B Y T TRNG I HC DC H NI BCH C HIPHA HC CA CC CHT C CH DIPEPTIDYL PEPTIDASE-4 DNG TRONG IU TR TIU NG KHA LUN TT NGHIP DC S H NI - 2013B Y T TRNG I HC DC H NI BCH C HIPHA HC CA CC CHT C CH DIPEPTIDYL PEPTIDASE-4 DNG TRONG IU TR TIU NG KHA LUN TT NGHIP DC S Ngi hng dn: PGS.TS. Nguyn Hi NamNi thc hin:B mn Ha Dc H NI - 2013LI CM NNgi thy dnh thi gian v tm huyt hng dn tn tnh, ch bo cho ti nhiu kin thc qu bu trong sut qu tnh thc hin ti.Ti xin gi li cm n n, DS. Hong Vn Hi cng cc thy c gio trong b mn Ha dc ging dy v to iu kin thun li cho ti trong sut qu trnh hc tp v hon thnh kha lun.Ti cng xin gi li cm n cc thy c trong Ban gim hiu, Phng o to v cc thy cgiotrngihcDcHNidydvgip ti trong sut qu trnh hc tp ti trng.Cuicngtixinbytlngbitnsusctigianh,bnbcngnhnhngngi thn yu lun chia s, ng vin, gip ti trong hc tp v trong cuc sng.H Ni, ngy 16 thng 5 nm 2013Bch c HipDANH MC CC K HIU, CH VIT TTT2DM = Type 2 Diabetes T2DM = Type 2 Diabetes Mellitus DPP= Dipeptidyl peptidaseGLP-1 = Glucagon-like peptide-1GIP = Gastric inhibitory peptideHbA[1c] = Glycosylated hemoglobin A1cFDA = US Food and Drug AdministrationOGTT = Oral glucose tolerance testTZDs = ThiazolidinedionesSUs = SulphonylureasSAR = Structure-activity relationshipsT = i tho ngPOP= Prolyl oligopeptidase HDL = Lipoprotein t trng caoLDL = Lipoprotein t trng thpHTS = High-throughput screeningFAP= Fibroblast activation protein DANH MC BNGBng 3.1 39Bng 3.2 51Bng 3.3 52Bng 3.4 52Bng 3.5 52DANH MC CC HNH V, THHnh 1.1: Cc sulfonylurea th h 1 ........................................................................5Hnh1.2: Cc sulfonylurea th h 2.......................................................................5Hnh1.3: Mt s thuc i din ca nhm biguanid...............................................5Hnh1.4: Mt s thuc i din ca nhm thiazolidindion.....................................6Hnh1.5: Mt s i din ca nhm meglitinid [30, 31] ........................................6Hnh1.6: Mt s i din ca nhm.......................................................................7Hnh2.7: c im v tr ca 6 thnh vin trong h DPP-IV,vi sDPP-IV l dng ha tan ca DPP-IV [10] ..........................................................................................................10Hnh2.8: Chui theo hng ngang cc acid amin trn DPP-IV v POP................. 11Hnh2.9: Ton th cu trc ca DPP-IV dng homodimer...................................12Hnh2.10: Cu trc cc monomer [51]................................................................ 14Hnh2.11: Cch tn cng vo DPP-IV.................................................................15Hnh2.12: Cc v tr hot ng trn DPP-IV........................................................ 17Hnh2.13: Diprotin A (IIe-Pro-Ile) v cc v tr hot ng ca DPP-IV.............. 18Hnh2.14: ng vo v tr hot ng ca DPP-IV.............................................19Hnh2.15: V tr cc aa trn GLP-1 v GIP [69].................................................. 20Hnh2.16: V tr tc dng ca DPP-IV.................................................................20Hnh2.17: C ch hot ng ca GLP-1 trong vic iu tit glucose. ................. 21Hnh2.18: Hai cch bo v dng hot ng ca hormon incretin trc tc dng ca enzym DPP-IV [65] ..........................................................................................................21Hnh2.19: Gin c cht ca DPP-IV............................................................... 23Hnh2.20: Tnh chn lc trn DPP-IV ca mt s cht c ch. ............................ 24Hnh3.21: Cc cht dn ng trong tng hp ca DPP-IV. ............................... 26Hnh3.22: Gin c cht ca DPP-IV............................................................... 26Hnh3.23: Cc cht c ch nhm peptid v khng cng ha tr ........................... 27Hnh3.24: Cc cht c ch cng ha tr. ............................................................. 28Hnh3.25: Cc cht dn ng ca cyanopyrrolidin. ..........................................29Hnh3.26: nhm ................................................................................................ 36Hnh3.27: nhm thun nghch v khng thun nghch ......................................37Hnh3.28: Sitagliptin ........................................................................................... 37Hnh3.29: Mt s i din nhm khng gi peptid............................................... 37Hnh3.30:CsthitkcutrcCdavoAlogliptin(A)vccchtcchDPP-IV(B).............................................................................................................................. 42Hnh3.31: Nhng v tr tn cng ca C vi DPP-IV.............................................44T VN Bnhithongtyp2(T2DM)lbnhphbin trn ton th gii v chim gn 6% tngdns.Gn200triungibbnhtiungthcn90-95%sngibT2DM. Theo c tnh s c khong 366 triu ngi b nh hng bi T2DM trong vng 30 nm ti nu cc bin php phng nga khng cn hiu qu [97]. Theo hng thng tin qung co Med s lo havbophcadnsngycngtngsthcythtrngthucT2DMlntikhong 20.5tUSDvonm2012.CcthuciutrngunghinnayiviT2DMbaogm metformin,sulfonylurea(SU),thiazolidinedion(TZD)vccchtcchglycosidase.Chng u gy ra cc tc dng ph nh: gy c ng tiu ha, tng cn, ph n...v do xut hin nhucuphttrinccthucchngithong(T).Trongnhiunmgny,ccliu php iu tr lm tng mc insulin c chng minh l c li cho iu tr T2DM. Cc cht c trng lng phn t thp c ch thun nghch dipeptidyl peptidase 4 (DPP-IV) c nghin cu trong nhiu nm [142] cho thy tc dng h ng huyt tt, tng dung np glucose v ci thin p ng insulin chng li nhng bnh nhn c T2DM [119]. V vy DPP-IV c xem l tim nng cho mc tiu iu tr ca T2DM. Bng chng chng minh hiu qu ca cc cht c ch DPP-IV c chp nhn trn th nghim lm sng ngi v cho thy vic ci thin tt lng glucose [92, 102].DPP-IV v cc cht c ch DPP-IV hin ang c nhiu nh khoa hc trn th gii quan tm,nghin cu. Tuy nhin, Vit Nam, vn ny cn kh mi l. V vy chng ti thc hintiHAHCCACCCHTCCHDPP-4DNGTRONGIUTR TIU NG vi mong mun h thng ha cc cht c ch DPP-IV. Mc tiu ca ti ny l: Trnh by c c im ca dipeptidyl peptidase - 4 v ng dng trong nghin cu pht trin thuc iu tr tiu ng.Trnh by c c s ha hc ca mt s cht c ch peptidyl peptidase- 4 hng dng lm thuc iu tr tiu ng c nghin cu pht trin gn y. PHN 1 : BNH TIU NG V THUC IU TR1.1. Gii thiu v bnh T2DM :1.1.1. nh ngha :Theo t chc y t th gii (WHO), T l mt hi chng c c tnh biu hin bng tng glucose huyt do hu qu ca vic thiu/hoc mt hon ton insulin hoc c lin quan n vic suy yu trong hot ng ca insulin. 1.1.2. Phn loi : Hip hi T M 2009, T c chia lm 4 loi T typ 1; T typ 2; T thai k v cc typ c bit khc. Trong ch yu nghin cu 2 dng l typ1 v typ 2.T typ 1 T typ 1 c trng bi tnh trng thiu insulin tuyt i, hu qu ca s ph hy t bo donguynnhntmin,dovirus,dohachthoctpht(vcn)trongnguynnhn thng gp nht l do t min. Triu chng ca T type 1 bao gm : kht nhiu, n nhiu, i nhiuvgimcn.Tuikhibnhthnggpnhtldi30,tuyvycngcthgpla tui cao hn. Bnh thng c yu t di truyn hoc lin quan n 1 s yu t mi trng (nhim virustrongthikcthai,ct..)vcthmccngvimtsbnhtminkhcnh: basedow, vim tuyn gip Hashimoto, bnh Addision.. T typ 2 : T typ 2 lin quan n s thiu insulin tng i v hin tng khng insulin. Nng insulinmubnhthnghoccaotrongtrnghpkhnginsulinchimuth.Nng insulin gim trong trng hp c khim khuyt chc nng tit insulin. Nh vy,mc d t bo o ty vn tit insulin nhng khng vt qua mc khng insulin do glucose huyt vn tng.So snh s khc nhau gia typ 1 v typ 2 : Typ 1 : Haycngilbnhtmin,tuyntybtncngvphhybichnhcctbot min, do lm bt hot kh nng tit insulin. Nhng ngi b i tho ng typ 1 phi c tim insulin hng ngy.Typ2 : Tuyn ty sn xut insulin, ti thiu trong cc ba n hng ngy ca ngi b bnh tiu ng. Nhng do mt vi nguyn nhn m vn cha c xc nh r, cc t bo ca c th khng li hoc v tnh khng li insulin. b p li, cc bm ca tuyn ty tng cng lng insulin a mc glucose v bnh thng. V theo thi gian, khong 10 nm, vic tng insulin khng c duy tr, v kh nng sn xut insulin ca tuyn ty b suy gim [14] .Hng iu tr :Ngynay,iutrithongchiuqucncsnggpcanhiuchuyn ngnh, nhiu chuyn khoa. iu tr i tho ng cn phi c [1, 2, 136] : Ch dinh dng hp l, tc l phng php iu tr bng ch dinh dngRn luyn c thChng trnh hun luyn bnh nhnThuc lm gim ng khi cn thitCc nhm thuc iu tr tiu ng : iu tr tiu ng c 2 loi chnh : insulin (hormon tuyn ty) v cc thuc tng hp iu tr T.1.2.1. Insulin : Lnutin,insulincchittchthnhcngttuyntycachvonm1922bi Banting v Best. Nm 1951, Sanger v cng s xc nh c cu trc ha hc ca insulin v nm1963,insulinctnghptonphn.Nm1972,cutrckhnggian3chiuca insulin c xc nh bi Hodg-kin cng cng s bng phn tch tia X ca cc n tinh th.Insulin c tng hp cc t bo ca tuyn ty t preproinsulin, mt polypeptid mch n gm 110 aa. Sau , n b tch mt 24 aa cui cng to thnh proinsulin.Insulin c iu ch bng cch phn lp t tuyn ty ca ln, b. Mun iu ch insulin ngi, bn tng hp t insulin ln hoc bng cng ngh ti t hp ADN.Cc thuc tng hp iu tr bnh TThuctnghpchngtngnghuythocthuclmhnghuytctcdngtt trong iu tr bnh i tho ng typ 2. Theo c ch tc dng, c th chia cc thuc ny thnh cc nhm c bn sau : Nhm 1 : sulfonylurea: Nm1942Janbonvngnghipphthinrarng,mtssulfonylureactcdngh nghuyt.Khmphnydnnvica1-butyl-3sulfonylurea(carbutamid)voiutr bnhithong.Vsau,doctnhtrntyxngnnhpchtnykhngdngna. Nm 1950, cht u tin c s dng rng ri trong bnh iu tr i tho ng l tolbutamid, v t , 20 hp cht thuc nhm ny c a vo s dng trong iu tr [73].Tt c cc hp cht nyu l dn cht ca sulfonylurea, khc nhau v tr R1 v R2. Do ctnhacidyunncchpchtnhmnylinktmnhviproteintrongmu.Vvy,khi dng ng thi vi cc thuc khc, chng c th y cc thuc ra khi ch lin kt protein, ckhigytcdngkhngmongmun.Vd,khidngtolbutamidng thi cng dicumarol, c th gy chy mu.Cc sulfonylurea c chia lm 2 th h, th h 2 tc dng mnh v thng lu hn th h 1. Sau y l cng thc cu to ca mt s hp chtTh h 1 (Hnh 1.1) : Hnh 1.1: Cc sulfonylurea th h 1Th h 2 (Hnh 1.2) : Hnh1.2: Cc sulfonylurea th h 2Nhm 2 : biguanid v cc dn cht Thi trung c, chu u, ngi ta dng cy Galera officinalis iu tr bnh i tho ng.uthk20,ccnhkhoahcxcnhchotchttrongcyctcdngl guanadin.Tuyctnhchtlmhnghuyt,nhngctnhcao nn guanadin khng c dng trong iu tr. Mt thi gian ngn sau khi a cc sulfonylurea vo iu tr, mt biguanide utinlphenformincdngtronglmsng.Phenformingynhimacidlacticnnngy nay khng c tip tc s dng. Metformin, mt biguanid khc c tc dng tt, t tc dng ph c s dng rng ri nh hin nay.Mt s thuc i din (Hnh 1.3) : Hnh1.3: Mt s thuc i din ca nhm biguanidNhm 3 : nhm thiazolidindion v cc dn cht : Nm1997,triglitazon,mtTZDutincngcsdngtiM[45].ylmt nhmthucmictcdngchngtngnghuyt.Trongnhmny,hinc2chtc dng trong iu tr i tho ng typ 2 l pioglitazon v rosiglitazon.Hnh1.4: Mt s thuc i din ca nhm thiazolidindionNhm 4 : cc cht tng t meglitinid Ccglinidcng c xem l mt i din mi trong nhm iu tr T2DM,chng c cu trctngtvimeglitinid.Ccglinidkchthchbititinsulin,tcdngnhanhvthigian bn thi ngn. Hot ng bng cch kha knh KATP.Hnh1.5: Mt s i din ca nhm meglitinid [30, 31]Nhm 5 : cc cht c ch -glucosidase Trong nhm ny c 2 cht hay c dng trong iu tr bnh i tho ng l acarbose v miglitol [45]. C ch tc dng chung l c ch thun nghch cc enzym -glucosidase rut non nn lm gim vic to thnh glucose, gim hp thu glucose vo mu.Acarbose :Hnh1.6: Mt s i din ca nhmNhm 6 : Cc cht c ch DPP-IVCc thng tin v DPP-IV v cc cht c ch DPP-IV s c trnh by chi tit trong phn 2 v phn 3.1.3. Tc dng trong iu tr v so snh cc nhm thuc : 1.3.1. Tc dng trong iu tr ca cc thuc : Mi nhm thuc iu tr T2DM u c nhng cch tn cng v tc dng khc nhau. Nhng u c im chung l : Lm gim mc ng huyt .Gip ci thin kh nng s dng glucose ca c th .Lm gim cc triu chng tng ng huyt .Trong : Insulin : iu tr bnh T v cc bin chng ca n nh T typ 1, T typ 2 (khi cc bin php iu tr khc khng c hiu qu) v iu tr T thai k.Sulfonylure v meglitinid lm tng kh nng bi tit insulin t tuyn ty. Metformin lm gim lng sn phm glucose t gan v lm gim cc yu t khng insulin. Cc cht c ch -glucosidase lm cn tr kh nng hp thu glucose rut.Nhm thiazolidindion lm gim cc yu t khng insulin. Cc cht c ch DPP-IV lm tng kh nng gii phng insulin t tuyn ty ngay sau ba n.So snh u nhc im ca cc nhm thuc : u im Nhc imNhm sulfonylure (glyburid, glimepirid, glipizid) [23-25, 93]- Tn cng nhanh- Khng nh hng ti huyt p- Khng nh hng n LDL cholesterol- Liu tin dng- Gi thp- Nguy c trn tiu ha t hn metformin- Tng cn (trung bnh t 2.25kgs n 4.5kgs )- Tng nguy c h ng huyt- Glyburide c nguy c h ng huyt cao hn glimepirid v glipizidMetformin (nhm biguanid)- Nguy c h ng huyt thp- Khng lin quan ti vic tng cn- Tc dng tt trn LDL cholesterol- Tc dng tt trn triglycerides- Khng tc dng ln huyt p-Gi thp- C nguy c cao trn h tiu ha(bun nn v tiu chy)- Khng dng cho ngi c bnh thn va hoc nng v c nguy c tim mch- Km thun tin trong liu s dng hnCc cht c ch alpha-glucoside (acarbose,miglitol)- Nguy c h ng huyt thp hn sulfonylure- Khng nh hng ti vic tng cn- Gim lng triglycerid - Khng tc ng ln cholesterol- Km nht trong tt c cc thuc iu tr T dng vin trong vic lm gim HbA1c- Nguy c tc dng trn h tiu ha cao so vi cc thuc dng vin khc tr metformin- Liu dng thun tin- Gi caoNhm thiazolidindion (pioglitazon, rosiglitazon)- Nguy c h ng huyt thp- Cng c mt phn tc dng tt trn cholesterol (HDL)- Pioglitazin c lin quan n vic lm gim triglycerid- Liu dng thun tin- Tng nguy c tim mch- Tng cn (trung bnh cng t 2.25 n 4.5 kgs- C nguy c trong vic lin quan ti triu chng ph- C nguy c cao trong vic lin quan ti bnh thiu mu- Tng nhng tc dng khng tt trn cholesterol LDL- Rosiglitazon lin quan ti vic tng triglycerid v nguy c nhi mu c tim- Pioglitazon lm tng nguy c ung th bng quang- Kh nng tc dng chm-Gi caoNhm meglitinid (nateglinid, repaglinid)- Khng c tc dng xu no trn cholesterol- Tc tc dng nhanh- Repaglinid c lin quan ti vic tng nguy c gim glucose huyt v cng lm tng cn ging sulfonylure- Nateglinid km hiu qu hn trn HbA1c- Liu dng cha c thun tin- Gi caoCc cht c ch DPP-IV- Nguy c h ng huyt thp hn sulfonylure- Nguy c trn h tiu ha thp hn metformin- Liu dng thun tin (c th ung)- C kh nng duy tr v hi phc t bo [94]- Gim HbA1c km hn cc thuc tr i tho ng khc- Ch c gi tr tht s khi dng km vi cc thuc tr i tho ng khc- Gi caoPHN 2: C IM CA PEPTIDYL PEPTIDASE IV. MT MC TIU PHN T TRONG NGHIN CU PHT TRIN THUC IU TR TIU NG.2.1. Ngun gc v c im cu to : 2.1.1. Ngun gc:H DPP-IV l mt phn h ca prolyl oligopeptidase (POP), gm 4 enzym : DPP-IV, FAP, DPP-8, DPP-9 v 2 thnh vin khc khng phi l enzym : DPP-IV-like protein-6 (DPP-6, DPL-1hocDPP-X)vDPP-10(DPL-2).CcthnhvincahDPP-IVutinctXaa-Pro-v Xaa-Ala- ca cc dipeptide ( y : Xaa l bt k amino acid tr prolin) t N-tn ca cc protein [63].SkhcnhaucachnhccthnhvintronghDPP-IVlscmtca2aaglutamat nmtrongcchcxctc,mnhngchtnyrtcnthitchoshotngcaenzym[4]. Nguyn nhn DPL-1 v DPL-2 khng c kh nng xc tc c cho l do 1 s aa b th trong hc xc tc [37].Hnh2.7: c im v tr ca 6 thnh vin trong h DPP-IV,vi sDPP-IV l dng ha tan ca DPP-IV [10]Dipeptidayl peptidase IV (DPP-IV- E.C. 3.4.14.5) ln u tin c tm thy t gan chutnm1966[70,127].CngilCD26hayproteaseloiserinexuynmngtypIIhayenzym adenosine deaminase lin kt vi protein (ADAbp); l mt dipeptidase gm 766 acidamin, c tc rt xc tc nhanh bn trong t bo [40, 67, 75]. Mt dng ha tan ca enzym cng c tm thy trong huyt tng [47, 74].2.1.2. Cu trc ca DPP-IV : Hnh2.8: Chui theo hng ngang cc acid amin trn DPP-IV v POPGhi ch: [133] hDPP-IV v rDPP-IV : DPP-IV t ngi v chutHng trn l thng tin v cu trc bc 2 ca DPP-IVNhng mi tn mu vng v nhng thanh mu : ch biu din s khc nhau gia cc aaY : cc v tr glycosyl ha giu in tMu : cc v tr c kh nng glycosyl ha Nhng ng mu xanh vi cc cystein : cc lin kt disulphidMu hng : cc gc xc tcMu xanh da tri : biu din cho tyrosin c chc nng n nh nguyn t oxy m phnMu xanh l cy : cc aa nm trong lin kt ca adenosine deaminaseMu xm : phn xuyn mngCu trc tinh th (Hnh 2.9) ca vng ngoi (t aa 39 ti 766) ca DPPIV khong 2.1Ao vgm 2 phn chnh: 1 N-tn gm 8 -chn vt to thnh hnh dt (t aa 61 ti 495) v 1 C-tn c / hydrolase (t aa 39 ti 55 v 497 ti 766) [6, 104, 133].Hnh2.9: Ton th cu trc ca DPP-IV dng homodimerGhi ch : Dimer mu v xanh; v tr hot ng ca cc aa hnh cu v si, trc xon mu vng [6].Cchcvngchnvttvovngxctcthyphn,vbbacatrungtmxctc (S630; H740 v D708) nm ti v tr giao thoa gia 2 vng. Tc xc tc cho vic phn ct c cht H-Ala-Pro-7-amido-4-trifluromethylcoumarin (Ala-Pro-AFC) ca DPPIV l 5.2 * 106 M-1s-1

[6, 122].Mi n v bt i bao gm 2 homodimer [51]. Cu trc cc monomer (Hnh 2.10) ph hp vi lch tiu chun (RMSDs) l t 0.64Ao ti 0.98Ao cho cc nguyn t nng v t 0.28Ao ti 0.56Ao cho cc nguyn t C. B mt ca mi dimer bao gm: Si cui cng (8) ca vng trung tm c cu trc gp np. 2 xon cui cng (G v H). Quai nm gia 6 v E. i song (1* v 2*). 8 ch yu cha cc aa s nc hnh thnh nn tng tc vi b mt trung tm ca dimer. Xon -H hnh thnh lin kt hydro vi khp ni gia G v 8 trong 1 monomer khc. Cnh tay ca si i song c hnh thnh bi 1* v 2* cng tng tc vi cc cnh tay khc lin quan, G, v khp ni gia 6 v E trong monomer khc [11, 21, 41, 64]. Cu trc chnh ca DPP-IV gm 9 v tr glycosyl ha c lin kt vi N ti cc v tr: N85, N92, N150, N219, N229, N281, N321, N520, v N685 [5]. V tr m hnh chn vt c hnh thnh bi cc -chn vt, n gm c 8 khc nhau c lp li to thnh cu trc dt. Mi cu trc dt li gm c 4 si i song c cu trc gp np [51, 59].Hnh2.10: Cu trc cc monomer [51]2.1.3. Cch tn cng : Da vo cu trc v hot tnh sinh hc ca DPPIV, s c 2 cch tn cng vo cc v tr hotngcan(Hnh2.11):thngquavtrmcavngchnvthocvtrmbnnm gia b mt ca vng chn vt- v vng xc tc thy phn [51, 106] :Hnh2.11: Cch tn cng vo DPP-IVVtrmhnhchnvtlinktvi vnglncachuibnchnhthnhtbmt chung gia vng -chn vt v vng hydrase. L hnh cu ny to ra c hi th 2 tn cng vo v tr hot ng [51, 119].2.1.4. Cc v tr c bit trn DPP-IV : Chui peptid u N-tn c nhn bit bi cc aa tch in m trung tm hot ng ti E205vE206[26]vccnhbilinkthydrovichuibnca2aaglutamic.2aa glutamicnycginguyndngcutrcbilinktcumuivlinkthydroviaa R125, Y662, D663 v N710. Kh nng xc tc ca DPP-IV cho vic phn ct cc peptid tt nht l khi c 1 pro hoc ala ti v tr P1 [81, 83]. (cc tn gi c bit nh : P1, S1, S2..s c ch r trong gin c cht v c s thit k phn sau).HcsncS1gmccacidaminV656,Y631,Y662,W659,Y666,vV711.HcS2cnglsncvcxcnhbichuiacidaminR125,F357,Y547,P550,Y631,v Y666. GcSer630chnhthnhbiccaaGly-Trp-Ser630-Tyr-Gly.Chuinylcnthit cho kh nng hot ng ca DPPIV [108] v c lu li trong h / hydrolase (Gly-X-Ser-X-Gly). Hng ca S630 c gi bi nhng lin kt hydro nm gia nhm carbonyl ca S630 vi amide ca Y634, v amid ca S630 vi carbonyl ca V653 [6, 122].Trung tm hot ng ca DPP-IV :Trung tm hot ng ca DPP-IV gm b 3 xc tc: Ser630, Asp708 v His740. Ngoi ra, cn c cc v tr xc tc cn thit cho hotng ca DPP-IV nh:Tyr547 nm vng xc tc thy phn, Glu205 v Glu206 nm vng -chn vt [4, 99, 108, 122] .Trung tm hot ng ny nm mt khoang rng l b mt giao ca 2 vng : vng -chn vt v vng xc tc thy phn. Ser630 nmgia si 5 v trc xon C, hnh khuu v c tnh i nhn, mang c trng loi /hydrolase [109]. Nhm hydroxyl ca serin cng ha tr tt v to linkthydrovinhmimidazolcaHis740(2.6Ao).His740nm gia si8 v trc xon F. Vi mt khong cch 2.75Ao ti vng imidazol, mt trong nhng ion oxyanion ca Asp708 to lin kt hydro vi His740 v to thnh b ba xc tc. Ion oxyanion khc ca nhm carboxyl ca Asp708 c ta c xc nh bi 2 chui chnh l cc nhm NH (Val711 v Asn710) [109].Haiacidglutamic(Hnh2.12)thnghngvicccchtpeptid v vy ch nhng aa vi mtbnnhhn,nhlprolinvalanincthtmnvtrhotng.Cutrcnylm sng t ti sao DPP-IV u tin ct ti Xaa-Pro v Xaa-Ala v cc v tr c bit khc ca c cht [100].Hnh2.12: Cc v tr hot ng trn DPP-IVGhi ch : saxagliptin (mu ti) Acid gltutamic 205 v 206 hnh thnh cu mui vi nhm amin nm ti P2 ca cc cht c ch[4].CcnhmaminochatrongccchtcchDPP-IVnhalogliptin,saxagliptin, sitagliptinvvildagliptinuccholtngtcvi2glutamatnyvimnhcacc tng tc khc nhau. Cc tng tc ny u c chng minh khi kim tra cu trc tinh th bngX-quangcaphchpDPP-IVvisitagliptin[28],saxagliptin[100]valogliptin[53]. Trong saxagliptin c tng tc mnh hn so vi alogliptin, sitagliptin v vildagliptin v n c c tng tc vi Ser630 [100].Asparagin 710 v arginin 125 to lin kt hydro vi nhm carbonyl ca aa trn P2. Hc s ncnhclmytthnvinhngvngthmnh,cbitlvngpyrrolidinhoc phenylcachtcch.HaityrosinhnhthnhnnloxyanionvvthTyr547cxeml quan trng cho b ba xc tc [29].Ngoira,Gly628vGly632cngquantrngchosthnhlpcuhnhkhihngcho Ser630 vo ng v tr. iu ny cng ph hp vi nhng nghin cu trn chut c DPPIV khi chrachuiGly628-X-Ser630-Tyr631-Gly632 cngcnthitchokhnnghotngca DPP-IV [108]. tm hiu r hn v trung tm hot ng ca DPP-IV, chng ta s nghin cu dng phc ca DPP-IV vi c cht chn lc. Xt c cht chn lc l :diprotin A khi gn vo cc v tr hot ng ca DPP-IV (Hnh 2.13 v Hnh 2.14) :Vi diprotin A : Diprotin A (Ile-Pro-Ile), mt cht c ch cnh tranh [120, 121] , n c lin kt cng ha tr vi v tr hot ng Ser630 ca DPP-IV. u N-tn Ile (P2) v Pro gc (P1) c kh nng tng tc vi cc v tr lin kt ca c cht.DPP-IVlmthyphnccoligopeptidvproteintN-tn,ctccnvdipeptidkhi onth2lprolin,hydroxyprolin,dehydroprolin,pipecolicacidhoc alanin. Cc tiun v prolin ti v tr P1 ca diprotin A dng trans v khp vi cc v tr hot ng trn enzym. Hc S1 gm: Val711, Val656, Tyr662, Tyr666, Trp659, v Tyr631, k nc, c th c lm y bi prolin tt hn l alanin.iucnthitchoviclinktcacchtlN-tncaccht,ncthcbo v hoc khng c bo v. Phc hp diprotin A cho thy nhm NH3+- tn c gi v tr rt chnhxcbitngtcmnhvicarboxylatcacpphntGlu(Glu205vGlu206)cng nhOH-caTyr662.Glutamicth3,Glu204,cmtlinkthydroviNHcaArg125, His126 v Ser127 cng nh nhm OH- ca Ser127. Hnh2.13: Diprotin A (IIe-Pro-Ile) v cc v tr hot ng ca DPP-IVGhi ch : C cht diprotin A c gi trong mt hnh t din theo lin kt cng ha tr vi Ser630. Cc lin kt mu xanh da tri ti : protein.Xanh da tri sng: phi t Xanh l cy : v tr hot ng ca Ser630. Nhng ng nt t : cc lin kt hydro.Hnh2.14: ng vo v tr hot ng ca DPP-IVDPP-IV l mc tiu phn t trong iu tr T2DM DPP-IV cng l enzym mng ph bin, cng c vai tr iu chnh cc qu trnh sinh l nh dinhdng,traoicht,mindchvnitit..trongquantrnglnhhngtiscn bngglucosenimi[19,42,46,111].DPP-IVxctcthyphnccdipeptidtiN-tnt polypeptid vi prolin hoc alanin v tr p cht [100, 119].C lin kt chn lc vi cc c cht cha prolin v tr P1, do nhiu cht c ch DPP-IVcdvng5cnhmphngcutrcprolinvdnhpyrrolidin,cyanopyrrolidin, thiazolidinvcyanothiazolidin[8].Ccchtnythnghnhthnhlinktcnghatrvi Ser630 [90].DPP-IVckhnnglmbthotccincretin[65],cbit l GLP-1 v GIP, u l cc hormon t rut c bi tit p ng vi lng thc n [66, 69, 116, 119].Hnh2.15: V tr cc aa trn GLP-1 v GIP [69] Hnh2.16: V tr tc dng ca DPP-IVHnh2.17: C ch hot ng ca GLP-1 trong vic iu tit glucose.Ngay sau khi c gii phng t nim mc tiu ha trong cc ba n, cc hormon incretin (GLP-1vGIP)thamgianhchthtrgipinsulinctitratcc t bo o ty lm gim lng glucose [119, 137].Hnh2.18: Hai cch bo v dng hot ng ca hormon incretin trc tc dng ca enzym DPP-IV [65]Khi GLP-1 nhanh chng b thi tr (thi gian bn thi khong 1 pht) v chng b phn ct bi DPP-IV thnh dng mt hot tnh, c cc nghin cu c a ra nh s dng cc GLP-1 ngoi sinh, cc GLP-1 protein nhit hch v cc cht tng t GLP-1 khng DPP-IV iu tr T2DM.DngGLP-1chobnhnhnditruynmntnhdntivic gim ng k c lng glucose trong mu v huyt tngHbA1c [39, 148].Cc nghin cu lm sng trn ngi cng a ra cc bng chng chng minh hiu qu ca vicgimglucosehuytkhidngsitagliptinvvildagliptinnchockthpvicctc nhnchngTkhchinc.CchtcdngmccchtcchDPP-IVlmgimng huyt l khc bit so vi cc tc nhn lm h ng huyt khc. Chng kim sot lng ng trongmucaobivicgychotuyntytngtitinsulin,tngtunhonGLP-1,gimtit glucagon v truyn tn hiu cho gan gim sn xut glucose [103]. nh gi ny nhn mnh sphttrincaccchtkhcnhaucchDPP-IVdatrnccnghinculmsngvtin lm sng. N bao gm c tim nng pht trin cc cht c ch DPP-IV theo tc dng ca GLP-1 [66].Ngoi ra, cc th nghim trong lm sng ca vildagliptin i vi nhng ngi b T2DM ctinhnhkhidngnliuhaydngkthp vi metformin, pioglitazon hoc insulin v cngchngminhckhnngcithinlngnghuyt [101, 118, 123]. Nhng bnhnhnciutrviliu100mgvildagliptin/ngytrongkhong24tunchothymcgim A1C ( trong , A1c l t l phn trm ca HbA1c ( gn glucose) trn tng Hb ton phn, gip nh gi lng ng trung bnh c trong mu trong vng 2-3 thng gn nht bng cch o nng caglycosylatedhemoglobin)trungbnhl1.1%tmcbanul8.7%trn1.301bnh nhn [105]. Khi dng kt hp vildagliptin vi metformin cng ci thinng k kh nng kim sot ng huyt v tt hn so vi dng kt hp metformin v gi dc. Sau gn 1 nm dng kt hp vildagliptin vi metformin; 41,7% cc i tng t mc A1C l 7% (t mc A1C ban u l7.6-7.8%).Ngcli,ch10.7%bnhnhndngmetforminkthpvigidccA1C 100 M. Mt vi cht c ch u tin c cng b l c ch DPP-IV km chn lc trn DPP-8/9. Do , n cng cn ht sc thn trng khi tip tc pht trin nhm c ch DPP-IV trong nghin cu iu tr T2DM. Mc d c ch cha c hiu rnhngFDA(U.S.Foodv Drug Administration) yu cu tt c cc cht c ch DPP-IV phi c th trn kh [98].Hnh2.20: Tnh chn lc trn DPP-IV ca mt s cht c ch.PHN 3: CC CHT C CH DPP-IV. 3.1. Cht dn ng* Theo phng php sang lc t nhin, c 2 cht l:TMC-2(3.1),phnlptAspergillusorzaeA374(IC50=7,7M)[147].Mtcht ctnghphahcvikchthcnh hn (3.1) v xc nh c cu trc l TSL-225 (3.2) vn duy tr c hiu lc tng ng (IC50 = 5,7 M).Sulphostin (3.3) c phn lp t Streptomyces sp MK251-43F3 [12].* Theo phng php sng lc ngn hng ha cht (HTS), c cc cht l:Ccchtdnng(Hnh3.21)gm:cyanopyrrolidin,noncyano-pyrrolidinvthiazolin, xanthin v -phenethylamin [4, 20, 72, 77, 95, 117, 126, 130, 131, 138, 141].Hnh3.21: Cc cht dn ng trong tng hp ca DPP-IV.*Theo phng php thit k c ch ch, c (3.6), (3.7), (3.8) : Hnh3.22: Gin c cht ca DPP-IV.Nhngchtcchutinxuthinvocuinhngnm1980 v u nm 1990. N l c s quan trng cho vic nghin cu SAR (nghin cu lin quan gia cu trc v tc dng) cho nhng kho st sau . Cc cht c ch DPP-IV c chia lm 2 nhm chnh : nhm cng ha tr v nhm khng cng ha tr.Tripeptide diprotin A (Ile-Pro-Ile (3.4)) v diprotin B (Val-Pro-Leu (3.5)) l nhng c cht c ch DPP-IV tng ng vi IC50 l 8 v 920 M. Hai cht ny u cha proline v tr im nt.Lys[Z-4-NO2]-pyrrolidin(3.6)vimtvngthmkhccgnvoP1,nimDPP-IV thng tn cng vo prolin v alanin vi IC50=2 M. Hai hp cht khc vi vng thm khc gi proline ti v tr P1 l Val-pyrrolind (3.7) v Ile-thiazolidid ((3.8), P32/98) vi IC50 tng ng l 6.0 v 2.8 M. Nhng hp cht ny tr thnh c s cho vic pht trin cc cht c ch DPP-IV.Hnh3.23: Cc cht c ch nhm peptid v khng cng ha trCcchtcchDPP-IV c lin kt cng ha tr vi DPP-IV cng c nghin cu sm. N-peptidyl-O-aroylhydrolamin(3.9)lchtcchyu,cchkhngthunnghchvdb thy phn [44]. Cc phosphonat dipaptid cng l cc cht c ch khng thun nghch .Hnh3.24: Cc cht c ch cng ha tr.Cyanopyrrolidin : Trong cc cht dn ng trn th cc dn cht ca cyanopyrrolidin chim s lng nhiu nht v c nhiu thuc i vo s dng hin nay. Vic tm ra cht dn ng cyanopyrrolidin u tin (cyanopyrrolidid, pyrrolidin-2-nitril, prolinenitril) nm 1995 c da trn 2 c s: Th 1 :Trong vi thp k nay chng ta bit, cc nitril peptid c kh nng c ch nhm cystein proteasevchngcngckhnngcchyunhmsterineproteasevnckhnngto imidat vi gc xc tc serin [85, 89, 134, 143].Th 2 :Khongcuinm1980vunm1990ccdnchtcaaminoacylpriloncnhmi in t tng tc vi gc xc tc serin c cng b v c cho l c tc dng c ch DPP-IV vimcthigianvtcgitng ng l Demuth nm 1989 [43, 54], Flentke [38, 57] 1991v Powers nm 1994 [22, 33, 82, 128] (Hnh 3.25), y c xem l cc cht c ch DPP-IV u tin [34]. Tuy nhin, cc hp cht trn u c hiu lc iu tr thp v n nh ha hc khng cao.Hnh3.25: Cc cht dn ng ca cyanopyrrolidin.Quan st cc d kin trn, Sherwin Wilk v cng s tm cch gn nhm nitrile vo cc chtcchDPP-IV:aminoacylprolin.Ktqulhnhthnhccdnchtcacyanopyrrolidin chiulciutrtt,nnhhahccaovcngctnh chn lc trn enzym ny [18]. Tip nhng nm sau , Ferring c 2 bi bo khi nghin cu cc cht c ch DPP-IV da trn cuhnhcacyanopyrrolidin,htcbngsngchnm1993vcxutbnnm 1995 [13, 87] :Th 1 : gn thm cycloalkyl lm tng c hiu lc v bn ha hc [13, 115].Th2:araccnghincuSARvivtrP1.Vivngno5v6cnhcha kt hp nitrogen, sulfur v oxygen, mi nhm u c nitril ti v tr 2 [13, 115]. Hp cht c hiu lc tt nht l thiazolidid.Cc hp cht t Ferring cho thy kh nng c ch vt tri, v n nh ha hc cao. VicnghincuDPP-IVnhlchiutrcaT2DM lm cho nhm cyanopyrrolidin tr thnh ti nghin cu v cng rng ln trong sut nhng nm sau . Cht in hnh u tin l NVP-DPP728(nm 1998) [72, 96, 141]. Cht ny c chng minh c hiu lc tc dng trn ngi [9] v sau l vildagliptin [140] cng saxagliptin [15, 61] l nhng cht tin b nht hinnaytrongthhcyanopyrrolidin.Vildagliptin v saxagliptin u c pht trin thnh thuc trn th trng.Nhng c cht hoc cc dn xut tng t c cht ((3.18)-(3.21)) c ch DPP-IV u tin l [32, 43, 55, 68, 80, 84, 139] :Mttrongnhnghpcht(vd(3.20)v(3.21))lnhngchtcchkhngthun nghchcaenzymvcthitktorasnphmcnghatrviserinchottnhxc tc. Chnh v th to ra cc hp cht c hiu lc cao, bao gm cc cht c c hot tnh trn in vivo [22].Cchpchtctmthysmkhcgmccdnxutcaacidproline-boronic, nh l ((3.22)-(3.23)-(3.24)).Nhnghpchtnyuth hot tnh c ch trn DPP-IV, bi v ngi ta cho rng acid boric c lin quan n vic c ch enzym ny [57]. 2 trong s 3 cht trn l n nh v mt ha hc [76] v chn lc [38, 71, 129] .Cc acylthiazolidid v acylpyrrolidid l nhng c cht gi [17, 50] . Cht (3.25), P32/98 c kim tra hot tnh v n cng c kh nng ci thin lng ng huyt [16, 117]. Cu trctinhthcavalin-pyrolidin(3.26)chothyvngpyrrolidinnmtronghcsncS1, trongkhivanilechuibnlikhngcbtktngtcnovienzym[122].iuny khin cc nh khoa hc nghin cu ra (3.27) c hiu lc cao hn. Hp cht (3.27) chim khng gian hc S2 tt hn v hiu lc iu tr cao hn. Tuy nhin (3.27) l phn cc v pK khng cao v c ci tin thnh (3.28) c pK cao hn [110].Nhnitrn,viccchchnlctrnDPP-IVlrtquantrngkhiiutrtrn ngi v l do an ton. Mt nghin cu vi nhng cht chn lc cao bt u vi (3.29) v (3.30),theo cu hnh threo- (3.29) cho thy kh nng chn lc tt hn cu hnh erythro- (3.30) t 5 n 10 ln, (3.31) l 1 cht c ch chn lc trn DPP-IV (IC50 = 3nM ) v khng c ch knh hERG. Trong cc cht , cc hp cht c mt proton trong P2, nh l (3.31), c pK thp nhng khng c ch knh hERG, trong khi cc hp cht nh l (3.32) thiu proton c pK hp l nhng c ch hERG[146].Ccchttrongdytrnuclinktmnhviprotein[49].Pyridon(3.33)c tnhchnlcvcktqutt,nghincuSARlinquanticcpyridinctinhnh [145]. Vic ti u ha (3.34) c tc dng c ch chn lc trn DPP-IV vi IC50 = 34nM, DPP II (IC50=8M),DPP8vDPP9(IC50>50M).Nkhng c ch hERG v c pK hp l. Trn chutgy(khngm),(3.34)chothttcdngngungtt(OGTT)vi1mg/kg.Ty chng ta c th thy:Th1:Quanstthyskhcnhaugiaccdngngphn(3.29)v(3.30)cskhc bit trong vic c ch DPP-IV v DPP8/9 l iu quan trng nghin cu tip.Th2:Trongsccchtny,dythnhtc thng s pK hp l nhng khng an ton cho ngi c nguy c tim mch, trong khi dy cht th 2 th ngc li.Th 3 : n nh ca cc hp cht : vichng n c c thng s pK hp l v an ton vi ngi c nguy c tim mch hng ti cc hp cht mi. Cc cyanopyrrolidid (cc hp cht ny c ch cc enzym trong nhm prolyl oligopeptidase), nh l (3.35) v (3.36) c kh nng c ch DPP-IV [86] . Cc cht ny c lin kt thun nghch nhng hnh thnh lin kt cng ha tr vi Ser-630 ti v tr hot ng ca enzym, iu ny c ch r trong cc nghin cu cu trc bng X-quang [51, 107]. Novartis ti u ha nhm ny v c 2 sn phm c th hot tnh lm sng (3.37) v (3.38) [140, 141].Hp cht (3.39) (DPP-IV IC50=1.9nM) v (3.40) (IC50= 11 nM) c tm ra bi hng Merk. Ti u ha (3.39) a n (3.41) (IC50=119 nM), c sinh kh dng ng ung thp trn chut (3%), trong khi ti u ha (3.40) ti (3.42), cng c sinh kh dng ng ung thp hn [36, 144]. Cc nghin cu trn chuyn ha cho thy vng piperazin c th b oxy ha. Vic thay i vng piperazin to ra (3.43) (MK-0431, sitagliptin, IC50=18nM), c tc dng c chn lc tt trn DPP-IV, v sinh kh dng ng ung trn chut l 76%. Hp cht ny tr thnh thuc. Cu trc tinh th ca (3.43) cho thy carbonyl amid vi vng phenyl nm trong hc S1 [77].Hp cht (3.44) (IC=0.5 nM) l dn cht t (3.39). N khng c ch hERG nhng c sinh khdngkmvkmhpthu.(3.45)(IC=20nM)csinhkhdngttnhnglchtcch hERG (hERG IC50=870 nM ) [48].Cht (3.49) c tm ra bng phng php HTS IC50=10 nM. Ti u ha hiu lc c (3.47),viIC50=0.1M,vihiulctnggp100.000ln.Cutrctinhthca(3.47)cng c nghin cu r [114]. Hp cht lin quan ti nhm ny l (3.48) (IC=10 M) c kh nng ha tan cao, kh nng thm qua mng tt v chuyn ha n nh trong gan ngi, nhng n lm gimlngphospholipidvcchCYP3A4(IC50=5,4M).Sau,ccithinthnh (3.49)khngnhhngtiphospholipidvIC50choCYP3A4l30nM.(3.48)lhpcht tng t nhng thun tin hn v mt ha l [113].3.3. Phn loi cc cht c ch DPP-IVC hai cch phn loi cc cht c ch :Cch 1 : Da vo tnh gi peptid.Cch 2 : Da vo tnh cng ha tr.3.3.1. Phn loi theo cch 1 : Cc cht c ch DPP-IV c chia thnh 2 nhm : * Nhm gi peptid, trong nhm ny li chia thnh 2 nhm nh na, gm : + Nhm : M hnh cu trc gn ging glycin (Hnh 3.26) Hnh3.26: nhm Trong dy , da vo s c mt ca nhm th (Z) ti v tr C-2 ca vng pyrrolidine, dy c th chia thnh 2 nhm nh tip theo (Hnh 3.27) :CcchtcchkhngthunnghchkhiZ=diphenylphosphonateester[-P(O)(OPh2)] hoc O-acylhydroxamic acid (CONHOCOR)Ccchtcchthunnghchkhi(Z)=boricacid[B(OH)2],nitrile(CN)hoc hydrogen.Hnh3.27: nhm thun nghch v khng thun nghch+ Nhm : M hnh cu trc gn ging alanin. Cc dn xut ca piperazin MK-0431 (Hnh 3.28) Hnh3.28: Sitagliptin* Nhm khng gi peptid (Hnh 3.29) :Hnh3.29: Mt s i din nhm khng gi peptid.Phn loi theo cch 2 : DavoccnghincucutrckhnggiantX-quangcthchiaccchccchtc ch DPP-IV thnh 2 nhm[126] : Nhm khng cng ha tr : Gm sitagliptin, LY 2463665 v cc cht khc trong (Hnh 3.23).Nhm cng ha tr: Gm vildagliptin, saxagliptin v cc cht khc trong hnh (Hnh 3.24)3.3.3. Tm tt cc cht c ch DPP-IV trong th nghim lm sng v nghin cu :Bng 3.1 : Cc cht c ch DPP-IV trong nghin cu v th lm sng [7, 66]:Cc cht c ch DPP-IVTn thng gi/(Bit dc)Cng ty DPP-IV IC50Dng c ch v khong thi gianMK-0431Sitagliptin(Januvia TM )Merck & CoIC50=18nMKi=9nMc ch ngc v thi gian diLAF-237Vildagliptin (Galvus)NovatisIC50=3.5nMKi=17nMLin kt ng ha tr v c ch ngcBMS-477188Saxagliptin(Onglyza)Bristiol-MyersSquibb/AstraZeneca/Otsuka PharmaIC50=3.37nMKi=0.6nMLin kt ng ha tr v c ch ngcSYR-322(TAK-322)AlogliptinTakeda Pharmaceuticals,San DiegoIC50=7nMc ch ngcGSK-823093C Denagliptin Glaxo Smithkline Ki=22nM -NVP DPP728 Novartis AGIC50=22nMKi=11nMP32/P98Isoleucine thiazolidine DPP-IVProbiodrug AGIC50=420nMKi=80nMc ch ngcGRC 8200 MelogliptinGlenmark pharmaceuticals LtdIC50=1.61nM-PHX-1149Dutogliptin tartratePhenomix Corp - c ch ngcR-1579 CarmegliptinF Hoftmann-LaRoche Ltd & Chugai Pharma- c ch ngcGhi ch : - = cha xc nh 3.4. Cc thuc in hnh : alogliptin, saxagliptin, vildagliptin v sitagliptin. Hin nay, c 4 thuc c ph duyt lu hnh l: alogliptin, sitagliptin, vildagliptin v saxaglitin.3.4.1. Alogliptin (4.1) : Tn khoa hc : 2-[[6-[(3R)-3-amino-1-piperidinyl]-3,4-dihydro-3-methyl-2,4-dioxo-1(2H)-pyrimidinyl] methyl] benzonitril [52, 53]. CTPT : C18H21N5O2. KLPT : 339.39 3.4.2.1. Lch s nghin cu alogliptin : Cc dn cht ca nhm xanthin chnh l cht dn ng ca alogliptin. Ngi ta da vo cu trc nhm xanthin, nhm c nh gi l c kh nng c ch DPP-IV v cng nhn thy cu trc ca cc imidazol v uracil c cu trc gn ging nhm xanthin trn. V vy ngi ta thit k cc nhm chc nh aminopiperidin hoc nhm benzyl vo v tr ortho gn trn imidazol v uracil tng t nh xanthin cng nhm to c ch DPP-IV [27].C th, hng Takeda/Syrrx th nghim v tng hp hng lot cc dn cht ca nhm uracil hng c ch DPP-IV.Bt u t mt dn cht ca uracil l 6-chlorouracil. Da theo nghin cu ca Ferring : vic gn thm cc cu trc vo u nito s lm tng khng gian, ng thi khi c thm nhm cyano tng tc vi enzym tt hn v bn ha hc s cao qua s lm tng tc dng ca dc cht. Hng Takeda phi hp nhng nghin cu v th nghim gn thm nhm aminopiperidin.

Sau nhng th nghim v gn ti cc v tr khc nhau, ngi ta tng hp v tim ra cht c hiu lc cui cng tt nht l SYR-322 vi IC50 = 4 nM. Cht ny sau ny tr thnh thuc v mang tn alogliptin.Alogliptin cho thy kh nng c ch DPP-IV vt tri v tng tnh chn lc, hn 10.000 ln so vi cc protease lin quan nh DPP-8 hay DPP-9. N cng khng c ch cc enzym CYP 450 v khng chn kn hERG nng trn 30 M [53]. Hng dc phm Takeda th lm sng giai on 3 cho kt qu vo thng 1/2006. Sau khi c nhng kt qu tch cc t th lm sng giai on 3, Takeda gi h s ng k thuc mi ti FDA vo thng 12/2007. V thng 10/2008 n chnh thc tr thnh thuc [66]. Alogliptin cng c chp thun ti Nht thng 9/2008. FDA chp thun alogliptin nm 2013 vi 3 dng trong th trng l [79] : a) Tn thng mi: Nesinab) Dng kt hp vi metformin tn :Kazono c) Dng kt hp vi pioglitazon tn : Oseni.3.4.2.2. Chu trnh tng hp :Chng ta s nghin cu chu trnh tng hp mt dn cht ca alogliptin : Tm tt [78]: Victnghpcthchinthngquahaichttrunggianquantrngtrong,cmt dn xut ca aminopiperidin mang vng spirocyclic trnphn t piperidin. Chun b cht trung gianaminopiperidincthchinbngcchxydngvngcyclopropyltrckhignln vngpiperidin.Datheohngnghincunychngtacnchun b cht c ch DPP-IV c cu trc C mang 1 vng spirocyclic trn piperidinyl.Hnh3.30: C s thit k cu trc C da vo Alogliptin (A) v cc cht c ch DPP-IV (B)Chu trnh tng hp gm 2 giai on chnh : Giai on 1 : to ra (4.10) lm nguyn liu trung gian trong qu trnh tng hpGiaion2:tnguynliutrunggian(4.10)ngitatiptctnghpradnchtca alogliptin (C) nh s : Cc v tr tc dng ca C trn DPP-IV:Hnh3.31: Nhng v tr tn cng ca C vi DPP-IVNhm cyanobenzyl ca hp cht C c th lm y cc ti S1 (c hnh thnh t V656, Y631, Y662, W 659, Y666 v V711) v tc dng vi Arg125.NhmcarbobyltivtrC-2cthto mt lin kt hydro quan trng vi Tyr631 qua NH.Ngoi ra, vng cyclopropyl c th chim khong khng gian gn khi lm tng nhng tng tc k nc vi DPP-IV.3.4.2. Saxagliptin (5.1) : Tn khoa hc : (1S,3S,5S)-2-[(2S)-2-amino-2-(3-hydroxy-1-adamantyl)acetyl]-2-azabicyclo[3.1.0]hexane-3-carbonitril.Cng thc phn t : C18H25N3O2 .Khi lng phn t : 315.41 3.4.2.1. Lch s nghin cu saxagliptin :Cc nh nghin cu ti Bristol-Myers Squibb tm ra rng vic tng kch thc khng gian ca u N trong chui acid amin s lm tng sinh kh dng v n nh. Da vo gin c cht ca DPP-IV v phng php thit k c ch ch : phi kt hp ngn hng ha cht nhm chn lc ra nhng cht c hot tnh tc dng cao. Ngi ta thit k gn thm nhm cyclo alkyl vo vng prolinnitril nhm pht trin vng P2. Sau nhiu ln tng hp v th nghim vi cc cu hnh khc nhau : th cu hnh cis-4,5 c hot tnh tt [91]: Ngi ta tip tc nghin cu ti nhnh tc v tr R2 ca gin c cht nhm ti u ha tc dng [3]: V cui cng, ngi ta cng tin hnh th gn thm nhm adamantyl v c nhm OH ln adamantyl th c 1 cht c hiu lc tc dng cng nh n nh ha hc tng ln rt nhiu, k hiu l BMS-477118. V sau ny, sn phm mang tn saxagliptin (Onglyxa) [115]. Thng 6 nm 2008, AstraZeneca v Bristol-Myers Squibb trnh mt thuc mi vi tn bit dc Onglyza ti M v ng k lu hnh ti chu u [15]. N c chp thun bi FDA ti M vo thng 6/2009 cho Onglyza 5mg v Onglyza 2.5 mg. Sau , n c kt hp vi Once Daily Metformin XR v cng c FDA cp php vo thng 1/2011 di tn thng mi l Kombiglyze XR.3.4.2.2. Chu trnh tng hp : * Phn tch : Nhnthy,khivictng hp saxagliptin (5.1) quy m thng mi s t 2 dn xut ca aa l (5.2) v (5.3) theo s [125] :Saukhiloibnhmbovca(5.3)thctlica(5.1)chnhthnh bi cc aa l (5.2) v methanoprolinamid (5.4) theo s : Khi loi nc ca amin u tin v nhm bo v ca amin th c c saxagliptin (5.1).* Phn ng tng hp: gm 2 giai on Giai on 1 : Tng hp ra (5.7) t (5.2) theo s : Giai on 2 : Tng hp ra (5.1) t (5.7) theo s : 3.4.3. Sitagliptin (6.1) : Tn khoa hc : (R)-4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-amin.CTPT : C16H15F6N5O KLPT : 407.314 3.4.3.1. Lch s nghin cu v pht trin trn th trng : Da vo nhm cht dn ng l cyanopyrrolidin v thiazolidid phi kt hp vi gin c cht.

ng thi, vi nhng thnh cng t saxagliptin l tp trung pht trin nhnh -amino amid v trn prolin amid, ngi ta tip tc nghin cu hng ny v nhn thy nhc im ca cc ng phn saxagliptin l tn cng c vo melanocortin 4 receptor (MC4-R) . Da theo nhng nghin cu lin quan cu trc tc dng, ngi ta gn thm nhm phenethylamin v floro ha c vng phenethylamin. N lm tng hiu lc ln gp 2 ln [35] : V cui cng, ngi ta tip tc th nghim v gn thm nhm F ln nhnh benzyl th c sitagliptin (khi thay R = F ) vi hiu lc tc dng tt nht IC50 = 18 nM. Vo thng 10/2006, sitagliptin tr thnh cht c ch DPP-IV u tin c FDA chp thun trong iu tr T typ 2 [60]. Nghin cu cu trc tinh th ca sitagliptin tip tc c s dng sau . Mt cht c ch DPP-IV c hiu lc tt, tnh chn lc cao v sinh kh dng ng ung tt c tm ra bi vic thay th cyclohexylamin trung tm ca sitagliptin bng 3-aminopiperidin. Cht thay th 2-pyridyl trong nghin cu SAR cho thy vai tr quan trng trong hiu lc v tnh chn lc trn DPP-IV [127].NghincucclinktcasitagliptinviDPP-IVdidngcutrctinhthchothy [112].1. Nhm trifluorophenyl chim hc S1.2. Nhm trifluoromethyl tng tc vi chui bn cha Arg358 v Ser209.3. Nhm amino to lin kt cu mui vi Tyr662 v 2 nhm glutamate c carboxyl ha : Glu205 v Glu206.4. Nhm triazolopiperazin tng tc vi nhm phenyl ca Phe357.3.4.3.2. Chu trnh tng hp : tng hp sitagliptin, ngi ta i qua 2 giai on tng hp chnh : Giai on 1 : nhm to ra nguyn liu (6.8) t nguyn liu ban u l (6.2) theo s sau : Giai on 2 : t nguyn liu (6.8) c to ra trn ngi ta tng hp ra sitagliptin (6.1) dng mui theo s sau [77] : 3.4.4. Linagliptin(7.1) : Tn khoa hc : 8-[(3R)-3-aminopiperidin-1-yl]-7-(but-2-yn-1-yl)-3- methyl-1-[(4-methylquinazolin-2-yl)methyl]-3,7-dihydro-1H-purine-2,6-dion.CTPT: C25H28N8O2. KLPT : 472.543.4.4.1. Lch s nghin cu v pht trin trn th trng : Da vo nhm cht dn ng l nhm xanthin

Kt hp vi nghin cu c Ferring l gn thm vo u nito lm tng khng gian ca dc cht v tng tc vi enzym tt hn, ngi ta c hng lot th nghim v cng c hng lot cc cht c to ra [132]:

Cc nh nghin cu ti BI sau tm ra rng vic s dng nhm buty-2-nyl l mt nhm c hiu lc ln, c gi l BI-1356.Nm2008,BI-1356(linagliptin)quapha3lmsng,vccpphpthng5nm 2011 vi tn thng mi l: Tradjenta v Trajenta.Nghin cu cu trc tinh th da vo tia X cho thy rng cc lin kt nhm xanthin vi DPP-IV c nhng im khc bit so vi cc cht c ch khc [112] : 1. Nhm amino cng tng tc vi Glu205, Glu206 v Tyr662.2. Nhm buty-2-nyl chim hc S1.3. Nhm uracil to ra lin kt ri tng tc vi Tyr547.4. Nhm quinazolin to ra lin kt ri tng tc vi Trp629.3.4.4.2. Chu trnh tng hp : Btut(7.2) tc dng vi (7.3) to thnh hp cht (7.4).T (7.4) loi b nhm bo v to ra linagliptin (7.1).3.4.5.Ccthngtinlinquantidcnghc,khnngcchchnlcvhiulcin vivo: Bng 3.2 : So snh dc ng hc ca 4 thuc [66]Cc cht c ch DPP-IVCmax (M)CLp (l/kg/h)AUC(M.h/mg/kg)t1/2(h) F(%) loiTi liu trch dnVildagliptin 0.29 1.5 - 1.5 >90 CM [141]Sitagliptin 0.33*3.6 0.52 1.7 76.0 Chut [77, 135] Sitagliptin 2.2*0.36 8.3 4.9 100.0 Ch 135]Sitagliptin 0.33*1.68 1.0 3.7 68.0 CMAlogliptin 0.39@- - 2.7745.0Chut [52]Alogliptin 0.66@- - 3.04 68.0 ChAlogliptin0.46-9.6@- - 5.4672.0-88.0CMSaxagliptin - - - 2.175.0 Chut [15]Ghi ch : CL: thanh lc huyt tng, F(%): sinh kh dng ng ung, Cmax : nng ln nht trong huyt tng, CM: kh cynomolgus, * = ng ung c kim sot liu 2 mg/kg, # = ng ung c kim sot liu 5 mg/kg, @ = g/ml, - = ko c gi tr, ^ = ng ung c kim sot liu 10 mg/kg.Bng 3.3 : Hiu lc ca cc cht c ch DPP-IV vi cc enzyme khc c lin quan [66]Cc chtIC50 (nM) chn lcTi liu trch dnCc cht DPP-IVDPP8 DPP9 DPP-II FAPDPP8DPP9trch dnSitagliptin 18 4800>10000>100000>1000002667>5550[135]Vildagliptin3.5(Ki=3)Ki=810 Ki=95>500000NA 270 32[140]Alogliptin 7.0>100000>100000>100000>1000001428514285[52]Saxagliptin 3.37 244 104 >30000 NA 72 31[88]Ghi ch : NA : cha c gi trBng 3.4 : So snh hiu lc in vivo ca cc cht c ch DPP-IV: [66]Thi im ly mu (sau liu) (h)ED50(mg/kg)im ly mu (sau liu) (h)Saxagliptin Alogliptin Vildagliptin Sitagliptin7 0.1 0.1 0.5 2324 0.7 10 14 >30Ghi ch : Gi tr ED50 cho c ch hot ng ca DPP-IV trn chut cng Wistar ti nhng thi im khc nhau sau khi dngBng 3.5 : Thng tin thuc : Cc thng tin dng thucCc thucdng thuc Alogliptin Saxagliptin Sitagliptin LinagliptinTn bit dcKazano, Nesina , OseniOnglyzaJanumet, Januvia ( India) , Januvia TabTradjentaCh nh Chng tng ng huyt, dng iu tr T2DChng ch nhThn yu, nhim acid, mn cmKo dng cho nhng bnh nhn typ 1, lng keton trong mu caoMn cm vi thucng dng Thuc dng vin nn, dng ng ungLiu dngDng hng ngydng Liu khi pht da trn tnh trng bn nhnLiu ln nht c th l 25/2000 mgNgy ung 2 ln sau n, liu 1.2.5/500 mgLiu khuyn co l 2.5 mg hoc 5 mg/ln100 mg/ln 5 mg/lnThn trngSuy yu gan, vim ty,Trnh dng qu liuC tin s vi bnh HIV/AIDS, bnh thn,Trnh nhng ngi ang c nng alcohol cao khi ang dng thucThn yu, tin s vi TBnh nhn ang s dng insulin hoc ang dng thuc khc km theoLng hemoglobin A1C mc thpKhi dng thuc lng ng huyt ang mc thpD ng vi thuc, ngi gi, tr nh hoc ngi mang thaiTc dng ph Triu chng chung:Nhim trng din rng, bnh tim mch, sng vim, tiu chy, p huyt cao, au uNi mn, tng mc creatinin trong muau u, tiu chy, nn maTriu chng khc :Bun nn, nn, y hi, kh tiuau u, cc triu chng vim xoang, nhim trng nc tiuau c, khpBo qun Nhit phng 200C250CKn gi, xa tm tay tr em KT LUNSau mt thi gian tra cu, c, phn tch v tng hp cc ti liu tham kho, chng ti thu c cc kt qu sau: trnh by c c im ca dipeptidyl peptidase - 4 v ng dng trong nghin cu pht trin thuc iu tr tiu ng.DPP-IV l mt dipeptidase gm 766 acidamin, c tc rt nhanh bn trong t bo. Cu trc tinh th gm 2 vng chnh: vng -chn vt v vng / hydrolase. Da vo y gip ta tm ra 2 cch tn cng vo DPP-IV. Vic tm ra cc v tr c bit nh : hc s nc S1, hc S2 hay gc Ser630 v b 3 xc tc: Ser630, Asp708 v His740 l trung tm hotng ca enzym gip cc nh khoa hc tm ra cc nhm chc v nh hng tn cng su vo DPP-IV. 2. Trnh by c c s ha hc ca mt s cht c ch peptidyl peptidase - 4 hng dng lm thuc iu tr tiu ng c nghin cu pht trin gn y. Vic tm ra, c 2 cch phn loi c bn v quan trng cc cht c ch DPP-IV:gi peptid vkhnggiipeptidcngcnghatrvkhngcnghatr.ngthidavo2chtdn ngtrongtnhinvnhmccchtdnngtrongtnghp,cbitlnhm cyanopyrrolidin t nn tng cho c s thit k cc cht ng ch DPP-IV v t y m ra mthngnghincumigipphttrinhnglotccchtcchDPP-IVsaunynh sitagliptin, alogliptin, saxagliptin hay linagliptin c hiu lc cao trong iu tr lm sng. 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