B Y T TRNG I HC DC H NI

BI TH HNG NHUNG

NGHIN CU XY DNG CNG THC BO CH VIN NN

ACYCLOVIR KT DNH SINH HC NG TIU HA

KHA LUN TT NGHIP DC S

H NI 2013

B Y T TRNG I HC DC H NI

BI TH HNG NHUNG

NGHIN CU XY DNG CNG THC BO CH VIN NN ACYCLOVIR KT

DNH SINH HC NG TIU HA KHA LUN TT NGHIP DC S

Ngi hng dn: 1. TS. V Th Thu Giang 2. DS. Nguyn Hng Trang

Ni thc hin: B mn Bo ch

H NI - 2013

LI CM N Li u tin, ti xin gi li cm n chn thnh v su sc ti:

TS. V Th Thu Giang Ngi thy ch bo v hng dn ti tn tnh trong sut thi gian

qua, gip ti tng bc nng cao nhn thc v phng php lun hon thnh kha lun tt nghip.

Ti xin chn thnh cm n DS. Nguyn Hng Trang v DSCK II. L Vn Thanh cng cc thy c gio v cc anh ch k thut vin ca b mn Bo ch gip v to iu kin thun li cho ti trong sut thi gian lm thc nghim ti b mn.

Ti cng xin trn trng cm n Ban gim hiu, cc phng ban, cc thy c gio v cn b, nhn vin trng i hc Dc H Ni nhng ngi dy bo v gip ti trong sut 5 nm hc tp di mi trng ny.

Cui cng, ti xin by t lng bit n su sc ti gia nh, bn b nhng ngi lun bn ti, chia s, ng vin v gip ti trong sut thi gian qua. H Ni, ngy 20 thng 5 nm 2013 Sinh vin

Bi Th Hng Nhung

MC LC T VN ................................................................................................................ 1 PHN 1. TNG QUAN ................................................................................................ 3 1.1. S lc v acyclovir ................................................................................................. 3

1.1.1. Cng thc ha hc .......................................................................................... 3 1.1.2. Tnh cht l ha .............................................................................................. 3 1.1.3. Dc ng hc ............................................................................................... 3 1.1.4. Tc dng v c ch ........................................................................................ 4 1.1.5. Ch nh .......................................................................................................... 4 1.1.6. Mt s ch phm acyclovir trn th trng .................................................... 4

1.2. H kim sot gii phng thuc ti d dy (Gastroretentive dosage forms) ......... 5 1.2.1. ng dng ca h kim sot gii phng thuc ti d dy ............................... 5 1.2.2. Cc c ch kim sot gii phng thuc ti d dy ........................................ 6 1.2.3. Mt s h kim sot gii phng thuc ti d dy ........................................... 6

1.2.3.1. H kt dnh sinh hc (Bioadhesive drug delivery systems) .................. 6 a. Khi nim kt dnh sinh hc ............................................................. 6 b. Cc c ch kt dnh sinh hc ............................................................ 7 c. Polyme kt dnh sinh hc .................................................................. 8

1.2.3.2. H thuc ni d dy (Floating drug delivery systems) .................... 11 a. Nguyn tc ...................................................................................... 11 b. Phn loi ......................................................................................... 11

1.2.3.3. H c t trng ln (hay h sa lng) .................................................... 13 1.2.3.4. H trng n hoc thay i hnh dng ............................................... 13 1.2.3.5. H kt hp ni v kt dnh .................................................................. 13

1.3. Mt s nghin cu v h kt dnh sinh hc ng tiu ha cha acyclovir ...... 15 1.3.1. Nghin cu bo ch vin nn ...................................................................... 15 1.3.2. Nghin cu bo ch vi cu .......................................................................... 16 1.3.3. Nghin cu bo ch h nano ....................................................................... 17

PHN 2. I TNG V PHNG PHP NGHIN CU .............................. 18 2.1. Nguyn liu v phng tin nghin cu .............................................................. 18

2.1.1. Nguyn vt liu nghin cu .......................................................................... 18 2.1.2. Phng tin nghin cu ............................................................................... 18 2.1.3. ng vt th nghim ..................................................................................... 19

2.2. Phng php nghin cu ..................................................................................... 19 2.2.1. Xy dng ng chun acyclovir trong mi trng acid clohydric 0,1M.. .. 19 2.2.2. Phng php bo ch vin nn acyclovir kt dnh sinh hc ........................ 19 2.2.3. nh gi mt s ch tiu cht lng ca bt dp vin ................................ 20 2.2.4. nh gi mt s ch tiu cht lng ca vin nn bo ch ......................... 21 2.2.5. Phng php thit k th nghim v x l s liu ........................................ 24

PHN 3. KT QU THC NGHIM V NHN XT ........................................ 25 3.1. Kho st mi tng quan gia nng dung dch v mt quang. ............... 25 3.2. Nghin cu phng php bo ch v cc t dc c bn ca vin nn

acyclovir kt dnh sinh hc .................................................................................. 26 3.2.1. Nghin cu phng php bo ch ............................................................. 26 3.2.2. La chn cc t dc c bn bo ch vin nn acyclovir kt dnh sinh hc .............................................................................................................. 28

3.3. Xy dng cng thc bo ch vin nn acyclovir kt dnh sinh hc .................... 35 3.3.1. Quy hoch thc nghim ............................................................................... 35 3.3.2. nh gi nh hng ca cc bin c lp ti cc bin ph thuc ............. 41 3.3.3. Ti u ha cng thc vin nn acyclovir kt dnh sinh hc ng tiu ha ............................................................................................................................ 45

PHN 4. KT LUN V XUT ........................................................................ 47

DANH MC CC K HIU, CC CH VIT TT K hiu Ni dung ACV Acyclovir

BDDS H thuc kt dnh sinh hc (Bioadhesive drug delivery systems) BP Dc in Anh (British Pharmacopoeia ) CMC Carboxy methyl cellulose

CP Carbopol

CT Cng thc DVN IV Dc in Vit Nam IV EC Ethyl cellulose

FDDS H thuc ni (Floating drug delivery systems) GPDC Gii phng dc cht GRDF H kim sot gii phng thuc ti d dy (Gastroretentive

dosage forms)

HEC Hydroxy ethyl cellulose

HPMC Hydroxy propyl methyl cellulose

HSV Virus Herpes (Herpes simplex virus)

KDSH Kt dnh sinh hc KLPT Khi lng phn t KLTB Khi lng trung bnh MTN Mc trng n Mg stearat Magnesi stearat

NaCMC Natri carboxy methyl cellulose

PVA Polyvinyl alcol

PVP Polyvinyl pyrrolidon

SKD Sinh kh dng TCCS Tiu chun c s TKHH Tinh khit ha hc USP Dc in M (United States Pharmacopeia )

DANH MC CC BNG STT Tn bng Trang

Bng 1 Phn loi polyme kt dnh sinh hc 9 Bng 2 Tnh cht v ng dng ca mt s polyme kt dnh sinh hc 9 Bng 3 Nguyn vt liu nghin cu 18 Bng 4 Mt quang (D) v nng (C) ca dung dch acyclovir 25 Bng 5 Mt quang ca mu cha acyclovir v mu placebo 26

Bng 6 Thnh phn ca vin nn c bo ch vi cc polyme khc nhau

28

Bng 7 Mc trng n ca cc mu vin theo thi gian 28 Bng 8 Kt qu th GPDC v KDSH ca cc mu vin 29 Bng 9 Thnh phn cc cng thc kho st 31

Bng 10 Mc trng n ca cc mu vin kho st 31 Bng 11 Kh nng GPDC v lc KDSH ca cc mu vin kho st 32

Bng 12 Lc KDSH v mc trng n ca cc mu vin c t l natri hydrocarbonat khc nhau

34

Bng 13 Kh nng GPDC v kh nng ni ca cc mu vin thay i t l natri hydrocarbonat 34

Bng 14 Cc bin c lp 36 Bng 15 Cc bin ph thuc 36 Bng 16 Cc cng thc thc nghim 36 Bng 17 Tc chy, ch s nn v hm m ca cc khi bt kp 37 Bng 18 nh gi mt s ch tiu cht lng ca cc mu vin 38 Bng 19 Kh nng GPDC v KDSH ca cc mu vin 39 Bng 20 Kh nng ni v kh nng trng n ca cc mu vin 39 Bng 21 Bng h s tng quan t Y1 n Y8 v KDSH 41

Bng 22 Kh nng GPDC v KDSH ca mu vin bo ch theo cng thc ti u v d on 45

DANH MC CC HNH V, TH STT Tn hnh v, th Trang

Hnh 1 Cht kt dnh lng lan rng trn b mt t bo m mm 7 Hnh 2 S khuch tn ca polyme vo chui glycoprotein mng nhy 8 Hnh 3 Cc c ch ni 12 Hnh 4 S gii phng thuc t h trng n 13 Hnh 5

S cc giai on bo ch vin nn acyclovir kt dnh sinh hc 20

Hnh 6 Thit b nh gi lc kt dnh sinh hc t ch to 22 Hnh 7

th biu din s ph thuc gia mt quang (D) vi nng (C) ca dung dch acyclovir trong mi trng HCl 0,1M 25

Hnh 8 Kh nng trng n ca vin bo ch vi cc polyme khc nhau

29

Hnh 9 th gii phng dc cht ca cc mu vin bo ch vi polyme khc nhau

30

Hnh 10 Kh nng trng n ca cc mu vin theo thi gian 32 Hnh 11 th gii phng dc cht theo thi gian ca cc mu vin 33 Hnh 12

Mt p nh hng ca Carbopol 934P v natri hydrocarbonat ti Y6 42

Hnh 13 Mt p nh hng ca Carbopol 934P v HPMC K100M ti Y8

43

Hnh 14 Mt p nh hng ca Carbopol 934P v natri hydrocarbonat ti lc KDSH 43

Hnh 15 Mt p nh hng ca Carbopol 934P v HPMC K100M ti lc KDSH 44

Hnh 16 th GPDC ca vin bo ch theo cng thc ti u v d on 45

Hnh 17 Hnh nh vin bo ch theo cng thc ti u ni trong dung dch acid clohydric 0,1M 46

1

T VN ng ung l ng a thuc vo c th ph bin nht hin nay. Tuy

nhin, s hp thu thuc qua ng tiu ha kh phc tp. Hp thu thuc thay i dc theo ng tiu ha v chu s tc ng ca nhiu yu t nh enzym, pH, thc nPhn ln cc thuc c hp thu tng i nhanh phn u rut non, n phn cui ng tiu ha th hp thu chm v khng hon ton. V vy, kim sot gii phng thuc vng hp thu ti u trong ng tiu ha l mt trong nhng bin php gip ci thin hp thu v SKD ca thuc.

Acyclovir (ACV) l mt dn cht tng hp ca acid nucleosid guanosin c tc dng chn lc trn t bo nhim virus Herpes, c s dng rng ri trong iu tr mn rp da, vim gic mc, thy u, Herpes sinh dc. Trong mt s trng hp iu tr nhim Herpes ti pht hoc suy gim min dch cn phi s dng ACV ko di ti 6 thng hoc hn. Hin nay, ACV vn l thuc c la chn hng u trong iu tr cc bnh k trnnh c tnh thp i vi t bo ngi v tc dng ph nh hn so vi cc thuc khng virus khc.

Tuy nhin, ACV c tan trong nc ln trong du u hn ch li c tnh thm km, hp thu thuc chm v khng hon ton ch yu on u ng tiu ha, qua khe lin kt t bo. Phn ln thuc b bi tit ra ngoi (50 60%) dng khng c hp thu, SKD ng ung ch t t 10 20% [16]. Do vy nu dng cc dng thuc quy c th phi ung nhiu ln trong ngy (4 6 ln), gy nhiu phin phc cho bnh nhn. Mt trong nhng hng nghin cu ang c pht trin hin nay ci thin SKD ng ung ca ACV l bo ch h kt dnh sinh hc ng tiu ha. H c kh nng ko di thi gian lu thuc ti v tr hp thu ti u trong ng tiu ha nh tng ng k s hp thu thuc, tng cng hiu qu iu tr v gim s ln dng thuc trong ngy nn tng s tun th iu tr ca ngi bnh. Vi nhng u im trn, y l mt trong nhng h thuc c trin vng ci thin c nhng c tnh bt li ca ACV.

2

Hin nay trn th trng trong nc cng nh nc ngoi cha c dng vin nn acyclovir kt dnh sinh hc ng tiu ha. Xut pht t thc t trn nhm pht trin h thuc mi c kh nng ng dng vo thc tin, chng ti thc hin ti: Nghin cu xy dng cng thc bo ch vin nn acyclovir kt dnh sinh hc ng tiu ha vi mc tiu:

Xy dng c cng thc bo ch vin nn acyclovir ni kt dnh sinh hc ng tiu ha gii phng ko di 12 gi.

t c mc tiu ra, kha lun gm mt s ni dung nghin cu chnh nh sau:

1. Kho st nh hng ca mt s polyme v t dc khc n kh nng gii phng dc cht, kt dnh sinh hc, ni v trng n ca vin nn acyclovir.

2. Xy dng cng thc bo ch vin nn acyclovir ni kt dnh sinh hc ng tiu ha.

3

PHN 1. TNG QUAN 1.1. S lc v Acyclovir 1.1.1. Cng thc ha hc

Cng thc phn t: C18H11N5O3 Khi lng phn t: 225,2 Tn khoa hc: 2 amino 9[(2 hydroxy ethoxy) methyl] 1,9 dihydro 6H purin 6 on.

1.1.2. Tnh cht l ha Acyclovir c cc tnh cht l ha sau [2], [10], [33]:

- Dng bt kt tinh mu trng, t tan trong nc, rt t tan trong alcol, tan t do trong cc dung mi dimethyl sulfoxid, tan c trong dung dch kim v acid long.

- tan ca ACV trong nc t 1,2 1,6 mg/ml nhit 22 25C v 2,5 mg/ml 37C v ph thuc vo pH mi trng.

- Rt bn vng, trong mi trng kim n nh hn trong mi trng acid. - Nhit nng chy khong 230C sau b phn hy. - Acyclovir c 2 hng s phn ly: pKa1 = 2,41 0,27 v pKa2 = 9,06 0,88. - Theo h thng phn loi sinh dc hc (BCS), ACV thuc nhm III tan

cao, tnh thm thp [18]. 1.1.3. Dc ng hc

Hp thu: ACV km hp thu qua ng tiu ha, SKD theo ng ung khong 10 20%. Thc n khng lm nh hng n hp thu thuc. ACV c hp thu ch yu theo c ch khuch tn th ng, hp thu chm v khng hon ton. Sau khi ung 1,5 2 gi t Cmax trong mu [1], [5], [33].

Phn b: ACV phn b rng ri trong dch c th v cc c quan nh: no, thn, phi, rut, gan, lch, c, t cung, nim mc, dch m o, nc mt, thy tinh

4

dch, dch no ty. Nng trong dch no ty t ti 50% nng huyt tng. Lin kt vi protein huyt tng thp (9 33%). Thuc qua c nhau thai v phn b trong sa m vi nng gp 3 ln trong huyt thanh m [1], [2], [5], [33].

Chuyn ha v thi tr: Phn ln thuc c o thi qua thn di dng khng bin i. 9

carboxy methoxy methyl guanin l cht chuyn ha ng k ca ACV chim khong 14% tng lng thuc thy trong nc tiu [33].

Thi gian bn thi: ngi ln l 2 3h, tr s sinh l 4h, cn bnh nhn suy thn mn c th ln n 19,5h [1], [5]. 1.1.4. Tc dng v c ch

ACV l mt cht tng t nucleosid c tc dng chn lc trn t bo nhim virus Herpes. Tc dng ca ACV mnh nht trn virus Herpes simplex typ 1v km hn virus Herpes simplex typ 2, virus Varicella zoster v tc dng yu nht trn cytomegalovirus [5].

c tc dng ACV phi c phosphoryl ha thnh dng c hot tnh l acyclovir triphosphat nh cc enzym ca t bo. Acyclovir triphosphat c thu nhn vo DNA virus v tc ng nh l nhnh cui cng v trong cu to ca n khng c nhm 3 hydroxy do vy lm gin on qu trnh tng hp DNA ca virus. ACV c ch s nhn ln ca virus m khng nh hng g n chuyn ha ca t bo bnh thng [5], [33]. 1.1.5. Ch nh

Acyclovir c ch nh iu tr trong cc trng hp sau [5], [33]: - iu tr khi u v d phng nhim HSV typ 1 v typ 2 da, nim mc,

thn kinh v sinh dc, vim no do HSV. - iu tr nhim Herpes zoster cp tnh mt, phi, thn kinh. - iu tr khi u v ti pht Herpes sinh dc. - D phng v iu tr nhim virus ngi suy gim min dch, cy ghp

c quan, bnh thy u.

5

1.1.6. Mt s ch phm acyclovir trn th trng - Vin nn: Apo Acyclovir, Cyclovir, Herpevir, Herpex, Medovir, Vacrax

200mg, Acyclovir Stada, Zovirax 200mg, 400mg, 800mg.

- L bt pha tim: Zovirax 200mg/5ml. - Thuc m dng ngoi Zovirax 5%, thuc m tra mt Zovirax 3% w/w. - Dch truyn: Zovirax tim tnh mch. - Kem bi ngoi da: Acyclovir Stada 5%.

- Vin phn tn: Zovirax 200mg, 400mg, 800mg.

- Vin tc dng ko di: Genvir 600mg. 1.2. H kim sot gii phng thuc ti d dy (Gastroretentive Dosage Forms - GRDF)

1.2.1.ng dng ca h kim sot gii phng thuc ti d dy GRDF l dng bo ch c thit k nhm ko di thi gian lu ca thuc ti

d dy do lm tng s hp thu thuc phn trn ca ng tiu ha nhm nng cao SKD ca thuc. Nh , GRDF gip ko di khong cch gia cc ln dng thuc v tng cng s tun th iu tr ca bnh nhn. Dng bo ch ny c bit c hiu qu vi cc thuc khng tan hoc t tan v vi cc thuc loi ny, khi ko di thi gian vn chuyn thuc trong ng tiu ha s gip gia tng ng k s hp thu thuc [24].

Ngoi ra, GRDF cn rt hu ch trong cc h phn phi thuc nh liposom, ht nano, vi cu, v.v.. So vi cc dng bo ch khc tn sut dng thuc c th tng t nhng GRDF lm thay i ng k s gii phng hot cht do tng SKD ca thuc. V d nh Furosemid dng bo ch ni c SKD tng ng k (42,9%) so vi dng vin nn thng mi (Lasix (33,4%)) [24].

Thng qua gii phng thuc ti ch, GRDF lm tng cao nng thuc nim mc d dy gip tng hiu qu iu tr vim lot d dy - t trng, vim thc qun, gim nguy c ung th d dy v kim sot gii phng cc thuc khng acid [24].

GRDF c th c s dng nh cht mang vi cc thuc c ca s hp thu hp v d nh cc thuc khng virus, khng sinh, chng nm. Ngoi ra, bng vic

6

cung cp lin tc thuc ti v tr hp thu ti u, GRDF gip tng cng hiu qu s dng cc thuc c bn cht peptid v protein qua ng ung nh calcitonin, erythropoietin, vasopressin, insulin, heparin phn t lng thp, v.v[24]

Tm li, GRDF thch hp bo ch mt s loi dc cht sau [25]: - Dc cht c ca s hp thu hp ng tiu ha (v d nh L-DOPA, acid

p-aminobenzoic, riboflavin, furosemid). - Dc cht cn pht huy tc dng ti ch trong d dy (v d misoprostol,

thuc khng acid). - Dc cht khng n nh trong mi trng rut non (v d nh captopril,

ranitidin, metronidazol). - Dc cht c nh hng n h vi khun ng rut (v d nh cc thuc

khng sinh dit tr vi khun H.pylori). - Dc cht c tan km pH kim (v d diazepam, clodiazepoxid,

verapamil). - Dc cht c tnh thm km nn hn ch hp thu ng tiu ha (v d

acyclovir). 1.2.2. Cc c ch kim sot gii phng thuc ti d dy

Cho n nay c rt nhiu dng bo ch c nghin cu v pht trin vi nhiu c ch khc nhau gi li trong d dy gip tng thi gian lu ca thuc. C th k n mt s h nh sau [15], [23]: - H c t trng nh hn t trng dch v ni trn b mt dch v. - H c t trng ln hn t trng dch v lu li phn di ca d dy. - H kt dnh vi nim mc d dy. - H trng n hoc thay i hnh dng hn ch s tho rng ca h qua mn v. - H lm chm tho rng d dy bng cch dng ng thi vi thuc hoc t dc. - H kt hp ni v kt dnh, v.v 1.2.3. Mt s h kim sot gii phng thuc ti d dy 1.2.3.1. H kt dnh sinh hc (Bioadhesive drug delivery systems - BDDS) a. Khi nim kt dnh sinh hc:

Kt dnh sinh hc c th c nh ngha l trng thi gia hai vt cht m t nht mt trong hai c bn cht sinh hc c kt dnh vi nhau trong mt thi gian

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ng [13], [29]

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7

phn b thuuyn thuc nim mc, l

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mm [29] khuch tnng nhy. s khuchnh ny l: t

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11]:

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8

c cn thitc xc nh t b o lc Kqu ca cc Cc lin kc ht Vang lin kt thi lin kt rlc kt dn

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[29]

9

Bng 1. Phn loi polyme kt dnh sinh hc [11], [20], [27] Loi polyme c im V d in hnh

Polyme th h 1

Polyme cation

Kh nng KDSH do tng tc tnh in gia nhm chc mang in tch dng trong phn t vi acid sialic tch in m ca glycoprotein mng nhy.

Chitosan Trimethyl chitosan Aminodextran

Polyme anion

Kh nng KDSH do nhm carboxyl trong phn t to lin kt hydro vi nhm hydroxyl trn chui oligosaccarid ca protein cht nhy. Cc polyme anion kt dnh hiu qu hn polyme cation v polyme khng ion ha.

Carbopol CMC NaCMC Natri alginat Gm xanthan Pectin

Polyme khng ion ha

Kh nng KDSH km hn so vi polyme anion v cation. S kt dnh vi cht nhy khng ph thuc vo pH hay s c mt ca cc ion trong mi trng.

HPMC HEC PVA PVP

Polyme th h 2 Kt dnh trc tip vi b mt t bo hn l lp cht nhy bng receptor c hiu hoc lin kt cng ha tr thay v cc c ch khng c hiu nh polyme th h trc.

Chitosan thiol ha Lectin

Mt s polyme kt dnh sinh hc thng dng: Tnh cht v ng dng ca mt s polyme KDSH thng dng c th hin

trong bng 2. Bng 2. Tnh cht v ng dng ca mt s polyme kt dnh sinh hc [3],[21],[27],[32]

Polyme Tnh cht ng dng

Carbopol

Loi dc dng: 934P, 940P, 971P, 974P...

KLPT: 1106 4106 (Dalton) nht: CP934P: 29.400 39.400 cps (gel 0,5%) CP940P: 40.000 60.000 cps (gel 0,5%) Bt trng, mn, ht m, c mi c

trng, khng tan hoc rt t tan trong

Tc nhn to gel, cht nh ha, cht n nh tt, l thnh phn ph bin trong cc dng bo ch KDSH.

Kt dnh sinh hc bng lin kt hydro v lc Van der Waals gia nhm carboxyl ca Carbopol vi acid sialic ca glycoprotein mng nhy.

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nc nhng trng phng trong nc to gel, c pH acid (2,5 3,0). Trung ha vi kim to gel c nht cao hn, c hn.

Tng i bn, khng b nh hng bi s bin i ca nhit , m, cht oxy ha, c th khng li s pht trin ca vi khun.

HPMC (hydroxy

propyl methyl

cellulose)

Loi dc dng: E5, E15, E50, E4M, F50, F4M, K100, K4M, K15M, K100M

KLPT: 8,5104 15104 (Dalton) nht: 4.000cps K4M, 100.000cps

K100M (dung dch 2%). Tan trong nc lnh, hn hp methylen

clorid v isopropanol. Khng tan trong cn, cloroform v ether. n nh khi kh, dng dung dch n nh pH 3,0 11,0.

Tc nhn to gel, lm tng nht, cht n nh.

Thnh phn trong thuc m v vin nn kt dnh.

Hot tnh b mt tt, trng n trong mi trng thn nc, cc si lin kt cho trong phn t to thnh ct lu tr thuc.

Chitosan

Polyme cation c bn cht polysaccarid, c deacetyl ha mt phn t chitin.

t tan trong nc, ethanol 95% v dung mi hu c khc, tan nhanh trong cc dung dch c hoc long ca hu ht cc acid hu c v mt s acid v c (tr H2SO4 v H3PO4).

tan ph thuc vo mc deacetyl ha, mc deacetyl ha trn 85% t c tnh tan mong mun.

Tc nhn KDSH do to lin kt hydro hoc lin kt ion gia nhm amin tch in dng ca chitosan vi acid sialic tch in m ca glycoprotein mng nhy. Hn na, Chitosan cn lm tng tnh thm ca thuc qua b mt nim mc.

Khng c v t c hp thu, tng hp sinh hc tt v c kh nng phn hy sinh hc.

Tuy nhin, chitosan l mt base yu c pH khong 5,5 do khng tan trong mi trng kim v trung tnh nh rut non nn khng lm tng hp thu thuc mi trng ny. Trong trng hp ny c th s dng chitosan c trimethyl ha nhm amin tng tan ca chitosan.

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Natri alginat

Sn phm carbohydrat tinh khit c chit t rong bin bng dung dch kim long.

Bt mu trng, khng mi, khng v. Tan trong nc to dung dch keo c pH 7,2. Khng ha tan trong cc dung mi hu c v dung dch acid c pH di 3,0.

nht: 20 400cps (dung dch 1%). nht thay i ph thuc vo nhiu yu t nh pH, cc mui kim loi

An ton, khng gy kch ng, d kim, gi thnh r.

Dng lm cht nh ha, cht n nh, thnh phn trong vin nn kt dnh.

c tnh to gel tt, tng hp sinh hc tt.

Dung dch c kh nng chng vi khun v s phn hy ca enzym.

1.2.3.2. H thuc ni d dy (Floating drug delivery systems FDDS) a. Nguyn tc:

H ko di thi gian lu ti d dy bng cch thit k t trng ca h nh hn t trng ca dch v do h c th ni trn b mt dch v trong khong thi gian di m t b nh hng ca s tho rng d dy. Trong khi h ni trong d dy, dc cht s c kim sot gii phng vi tc nh gip cho vic hp thu thuc trit v ng u hn, d dng hn trong vic d on SKD v kim sot tt hn nng thuc trong mu. Sau khi gii phng ht dc cht th phn cn li ca dng thuc c tho rng khi d dy [19].

Cc yu cu chnh i vi h thuc ni l: 1. Gii phng thuc t t; 2. Duy tr t trng nh hn t trng dch v (1,004 1,01 g/cm3); 3. Hnh thnh lp hng ro gel kt dnh [25]. Ngoi ra, gi cho h ni trn b mt dch v cn yu cu lc ni phi thng c trng lc ca h. Khi , lc duy tr s ni c tnh theo phng trnh sau [24]:

F = Fni P = (Df - Ds). g.V Trong : P: trng lc ca h; Df : t trng dch v ; Ds : t trng ca h; V: th tch ca h; g : gia tc trng trng.

b. Phn loi: H ni d dy c th chia thnh 2 loi da vo c ch ni: h khng si bt

v h si bt (hoc h khng to kh v h to kh) [19]. H khng si bt:

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C ch: H khng si bt ch yu da vo c ch trng n ca polyme. Cc t

dc thng c s dng trong h ny gm c cc t dc to gel, c kh nng trng n tt nh cc dn cht ca cellulose, polysaccharid, gm hoc cc polyme to ct nh polycarbonat, polyacrylat, polymethacrylat, polystyren, chitosan, carbopol, natri alginat, v.v... Khi tip xc vi dch d dy, cc polyme s trng n h t t trng nh hn 1 cng vi lp gel bao bn ngoi. Thm vo cu trc gel ng vai tr l ngun cha dc cht gii phng ko di, qua dc cht c gii phng t t bng cch khuch tn chm qua hng ro gel [15], [19].

Phn loi: H khng si bt gm nhiu dng bo ch khc nhau nh: vin nn mt lp,

vin nn hai lp, vi cu ni, ht alginat ni, v.v[19].

Hnh 3. Cc c ch ni [24] (a) Trong h khng si bt,h hp th dch v v trng n (b) Lc ti thiu duy tr s ni (c) Trong h si bt, CO2 gii phng ra to thnh lc ni

H si bt: C ch: H s dng tc nhn to kh l natri bicarbonat kt hp vi acid citric hoc

mt khoang cha cht lng c th kh ha nhit c th. T l ti u ca acid citric v natri bicarbonat to ra kh CO2 cn thit cho s ni c nghin cu l 0,76 : 1. ng thi, trong thnh phn ca h cn cha cc polyme c kh nng trng n nh HPMC, chitosan. Khi vo n d dy, kh gii phng ra s b nht

Trng lc

Lc ni Dch

v CO2

H khng si bt H si bt

(a) (c) (b)

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trong lp gel (do polyme trng n to thnh) lm gim t trng ca h v h s ni trong d dy [25].

Phn loi: H si bt chia thnh h to kh CO2 v h cha cht lng d bay hi [25].

1.2.3.3. H c t trng ln (hay h sa lng): H c t trng ln hn t trng ca dch v (~1,004 g/cm3) nn c th c

lu li ti nhng np gp trn thnh d dy v gii phng thuc ngay ti . H c bo ch bng cch trn thm cc t dc tr nh bt st, bari sulfat, km oxid, titan oxid. Cc t dc ny gip tng t trng ca h ln n 1,5 2,4 g/cm3 [25]. 1.2.3.4. H trng n hoc thay i hnh dng

H c th trng n hoc thay i hnh dng nhanh chng khi vo d dy t c kch thc ln hn kch thc ca mn v nh cn tr s tho rng khi d dy qua mn v. H c bo ch t cc polyme c th phn hy sinh hc, da vo s hp th dch d dy ca polyme t ti kch thc mong mun [25].

1.2.3.5. H kt hp ni v kt dnh H KDSH c nhc im l d b ra tri khi d dy cng lp cht nhy.

Khi lp cht nhy b ra tri khi d dy s mang theo cc ht kt dnh xung rut non. ng thi khi c mt thc n trong d dy, s nho trn ca thc n cng nh nhu ng rut s lm gim ng k s kt dnh vi nim mc d dy [30].

Trong khi , mt nhc im ln ca h ni l phi c mt lng dch v ln h c th ni v khi d dy rng, dng bo ch s nm mn v nn s ni ca h c th b cn tr. Do , kh nng ni trong d dy c th b gii hn ch t 3 4 gi. Hn na, h ni khng phi lc no cng gii phng thuc v tr xc nh [30].

Ct lu tr thuc T dc

trng n

V polyme n hi Dch

v

Gii phng thuc

Hnh 4. S gii phng thuc t h trng n [25]

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gii quyt vn ny, nhiu nghin cu trn th gii kt hp c h kt dnh v h ni. H kt hp ny c th khc phc c nhng nhc im k trn ca c h KDSH v h ni, lm tng thi gian tip xc ca thuc vi nim mc d dy gip s hp thu thuc hiu qu hn, tng SKD ca cc thuc c ca s hp thu hp d dy v phn u rut non, gim tn sut dng thuc [30].

Zheng J. v cng s [35] bo ch vi cu ni kt dnh cha clarithromycin tng hiu qu dit H.pylori. Vi cu c bo ch bng cch phn tn clarithromycin, ethylcellulose v chitosan trong diclorometan sau cho bc hi dung mi. Tip , vi cu c bao bi alginat v phn tn trong dung dch chitosan to thnh vi ht kt dnh chitosanalginatethylcellulose (CAEMs). Kt qu nghin cu in vitro cho thy c 74% CAEMs ni trong h m acetat trong 8 gi v 90% clarithromycin c trong CAEMs gii phng t t trong 8 gi. Th nghim kt dnh in vivo cho thy c 61% CAEMs cn li trong d dy sau 4 gi. Nng clarithromycin o c trong nim mc d dy ca CAEMs cao hn hn so vi dng dung dch. Do , CAEMs c coi l h phn phi thuc y tim nng gip tng hiu qu iu tr nhim H.pylori .

Sonar G.S v cng s [31] nghin cu bo ch vin nn 2 lp kt hp ni kt dnh cha rosiglitazon maleat. Lp th nht lp lm cho vin ni gm c natri bicarbonat l tc nhn to kh, HPMC K100M l t dc to ct gi li bt kh v Starch 1500. Lp th 2 l lp gii phng ko di cha dc cht v HPMC K100M l t dc to nn hng ro gel kim sot gii phng dc cht. Kt qu th nghim in vivo trong d dy ca ngi tnh nguyn cho thy sau khi th 8 gi vin nn vn cn gi nguyn hnh dng trong d dy, chng t cc iu kin trong d dy khng nh hng n c tnh to gel ca vin.

T c th thy rng s kt hp gia h ni v h kt dnh lm gim ng k nhng nh hng ca nhiu yu t lun bin i trong d dy nh dch v, nhu ng rut, cht nhy, s c mt ca thc n, v.v.. ti kh nng lu gi thuc trong d dy. Ko di thi gian lu thuc dn n tng hp thu thuc s rt hu ch vi nhng thuc gp vn trong hp thu ng tiu ha in hnh nh ACV mt thuc va c tnh thm km li va c ca s hp thu hp d dy v on u

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rut non. Do , bo ch vin nn ACV theo hng kt hp ni v KDSH l mt hng nghin cu c trin vng nhm ci thin SKD ng ung ca thuc. 1.3. Mt s nghin cu v h kt dnh sinh hc ng tiu ha cha acyclovir 1.3.1. Nghin cu bo ch vin nn

Dias R.J. v cng s [17] tin hnh nghin cu bo ch vin nn ACV KDSH ng ung bng phng php dp thng. Th nghim c thit k theo m hnh 32 y kho st nh hng ca cc bin u vo nh Carbopol 934P v HPMC K100M ti kh nng trng n, lc kt dnh nim mc ex vivo v GPDC in vitro. Kt qu nghin cu cho thy c 9 mu vin bo ch u c kh nng GPDC ko di 12 gi. Hai loi polyme s dng u c kh nng KDSH tt trn nim mc d dy cu. Khi tng nng 2 polyme th kh nng trng n v lc KDSH ca ch phm u tng ln trong khi kh nng GPDC gim xung. Tuy nhin do trong nghin cu ny khi lng ca Carbopol 934P thay i trong phm vi hp l 50 55mg so vi HPMC K100M l 45 135 mg nn tc gi nhn thy nh hng ca HPMC K100M ti cc bin u ra chim u th hn.

Panigrahy R.N, Mahale A.M [26] tin hnh nghin cu bo ch vin nn ACV KDSH dng ct bng phng php dp thng s dng cc polyme khc nhau nh HPMC K15M, Carbopol 971P v natri alginat. Tt c cc mu vin bo ch u t cht lng tt v cng (5 6 kg/cm2), t trng (~1g/cm3), hm lng (>90%) v thi gian kt dnh ex vivo (>12h), trong Carbopol c kh nng KDSH tt hn HPMC K15M v alginat. Cng thc B8 cha 30mg Carbopol 971P v 60mg HPMC K15M c s GPDC ph hp nht vi m hnh ng hc bc khng c la chn lm cng thc ti u cho nhng nghin cu tip theo. Nghin cu n nh ca l ti u cho thy khng c s thay i ng k cc thuc tnh ha l ca dc cht cng nh kh nng KDSH v GPDC sau 90 ngy bo qun iu kin lo ha cp tc (nhit 40C v m tng i 75%).

Sapkal S.B. v cng s [28] nghin cu bo ch vin nn ni KDSH ACV dng kt hp c 3 polyme: HPMC K100M, HPMC K15M v Carbopol 934P, tc nhn to kh l natri bicarbonat kt hp vi acid citric. C 7 cng thc bo ch c t l cc polyme khc nhau u ni in vitro tt, trong cng thc 7 cha 280mg HPMC K100M 70mg HPMC K15M 100mg Carbopol 934P c thi gian

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ni lu nht ti hn 24 gi v gii phng 99,45% ACV trong 24 gi. L 7 c nghin cu n nh iu kin lo ha cp tc trong 2 thng v cho thy khng c s thay i ng k v kh nng GPDC. Do , cng thc 7 c la chn l cng thc ti u cho vin ni ko di 24 gi tip tc nhng nghin cu tip theo.

Tuncer Degim v cng s [14] bo ch vin nn ACV kt dnh nim mc ming vi cc thnh phn: CP934P, polycarbophil (PCa) v natri taurocholat (ST). Kh nng kt dnh nim mc ming ca cc mu vin bo ch c nh gi trn nim mc ming b bng thit b o lc ko Instron model 4301. Kt qu nghin cu cho thy khi tng lng CP934P trong cng thc th kh nng kt dnh nim mc tng, ngc li kh nng ny gim khi tng lng PCa. Vic thm ST vo cng thc c tc dng lm tng s hp thu ca ACV qua nim mc. Qua kho st kh nng GPDC trong mi trng dch nc bt nhn to, cng thc thch hp nht cho cc th nghim tip theo c thnh phn: CP934P (64,4%), PCa (34,4%), ST (0,8%). Cng thc ny GPDC theo m hnh ng hc bc 0 vi tc 9,708 mg/h, thi gian GPDC ko di 6 gi. Kt qu xc nh nng thuc trong nc bt v cc thng s dc ng hc trn ch, so snh vi thuc tim tnh mch v vin nn ung trn th trng (Hernovir) cho thy vin nn kt dnh nim mc ming bo ch c tim nng trong iu tr bnh Herpes c ti ch ming v ton thn. 1.3.2. Nghin cu bo ch vi cu

Dhaliwal S. v cng s [16] nghin cu bo ch vi cu kt dnh ACV s dng cc polyme KDSH nh: Chitosan, Chitosan thiol ha, Carbopol 71G v Methocel K15M. Cc vi cu chitosan v chitosan thiol ha c bo ch bng k thut nh ha kt hp vi tc nhn lin kt cho l glutaraldehyd. Cc vi cu Carbopol 71G v Methocel K15M c bo ch bng phng php phun sy. Cc mu vi cu c ng nang v nh gi kh nng GPDC. Thi gian ACV gii phng 75% t nang gelatin cha vi cu Chitosan, Chitosan thiol ha, Methocel K15M v Carbopol 71G tng ng l 5,0; 9,5; 4,0 v 5,5 gi trong khi nang cha bt ACV c ha tan sau 1 gi l 90,5 3,6%. Thi gian kt dnh ca cc mu vi cu trn rut ln c xp theo th t: Chitosan thiol ha (8,0 gi) > Chitosan (3,1 gi) > Carbopol 71G (1,1 gi) > Methocel K15M (0,2 gi). Nghin cu dc ng hc trn chut cho thy vi cu cha Chitosan thiol ha c AUC0-24 (1090 51

17

ng.h/ml) cao gn gp 4 ln so vi dng dung dch ACV (281 28 ng.h/ml). Do , vi cu ACV bo ch t Chitosan thiol ha l mt hng kh thi nhm ko di thi gian lu ca thuc ti v tr hp thu, ci thin ng k SKD ng ung ca ACV.

trong nc, vi cu ACV KDSH cng bc u c nghin cu bo ch bng phng php bc hi dung mi. Kt qu kho st cho thy cc mu vi cu ACV bo ch c kch thc phn b kh ng u trong khong 0,60 1,18 mm, hm m tng i thp (0,8 1,2 %), khi lng ring biu kin dao ng trong khong 0,60 0,64g/ml, kh nng trng n kh tt t 1,61 1,95 ln v kh nng gii phng ACV ko di trn 8 gi. Nghin cu kh nng kh nng kt dnh trn nim mc ng tiu ha chut ca vi cu bo ch so vi mu vi cu ACV-ms (khng KDSH, bo ch cng phng php) cho thy ti tn 6 gi sau khi ung, vn cn trn 26% lng vi cu cn c kt dnh trn nim mc d dy. Tng s vi cu cn kt dnh trn nim mc ng tiu ha ti v tr hp thu ca cc mu nghin cu cc thi im nghin cu u cao hn nhiu so vi mu ACV-ms. V vy c th khng nh vic kt hp Carbopol 934P vi EC t l thch hp gp phn kim sot kh nng GPDC ng thi ko di thi gian lu ca thuc trn nim mc d dy v rut non [7]. 1.3.3. Nghin cu bo ch h nano

Bhosale U.V. v cng s [12] nghin cu bo ch h nano KDSH cha ACV bng phng php bc hi dung mi. Cc polyme KDSH c s dng trong nghin cu gm acid poly lactic-co-glycolic (PLGA, 50:50) v polycarbophil (Noveon AA-1), Pluronic F68 c s dng lm t dc n nh. Tc gi thit k 23 th nghim vi cc bin u vo gm lng PLGA, Pluronic F68, polycarbophil. Cc bin u ra bao gm kch thc tiu phn, hiu sut ch to v phn trm thuc gii phng sau 12 gi. Kt qu nghin cu cho thy t l thuc/polyme v nng cht n nh c nh hng ti kch thc tiu phn v hiu sut a ACV vo h nano KDSH. Qu trnh GPDC t cc mu bo ch tun theo nh lut Fick v ng hc bc khng. Kh nng KDSH in vitro trn rut non chut ca h nano tng theo t l polycarbophil trong cng thc. Kt qu nghin cu cn cho thy hiu sut np ACV vo h nano c th nh hng ti kh nng kim sot gii phng thuc ko di.

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PHN 2. I TNG V PHNG PHP NGHIN CU 2.1. Nguyn liu v phng tin nghin cu 2.1.1. Nguyn vt liu nghin cu

Bng 3. Nguyn vt liu nghin cu STT Nguyn liu Ngun gc TCCL

1 Acyclovir Trung Quc USP 2 Carbopol 934P Php USP 3 Carbopol 940P Php USP 4 HPMC K4M M TCCS 5 HPMC K100M M TCCS 6 Natri alginat Trung Quc BP 7 Natri hydrocarbonat Trung Quc BP 8 Lactose Trung Quc Eur.Ph 9 Avicel PH101 Trung Quc USP

10 Talc Trung Quc BP 11 Magnesi stearat Php Eur.Ph 12 Aerosil Trung Quc USP 13 Acid clohydric Trung Quc TKHH 14 Natri hydroxyd Trung Quc TKHH

2.1.2. Phng tin nghin cu - Cn k thut SARTORIUS. - Cn phn tch SARTORIUS. - T sy BINDER, HERAEUS. - My siu m ULTRASONIC LC 60H. - My dp vin KORSCH EKO. - My o m SARTORIUS. - My o khi lng ring biu kin ERWEKA SVM. - My o trn chy ERWEKA GWF. - My th ha tan ERWEKA DT 600. - My o quang ph UV VIS HITACHI U1800. - My o pH EUTECH INSTRUMENTS pH 510.

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- My ht m EDISON ED 12B. - Ry 180, 250 m. - My o lc gy v vin ERWEKA TBH 200. - Thit b nh gi kh nng KDSH t ch to t cn k thut.

2.1.3. ng vt th nghim ng vt th nghim c chn l th trng trng thnh, kho mnh, ging

c, c trng lng t 2,0 - 2,5 kg, c nui dng trong iu kin dinh dng y , c kim sot, b i qua ngy v c ung nc t do trc khi tin hnh th nghim. 2.2. Phng php nghin cu 2.2.1. Xy dng ng chun acyclovir trong mi trng acid clohydric 0,1M

Cn chnh xc 100mg acyclovir cho vo bnh nh mc 100ml, thm 60ml dung dch HCl 0,1M v siu m khong 1 gi ha tan hon ton. Thm dung dch HCl 0,1M va n vch, lc u v lc. Sau pha long thnh cc dung dch c nng ln lt 4, 6, 8, 10, 12, 14 g/ml. Tin hnh o mt quang cc dung dch trn bc sng 252 nm.

T kt qu thu c xy dng phng trnh hi quy thc nghim, v th biu th s tng quan gia mt quang v nng acyclovir. 2.2.2. Phng php bo ch vin nn acyclovir kt dnh sinh hc

Vin nn ACV KDSH c bo ch bng phng php dp thng vi cc thnh phn d kin:

Acyclovir (200mg) Polyme KDSH: CP934P, CP940P, HPMC K100M, HPMC K4M,

natri alginat.

Natri hydrocarbonat Lactose Avicel PH101 Aerosil Talc Magnesi stearat

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Cc bc tin hnh bo ch vin nn ACV KDSH c th hin trong hnh 5.

2.2.3. nh gi mt s ch tiu cht lng ca bt dp vin Xc nh khi lng ring biu kin v ch s nn ca khi bt Th tch biu kin ca khi bt c o trn my ERWEKA SVM. Mt

lng bt c khi lng xc nh (m) c cho vo ng ong th tch hnh tr. Th tch khi bt ban u l V0. Sau ng ong c g cho n khi th tch khng thay i th c th tch cui cng l V.

Khi lng ring ca khi bt ban u: d0 = (g/cm3)

Khi lng ring biu kin ca khi bt sau khi g: d = (g/cm3)

Ch s nn ca khi bt: CI (%) = 100 Xc nh trn chy Tc chy ca khi bt c o trn my ERWEKA GWF, vi ng

knh l phu 12 mm. Tc chy c tnh theo cng thc: V = tg

Trong : V: tc chy (g/giy)

Nghin, ry # 180, cn - Khi lng: 700mg - Lc gy v vin: 11 13 kp - Kch thc: 18 9 3 (mm)

Hnh 5. S cc giai on bo ch vin nn acyclovir kt dnh sinh hc

Dp vin

Trn u

Chun b dc cht, t dc

Ry # 250, cn

Trn bt kp T dc trn

21

: gc gia ng thng biu din s ph thuc ca khi lng bt chy theo thi gian v trc honh (trc thi gian)

Xc nh hm m theo phng php mt khi lng do sy kh Tin hnh trn cn xc nh m nhanh SARTORIUS. Cn chnh xc

khong 1g bt, ri u vo a cn, t nhit 105C, theo di v c kt qu. 2.2.4. nh gi mt s ch tiu cht lng ca vin nn bo ch

Kh nng trng n Kh nng trng n ca vin nn bo ch c nh gi thng qua xc nh

lng nc c hp th trong vin bng phng php phn tch trng lng: vin thuc c cho vo gi lm bng thp khng g c kch thc mt li 381m, ng knh si ry l 254 m v cn xc nh khi lng ban u ca vin nn (W0). Sau gi thuc c nhng vo trong 100 ml dung dch acid clohydric 0,1M nhit 37 1oC. Sau tng khong thi gian nht nh (10 pht) gi thuc c ly ra, dng giy lc thm ht phn nc trn b mt vin v cn xc nh khi lng (Wt). Mc trng n ca vin thuc (S) c tnh theo cng thc [6]:

Kh nng kt dnh sinh hc in vitro Thit b dng xc nh lc kt dnh sinh hc c m t trong hnh 6. - X l nim mc d dy th: Sau khi git th bng cch bm khng kh

vo tnh mch vnh tai, ngay lp tc phu thut ct ly d dy th. em lm sch b mt nim mc d dy bng dung dch nc mui sinh l sao cho khng lm mt lp cht nhy trn b mt. Nim mc c s dng ngay sau khi x l, ct thnh nhng ming c din tch 4cm2 ( 22cm).

- nh gi lc KDSH: Gn c nh nim mc ln gi , lm t nim mc bng mt th tch chnh xc 0,1 ml dung dch acid hydrocloric 0,1M sau t vin thuc ln nim mc. Tc ng ln vin thuc mt lc 200g trong 1 pht. Sau iu chnh nc trn buret nh xung cc nha vi tc 3ml/pht cho n khi vin thuc b tch ra khi b mt nim mc di tc ng ca trng lng nc.

100o

ot

WWWS

Gh

n

0,1

trm

hi li khi lnh lc KDSc tnh theo

Trong

H

Kh Vin th

M. Thi gng c tt mi trn

Tin h nhTin h

C

lng ncSH ca vino cng thc

: F: Lc K m: Kh S: Din

bng ph

Hnh 6. Thinng ni in

huc c gian t khi nh l thi g

ng c tnhng u khnh theo ch

h lng: nh theo ch

Cc nc

Buret

c (chnh l

n thuc. Lc:

F

KDSH tnh

i lng nn tch b mhn mm Im

t b nh gn vitro

th vo cth n kh

gian tim th l thi giahi lng: uyn lun v

uyn lun v

th tch nc KDSH t

F = 0,00981

h cho 1cm2 (

c (g) mt vin (cmmage J 1.46

gi lc kt

c nc chhi vin thu

ng (tlag) v

an ni (tni)

vin nn ph

vin nn acy

22

c trn butnh cho 1c

1

(N/cm2)

m2) c q.

dnh sinh h

ha 100ml uc bt u tng thi g[32].

h lc 11.3 c

yclovir ca

m

S

uret nh cm2 din t

qut hnh n

hc t ch t

dung dch u ni ln trgian vin th

ca DVN

a DVN IV

Gi

V

xung) ch b mt

nh v xc

to

acid clohy

rn b mt huc ni tr

IV [4].

V [4].

i gn nimVin thuc

xc vin

nh

ydric

mi

n b

m mc

23

nh gi ho tan ca vin ACV KDSH Kh nng gii phng dc cht ca vin ACV bo ch c tin hnh

trong iu kin sau: - Thit b: Cnh khuy. - Tc khuy: 50 1 vng/pht. - Mi trng ho tan: 900 ml HCl 0,1M. - Nhit mi trng ho tan: 37 0,5oC. - Thi gian th nghim: 8h. - Ly mu vo cc thi im: 1h, 2h, 4h, 6h, 8h.

nh lng dc cht gii phng bng phng php o mt quang bc sng 252 nm. Tnh lng dc cht gii phng cn c vo mu chun c nng khong 10 g/ml.

Cch tnh kt qu: Nng ACV cha hiu chnh ln ht th n:

o

onono D

CDC

Trong : Dno: hp th ca dung dch th Co: Nng dung dch chun Do: hp th ca dung dch chun : pha long ( = 5; 12,5; 25)

Nng ACV sau khi hiu chnh trong mu ln ht th n c tnh theo cng thc Nelson:

1

1

n

ii

onon CV

VCC

Trong : Cn: Nng hiu chnh ln ht th n (g/ml) Cno: Nng cha hiu chnh ln ht th n (g/ml) Vo: Th tch dch ho tan ht (Vo = 10ml) V: Th tch mi trng ho tan (V = 900 ml) Ci: Nng hiu chnh ln ht th i (g/ml)

24

Phn trm ACV (C%) ho tan ti thi im t c tnh theo cng thc:

1001000900%

m

CC n

Trong : Cn: Nng hiu chnh ln ht th n m: Lng ACV trong vin (mg)

2.2.5. Phng php thit k th nghim v x l s liu Cc kt qu trong kha lun c x l theo phn tch thng k v trnh

by di dng: Gi tr trung bnh lch chun vi n l s ln lp li th nghim. Thit k th nghim Phn mm Modde 8.0 c s dng thit k th nghim theo m hnh mt

hp t ti tm rt gn. Ti u ho cng thc nh hng ca cc bin u vo v cng thc ti u c xc nh da trn

mng thn kinh nhn to bng phn mm INForm 3.1. So snh hai th gii phng dc cht in vitro f2 c tnh theo cng thc:

10011lg50

5.0

1

22

n

iii TRn

f

Trong : f2: Ch s biu hin cho s ging nhau i: s th t im ly mu n: s im ly mu Ri v Ti: % acyclovir gii phng ti thi im i ca vin i chiu v vin bo ch c.

Gi tr f2 trong khong t 50 100 th hai th c coi l ging nhau. Gi tr f2 cng ln th hai th cng ging nhau.

25

PHN 3. KT QU THC NGHIM V NHN XT 3.1. Kho st mi tng quan gia nng dung dch v mt quang

Tin hnh pha cc dung dch ACV c nng t 4 g/ml n 14 g/ml v o mt quang cc dung dch ny bc sng 252 nm. Kt qu th hin bng 4 v hnh 7.

Bng 4. Mt quang (D) v nng (C) ca dung dch acyclovir

Hnh 7. th biu din s ph thuc gia mt quang (D) vi nng

(C) ca dung dch acyclovir trong mi trng HCl 0,1M Nhn xt: Gi tr R2 xp x bng 1, chng t trong mi trng HCl 0,1M,

mt quang ph thuc tuyn tnh vo nng ACV trong khong kho st. Chun b 2 mu vin: Mu cha ACV v mu khng cha ACV (mu

placebo) vi cng t l cc t dc khc. - Mu cha ACV: Cn chnh xc mt lng bt vin tng ng khong

100mg ACV sau pha dung dch ACV trong HCl 0,1M c nng khong 10 g/ml. o mt quang dung dch ny bc sng 252 nm thu c gi tr DACV.

- Mu placebo: Tin hnh song song v tng t nh mu cha ACV. o mt quang dung dch ny bc sng 252 nm thu c gi tr D0.

y = 0,0386x + 0,0560R = 0,9993

00.10.20.30.40.50.60.7

0 2 4 6 8 10 12 14 16

Mt

qua

ng (D

)

Nng (g/ml)

C (g/ml) 4 6 8 10 12 14 D 0,215 0,281 0,367 0,441 0,519 0,598

26

Kt qu o mt quang ca mu cha ACV v mu placebo c th hin trong bng 5.

Bng 5. Mt quang ca mu cha acyclovir v mu placebo STT 1 2 3 4 5 6 Trung bnh DACV 0,431 0,437 0,435 0,440 0,432 0,438 0,436 0,004

D0 0,004 0,004 0,003 0,004 0,003 0,003 0,0035 0,0005

(D0/DACV)100 (%)

0,928 0,915 0,690 0,909 0,694 0,684 0,803 0,125

T kt qu trn ta thy t l < 1% chng t cc t dc trong cng thc khng c nh hng n s hp th mt quang ca ACV trong mi trng HCl 0,1M bc sng 252 nm.

Kt lun : C th s dng phng php o quang bc sng 252 nm trong mi trng HCl 0,1M nh lng ACV trong mu th v trong mi trng ho tan.

3.2. Nghin cu phng php bo ch v cc t dc c bn ca vin nn acyclovir kt dnh sinh hc 3.2.1. Nghin cu phng php bo ch Tin hnh bo ch vin nn 700mg khng cha dc cht vi tng polyme KDSH (CP934P, CP940P, HPMC K100M, HPMC K4M, natri alginat) t l 20%, cc thnh phn t dc khc khng thay i (lactose 20%, magnesi stearat 2%, Aerosil 1%, Talc 1%, Avicel 56%). Cc mu vin c bo ch song song bng 2 phng php dp thng v to ht t. Phng php to ht t:

- Do trong cng thc phi s dng mt t l ln polyme KDSH c kh nng trng n mnh trong mi trng thn nc nn ngay t u nh hng dng cc t dc dnh khan nc nh isopropanol, ethanol. Tuy nhin do CP934P v CP940P c th trng n ngay c trong cc t dc dnh khan nc ny lm bt bt dnh v vn cc do kh to ht v khng xt c qua ry.

27

- Vi natri alginat c th to c ht nhng ht khng chc, sau khi sy bay hi ht dung mi th ht b b ra kh lin kt.

- Trong s cc polyme KDSH trn ch c HPMC K100M v HPMC K4M c th to c ht. Tuy nhin, vin nn bo ch bng phng php to ht t vi t dc HPMC K100M v HPMC K4M c b mt x x, khng nhn bng. Khi nh gi kh nng trng n th vin c hin tng r ra nhanh chng ch sau 10 pht v vy kh c th p ng c yu cu gii phng ACV ko di. Phng php dp thng: Vi mu vin cha CP934P, CP940P:

- Tc chy ca khi bt thp do Carbopol d ht m v khng phi l t dc dng cho dp thng. Khi dp vin bt d b dnh chy ci. Khi lng gia cc vin dao ng nhiu mc d vn t yu cu v ng u khi lng.

- Hnh thc: B mt vin b bong, khng nhn bng. Vi mu vin cha HPMC K4M, HPMC K100M:

- Khi bt kp trn chy tt, khi dp vin t yu cu v ng u khi lng do HPMC t ht m v l t dc dng cho dp thng.

- Hnh thc: B mt vin bng p. Vi mu vin cha natri alginat: Khi bt kp trn chy tt, t yu cu v

ng u khi lng nhng b mt vin b bong, khng nhn bng. Kt lun: Bo ch vin nn ACV KDSH bng phng php to ht t gp nhiu kh khn. Trong khi , phng php dp thng c nhiu u im l t cng on, tit kim c mt bng, thit b v thi gian sn xut, trnh c tc ng ca nhit n polyme KDSH v dc cht. Tuy nhin, phng php dp thng ch c th p dng khi chn c t l polyme thch hp va p ng ko di GPDC v kh nng KDSH va m bo c trn chy ca khi bt kp. ng thi cn phi hp vi cc t dc dp thng v t dc trn chy t l thch hp. V vy, chng ti quyt nh la chn phng php dp thng bo ch vin nn ACV KDSH c kho st t l polyme v cc t dc khc m bo trn chy ca khi bt v ng u khi lng vin.

28

3.2.2. La chn cc t dc c bn bo ch vin nn acyclovir kt dnh sinh hc La chn t dc trn chy: Trn c s kho st s b vic s dng mt s t dc trn cc t l khc

nhau v tham kho cc nghin cu trc v vin nn ACV KDSH [6], chng ti nhn thy m bo kh nng trn chy ca khi bt kp cn kt hp cc t dc trn: Mg stearat, Talc, Aerosil t l 2% Mg stearat, 1% Talc, 1% Aerosil khi bt kp trn chy tt m bo ng u khi lng vin. V vy, t l cc t dc trn nh trn c c nh trong tt c cc cng thc kho st.

La chn polyme kt dnh sinh hc: la chn polyme KDSH, tin hnh bo ch cc mu vin nn cha acyclovir bng phng php dp thng vi cc thnh phn th hin trong bng 6.

Bng 6. Thnh phn ca vin nn c bo ch vi cc polyme khc nhau CT vin

Polyme KDSH Lng t dc (mg)

Polyme Lactose ACV Mg stearat Talc Aerosil Avicel A CP934P 140 100 200 14 7 7 232 B CP940P 140 100 200 14 7 7 232 C HPMC K100M 140 100 200 14 7 7 232 D HPMC K4M 140 100 200 14 7 7 232 E Natri alginat 140 100 200 14 7 7 232

nh gi mt s c tnh ca cc mu vin bo ch thu c kt qu nh sau: nh gi kh nng trng n ca cc mu vin c th hin trong bng

7 v hnh 8. Bng 7. Mc trng n ca cc mu vin theo thi gian

Mu vin

Mc trng n (%) 10 pht 20 pht 30 pht 40 pht 50 pht 60 pht

A 30,61 55,73 70,22 82,68 93,74 101,52 B 35,62 64,78 77,21 89,26 98,51 109,32 C 25,64 35,58 38,99 44,39 - - D 22,34 29,65 32,75 37,88 - - E 15,73 16,82 17,03 - - -

29

Hnh 8. Kh nng trng n ca vin bo ch vi cc polyme khc nhau Nhn xt: - Mu vin cha Carbopol c kh nng trng n mnh nht, vin cha

CP940P trng n mnh hn vin cha CP934P. - Mu vin cha HPMC K100M v K4M c kh nng trng n km hn

CP, sau 40 pht gn nh khng trng n na. - Mu vin cha natri alginat trng n km nht, sau 30 pht khng trng

n na. nh gi kh nng gii phng dc cht v kt dnh sinh hc ca cc mu vin:

tnh c lc KDSH ca mu vin, s dng phn mm Image J 1.46 xc nh din tch b mt vin nn thu c kt qu: S = 1,59 0,05 cm2 (n = 6).

Bng 8. Kt qu th GPDC v KDSH ca cc mu vin (n=3) Mu vin

Phn trm ACV gii phng (%) Lc KDSH (N/cm2) 1 gi 2 gi 4 gi 6 gi 8 gi

A 15,47 2,18 26,05 3,08

38,21 2,71

47,57 2,35

55,12 2,12

0,56 0,07

B 13,77 2,21 22,43 1,37

31,63 3,28

39,66 3,16

47,98 4,68

0,58 0,06

C 16,28 4,36 27,42 1,57

39,54 3,15

51,35 2,32

63,23 1,79

0,38 0,09

D 17,48 1,81 29,2

2,24 41,84 3,04

54,68 2,85

67,52 2,54

0,36 0,11

E 13,85 2,17 23,53 2,45

35,71 2,47

42,86 3,01

50,93 3,12

0,31 0,05

0

20

40

60

80

100

120

0 20 40 60 80

Mc

tr

ng n (

%)

Thi gian (pht)

Mu AMu BMu CMu DMu E

30

Hnh 9. th gii phng dc cht ca cc mu vin bo ch vi polyme khc nhau Nhn xt:

- V kh nng KDSH: Mu vin cha CP934P v CP940P c lc KDSH ln nht, cc mu vin

cha HPMC K100M v K4M c lc KDSH km hn CP. Mu vin cha natri alginat c lc KDSH thp nht.

- V kh nng GPDC: Cc polyme KDSH u ko di qu trnh GPDC do to ra lp gel bao xung

quanh b mt vin khin mi trng thm vo vin kh hn v khuch tn dc cht ra ngoi chm hn. Trong , mu vin cha CP940P lm gim tc GPDC nhiu nht do CP940P c nht cao nht nn to ra lp gel c cn tr GPDC. Mu vin cha CP934P v natri alginat cng lm gim tc GPDC ng k. Mu vin cha HPMC K100M v HPMC K4M t cn tr GPDC hn so vi CP v natri alginat.

Tm li: Qua kt qu kho st s b ta nhn thy trong cc polyme kho st, CP934P v CP940P c kh nng trng n v KDSH tt nht nhng nu ch dp vin cha CP th bt trn chy km, d dnh chy ci, b mt vin xu, d bong mt, ng thi kh nng GPDC km. Khi so snh 2 loi CP kho st, mu vin cha CP940P c kh nng trng n v KDSH cao hn CP934P khng ng k nhng li ko di qu trnh GPDC hn. Do , CP934P c chn l polyme KDSH thun li cho kh nng GPDC.

Bn cnh , cc mu vin bo ch vi HPMC K100M v HPMC K4M c kh nng trng n v kt dnh km hn CP nhng b mt vin li bng p, bt

0

10

20

30

40

50

60

70

80

0 2 4 6 8 10

% A

CV

gii

ph

ng

Thi gian (gi)

Mu AMu BMu CMu DMu E

31

trn chy tt, khng b dnh chy ci. Mu vin cha natri alginat c kh nng trng n, kt dnh km nht v tc GPDC thp. Do chng ti quyt nh phi hp CP934P vi HPMC K100M hoc HPMC K4M tn dng u im v khc phc nhc im ca nhng polyme ny.

la chn polyme kt hp vi CP934P ng thi kho st s b t l polyme trong vin nn ACV, tin hnh bo ch cc mu vin vi thnh phn th hin trong bng 9.

Bng 9. Thnh phn cc cng thc kho st CT vin

CP 934P (mg)

HPMC K100M

(mg)

HPMC K4M (mg)

Lactose (mg)

Avicel (mg)

ACV (mg)

T dc trn (mg)

M1 50 50 - 100 272 200 28 M2 50 - 50 100 272 200 28

M3 80 20 - 100 272 200 28

M4 80 50 - 100 242 200 28

M5 60 20 - 100 292 200 28

Sau khi dp vin tin hnh nh gi kh nng trng n, KDSH v GPDC ca cc mu vin thu c kt qu nh sau:

Kh nng trng n: Kt qu nh gi kh nng trng n ca cc mu vin c th hin trong bng 10 v hnh 10.

Bng 10. Mc trng n ca cc mu vin kho st Mu vin

Mc trng n (%) 10 pht 20 pht 30 pht 40 pht 50 pht 60 pht

M1 27,64 41,85 53,72 63,98 72,81 80,92 M2 23,11 37,92 46,51 57,69 67,43 76,84 M3 35,23 52,27 63,83 75,42 88,39 95,84 M4 38,44 55,87 66,72 80,84 92,61 102,92 M5 21,54 33,75 43,73 52,69 64,24 72,48

32

Hnh 10. Kh nng trng n ca cc mu vin theo thi gian

Nhn xt: - Mu vin M1 c mc trng n cao hn mu M2 chng t khi phi

hp vi cng t l CP934P th HPMC K100M trng n mnh hn HPMC K4M do nht ca K100M ln hn nhiu so vi K4M.

- Mu M4 c khi lng 2 polyme nhiu nht c MTN cao nht trong khi mu M5 c khi lng 2 polyme t nht c MTN thp nht. Khi tng lng CP934P v HPMC K100M trong CT th MTN tng ln.

Kh nng GPDC v KDSH: Kt qu nh gi kh nng GPDC v KDSH ca cc mu vin c th hin trong bng 11.

Bng 11. Kh nng GPDC v lc KDSH ca cc mu vin kho st (n=3) Mu vin

Phn trm ACV gii phng (%) Lc KDSH (N/cm2) 1 gi 2 gi 4 gi 6 gi 8 gi

M1 13,82 2,18 25,64 1,82

39,18 3,54

54,25 4,21

67,48 2,19 0,38 0,09

M2 15,57 3,05 28,18 3,32

43,68 2,75

57,51 3,46

70,54 3,92

0,34 0,06

M3 17,23 2,25 32,42 4,15

48,62 1,91

62,87 1,28

76,65 2,54

0,43 0,08

M4 11,26 3,26 19,84 3,22

32,15 2,32

43,72 2,64

56,89 3,06 0,45 0,10

M5 16,14 3,28

33,58 2,84

53,62 1,65

71,24 2,47

85,36 2,18

0,41 0,07

0

20

40

60

80

100

120

0 20 40 60 80

Mc

tr

ng n (

%)

Thi gian (pht)

Mu M1Mu M2Mu M3Mu M4Mu M5

33

Hnh 11. th gii phng dc cht theo thi gian ca cc mu vin

Nhn xt: V kh nng GPDC: - Mu M2 c kh nng GPDC cao hn mu M1 i cht do HPMC K4M c

nht thp hn HPMC K100M nn t cn tr GPDC hn. - Khi tng khi lng CP934P v HPMC K100M trong CT th tc

GPDC gim r rt. V d nh mu M3, M4 tng khi lng HPMC K100M (20 50 mg), % ACV gii phng sau 8h gim nhiu (76,65 56,89%). mu M1, M4 tng khi lng CP (50 80mg), % ACV gii phng sau 8h cng gim i ng k (65,48 56,89%).

V lc KDSH: - Mu M1 c lc KDSH cao hn mu M2 chng t HPMC K100M c kh

nng kt dnh mnh hn HPMC K4M. - Lc KDSH ca vin tng theo s tng t l CP934P v HPMC K100M.

Khi tng khi lng CP934P th lc KDSH tng ln ng k, v d mu M1, M4 tng khi lng CP934P (50 80 mg) lc KDSH tng ln nhiu (0,38 0,45 N/cm2). mu M3, M4 tng khi lng HPMC K100M (20 50 mg) lc KDSH tng ln t hn khi tng CP934P (0,43 0,45 N/cm2). Chng t CP934P l polyme c nh hng ln hn n lc KDSH.

Kt lun: So snh 2 mu vin cha cng mt lng CP934P th mu vin cha HPMC K100M c kh nng trng n v kt dnh tt hn mu vin cha HPMC K4M, kh nng GPDC c km hn nhng khng nhiu. V vy, chng ti quyt nh la chn hn hp 2 polyme CP934P v HPMC K100M tip tc tin hnh kho st v ti u ha cng thc vin nn ACV KDSH.

0102030405060708090

0 2 4 6 8 10

% A

CV

gii

ph

ng

Thi gian (gi)

Mu M1Mu M2Mu M3Mu M4Mu M5

34

La chn t dc to kh: m bo duy tr tt kh nng lu gi gip cho hp thu thuc n nh hn, chng ti nh hng nghin cu bo ch vin nn acyclovir va c kh nng KDSH va ni trong d dy. T dc to bt kh l natri hydrocarbonat c la chn nghin cu. Tin hnh kho st mt s cng thc vin nn c t l natri hydrocarbonat thay i xem xt nh hng ca t dc ny n mt s c tnh ca vin ACV thu c cc kt qu nh sau:

Bng 12. Lc KDSH v mc trng n ca cc mu vin c t l natri hydrocarbonat khc nhau

CT Lng

NaHCO3 (mg)

Mc trng n (%) Lc KDSH

(N/cm2) 10 pht 20 pht 30 pht 40 pht 50 pht 60 pht

M1 0 27,64 41,85 53,72 63,98 72,81 80,92 0,38 0,09 M6 70 29,12 44,28 55,62 67,31 78,02 86,14 0,40 0,08 M7 100 33,45 46,75 59,31 71,14 82,56 88,52 0,41 0,06

(T l cc thnh phn khc c gi c nh l: 200mg ACV, 50mg CP934P, 50mg HPMC K100M, 100mg lactose, 28mg t dc trn, Avicel PH101 v 700mg).

Nhn xt: Khi tng t l natri hydrocarbonat trong CT th kh nng trng n v KDSH ca vin u tng ln. iu ny c th gii thch l do natri hydrocarbonat trung ha CP934P lm tng nht ca CP934P nn CP trng n tt hn, to ra lp gel c hn lm tng kh nng KDSH.

nh gi kh nng GPDC v kh nng ni ca cc mu vin: Bng 13. Kh nng GPDC v kh nng ni ca cc mu vin

thay i t l natri hydrocarbonat (n = 3) Mu vin

Kh nng ni % ACV gii phng tlag (pht) tni (gi) 1 gi 2 gi 4 gi 6 gi 8 gi

M1 - - 13,82

2,18 25,64 3,82

39,18 3,54

54,25 4,21

67,48 2,19

M6 21,67 3,51 > 12

11,31 2,24

22,52 2,79

34,84 4,28

49,14 2,07

62,72 3,85

M7 13,33 4,28 > 12

11,24 3,09

21,56 1,96

32,58 2,64

47,21 3,56

59,63 2,71

(Du -biu th vin khng ni)

35

Nhn xt: - V kh nng ni: Mu M1 khng c natri hydrocarbonat nn khng ni.

Khi tng lng natri hydrocarbonat trong CT th thi gian tim tng gim i do natri hydrocarbonat to ra nhiu kh CO2 hn nn vin ni nhanh hn. Cc mu M6, M7 u ni hn 12 gi m khng c hin tng r.

- V kh nng GPDC: Khi tng t l natri hydrocarbonat trong CT th tc GPDC gim i nhng khng nhiu. Cc mu M1, M6, M7 c t l natri hydrocarbonat tng dn th kh nng GPDC gim dn. Do natri hydrocarbonat trung ha CP934P lm tng nht ca CP to ra lp gel c hn cn tr khuch tn dc cht ra ngoi.

Kt lun: Vic phi hp natri hydrocarbonat vo thnh phn vin nn ACV KDSH ng vai tr l t dc to kh CO2 trong mi trng acid clohydric gip cho vin ni. Mt khc, natri hydrocarbonat trung ha CP934P lm tng nht ca CP934P nn tng kh nng trng n v KDSH ca CP934P cn kh nng GPDC gim nhng khng ng k. V vy, natri hydrocarbonat c chn l t dc to bt kh bo ch vin nn ACV kt hp ni KDSH. 3.3. Xy dng cng thc bo ch vin nn acyclovir kt dnh sinh hc 3.3.1. Quy hoch thc nghim

Trong qu trnh kho st s b nhn thy cc thnh phn c bn trong cng thc nh CP934P, HPMC K100M, natri hydrocarbonat u nh hng n cc c tnh ca vin ACV KDSH. V vy, chng ti tin hnh thit k th nghim kho st nh hng ca cc t dc trn n kh nng chy ca khi bt kp, kh nng GPDC, kh nng KDSH, ni v trng n ca vin nn bo ch.

Cc thng s gi c nh bao gm: Acyclovir: 200mg Lactose: 100mg T dc trn: 28mg (14mg Magnesi stearat, 7mg Aerosil, 7mg Talc) Khi lng vin: 700mg Lc gy v vin: 11 13kp

36

Cc bin c lp: Cn c vo kt qu nghin cu s b, cc bin c lp c la chn vi cc

mc th hin trong bng 14. Bng 14. Cc bin c lp

STT Bin c lp Gii hn di

Gii hn trn

Mc c bn

1 Carbopol 934P (mg) 50 90 70 2 HPMC K100M (mg) 10 40 25 3 Natri hydrocarbonat (mg) 85 115 100

Cc bin ph thuc: Bng 15. Cc bin ph thuc

Bin ph thuc K hiu Yu cu Phn trm Acyclovir gii phng sau 1h (%) Y1 15 25 Phn trm Acyclovir gii phng sau 2h (%) Y2 25 40 Phn trm Acyclovir gii phng sau 4h (%) Y4 45 60 Phn trm Acyclovir gii phng sau 6h (%) Y6 65 80 Phn trm Acyclovir gii phng sau 8h (%) Y8 > 85

Lc KDSH (N/cm2) KDSH > 0,30 S dng phn mm Modde 8.0 thit k th nghim theo m hnh mt hp

t ti tm. Cc cng thc thc nghim c trnh by bng 16. Bng 16. Cc cng thc thc nghim

CT vin

HPMC K100M

(mg)

CP934P (mg)

NaHCO3(mg)

Lactose (mg)

Avicel PH101 (mg)

ACV (mg)

Mg stearat (mg)

Talc(mg)

Aerosil(mg)

CT1 10 50 85 100 227 200 14 7 7 CT2 10 90 85 100 187 200 14 7 7 CT3 40 50 85 100 197 200 14 7 7 CT4 40 90 85 100 157 200 14 7 7 CT5 10 50 115 100 197 200 14 7 7 CT6 10 90 115 100 157 200 14 7 7

37

CT7 40 50 115 100 167 200 14 7 7 CT8 40 90 115 100 127 200 14 7 7 CT9 25 50 100 100 197 200 14 7 7

CT10 25 90 100 100 157 200 14 7 7 CT11 10 70 100 100 192 200 14 7 7 CT12 40 70 100 100 162 200 14 7 7 CT13 25 70 85 100 192 200 14 7 7 CT14 25 70 115 100 162 200 14 7 7 CT15 25 70 100 100 177 200 14 7 7 CT16 25 70 100 100 177 200 14 7 7 CT17 25 70 100 100 177 200 14 7 7

Tin hnh o trn chy, ch s nn v hm m cc khi bt kp ca cc cng thc thc nghim. Kt qu th hin trong bng 17.

Bng 17. Tc chy, ch s nn v hm m ca cc khi bt kp Mu Tc

chy (g/s) Ch s

nn (%) Hm m

(%) Mu Tc

chy (g/s) Ch s

nn (%) Hm m

(%) CT1 13,65 21 2,5 CT9 12,25 22 2,5 CT2 7,14 26 3,2 CT10 8,32 25 3,6 CT3 13,42 21 2,8 CT11 10,59 23 3,0 CT4 6,93 26 3,1 CT12 9,97 24 3,1 CT5 13,87 21 2,6 CT13 9,23 24 3,0 CT6 8,56 25 3,5 CT14 10,74 23 2,8 CT7 12,91 22 2,9 CT15 10,28 23 2,5 CT8 6,98 26 3,8 CT16 10,06 23 2,7

CT17 10,13 23 2,8

Nhn xt: - CP934P c nh hng nhiu nht n tc chy ca khi bt. V d

CT1, CT2 tng khi lng CP934P (50 90 mg), tc chy ca khi bt gim i nhiu (13,65 7,14 g/s), ch s nn tng (21 26%) do CP934P d ht m. Cc CT2, CT4, CT8 c t l CP934P ln (90mg) nn c tc chy thp nht.

38

- Natri hydrocarbonat dng bt kh ri, khng vn cc nn lm tng tc chy ca khi bt nhng khng nhiu. V d CT2, CT6 lng natri hydrocarbonat tng (85 115mg), tc chy ca khi bt tng ln (7,14 8,56 g/s).

- Hm m ca cc khi bt kp dao ng trong khong t 2,5 3,8%. Cc mu vin c bo ch bng phng php dp thng, mi m 100

vin. Kt qu nh gi mt s ch tiu cht lng ca cc mu vin c th hin trong bng 18.

Bng 18. nh gi mt s ch tiu cht lng ca cc mu vin

Mu vin

Ch tiu Mu vin

Ch tiu Hnh thc

bn c hc (kp) (n= 10)

KLTB vin (mg)

(n = 20)

Hnh thc

bn c hc (kp) (n = 10)

KLTB vin (mg)

(n = 20)

CT1 + 11,5 0,8 703,5 6,2 CT9 + 12,3 0,7 701,2 10,1

CT2 + 12,1 1,1 691,4 10,5 CT10 + 11,5 1,2 685,9 7,7

CT3 + 11,2 0,3 695,2 7,8 CT11 + 11,8 0,4 701,5 9,6

CT4 + 12,2 0,9 687,6 10,2 CT12 + 11,9 0,7 689,1 11,4 CT5 + 11,1 0,5 698,5 9,8 CT13 + 12,2 0,6 675,6 10,6

CT6 + 11,7 0,2 695,7 8,1 CT14 + 11,4 0,8 700,5 8,2

CT7 + 12,4 0,4 704,4 6,3 CT15 + 12,4 0,7 696,5 6,7

CT8 + 11,8 1,2 679,7 9,9 CT16 + 11,8 0,5 694,4 8,7 CT17 + 11,7 0,7 698,7 9,4

(Du + biu th vin t yu cu v hnh thc: Vin di, mu trng, bng p, thnh v cnh lnh ln). Nhn xt: Cc mu vin bo ch u t yu cu v hnh thc, bn c

hc (11 13 kp) v ng u khi lng (665 735 mg). Cc CT2, CT4, CT8 tuy c tc chy thp nhng khi dp vin vn t yu cu v ng u khi lng.

Kt qu th kh nng GPDC v KDSH ca cc mu vin bo ch c th hin trong bng 19.

39

Bng 19. Kh nng GPDC v KDSH ca cc mu vin

Nhn xt: Cc mu vin bo ch vi thnh phn CT khc nhau c tc GPDC v lc KDSH khc nhau. Chng t thit k th nghim ph hp, phn nh c nh hng ca cc thnh phn trong CT ti kh nng GPDC v KDSH.

Kt qu nh gi kh nng ni v kh nng trng n ca cc mu vin bo ch c th hin trong bng 20.

Bng 20. Kh nng ni v kh nng trng n ca cc mu vin Mu vin

Kh nng ni Mc trng n theo thi gian (%) tlag

(pht) tni

(gi) 10 pht 20 pht 30 pht 40 pht 50 pht 60 phtCT1 15 >12 21,52 32,44 39,52 47,64 53,64 59,69 CT2 - - 44,85 59,75 72,84 83,25 86,51 94,35

Mu vin

Phn trm ACV gii phng theo thi gian (%) Lc KDSH (N/cm2) 1 gi 2 gi 4 gi 6 gi 8 gi

CT1 28,77 51,35 80,31 90,34 96,4 0,29

CT2 19,7 30,81 50,41 72,76 88,9 0,45

CT3 13,92 27,67 40,26 59,64 73,1 0,32

CT4 16,06 24,56 37,12 52,74 63,3 0,48

CT5 20,11 34,37 57,02 77,6 86,3 0,31

CT6 16,81 28,82 50,51 70,29 82,0 0,46

CT7 11,36 17,41 37,19 50,41 63,9 0,35

CT8 13,84 21,63 35,6 48,92 58,2 0,49

CT9 17,98 32,4 55,35 74,13 88,9 0,33

CT10 15,64 27,72 47,73 67,86 80,4 0,47

CT11 19,08 33,29 56,73 79,88 88,5 0,41

CT12 16,26 30,39 45,33 65,45 80,0 0,44

CT13 19,09 31,92 50,51 79,92 93,6 0,40

CT14 16,94 31,57 47,29 73,85 83,2 0,42

CT15 19,35 32,71 46,48 72,08 89,6 0,41

CT16 18,99 31,53 44,42 70,75 86,1 0,42

CT17 18,64 33,65 47,56 74,8 90,8 0,42

40

CT3 22 >12 27,56 38,79 47,64 54,12 63,22 69,67 CT4 - - 49,41 65,81 79,95 85,62 96,21 100,02 CT5 10 >12 23,95 33,25 41,24 49,34 55,69 62,93 CT6 40 >12 46,26 61,36 75,56 86,83 90,98 96,41 CT7 18 >12 29,78 42,68 53,25 59,92 66,84 72,82 CT8 - - 48,92 67,97 80,61 87,59 95,36 102,62 CT9 17 >12 25,85 35,56 44,96 51,36 58,74 65,43

CT10 43 >12 48,12 63,26 77,54 81,84 89,16 95,56 CT11 28 >12 31,15 45,18 56,20 64,62 71,27 78,48 CT12 35 >12 42,21 57,95 69,55 82,13 85,59 90,31 CT13 30 >12 34,54 48,76 59,36 67,46 75,46 81,57 CT14 25 >12 39,55 55,12 67,87 77,51 81,54 88,37 CT15 27 >12 36,38 51,65 63,56 72,32 78,38 84,16 CT16 25 >12 35,45 50,79 62,55 73,01 77,49 85,67 CT17 29 >12 36,02 52,31 65,42 76,52 79,62 86,43

(Du -biu th vin khng ni) Nhn xt:

V kh nng ni: C ch ni ca vin l do polyme trng n ng thi to ra mt lp gel bao

xung quanh b mt vin gi li kh CO2 to ra bi phn ng gia natri hydrocarbonat vi dung dch acid clohydric do lm gim t trng ca vin. Khi t trng ca vin nh hn t trng ca mi trng th vin s ni trn b mt mi trng. Do vy c CP934P, HPMC K100M v natri hydrocarbonat u nh hng ti kh nng ni ca vin. C th l:

- Khi tng lng natri hydrocarbonat trong CT th thi gian tim tng gim i do to ra nhiu kh CO2 hn nn vin ni nhanh hn. V d CT1, CT5 lng natri hydrocarbonat tng ln (85 115 mg) th thi gian tim tng gim (15 10 pht).

- Khi tng lng polyme trong CT th thi gian tim tng tng ln do tng nht nn to ra lp gel c hn bao xung quanh vin khin mi trng thm vo

41

vin chm hn dn n cn tr phn ng gia HCl v NaHCO3 lm gim to thnh kh CO2. V d CT9, CT10 lng CP934P tng ln (50 90mg) thi gian tim tng di hn ng k (17 43 pht). Cc CT2, CT4, CT8 c t l CP934 ln (90mg) khng ni trong sut thi gian th.

V kh nng trng n: - Mc trng n ca vin tng ln cng vi s tng lng polyme trong

CT. Trong phm vi kho st, tng t l CP934P th MTN tng ln nhiu hn khi tng t l HPMC K100M. V d CT1, CT2 khi lng CP934P tng (50 90mg) th MTN sau 60 pht tng ln nhiu (59,69 94,35%); CT5, CT7 khi lng HPMC K100M tng (10 40 mg) th MTN sau 60 pht tng t 62,93 72,82%. Tuy nhin khi tng lng CP t 50 70mg th MTN tng ln nhiu, v d CT9, CT15, MTN sau 60 pht tng t 65,43 84,16% nhng khi tng lng CP ln na th MTN tng t hn, v d CT15, CT10 lng CP tng 70 90mg MTN sau 60 pht tng t 84,16 95,56%. Do CP nng cao s to ra lp gel c hn lm cn tr tc thm nc t mi trng vo vin nn MTN tng ln khng nhiu.

- Tng t l natri hydrocarbonat trong CT lm MTN ca vin tng ln. V d CT13, CT14 tng khi lng natri hydrocarbonat (85 115mg) th MTN sau 60 pht tng (81,57 88,37%). Do CP934P khi trung ha bng natri hydrocarbonat s c nht cao hn nn kh nng trng n tt hn. 3.3.2. nh gi nh hng ca cc bin c lp ti cc bin ph thuc

S dng phn mm INForm 3.1 x l cc s liu thu c, kt qu c trnh by trong bng 21.

Bng 21. Bng h s tng quan t Y1 n Y8 v KDSH Bin R2train R2test Y1 94,08 86,62 Y2 95,63 91,54 Y4 95,57 92,42 Y6 96,40 90,74 Y8 92,88 89,75

KDSH 99,25 97,72

42

Nhn xt: Gi tr R2train v R2test u cao (> 80%). Nh vy quan h gia cc bin u ra v cc bin u vo l kh cht ch. nh hng ca cc bin c lp ti phn trm gii phng dc cht:

nh hng ca t l Carbopol 934P (hnh 12, hnh 13):

Hnh 12. Mt p nh hng ca Carbopol 934P v natri hydrocarbonat ti Y6 ( Khi lng ca HPMC K100M l 10mg)

Khi tng t l CP934P trong cng thc th tc gii phng dc cht gim i. V CP934P c nht cao, trng n mnh nn to ra lp hng ro gel bao xung quanh b mt vin khin cc knh dn b bt kn nhiu hn, hn ch s thm ca mi trng vo vin dn n ho tan v khuch tn dc cht ra ngoi lu hn. Do khi tng nng CP934P lm tc gii phng dc cht gim i ng k.

nh hng ca t l natri hydrocarbonat (hnh 12): Khi t l natri hydrocarbonat tng ln, kh nng gii phng dc cht gim

xung. Do natri hydrocarbonat trung ha CP934P lm tng nht ca CP934P nn to ra lp gel c hn do lm dc cht khuch tn chm qua hng ro gel hn. Tuy nhin so vi CP934P th mc nh hng ca natri hydrocarbonat t hn.

Y6 (%)

NaHCO3 (mg)

CP 934P (mg)

43

Hnh 13. Mt p nh hng ca Carbopol 934P v HPMC K100M ti Y8 ( Khi lng ca NaHCO3 l 115mg)

nh hng ca t l HPMC K100M (hnh 13): Khi tng t l HPMC K100M trong cng thc vin th tc gii phng

dc cht gim i r rt. V HPMC K100M l polyme c khi lng phn t ln, nht cao, trng n tt v c ch nh hng ca HPMC K100M ti kh nng gii phng ca ACV cng tng t nh CP934P. Trong phm vi kho st c th thy mc nh hng ca HPMC K100M ti kh nng GPDC nhiu hn so vi CP934P. nh hng ca cc bin c lp ti kh nng KDSH:

nh hng ca Carbopol 934P n lc KDSH (Hnh 14, hnh 15):

Y8 (%)

Lc KDSH (N/cm2)

CP 934P (mg) NaHCO3

(mg)

Hnh 14. Mt p nh hng ca Carbopol 934P v natri hydrocarbonat ti lc KDSH (Khi lng ca HPMC K100M l 25 mg)

CP 934P (mg)

HPMC K100M (mg)

44

Mt p hnh 14 v 15 u cho thy trong phm vi kho st, lng CP934P c nh hng ch yu ti kh nng KDSH. Tng t l CP934P trong CT th kh nng KDSH tng ln r rt. Khi CP934P tng t 50 70mg lc KDSH tng ln nhiu, CP934P tng t 70 90mg lc KDSH tng ln nhng t hn. iu ny c gii thch l do khi CP nng thp, tng t l CP lm tng nht to ra lp gel c hn nn lc KDSH tng ln nhiu. Tuy nhin khi tng nng CP n mc no th lp gel bo ha do c tng thm CP ln na cng khng thay i nhiu th cht ca lp gel nn lc KDSH tng khng ng k.

nh hng ca natri hydrocarbonat n lc KDSH (Hnh 14): Khi tng lng natri hydrocarbonat trong cng thc, lc KDSH tng ln. Do

natri hydrocarbonat trung ha CP934P lm tng nht ca CP934P nn lm tng kh nng KDSH. Tuy nhin so vi CP934P th mc nh hng ca natri hydrocarbonat ti kh nng KDSH khng ng k.

nh hng ca HPMC K100M n lc KDSH (hnh 15):

Khi lng HPMC K100M trong vin tng lc KDSH cng tng nhng mc

nh hng khng ng k so vi CP934P. V HPMC K100M c nht cao nn cng phi hp vi CP934P lm tng kh nng kt dnh trn nim mc.

Tm li, t mt p hnh 14, hnh 15 c th thy rng kh nng KDSH ca vin ch yu c quyt nh bi t l CP934P trong cng thc.

Lc KDSH (N/cm2)

HPMC K100M (mg)

CP934P (mg)

Hnh 15. Mt p nh hng ca Carbopol 934P v HPMC K100M ti lc KDSH (Khi lng ca NaHCO3 l 100mg)

45

3.3.3. Ti u ha cng thc vin nn acyclovir kt dnh sinh hc ng tiu ha Ti u ha cng thc bng phn mm INForm 3.1 theo yu cu bin u ra

th hin trong bng 15. Cng thc vin nn ACV KDSH c d on nh sau: Acyclovir 200,00 mg Carbopol 934P 71,10 mg HPMC K100M 10,00 mg Natri hydrocarbonat 98,46 mg Lactose 100,00 mg Avicel PH101 192,44 mg Magnesi stearat 14,00 mg Talc 7,00 mg Aerosil 7,00 mg Tin hnh bo ch 3 m vin nn, mi m 100 vin theo cng thc ti u sau

nh gi kh nng GPDC v KDSH. Kt qu c trnh by trong bng 22 v hnh 16.

Bng 22. Kh nng GPDC v KDSH ca mu vin bo ch theo cng thc ti u v d on

Mu vin Phn trm ACV gii phng (%) f2 Lc KDSH (N/cm2) 1 gi 2 gi 4 gi 6 gi 8 gi Vin bo ch

theo CT ti u 19,78 0,45

35,78 1,45

56,77 1,3

70,2 0,51

88,63 2,04

83,9 0,41

0,05 D on 18,05 36,07 55,15 68,78 91,67 0,41

Hnh 16. th GPDC ca vin bo ch theo cng thc ti u v d on

0102030405060708090

100

0 2 4 6 8 10

% A

CV

gii

ph

ng

Thi gian (gi)

CT ti uCT d on

kho

khAC

tron

u

H

Nhnong 50 1 nng GPD

CV. Do , n

c kt qu:ng qu trnh

c kh nn

Hnh 17. H

c

A

n xt: T c100). Nh vDC ko di

m hnh thi

nh gi kh : Thi gianh ni khng

ng ni ph h

Hnh nh viclohydric 0,

cc kt quvy, vin nging vi mit k th ngnng ni c

n tim tng g c hin thp vi m

n bo ch t,1M (A: 0 g

B

trn, ch sn ACV KD

m hnh dghim la mu vil 25 2,8

ng r. Chhnh GPDC

theo cng th

gi; B: Sau

46

s f2 c giDSH bo ch

on, sau a chn l pn bo ch82 pht, thhng t muC ko di 12

hc ti u n30 pht; C

C

tr bng 8h theo cng8 gi gii p

ph hp. theo cng

i gian niu vin bo

2 gi.

ni trong duC:Sau 12 gi

83,9 (nm tg thc ti phng trn

thc ti ui hn 12 gich theo CT

ung dch aci)

trong

u c 80%

u thu

i v T ti

cid

47

PHN 4. KT LUN V XUT 4.1. Kt lun

Sau qu trnh tin hnh lm thc nghim, chng ti thu c mt s kt qu chnh nh sau:

La chn phng php dp thng bo ch vin nn acyclovir KDSH. La chn polyme xy dng cng thc bo ch vin nn acyclovir

KDSH, l hn hp 2 polyme CP934P v HPMC K100M. La chn t dc to kh l natri hydrocarbonat gip cho vin ni. nh gi nh hng ca cc t dc: CP934P, HPMC K100M v natri

hydrocarbonat n mt s ch tiu cht lng ca vin ACV KDSH: Kh nng gii phng dc cht: T l CP934P v HPMC K100M tng ln th tc GPDC gim i r rt. T l natri hydrocarbonat tng th tc GPDC cng gim nhng mc

nh hng t hn so vi CP934P v HPMC K100M. Kh nng kt dnh sinh hc in vitro: CP934P ng vai tr quan trng v quyt nh ti kh nng KDSH ca

vin thuc. T l CP934P tng th kh nng KDSH tng ln r rt. Tng t l HPMC K100M v natri hydrocarbonat trong CT th lc KDSH

tng ln nhng mc nh hng t hn so vi CP934P. Kh nng trng n: Tng t l CP934P, HPMC K100M, natri

hydrocarbonat th mc trng n ca vin tng ln trong CP934P c nh hng r rt nht.

Kh nng ni in vitro: T l natri hydrocarbonat tng th thi gian tim tng gim i. T l CP934P v HPMC K100M tng th thi gian tim tng tng ln. Xy dng c cng thc bo ch vin nn acyclovir ni kt dnh sinh

hc ng tiu ha gii phng ko di 12 gi: Acyclovir 200,00 mg

Carbopol 934P 71,10 mg

48

HPMC K100M 10,00 mg

Natri hydrocarbonat 98,46 mg

Lactose 100,00 mg

Avicel PH101 192,44 mg

Magnesi stearat 14,00 mg

Talc 7,00 mg

Aerosil 7,00 mg

4.2. xut tip tc pht trin dng bo ch ny, chng ti a ra mt s xut nh sau:

Nghin cu n nh ca ch phm. Nghin cu bo ch vin nn acyclovir KDSH qui m ln hn c c s ng dng trong thc t sn xut. Tin hnh th nghim in vivo khng nh kt qu th nghim.

TI LIU THAM KHO Ting Vit 1. B Y t (2007), Dc l tp 2, NXB Y hc, H Ni, tr. 233 235. 2. B Y t (2007), Ha dc tp 2, NXB Y hc, H Ni, tr. 231 233. 3. B Y t (2008), K thut bo ch v sinh dc hc cc dng thuc tp 2, NXB

Y hc, H Ni, tr. 61 63. 4. B Y t (2009), Dc in Vit Nam IV, NXB Y hc, H Ni. 5. B Y t (2009), Dc th quc gia Vit Nam, tr. 102 104. 6. Don Th Thu Hin (2009), Nghin cu bo ch vin nn acyclovir kt dnh sinh

hc t ph khoa, Lun vn thc s dc hc, Trng i hc Dc H Ni. 7. Nguyn Thu Hin (2011), Nghin cu bo ch vi cu acyclovir kt dnh sinh hc

gii phng ko di, Kha lun tt nghip Dc s i hc, Trng i hc Dc H Ni.

8. Trn Th Thy Nga (2009), Nghin cu bo ch vin nn acyclovir kt dnh sinh hc t ph khoa, Kha lun tt nghip Dc s i hc, Trng i hc Dc H Ni.

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