium..................................................................
71.2.3.1. Tnh hnh khng thuc ca Plasmodium trn th
gii........................................ 71.2.3.2. Tnh hnh khng
thuc ca Plasmodium Vit Nam.........................................
81.3. C IM ARTEMISININ V DN
XUT..........................................................
91.3.1. Ngun gc, tnh cht ca artemisinin v dn
xut.................................................. 91.3.1.1.
Ngun gc, cu trc, tnh cht ca artemisinin
................................................... 91.3.1.2. c im
mt s dn xut ca
artemisinin...........................................................
91.3.2. C ch tc dng v nhc im chng st rt ca ART v dn xut
............... 101.3.2.1. C ch tc dng ca artemisinin v dn
xut..................................................... 101.3.2.2.
Nhc im ca artemisinin v dn
xut............................................................
111.4. TNH HNH NGHIN CU 5 DN XUT ARTEMISININ 32, 33, 39c, 39d, 39e
121.4.1. Tnh hnh nghin cu, u im ca cc dn xut 10
deoxoartemisinin........ 121.4.2. Cu trc, tnh cht ca 5 dn xut 32,
33, 39c, 39d, 39e ....................................
121.4.2.1.Nghincutnghp10-[(2-hydroxy-3-imidazol)propyl]deoxoartemisinin(32)v
10-[(2-hydroxy-3-imidazol) propyl] deoxoartemisinin
(33).....................................
131.4.2.2.Nghincutnghpccdnxutamid10-(deoxoartemisinin)-N-((pyridin-3-yl)
methyl)acetamide(39c),10-(deoxoartemisinin)-N-((pyridin-2-yl)methyl)acetamide(39d),10-(deoxoartemisinin)-N-phenethylacetamide
(39e).............................................................
131.4.2.3. Tnh cht ca 5 dn xut
........................................................................................
14CHNG 2. I TNG V PHNG PHP NGHIN CU............................
162.1. NGUYN VT LIU, THIT B
..............................................................................
162.1.1. Chng k sinh trng st rt
.......................................................................................
162.1.2. ng vt th
nghim...................................................................................................
162.1.3. Thuc v ha cht nghin
cu..................................................................................
162.1.4. Dng c v thit b dng trong nghin
cu............................................................
172.2. NI DUNG NGHIN
CU........................................................................................
172.3. PHNG PHP NGHIN
CU...............................................................................
172.3.1. Th nghim in vitro ca 5 dn xut 32, 33, 39c, 39d, 39e trn
chng P.falciparum khng CHQ nui cy
.........................................................................................................................
172.3.1.1. Nui cy KSTSR P. falciparum lin tc trong phng th
nghim................. 172.3.1.2. Th nghim in vitro theo phng php
th thuc 48 gi................................ 192.3.2. Th nghim c
tnh cp ca dn xut c hot lc in vitro mnh nht ........... 20CHNG 3.
THC NGHIM, KT QU, BN
LUN............................................ 233.1. KT QU NGHIN
CU...........................................................................................
233.1.1. Th nghim hiu lc in vitro ca 5 dn xut 32, 33, 39c, 39d, 39e
................. 233.1.2. Th nghim c tnh cp ca hp cht
32.............................................................
323.2. BN
LUN.....................................................................................................................
333.2.1. Kt qu th hot lc in vitro ca 5 dn xut 32, 33, 39c, 39d, 39e
.................. 333.2.2. Kt qu nghin cu c tnh cp dn xut 32 cht c
hiu lc in vitro mnh nht 35KT LUN V KIN
NGH..............................................................................................
37KT
LUN.............................................................................................................................
37KIN
NGH.............................................................................................................................
38TI LIU THAM KHODANH MC CC K HIU, CH VIT TTTing vit32
10-[2-hydroxy-3-imidazol) propyl] deoxoartemisinin33
10-[2-hydroxy-3-imidazol) propyl]
deoxoartemisinin39c10-(deoxoartemisinin)-N-((pyridin-3-yl) methyl)
acetamide39d10-(deoxoartemisinin)-N-((pyridin-2-yl) methyl)
acetamide39e 10-(deoxoartemisinin)-N-phenethylacetamideAE
ArteetherAM ArtemetherART ArtemisininAS ArtesunatBN Bnh nhnBNSR Bnh
nhn st rtCHQ ChloroquinCMNng ca cc mu th tnh theo th tchDHA
DihydroartemisininDMSO Dimethyl sulfoxideVTN ng vt thc nghimHC Hng
cuKST K sinh trngKSTSR K sinh trng st rtMKST Mt k sinh trngP.
falciparum Plasmodium falciparumP. vivax Plasmodium vivaxRI, II,
III Mc khng thuc ca k sinh trng st rtSKD Sinh kh dngSR St rtTCYTTG
T chc y t th giiTLPT Trng lng phn tTing anhGHSGlobally Harmonised
Classification System for Chemical Substances and MixturesH thng
phn loi c tnh cp theo gi tr gii hn LD50IC50Inhibitory Concentration
50%Nng c ch 50% k sinh trng pht trinLD50Lethal Dose 50Liu cht 50%
ng vt th nghimMIC Minimum Inhibitory ConcentrationNng ti thiu c ch
k sinh trng pht trinWHOWorld Health Organization T chc y t th
giiDANH MC CC HNH V, BIU Biu Tn biu Trang3.1 T l c ch chng P.
falciparum K1 ca cht 32 253.2 Gi tr IC50 ca ART v 5 dn xut trn chng
P. falciparum K1 303.3 Gi tr MIC ca ART v 5 dn xut trn chng P.
falciparum K1 31Hnh Tn hnh Trang1.1 Cng thc cu to ca ART 91.2 S bin
i ca dn xut ART trong huyt tng 111.3 Qui trnh tng hp cc cht 32 v 33
131.4 Qui trnh tng hp cc dn xut amid ca ART (39a-e) 14DANH MC CC
BNGBng Tn bng Trang1.1 c trng l ha c bn ca 5 dn xut
153.1MtKSTchngP.falciparumkhngCHQtrong10ngy trc khi th thuc233.2S p
ng ca chng P. falciparum K1 trn in vitro vi cht 32243.3S p ng ca
chng P. falciparum K1 trn in vitro vi cht
33263.4SpngcachngP.falciparumK1trninvitrovicht
39c273.5SpngcachngP.falciparumK1trninvitrovicht
39d283.6SpngcachngP.falciparumK1trninvitrovicht 39e293.7Nng c ch
50% KST pht trin ca 5 dn xut chng
K1293.8Nngcchtithiuca5dnxuttrnchng P.falciparum K1313.9Kt qu sau 7
ngy theo di chut ung cht 32 nng khc
nhau323.10Ktququanstcnnngvithcccquankhimton b c 13 l chut sau 7 ngy
theo di33T VN
BnhStrt(SR)lmtbnhlytheongmu,doksinhtrng(KST)Plasmodiumc truyn t ngi
bnh sang ngi lnh bi mui thuc ging Anopheles. Y t th gii (WHO) nm
2012 cn 99 quc gia st rt lu hnh nng v s ca mc st rt c tnh chim 78%
ton cu. Vit Nam l khu vc st rt lu hnh nng, c bit l khu vc trung
Namb v ng Nam b. Trong nhng nm gn y st rt ang tng ln, ch trong nm
2011 c 15,6 triu ngi nhim k sinh trng st rt v 10% trong b t vong
[41].Tnhtrngxuthinvlanrngksinhtrngstrtkhngthucangedamts chng trnh
phng chng st rt, c bit l k sinh trng Plasmodium falciparum khng li
cloroquin v cc thuc truyn thng khc. T nm 1990, thuc artemisinin v
dn cht artesunat c tin hnh th nghim lm sng v chnh thc a vo s dng
rng ri trong Chng trnh Quc gia phng chng st rt nc ta gp phn quan
trng trong trong vic h thp t l bnh vtltvongdostrt.Tuy nhin nhc im
ca artemisinin v dn xut l thi gian bn
thingnvtltiphtcao[4].Ngoira,theobococaWHOnm2012Plasmodium
falciparum khng li artemisinin nhiu khu vc nh ng Nam , Chu Phi c
bit l cc nckhuvcsngM-kng.Do,victmrathuciutrmichiulccaovi Plasmodium
khng thuc l mc tiu cp bch ca cc nh khoa hc trn ton th gii
[41].Hinnay,hngnghincutnghpccdnxutartemisininchonhiuktqutt,
nibtldnxutmonohocdimecaartemisinincchalinkt10-C-Cbnvng.it
dihydroartemisininvsnphmtrunggian10-allyldeoxoartemisinin,ccnhkhoahcca
VinHahclnutinsnxutthnhcng5dnxutcaartemisininl:32,33,39c, 39d, 39e.
Cc hp cht ny bc u th hin hot tnh chng st rt in vitro v in vivo vt
tri so vi artemisinin v arteether [36], [37].
Vvy,gpphnnhgihiulccaccdnxutartemisinintrnPlasmodium
falciparumchngtitinhnhtinhgi hiu lc in vitro ca 5 dn xut
artemisinin
trnchngPlasmodiumfalciparumkhngcloroquinvthctnhcpcadnxutc hiu lc
cao nht trn ng vt thc nghim vi cc mc tiu sau:Nghin cu hiu lc in
vitro ca 5 dn xut artemisinin 32, 33, 39c, 39d, 39e trn chng
Plasmodium falciparum khng cloroquin.Th nghim c tnh cp ca dn xut c
hiu lc mnh nht trn ng vt thc nghim.CHNG 1. TNG QUAN1.1. K SINH TRNG
ST RT PLASMODIUM1.1.1. c im hnh thi Trn Tri t c ti 120 loi
Plasmodium gy bnh trn ng vt nhng ch c 4 loi gy
bnhchongi:Plasmodiumfalciparum(P.falciparum),Plasmodiumvivax(P.vivax),
Plasmodiumovale(P.ovale),Plasmodiummalariae
(P.malariae)[8],[10].Cnhhngcu
(HC)trnphinknh(lamknh)rinhumtheophngphpGiemsahayccphngphp tng t
(Wright, Field) c th quan st v nh loi k sinh trng st rt (KSTSR).
Trn tiu bn thng thy KSTSR gm 2 thnh phn chnh l nguyn sinh cht bt mu
xanh v nhn bt
mu,ngoiracnthycchtsct[8],[15].TronggiaionHCngi,KSTSRc3 dng hnh th
[8], [15]: - Th t dng (Trophozoite hay th nhn): th t dng non c hnh
nhn, ng knh khong 1/5 1/3 ng knh HC, gm mt nhn mu , t bo cht mu
xanh da tri v mt khng bo to, chim phn ln t bo cht. Th t dng gi c
hnh dng ging amip.- Th phn lit (Schizonte): Nhn v t bo cht phn
chia. Th phn lit gi, mi mnh nhn
cmtmnhtbochtbaoquanh(mnhtrng/merozoite).Slngmnhtrngtyvo loi
Plasmmodium.- Th giao bo (Gametocyte): L th hu tnh, hnh dng khc
nhau ty theo loi.* Hnh th ca P. falciparum trn lam mu nhum theo
phng php Giemsa [8],
[15]:-Thtdngnon:Nhnnhcngknhchbng1/51/6ngknhHC.Thnhn thng nm ra t bo
HC v hay gp nhn c 2 nhn.- Th t dng gi: Nguyn sinh cht dy hn, nhn to
ra, khng bo bin mt v xut hin cc htsct hnh que, mu nu nh vng, ri rc
hoc tp trung thnh tngm trn nguyn sinh cht. Th ny t thy mu ngoi
vi.-Thphnlit:Nhnvnguynsinhchtphnchiathnh832mnh.Khichnmui,mi mnh nhn
c 1 vng nguyn sinh cht bao quanh to thnh 8 32 mnh trng. Rt him thy
mu ngoi vi, thng ch thy th SR nng hoc SR c tnh, bnh thng chng tp
trung trong mu mao mch.- Th giao bo: Giao bo c hnh qu thn, u trn,
nguyn sinh cht c mu tm hoa c, nhn
khngcgiihnrrt.Nhnghtsctmunu,th,t,phnbrirctrong nguyn sinh
cht,xenkcnhnghtsctmuen.Giaobocihnhquchuihaylilim,nguyn sinh cht c
mu xanh da tri, nhn bt mu tp trung gia, xung quanh l nhng ht sc t
mu en [8], [15].1.1.2. Chu k pht trinChu k ca KSTSR gm 2 giai on
pht trin rt phc tp gm 1 chu k pht trin v tnh trong ngi v 1 chu k
sinh sn hu tnh trong mui Anopheles.1.1.2.1. Chu k pht trin v tnh
trong ngi a) Giai on tin hng cu (Giai on trong
gan)Khimuitngi,thoatrngctrongtuynncbtcamuiSRthucging Anopheles
truyn vo ngi, xm nhp vo mu, ch tn ti trong mu ngoi vi khong 30 pht.
Sau , mt b phn (thot khi tc ng ca min dch t bo) xm nhp vo t bo gan,
pht trin ri phn chia thnh th phn lit. Th phn lit pht trin y s ph v
t bo gan, gii
phngrartnhiumnhtrngcagan,ccmnhtrngnysivomuvksinhtrongt bo HC [4],
[8], [15].i vi P. falciparum v P. malariae: Tt c cc thoa trng vo
gan u pht trin v trng thnh, phn chia thnh cc mnh trng ri vo mu
trong thi gian tng i ngn [8], [15].i vi P. vivax v P. ovale, ngoi s
pht trin tc th ca a s thoa trng, c mt s thoa
trngkhngphttrinngaymtothnhth ng li t bo gan, c th sau 1, 2, 3 6
thng mi pht trin mun hn thnh th n gy cc cn SR ti pht xa giai on
ngoi HC [8],
[15].ThigianhonthnhgiaiontinHC:P.falciparuml5-7ngy,P.malariael14-16
ngy, P. vivax l 7-8 ngy, P. ovale l 9-10 ngy [8], [15].b) Giai on
pht trin trong hng cu (Giai on trong mu)Trong mu, cc mnh trng c gii
phng t t bo gan s xm nhp vo HC, pht trin 1chuksinhsnvtnh.Ln lt th t
dng non, ri pht trin thnh th t dng gi,saunhnvnguynsinh cht c phn
chia ra lm nhiu mnh to thnh th phn
lit.ThphnlitphttrinysphvHC,giiphngra cc mnh trng ca HC. Hu
htccmnhtrngnylitip tc xm nhp vo cc HC bnh thng mi sinh sn v tnh
trong HC. Mt s mnh trng bit ha thnh th hu tnh: cc giao bo c v giao
bo ci. Nu khng c mui truyn bnh ht vo d dy, cc giao bo ny s t tiu hy
trong mu [4], [8],
[15].ThigianhonthnhmtchukcaKSTSRtrongHCkhcnhautyloiKSTSR:P.
falciparumkhong48gi,gystcchnhthocsthngngynunhimnhiudngkhc nhau; P.
vivax v P. ovale khong 48 gi, gy st cch nht; P. malariae khong 72
gi gy st 3 ngy mt cn [8], [15].1.1.2.2. Chu k pht trin hu tnh trong
c th muiSau khi c mui Anopheles ht vo d dy, cc giao bo c v ci pht
trin thnh giao t. Giao t c v ci phi hp vi nhau to thnh hp t ri trng
di ng. Trng di ng chui
quathnhddymui,phttrintrnmtngoicathnhddy,trnli,todn,phnchia
thnhnhiuthoatrngbntrong.Cuicng,thoatrngcgiiphng,dichuynvtuyn nc bt
ca mui, khi mui t ngi thoa trng xm nhp c th ngi v bt u chu k sinh
sn v tnh trong ngi [4], [8], [15].1.2. TNH HNH ST RT TRN TH GII V
VIT NAM1.2.1. Vi nt khi qut v bnh st rt
BnhstrtlbnhlynhimdoksinhtrngnbothucgingPlasmodium gy ra [10]. Lm
sng bnh SR thng c phn ra lm hai loi l SR th thng thng v SR c tnh.
ng thi cn c mt s th SR c bit khc: SR tr em, SR ph n c thai, SR bm
sinh, SR do truyn mu, SR mn tnh, SR ni tng tin trin [8]. Bnh biu
hin in hnh bng nhng cn st rt vi 3 triu chng c bn: rt run, st, ra m
hi [15]. Tn s v cc mc cc cn st rt ty thuc vo loi KSTSR, nhng thng
thng l 2-3 ngy. Khi cc t k sinh trng ng thi nhn ln, hng cu b ph hy
hoc c th kt dnh vo nhau v vi thnh mch mu xuthintvong cc t bo ny lm
tc nghn mch mu cung cp mu cho no (SR th no) v cc t chc khc [10],
[39].1.2.2. Tnh hnh bnh st rt trn th gii v Vit Nam 1.2.2.1. Tnh hnh
bnh st rt trn th gii Bnh st rt c mt, lu hnh rng khp vng nhit i, vng
cn nhit i v cc vng kh hu n ha [10]. Bnh SR lan truyn vng cn sa mc
Sahara, Chu Phi c mt hu ht mini,ncc khu vc ring bit khng ng u nh ng
Nam , n , Trung v Nam M [4].Bnh SR vn l gnh nng sc khe ton cu vi gn
mt na dn s trn th gii c nguy c mc st rt, mi nm c khong 250 triu ca
mc trong 1 triu ca t vong [28]. Trong chu Phi chim 90% v c cho l
khu vc c SR lu hnh nng vi gn 1 triu tr em di 5
tuihngnmchtdostrtnthunhocphihpviccbnhkhc.Tnhtrungbnhc 125 a tr cht
trong 1 gi v 2 a tr cht trong 1 pht [4]. Sau chu Phi, khu vc chu
Thi Bnh Dng l ni b nh hng nng n vi 21 triu ngi mc st rt hng nm
[34]. Ti ccnckhuvcngNam,mcdtrongnhngnmgny,tlmcstrtxcnh
trongkhuvcgimxpx75%t2000n2011[41],nhngbnhSRvnangluhnh vi hu ht cc
nc vi 88% dn s trong tng s 1320 triu ngi sng trong khu vc [4].
1.2.2.2. Tnh hnh bnh st rt Vit Nam
BnhSRVitNamchyudoP.falciparumgyra(lloithnggySRctnh) chim 80-85%, P.
vivax chim 15-20%, P. malariae chim 1-2% [23].Vit Nam l quc gia c
khong 20% dn s sng trong vng st rt lu hnh, do iu kin kh hu nhit i
gi ma, nhit v m rt thch hp cho bnh st rt lu hnh nhiu a phng khu vc
cc tnh min ni pha Bc, cc tnh ven bin min Trung, khu vc Ty Nguyn v
ng Nam B [4], [17], [41].T nm 1958, chng trnh thanh ton st rt c thc
hin min Bc v m rng ra c ncnm1976thuc nhiu kt qu tt. Nhng cui nhng
nm 70 80 do thiu ngun
lc,dibinngdnc,huquchintranhvnhiunguynnhnkhcnnbnhstrt quay tr li v
ngy cng nghim trng. Nm 1991 Vit Nam chuyn sang chin lc PCSR do
WHOxngcktqunitritheonhgigiaion20012005bnhnhnstrt (BNSR) gim 69%,
cht do st rt gim 87%
[2].TheobococaWHOnm2012tlmcstrtVitNamgimtrn75%,iu
nychngtbnhSRcytvcngngrtquantmcbitvngsu,vngxa, vng dn tc thiu s, ni
b con cn cha c nhiu kin thc v bnh SR [41].1.2.3. Tnh hnh khng thuc
ca Plasmodium1.2.3.1. Tnh hnh khng thuc ca Plasmodium trn th gii
Trongnhngnmgny,khngthucangl thch thc ln trong phng chng st rt trn
ton cu, c bit l chng Plasmodium falciparum c pht hin t u th k 20.Nm
1910 P. falciparum c pht hin khng quinin u tin Brazil v hin khng
nhiuvngngNam,NamM,ChuPhi(Tanzania)[16],[40].Nm1949,P. falciparumv
P. malariae khng proguanin c pht hin Brazil v sau lan ra Kennya,
Tanzania, Chi
L,Malaysia,IndonesiavVitNam(Peter,1970)[16].Nm1957,trnghputinP.
falciparumkhngchloroquin(CHQ)phthinbin gii Colombia v Venezuela,
tip sau
lThiLan(Harinasuta,1961).TheothngkcaWHO,tnhtrngkhngCHQlanrng
khpthgiichim75/95qucgiakhngthuc,bncnh,hintngkhngCHQgp10 ln so vi 50
nm trc [16],
[20].Ngoira,cnphthinthmrtnhiutrnghpP.falciparumkhngthuckhcnh
pyrimethamin,piperaquin(PQ),FansidarcbithnnaltnhtrngP.falciparumkhng
li artemisinin (ART) v dn xut thuc c hiu lc hng u hin nay. Trong l
hin tng khng vi ART xy ra ti khu vc ng Nam , ng ch l vng sng M Kng
vi 4 nc: Lo, Campuchia, Thi Lan, Vit Nam [16], [20],
[41].Nhvy,hinnayKSTSRkhngvihuhtccloithucSRrtnhiunctrn thgii,y l tr
ngi rt ln cho cng tc phng chng SR [16]. Vn t ra hin nay cho
tonthgiiltmrathuchocdnxutchiulciutrcaovkhngbP.falciparumkhng.1.2.3.2.
Tnh hnh khng thuc ca Plasmodium Vit Nam Tnh trng P. falciparum khng
CHQ ln u tin ti Vit Nam c ghi nhn Nha Trang
nm1961,saulanracNamTrungB,TyNguyn,minNam,nhTy Nguyn (1964), ng Hi
(1967), Qung Tr (1973), cng trng 7 Qung Bnh (1967), Bnh nh (1968),
Ty Ninh (1964), Phan Thit (1974)...[11]. Hin nay, xu hng khng CHQ
ang tng tr li, nghin
cutiQungTrnm1999chothyP.falciparumcmckhngRIIviCHQl7,1%,ti Qung Trc,
tnh k Lk t l tht bi iu tr (RI, RII v RIII) ca phc CHQ l 48% [22],
nnm2008-2009tlkhngCHQQungTrl75%,tiknnglnn81%[7]. Fansidar ra i nm
1964 v nhanh chng b khng Vit Nam vi t l khng cao trn 50% RI, RII,
RIII (1986 1994)
[15].cbitlxuthintnhtrngP.falciparumkhngliartemisininvdnxut.Theo
nghincutitnhSngBvGiaLai(1996) ART dng vi tng liu 60 mg/kg trong 5
ngy th t l ti pht l 16,7%, ti min trung Ty Nguyn (1996) vi liu dng
1000 mg/kg trong 5
ngythtltiphtl34,5%,kcviphcphihpART+mefloquin,ART+ tetracyclin th t
l ti pht ln lt l 6,42%; 14,4% [24], [29]. Hiu lc iu tr artesunat vi
P. falciparumtuycaonhngtlthtbimuncncn nhc: ti Gia Lai (2001 2002)
vi liu dng ngy u 4 mg/kg, ngy 2-4 gim xung 2 mg/kg th TLTBM l
46,7%; Bnh Phc (k Nhau B ng) (2009) vi tng liu 16 mg/kg trong 7 ngy
th TLTBM l 14,6% [6], [12].Nhvyvicgimstsdngthucvtm ra hng dng thuc
mi l iu quan trng trong cng cuc chng li KSTSR khng thuc hin
nay.1.3. C IM ARTEMISININ V DN XUT1.3.1. Ngun gc, tnh cht ca
artemisinin v dn xut 1.3.1.1. Ngun gc, cu trc, tnh cht ca
artemisinin Artemisinin l mt hot cht chng SR c chit xut t cy Thanh
hao hoa vng vi tn
khoahclArtemisiaannuaL.,hccArteraceae[21].Nm1971cccnhkhoahc Trung
Quc chit xut thnh cng va vo s dng rng ri t nm 1972, Vit Nam ART bt
u a vo s dng iu tr cho khu vc c P. falciparum khng thuc t nm 1991
[4].ARTctinhthhnhkim,khngmu,imchy156-157C,cngthcphntl C15H22O11,
trng lng phn t (TLPT) l 282 v cu to nh hnh 1.1 [3], [21], [31].Hnh
1.1. Cng thc cu to ca ART [21], [31].ART l sesquiterpen lacton c cu
ni peroxid ni phn t v hot tnh chng SR ch yu da vo cu ni ny [21],
[31], [38]. Trong cng thc trn vng A l dng cycloxan, vng B l vng
lacton, cn vng C v D u l d vng bo ha trong vng D l mt trioxan, dng
lin quan n hot tnh chng SR ca ART [21].
ARTttantrongncvdu,rtdbthyphnvnglactontrongdungmiphncc [31], [38].
Do vy sinh kh dng (SKD) v bn vng ca ART khng cao, v y cng chnh l hn
ch ca ART khi dng [3]. 1.3.1.2. c im mt s dn xut ca artemisinin Vi
mc ch tng cng kh nng ha tan, tng hot lc chng SR, u tin nm 1976 cc
nh khoa hc Trung Quc tng hp thnh cng dihydroartemisinin (DHA) bng
cch kh ha
nhmCOlactonthnhOHbnacetalvitcnhnkhhalnatriborohydric[21].Mcd DHA c
hot lc mnh hn gp 2 ln ART nhng n vn t tan trong nc, du nn DHA c s
dng nh l 1 cht trung gian quan trng bn tng hp cc dn xut khc
[21].ThngquaphnngetherhaDHA,cccdnxutmiraivcngdng
nhiunhtlartemether(AM)varteether(AE)TLPTlnltl298v312,ctnhlha
tngtART,uimhnARTldtantrongdu,dungmihucnhethanol,aceton,
chloroform...[31],
[38].Artesunat(AS)lsnphmcaphnngesterhaDHA,cithinckhnngtan
trongnccaART,csdngrngriccncngNamvChuPhiditk sinh trng st rt P.
falciparum [14].Ngoira,cnnhiuphngphptnghpccdnxutcaARTtDHAangc nghin
cu hin nay c kh nng mang li nhiu hiu qu cao [21].1.3.2. C ch tc dng
v nhc im chng st rt ca ART v dn xut 1.3.2.1. C ch tc dng ca
Artemisinin v dn xut Qua nhiu nghin cu v c ch tc dng t khi chit xut
thnh cng ART nhng c ch tcdngcaARTvdnxutvncn nhiu tranh ci. N c th l
s phi hp nhiu c ch nh tc ng ln mng, c ch tng hp protein, gc t do
hay oxy hot ng
[21].TheoktqucanhmnghincuquinghaosuTrungQuc,ARTtcnglnmng ca KSTSR l
tc dng chnh.u tin l lm bin i cu trc mng, sau to ra cc khng bo t
tiu, cui cng lm tan r bn trong KST [35]. Theo Gu H.M v cng s cho
rng DHA v AM c kh nng c ch in vitro vi tng hp
proteinhngcunginhimP.falciparum,trongcchtnghpacidnucleicxyra chm hn
tng hp protein [20]. Do c th thy tc dng ca thuc khng tp trung vo qu
trnh tng hp acid nucleic v tc dng chn lc ca thuc i vi hng cu b nhim
[21].TheoMeshnickvcngs,stngvaitrquantrngtrongcuniperoxidenivs
tothnhgctdo. ART v dn xut sau khi vo mu tng tc vi hemozoin (sn phm
tiu ha hemoglobin ca KSTSR) sn sinh ra cc gc t do c carbon trung
tm. Cc gc t do ny gy tn hi t bo nh peroxy ha lipid, oxy ha protein,
alkyl ha protein v git cht KST.
TcdngnyrrtnhtthtdngcaKSTSR.McdtcdngditKST ca ART bt
usaukhitipxcchm(4-5gi)hnviCHQnhngtcdngcchttiartnhanh c bit vi P.
falciparum khng CHQ (do c hemozoin) [21], [30], [32], [33], [38].
1.3.2.2. Nhc im ca artemisinin v dn xut
ARTvccdnxutctcdngnibtlctstnhanh,schthvtnhcaKSTSR nhanh v k c th
giao bo [4], [20]. Nng IC50 ca ART t 10-8 10-7 mol/L (3 30 g/L),
nng thp nht c ch l 3 x 10-8 mol/L. Hot tnh ca DHA cao hn ART t 4 5
ln (IC50t 0,36 7 nmol/L), dn xut khc ca ART u c hot tnh cao hn ART
nh AS cao hn 4 5 ln, AM v AE hn khong 2 ln [38]. c bit ART v dn xut
c hiu qu tt trn KSTSR khng thuc hoc a khng thuc [4], [38].Nhng do c
im cu to c bit ca ART v dn xut nh kh nng tan trong nc v
trongdukm,nhncathuccao,cchalinkt10-hemiacetaldb chuyn ha thnh DHA
di s oxy ha bi men P-450 trong gan nn SKD ca thuc gim v thuc hp thu
chm [26], [37]. nh hng ca chuyn ha khin ART v dn xut c thi gian bn
thi ngn, v vy sau khi iu tr, bnh nhn vn cn t l ti pht li k sinh
trng v t l ti pht cao hay thp ty thuc vo thi gian dng thuc ngn hay
di (7 ngy t l ti pht vn cn 10%) [4]. Hnh 1.2. S bin i ca dn xut ART
trong huyt tng [37].Ngoi ra, theo nghin cu dc l c tnh trn thn kinh
ca ART v dn xut trn ng
vt,huhtccdnxutucctnh,nhngDHA(snphmchuynhacahuhtcc ete) th nh hng
nng n trn thn kinh trung ng [20].Nh vy, hin nay hng nghin cu ca cc
dn xut ART vi mc ch khc phc nhc
imcaARTcphttrintheo2hng:hngthnhtltoraccdnxutmiit
ARTvDHA,hngth2lphihp1dn xut ca ART vi 1 thuc SR khc c thi gian bn
thi di hn [20].1.4.TNHHNHNGHINCU5DNXUTARTEMISININ32,33, 39c, 39d,
39e 1.4.1. Tnh hnh nghin cu, u im ca cc dn xut 10 deoxoartemisinin
Tnh hnh khng thuc ngy cng gia tng i hi mt hng iu tr hoc thuc iu tr
mivvictnghpccdnxutmicaARTcccnhkhoahc trn th gii quan tm nghin cu, c
bit l thay i cu trc lacton ca DHA, to ra cc dn xut bn hn v c hot
tnh cao hn. V d nh cc dn xut fluoroartemisinin, 11-aza-artemisinin
[13].
Theoccnghincugnychothyccdnxutnon-acetalcaART,gil10-deoxoartemisinincbnvng15-22lnhnccdnxutacetaltrongiukinmphng
ca d dy [27]. Bn cnh dn xut mi 10-deoxoartemisinin vi d vng, amin v
nhm thn nc th hin hot tnh chng st rt in vitro cao hn 10 25 ln so vi
ART [36].iu ny cho thy nhng hp cht c b khung c bn ca ART nhng khng
cha nhm chc lacton, acetal m thay vo l cc dn xut cha lin kt 10-C-C,
cha nit c kh nng
tantttrongncvtrongdu,tinlichovicsdngnhkhnngtomuitanviacid nh tnh
baz ca nit ang l hng i mi v ng n trong vic phng chng st rt
[37].1.4.2. Cu trc, tnh cht ca 5 dn xut 32, 33, 39c, 39d, 39e Theo
hng nghin cu trn Vin Ha hc nghin cu tng hp thnh cng 5 dn xut t DHA
v sn phm trung gian 10-allyldeoxoartemisinin.1.4.2.1. Nghin cu tng
hp 10-[(2-hydroxy-3-imidazol) propyl] deoxoartemisinin (32) v
10-[(2-hydroxy-3-imidazol) propyl] deoxoartemisinin (33) Qui trnh
tng hp i t DHA (9) qua 4 bc phn ng (to 34, 35, 36) thu c cc cht 32,
33 theo hnh 1.3 [27], [36], [37].Hnh 1.3. Qui trnh tng hp cc cht 32
v 33 [27], [36], [37].Cht phn ng v iu kin phn ng :Benzoyl chloride,
pyridin, CH2Cl2, nhit phng, 16h, 97%.Allyltrimethylsilane, ZnCl2,
ray phn t, 1,2-dicloetan, 0C, 1h, 90%.m-CPBA, nhit phng, 24h,
CH2Cl2, 85%.Imidazol, C2H5OH/H2O, nhit phng,
20h.1.4.2.2.Nghincutnghpccdnxutamid10-(deoxoartemisinin)-N-((pyridin-3-yl)
methyl) acetamide (39c), 10-(deoxoartemisinin)-N-((pyridin-2-yl)
methyl) acetamide (39d), 10-(deoxoartemisinin)-N-phenethylacetamide
(39e) [37] Thpcht 35 c iu ch t DHA trn qua 2 bc to thnh cc dn xut
amid theo hnh 1.4 [37].Hnh 1.4. Qui trnh tng hp cc dn xut amid ca
ART (39a-e) [37].Saukhitng hp 5 dn xutu c nh gi kt qu thng qua ph
cng hng t ht nhn 1H NMR xc nh li cu trc hp cht
[37].Vy,ttcdnxut32,33,39(c,d,e)caARTlnhnghpchtlnutinc nghin cu v iu
ch Vit Nam tha mn cc yu cu v tng hp v ha hc. V mt qui trnh tng hp,
m bo tnh n nh, cc bc d thc hin (nhit phng, thi gian va phi,
hachtphbin),hiusutphnngcaovrtcao(85-97%).Vmt ha hc cc hp cht trn va
c bn cao (lin kt 10-C-C) v kh nng tan tt (to mui tan nh nit)
[37].1.4.2.3. Tnh cht ca 5 dn xut
C5dnxututantttrongncvtrongduhnARTnhDimethylsulfoxide (DMSO), c bit
cc dn xut 32, 33 tan tt trong C2H5OH, cn cc dn xut 39c, 39d, 39e th
tan tt trong CH2Cl2 [37]. bn ha hc cao, bn vng hn trong mi trng pH
acid d dy [27].Mt s ch s c bn v c tnh l, ha c bn ca 5 dn xut c nu
chi tit bng 1.1 sau:Bng 1.1. c trng l ha c bn ca 5 dn xut [37].STT
Tn hp cht TLPT (gam/mol) tinh khit (%)1 NT 32 392 992 NT 33 392 993
NT 39c 416 954 NT 39d 416 955 NT 39e 429 95CHNG 2.I TNG V PHNG PHP
NGHIN CU2.1. NGUYN VT LIU, THIT B 2.1.1. Chng k sinh trng st rt
KSTSRP.falciparumkhngchloroquin(K1)ccungcpbiKhoaNghincuiutr st rt,
Vin St rt K sinh trng Cn trng Trung ng v c nui cy lin tc trong
lab.2.1.2. ng vt th nghim 130 con chut nht trng (Mus musculus L.),
chng Swiss, thun chng.Tiuchun:Trngthnh,khemnh,c 2 ging c ci (ng vt
ci phi khng c
mangthai,chasinhsnlnno),trnglng202g,doVinvsinhdchtTrungng cung
cp.2.1.3. Thuc v ha cht nghin cu Bt 5 dn xut ca ART 32, 33, 39c,
39d, 39e do Vin ha hc Vit Nam cung cp:10-[2-hydroxy-3-imidazol)
propyl] deoxoartemisinin (32).10-[2-hydroxy-3-imidazol) propyl]
deoxoartemisinin (33).10-(deoxoartemisinin)-N-((pyridin-3-yl)
methyl) acetamide (39c).10-(deoxoartemisinin)-N-((pyridin-2-yl)
methyl) acetamide (39d).10-(deoxoartemisinin)-N-phenethylacetamide
(39e).Bt ART c tinh khit cao do Vin kim nghim cung
cp.MitrngnuicyRPMI1640(RP)cahngGibco(Hepes,glucoza,gentamicin,
NaHCO3 5%), RPHS (mi trng RPMI, NaHCO3, huyt thanh ngi 10%).Hng cu
v huyt thanh ngi nhm O.Dch treo 40% (4ml cn HC lnh + 6ml RPHS), dch
treo 4% (50l dch treo 40% + 450 l RPHS).Dung mi ha tan: Dimethyl
sulfoxide (DMSO) ca hng Sigma, bt gm arabic
1%.Hachtkhc:Ethanol,methanoltuyti, Giemsa, du soi, m phosphate
pH=6,8; dung dch acid picric 1%, nc ct, dung dch chng ng herparin,
sorbitol 27%, sorbitol 5%.2.1.4. Dng c v thit b dng trong nghin cu
Phin nha 96 ging v trng, bnh nn (Desicator), t sy, ni hp, t m.Bnh
ng ni t lngKnh hin vi, lam knh, pipet Dragendorff, pipet cc loi.Cn
in t, my o pH.Bdngcphathuc:pipetEppendorf,bchyci,ngong,cccm,bnhnn,a
thy tinh.B dng c m chut: bng, cn 70, ko, xylanh 1ml, knh lp,
panh2.2. NI DUNG NGHIN
CUNuicyKSTSRP.falciparumkhngCHQlintctrongphngthnghimnnh chng trc
khi tin hnh thc nghim. Th nghim in vitro ca 5 dn xut 32, 33, 39c,
39d, 39e trn chng P.falciparum khng CHQ theo phng php th 48 gi ca
Nguyn nhPhcvchnrachtchottnhchngSRmnhnhtthctnhcptrn ng vt.Th c tnh
cp ca cht c hot tnh in vitro cao nht trn ng vt thc nghim cc mc liu
dng khc nhau theo ng ung.2.3. PHNG PHP NGHIN CU2.3.1. Th nghim in
vitro ca 5 dn xut 32, 33, 39c, 39d, 39e trn chng P.falciparumkhng
CHQ nui cy 2.3.1.1. Nui cy KSTSR P. falciparum lin tc trong phng th
nghim [5]ChngP.falciparumkhngCHQ(K1)cnuilintctronglabtheophngphp
bnhnncaTragervJensen,thchintrnnguyntcvtrngtrongthuthpmuvnui cy.Cc
bc tin hnhBc 1: Chun
bChunbmitrngnuicyy(RPHS):TmitrngRPMI1640(RP)thm NaHCO3 5% (42l/ml),
huyt thanh ngi nng 10%. Bo qun 4C trong vng 1
tun.Chunbhuytthanh:Mukhngchngngchovochaihoclthytinh,lnm nghing nhit
phng 1 2 gi sau t lnh 4C qua m, dng pipet ht nh ni trn chia vo l nh
theo yu cu cng vic nu c ln HC th li tm, bt hot huyt thanh 56C trong
30 pht, ngui bo qun t lnh trong 30 ngy.Chun b HC lnh: Mu tnh mch c
cha cht chng ng sau khi li tm 2 ln, ht b phn huyt thanh trn, thm mi
trng RP theo t l 5:1, li tm 2000 vng/ 5 pht, ht b phn
ni,ra2lnvimitrngRPvraln3vimitrngyhuytthanh10% (RPHS).Trncn hng cu
vi mi trng RPHS c dch treo 40% (4:6). Chia nh vo l, bo qun 4C trong
30 ngy.ChunbHCnhim:Dngpanhkplymucha500lmucchngP.falciparumK1
cnnuicytrongbnhnitlng,tanngbngtmtrong5pht.Chomuvotubeli
tm,thmnglngsorbitol27%(500l),li tm 2000 vng/5 pht. Ht phn trong
trn, trnuvi1000lsorbitol5%,litm2000vng/5pht(2ln).Hthtphnnitrn,
thm5mlRPHS,litm2000vng/5pht(lm3ln).Htbphnni,thmmitrng RPHS vi 20%
huyt thanh (2ml mi trng + 200l HC lnh) trn u, cho ra a nui.Bc 2:
Nui chngCho a nui vo bnh nui, t nn i khi nn gn tt, kha vi ca bnh
nui to nng kh: 5% CO2, 5% O2 v 90% N2.t bnh nui vo t m 370,5C.Bc 3:
Thay mi trng v ly lam mu theo di hng ngy Pha mi trng RPHS v t vo t
m t nht 30 pht trc khi thay mi trng hng ngy. Ly bnh nui ra khi t m,
m kha, ly ging hoc a ln bn
inox.NhnhngnghinganuitrnhxotrnHCvmitrng,dngpipetPasteurvtrng ht b
phn mi trng pha trn (trnh ht HC).Dng pipet v trng ly khong 2l lm
tiu bn lam git n.Sau , cho mi trng mi vo, lc nh trn u, cho vo bnh
nui, t nn, kha vi v t vo t m.Khi lam kh, c nh bng cn methanol tuyt
i, nhum Giemsa vi dung dch pH=7,2. lam kh t nhin hoc dng my sy.Bc
4: Soi lam v tnh mt k nhim sinh trng Soi lam di knh hin vi vt knh
du 100.m HC nhim KST th v tnh theo 10000 HC (khong 30 vi trng).T l
HC b nhim KST = x100 (%)Bc 5: Cy chuyn
ChunbHClnh:TnhtonslngHClnhcndngthay,phalongHClnhvi mi trng RPHS
thnh dch treo 4% (pha long dch treo 40% vi mi trng RPHS 10 ln )H mt
KST nhim xung cn 0,5% bng cch trn HC v a vo nui cy, 3 ngy cy chuyn
1 ln.2.3.1.2. Th nghim in vitro theo phng php th thuc 48 gi [5]Bc
1:Chun b thuc th: Ha tan 5 dn xut 32, 33, 39c, 39d, 39e vo trong
dung mi DMSO c cc dung dch gc cng nng 10-2 mol/L. Sau , t dung dch
gc pha long vo trong
mitrngRPHScccdynngccthucthlnltl3,10,30,100,300, 1000 nmol/L. Tin
hnh tng t vi ART thu c cc mu c nng tng
ng.Chunbmuth:NuicyKSTSRP.falciparumkhngCHQ(K1)lintctrongmi
trngRPHStheophngphpTragerJensentrnhbymc2.2.1.1.MtKST trc khi th 2
ngy phi t n 3 - 5% HC nhim. Bc 2:Tin hnh th trn phin nha 96 ging,
thc nghim c tin hnh 2 ln.Mi ging trn phin nh 100l mi trng RPHS v
100l thuc cc nng khc nhau (mi nng th 2 ging). Sau nh thm 10l dch
treo KST (40%) vo cc ging to thnh dch treo 4% vi mt 0,3 0,5%. Lc nh
phin nha trn KST v mi trng. t cc phin nha vo bnh nn, t nn to nng kh
thch hp: 5% CO2, 5% O2 v 90% N2 v nui trong t m 37C0,5; sau 24h
thay kh bnh nn 1 ln.Sau 48 gi, ly mu nui ra, lm lam git mng mi
ging, nhum Giemsa, soi knh hin vi v m s lng KST trn tng lam git mng
theo 10.000 HC cc ging chng v cc
gingcnngthuckhcnhau,tnhmtHCnhim.Txcnh nng c ch 50% (IC50) v nng c
ch ti thiu KST pht trin (MIC).Ch tiu nh
gi:Xcnhnngcch50%KSTphttrin(InhibitoryConcentration50%IC50) da trn
chng trnh ANOVA ci sn.T l c ch KST = Xc nh nng c ch ti thiu
(Minimum Inhibitory Concentration MIC) 2.3.2. Th nghim c tnh cp ca
dn xut c hot lc in vitro mnh nht [1], [5], [25]
Thnghimctnhcpngungcachutnhttrngctinhnhtheophng php c
in.ChunbVTN:Chutsaukhimuavcnuigitrongiukinmitrngth nghim, yn tnh,
thong kh, nhit t 20 30C, m ph hp 50 60%, hn ch ting n, chung nui gi
phi c kch thc ph hp, thc n v nc ung, thong, sch s. Chut c nui n nh
trong 2 3 ngy trc khi tin hnh th nghim v trc khi dng thuc trong tnh
trng nhn i qua m (12 15 gi), sau khi dng thuc khng cho n 1 2 gi.
Khi lm thc nghim nh du chut bng dung dch acid picric 1%, chia ngu
nhin 130 con vo cc l, mi l 10 con.Chunb mu th: Bt cht 32 nghin nh
bng ci ri phn tn vo dung mi gm Arabic 1% c pha sn. Dung dch thu c
dng hn dch.ng dng thuc: Th theo ng ung, a thng thuc vo d dy bng kim
u t.S ln v th tch dng thuc: Dng 1 liu duy nht trong ngy, th tch 01
ml/1 gam chut.Tin hnh:Chut nhn i qua m (16h), cn trng
lng.Tinhnhviccliuthnghim:800,1000,1300,1500,2000,2500,3000,3500,
4000, 4500, 4800, 5000, 5200 mg/kg.Cho chut ung thuc, sau khi ung 2
gi cho n ung li bnh thng.Theo di chut trong 1 tun sau khi dng thuc.
Ch tiu quan st gm :Biuhinctnhcpcbitngaysaukhidngthuc,nhngbtthngv
hnhvi,vnngnhili,cng,cogit;biuhincaccbphn,gic quan nh mt, ui,
lng.Theoditnhtrngsckhe:khnngtiuththcn,ncung,tnhtrng phn.Trng lng
chut: trc v sau khi kt thc th
nghim.Slngchutbcht(nuc)ngvimcliuth,duhiusuykit,nhim c nng, hp hi ko
di, khng c kh nng sng st th tnh nh trng hp con vt b
cht.Mquanstithvittcchutbchttrong thi gian theo di v c nhng con chut
cn sng st sau thi gian theo di xem cc c quan: tim, gan, thn, phi,
bng quang v rut xem c g bt thng khng.Ch tiu nh gi:Liu LD0 mg/kg:
liu cao nht khng gy cht chut.Liu cht trung bnh (LD50) mg/kg: liu gy
cht 50% s sc vt th nghim c
tnhtheochngtrnhProbit(Probitanalysisprogram)caMichelRaymond, 1987
(Php).Liu cht tuyt i (LD100) mg/kg: liu gy cht 100% s sc vt th
nghim.Du hiu ng c, biu hin bt thng.Quanstithcccquan: tim, gan, thn,
phi, bng quang, rut chut cht do thuc.CHNG 3.THC NGHIM, KT QU, BN
LUN3.1. KT QU NGHIN CU 3.1.1. Th nghim hiu lc in vitro ca 5 dn xut
32, 33, 39c, 39d, 39e Nui cy P. falciparum khng CHQ (K1) trong lab
lun c duy tr lin tc v trc khi ththuc2ngyphit mt KST 3 5%. Di y l kt
qu duy tr chng K1 trong 10 ngy trc khi th thuc.Bng 3.1. Mt KST chng
P. falciparum khng CHQ trong 10 ngy trc khi th thucNgyTng s HC mS
HC nhimMt KST (%)Lu 1 10000 240 2,42 10000 750 7,5Cy chuyn xung
0,5%3 10000 50 0,54 10000 290 2,95 10000 580 5,8Cy chuyn xung 0,5%6
10000 90 0,97 10000 370 3,78 10000 620 6,2Cy chuyn xung 0,5%9 10000
60 0,610 10000 340 3,4Nhn xt:Qua bng 3.1 trong 10 ngy trc tin hnh
thc nghim theo di qui trnh nui chng K1 thy chng c nui duy tr ng qui
trnh, m bo mt KST n nh tin hnh a vo thviccthuctheoyucu.KtquvmtKST 2
ngy trc th thuc tha mn mc tiu t ra em i th nghim vi thuc ART v dn
xut.Tip theo tin hnh th hot tnh ca thuc trn KSTP. falciparum khng
CHQ (K1) nui cytheophngphpththuc 48 gi ca Nguyn nh Phc vi dy nng 3
x 10-9, 10 x 10-9, 30 x 10-9, 100 x 10-9, 300 x 10-9, 1000 x 10-9
mol/L. Tin hnh nui cy KST vi 5 dn xut v ART, sau 48 gi tin hnh lm
lam git mng xcnhpngcachngviccthucthbngcchxcnhtlcchKSTcatng thuc th
mi nng nghin cu. Kt qu th hin cc bng sau:Bng 3.2. S p ng ca chng P.
falciparum K1 trn in vitro vi cht 32GingCM(nmol/L)T l c ch KST
(%)SDCM1 2 3 4 32 ARTCM(nmol/L)32 ART 32 ART 32 ART 32 ART3 37 30
34 25 32 18 35 32 34,5 1,0 26,23,110 46 49 43 43 47 38 42 47 44,5
1,2 41,8 2,730 68 62 61 70 62 68 65 62 641,6 65,62,1100 79 84 79 91
81 80 89 81 822,4 842,5300 98 95 95 98 92 87 96 95 951,3
93,82,31000 100 100 100 100 96 96 100 100 991,0 98,71,3Biu 3.1. T l
c ch chng P. falciparum K1 ca cht 32Nhn xt:Bng 3.2 v biu 3.1 cho
thy: Hotcht32nngthpnht(3nmol/L)ctcdngcchchngK1vitl 34,5%, khi nng
cng cao th t l c ch KST ca cht 32 cng tng. nng 300 nmol/L th t l c
ch KST t 95%, nhng khi tng ln nng cao nht l 1000 nmol/L th vn ch
t99% cha t mc 100%. So snh vi t l c ch chng K1 ca ART th t l c ch
KST cacht322nngthp(3v10nmol/L)caohnART,cnccnngcnlith khng c s khc
bit nhiu.Bng 3.3. S p ng ca chng P. falciparum K1 trn in vitro vi
cht 33GingCM (nmol/L)T l c ch KST (%)SDCM 1 2 3 4(nmol/L) 33 ART 33
ART 33 ART 33 ART 33 ART3 21 30 29 25 27 18 24 32 25,21,7 26,23,110
38 49 38 43 37 38 39 47 380,4 41,8 2,730 62 62 62 70 65 68 64 62
630,8 65,62,1100 79 84 76 91 80 80 81 81 791,1 842,5300 95 95 95 98
97 87 98 95 96,51,0 93,82,31000 100 100 98 100 100 96 100 100
99,50,5 98,71,3Nhn xt:Qua bng 3.3 nhn thy cht 33 nng thp (3 v 10
nmol/L) t l c ch chng K1 ch tdi50%(25,2% v38%)
vtngdnlnnngcaohn.Tuynhin,khisosnhvi ART th t l c ch chng K1 ca cht
33 cc nng hu ht u thp hn t l c ch KST ca ART, cn 2 nng cao nht (300
v 1000 nmol/L) th t l c ch KST li cao hn.Bng 3.4. S p ng ca chng P.
falciparum K1 trn in vitro vi cht 39cGingCM(nmol/L)T l c ch KST (%)
SD12 3 4CM(nmol/L)39c ART 39c ART 39c ART 39c ART 39c ART3 0 30 0
25 0 18 0 32 0 26,23,110 20 49 15 43 12 38 17 47 161,7 41,8 2,730
40 62 38 70 41 68 39 62 39,50,6 65,62,1100 50 84 59 91 52 80 51 81
532,0 842,5300 98 95 97 98 99 87 100 95 980,6 93,82,31000 100 100
100 100 100 96 100 100 100 98,71,3Nhn xt:Qua bng 3.4 cho thy cht
39c nng 3 nmol/L khng c kh nng c ch chng K1
vchbtuctcdngcchtnng10nmol/L(16%), sauthtngmnhvn nng1000thtl c ch t
100%.So snh vi ART th 4 mc nng u 3; 10; 30;
100nmol/LtlcchchngK1kmhnhn,nhngnnng300v1000nmol/Lth cng cao hn.Bng
3.5. S p ng ca chng P. falciparum K1 trn in vitro vi cht 39dGingCM
(nmol/L)T l c ch KST (%)SDCM 1 2 3 4CM (nmol/L)39d ART 39d ART 39d
ART 39d ART 39d ART3 0 30 0 25 0 18 0 32 0 26,23,110 6 49 0 43 5 38
0 47 2,751,6 41,8 2,730 39 62 28 70 28 68 40 62 33,73,3 65,62,1100
87 84 88 91 90 80 96 81 902,0 842,5300 98 95 98 98 99 87 100 95
990,4 93,82,31000 100 100 100 100 100 96 100 100 100 98,71,3Nhn xt:
Qua bng 3.5 cho thy cht 39d nng 3 nmol/L cng khng c kh nng c ch
chng K1. 2 nng 10 v 30 nmol/L, t l c ch chng K1 di 50%, sau bt u
tng mnh 3 nng cui (100; 300; 1000 nmol/L) t 90%; 99%; 100%. So snh
vi t l c ch KST ca ART th hon ton khc bit, 2 nng u 3 nmol/L, 10
nmol/L cht 39d thp hn ART20 ln, nhng n 3 nng cui th li c kh nng c
ch KST cao hn.GingCM (nmol/L)T l c ch KST (%)SD1 2 3 4CM
(nmol/L)39e ART 39e ART 39e ART 39e ART 39e ART3 21 30 24 25 21 18
25 32 22,71 26,23,110 42 49 43 43 39 38 42 47 41,50,9 41,8 2,730 59
62 63 70 59 68 63 62 611,1 65,62,1100 79 84 85 91 86 80 87 81
84,21,8 842,5300 87 95 97 98 94 87 99 95 94,32,6 93,82,31000 98 100
100 100 99 96 100 100 99,20,5 98,71,3Bng 3.6. S p ng ca chng P.
falciparum K1 trn in vitro vi cht
39eNhnxt:Quabng3.6tathynngcngcao,tlcchchngK1cacht39ecng cao. Tng t
cht 32, 2 nng u t l c ch chng K1 u di 50% v tng dn cc nng cao hn.
So snh vi ART th t l c ch chng K1 ca cht 39e hu ht cc nng u khng c
s khc bit nhiu, ch ring nng 3 v 30 nmol/L l c kh nng c ch KST thp
hn ART.TtlcchKSTcaccchtART,32,33,39c,39d,39ecxlsliubng chng trnh
Probit v ANOVA xc nh nng c ch 50% KST pht trin (IC50) v nng c ch ti
thiu (MIC), kt qu thu c th hin cc bng sau:Bng 3.7. Nng c ch 50% KST
pht trin ca 5 dn xut chng K1Mu th IC50 (nmol/L) PART (1) 16,31,832
(2) 14,53,6 P2,1 > 0,0533 (3) 33,81,8 P3,1 < 0,0539c (4)
91,22,9 P4,1 < 0,0539d (5) 65,14,8 P5,1 < 0,0539e (6) 27,80,7
P6,1 < 0,05 Biu 3.2. Gi tr IC50 ca ART v 5 dn xut trn chng P.
falciparum K1Nhn xt: Qua bng 3.7 v biu 3.2 cho thy:Trong 5 dn xut
32, 33, 39c, 39d, 39e em i th nghim, dn xut 39c c nng c ch 50% KST
pht trin cao nht (91,22,9 nmol/L), cht 32 c IC50 thp nht (14,53,6
nmol/L). So snh vi IC50 ca ART th IC50 ca dn xut 39c cao gp5 ln (s
khc bit c ngha thng k vi P < 0,05); cc dn xut 33, 39d, 39e c
IC50 nh hn IC50 ca cht 39c nhng vn
caohnIC50caART(skhcbitcnghathngkviP 0,05).Nh vy trong 5 dn xut th
cht 32 l cht c tc dng c ch P. falciparum K1 tt nht
vtngtART.Cnccdnxut33,39c,39d,39euctcdngcchchngP. falciparum K1 thp
hn dn xut 32 v ART.Bng 3.8. Nng c ch ti thiu ca 5 dn xut trn chng
P. falciparum K1Mu th MIC ( nmol/L)ART 10032 10033 10039c 30039d
30039e 100Biu 3.3. Gi tr MIC ca ART v 5 dn xut trn chng P.
falciparum K1Nhn xt:Qua bng 3.8 v biu 3.3 cho thy:Nng c ch ti thiu
(MIC) ca 5 dn xut 32, 33, 39c, 39d, 39e phn lm 2 nhm v c chnh lch r
rt . Nhm 1 gm cht 32, 33, 39e vi MIC = 100 nmol/L v nhm 2 gm cht
39c, 39d vi MIC = 300 nmol/L, trong nhm 2 c MIC gp 3 ln MIC nhm 1.
Nh vy, phinnggp3nhm1,nhm2mickhnngcchtithiuchngK1,iuny chng t hot
tnh ca nhm 2 km hn nhm 1 nhiu.So snh vi MIC ca ART th nng c ch ti
thiu (MIC) ca nhm 1 bng MIC ca ART (100nmol/L), cn MIC ca nhm 2 cao
gp 3 ln ART (300 nmol/L).Quacc kt qu IC50 v MIC ca 5 dn xut th v
ART, nhn thy rng cht 32 c hot lc in vitro trn chng P. falciparum K1
mnh nht trong 5 mu th v tng t ART. V vy, chng ti la chn dn xut 32
th nghim c tnh cp trn VTN.3.1.2. Th nghim c tnh cp ca hp cht 32
T130conchutthcnghimcchiangunhinthnh13nhm(10 con/nhm). Sau
,chutcchoungcht32ccnngkhcnhautheophngphpcinchokt qu bng sau:Bng
3.9. Kt qu sau 7 ngy theo di chut ung cht 32 nng khc nhauL Liu dng
(mg/kg) n S chut cht1 800 10 0/102 1000 10 0/103 1300 10 0/104 1500
10 0/105 2000 10 0/106 2500 10 0/107 3000 10 0/108 3500 10 0/109
4000 10 0/1010 4500 10 0/1011 4800 10 0/1012 5000 10 0/1013 5200 10
0/10Nhn xt:Sau khi cho chut ung thuc v theo di trong 7 ngy, chut vn
n ung, hot ng bnh
thng,bititbnhthng,lngmt,khngcbiuhinbtthngvkhngcchutt vong vi tt c
nng thuc th t 800 mg/kg n 5200 mg/kg.Ton b chut sau khi theo di 7
ngy c m quan st i th cc c quan: tim, phi, gan, thn, rut, bng quang.
Kt qu c th hin bng 3.10.Bng 3.10. Kt qu quan st cn nng v i th cc c
quan khi m ton b c 13 l chut sau 7 ngy theo diCh tiu quan st Biu
hin T lCn nng Khng thay i ng k 100%Tim Bnh thng 100%Gan Bnh thng
100%Thn Bnh thng 100%Phi Bnh thng 100%Bng quang Bnh thng 100%Rut 2
con b xut huyt nim mc nh 1,54%Nhn
xt:Saukhiquanstcnnngvithcccquanchut:tim,phi,gan,thn,rut,bng quang
th chng ti nhn thy cn nng v cc c quan u bnh thng tr 2 con trong l
13 b xut huyt nim mc nh rut chim 1,54% s chut th nghim.3.2. BN
LUN3.2.1. Kt qu th hot lc in vitro ca 5 dn xut 32, 33, 39c, 39d,
39eTinhnhnghincuthnghimnhgitcdngca5dnxut10-[(2-hidroxy-3-imidazol)propyl]deoxoartemisinin(32),10-[(2hidroxy-3-imidazol)propyl]
deoxoartemisinin(33),10-(deoxoartemisinin)-N-((pyridin-3-yl)methyl)acetamide(39c),10-(deoxoartemisinin)-N-((pyridin-2-yl)methyl)acetamide(39d),10-(deoxoartemisinin)-N-phenethylacetamide(39e)iviKSTP.falciparumkhngCHQnuicytrninvitrothng
qua vic xc nh cc ch s nh gi l nng c ch 50% KST pht trin (IC50) v
nng c ch ti thiu
(MIC).Tktqunghincuchothyc5dnxutuctcdnginvitrovichngP.
falciparumkhngCHQ(K1).Mcdkhnngcchcaccchtkhcnhau,nhnguc
cimchunglnngcngcaothkhnngcchKSTcngcaov2nngcao nht 300; 1000 nmol/L
u c kh nng c ch KST cao hn ART. Qua t l c ch KST ta xc
nhcIC50nhgirhnvhottnhinvitroca5dnxutvsosnhviART.
Trong5dnxutthcht39ccIC50caonht(91,22,9nmol/L) vdnxut32cIC50thp
nht(14,53,6nmol/L)chngtcht32chotlcinvitromnhnht,cncht39cchot lc in
vitro km nht trn chng P. falciparum K1. Bn cnh , IC50 ca cc dnxut
33, 39c, 39d, 39e cao hn ART nn 4 dn xut ni trn c hot tnh in vitro
trn chng P. falciparum K1 thphnART.RingchcIC50cacht32thp hn IC50 ca
ART: 14,53,6 nmol/L so vi 16,31,8 nmol/L nhng s khc bit khng c ngha
thng k vi P > 0,05; iu ny chng t cht 32 c hot lc in vitro i vi
chng P. falciparum K1 tng t hot lc ca ART. Ngoi ra, theo cc nghin
cu v IC50 ca ART trc y thu c trong iu kin nghin cu ti lab l: 14,5
nmol/l (Nguyn Vit Hng, 1995) v 15 nmol/l (Nguyn Mai Hng, 2001) [9]
th kt qunghincucachngticaohnnhngkhngnhiu.Bncnh,IC50cadnxut32 thp hn
PQ (IC50 = 46,2 nmol/l) trong cng iu kin th nghim ti Vin st rt nn
hot tnh in vitro ca cht 32 cao hn PQ trn chng K1 [18]. ng thi, so
snh vi thc nghim ca tc gi Nguyn Th Minh Thu v cng s v hot lcin
vitro ca 5 dn xut 32, 33, 39c, 39d, 39e
vichngK1trongcngiukinthcnghim[19]thktqucachngtivhotlcin vitro ca 5
dn xut khng c s thay i nhiu sau 4 nm v iu ny cng khng nh tc dng ca
5 dn xut trn i vi chng K1, trong ni bt l dn xut
32.QuaxcnhnngcchtithiuMICca5dnxutth3dn xut c nng c
chtithiuvichngKSTP.falciparumkhngCHQnng100(nmol/L)vbngMIC ca ART l
32, 33, 39e; cn 2 dn xut 39c, 39d c MIC l 300 nmol/L cao hn MIC ca
ART v dn xut cn li, s khc bit c ngha thng k vi P < 0,05. iu ny
chng t rng 2 dn
xut39cv39dctcdngvichngP.falciparumkhngCHQ(K1)kmhnccdnxut cn li v
ART. Qua ch s IC50 v MIC thu c kt qu nghin cu, chng ta nhn thy: C 5
dn xut
uchottnhinvitrotrnchngP.falciparumkhngCHQ(K1),trong5dnxutthdn xut
39c, 39d c gi tr IC50 v MIC cao hn cc cht khc th hin hot tnhin
vitro km hn ccdnxutcnlivkmhnARTnhiuln.Ngoira,dnxut32lchtchottnhin
vitrovichngK1caonhtvtngtARTvchothycht32lchtcnhiutrinvng nht trong 5
dn xut c iu ch theo phng php mi v ln u tin c tng hp thnh cng Vit
Nam.3.2.2. Kt qu nghin cu c tnh cp dn xut 32 - cht c hiu lc in
vitro mnh nht Theoktqunghincuinvitro,chngtilachncht32th nghim c tnh
cp trn chut nht trng.y l mt th nghim khng th thiu nh gi an ton ca
thuc nhm phn loi mc c ca thuc . Trong cc php th nh gi c ca thuc th
phpthctnhcplphpthutinc thc hin. Trong nghin cu c tnh cp ca cht 32
trn chut nht trng chng ti dng ng ung, y l ng d kin s dng trn ngi v
s dng phng php th c in. y l phng php c chnh xc cao, ph hp vi yu cu
d liu, c bit s liu c chy theo chng trnh Probit ci sn nn kt qu rt ng
tin cy trnh sai s do tnh ton. Vi m hnh th ca phng php th c in chng
ti thnghimvidyliumcliuthpnhtl 800 mg/kg v liu cao nht l 5200 mg/kg.
Sau khi theo di 7 ngy, quan st cc du hiu ng c chng ti nhn thy rng
tt c cc liu th nghim khng c chut no t vong, m quan st i th cc c
quan (tim, gan, phi, rut, thn, bng quang) v cn nng hu ht chut khng
c biu hin bt thng, ch c 2 con chut
trongl13bxuthuytnimmcnhrutchimtl1,54%.Dothigiannghincuc
hnnnchngtichdngliquanstith,chaciukinnghincusuhnv2 trng hp b xut
huyt nim mc rut, nn vn cha c kt lun chnh xc v nguyn nhn gy xut huyt
nim mc rut nh ca 2 con chut trn, v vy cha th kt lun chnh xc l trng
hp ng c cp. Tuy nhin chut b xut huyt nh v chim t l rt thp (1,54%),
nn c th sbnhgidnxut32gnnhkhngcctnhcpmcliu5200mg/kg theo phn
loiGHS(GloballyHarmonisedClassificationSystem for Chemical
Substances and Mixtures). Tuy nhin y ch mi l nghin cu bc u trn ng
vt nh, nn cn phi tip tc tin hnh
thnghimlmsngtrnngvtlnhnvngiviccphngphpthctnhkhc trc khi a vo s dng
trong iu tr bnh SR. Mc d vy, kt qu thu c t nghin cu ca chng ti cng
rt kh quan, to c s cho cc nghin cu sau ny v hp cht 32.T cc kt qu
nghin cu in vitro v th c tnh cp cho thy rng, hp cht 32 c hot
tnhinvitroivichngP.falciparumkhngCHQ(K1)caonhttrong5dnxutthv tng ng
vi ART, ng thi hu nh khng c c tnh cp. Bn cnh , dn xut 32 vi cu trc
c cha lin kt 10-C-C, nit nn c kh nng tan tt trong nc v trong du, bn
vng ha hc hn ART nn ha hn l 1 hot cht c th a vo iu tr SR khng thuc
trong tng lai.Mcdychlnhngktqubanunhngphn no khng nh c thnh cng
bcucaccnh khoa hc Vit Nam trn con ng tm ra cc hp cht mi c hiu qu iu
tr bnh SR ti Vit Nam.KT LUN V KIN NGHKT LUNQua nghin cu hiu lc in
vitro ca 5 dn xut 32, 33, 39c, 39d, 39e do Vin Ha hc sn xut vi chng
P. falciparum khng CHQ (K1) v c tnh cp ng ung ca dn xut
10-[2-hydroxy-3-imidazol)propyl]deoxoartemisinin(32)trnchutnhttrng,chngtihon
thnh mc tiu ti t ra v thu c kt qu sau:C 5 dn xut 32, 33, 39c, 39d,
39e em th u c hiu lc in vitro vi chng P. falciparumkhng CHQ (K1).
Dn xut 32 l dn xut c hiu lc cao nht vi IC50 = 14,53,6 nmol/L, MIC =
100 nmol/L v tng t hiu lc ca ART.Thctnh cp ca hp cht 32 (cht c hiu
lcin vitro mnh nht) trn 130 chut nht trng bng ng ung vi dy liu dng
t 800 mg/kg n 5200 mg/kg, khng c chut no cht, theo di cc du hiu ng
c v quan st i th cc c quan nh tim, gan, thn, phi, rut, bng quang v
cn nng hu ht khng thy biu hin bt thng.KIN NGHT kt qu nghin cu trn
chng ti c mt s kin ngh sau:M rng nghin cu tc dng ca 5 dn xut 32,
33, 39c, 39d, 39e c bit l hp cht 32 i vi ng vt th nghim tm ra hot
lc, tc dng ca cc dn xut trn in vivo, t
tmradnxutttnhtcthavoiutrvtmraliuiutrphhpvi chng KSTSR khng thuc.Tip
tc nghin cu thm v c tnh cp, bn cp v c tnh trng din ca hp cht 32 nhm
mc ch hon thin qu trnh nghin cu tin lm sng to iu kin cho giai on
nghin cu lm sng trn ngi tip theo.TI LIU THAM KHOTi liu ting Vit
Trung m (1996), Phng php xc nh c tnh cp ca thuc, Nh xut bn y
hc.Hong H v cng s (2010), Hiu qu ca bin php truyn thng phng chng st
rt vi ngi Vn Kiu ti x Thanh, huyn Hng Ha nm 2006, Tp ch phng chng
cc bnh st rt v cc bnh k sinh trng, (s 4), tr. 3.Trn c Hu (2007), Ha
dc tp 2, Nh xut bn y hc, tr.
213.LXunHng,NguynMnhHng(2010),Bnhstrtvchinlcphngchng,Nh xut bn y
hc.NguynMnhHng(2011),Cmnangkthutphngchngbnhstrt,Nhxut bn y
hc.NguynMnhHngvcngs(2011),HiuquphngchngstrttiVitNamgiai on 2006
2010, Cng trnh nghin cu khoa hc bo co ti hi ngh k sinh trng ln th
38 tp 1, Nh xut bn y hc, tr. 9
-14.NguynXunHng(2012),iutrakhngChloroquincaP.falciparumbngkthut PCR
ti hai tnh Qung Tr v knng nm 2008 2009, Tp ch phng chng cc bnh st
rt v cc bnh k sinh trng, (s 1), tr. 39 43.L Th Thu Hng (2009), K
sinh trng, Nh xut bn gio dc Vit Nam, tr. 108 135.NguynMaiHngvcngs
(2001), nh gi s nhy cmin vitro v hiu lc iu tr
cathucstrttrnbnhnhnstrtP.falciparumtitnhBnhPhc,Kyu CTNCKH Vin St rt
K sinh trng Cn trng Trung ng(1996-2000), Nh xut bn y hc, tr. 309
315.JanA.Rozendaal(2000),Phngchngvttruynbnh,Nhxutbnyhc,tr.20-25.
Ngi dch Phm Huy Tin v cng
s.NguynVnKim(1993),DchthcstrtkhngthucVitNam,Thngtinphng chng bnh st
rt v cc bnh k sinh trng, (s 1), tr. 19
21.LBLcvcngs(2006),Nghincuksinhtrngstrtkhngthucvnhgi hiu lc mt s
phc chng khng ti mt s khu vc min Trung Ty Nguyn, Cng trnh nghin cu
khoa hc bo co ti hi ngh khoa hc ton quc chuyn ngnh st rtk sinh trng
cn trng giai on 2001 2005 tp 1, Nh xut bn y hc, tr. 180
187.HuNgh,NguynVnHn(2006),Tnghphaidnxut10arylamino Artemisinin, Tp
ch dc hc, (s 363), tr. 8 -
10.PaulN.Newtonvcngs(2008),Hptcnghincudchthcvtiphmbunbn thuc
Artesunate gi ti cc nc ng Nam , Tp ch phng chng cc bnh st rt v cc
bnh k sinh trng, (s 5), tr. 20 22. Bn dch ting Vit ca Trnh Ngc Hi.V
Th Phan (1996),Dch t hc bnh st rt v phng chng st rt Vit Nam, Nh xut
bn y hc, tr. 17 147.Bi Quang Phc (2008), Nghin cu c cu k sinh trng
st rt v Plasmodium falciparum khng thuc bng k thut polymerase chain
reaction ti vng st rt lu hnh nng, Lun n tin s y hc - Hc vin Qun y,
tr.8 - 39.NguynQuangThiu,NguynMnhHng(2012),Mtscimdchtstrtti
huynHngHatnhQungTrgiaion20062010,Tp ch phng chng cc bnh st rt v cc
bnh k sinh trng, (s 1), tr.
3.NguynThMinhThuvcngs(2011),Hiulcvctnhtrnthcnghimca
DihydroartemisininvPiperaquindoVitNamsnxutviksinhtrngstrt,Cng trnh
nghin cu khoa hc bo co ti hi ngh k sinh trng ln th 38 tp 1, Nh xut
bn y hc, tr. 232
240.NguynThMinhThuvcngs(2011),Nghincuhiulcinvitroca5dnxut
Artemisinin v c tnh cp ca 10-[(2hydroxy-3imidazol) propyl]
deoxoartemisinin, Tp ch dc hc, (s 428)
.NguynThMinhThu(2008),Nghincutcdngtrnksinhtrngstrtvctnh trn thc
nghim ca thuc st rt phi hp Dihydroartemisinin Piperaquin do Vit Nam
sn xut, Lun n tin s dc hc - Vin Dc Liu.Thi Nguyn Hng Thu (2001),
Nghin cu bn tng hp v th tc dng sinh hc ca mt s dn xut mi ca
Artemisinin, Lun n tin s dc hc, Trng i Hc Dc H Ni.Nng Th Tin
(2002), Gim st k sinh trng st rt khng thuc v nh gi hiu lc iu
trcamtsphcdoBYtbanhnh,DnphngchngstrtVitNamEC, Vin St rt K sinh trng
Cn trng Trung ng, tr. 5.Mai Tt T, V Th Trm (2007), Dc l hc tp 2, Nh
xut bn y hc, tr. 193 199.Triu Nguyn Trung v cng s (1997), Hiu lc ca
Artemisinin v Artesunate trongiu tr st rt do P. falciparum ti khu
vc min Trung Ty Nguyn, K yu CTNCKH Vin st rt K sinh trng Cn trng
Trung ng (1991-1996) tp 1, Nh xut bn y hc, tr. 122 131.Vin Kim
nghim (2005), D tho hng dn th c tnh ca thuc, B Y t.Vin SR KST CT
TW, Nghin cu tin lm sng ca hp cht Artemisinin gn Flour, ti cp nh nc
giai on 2002 2004.TrnKhcVvcngs(2006),Phngphpmitnghp10(2-oxoethyl)
deoxoartemisinin, Tp ch dc hc, (s 366), tr. 8 10.Nguyn Xun X v cng
s (2012), Phn tch cc yu t nguy c nhim st rt trong cng ng ngi dn tc
Gia Rai ba thn bin gii: Theo kt qu phn tch khng th khng st rt trong
huyt thanh, Tp ch phng chng cc bnh st rt v cc bnh k sinh trng, (s
2), tr. 15.P.T.Yn,LNguynBnh,ngCmThchv cng s (1997), Hiu lc v hiu qu
ca phc Artemisinin tng liu 60 mg/kg v Artesunate tng liu 12 mg/kg n
thun trong 5 ngy, K yu CTNCKH Vin st rt K sinh trng Cn trng Trung
ng (1991-1996) tp 1, Nh xut bn y hc, tr. 109 113.Ti liu ting
AnhBalintG.A.(2001),Artemisininanditsderivatices,animportantnewclassofantimalarial
agents, Pharmacology and therapeutics, 90,pp. 261 265.Karbwang J.,
Na-Bangchang K. (1995), Clinical pharmacology of artemisinin
compounds, Clinical pharmacology of antimalarials, pp. 263
294.MeshnickS.R.(2002),Artemisinin:mechanismsofaction,resistanceandtoxicity,
International journal for Parasitology, 32, pp. 1655
1660.RobertA.,Benoit-VicalF.,ClaparolsC.,MeunierB.(2005),Theantimalarialdrug
artemisininalkylateshemeininfectedmice,ProceedingsoftheNationalacademyof
science of the United States of America, Vol. 102, No. 83, pp.
13676
13680.RollBackMalariaRBMIssuesBriefOntheglobalmalariaresponseFinr122009
Retrieved from the World Wide Web on 3rd of
June.Quinghaosuantimalariacoordinatingresearchgroup(1979),Antimalariastudieson
quinghaosu, Chinese medical journal, Vol. 92, No. 12, pp. 811
816.TranKhacVu,TruongBichNgan(2007),Synthesisofnovelwatersoluble10deoxoartemisinins,
Journal of Chemistry, Vol. 45(4), pp. 505
508.TruongThiThanhNga,NguyenQuangAn,TruongBichNgan,TranKhacVu(2008),
Synthesis of novel 10 deoxoartemisinin, Vast Proceeding, pp. 201
206.VriesP.J.de,DienT.K.(1996),Clinicalpharmacologyandtherapeuticpotentialof
artemisinin and its derivatives in the treatment of malaria, Drugs,
52(6), pp. 818 835.WilsonJ.F(2003),Advancing the war on
malaria,Annals of Internal medicine, Vol. 138, No. 8, pp. 693
696.World Health Organization (2006), Guidelines for the treatment
of malaria.World Health Organization (2012), Summary and Key
Points, World malaria report 2012.
