TAP CHI HOA HOC
DOL 10.15625/0866-7144.2015-0084
T.53(1) 35-40 THANG 2 NAM 2015
NGHIEN CLPU T 6 N G HOP CHON LOC 10j8-AZIDOARTEMISININ
Le Huy Binh, Nguyen Thanh Binh, Nguyen Thi Thiiy Van, Tran Khic VG*
Bd mdn Cdng nghe Hda dugc vd Bdo v | Thuc vgt, Vien Ky thugt Hda hgc, Trudng Dgi hgc Bdch khoa Hd Ngi
Din Tda soan 7-7-2014; Chip nhan dang 4-12-2014
Abstract
This paper presents a simple method for selective synthesis of the key compound, 10/?-azidoartemisinin. The compound lOyS-azidoartemisinin was selectively synthesized by the reaction of dihydroartemisinin (2) with NaN], KI using ZnCl2 as a catalyst at 0-5 °C. The strucmre of 10^-azidoartemisinin was elucidated based on 'H, '^CNMR spectmm and known documents.
Keywords. Artemisinin, dihydroartemisinin, artemether, arteether, sodium azide.
l.DATVANDE
Artemisinin (1) li mgt sesquitecpen lacton chiet xuat tir cay thanh hao hoa vang (Artemisia annua L.) dugc sii dyng lam nguyen lieu dau quan trgng eho eac nghien ciru kham phi thude sdt ret [1]. Mdt sd hgp chat ban tdng hgp cd hogt tinh khing sdt ret nhu: artemether (3a), arteether (3b) va artesunat (3c) (hinh 1) dugc ban tong hgp tir dihydroartemisinin (2) hi?n dang dugc su dyng trong dilu fri lim sang sdt ret [2-5]. Gan day, cac thii ngjiiem lim sang edn cho thay nhieu dan xuit ciia artemisinin the hien mot sd hoat tinh sinh hgc khac nhu: boat tinh khang u, khang virus, khang nam va lic che mien dich [6].
Trong sd cac din xuit cua artemisinin dugc tdng hgp gin day dang chii y la cac din xuit chira nhan triazole the d vi fri C-10 cua khung artemisinin. Cac dan xuit nay dugc tdng hgp qua phan iing cdng ddng vdng 1,3 giira hgp chat frung gian 10- azidoartemisinin vdi cac ankyn. Mdt sd dan xuat kilu nay the hien hogt tinh irc chi tl bao ung thu vii mgnh hgn ca Taxol [7].
Cac dan xuat cua artemisinin chira nito d vi fri C-10 dugc Haynes va cgng sy [8, 9] dieu che lan dau tien qua phin iing cua dihydroartemisinin (2) vdi trimethylsilyl chloride va sodium bromide trong dung mdi toluene d 0 °C cho san pham trung gian lO^-bromodeoxoartemisinin. Chit nay sau dd phan ling ngay vdi cac amine bgc 1 hoac bac 2 dl cho san pham la cac 10-aminoartemisinin. Tren co sd phugng phap nay, Sungsik Cho va cdng su da dieu chi hgp chat lO-azidoartemisinin qua phan irng cua dihydroartemisinin (2) vdi frimethylsiiyi bromide va sodium azide frong dung mdi diclometan d nhiet do
phdng frong 12 gid. San phim thu dugc la hdn hgp ciia 10^-azidoartemisinin (4), 10«-azidoartemisinin (5) va san pham 9a,10^-azidoartemisinin (6) vdi hieu suit lin lugt la 17 %, 21 % va 49 % [7]. Dl nang eao hieu suit cua sin phim, Lijun va cdng si^
dS cai tien quy trinh bang each sir dyng trimethylsilyl bromide va sodium azide vdi su cd mat Clia mgt lugng nhd KI (2,5 % moi) trong dung mdi diclometan d nhiet dd phdng trong 28 gid thu dugc san pham 10^-azidoartemisinin (4) vdi hieu suit 47% va mdt lugng nhd san pham lOa- azidoartemisinin (5%) [10]. Theo hiiu biit ciia chiing tdi, hien chua cd bao cao ve viec dieu che cic lO-azidoartemisinin sir dung kem clorua lam chit xiic tie. Trong qua trinh nghien ciiu tdng hgp cac din xuat mdi cua artemisinin va sang Igc hogt tinh khang ung thu, chiing tdi phai su dyng chit chia khda lO^-azidoartemisinin. Vi vay, bai nay chiing tdi frinh bay tdng hgp chgn Igc hgp chit lO^-azidoartemisinin sir dung kem clorua lam chit xiic tac.
2. THl/C NGHIEM
Dihydroartemisinin mua tir cdng ty Dugc khoa, Trudng Dgi hgc Dugc Ha Ngi. Cic tac nhin va dung mdi phan iing mua ciia hang Merck va Aldrich.
Diem ehay dugc do fren may Electrothermal IA 9200 Shimadzu. Phd ' H NMR va '^C NMR dugc do tren may Bruker AVANCE 500 MHz tgi Vien Hda hgc, sii dyng dung mdi do CDCI3. Do chuyln dich hda hgc (5) tinh bing ppm so vdi chit chuin (TMS).
Hing so tuong tic (J) dugc bilu diln bing Hz. Tiln trinh phan iing dugc theo ddi bdi sic ky Idp mdng
TCHH, T. 53(1), 2015
(TLC) sir dyng ban nhdm frang san (Merck 60 F254). sic 1^ cgt sir dung silica gel cd hgt 40-230
Trdn Khdc VU vd cdng sir mesh.
Dihydroartemisinin (2)
a. R = CHj artemetiier b. R = CH1CH3 arteother c. R = COCH2CH2COONa artesunats
Hinh 1: Mdt so dan xuit ciia artemisinin 2.1, Quy trinh thuc nghiem tong hgp
1 Op-azidoartem isinin
Mdt hdn hgp cua dihydroartemisinin (1100 mg, 3,87 mmol), NaNj (772 mg, 5,81 mmol, 1,5 eq), ZnCb (377 mg, 5,81 mmol, 1,5 eq), KI (16 mg, 0,097 mmol, 0,025 eq) frong diclometan khan (15 mL) dugc lam lanh tdi 0-5 °C bang nude di. Phan ling dugc khuay d nhi?t do nay frong 8 gid. Kiem tra phan ling bang sac ky Idp mdng sir dung he dung mdi friln khai:n-hexan:etyl axetat (9,5:0,5). Sau khi phin ling ket thiic, them vao hdn hgp phan iing diclometan (20 mL) va hdn hgp phin iing dugc chiet vdi nude (3x15 mL). Pha hiru co dugc tach ra va lam khan vdi Na2S04 khan. Pha hiiu co sau dd duge cit loai dung mdi dudi ap suit giam. Can phan irng dugc tich bang sac ky cgt silica gel sir dung he dung mdi «-hexan:etyl axetat (95:5) thu dugc san pham 10^-azidoartemisinin (334 mg, 28 %). Tinh the miu tring: dne; 41-43 °C. ' H NMR (500 MHz, CDCI3) 5,53 (s, IH, H-12); 5,37 (d,J=4,0 Hz, IH, H-10):
2,71 (m, IH); 2,40-2,33 (m, IH); 2,06-2,03 (m, IH):
1,91-1,86 (m, IH); 1,82-1,81 (m, IH); 1,89-1,87 (m, IH); 1,82-1,76 (m, IH); 1,72-1,63 (m, 2H); 1,52- 1,47 (m, 2H); 1,44-1,42 (m, 3H); 1,37-1,34 (m, IH);
1,26-1,22 (m, IH); 0,96-0,90 (m, 6 H). '^C NMR (125 MHz, CDCI3) 5: 104,4 (C-12); 91,8 (C-3); 88,6 (C-12a); 80,6 (C-10); 52,5; 44,1; 37,3; 36,2; 34,5;
30,2; 25,9; 24,6; 23,5 (C-14); 20,3 (C-15); 13,1 (C-16).
Sir dyng xiic tic (CH3)3SiCl theo quy frinh d bang 3 thu dugc lOa-azidoartemisinin (15 %). Tinh thi mau tring: dne: 101-103 °C. ' H NMR (500 MHz, CDCI3) 5: 5,31 (s, IH, H-12); 4,54 (d, J= 10 Hz, IH, H-10); 2,37-2,28 (m, 2H); 1,99-1,95 (m, IH); 1,87-1,80 (m, IH); 1,71-1,63 (m, 2H); 1,71- 1,63 (m, IH); 1,54-1,49 (m, IH); 1,45-1,39 (m, IH);
1,37 (s, 3H, H-14); 1,28-1,17 (m, 3H); 0,98-0,94 (m, l H ) ; 0 , 8 9 ( d , / = 6 , 0 H z , 3H, H - 1 5 ) ; 0 , 8 5 ( d , / = 7 Hz, 3H, H-16). '^C NMR (125 MHz, CDCI3) 5:
104,7 (C-12); 91,7 (C-3); 87,7 (C-12a); 79,9 (C-10);
51,5; 45,2; 37,3; 36,1; 34,0; 32,4; 25,8; 24,6; 21,7 (C-14); 20,2 (C-15); 12,8 (C-16).
3. KET QUA VA T H A O LUAN
Cac din xuit 10-azidoartemisinin la hgp chat chia khda rit quan trgng dl tdng hgp cic din xuat mdi Clia artemisinin chiia nhan triazole sir dyng phan irng Click. Gin day mgt sd dan xuat kiSu nay da dugc tdng hgp va danh gii hogt tinh khing vdi mot sd ddng t l bao ung thu. Kit qua thu nghiem cho thay mgt sd din xuat kilu nay cd hoat tinh rit mgnh ngang so vdi mgt sd thude fri ung thu dang dung nhu Cispaltin hay Taxol [7]. Tuy nhien, nguyen lieu dau su dyng la cac hgp chat lOy? va lOa-azido- artemisinin, ehii yeu la lO^-azidoartemisinin can lugng ldn de cho cac chuyen hda tiep theo. Vi vay, chiing tdi dat vin d l tdi uu cac dieu kien dl thu dugc cac hgp chit nay mgt each chgn loc, thuan tien hon cho qua trinh tich sin phim tir hdn hgp phan iing.
Qua frinh tdng hgp 10-azidoartemisinin dugc minh hga nhu 0 so do 1.
Theo so dd tdng hgp chung d fren, nguyen lieu de dieu che cac dan xuat 10-azidoartemisinin di tir hgp chat 2. Hgp chat nay phan irng vdi cac tac nhan nucleophile (N3") nhu NaN3, hoac (CH3)3SiN3 sii dyng cac axit Lewis khac nhau nhu (CH3)3SiCl hoac (CH3)3SiBr d cac dilu kifn khac nhau [7-10]. San phim thu dugc gdm: lOyff-azidoartemisinin (4), lOa-azidoartemisinin (5) va sin phim dao ciu hinh d C-9 (6). Ve mat ca chi, phan iing xay ra theo co che SNI qua su hinh thanh ciia oxonium cacbocation.
Diy la budc xay ra nhanh dudi tac dyng cua xue tac la axit Lewis. Giai dogn tilp theo la sir tin cdng ciia cac nucleophile vao ca hai phia ciia oxonium cacbocation va cho san phim cd ciu hinh B (7) va a (8). Phan iing cdn bj anh hudng ciia cac yeu td nhi^
ddng lien quan din su bin virng ciia oxonium cacbocation, nen de bi mit nude dl tgo thanh san pham la 9,10-dehydroartemisinin (10) va san phim dao ciu hinh 11 (hinh 2).
36
TCHH, T. 53(1), 2015 Nghien cuu tdng hap chgn Igc.
Sa do 1: Tdng hgp cic dan xuit 10-azidoartemisinin
Hinh 2. Co che phan iing cua DFIA vdi NaN3 sir dyng xiic tic axit Lewis Nhu vgy, mac dii phin iing xay ra theo co che
phin img the SNI, giai dogn cac nucleophile tan cdng vao oxonium cacbocation mdi la giai dogn quyet dinh. Tuy nhien, frong phin iing nay, yeu td nhiet dgng va xuc tic cung anh hudng nhieu den do chgn Igc ciing nhu hieu suit chung ciia sin pham 10-azidoartemisinin thu dugc. Chinh vi vay chung tdi dgt vin de khao sat cic dieu kien de thu dugc sin phim azido chgn Igc phyc vy cho cac bien ddi hda
hgc tiep theo. De thu dugc san pham chgn Igc 10^-azidoartemlsinin (4), chung tdi khao sat mdt so yeu td inh hudng den do chgn loc cua san phim tren CO sd nghien ciiu ciia Lijun va cgng su [10]. Trong nghien ciiu nay, chiing tdi sit dung ty le tac nhan NaN3 va ddng xiic tac KI nhu trong tai lieu, chi thay ddi logi axit Lewis va nhiet do cua phan iing. Tham khio cic tai ll?u Hen quan, chiing tdi thiy ring ZnCL la mdt axit L^wls rit em diu nhung chua dugc
TCHH, T. 53(1), 2015
suf dyng trong phan irng de dieu che 10- azidoartemisinin. Chiing toi suy doan rSng, su em diu Clia axit Lewis k a hap voi nhiet dp CO the anh huong d6n su bSn vung cua oxonium cacbocation, tu do se anh huong den do chpn Ipc ciia phan ung.
D6 khao sat, thoi gian phan ung dupe chpn la 8 gio ii CO the xu ly trong ngay lam viec. Kiem tra phan img bing sac ky lop mong he dung moi trien khai n-hexan:etyl axetat (9,5:0,5). Trong tit ca cac
Trdn Khdc VU va CQngs^
Uuong hpp dSu xuSt hien san phim dao cau hinh 6c6 Rf \&n nhit, sau do la san phim ciu hinh ^ ( 4 ) va a (5). Tuy nhien, san phim dao cau hinh xuat hien a lupng vSt Uen ban mong nen chi quan tam hai san phim chinh CO ciu hinh fi (4) va a (5). Qua Uinh khao sat cac diSu kien tang hop chpii Ipc 10^- azidpartemisinindupc mo ta nhu o so do 2 va cac bang duoi day.
CHs CH3 CH3
H s C ^ O O
O 10 • OH
H3C-'° y
H s C ^ ONaNj, CH2CI2, Kl 8h, rt
°H'
N3 Sadd 2: T6ng hpp chpn Ipc lOP-azidoartemisinin Bang 1: T6ng hpp 1 OP-azidpartemisinin 6 nhi?t dp phong trong 8 gicj
TT 1 2 3 4 5 6
Xiic tac (CH3))SiCI (CH3)3SiCI (CH3)3SiCI ZnCI;
ZnCl2 ZnCh
DHA, mg 1030
980 870 1200 1150 775
Tyie (DHA/axit/NaNj/KI)
1/1,5/1,5/0,025 1/1,5/1,5/0,025 1/1,5/1,5/0,025 1/1,5/1,5/0,025 1/1,5/1,5/0,025 1/1,5/1,5/0,025
San phim 10,8, %
75 72 76 65 69 66
San pham 10a, %
20 23 21 30 27 25
San pham dao cau hinh, %
Vet nho Vet nho Vet nho Vetnho Vet nho Vet nho Mdi phan iing dugc thyc hien 3 lin. Hieu suit khdng uu tien dl cho chgn Igc sin pham ca cau hinh phan ling dugc tinh la trung binh cua 3 lan thi P va a. Chung tdi tilp tyc khao sat d nhiet dp da nghiem. Theo kit qua khio sit d bang fren,d nhiet mudi (am 10 din im 5 °C). Kit qua dugc trinh bay dg thudng ca xiic tic ZnC^ va (CH3)3SiCl deu frong bang 2.
Bdng 2: Tdng hgp lOp-azidoartemisinln d nhl^t do da mudi trong 8 gid TT
1 2 3 4 5 6
Tac nhan (CH3)3SiCI (CH3)3SiCI (CH3)3SiCI ZnCI;
ZnCb ZnCl2
DHA, mg 1100
970 870 205 175 170
Tyle {DHA/Axit/NaN3/ICI)
1/1,5/1,5/0,025 1/1,5/1,5/0,025 1/1,5/1,5/0,025 1/1,5/1,5/0,025 1/1,5/1,5/0,025 1/1,5/1,5/0,025
San pham 10^,%
65 56 58 V6t V6t
va
San phim 10a, %
20 12 12 0 0 0
San pham dao cau hinh, %
Vet Vet
va
0 0 0 K a qua o bang 2 cho thiy, o nhiet dp da mu6i xiic tac la ZnCb phan img hiu nhu khong xay ra. (J phan ling chi xay ra voi xuc tac la (CH3)3SiCl. Voi nhiet dp nay kha nang xdc tac ciia ZnCbtro nen mat
TCHH, T. 53(1), 2015
tac dyng vi khdng hinh thanh dugc oxonium cacbocation. Vi vay, cung khdng xuit hien sin phim dao cau hinh nhu thudng thiy khi su dung axit Lewis manh nhu: (CH3)3SiCI, Sau 8 gid, phan iing
Nghien cuu tdng hgp chgn Igc...
chi xay ra vdi lugng vit ciia san phim 10^.Vi viy, phan img duoc tilp tuc khao sat d nhiet do nude di (0-5''C).
Bdng 3: Tdng hgp 1 Op-azidoartemisinin d nhiet do nude da trong 8 gid TT
1 2 3 4 5 6
Tac nhan (CHj)3SiCI (CH3)3SiCI (CH3)3SiCI ZnCl2 ZnCli ZnCI;
DHA, mg 1100
970 870 205 175 170
Tyl$
(DHA/Axit/NaN3/Kl) 1/1,5/1,5/0,025 1/1,5/1,5/0,025 1/1,5/1,5/0,025 1/1,5/1,5/0,025 1/1,5/1,5/0,025 1/1,5/1,5/0,025
San pham lOjS, %
80 75 77 35 33 28
San pham lOa. %
15 18 16 0 0 0
San pham dao cau hinh, %
va
Vet Vet 0 0 0 K6t qui cho thiyd nhiet dg nude da phan iing
xay ra vdi hieu suit ehuyen hda kha cao va khi chgn loc khi sii dyng axit Lewis la (CH3)3SiCl. 6 nhiet do nay khi sir dyng axit Lewis la ZnC^thu dugc san phim chgn Igc lO^-azidoartemlsinin mac dii hieu suit chi khoang 28-35 %, thip hon nhilu so vdi khi dung axlt Lewis la (CH3)3SiCl. Mgc dii vgy, diy cung la lan dau tien ZnCl2 dugc sir dyng cho kieu phin ling nay va cho chgn Igc lap the. Vdi ket qua nay, khi nang quy md phan ung len co gram sii dyng ZnCl2 vin cho hieu suat tuong duong nhu tren. Nhu viy, diy la ket qua ban dau lam nen tang de cd the toi uu them cac dieu kien khac de cd the thu dugc san pham 10^-azidoartemisinin vdi hieu suit cao hon. c i u tnic cua san phim 10^-azidoartemlslnin va lOa-azidoartemisinin thu dugc tir cic thi nghiem tren dugc xac dinh sii dung phd ' H va '^C NMR va so sanh vdi cac tai lieu da cdng bd [10].
4. KET LUAN
Lin diu tien ZnCl2 dugc sir dyng nhu la mdt xuc tie em dju, de kiem va tuong ddi re tien cho phan iirng dieu che chgn loc 10;3-azidoartemisinin, mdt hgp chit chia khda quan frgng cho phan ling dieu che cac dan xuit friazole cua artemisinin.
IM cam gn. Cdng trinh ndy dugc hodn thdnh vdi su tdi trg' cua Quy khoa hgc cdng nghi qudc gia (NAFOSTED) qua de tdi. ma sd 104.01-2013.01.
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Lien he: Tran K h i c V u
Bg mdn C d n g nghe Hda d u g c va B a o v e T h u c vat Vien Ky thugt Hda h g c , T r u d n g Dai hgc Bach klioa H i N g i Sd 1, Dgi Cd Viet, Hai Ba T r u n g , Ha N g i , Viet N a m E-mail: [email protected]
Dien thogi: 0 9 0 4 3 0 6 9 2 5 .
