HNG NGHIN CU

Nghin cu Vt l trong ha hc (khoa hc trong khoa hc) S khi l c trng c bn nht ca nguyn t hp cht hu c T hp C, H, N, O, S...

Hng nghin cu

Nghin cu ha hc trong ha hc

Nghin cu ha hc trong y hc (khoa hc trong khoa hc s sng) - Pht hin v pht trin thuc mi

S khi chnh xc ca mt s nguyn t ca hp cht hu c Nguyn t Trng lng Ng.t ng v

1H 2H

S khi

Hydro

1,00797

1,00783
2,01410 12,0000 13,00336 14,0031 15,0001 15,9949 16,9991 17,9992 30,9738 31,9721 32,9715 33,9679 34,9689 36,9659 78,9183 80,9163 126,9045

Carbon Nit

12,01115

12C 13C

14,0067

14N 15N

Oxy

15,9994

16O 17O 18O

Phosphor Lu hunh

30,974 32,064

31P 32S 33S 34S

Clo

35,453

35Cl 37Cl

Brom

79,909

79Br 81Br

Iod

126,904

127I

S pht trin ca cc phng php vt l trong cng ngh thit b t ti trnh rt cao, o chnh xc nng lng, s khivv S pht trin mnh m ca MS Ci tin v nhiu cng ngh mi trong:
K thut ion ha (ionisation) v phn mnh (fragmentation) K thut pht hin s khi (detection, mass analyzer)

MT S K THUT PHN TCH KHI PH HIN I TRONG PHT HIN V PHT TRIN THUC

Sau khong 100 nm hnh thnh, MS pht trin rt mnh m Nhiu th mnh trong NCKH, c bit l ha hc thuc. Cc thit b MS c th s b lit k t k thut ion ha khc nhau: Va chm in t EI, bn ph nguyn t nhanh FAB, ion ha bng ha hc CI, ghp ni GC MS Phun in t ESI, ion ha p sut thng APCI, Ghp ni tandem LC MS/MS, MALDI, TOF (Time of Flight), Phn tch ICP - MS...vv. c bit k thut phn tch ph khi phn gii cao v chnh xc khi cao da trn nguyn l chuyn ha Fourier cng hng ion cyclotron (FT ICR MS) Thit b phn tch ph khi c chnh xc khi cao (Accurate MS) trn nguyn l by orbital (orbitrap) l nhng ng dng rt c tim nng trong pht trin thuc mi.

Mt s h thng thit b khi ph (MS) v sc k lng khi ph (LC MS) dng trong nghin cu pht hin v pht trin thuc

2004
LC MS/MS (Ion trap) Thermo Finnigan LCQ Advantage Max

H thng LC MS by ion Iontrap LCQ Advantage Max, Khoa Vt l o lng, Vin KN Thuc Trung ng

Mt s h thng thit b khi ph (MS) v sc k lng khi ph (LC MS) dng trong nghin cu pht hin v pht trin thuc

2009
H thng LC MS 3 t cc ni tip Thermo Finnigan TSQ Quantum Ultra, Trung tm Tng ng sinh hc,Vin KN Thuc Trung ng LC MS/MS (Triple Quadrupoles)

Thermo Finnigan TSQ Quantum Ultra

Mt s h thng thit b khi ph (MS) v sc k lng khi ph (LC MS) dng trong nghin cu pht hin v pht trin thuc

2009

H thng ph khi chnh xc cao Thermo LTQ XL (Orbitrap), Phng th nghim trng im ha Ha dc, H KHTN, i hc Quc gia

Mt s h thng thit b khi ph (MS) v sc k lng khi ph (LC MS) dng trong nghin cu pht hin v pht trin thuc

2009

H thng ph khi phn gii cao Varian 900-MS Series FTMS, Vin Ha hc, Vin KHCN Vit Nam

FT ICR MS siu phn gii khi, o chnh xc s khi

Glucagon l 1 hormon tuyn ty, c bn cht l 1 poly-peptid

Hng nghin cu trong y hc

Nghin cu nng cao (mang tnh hn lm, Acadmic researches)

Pht hin thuc mi & pht trin thuc mi Chng minh c ch, nguyn l, lin quan cu trc tc dng sinh hc...vv

Nghin cu thng dng

Pht trin thuc mi (industrial R&D labo...) ng k lu hnh thuc Qun l cht gim st cht lng thuc ...vv.

Mt s nghin cu thng dng tr thnh thng quy Trong lnh vc kim nghim, gim st cht lng thuc v an ton thc phm, MS kt ni sc k (GC MS, LC MS/MS), c bit LC-MS tr thnh phng php thng quy trong:

Chng thuc gi v gian ln thng mi (tn dc trong ng dc); D lng khng sinh (chloramphenicol, streptomycin)...trong thy sn; Phn tch c t nm aflatoxin trong thc phm, dc liu; Phn tch melamin trong sa v cc sn phm t sa; D lng cht tng trng clenbuterol, salbutamol trong tht gia cm, D lng cht dit c 2,4 D, 2,4,5 T, Ha cht BVTV (c clo, c phosphor, pyrethorid) trong dc liu, sn phm t dc liu ...; Phn tch thuc trong dch sinh hc trong qu trnh pht trin thuc mi, trong nh gi hoc theo di lm sng... Trong nghin cu c cht php y: phn tch cc cht ma ty tng hp nhm amphetamin (ATS), nhm gy nghin Narcotic; Phn tch cc cht doping trong th thao; Phn tch cht c chin tranh dioxinvv

QU PHN DOANH S TI H NI THNG 5 TH TRNH PHT TRIN MT THUC MI

PHT HIN (discovery) PHT TRIN THUC (develop.)

NC Tin lm sng
CU TRC LM THUC

3 5 nm 3 nm

5.000 hp cht

thuc

PHT TRIN THUC (develop.)

Phase 1

Phase 2

Phase 3

15 17 nm FDA

NC, nh gi lm sng

3 nm 6 nm

Pht hin thuc mi (3-5 nm).

Qu trnh pht hin thuc mi bt u vi vic sng lc hot tnh sinh hc (hip ng, i khng hay cnh tranh) ca cc hp cht khc nhau t ngun tng hp ha hc hoc t cc hp cht thin nhin trn th th ch nh cc receptor mng t bo lin kt protein G hoc protein kinase. T l thnh cng ca giai on ny thng khong 0,1%, c 5.000 ngn cht may ra sng lc c 5 cht c th lm thuc.

nh gi tin lm sng (khong 3 nm)

giai on ny, cc hng dc phm tin hnh cc th nghim phng th nghim v nghin cu trn ng vt th nghim th hot tnh sinh hc ca hp cht i vi bnh. Hp cht cng c nh gi v s an ton.

Pht trin thuc nh gi lm sng

Thuc th nghim, IND. i vi thuc mi ang thm d cn gi l thuc th nghim (The Investigational New Drug, gi l thuc IND), phi ng k vi vi Cc Dc Thc phm (Food and Drug Administration, FDA) sau khi c nh gi v th tin lm sng. Thuc IND m tha mn cc yu cu trn, th lc , s phi th nghim trn ngi. Thuc IND bao gm cc thng tin v cu trc ha hc, tc dng trn ngi nh th no, c tnh ra lm sao v phng php sn xut thuc. Th lm sng, Phase I. Th nghim trn 20 80 ngi tnh nguyn khe mnh kim tra tnh an ton, thit lp liu an ton v thi gian tc dng. Cc nghin cu tp trung vo qu trnh hp thu, phn b, chuyn ha v thi tr ca thuc (cc nghin cu ADME). Th lm sng, Phase II. Thuc c th trn 100 300 bnh nhn tnh nguyn m ang mc loi bnh cn th. Hiu lc iu tr ca thuc s c nh gi. Th lm sng, Phase III. Thuc c tip tc th nghim trn 1.000 3.000 bnh nhn, cc bc s s kim tra hiu qu cng nh tc dng ph ca n.

Pht trin thuc

ng k thuc mi, NDA. Cng ty phn tch cc d liu nghin cu, v nu nh thuc an ton v hiu qu, cng ty s lm h s ng k thuc mi (New Drug Application, NDA) vi FDA. FDA s yu cu t nht 6 thng nh gi, thm nh h s thuc mi, bao gm tt c cc d liu nghin cu bo co. Chp thun. Thuc c s dng v c k n. FDA s yu cu nh k bo co v thuc, v cn phi c cnh bo nu nh c bt c cc phn ng ph no ca thuc.

NG DNG CA PHN TCH PH KHI TRONG PHT HIN V PHT TRIN THUC MI

Trong pht hin thuc (discovery)

Tng hp ha hc Sng lc thin nhin

Trong pht trin thuc (development)

Sng lc cc ligand, thit k cc phn t lm thuc da vo tng tc vi th th ch Protein G, Nobel y hc 1994

Pht hin thuc

Alfred Goodman Gilman (sinh ngy 1.7.1941) l mt nh dc l hc, nh ha sinh ngi M, ot gii Nobel Sinh l v Y khoa nm 1994 chung vi Martin Rodbell cho cng trnh khm ph ca h v cc protein G. Protein G l mt cht trung gian cn cho s sng gia s hot ha ngoi t bo ca cc c quan nhn G protein-coupled receptor (GPCR) mng t bo v cc hot ng bn trong t bo.

Sng lc cc ligand, thit k cc phn t lm thuc da vo tng tc vi th th ch

Lin kt gia phn t thuc (ligand) Protein ch (target) c phn t lng ln

Lin kt = khng cng ha tr (noncovalent) lin kt i lc (affinity) lin kt c hiu (specifity)

Sng lc cc ligand, thit k cc phn t lm thuc da vo tng tc vi th th ch

Agonist = c tc dng

Antagonist = lm ko c tc dng

Sng lc cc ligand, thit k cc phn t lm thuc da vo tng tc vi th th ch

Hp cht d kin (candidate): hp cht thin nhin, tng hp Hp cht kim tra cht lng Tnh ha tan Cng thc bo ch Khung (panel) sng lc candidate + ch (target) kim tra cht lng ch p.t ch ADN, ARN, protein i lc NC ha Kd hc Tnh c hiu trong y T l hp thc ha hc hc cht khi u lm thuc (Leads) c trng ha cu trc MS/MS, IRMPD-MS, IMS Gia cng sa cha ha hc (refined hits) i lc vi ch

NC ha hc trong y hc

Hng mi trong nghin cu pht hin thuc mi, rt c tim nng Smith v cng s (2001)

MASS

Gia cng sa cha li khung (refined panel)

DOLCE

SAR by MS
Hp cht p ng yu cu (hits)

NC ha hc trong y hc

Pht hin thuc

Sng lc th cc hp cht thin nhin

T l thnh cng, t chi ph hn so vi t ngun tng hp Li ch khi ng dng FT ICR MS (cc c tnh v hiu nng)

chnh xc khi cao xc nh cng thc phn t phn gii khi rt cao chia tch cc thnh phn, phn tch ng thi Kh nng thc hin cc mc phn tch khi nhiu ln lm sng t cu trc nhy cao phn tch c cht phn lp thng hn ch v lng

C rt nhiu ng dng s dng cc tnh nng ny ca FT ICR MS c cng b gn y v cu trc cc hp cht thin nhin c hot tnh sinh hc t cy thuc v t sinh vt bin. Hnh 5 l mt v d v ng dng FT ICR MS trong d on cng thc v cu trc ca methylspirastrellolide A sng lc t vi sinh vt sinh tng hp thuc mi.

Giai on pht trin thuc mi (drug development)sng trong c Giai on ny, vic nghin cu dc l, tin lm

cc nghin cu dc ng hc ca thuc mi th nghim IND (hp thu, phn b, chuyn ha, thi trNghin cu ADME). Vic nh lng thuc v xc nh cht cht chuyn ha ca thuc trong cc dch sinh hc ca c th l cn thit. Trong giai on ny, cc cht thng bit v cu trc cng nh c ch tc dng, nhng sn phm chuyn ha ca n th thng cha c bit. Cc h thng khi ph ghp ni sc k kh (GC EI MS) hoc sc k lng c nhy cao (LC ESI MS) thng hay c s dng nht nh lng hot cht trong mu, huyt thng v cc dch th sinh hc khc. Do tnh tng thch vi phn tch dch sinh hc, LC - MS ngy cng c s dng rng ri v chim u th hn GC - MS (ch thch hp phn tch cc hp cht d bay hi). a s cc phng php phn tch thuc trong dch sinh hc thng s dng LC MS/MS, trong h sc k c th l sc k lng hiu nng cao (HPLC), sc k phn gii nhanh (RRLC) hoc sc k siu hiu nng (UPLC) v detector khi ph thng l loi 3 t cc ghp ni tip (triple quadrupoles).

Nghin cu tng ng sinh hc BE ca thuc gc

Thuc pht minh
Patented drug, Invented Drug Thuc gc Generic drug Super generic drug (TQ, n , Brazin)

Oseltamvir Bn quyn Roche 27/12/1996 (U.S. Patents) 20 nm, gi tr n ht 26/12/2016.

Taxol Bristol-Myers Squibb, 03/03/1992 (U.S. Patents) 02/03/2012

Bn quyn Pfizer 1996, Pfizer cng b bt u nhng quyn i tr ton th gii 2011 - 2013

Viagra Sildenafil

ng cong DH ca 14 NTN

Nng (ng/mL) 700

600

500

400

300

200

100

0 -1

14

19

24

Thi gian (gi)

KT QU NGHIN CU
ng cong DH NTN14
Nng (ng/mL)
700 600 500 400 300 200 100 0 0 10 20 30 40 50 60 70 80 90
ng ng cong dc ng hc NTN 14 cong dc ng hc NTN 14 700 Azithromycin Zitromax

Azithromycin Zitromax

Nng (ng/mL)

600 500 400 300 200 100 0 0 10

20

30 Thi gian (gi)

Thi gian (gi)

NC nh lng olanzepin trong huyt tng ngi

H:\C4 RT : 0.00 - 3.25 100 50 0 100 50 0 100 50 0 0 1 2.27 2 T ime (min) 3 0.12 1.36 2.14 2.64 0.12 1.36 1.68
NL: 7.68E4 TIC F: + c ESI SRM ms2 31 3.000 [255.750-256.250] M S C4 NL: 1 .73E5 TIC F: + c ESI SRM ms2 31 9.000 [85.750-86.250] M S C4

2.64 1.68

NL: 1 .73E5 TIC M S C4

Sc k mu huyt tng cha OLZ v CLZ nng tng ng 1 ng/mL v 10 ng/mL

KT LUN
Trong vi nm li y, cc thit b khi ph hin i c trang b mt s c s nghin cu nh h thng MS phn gii v chnh xc cao nh Thermo Finnigan LTQ XL (Phng th nghim trng im Ha dc, Trng H KHTN, i hc Quc gia H Ni), h thng sc k lng siu hiu nng ghp ni khi ph 3 ln t cc ni tip (LC MS/MS) Thermo Finnigan TSQ Quantum Ultra (Vin Kim nghim Thuc trung) v c bit l h thng khi ph phn gii cao v chnh xc cao FT ICR MS (Vin Ha hc, Vin KH&CN Vit Nam)... v ang bt u c khai thc s dng trong nghin cu KH. Vi mt s ng dng nh nu, chng ti mong mun v hy vng c s hp tc v khai thc c hiu hiu qu cc thit b trn trong nghin cu khoa hc c bit l trong lnh vc pht hin v pht trin thuc mi, gp phn vo s nghim chm sc v bo v sc khe nhn dn.

Xin chn thnh cm n! The end

The end