CH EM. RES. CH INESE U.

2006, 22( 1), 51 55

An Improved SyntheticM ethod of Saquinavir

* *

YANG Q in gang, HE Xu chang, LEI L i jun, ZHOU Q i tin g and BA I Dong lu

Shanghai Institute of M ateria M edica, Shanghai Institu tes of B iolog ical Sciences,
Chinese A cademy of S ciences, Shanghai 201203, P. R. China
R ece iv ed M arch 1, 2005

An im proved syn thetic m ethod o f saqu inav ir, an H IV protease inhib itor, is descr ibed. In com parison w ith the
m e thods in the repo rted w orks, the im proved procedures had seve ra l adv antages, such as less expensive agen ts, shor
ter reac tion tim e, and a sm aller am ount of the solven t needed. T o m easure the optica l pu rities o f the products, the in
termed iates we re de term ined by m eans o f chira lH PLC. Som e of the interm ed iates can a lso be used fo r the preparation
of new pro tease inhib itors.
K eywo rds H IV; Protease inh ibito r; Saqu inav ir; Synthesis
A rticle ID 1005 9040( 2006) 01 051 05

In troduction
The hum an im munodeficiency virus( H IV ) is the
causat iv e agen t of acquired imm unodef iciency syndrom e
( A ID S). S in ce H IV pro tease is essen tial for the repli
ca tio n and m atu ra tio n o f H IV, the in act iv ation of H IV
pro tease by eith er m utation or chem ical inh ib it io n leads
to the product ion o f imm ature, noninfectious viral parti
[ 1]
cles . Saqu in av ir( com pound 1) , anH IV protease in
h ib ito r, w as f irst approved by the FDA in 1995. Sa
quinav ir inhibitsH IV protease w ith aK i va lu e of 0 12
[ 2]
nm o l/L and show s a specificity for the enzym e . Be

cause the structure o f saqu in av ir conta in s six chira l

centers w ith N containin g heterocyc les, large scale
preparat io n in an enant io pure form poses consid erable
challenges for chem ists in synth etic fie ld.
Severa l synth etic routes to saqu in av ir have been
[ 3]
published . Retro synthetic ana ly sis show ed that it
could be assemb led by fragm ents 2, 3 and 4, w ith the
corresponding L am ino acids 6 and 7 as the starting m a
terials( Schem e 1). T he stereose lective synthesis of the
hydroxy l eth y lam ine core ( 3) is very cruc ia .l Further
m ore, the retention of the configurat io ns o f the am in

Schem e 1 R etzosyn thyic analysis of saqu inav ir.

* Supported by the Science and T echno logy Comm ission o f Shangha iM un icipa lity( N o. 024319126) .
* * T o w hom co rrespondence shou ld be addressed. E m a i:l d lba@
i m ai.l shcnc. ac. cn

52

CHEM. R ES. CH INESE U.

oac id deriv atives are part icularly challeng in g in the

preparat ion of pept idom im etic inhibitors.
In the presen t paper, w e describ e an im proved
synthetic m ethod o f saquinav ir( Schem e 2 ), w hich is

V o.l 22

like ly m ore pract ica l in scale up preparat io n because o f

the reduced costs. Com pared w ith th e re po rted w orks
[ 3]
in litera tu re , several procedures have been im proved
by us.

Sch em e 2 Th e syn the tic rou te for saqu in avir.

R eagen ts and react ion cond it ion s: a. N hyd roxysuccin im id e, E t3 N, ( C 6 H 5 O ) 2 P( O ) C ,l r. t. , 20 h, 93% ; b. L A sn H 2 O, N aHCO3 ,
50

60 ! , 3 h, 93% ; c. phthalic anhydride, tolu ene, reflux, 94% ; d. triphosgen e, 70 80 ! , 10 h; e. H 2, 10% Pd /C, 40 50 ! ,

17 h; .f N aHSO3 , r. t. , 8 h; g. N aCN, r. t. , 2 h, 70% ; h. 25% HC ,l reflux, 18 h; .i C lCOO E t, 40% N aOH, r. t. , 2 h, 59% ; .j 0 75

m ol /L N aOH, r. t. , 8 h, 89% ; k. CH 3 OH, H 2 SO4 , ref lux, 4 h; .l N aBH 4 , 0 ! , 88% ; m. 4 n itrobenzenesu lfonyl ch loride, N m ethy lm or
pho line, r. t. , 9 h, 94% ; n. ( HCHO ) n , 37% HC ,l 95 100 ! , 6 h, 80% ; o. 25% NH 3

H 2 O, 93% ; p. triphosgene, 60

70 ! , 3 h; q.

t BuNH 2 , r. t. , 14 h, 82% ; r. 3039 8 kPa H 2, 5% R u /C, 100 ! , 16 h, 65% ; s. N a2 CO 3, 4 methyl 2 p entanon e, reflux, 15 h, 95% ; t.
N aHCO3 , H 2 O; u. N aOH, ethanol and H 2 O, reflux, 5 h, 89% ; v. 2, HOO B t, D CC, TH F - 10 ! , 2 h, r. t. , 16 h, 93% .
( A ) syn th es is of fragm en t 2; ( B) synthesis of fragm ent 3; ( C ) syn th es is of fragm en t 4; ( D ) assem b ly.

R esu lts and D iscussion

In the synthesis of fragm en t 2, quina ld ic acid d i
hydrate( com pound 5) is preferab le in com parison w ith
anhydrous acid to the preparation of succ in im ide ester
( compound 8). T he yie ld o f com pound 8 in creases by

5% com pared w ith that in re.f [ 4] . T he ester( com

pound 8 ) w as coupled w ith L asparag in e 6. T he
[ 5]
qu in arg ine 2 w as provided in a yie ld o f 93% .
T he preparation o f o ther tw o fragm ents( 3 and 4)
w ere both started w ith L pheny lalan in e 7. A suspension

N o. 1

YANG Q in gang et al.

of L phenyla lanine and phthalic anhydr ide in to lu ene

w as refluxed to rem ove th e produced w ater w ithout ar
gon protect io n. ( S ) 3 Pheny l 2 phtha lim id o propion ic
acid 9 w as ob tained as w hite crystals. Conversion of
acid 9 to its acy l chloride 10 w ith triphosgene or thionyl
ch lo ride rather than the expensive oxa ly l ch lo r id e, fo l
low ed by hydrogenation on 10% Pd /C produced alde
hyde 11, w hich w as in stant ly converted into O hydro
C sulfonato adduct 12 w ith sodium bisulfite. Crude ad
duct 12 w as trea ted w ith sod ium cyanide to form cyano
hydrin 13, as a 31 m ix ture o f diastereom ers in favor
of the desired d iastereom er. Conversion of 13 to the
corresponding carboxy lic acid m ixture ( 14) by ref lu
x ing in 25% hydrochloric acid, follow ed by the reac
t io n w ith ethy l ch lo ro form ate affo rded N protected acid
15, wh ich w as purified as the sin gle diastereom eric iso
m er by recrystallizat io n in toluene. Stereo isom eric pure
fragm ent 15 w as converted into oxazolidin one 16,
w hich w as dem onstrateded as the cis isom er by the cou
pling constant( 8 72 H z) of HC ( 4) HC ( 5) in the ox
1
[ 6]
azo lid inone rin g in th e H NMR spectrum . Interm e
diate 16 was then esterified to form the correspond ing
m ethyl ester ( 17), w hich w as reduced w ith NaBH 4 to
g iv e a lcohol 18.
In order to check the optical purity o f the prod
ucts, w e a lso prepared enant iom eric 18, ( 4R, 5R ) 4
benzy l 5 hydroxym ethy l oxazo lidin 2 one,
from the
corresponding D am in o acid. The enant iom eric excess
of( 4S, 5S ) 4 benzyl 5 hydroxym ethy l oxazo lid in 2 one
18 is nearly 100% determ ined by H PLC w ith a ch iral
stationary phase column. Because the tw o ch ira l cen
ters in key in term ediate 18 w ill be no t racem ized, the
reactio n products in the nex t steps could be purif ie d by
conventional m ethods.
Esterif ic ation o f fragm ent 18 gave p nitrophenyl
su lfonate 3 in a total y ield of 28% from L pheny lala
[ 7, 8]
n in e .
For the preparation of fragm ent 4, L pheny lala
n in e w as also used as the start ing m ateria.l P ictet
Spengler cyclization of L pheny lalan in e 7 and parafo
rm aldehyde proceeded sm oothly in 37% hydrochloric
acid to prov ide sa lt 19 in a y ield of 80% . W hen the
reactio n temperature w as raised from 50 ! to 95 100
! , the reactio n t im e w as sho rtened to 6 h. No racem i
zation o f the product w as observed. T reatm ent of sa lt
19 w ith 25% aqueous amm on ia gave acid 20 in a 93%
y ie ld. A cid 20 reacted w ith tr iphosgene affo rd ing 21,
w hich w as treated w ith t BuNH 2 to form am id e 22 in a
82% y ield. H ydrogenation o f 22 w as perform ed on a
5% Ru /C catalyst under pressure to afford decahydro

53
[ 9]

isoqu in oline am id e 4 , in wh ich tw o new ch ira l cen

ters w ere form ed in desired con fig urations, in a 58%
y ield after recrysta llizat io n in petroleum ether ( 60
[ 10, 11]
90 ! ) instead of the expensive n hexane
.
F in a lly, fragm ents 2, 3 and 4 w ere assem bled
w ith each other via tw o coup ling reactions. p N itrophe
ny l su lfonate 3 and decahydroam id e 4 w ere condensed
in 4 m ethyl 2 pentanone to produce crystallin e salt 23
in a good y ield, wh ich w as further hydrolyzed to
[ 7]
am ino alcoho l 24 . T he fina l coupling reaction o f
segm ents 2 and 24 w as effic iently achieved w ith DCC
in the presence o f hydroxy 3, 4 d ih ydro 4 oxo 1, 2, 3
benzotriazine( HOOB t). Saqu in av ir 1 w as produced in
93% yie ld w ith 98 8% purity( HPLC ) , w hose physi
cal and spectra l data are coinc id ent w ith the reported
[ 12]
ones .

Conclusion
In conclu sio n, w e have com pleted an im proved
syn th etic m ethod of saqu in av ir, wh ich is lik ely m ore
favourable in the sca le up preparation. In com parison
w ith the reported works, several steps have been im
proved. For exam ple, the expensive agents can be re
placed by cheaper ones; the reactio n t im e can be
m arked ly shortened; and a w ater separator w as adopted
to save th e solvent during azeotropic dehydration. T he
optica l purity o f the in term ed ia tes can be determ in ed
w ith ch ira l HPLC. Som e interm ediates can be also
used for the preparation of new protease in hib ito rs,
wh ich is underway in our labora tory.

Experim ental
1

T he H NMR spectra w ere reco rded on a Varian

M ercury 400 MH z FT NMR spectrom eter. The data are
reported as va lu es in parts per m illion relat iv e to
TM S. T he M S spectra w ere obta in ed on a F inn ig an
MAT 95 m ass spectrom eter( E I: 70 eV ). T he optica l
rotat io n va lu e [ ] D w as determ ined by P erk in E lm er
341 a t 589 nm. T he m e lting po ints( uncorrected) w ere
determ ined w ith a Buch i 510 apparatu s. T he so lv ent
w as rem oved by rotary evaporatio n under reduced pres
sure. Anhydrous so lven ts were obtained by d istillation
over sod ium w ire.
1
Preparation of Quinaldic Ac id Succin im idyl
Ester( Fragm en t 8)
Quina ld ic acid d ihydrate 5( 2 09 g, 0 010 m ol)
w as in troduced in to 30 mL o f acetone, to w hich
N hydroxysuccinim ide( 1 27 g, 0 011 m o l) w as add
ed. Under stirring a solution of triethy lam in e( 3 5 mL,
0 025 m o l) in 10 mL o f acetone w as then added to the
suspension w ith in 5 m in, and the m ix tu re w as stirred at
room tem perature fo r 30 m in. A so lu tion of d ipheny l

54

CHEM. R ES. CH INESE U.

ch lo rophosphate( 2 5 mL, 0 012 m o l) w ithin 10 mL of

acetone w as then added dropw ise in 10 m in, and the
reactio n so lution w as turned to red w ith slig htly exo
therm a l change and salts precipitating. The m ix ture
w as stirred at room tem perature for 20 h until the reac
t io n w as com plete. T he suspension w as concentrated
in vacuo, and the resid ue w as put into 50 mL of eth a
nol and the so lu t io n w as st irred at room tem peratu re for
1 h. T he so lid w as then filtered of,f and w ashed w ith
ethano.l E ster 8 w as obtain ed as a pink powder( 2 53
g, y ield 94% ) , m. p. 190 192 ! . { re.f [ 4] 193
196 ! }.
1
H NMR ( 400 MH z, CDC 3l ) , : 8 36 ( d, J =
8 5 H z, 1H ) , 8 31( d, J = 8 4 H z, 1H ) , 8 21( d,
J = 8 5 H z, 1H ) , 7 91 ( d, J = 8 2 H z, 1H ) ,
7 85 7 81( m, 1H ), 7 73 7 69( m, 1H ) , 2 95
( s, 4H, ) . E lem ental ana.l ( % ) ca lc. ( found for
C14H 10N 2 O4 ) : C 62 22 ( 62 22 ), H 3 73 ( 3 79 ) ,
N 10 37( 10 33) .
2 Preparation of( S ) 3 Phenyl 2 phtha lim ido pro
pion ic A cid( F ragm en t 9)
A suspension o f L pheny lalan in e 7( 82 6 g, 0 50
m o l) and phtha lic anhydrid e ( 74 1 g, 0 50 m o l) in
600 mL of toluene w as refluxed to rem ove the produced
w ater w ith a w ater separator fo r 8 h. T he resultant m ix
ture w as coo led to room tem perature and st irred for an
other 2 h. The solid separated w as filtered, w ashed
w ith to lu ene. A cid 9 w as obtained as a wh ite so lid ( 139
g, yie ld 94% ). m. p. 180 182 ! { re.f [ 13 ]
D
180 181 ! }. [ ] 20 = - 215# ( c = 1 05, m eth a
nol).
1
H NMR ( 400 MH z, CDC l3 ) , : 9 18 ( s, br,
1H ), 7 79 7 75( m, 2H ), 7 69 7 65( m, 2H ) ,
7 21 7 11( m, 5H ), 5 25 5 21( m, 1H ) , 3 61
( d, J = 8 8 H z, 2H ).
3 Preparation of( 2S, 3S ), ( 2R, 3S ) 3 ( 1, 3 D i
oxo 2, 3 d ihydro 1H isoindol 2 yl) 2 hydroxy 4
phenyl butyro n itrile( 13) w ith Triphosgen e
A suspensio n o f ac id 9 ( 29 5 g, 0 10 m o l) ,
triphosgene( 20 0 g, 0 067 m ol) and DMF ( 0 40 mL )
in 250 mL of to lu ene w as st irred at 70 80 ! for 10 h
until the reaction w as com pleted. T he so lv en t w as re
m oved in vacuo. C rude ( S ) 3 pheny l 2 phthalim ido
propiony l ch lo ride 10 w as obta in ed as a yellow so lid.
A cy l chlorid e 10 w as d isso lved in 250 m L o f to lu ene,
1, 2 epoxybutane ( 17 mL ) and 10% Pd /C ( 2 0 g )
w ere added to th e so lu tion. T he suspension w as hydro
genated for 16 h and then f iltered. The catalyst w as
w ashed w ith to lu ene. T he com b in ed f iltrate and w ash
ing w ere trea ted w ith a so lution o f sod ium b isulfite

V o.l 22

( 20 g ) in 100 mL of w ater under st irring at 25 ! . A f

ter 7 5 h, the organ ic layer w as separated and extrac
ted w ith w ater( 20 mL 3) . T he comb in ed aqueous
layersw ere w ashed w ith d ichlorom ethane( 20 mL 3).
D ich lo rom eth ane ( 150 mL ) and N aCN ( 10 g ) w ere
then added to the aqueous phase, and the m ix ture w as
stirred at 25 ! for 2 h. T he aqueous layer w as separa
ted and w ashed w ith d ich lorom ethane ( 20 mL 3 ).
T he comb in ed organic layersw ere dried over anhydrous
sodium su lfate, filtered, and the filtrate w as concen
trated to y ie ld nitrile 10( 21 4 g, y ield 70% ) as a off
wh ite pow der o f a 74 625 4 m ix ture o f the( 2S, 3S )
and( 2R, 3S ) isom ers( H PLC, W aters Nova Pak C18
co lum n, m obile phase: [ V ( H 2 O ) V ( M e th ano l) =
32] .
4 P reparation of( 2S, 3S ), ( 2R, 3S ) 3 ( 1, 3 D i
oxo 2, 3 d ihydro 1H isoindol 2 yl) 2 hydroxy 4
phenyl bu tyron itrile( 13) w ith Th ionyl Ch loride
A suspension of acid 9( 16 0 g, 0 054 m o l) and
thionyl chloride( 30 mL ) in 200 mL of d ichlorom ethane
w as refluxed at 60 70 ! fo r 12 h. T he so lv ent w as
rem oved in vacuo. T he residue w as d isso lved in 20 mL
o f d ich lorom ethane, to w hich 100 mL o f petro leum
eth er( 60 90 ! ) w as added. The m ixture w as con
centrated to a vo lum e of 20 mL under reduced pres
sure, and the separated so lid w as filtered, w ashed w ith
petro leum eth er( 20 m L 3) . A cy l chloride 10( 15 75
g) w as obtained as a w h ite so lid, w hich w as used to
the fo llow ing react io ns as describ ed above. Product 13
( 11 4 g, y ield 69% ) w as obtained as a m ix ture o f
( 2S, 3S ) and ( 2R, 3S ) isom ers.
5 Preparation of ( S ) 1, 2, 3, 4 Tetrahydro 3 iso
qu inoline Carboxylic A cid Hydroch lo ride( 19)
A suspensio n of L phenyla lanine 7 ( 12 0 g,
0 073 m o l ) and para fo rm a ld ehyde ( 4 65 g, 0 16
m ol) in 120 mL o f 37% concentrated hydrochloric ac id
w as st irred at 95 100 ! for 6 h. T he reaction m ix
ture w as coo led to 0 ! and the wh ite crystallin e so lid
w as filtered ou.t A fter be ing w ashed w ith co ld wa ter
and acetone, hydrochloride 19 ( 12 4 g, y ield 80% )
w as obtain ed as a wh ite powder.
1
H NMR( 400MH z, DM SO d6 ) , : 9 60( s, br,
1H ) , 7 28 7 24 ( m, 4H ) , 4 37 ( dd, J = 11 1,
5 1 H z, 1H ) , 4 27( s, 2H ) , 3 31( dd, J = 16 9,
5 1 H z, 1H ) , 3 13( dd, J = 16 9, 11 1 H z, 1H ) .
6 P reparation of Saqu inavir( 1)
A solution o f qu in arg ine 2( 14 37 g, 0 050 m ol)
and compound 24( 20 08 g, 0 050 mo l) in 150 mL o f
tetrahydrofuran w as cooled to - 10 ! , to w hich
HOOBt ( 8 15 g, 0 050 m o l) and DCC ( 11 0 g,

N o. 1

YANG Q in gang et al.

0 053 m ol) w ere added. The m ix ture w as stirred at

- 10 ! for 2 h and at room tem perature fo r 16 h, then
dilu ted w ith 150 m L of ethyl acetate. T he m ixture w as
filtered, and th e filtrate w as w ashed w ith a saturated
aqueous so lution o f sodium bicarbonate ( 50 mL 3) ,
saturated aqueous so lu t io n o f sod ium ch lo r id e( 50 mL
3) and dried over M gSO 4. The solvent of the o rgan ic
layer w as rem oved in vacuo, and the residue w as chro
m atographed on silica ge l by using 4% m ethano l in d i
chlorom ethane for the e lu t io n to g ive saqu in av ir
1(31 22 g, 93% ) as a yellow ish pow der in a purity of
98 8% ( H PLC, W aters Symm etry co lum n, [V (H 2O )
V ( M eOH ) = 15] .
+
M S ( ESI) , m /z: 671 [ M + 1 ] , 693 [ M +
+
D
23] . [ ] 20 = - 56 6#( c = 0 50, m ethano l). IR
- 1
( KB r) ,
/ cm : 3357, 2923, 2860, 1660, 1521,
-1
1
1498, 1454, 1226 cm . H NMR ( 600 MH z, CD3
OD ), : 8 46 8 43 ( m, 1H, ) , 8 15 8 13 ( m,
1H, ) , 8 12 8 10 ( m, 1H ) , 7 98 7 97 ( m,
1H ) , 7 82 7 80 ( m, 1H, ) , 7 68 7 65 ( m,
1H ), 7 18 ( d, J = 7 32 H z, 2H, ), 6 89 ( ,t J =
7 61 H z, 2H ) , 6 73 6 70 ( m, 1H ) , 4 92 4 90
( m, 1H ), 4 26 4 23 ( m, 1H ) , 3 89 3 86 ( m,
1H ), 3 03( dd, J = 11 7, 1 8 H z, 1H ), 3 00( dd,
1H, J = 14 1, 3 5 H z), 2 75( dd, J = 15 5, 6 4

55

H z, 1H ) , 2 70 2 60 ( m, 4H ) , 2 22 2 16 ( m,
2H ) , 2 06 2 00( m, 1H ) , 1 95 1 89( m, 1H ),
1 76 1 60( m, 3H ) , 1 54 1 46 ( m, 3H ), 1 28
( s, 9H ) , 1 16 1 37( m, 4H ) .
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