TAP CHI HOA HOC
DOI: 10.I5625/0866-7144.2014-0009
T. 52(4) 443^146 THANG 8 NAM 2014
T 6 N G Hgp 21-ACETAT PREGNA9{11)-DIEN-17a,21-DIOL-3,20-DION
Lini Dire Huy', Nguyen Thj Olep, Pham Thanh Thao, Lun Due Hung Phong thi nghidm Hoa hgc Steroid vd Alkaloid, Vi^n Hda hgc,
Vi$n Hdn ldm Khoa hgc vd Cong nghi Vi^t Nam Den Tda sogn I3-0I-20I4
Abstract
in this paper, we present the synthesis of the 21-acetate pregna-4,9(ll)-diene-17a,21-diol-3,20-dione from 9a- hydroxy androstenedione, an important intermediate for synthesis of corticoids such as hydrocortisone and prednisolone. The structure of the product and its intermediates vi'as examined by physical methods such as: IR, MS, 'H and "C NMR.
Keywords: 21-acetate pregna-4,9(ll)-diene-17a,21-diol-3,20-dione,9a-hydroxy androstenedione.
1. u6
DAUNghien cim chuyen hda vi sinh hdn hgp sterol den cac hgp chat 17-ceto steroid trung gian nhu androstendion (AD), androstadiendion (ADD), 9a- hydroxy androstendion (9a-0H AD) dang phat trien mgnh me [1, 2]. Chung tdi cung da cdng bo phuang phap hipu qua chiet xuat tmh che phytosterol tir phy thai cdng nghipp tinh luypn dau dgu tuang [3, 4] va chuyen hda vi sinh chiing den AD, ADD va 9a-0H AD [5-9]. Da cd nhieu cdng trinh nghian cuu tong hgp cocticoid tir 17-cetosteroid tir 17-ceto steroid [10- 16]. Chung tdi da cdng bo ket qua nghien curu mgt s6 phuang phap tgng hgp cocticoid tir 17-ceto steroid [17-27]. Tiep tyc hudng nghien cihi nay, trong bai bao nay chiing tdi trinh bay mgt phuang an tong bgp 21 -acetat pregn-4,9( 11 )-dien-17a,21 -diol-3,20-dion tir 9a-hydroxy androstendion, mgt hgp chat trung gian quan trgng de tong hgp cocticoid nhu la hydrococtison va prednisolon.
2.THUCNGH1$M
Phan chung Diem chay dugc do tren may Boetius (CHDC Due). Pho hong ngogi (IR) ghi tran miy FT-IR-IMPACT^IO dgng vien nen trong KBr.
Pho cdng hudng tir hgt nhan dugc ghi tran may Bruker 500 MHz, chat chuan TMS, dung mdi CDCI3.
Dung mdi hda chit mua ciia Cty Merck va Aldrich.
Kiem tra djnh ti'nh diing ban mdng de nhdm trang sin Merck 6OF254. Pho khoi lugng (MS) dugc ghi tran mAy Agilent 6310 Ion Trap, dung mdi CHCI3.
Androst-4,9(n)-dien-3,17-dion H. Hgp chit II dugc tdng hgp theo phuang phap rieng ciia chung tdi
da dugc md ta trong [28], Dc. ("C): 201-203°. Hipu suit 95 %. Pho IR (y, cm"', KBr): 1732 va 1659 (C=0 d cac vi tri 17 va 3) C=C d 1616 va 1449 (C=C). Phd 'H-NMR (CDCI3, ppm, 500 MHz): 5,76 (br s, IH, H-4); 5,57 (t, IH, H-11); 1,36 (s, 3H, CH3- 19);0,88(s,3H,CH3-I8).
17a-hydroxy-17p-cyanoandrost-4,9(ll)-dien- 3-on n i . Tham 100 ml axeton cyanbidrin va 10 ml dung dich NaOH 3,3 M (pH - 8) vao hdn hgp gdm 100 g bgp chit n trong 250 ml MeOH. Khuay dao hdn hgp phan img trong I gid d 35 °C. Nhd gigt 60 ml H2O cat vao hdn hgp phan ung. De yen qua dem, sau dd them tiep 200 ml H2O cat va khuay dao 2 gid d nhipt dg phdng. Lgc, rira hdn bprp phan ung blng bang H2O den pH = 7. Dd bd djch lpc, tiia tren pheu dem say khd trong tii say chan khdng d 45 °C, nhgn dugc 87,2 g san pham IU (Hipu suat 80,0 %). Dc.
("C): 182-183 (EtOAc). Pho IR (y, cm', KBr): 3236 (0-H); 1645 (C=0 d vj tri 3); 1528 va 1452 (C=C).
Pho MS: 311 [M"]; 284 [M-HCN]^ Pho 'H-NMR (CDCI3, ppm, 500MHz): 5,76 (br s, IH, H-4), 5,57 (d, IH, H-U); 2,97 (s, IH, OH-17); 1,34 (s, 3H, CH3-I9); 0,93 ppm (s, 3H, CH3-I8).
17a-hydroxy-3,3 (2,2-dinietylpropylendioxy)- androst-5,9(ll)-dien-17p-cacbonitril IV. Hda tan 84 g hgp chit III trong 300 ml CH2CI2. Them vao dung djch 120 g neo-pentyl glycol, 8,0 g p-TSA va 80 ml trietyl octofocmat. Cho N2 khan qua hp phan ling. Khuay dao hdn hgp phan img d 0^-H5 °C. Sau 8 gid dimg phan img, trung hda blng 8,0 ml trietyl amin. Them vao 700 ml H2O, da yen qua dem. Tiia dugc tach ra, lgc va rira chat rln nhgn dugc blng H2O dan pH = 7. Say khd chat rln la san pham, nhgn dugc 105,1 g IV (hipu suit 98 %). Dc.(°C): 228-231
443,
TCHH, T. 52(4), 2014
(axeton). Pho IR (y, c m ' , KBr): 3423 (O-H), 1673 va 1641 (C=C); 1105 (ketal). Phd 'H-NMR (CDCI3, ppm, 500 MHz): 5,54 (d, IH, H-I I); 5,42 (d, 1 ^ H- 6); 3,55-3,45 (m, 4H, H-ketal); 1,19 (s, 3H, CH3-I9);
0,99 (s, 3H, CH3-I8); 0,93 (s, 6H, 2 nhdm CH3- ketal).
Pregn-4,9(ll)-dien-17o-ol-3,20-dion VI. Hda tan 80 g san phim IV trong 700 ml dietyl ete trong ddng N2 khan ddng thdi khuiy dao d nhiet dg phdng.
Tham vao 80 ml vinyl etyl ete. Lam lanh hdn hgp phan ling xudng -5-H-5 °C va them vao 8,0 g p-TSA, ddng thdi khuiy dao trong ddng N2 khan. Sau 4 gid them vao hp phan ung 8,0 ml EtsN. Hg nhipt dg phan ling xudng -20 °C dd tham vao 350 ml CHsLi 1,6 M trong Et20. Nhd gigt CHaLi xong thi dua he phan ling ra ngoai nhipt do phdng cho den khi phan umg dat nhipt dg phdng thi tham vao hdn hgp MeOH;
HzOiHCl (d) = 40 ml:2 ml:2 ml. Tidp tuc nhd gipt hdn hgp H2O : HCl (d) = 80 ml:80 ml. Chung clt lly Et20 tren may khuay tir gia nhiet sao cho nhiet dp trong binh phan u'ng < 60 °C. Khi trong he phan ung xuat hipn nhQ tuong thi nhd gigt vao he phan img 1200 ml H2O. De d ngan dudi tu lanh. Lgc, rua hdn hgp phan ung bang H2O (140 ml x 5) dan pH = 7.
Dich Igc do bd, tiia dem say khd trong may say chan khdng. Nhan dugc 74,0 g san pham thd. Hda tan san pham thd trong 1500 ml CH2CI2, them vao 15 g than hogt tinh, khuay dao d nhipt dp phdng trong 1 gid.
Lpc, rua bang CH2CI2 (150 ml x 2). Chung cat dich Igc dong thdi khuay dao tren may khuay tir gia nhipt dan the tich 150 ml, them vao 220 ml axeton, cat tiap dan cdn ~ 140 ml. De ngan dudi tii lanh qua dem. Lgc, rira bang axeton (40 ml x 3), Et20 (40 ml X 2). Can dem sly kho. Cat quay kiet djch lgc rdi them vao 60 ml Et20, khuiy d nhiet dp phdng 45 phiit, de ngan dudi tii Ignh qua dem, Ipc rira blng Et20 (20 ml X 3). Dd bd djch Igc, can say khd trong til say chan khong. Nhgn dugc 61,8 g (hieu suit 93
%). Dc. (°C): 215-217 (axeton). Phd IR (7, cm"', KBr): 3475 (O-H), 1707 va 1660 ( C - 0 d vi tri 20 va 3), 1621 va 1443 (C=C). Phd ' H - N M R (CDCI3, ppm, 500 MHz): 5,74 (s, IH, H-4); 5,54 (t, IH, H- II); 2,75 (s, 3H, CH3-2I); 1,34 (s, 3H, CH3-I9);
0,69(s,3H,CH3-I8).
21-axetat pregn-4(9,ll)-dien-17a,21-diol-3^0- dion VIII. Hda tan 67 g VI trong 500 ml CH2CI2 va 150 ml MeOH. Them vao 50 g CaO, 3,4 ml H2O;
khuay dao d nhipt dg phdng trong ddng N2 khan.
Nhd gigt dung djch 10,4 g CaC^ va 68 g I2 trong 155 ml MeOH. Sau 1,5 gid dimg phan ung, them vao 500 ml CH2CI2. Dd hdn hgp phan irng vao dung djch 300 gNaHCOs trong 3500 ml H2O. Khuiy d k , lgc, njra hdn hgp phan ung blng CH2CI2 (100 ml x 3). Bd egn tren phiu. Chiat lay Idp CH2CI2 d dudi, chiet Idp H2O d tren blng CH2Ci2 (400 ml x 2). Dich
Lmt Dlic Huy vd cgng sit chidt CH2CI2 dem cat quay den khd. NhSn dugc 100,2 g san phim tho 21-iod steroid. Hda tan toan bg san phim thd 21 -iod steroid vua nh§n dugc d tren trong 800 ml dimetyl focmamit, them vao 100 g KOAc, khuiy dao d 60-^65 "C trong khoang tiidi gian 2 gid. Lam ngudi hdn hop phan img rdi do vao 1500 mi nudc, ddng thdi khuiy dao. Lgc rua blng H2O (200 ml X 5) ddn pH = 7. Dich lgc dd bd, bia dem sly khd trong tii say chan khdng d 62 °C nhgn dugc 80,802 g san phim thd. Hda tan san pham tiio trong 1500 ml CH2CI2, them vao 20 g C hoat tinh, khuay d nhiet do phdng 1 gid. Lgc, rira bang CH2C12 (150 ml X 2). Dich Igc clt bdt dung mdi dan tiiS tich 140 ml. Them 250 ml axeton, clt tiap d6n thd tich 150 ml. Dd d ngan dudi tu lanh qua dem. Lpc, riia bang aceton (40 ml x 2), blng Et20 (20 ml x 3). Cgn thu dugc dem sly khd trong tii say chan khdng nhgn dugc 66,8 g san phim (hieu suat 85 %). Dc. ("C):
226-228 (axeton/CH2Cl2). Phd IR (y, cm"', KBr):
3311 (til, O-H), 1732 va 1665 (2xC=0). Phd MS:
286 [M*]; 772 2x[M*]; 368 [M-HjO]^ 327 [M- CH3C00]^ Phd 'H-NMR (CDClj, ppm, 500 MHz);
5,73 (s, IH, H-4); 5,54 (d, IH, H-Il); 4,89 va 5,05 (2 d, 2H, CH2-2I, J = 16 Hz); 2,15 (s, 3H, CHj-Ac);
1,32 (s, 3H, CH3-I9); 0,63 (s, 3H, CH3-I8). Phd "C- NMR (CDCI3, ppm, 500 MHz) xult hipn 23 tin hieu cpng hudng tuong irng vdi 23 nguyen tir C trong phan tir, cd do chuyan djch hda hgc phii hgp nhu sau: 205,2 (C=0 d vi tri 20); 199,5 (C=0 d vj tri 3);
170,6 ( C - 0 ciia nhdm axetyl); 169,9 (C olefinic vj tri 5); 144 (C olefinic vi tri 9); 123,9 (C olefinic dvj tri 4); 118,9 (C olefinic d vi tri II); 89,6 (C-17);
67,9 (CH2-2I); 47,9; 46,7; 40,9; 37,6; 34,6; 34,2;
33,8; 32,9; 32,2; 31,9; 26,2; 24,4; 20,5 va 14,3 ppm (tin hipu ciia cac nguyan tii C trong he thong vdng no).
3. KET QUA v A T H A O LUAN
Cac phan iing nghien cuu duac tdm tit trong scr dd 1.
Digu kien phan ung: (I)-»(II): H*/Ref.;
(II)^(III): axeton cyanhydrin; (lil)-^(IV);
neopentyl glicol; (IV)-»(VI): Vinyl etyl ete, metyl liti; (VI)^(VIII): h/CaO/CaCb/MeOH; KOAc.
Tir 9a-0H AD I chung tdi da tong hgp dugc hi;rp chat A -AD n theo phuong phap rieng da dugc mfi ta trong [28] vdi hipu suit rat tot, san phim dii sgch de cd the diing ngay vao phan ung tiep theo mil khdng cln tinh chd. Pho ' H-NMR ciia II: xult hipn 1 triplet til d 5,57 ppm dugc gan cho proton olefinic it vjtri 11.
Da sii dyng bgp chit A^-AD H la san pham trung gian de thyc hipn phan lirng hai giai dogn tgo mgch
TCHH, T. 52(4), 2014
nhanh pregnan. Dau tien la phan irng cgng hpqj nucleophii nhdm nitril chgn lya lap tha va viing vao nhdm cacbonyl d vj tri C-17 siir dyng tac nhan axeton cyanhydrin trong dung mdi CH3OH d 35 °C trong mdi tnrdng kiem de nhan dugc din xult hydroxynitrit III vdi hipu suat tdt. Phd MS cua HI vdi pic ion phan tir [M*] le cd m/z = 311 va pic ion marih djc trung [M-HCN]"*^ cd m/z = 284. Giai dogn thut 2 tiep theo la metyl hda nbdm nitril, trudc do can bao vp nhdm 3-cacbonyl va I7-hydroxyl. Nhdm cacbonyl dugc ketal hda vdi tac nhan neopentyl glycol trong sy cd mat ciia trietyl ete octofocmat va p-toluen sunfonic axit. Dan xuat 3-ketal IV nhan dugc vdi hieu suit tdt va dugc khang dinh cau tnic
Tong hgp 21-aceiatpregn-4.9(11)...
bang cac phuang phap pho. Bao vp nbdm 17- bydroxy blng each ete hda vdi vinyl etyl ete trong dietyl ete xuc tac p-toluen sunfonic axit d 0-5 "C trong ddng khi tra. Din xult nitril sau khi da bao vp nhdm cacbonyl va hydroxy V, chiing tdi thyc hipn phan img metyl hda bang CH3Li, sau dd giai phdng cac nhdm bao vp bang dung dich axit HCl. Phan inig metyl hda xay ra rat manh liet, can rlt thgn trgng khi thyc hipn. San pham A'-hydroxyprogesteron VI nhan dugc vdi hipu suat 93 % (tinh tir chat IV), dugc giam dinh cau triic bang cac pbuong phap phd, dge bipt trdn phd ' H - N M R xuat hipn tin hipu cgng hudng Clia nhdm metyl-acetyl d 2,75 ppm (s, 3H, CH3-2I).
CN CN
^ ^fiiC
Sadd I: Chuyin hoa 9a-0H AD dSn 21-acetat pregna-4,9(ll)-dien-l7ci,21-diol-3,20-dion Trong so do tong hop cocticoid, giai doan tao
m^ch nhanh cocticoit tCr pregnan la mot trong nhu-ng giai do^n quan trgng da dugc chiing toi l^ra chgn th\rc hi?n di qua giai d o ^ tao dan xuat 21-mono iot, sau 66 21-axetyl hoa bSng tac nhan CH3COOK.
Hi?u suat cua ca giai dojin nay la 85 %. CSu true san phim 21-axetat pregna-4,9(ll)-dien-17a,21-diol- 3,20-dion da dugc IdiSng djnh d^ra tren eac dir li?u ph6. DJe bi?t tren ph6 'H-NMR xuSt hien tin hieu cgng huong nhom metylen a 4,89 va 5,05 ppm (2 d, 2H,CH2-21,y=16Hz).
Tit ca cac phan img chiing toi da th^rc hign deu d?t hieu suat tot, tinh che kha don gian, diSu nay rat CO ^ nghla thuc tiln.
DS t6ng hgp 21-acetat pregna-4,9(ll)-diene- 17(1,21-diol-3,20-dione tir 9a-0H AD voi hieu suat tot doi v6i timg giai do?n. Cau tnic cua chat diu, cac san phim trung gian va san pham cuoi da dugc xac djnh bang c4c phuong phap ph6: IR, MS, H- va
"C-NMR.
LOT cam OTi: Nghien ciru nay nhgn du^rc tai lr(? ciia BQ Khoa hqc va Cong ngh?: De tai doc igp cdp Nha nuoc (DTBL.2011-G/49) vd Quy phdt triin khoa hoc vd cong nghi quoc gia (NAFOSTED) (de tdi ma so 104.01-2012.20).
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Lien h^: Liru Dire Huy
Phong thi nghiem Hoa hgc Stergid va Alkaloid, Vign Hoa hgc, Vign Han lam Khoa hgc va Cong nghg Viet Nam S6 18, Hoang Qu6c Vigt, Ciu Giiy, Ha N$i Email: ldhuy{gich.vast.vn.
