• Nghien ciru - Ky thuat
^Nghien cihi tong hop toan phan erlotinib lam nguyen lieu cho thuoc
dieu tri ung tliu
Tran Ngpc Quyin'*, Tr4n Thi Cim Tu^ Bui Thj Thuy Hanh^
Nguyen Thi Hu•6^ag^ Nguyen Cmi Khoa', Phan Minh Itian' Nguyen Thi Phmmg', Ha Dang Cap^
' Vi^n Khoa hgc Vat lieu ifng dung, VAST
^Trucmg Dai hoc Ton Dik: Thdng, Tp Ho Chi Minh
^Tnrcmg Bai hpc Lgc Hong, Bien Hoa, linh Dong Nai 'E-mail: tranngocquyen979(^yahoo.com
Summary
Erlotinib, as a drug used in targeted drug delivery application due to its ability of travelling through cell membrane to inhibit cancer cell proliferation, leading to pre-established, programmed apoptosis of cancer cells and decreasing the chance of metastasis, was synthesized in form of Eriotinib hydrochloride (EH). By reference to many previous related reports, a synthetic procedure was established to get the intermediates and final product (EH) in higher yield. The obtained products, both the intermediates and final one, were structurally analyzed by NMR, A^S and quantitatively determined by HPLC. The purity of the obtained EH was quite high (98.2 %) and the total yield of the synthesis was desirable (42%). The synthesis procedure is subjected to further improvement for better yield and to enlarge the scale.
Keyvi/ords: Eriotinib, principle, anti-cancer drugs.
D^t vin de
Trong nhieu logi ung t h y n h y dgi trye trang, t u y i n tyy, t u y i n t i l n lidt, co t d cung, vu, phoi, thdn vd d i u CO, cde logi thy t h l tang trydng t l bdo bieu bi (HERs) ddng vai trd quan trpng trong t i l n t r i l n k h l i u ' ^ i . Thu t h l tyrosin kinase thuge gia dinh HERs la eae protein trdn mang bao t l bdo ed bieu hipn eao trong nhieu loai khdi u, bao g i m ca ung t h y phoi vdi te bdo khdng nhd (NSCLC). Trong dd qua trinh phosphoryl hod, cde phan tyrosin la giai dogn quan trgng trong d i n t r u y i n tfn hidu gdp phan eho te bdo phat t r i l n . Do dd, lam gian dogn s y phat then ede qua trinh d i n truyin tin hidu ndy se Idm dich d i n day t i l m nang cho d i l u tri c h l n g ung t h y . Cdc c h i t de e h l enzym protein tyrosin kinase hay ehat dc c h l qud trinh phosphoryl hod Id thuoc quan trgng eho d i l u tn ung t h y ed ehgn Ipe '^'.
Cde hgp c h i t 4-anilinoquinazolin d y g e b i l t Id nhdm e h i t cd hogt tinh d e e h l tot ede
enzymtyrosin thdng qua lien k i t ehpn Ipc vd thugn nghjch vdi vung tyrosin kinase k i t hgp HERs qua v\ trf adenosin triphosphat, ddn d i n dc c h l qua trinh phosphoryl hod eua tyrosin kinase trong t l bao ung thy. Mdt so ehat de e h l tyrosin kinase dyge phd c h u i n bdi Cyc Quan ly D y g c vd Thye pham My cho d i l u tn ung t h y phoi nhy erlotinib (thuoc Tareeva®) vd gefitinib (thulc Iressa®). Trong so dd, eriotinib the hipn hieu qua cao chong lai NSCLC d giai dogn di can sau khi t h i t bgi vdi it n h i t mdt phac do hod tri. Bdn eanh dd, Tareeva d y g e chdng minh ed it tac dyng phu so vdi ede logi thuoc trong phdc do hod trj truyen thong. Vdi nhdng y u d i l m tren, k l t d nam 1998, sau khi Cdng ty Pfizer INC cdng bo d y d i bao hd ban quyen ve quy trinh t i n g hgp vd hogt tinh tieu dipt te bdo NSCLC, r i t nhieu nhdm nghidn c d u da edng bo eae quy trinh eai tien de tong hgp hgp chat trdn hydng d i n san xuat mgt khi Pfizer INC h i t d y g e bdo hd ban quyen '^"^'.
TAPCHi DUgfC HQC - 7/2015 (S6 471 NAM 55)
Nghien cieu - Ky thuat
Cl Viet Nam, ehan doan phat hidn mdi nam khoang 150.000 ea ung t h y mdi, trong dd ung t h y phoi Id mdt trong nhdng loai pho b i l n nhdt '^1. Chi phi nhdp khau thuoc d i l u tri eho eac benh nhdn ung t h y la mgt ganh nang kinh t l cho eae quoc gia dang phat t r i l n nhy Viet Nam. Cae nghien edu h y d n g ddn san xudt ede loai thudc chong ung t h y la rat cd y nghTa d l giam chi phi nhap k h i u t h u l c vd eai thipn chdt lygng dieu tri eho bpnh nhan ung thy. Do dd, trong nghien
edu nay, chting tdi gidi thipu quy trinh tong hgp todn p h l n erlotinib hydrochlorid t d ethyl 3,4-dihydroxybenzoat (Quy trinh 1), tren eo sd tham khao ehgn Igc cdc phan dng tong hgp ed hieu qua cao d y g c cdng b l t r y d c dd '^^'. K i t qua dgt d y g e eua ehiing tdi cho thay eriotinib dygc dyng ed the tong hgp thdnh eong vd h y d n g d i n tong hgp trdn quy md idn lam n g u i n nguydn Ilpu san x u l t thuoc dieu tn ung t h y d Vipt Nam.
1 2 Cl o
,XX,
1 HCI Erlolmib hydrochlond
Quy trinh 1: Cac bwac trong quy trinh tdng hgp toan phan erlotinib
V a t l i e u , p h i f c n g p h a p v a t h u ' c nghiem
Hod chat va thiet bj Cdc hda chit
Ethyl 3,4-dihydroxybenzoat, tetrabutyl ammonium iodid, potassium carbonat, 1-ehloro- 2-methoxyethan, aeid sulfuric, aeid nitric, Pd/C, ammonium format, ethynyl anilin, 2-propanol, oxalyl chlorid va N,N-dimethyl formamid d y g c mua t d Cdng ty Acros Organies (BI) vd l\/1erck (Ddc). Aceton, hexan, formic aeid, tetrahydrofuran, ehioroform va ethyl acetat d y g c mua t d Cong ty Merck (Ddc). Cac tae ehdt khdng d y g c tinh c h l thdm.
Cac thiet bj phan tich cau true va do tinh khiet
Mdy cgng hydng t d hat nhdn 500 MHz Brucker Avanee 500; may sde ky Idng cao dp HPLC Agilent 1260 infinity LC system d i u dd DAD, egt
Eelipse C18 (250'<4,6 mm), kieh thydc hat pha tTnh 5 pm (made in USA) (Agilent Technologies/
USA); khoi pho d y g e do trdn mdy k h l i pho Agilent 6120 (Agilent Technologies/USA).
B l e u che cac chat t r u n g gian vd erlotinib Dieu che ethyl 3,4-bis(2-methoxyethoxy) benzoat (2)
3,4-Dihydroxybenzoat(2,0g, 10,8 mmol) dygc hda tan trong aceton (15 mL), potassium carbonat (4,6 g, 33,6 mmol) va tetrabutylammonium iodid (0.14 g, 0,36 mmol) tiep tye d y g e eho vdo hon hgp phan dng va khuay 20 phut d nhi^t dd phdng. Sau dd, them 1-chloro-2-methoxyethan (6,0 g, 5,8 ml, 66 mmol) vdo h l n hgp phdn dng, khuay dun hoan lyu trong 3 ngdy. Sau khi ket thuc, phan dng d y a v l nhidt dd phdng vd them mdt l y g n g diethyl ether. Hdn hgp phdn dng dygc Ipc de logi bd p h l n ran khdng tan, p h l n dung dieh qua Ige d y g e lam khan vdi Na^SO^, logi mau vdi than hoat tinh vd logi bd het dung mdi,
44
TAPCHi DlfQC HQC - 7/2015 (SO 471 NAM 55)
Nghien ciru - Ky thuat
thu d y g c c h i t dgng dau sdnh, mau vang nhgt.
Hda tan l y g n g d i u trdn trong ether ddu hda va lam lgnh d 0-5 °C de xuat hien chdt k i t tmh, dung mdi d y g e lam bay hai vd thu d y g e ehat rdn k i t tinh, mau trdng, hieu s u i t k i t tinh 80%, dp tinh k h i l l 95,4%. 'H-NMR (5, C D C y : 1,36 (t, 3H, CH3-CH3-); 3,45 (s, 6H, 2 x -OCH3); 3,78 (m.
4H. 2 x -CH^O-); 4,19 (m, 4H, 2 x CH^-O-); 4,32 (q, 2H, -CH2-CH3); 6,90 (m, I H , HAr, J = 8,5 Hz);
7,58 (s, I H , HAr); 7,65 (m, I H , HAr, J = 8,5 Hz);
"C-NMR (5, CDCI3): 14,38, 50,20; 60,78; 68,78;
70,94; 112,76; 115,14; 123,48; 123,98; 139,32;
152,94; 166,33. Anh h y d n g cua nhidt dd len hipu s u i t thu san pham d y g c khao sat.
Dieu che ethyl 4,5-bis(2-methoxyethoxy)- 2-nitrobenzoat (3)
Ethyl 3,4-bis(2-methoxyethoxy) benzoat (2,0 g; 6,72 mmol) d y g e hda tan trong AcOH (6,8 ml), HjSG^ (98%; 0,5 mL) va HNO3 (65%; 1,6 mL) d y g e them vao tgi 5 "C va khuay d eung nhidt do khodng 30 phut va 24 gid tai nhiet dp phdng. T i l n trinh phan dng d y a c theo ddi bang sde ky Idp mdng. Sau khi phan dng k i t thuc, hon hgp phan dng d y g c logi dung moi va thu d y a c ehat Idng dang d i u , san p h i m thd d y g c tinh ehe bang sde ki edt silica gel vdi hd dung mdi ethyl acetat: hexan (1:3 v/v) thu d y g c chat Idng, mau vdng, hipu s u i t dgt 87,28%; dg tmh k h i l t 95,6%.
'H-NMR (5, CDCI3): 1,36-1,38 (t, 3H, CHjCH^,
^J = 7,20 Hz); 3,38 (s, 6H, 2 x OCH3); 3,75-3,85 (m, 4H, 2 X CHp); 4 , 2 0 ^ , 2 7 (m, 4H, 2x C H p ) ; 4,31-4,42 (q, 2H, CH^, ^J = 7,20 Hz); 7,13 (s, I H , H J ; 7,50 (s, I H , H J . "C-NMR (5, C D C g . 20,12:
51,55; 59,92; 71,13; 76,18; 115,54; 118,03:
120,16; 142,24, 149,86; 150,12; 167,23. HRMS.
[M+Hr = 344,3, [ M + 2 H r = 345,1; [ M + H p ] * = 3 6 1 , 2 ; M L ^ = 3 4 3 , 3 ,
Dieu che ethyl 2-amino- 4,5-bis(2- methoxyethoxy)benzoat (4)
Dung djeh ammonium format (5,51 g, 87,5 mmol) trong n y d e (3,5 mL) d y g c cho vao hdn hgp 2-propanol (35 mL) va Pd/C (0,93 g) va khuay trong mgt phut de hoat hda Pd/C. Sau do, ethyl 4,5-bis(2-methoxyethoxy)-2-nitrobenzoat (3,00 g, 8,7 mmol) dyge cho vao hdn hgp phan dng vao khuay trong 20 phOt d nhidt do phdng. H l n hgp sau phan dng dyge Ipc va rda vdi h l n hgp ethyl acetat va 2-propanol, sau dd lam khan vdi
Na^SO^, logi bd dung mdi thu dygc chat Idng (2,51 g), hipu sudt 91,7%, ddtinh khiet 94,7%. 'H-NMR (5, CDCI3): 1,20 - 1.40 (t, 3H, CHjCHj); 3,43 (s, 6H, 2x OCH3); 3.4 (m, 4H, 2x CH^O); 3,53 - 3,54 (m, 4H, 2x C H p ) ; 4,40 (q, 2H, CH^Hz); 6,4 (s, I H , HAr,); 7,5 (s, I H , HAr). '^C-NMR (5, CDCI3):
16,14; 51,35; 58,93; 72,21; 74,39; 100,16;
102,31; 115,57; 139,28; 140,20; 148,27; 164,92.
HR-MS (methanol): [M+H]* = 314,2; [M+2H]* = 315,1; [ M + N a r = 336,2; MLT= 313.3.
Dieu che 6,7- bis (2-methoxy-ethoxy) quinazoion (5)
Ammonium fonnat (0,52 g, 8,1 mmol), ethyl 2-amino-4,5-bis(2-methoxyethoxy) benzoat (2,52 g; 8,1 mmol) d y g c hda tan trong fomnamid (3,80 mL) vd khuay, gia nhipt trong 2 gid trong khi quyen nitrogen d 160°C. Sau khi k i t thiic, phan dng d y g c dya ve nhiet dp phdng vd Igc bd p h l n c h i t ran khdng tan. Phan dung djeh cdn Igi d y g c ehilt vdi chloroform va nyde muoi, Idm khan vdi Na^SO^ va logi Isd dung mdi. San pham tho cdn lgi dyge k i t tinh igi trong diethyl ether vd thu d y g c k i t tiia la c h i t rdn quinazoion mdu vdng (2,01 g, hidu s u i t 87 %, dd tinh khilt 94,5%). Nhi$t do nong chay 190-19rC, 'HNMR{5, CDCI3): 3,48 (s, 6H, OCH3);
4,20^,30 (m, 4H, 2x GCH,); 3,85 (m, 4H, 2x OCH,);
7,60 (s, I H , HAr); 7,1-7,25 (s, 1H, HAr); 8,07 (s, I H , HAr). "C-NMR (5. DMSO-dg): 59.39; 71.32; 72,24;
112,81; 116,76; 140,23; 141,85; 145,11; 147,93;
152,64; 163,74. Hff-MS(methanol)[M+Hr= 295,1;
[M+2H]* = 296,3; [M+Na]* = 317,1; M^^ = 294,2. Anh hydng ciia nhiet dd len hieu s u i t phan dng d y g e khao sat.
Dieu che 4-chloro-6,7-bls-(2-methoxy- ethoxy)-quinazoiin (6)
6,7- bis(2-methoxy-ethoxy) quinazoion (1,87 g; 6 mmol) d y g c hda tan trong DMF (0,25 mL;
3 mmol) va CHCI3 (30 mL). Hon hgp tren d y g e nhd tdng gigt oxalytchlorid (1,60 mL; 2,42 g;
18,8 mmol) va dun hoan l y u trong 2 gid. Sau khi phan dng k i t thue, hdn hgp phan dng d y g e d y a ve nhidt dp phdng, loai bd dung mdi, c h i l t vdl n y d e va dichloromethan. Ldp hdu ea d y g c rda NaHCOj, lam khan vdi Na^SO^ va logi bd dung mdi thu d y g c ehat rdn (1,68 g), mau nau, hieu s u i t 82,6%, do tmh khilt 94,0 %. 'H-NMR (S, CDCI3); 3,50 (s, 6H, 2x OCH3); 3,80-3,90 (m, 4H. 2x OCH,); 4,32-4,35 (m, 4H, 2x OCH^); 7,26 TAP CHI DUQC HQC - 7/2015 (SO 471 NAM 55)
Nghien CIFU - Ky thuat
(s, I H , HAr),7,44 (s, I H , HAr); 8,89 (s, I H . HAr).
"C-NMR (5, CDCI3): 48,31; 68,33; 70,27; 103,01;
108,18; 120,12; 147,08; 149,44; 142,16; 156,37;
158,95. HR-MS (methanol): [M(35CI)+H]"= 313,1;
[M(="CI)+H]' = 315,1; [M(^CI)+2H]* = 314,3;
M(3'CI)+2H]* = 316,1; [M(^CI)+Na]* = 335,1;
[M(3^CI)+Na] = 337,1. Nhidt do nong chay : 107,5-108,5 "C.
Dieu che eriotinib hydrochlorid
4-Chloro-6,7-bis- (2-methoxyethoxy) quinazolin (1,00 g, 3,2 mmol) d y g e hda tan ^ o n g 15,0 ml nyde. Sau dd. 3-ethynylanilin (0,45 g.
3,84 mmoi), aeid hydrochloric (37%, 0,20 mL) d y g e eho vao hdn hgp phan dng va khuly, dun hodn l y u 40 "C. Phan dng d y g e theo ddi bang sac kif Idp mdng vd nhan thay k i t thue hoan todn sau 1,5 gid. Hdn hgp phan dng d y g e d y a v l nhidt dd phdng, Igc bd phan rdn, rda vdi n y d e vd h l n hgp ethyl acetat: n-hexan (1:1 v/v), ehat ran thu d y g c t i l p tyc d y a c say d 40 °C thu d y g e e h i t ran k i t tinh mau trdng (1,12 g), hipu suat 89,6%. San p h i m thd d y g c k i t tlnh Igi 2 lan trong MeOH, thu d y g e eriotinib vdi dp tinh k h i l t 98,2%. Sau khi thyc hidn cde bidn phap phan tfeh cau triie thi xac nhan san p h i m thu d y g c id eriotinib hydrochlorid. 'H-NMR (5, DMSO-d^):
11,13 (s, I H , NH); 8,70 (s, I H , HAr); 8,14 (s,1H, H-Ar); 7.70 - 7,81 (m, 2H, H-Ar); 7,28 - 7,51 (m, 3H, HAr); 4.31 - 4,32 (m, 4H, 2 C H p ) , 3,74 - 3,77 (m, 4H, 2 C H p ) ; 3,67 (s, I H , ethynyl); 3,32 - 3,34 (d, 6H, 2x OCH3). "C-NMR (5, CDCI3):
52,03; 69.79; 72,87; 79.86; 82,94; 98,79; 108,82;
112,06; 116,62; 121,45; 122,89; 123,22; 129,42;
142,86; 146,66; 151,38; 154,48; 158,42; 169,44 HRMS: [M{^C\)y= 428,0; [M(^^CI)+Hj* = 429,0:
[M(3'CI)+H]* = 430,9; M^^^= 429,9. Nhiet do nong chay: 226,5-228,0 -C.
K e t q u a v a b a n l u a n
San pham ethyl 3,4^is(2-methoxyethoxy) benzoat (2)
Trong s y hipn dien eua xuc tae ehuyen pha tetrabutylammonium iodid, hai nhdm hydroxyl cua ethyl 3,4-dihydroxybenzoat se ehuyen v l dgng phenolat vd tham gia phan dng S^2 vdi 1-chloro- 2-methoxyethan. K i t qua phan tfeh pho 'H-NMR eho thay ede tin hidu thu d y g e cxi dd djch chuyen hod hge tyang dng vdi eae proton trong eau tnJe
cua ethyl 3,4-bis (2-methoxyethoxy) benzoat. K i t qua nay eung phu hgp vdi n h i l u nghidn cdu trydc p*-^. D l i vdi pho '^C-NMR, cac tin hipu carbon ciing t y a n g dng vdi so carbon cua sdn phdm. Trdn co s d dd chiing tdi khang djnh rang san pham la ethyl 3,4-bis(2-methoxyethoxy) benzoat. K i t qua khao sat cho thay nhidt dO lam thay doi hipu s u i t cua phan dng, iSeu ndy cd t h l la do khi t i l n hdnh d 56 °C (hinh 1) cae tae chat de hda tan han, sy ehuyen ddng cua cde ion t y do nhanh han ndn ede tae c h i t de t i l p xuc, va chgm hi^u qud han d i n d i n hieu s u i t phan dng tot han.
Nbift dg pban mig (<!) Hinh 1 : Anh hwang cua nhidt dg Idn hidu suit
san pham thu dwtyc
K i t qua nghidn c d u cho t h i y dCi phdn dng d y g e thye hlpn trong khi q u y l n nitrogen vd thdi gian phan yng kdo ddi d nhipt dp sdi eua dung mdi, tuy nhien hipu s u i t ehf dgt 80% d dp tinh k h i l t 95,6%. O i l u ndy ed the do hinh thdnh sdn pham alkyl hda mdt nhdm phenolat vd do nhdm hydroxy benzoat bj oxi hoa c h u y i n hod thdnh quinon lam hon hgp sau phan dng cd mdu ndu ndn lam giam hipu s u i t eua phdn dng.
San pham 4,5-bis(2-methoxyethoxy)-2- nitrobenzoat (3)
Phan dng nitro hda hgp chat benzoat vdi tdc nhan than didn t d id ion nitronium (O^N*), ion ndy d y g e tgo ra bdi qud trinh proton hda ion nitrat (NO3) trong mdi t r y d n g aeid. Dung mdi phan dng la aeid acetic vd acid sulfuric nhy trong d i l u kidn phan dng da hinh thanh sdn p h i m vdi hipu s u i t va dg tinh k h i l t cao. Pho 'H-NMR eho thay cac tin hidu thu d y g c cd dg djeh c h u y i n hod hpe t y a n g dng vdi eae proton trong c l u tnJc cua ethyl 3,4-bis (2-methoxyethoxy)-2-nitrobenzoat, tuy nhien tin hipu proton d vj tri C ^ khdng cdn
TAP CHI DlTQC HQC - 7/2015 (S6 471 NAM 55)
Nghien CCFU - Ky thuat
xudt hien nhy trong pho d i a tac c h i t ethyl 4,5-bis(2- methoxyethoxy) benzoat. K i t qua pho nay cung phil hgp vdi nhidu nghien edu tryde ^^-^ . Pho MS eiJng chdng minh khoi lygng phdn t y eua san pham tao thanh la ethyl 4,&-bis(2-methoxyethoxy)-2- nitrobenzoat vdi cac pie the hidn khli lygng phan t d ddc tnjng t d san pham nitro hoa: [M+H]* = 344,3;
[M+2H]* = 345,1 va [ M + H p ] * = 361,2.
Sdn pham ethyl- 2-amino-4,5-bis (2-methoxyethoxy) benzoat (4)
Khd in situ mot each hidu qua nhdm nitro thanh amin bang ammonium format vdi xiic tac Pd/C Id p h y a n g phap mdi d y g c phat trien trong nhdng nam gan day P,4,5,9)_ y ^ j gy- hien didn eua Pd/C, NH^HCOO- se bj phan giai thanh CO^, H p va H nguydn t d de tham gia phan dng khd d y g c hap phy tren ehat mang xue tae Pd/C. Day la phyang phdp k h d thdn thipn. khdng sinh san pham phy, hidu suat ehuyen hoa cao va khdng c i n dimg cac t h i l t bj ehju d y n g d y g e dp sudt khf q u y l n nitrogen. San pham ethyl 2-amino- 4,5-bis(2-methoxyethoxy)benzoat tao thanh t d phan dng khd d y g e danh gid eau true bang pho 'H-NMR va pho MS Pho 'H-NMR cho eae tfn hipu the hipn s y djch e h u y i n hoa hge ciia mot so proton trong hgp c h i t amin tao thdnh t y a n g doi g i l n g vdi pho eua tde chat nitro. Tuy nhien, proton tham tren earbon lien ke earbon mang nhdm nitro thay doi dang ke do c h u y i n dich hoa hpe, t d 7.13 ppm (hgp chat nitro) den 6,41 (hgp chat amin). Dieu nay hoan toan hgp ly de nhdn bilt khae bipt eua vdng thom mang nitro vd mang amin la do nhdm nitro hiit dien t d mgnh, ddn ddn gidm hidu dng ehan eua cac didn t d xung proton nay, lam eho proton edng h y d n g d t d trydng thap. Khoi p h i MS cung the hidn cae pie the hien khoi lygng ion phan manh phii hgp vdi c l u triic eiia ethyl 2-amino-4.5-bis (2-methoxyethoxy) benzoat: [M+H]^ = 314,2; [M+2H]' = 315,1;
[M+Na]'= 336,2; va M^.^= 313,3.
San pham 6,7-bis(2-methGxyethoxy) quinazoion (5)
Quinazoion va dan xuat la cac hgp ehat dj vdng chda nita da d y g c chdng minh cd nhidu hogt tinh trong y d y g e . Cae hgp chat nay d y g c xem n h y la khung c a ban hoge t i l n c h i t eua
nhieu logi thudc chdng ung thy, khang viem, khang khuan, chdng dj dng, khang virus, ehdng sot ret, ehdng iao, tri tieu d y d n g , beo phi ''"'...
Do do hydng nghidn cdu tong hop quinazoion va ddn x u l t d l bao ehe cac ehdt dung lam thude d y g e dae bidt quan tam. Tong hgp quinazoion t d eae hgp ehdt aminobenzoat, ammonium format va formamid la mdt trong nhyng phyang phap pho b i l n nhdt. Trong nghien edu ndy, hgp ehdt quinazoion d y o c t i n g hgp t d ethyl 2-amino-4,5- bis(2-methoxyethoxy)benzoat, ammonium format va formamid. Phan tieh cau true cua san pham 6,7- bis (2-methoxy-ethoxy) quinazoion bang p h i 'H-NMR eho cac tfn hipu t y a n g dng vdi dp djeh chuyen hod hpe eua cac proton tren cau true eiia quinazoion nhy trong nhidu bai bao '''"^'. Pho MS cung x u l t hidn cac pic the hipn khdi lygng ion phan manh dge tryng t d hap chat quinazoion: [M+Hj* = 295,1; IM+2H]+= 296,3; [M+Na]*= 317,1 ;vaMLT=
294,2. Trdn eo s d cac k i t qua pho thu d y g c , chiing tdi cd the k i t ludn rang hgp c h i t 6,7-bis(2- methoxyethoxy) quinazoion da d y g c tong hgp thanh cdng tren co s d phan dng ddng vdng vdi ammonium format va formamid.
Nhdm khao sat anh hydng cua nhipt dp d i n hieu s u i t eua phan dng, phan dng d y a c t i l n hanh khao sat d cac nhiet dd 130, 140, 150 va 160 "C. Hdn hgp sau phan dng d y g c x d ly t y a n g t y nhau, hidu s u i t d y g c xae dmh dya vao khdi lygng san pham sau khi d y g e k i t tinh lai.
Phan dng d y g c t i l n hanh d 130, 140 vd 150 "C va theo ddi t i l n trinh phan dng bang sac ky Idp mdng sau tdng 60 phut Sau 05 gid, khdng nhan thay cd s y thay doi dang k l ve thanh phan phan yng. T i l p tye phan dng qua ddm va theo ddi sau 24 gid. Phan dng d y g e thyc hidn lai d nhiet do 150 "C va cung d y g e theo doi bdng s i c k^ Idp mdng, sau 05 gid nhdn thay cd vet san pham trdn ban mdng sdc ky. T i l p tyc thye hien phan dng d i n 24 gid. Tiep tuc thyc hidn lai phan dng d 160 °C, phan dng d y g e theo doi bdng sac ky ldp mdng moi gid va nhan thdy kdt thue sau 02 gid vdi vdt ethyl 2-amino-4,5-bis(2-methoxyethoxy) benzoat khdng cdn hidn dien tren ban mdng. Sau 02 gid hdn hgp phan dng d y g c x d ly va thu d y g c san pham vdi hidu s u i t 87% (hinh 2).
T^P CHi DlTQC HQC - 7/2015 (SO 471 NAM 55)
47• Nghien ci>u - Ky thuat
=m
NUetdo(*C}Hinh 2; BiSu do kdt qua khao sat nhiet do phan wng
Anh hydng eiia nhipt dp d i n hieu s u i t ciia phdn dng rat Idn do phan dng xdy ra theo ea e h l cOa phdn dng Leuckart. Phan dng b i t dau vdi s y phdn huy eua ammonium format thanh acid formic va amoniae, s y phan huy ndy can thu mgt nang lygng Idn d l b i t dau phan dng. Ngoai ra, phdn dng Leuckart la mpt phdn dng dge thu eiia phan dng egng nueleophiiic vao noi ddi earbonyl, trong dd, giai doan tae nhan nueleophii tac kich vdo tdm thieu dipn t d cung cdn cd mdt nang lygng kieh hogt d l b i t dau xay ra i"i.
San pham 4-chloro-6,7-hls-(2^ethoxyethoxy) quinazolin (5)
Trong nghidn cdu nay, chimg tdi nghidn cdu s d dyng tae chat oxalyl chlorid d l chlor hod hgp chat 4-ehloro-6,7-bis-(2-methoxyethoxy) quinazolin. Nhieu de tdi nghidn cdu gan day cho thay day Id phyang phap an todn va hieu qua trong tong hap hod d y g e . K i t qua pho proton
eua hgp ehat 4-ehloro-6,7-bis-(2-methoxyethoxy) quinazolin eho thay ty lp dipn tich pie eua cdc proton dgc tryng x u l t hien tren pho t y a n g dng vdi ty le so proton cua cac proton nay hi^n di^n trdn cau trtie cua hgp ehat 4-chloro-6,7-bis-(2- methoxyethoxy) quinazolin. Pho MS do trong methanol x u l t hien eae p c ttie hipn ktioi lygng ion phan manh dge trung cua san p h i m 4-ehloro-6,7- bis-(2-methoxy-ethoxy)-quinazolin: [M(^*CI )+H]*
= 313,1; [M("CI )+H]-= 315.1; [M(35CI )+2H]- = 314,3; [M(3'CI )+2H]* = 316,1; [M(35Ci )+Na]* = 335,1; vd [M(3'CI )+Na] = 337,1. Chiing toi khang djnh da t i n g h g p thanh edng sdn pham 4-chloro- 6,7-bis-(2-methoxyethoxy) quinazolin.
San pham erlotinib h y d r o c h l o r i d (6) Phan dng tong hgp eriotinib t d d i n xult 4-ehloroquinazolin d y g c thyc hipn dya trdn phan dng t h l di nhan tham, trong dd tde nhdn nueleophii s d dyng la 4-ethynylanilin 3e tao thdnh san pham eriotinib hydrochlorid vdi hipu suat eao.
De nang eao dp tinh khiet sdn pham d y g c k i t Hnh 2 l l n trong MeOH thu d y g c erlotinib dp tinh khilt 98,2%. Tren pho 'H-NMR (hinh 3) thu d y g c cho thay cae tfn hieu proton dge tm-ng eiia sdn phim eriotinib. Cy t h l cac proton ciia vdng quinazolin va ciia 3-ethynylanilin (H^^°) xuat hi$n d trydng thap, vung eac tfn hidu cua nhdm tham dge tryng:
8,70 (s, I H , HAr); 8,14 (s,1H, H-Ar); 7,70-7,81 (m, 2H, H-Ar); 7,28 - 7,51 (m, 3H, HAr); cdc tin hipu d viing khoang 4 ppm ciia cac carbon mang oxygen (H1-3 1^1-3^. ni5t tjn hipu eua nhdm methyn d 3,67 ppm (s, I H , C = CjH)- So sdnh vdi tdi lipu tham khdo da xdc nhgn san pham Id eriotinib '*^.
1 Writ L L B
Hinh 3: Ph6 'H-NMR cua eriotinib.HCl
TAP CHf Db'gIC Hpc - 7/2015 (S6 471 NAM 55)
• Nghien CLFU - Ky thuat
Ket qua khdi p h i eua san p h i m (hinh 4) eung cho thay cac pie dge tryng eua eriotinib.
Cdc pic HRMS (ESi-MS). [M{^^C\)Y = 428,0;
[M(3'CI)]+ = 430,9; M^^= 429,9. T d ede d d lieu pho trdn, khang djnh san pham eriotinib d y g e tong hgp thdnh cdng vdi hipu s u i t va dp tinh khilt eao.
Hinh 4: Pho MS cua eriotinib hydrochlorid
Ket luan
Trong nghidn c d u nay, tren ea s d ede k i t qud phan tfeh eau true eho t h i y eriotinib hydrochlorid dyge t i n g hgp todn phan thanh edng qua quy trinh 6 phan dng. Cae phan dng d y g c lya ehpn d i l u kipn phan dng vd tdc chat thich hgp d l phdn dng CO hipu qua kinh t l . K i t qua hieu s u i t tong hgp todn phan tren 42% vd dp tinh khilt tren 98%. Hydng nghien cdu hien dang t i l p tyc d y g c edi t i l n d l ndng cao hipu qua eua todn quy trinh, t i l n den nghien edu tong h g p quy md pilot va tgo t i l n d l san xuat eriotinib hydrochlorid lam nguydn lipu cho thude ehdng ung t h y a Vipt Nam.
LOT cam on: Cdc tdc gia chdn thdnh cam an Sa Khoa hgc Cdng nghe Tp. Ho Chl hJlinh da tai tr^kinh phi thwc hien de tdi.
Tai lieu tham khao
1 Grands, J. R., Sok, J. C. (2004). "Signaling through the epidermal growth factor receptor during the development of malignancy", Pharmacol. Ther..
37,102.
2. Chandregowda V. Kush AK, Chandrasekara Reddy G, (2009), "Synthesis and in vitro antitumor activities of novel 4-anilinoquinazoltn derivatives", European Joumal of Medicinal Chemistry. 44(7), 3046-3055.
3. Rodney Caughren Schnur, Lee Daniel Amol (1998), "alkynyl and azido-substituted 4-anilinoquinazolins", United Stases Patent, Appl, Nc 653, 786
4. Venkateshappa Chandregowda, Gudapati Venkateswara Rao, Goukanapalli Chandrasekara Reddy (2007), "Convergent approach for commercial of gefitinib and erlolmib", Organic Process Research &
Development. 11, pp. 813-816
5. Petr KnesI, Dirk Roseling, Ulrich Jordis. (2006),
"Improved synthesis of substituted 6,7-dihydroxy-4- quinazolinamms: tandutinib, erlotinib and gefitinib", Mo/ecu/es, 11, pp 286-297.
6. Leila Barghi, Ayuob Aghanejad, Hadi Valizadeh, Jaleh Barar, Davoud Asgan (2012): "Modified synthesis of erlotinib hydrochlorid", Advanced Pharmaceutical Bulletin, 2{^).pp^^9-•\22.
7 Gengzhen Zhang, Linlin Zha (2012), "Isolation of highly pure erlotinib hydrochlorid by recrystallization after nueleophiiic substitution of an impunty with piperazine". Res. Chem Intermed, Doi 101007/
S11164-012-0757-9
8. Vietnamese Oncology Association (2013), Oncology Symposium on Lung and Liver Cancer, Hanoi, Vietnam.
9. EunyoungByun, BomiHong.KattiliaA. DeCastro, Minkyung Lim, and Hakjune Rhee (2007),
"One-pot reductive mono-n-alkylation of aniline and nitroarene derivatives using aldehydes". J. Org.
Cftem. .72,9815-9817
10 Dan Wang, Feng Gao (2013), "Quinazolin derivatives: synthesis and bioactivities", Chemistry Central Journal, 7, 95.
11. Elliot R. Alexander, Ruth Bowman Wildman (1948), "Studies on the mechanism of the leuckart reaction", J. Am. Chem Soc , 70 (3), pp 1187-1189 {Ngay nhan bdi: 28/01/2015 - Ngay duyet dang: 02/07/2015)
