Note: Descriptions are shown in the official language in which they were submitted.
~-~ W092/05173 ~ O PCT/US91/06815
.. -- 1 --
J,
.. DESCRIPTION
'``'' .
' SUBSTITUTED TRICYCLIC COMPOUNDS
, ' ,
BACKGROU~ OF THE I~E~ION
Field of the Inventlon
The present lnventlon relates to certaln
substltuted tricyclic compounds whlch lnhlblt the
enzyme ;hymidylate synthase (~TS"), to pharmaceutlcal
compositions contalning these tricycllc compounds, ~nd
to the use of these compounds to lnhibit TS, lncludlng
all effects derived from the lnhibitlon of TS. Effects
; derived from the lnhibltion of TS include the
inhibition of the growth and prollferation of the cells
of higher organlsms and of mlcroorganlsms, such as
., yeast and fungl. Such effects lnclude antltumor
actlvity. A process for the preparation of the
substltuted trlcycllc compounds of the lnventlon ls
also dlsclosed.
The large class of antlprollferative agents
lncludes antlmetabollte compounds. A particular
subclass of antlmetabolltes known as antlfolates or
antlfols" are antagonlsts of the vltamln follc acld.
, 25 Typlcally, antlfolates closely resemble the structure
of follc acld, includlng the characteristlc p-benzoyl
glutamate molety of follc scid. TS has long been
considered an important target enzyme in the design and
., .
.
i .: ~. . .. : .
.. . ..
: ,
.. . .
W092/05173 PCT/US91/06815 ~
~ ~ rJ ~ V
synthesis of antltumor ~sents, and a number of folate
analogues have ~een sYntheslzad and studled for their
ability to lnhibit TS. See, for ~xample, Brl~ner et
al., Folate Analogues ~s Inhibitors of Th~mldvlate
SYnthase, J. Med. Chem. 30, 675 (1987): Joneq et al.,
Qulnazollne Antlfolates Inhlbitlnq Thvmidyl~te
SynthasP: ~en~oYl ~in~ ~.odl i~at~ons, J. Med. Chem.,
29, 468 (l986), Jones et al., ~uina~olln2 Ant ~olates
Inhlbltlng Thvmldvlats Synthas2: Ysriation Of tne
Amlno Acld, J. ~ed. C:~m., ~9, lIl~ ( Isa3 ~; ~ad Jones
et al., Qulnazollne AntlTolates ~nh$bitins Thvmldylate
SYnth2se~ Var~ation of the N~ Substituent, J. Med.
_
Chem. 28, 14~S (1985); ~n~ cope~dlng U.S. PatPnt
Appllcation Serlal No. 07/432,338 filed November 6,
1989.
SUMMARY OF THE INVENTION
The present lnventlon lntroduces a novel class of
~ubstltuted trlcyclic compounds whlch do not
partlcularly resemble the structure of olic acid and
yet, unexpectedly, lnhlblt the enzyme TS. The present
lnventlon also rPlates to pharmaceutlcal compositions
containing these substltuted tricyclic compounds and
the use of these compounds to lnhlblt TS, lncludlng all
effects derlved from the lnhl~ition of TS. Effects
derlved from the lnhlbitlon of TS lnclude the
lnhlbltlon of the growth and prollferation o the cells
of hlgher organlsms and of mlcroorganisms, 6uch as
yeast and fungi. Such effects lnclude antltumor
actlvlty. Processes for the preparatlon of the
6ubstltuted tricycllc compounds of the lnventlon are
~lso dlsclosed.
DETAILED DESCRIPTION OF THE INVENTION
The present lnventlon relates to antiproliferative
tricycllc compounds capable of lnhibiting thymldylate
synthase havlng the Formula (Q):
'
.
~ v ~
WO92/0~173 PCT/US91/06815
C C X
/ \ / \\ /
U C C
(CH,)" ¦ ¦ ( Q )
` 10 I / \ // \
V- ¦ -W CH Y
2 1 1
A
Ar - B
whereln:
X and Y form a five- or slx-membered heterocycllc
ring contalning at least one nitrogen;
Z is a hydrogen, halogen, car~on, oxygen or
nltrogen atom;
U ls a carbon or nitrogen atom;
n ls 0 or the lnteger 1;
V is a carbon or nitrogen atom;
W ls a carbon or nitrogen atom;
-. A ls in the l- or 2-posltion and is a nitrogen
. atom, a sulfur atom, or a substituted or
unsubstituted alkylene group;
Ar is an aryl or heteroaryl group having one or
more rings; and
ls either (1) an oxygen or nltrogen atom, or a
-C0- or -S02- group, any of whlch ls linked to
an amlno acld, aryl group, heterocyclic group
or alkyl gr~up, cr (11) a substituted or
. 35 unsubstltuted alkyl group.
As used hereln, the expression ~a compound capable
- of lnhibltlng thymldylate synthase~ denotes a c~mpound
~i wlth a TS lnhibltion constant X, of less than or equal
to about lO'M. The compounds of the lnventlon
preferably have K, values ln the range of less than
about lO-sM, preferably less than about lO-6M, even more
:~;
.
:
~.
. ,
W0~2tO5173 PCT/US91/06815 ._
u ~
- - 4 -
preferably les~ than about 109M and, 30st preferably,
ln the range from a~out 10~2 to a~out lOl~M.
X and Y in ~ormula (Q) can for~ any ~lve- or ~x-
: membered heterccyclic rlny oontalnln~ at 1 3a3t one
nitrogen such as, for example, pyrrole, lmidazole,
pyrazole, pyrldlne, pyrazin2, pyrl~ldin2, and
pyrldazlne rlngs. Preferzbly, X and Y form the rlng:
N
\ / \\
C \~
C ~ -~
C
/ \ /
wherein P ls hydrogen; a lower alkyl g-oup such as
methyl, ethyl, propyl, lsopropyl, tert-butyl and the
like; or an amlno group -NRlR2 wherein R and R2
lndependently represent hydrogen, alkyl, amino,
hydroxyl, or the like.
Z $n Formula (Q) can ~e a hydrogen atom; a halogen
atom such as chloro, bromo, or fluoro; a carbon atom
which, taken wlth other appropriate atoms, may form
such groups as subst~tuted or unsubstituted alkyl,
alkenyl, alkynyl, alkyl-oxy-alkylene, allyl, benzyl,
acetyl, carbamyl, carbalkoxy, cyano, phenylacetyl,
~mlnoalkyl or the llke: an oxygen atom whlch, taken
together wlth other approprlate atoms, may form such
groups as hydroxy, alkoxy, oxamldo, oxamyl, acetoxy,
phenoxy, phenylsulfamyl, phenylsulfonamldo or the llke;
or a nltrogen atom which, taken with other approprlate
atoms, may form such group6 as amlno, nltro, acetamldo,
anillno, benzamido, formamldo, hydrazlno, hydroxamlno,
lsocyano, n~tramino, nltroso, oxamido, sulfamldo,
alkylamlno, or the llke. Preferably, z 18 a hydrogen
atom.
The lnteger n in Formula (O) can be O or 1, but ls
preferably 0. In other words, the left-hand rlng ln
,
W092/05173 PCT/US91/06815
, .
- 5 -
the formula as wrltten above 1~ elther a 6- or 7-
mem~éred ring.
U, V and W ln Formul~ ~Q) are each lndependently
carbon or nltrogen atoms and, taken together with ( C~2 )n
and other spproprlate carbon atoms a6 lndlcated ln the
formula, form a 6- or 7-membered ring such as, for
example, a benzene, cyclohexene, pyrldlne,
tetrahydropyrldlne, pyridazlne, pyrlmidlne, 1,2,3-
trlazine, cycloheptene, tetrahydroazeplne or thP liXz
rlng.
In a preferred embodlment, U ls a carbcn atom,
ls a carbon atom, and W ls a nltrosen atom. Most
preferably, U, V and W are, taken together with (CH2)~
and one or more other carbon ~toms as lndlcated ln the
above formula, form a rlng havlng the following
structure:
CH2 . ,~
\ /
: CH2 C
l l
CH~ C
/ \
N
(A - Ar - B)
Generally, A ln Formula (Q) above ls in the 1- or
2-posltion of the rlng formed by U, -(CH2)~, V, W and
the other appropriate atoms referred to above. A can
be a nltrogen atom which, taken wlth other approprlate
atoms, may form such groups as dl- or trlsubstltuted
emlne or the llke; a sulfur atom whlch, taken wlth
other approprlate atoms, may form such groups as a thlo
llnkage (^S-), thloalkylene, thloamide and the llke: or
any substltuted or unsubstltuted alkylene group such
a5 ~ for example, methylene, ethylene, n-propylene,
lsopropylene, n-butylene, tert-butylene, n-hexylene or
the llke. It should be noted that, lf A ls sulfur,
then W and V should be carbon to produce a reasonably
stable compound. Preferably, A is a substituted or
W092/05173 ~ v~ ' 6~ PCT/US91/06815 -
.
- 6 -
unsubstltuted alkylene group such a8, for zxample,
: methylene, ethylene, n-propylane, lsopropylene, n-
butylene, tert-butylene, n-hexylene, or the like.
As lndicatad above, .~r can ~e ~n~ one o~ ~ large
num~er of aryl or heteroaryl groups having one or more
rlngs. Examplzs of u32 'ul aryl sing s-oups inc'ude
phenyl, 1,2,3,4-tetrahydronaphthyl, naphthyl,
phenanthryl, anthryl ~nd the l~Xo. _xzTI2l~s o~ t~ical
heteroaryl groups lnclude ~-~=m~2-ed ~,cr.oc,c'ic _inS
groups such as thienyl, pyr~oiyl, ~n-~y~r.olyl,
lmidazolyl, pyrazolyl, su~yl, iso~ iazolyl, fura~anyl,
isoxazolyl and the llke; 6-mamD2rod ~onocyclic groups
such as pyridyl, pyranyl, pyrazinyl, pyrlmidinyl,
pyridazinyl and the llke; and polycyclic heteroaryl
groups such as benzo[b]thienyl, naphtho~2,3-b]thlenyl,
thlanthrenyl, lsobenzofuranyl, chromenyl, xanthenyl,
phenoxathiinyl, lndollzlnyl, lsolndolyl, 3H-lndolyl,
lndolyl, lndazolyl, purlnyl, 4H-quinolizlnyl,
isoqulnolyl, qulnolyl, phthalazl~yl, naphthyrldlnyl,
qulnoxallnyl, qulnazollnyl, cinnollnyl, pterldinyl, 4H-
carbazolyl, carbazolyl, beta-carbollnyl,
phenanthridinyl, acrldinyl, perimidinyl,
phenathrollnyl, phenazinyl, lsothiazlyl,
phenothiazinyl, phenoxazinyl and the llke. Preferably,
Ar ls a monocycllc or blcycllc aryl group, such as
phenyl or naphthyl.
~ , a substituent on the Ar group discussed above,
can be an oxygen atom whlch may be taken alone to form
an ether llnkage (-0-) or whlch ~ay be taken together
wlth other approprlate atoms to form such groups BS
hydroxy, alkylenoxy, oxamldo, oxamyl, ~cetoxy, phenoxy,
phenylsulfamyl, phenylsulfon~mldo or the llke; or a
nltrogen atom which, taken wlth other approprlate
~toms, may form ~uch groups as ~mino NR'R2 whereln R
and R2 can each lndependently ~e alXyl, or the like,
nltro, acetamldo, ~nlllno, benzamldo, formamido,
hydrazlno, hydroxamlno, lsocyano, nitramlno, nltroso,
- W092/0~173 , PCT/US91/06815
~v153~
oxamido, sulfamldo or the llke; or e -C0- or -SO2-
group.
Any one of the dlv~lent B groups above m~y be
further llnked to an amlno acld ~roup such as alanlne,
arglnine, asparglne, asp~rtlc acld, cystelne,
slut~mlne, glutamlc acid, ~lyclne, hlstldlne,
lsoleuclne, leuclne, lyslne, methlonlne, phenylal~nlne,
proline, serlne, threonine, tryptophan, tyroslne,
V811n2 or the llke; an aryl group such as phenyl,
naphthyl and the llke; a heterccycllo sroup such as
thienyl, pyrrolyl, imidazelyl, pyrazolyl, lsothiazolyl,
pyrlmidinyl, pyrazinyl, tetrahydroyrazinyl,
benzo[b~thlenyl, naphtho~2,3-b~thlenyl, phoxathilnyl,
. ~ndazolyl, phthalazlnyl, clnnollnyl, carbollnyl,
phenanthrollnyl, phenoxazlnyl, and the llke; or an
alkyl group such as methyl, ethyl, n-propyl, lsopropyl,
n-butyl, tert-butyl, n-hexyl and the like. Flnally, B
lt~elf may be an alkyl group such as methyl, ethyl, n-
propyl, lsopropyl, n-butyl, tert-butyl, n-hexyl or the
llke.
In a preferred embodlment, ~ ls an -C0- or -SO2-
group llnked to an aryl or heterocycllc group,
preferably phenyl or tetrahydroyrazlnyl. When
lncludes an aryl group, the aryl group may be
unsubstltuted or may be substltuted wlth one or more of
a wide varlety of electron-donatln~ and electron-
wltharawlng substltuents. ~yplcal substltuents lnclude
halogen, hydroxy, alkoxy, alkyl, hydroxyalkyl,
fluoroalkyl, amlno, -CN, -N02, carbalkoxy, carbamyl,
carbonyl, carboxyldloxy, c~rboxy, amlno acld carbonyl,
~mlno acld sulfonyl, sulfamyl, sulfanllyl, sulfhydryl,
ulflno, sulflnyl, sulfo, sulfonamldo, sulfonyl or the
llke. Most preferably, ~ 18 an -SO2- group llnXed to
phenyl group whlch 18 elther unsubstltuted or
substltuted ln the par~-positlon wlth ~ hydroxy group
or an alkoxy group such as methoxy, ethoxy, 150-
propoxy, tert-butoxy, chloroethoxy or the llke.
W O 92/05173 4~ 9 ~ PC~r/US91/06815 --
P~rtlcularly preferred structures for -Ar-8
lnclude: .
CH - CH CH2-CH2
. //, \\ / ~
~ C C ~ SO2 ~ N ~rH
CH ~ CH C ~-CH2
CH - CH CH - 5~
// \\ // \\
~~ C C ~ SOz ~ C C~ ;
C~ - C~ C-~ ~ C-~
CH - CH Cn - CX
// \\ // ~\
-- C C - SO~ - C C-OH
CH - CH CH - CH
CH ~ CH - CH ~ CH
// \\ // \\
~~ C C ~ SO2 ~ C C-OCH3
/
CH - CH CH ~ CH
CH CH
\ / \\ / \\
C C CH
~ I l
CH C C--CH2-OH
// \ //
CH CH
4a
CH ~ CH COOH
// \\ I
-- C C - CO-NH-CH-CH2-CH2-COOH
CH ~ CH
- ; ' . . : ' :
.
.
. ,. ,~
.: \
WO92/~5173 ~ ~J l ~ 9 0 PCT/US91/06815
CH CH
\ // \ / \\
C C C - O-~ whereln R 18 H or
¦ alkyl, such a8
S --- C CH -methyl; and
\ ~/
C~
CH CH2-OH
\ / \\ /
'~ . C C
CH C
i \ // \
C T wherein ~ is H or -CN.
, 1.
CH2-OH
In addition to the substltuent B defined above, Ar
in Formula (Q) can also be substltuted with one or more
of a wide variety of electron-donating and electron-
withdrawing substituents. Typical substituents include
halogen, hydroxy, alkoxy, alkyl, hydroxyalkyl,
fluoroalkyl, amino, -CN, -NO2, carbalkoxy, carbamyl,
carbonyl, carboxyldioxy, carboxy, amlno acld carbonyl,
i~ amino acid sulfonyl, sulfamyl, sulfanilyl, sulfhydryl,
`~ sulfino, sulfinyl, sulfo, sulfonamido, sulfonyl or the
~l 30 like. Preferably, these addltional substituents are
`, selected from the group consisting of -CN, fluoroalkyl,
- and sulfonyl.
~` In a partlcularly preferred group of compounds
capable of lnhibitlng thymidylate synthase accord$ng to
the present lnventlon, X and Y in Formula (Q) form the
rlng:
N
\ / \\
i.' C\\
I C--P
Ct
.. t \
; NH
; 45 wherein P is lower alkyl such as methyl, amlno or
hydrogen; U is carbon; n is 0; V is carbon; W is
.
.~ ' ' . '' ~ : , .
.
~, :. . . . . ...
.: , ` ~ . .. ~' ` . .
.. ~ :. ................... ~ , ,
.
W092/05173 ~ O PCT/US91/06815 --
- 10 -
nltrogen; A 16 ~ethylene; and -Ar-B i8 selected from
the group consistlng of:
CH - CH CH2-C'A~
t/ \\
- C C - SO2 - ~ ~H
CH ~ CH CH2-Cnz
CH - CH Cr: - C.Y
// \\ /,/ ~\
-- C C ~ SO2 ~ C C~ ;
\, / ~ /
CH - CH CH - CY
CH - CH Cr - CH
// \\ // \\
-- C C - 52 - C C-OH
CH - CH CH ~ CH
CH - CH CH - CH
// \\ // \\
-:~ -- C C - SO2 - C C-OCH3
-. 30
CH - CH CH - CH
CH CH
~ 35 \ / \\ / \\
C C CH
" ~ I I~ CH C ~C--CH2-OH
-'~ \ // \ //
. 40 CH CH
CH - CH COOH
// \\
-- C C - CO-NH-CH-CH2-CH2-COOH
CH - CH
.
., , , ,~, .
: ,. .~ ~ , . . .
,
,
:- . ,
~~~ W092/05173 i.vJ i ~J~ PCT/US91/06815
CH CH
\ // \ / ~\ '
C C C - O-R whsreln R 18 H or
l l ¦ alkyl, ~uch a8
S --- C CH oethyl: ~nd
\ //
CH
j CH CH2-OH
\ / \\/
C C
lS CH C
\ // \
-C T wherein T ls H or -CN.
I
CH2-OH
Examples of partlcularly preferred compounds of
the lnventlon include those falllng withln the a~ove
~roup whereln P ls hydrogen and -~r-B ls
CH - CH CH2-CH2
~ // \\ / \
~ -- C C - SO2 - N NH
.~ \ / \ /
CH - CH CH2-CH2
' 30
whereln P ls -NH2 and -Ar-B ls
,~
~ CH - CH CH2-CH2
// \\ /
:~ 35 - CC - SO2 - N NH
\ / \ /
. CH ~ CH CH2-CH2
,
wherein P ls -NH~ and -Ar-B 18
;, CH - CH CH - CH
: // \\ // \\
-- C C - SO2 - CCH
`t CH ~ CH CH ~ CH
;~
,..:
. ~ ~. .. . .. ..
: . . -
::, . . . .
. . .
- : ~
WO92/05173 ~ ~ J i ~ ~L) PCT/US91/~6815 --
- - 12 -
whereln P is -NH2 and -Ar-B 18
CH CH
\ / \\ / \\
5C C CH
CH C C--CH2-OH
\ // \ //
CH CH
whereln P ls -NH2 and -Ar-B ls
CH - CH C'n - CH
// \\ // \\
-- C C - SO2 - C C-Or: ;
\' / \ /
CH ~ CH CH - CH
whereln P ls - CH3 and - Ar - B i s
CH - CH CH - CH
// \\ // \\
-- CC - SO2 - C C-OCH3
; 25
CH - CH CH - CH
wherein P is -NH2 and -Ar-B is
. CH - CH CH - CH
// \\ // \\
:' -- C C - S2 - C C-OC~13
/
. 35 CH - CH CH ~ CH
:.
whereln P ls -NHCH3 and -Ar-B ls
CH - CH CH - CH
: // \\ // \\
.~. -- CC - SO2 - C C-OCH3
/
CH - CH CH - CH
:
;; whereln P ls -NH2 and -Ar-B ~s
., .
CH ~ CH COOH
// \\
.. -- C C - CO-NH-CH-CH2-CH2-COOH
CH ~ CH
s 55
.
.
.
.
:` ' ' . ' . , . . ~ . '.:
.
- WO92/05173 PCT/US9t/06815
- - 13 -
wherein P ls -NH2 snd -Ar-B 18
CH CH
\ // \ / \\
C C C - O-~ whereln R 18 H or
¦ ¦ 81kyl, such ~B
S --- C CH methyl; snd
\ //
CH
wherein P is -NH2 and -Ar-B i8
CH CH2-OH
\ \ /
C C
CH C
\ //\
C T wherein T is H or -Ch'.
: 20
' ' CH2-OH
. In another particularly preferred embodiment
sccordlng to the present inventlon, X snd Y ln Formula
.~ 25 (Q) form the rlng:
: CH
~ 1 3
:~ N
\ / \
i 30 C
~ 11 C - - P
:i C //
; / \ //
N
.` 35
:~ wherein P ls selected from the yroup conslsting of
-NH2 and methyl; U ls carbon; n ls O; V is carbon: W ls
nltrogen: A is methylene: ~nd -~r-~ is
CH - CH CH - CH
// \\ // \\
; -- C C - SO2 - CC-OCH3
J
CH ~ CH CH ~ CH
: 45 In a stlll further preferred group of compound~
sccording to the present invention, X ~nd Y ln Formula
(Q) form the rlng:
:
WO92/05173 ~ u~ i ~ 9 ~ PCT/US91/06815 -
- 14 -
\ / \\
I C--P
C
/ \ /
NH
whereln P l~ selected from the group conslstlng of
-NHOH and -NHNH2; U 18 carbon; n ls 0; V ls carbon; '~ is
nltrogen; A ls methylene; and -Ar-B ls
C~ - C~ C~ - C~
/~ \~ // \\
-- C- C - SO2 - C C-OC~3
CH ~ CH CH - CH
;In 8 stlll further partlcularly preferred compound
:~ 20 capable of lnh~bitlng thymldylate synthase according to
;the present lnventlon, X and Y ln Formula (Q) form the
rlng:
. N
: \ / \\
C \\
- C--NH2
,. C
/ \ /
NH
U ls carbon; n ls 0; V ls carbon; W ls carbon; A ls
dlvalent 6ulfur; and -Ar-~ ls
CH - CH CH - CH
':,' // \\ ~ \\
-- C C - SO2 - C CH
\ /. \ /
CH ~ CH CH ~ CH
The lnventlon also relates to a process for making
: 40 the compounds of the present lnventlon comprislng the
8teps of:
(l) allowlng a compound of the formula ~-Ar-A-D,
whereln A, Ar and ~ are a6 deflned above and
D 18 a dlsplaceable group, to react wlth a
compound of Formula (I):
. '
'
;
:.
W092/05173 ~ ~ J 16 J i~ PCT/US91/06815
- 15 -
C C X-precursor
\ / \ \ /
~ C C
(CH,)n I C (I)
/-\ // \
Y~ CH Y-precursor,
wherein X-precursor and Y-precursor are groups whlch,
when cycIl~ed with each other, form a five- or 8iX-
membered heterocyrlic ring containing at least one
nltrogen, to form a ~ubstltuted compound having the
fo~ul~:
z
C C X-precursor
U C C
1 / \ // \
V-¦-W CH Y-precursor; and
~ 2
.~ A
Ar - B
(2) cyclizing X-precursor and Y-precursor to form
with each other a flve- or ~lx-mem~ered
heterocyclic rlng contalnlng at least one
nltrogen atom.
The dlsplaceable group D can be any group whlch ls
displ~ceable under the re~ctlon condltlons used end ls
typlcally a halogeno ~uch ~s fluoro, chloro or bromo:
8ubstltuted sulfonyloxy such as ~ethanesulfonyloxy,
trlfluoromethanesulfonyloxy, toluene-p-sulfonyloxy, or
4-bromobenzenesulfonyloxy; an ~ldehyde: or the llke. A
halogeno 18 a preferred dl~placeable group, wlth bromo
belng partlcularly preferred.
The flr~t step, reactlng B-Br-A-D wlth a compound
of Formula (I), may be carrled out ln an organlc
.
WO 92/05173 ~ PCT/US91/06815
solvent. Typlcally, when an organlc ~olvent 18 used,
lt 18 an ~protic solvent such a8 dlmethylfor~mlde,
dlmethylacetamlde, dlmethylsulfoxlde, or
t~trahydrofuran. Especlally prefsrred solv~nt3 lnclud2
dlmethylformamlde and dlmethylacet~mide.
Typlcally, al80, the flrst step 18 c~rrled out ln
the presence of a weak base whlch wlll not lt~elf r~ac~
wlth one of the reactants. U3eful bases lnclude, .or
example, organlc bases such a8 a substituted ~mine sucA
as diisopropyl2thylamine, dlmethyl-sec-butyl~min~
methyl-~-Pthylaniline, N,N-dimethylanlllne or the l~Xe;
and lnorgan'c weak bsses ~uch ~s scdium, potassium
and/or calclum carbonate or the like. The reaction
temperature for the first step can vary from about room
temperature to about lOO-C, but preferably ranges from
about 65-C to about 85-C.
The second step of cycllzlng X-precursor and Y-
precursor wlth each other to form a flve- or slx-
membered heterocycllc rlng contalnlng at least one
20. nltrogen atom ls carried out ln the presence of a
cycllzlng agent such as HC(OCH3)~/HCl, CNBr,
CH3C(OCH3/HCl or a comblnatlon of a metal such as tin
and acetic acld. Most preferably, the cyclizlng agent
ls HC(OCH3)3/HCl or CNBr.
The reactlon may be carrled out ln the presence of
an organlc solvent, such as methanol, ethanol, butanol,
acetonltrlle, mlxtures thereof or the llke. When the
cycllzlng agent ls CNBr, for example, a m~xture of
methanol and acetonltrlle can advantageously be used.
In contrast, however, when the cycllzlng agent ls
HC(OCH3)3/HCl, t~e startlng materlal ls typlcally
dlssolved ln the HC(OCH3)3 ltself wlthout any addltlonal
solvent, and a relatlvely small amount of HCl ls then
added to lnltlate the cycllzlng step.
The temperature used for the cycllzing step ranges
from Just below room temper~ture to about 70~C, but is
preferably from about 20 to about 60-C.
.
' - . . ' ' " .'' ~'`' ~ ~
--; W092/OSl73 ~ v~ o PCT/US91/0681S
- 17 -
It ~hould be noted that one or more of X-
precur~or, Y-precursor, U, V and W may contaln a
chemical group or group~ whlch, elther before, after or
during the cou~se of elther the sub~tltutlon ~tep (1)
or the cyclizlng step (2):
(a) ~ay be protected by a protecting group or
(b) ~ay have one or more of any protectlng groups
present ramoved.
~ suitable protectlng group for a rlng nltrogen,
such as U, V cr ~ may ~e, for example, a
Di valoylox~l~ethyl group, which ~ay be removed by
hydrolysls with A base such as sodium hydroxide; a
tert-butyloxycarbonyl group, whlch may be removed by
hydrolys~s with an acid such as hydrochloric acid or
; 15 with a base such as llthlum hydroxide: or a 2-
(trlmethylsllyl)ethoxymethyl group, which may be
removed by a fluoride salt ~uch as tetra-n-butyl
ammonlum flourlde or with an acld such as hydrochloric
acid.
A sultable protecting yroup for a hydroxyl group
is, for example, an esterifying group ~uch as an acetyl
or benzoyl group, which may be removed by hydrolysls
wlth a base such a~ sodium hydroxlde. Alternatively,
when other groups pre6ent in the starting material do
not contaln an alkenyl or alkynyl group, the protecting
group may be, for example, an alpha-arylalkyl group
such as a benzyl group, which ~ay be removed by
hydrogenatlon ln the presence of a catalyst such as
palladium-on-c~arcoal or Ranay nickel.
A ~ultable protective group for a mercapto group
16, for example, an esterifylng group such a8 an acetyl
group, whlch may be removed by hydrolysls wlth a base
such as sodium hydroxide.
~ ~uitable protective group for an amino group may
be, for example, an alkylcarbonyl group such as an
acetyl sroup (CH3CO-) or a benzoyl group, which may be
removed by treatment with an inorganic acid such as
W092/0~173 ~ ~ PCT/US91/06815 i-^
- - 18 -
nltrlc, sulfurlc or hydrochlorlc acld, or by bsse
hydrolysis wlth sodlum hydroxlde. Another protectlve
group for an amlno group ls an alkoxycarbonyl group
3uch a5 a ~ethoxycarbonyl or a tsrt-butylo~ycarbonyl
group. These groups may be removed by treatment wlth
an organlc acld such a~ trlfluoroa~etlc acld.
Alternatlvely, the protectlve group ~ay be a
benzyloxycarbonyl group, whlch Day be removed by
treatment wlth a Lewls acld such as boron
trls(t-lfluo.oac~tats) or hydrogen gas ln the presence
of A palladium-on-charcoal catalyst.
~ suita~l~ protective group for a primary smino
group is, for example, an alkylcarbonyl sroup such as
acetyl, whlc~ may be removed by treatment wlth an
lnorganlc acld such as nltrlc, sulfurlc or hydrochlorlc
acld, or a phthaloyl group, whlch may be removed by
treatment wlth an alkylamlne such as 3-
dlmethylamlnopropyl amine, wlth hydrazine, or wlth
ammonla.
A sultable protectlve group for a carboxy group
may be an esterlfylng group, for example, a methyl or
an ethyl group, which may be removed by hydrolysls wlth
a base such as sodlum hydroxlde. Another useful
protectlng group 15 a tert-butyl group, whlch may be
rsmoved by treatment wlth an organic acld such as
trlfluoroacetlc acld.
Preferred protectlve groups lnclude an esterlfylng
group, an alpha-arylalkyl group, an alkylcarbonyl
group, a sub~tltuted or unsubstltuted alkoxycarbonyl
group, a phthaloyl group, a plvaloyloxymethyl group or
a methyl oxyether-type group such as methoxymethyl or
2-(trlmethylsllyl)ethoxymethyl.
A partlcular aspect of the lnventlon relates to a
proces~ of maklng a substltuted trlcycllc c~mpound
capable of lnhlbitlng thymldylate synthase from a
startlng compound of Formuln (I), whereln the compound
of Formula ~S) has the structure of Formula (II):
-- W092/0~173 PCT/US91/06815
~ u v i ~ O
-- 19 --
Z
C C NH-Ac
\ / \\ /
5 U C C
(CH2)n 1 1 (II)
:~ / \ // \
~ OV---W Cl~ ~2
wh~re~n ~c ls a CH3CO- protectlve sroup. A process of
ma~ing thls group of ~tartlng compounds may comprise
the s~eps of:
: 15(1) ~electively protectlng an amine compound of
~e~.~l2 III-
, Z
C C N}~2
/ \ / \\ /
U C C
(CH2)n
.C CH
, 25 ~ \ //
V---W CH
. to form the correspondlng acetamide; and
(2) nitrating the correspondlng acetamlde to form
; 30the compound of Formula (II).
::~In the flrst step, addlng a protective group to a
-compound of Formula II to form the corresponding
acetamlde 16 preferably performed by treatlng the amlne
compound of Formula ~III) wlth an approprlate anhydrlde
compound, such as acetlc anhydrlde, ln the presence of
an organlc solvent, cuch as pyrldlne or the ~e. The
addltlon of a protectlve group can occur at
temperatures lower or hlgher than room temperature.
~yplcally, however, the addltlon of a protectlve group
occurs at a temperature between about -lO and ~15-C
and, most preferably, between about -lO and -5-C.
~ he second step of nltratlng the acetamlde formed
by selectlvely protectlng the amlne compound can be
carrled out ln the presence of one or more of the many
,. ~
-, ~ : . :
W092/05173 PcT/uS91/o68ls -~
~-i '`'ib'9V
- 20 -
known nltratlng ~gents, such as (1) a mlxture of nltric
and sulfurlc aclds; (2) a ml~ture of nltrlc, ulfurlc
Hnd ~cetlc aclds; or (3) a mlxture of n~trlc acld and
ac~tlc anhydrlde. Preferably, the nltratlng agent 18 a
ml~ture of nltrlc and sulfurlc aclds. The nltratlon
~tep may be carrled out over a wlde range of
tsmper~tures but is typlcally carried out at e
tamperatur~ ~etween about -10 and ~lO-C, preferably
between about -10 and 5-C.
The ~mine com~ound of ~ormula (III) ltself can be
preparad by several dlfferent reactlon schemes. In one
~mbcdim~nt, the amine c~m~ound of Formula (III) ls
prepared by raducing a compound corresponding to the
amlne having one or more sltes of unsaturatlon ln the
6- or 7-membered rlng formed by U, V and W taken ln
comblnatlon wlth an approprlate number of carbon and
hydrogen atoms. In thls ~mbodlment, the reductlon can
be performed under wldely varylng reductlon condltlons,
but ls preferably carrled out ln water or ln an organic
solvent such as methanol, ethanol, tetrahydrofuran,
acetlc acid or the llke, ln the presence of a reduclng
agent such as a hydrazlne compound or hydrogen gas at a
pressure of at least one atmosphere, preferably at a
pressure from about l to about 50 psl. A reductlon
cataly6t 18 also used, such as platlnum oxide (as
described ln I~hlkawa et al., Chem. Pharm. Bull., 37,
2103 (1989) whlch 18 hereby lncorporated by reference).
In a mo6t preferred embodlment, the compound
correspondlng to the amine ha~lng one or more sltes of
unsaturQtion has the formula:
Z
CH C NH2
// \ / \\ /
CH C C
CH C CH
\\ / \ //
N C~
.
.
~ WO92/051~3 h V J ' ~ ~ ~ PCT/US~1/06815
- 21 -
. and 18 raducad by tre~t~ent wlth hydrogen 9~B and a
pl~tlnum oxlde cat~lyst ln an organlc ~olvent uch ~s
gl~clal ac~tlc acid.
Alt2rnatl~ely, ln another preferrsd embodlment,
the ~mlne compound o~ Formula (III) 1~ prepared ~y:
(1) nltrating an unsub~tltut2d compound of
: Formula (IY):
z
C ~-
: / \ / \~
U C C~
(~s2)~
' C C~
/ \ //
V---W CH
: 20
to form a nltrated compound having the formula:
~; Z
.,, C C NO2
- . / \ / \\ /
U C C
(CY~ )n ~ !nd
C CH
.' / \ //
V~~~W CH
(2) reduclng the nltrated compound to form the
amlne of Formula (III).
The reactlon condltlons for the nltratlon step (1)
are ~yp~ ;c~ . a~ v~ IO~ ;ral n~.tr~
reactlons. Preferably, however, the nltrat$on step ls
performed ln the presence of nltrlc acld and at a
temperature between about -10 to about 20-C. A
partlcul~rly preferred nltratlon process uslng these
condltlons ls descrl~ed ln Amlt et al., J. Chem. Soc.
Perklns II, 57 ~1976), whlch ls hereby lncorporated by
reference.
.~.
W092/05173 PCT/US91/06815 ~
- i 9
- 22 -
The reactlon condltlons used for the reductlon
step (2) can vary graatly but, typlcally, lnclude one
or morz cf the following: (a) treatment wlth SnCl~ in
the pr33_ncP of hydrochlorlc ~cld: (b) tra~t~ent wlth
zlnc ln the presence of ocetlc acld: (c) treat~ent wlth
Fe~(C0)~2 ~ n ~enzene ~nd ~eth2nol: (d) treatment wlth
~ydrogen gas ln the presenc2 of a palladlum-on-charcoal
catalys~; (o) tr~a.~Pnt ~ith hydrogen gas ln the
pr23encP oS a pl2tinum o~ld~ catalyst ln an organic
0 501vor.~ S"C~. ~5 gl3cl~1 acstlc a-~d, and ( f) treatment
~ith a hyàrazlno in the ~r~senc2 of ~ reduction
catalys~. Pre~_r~ely, ho~ever, the roduction 81ep ( 2 )
is carrisd OUt with hydrogen gas as the reduc~ng agent
ln the presence of a palladium-on-charcoal catalyst ln
an organic solvent, for example, An alcohol ~uch ~s
methanol.
Most preferably, the compound of Formula (IV) has
the formula:
.~ . z
~ 20
CH2 C
\ / \\
CH2 C CH
~ IV)
CH2 C CH
/ \ //
N CH
Ac
whereln Ac i8 an acetyl ( C~3CO- ) protectlve group which
18 removed between the nltratlon 6tep and the reduction
step. ~he semoval of thls protectlve group can be
accompllshed by the general metho~s descrlbed above for
removal of protectlve groups from mlno groups.
Preferably, the acetyl group 18 removed by treatment
wlth an lnorganlc acld such as hydrochlorlc acld ln a
solvent, for example, water, an alcohol such as
ethanol, or a mlxture of water and ~n alcohol at a
temperature about the boilln~ polnt of the solvent.
;~
: .: ....
... ... .
--~. W092/05173 ~ 0 PCT/US91/06815
As stated ~bove, X-precursor and Y-precursor are
groups whlch, when cycllzed wlth each other, form a
flve- or six-mem~ered heterocycllc rlng cont~lning ~t
least one nltrcge~. In a preforrad embodiment, in the
compound
z
C C X-prscurQor
\ / \\ /
10 U C C
2~
C C
, ~ " ~
15 y~ C.i Y-p_~cursor;
A
Ar - B
: 20
X-precursor is -NH-Ac and Y-precursor is -NO2. In this
embodlment, the -NH-Ac and -~2 groups can be cyclized
wlth each other uslng one of several alternatlve
:. methods, two of which are descr$bed below as methods
. 25 (A) and (B).
In method (A), the compound ls treated
successively with:
(1) a deprotecting agent, for example, an
lnorganic acid such as hydrochlorlc acld, to
convert the -NH-Ac group to a free amlno
group:
(2) a reduclng agent, for example, hydrazlne ln
the presence of a reductlon catalyst such as
~, r ~ ~ y ; . ~
(3) a cycllzlng agent, such as HC(OCH3)3,
C~,C(OCH3)3 or CN~r, to form a rlng having the
formula:
N
\ / \\
C \\
C--P
C
~; / \ /
NH
~ . ' ' ` .
`
. .
0~2tO5173 , ~ PCT/US91/06815
h ~ U
-- 24 -
wherein P is an al~yl ~roup such 48 a ~ethyl group, anamino group or hydrogen. In sethod (A), the cycllzlng
agen~ 321ec ~2~ W~ 11 dPtsr~lns ths ~dQntlty of P, For
example, if the cycllzlng agent 3elsct~d ls HC(OCH3)3, P
wlll ~e hydrcsen; ir the cyclizing agent 18 CH3C(OC~3)3,
P w~ e a ~ethyl group, and lf the cycllzlng ~gent 18
CNBr, ~ ~ the amlno. -~
In an 21t2rnatl V2 ~thcd (3), the compound
containins aa -N~-~c g oup as X-precur~or and ~n -NO2
group as Y-p-ocursor is t-eated directly wlth a metal
auch as ti-., -'no, or the ll~e, ~n the presence of
ace.ic aol~. ? ~ill th2n ~- met~yl in the r~sultlng
cyclized compound. While either method (A) or method
(~) is useful, as can be a variety of other known
cycllzing methods, cycllzlng method (~) ls preferred.
Another aspect of the inventlon relates to a
pharmaceutlcal composltlon comprl~ing a
pharmaceutlcally acceptable dlluent or carrier ln
comblnatlon wlth at least one compound accordlng to the
present lnventlon ln an amount effectlve to lnhiblt
thymidylate synthase. The composltlon preferably
contains a total ~mount of a compound of the lnvention
which ls an efficaclous amount.
The substltuted trlcycllc compounds of the present
lnventlon whlch may be amployed ln the pharmaceutlcal
composltlons of the lnventlon lnclude all of those
compounds descrlbed above, as well as the
pharmaceutlcally acceptable salts of these compounds.
Pharmaceutlcally acceptable acld addltlon salts of the
compounds of the lnventlon cont~lnlng a baslc group are
for~ed where approprlate wlth strong or moderately
strong organlc or lnorganlc aclds ln the presence of a
basic amine by meth~ds ~nown to the art. Exemplary of
the acld addltlon salts ~hlch are lncluded ln thls
lnventlon are maleate, fumarate, lactate, oxalate,
methanesulfonate, ethanesulfonate, benzenesulfonate,
: , :
-~ WO92/05173 ~ ~ L ~ PCT/US91/06815
- 25 -
tartrate, cltrate, hydrochlorlde, hydrobromlde,
sulfnte, phosphate ~nd nltrat~ s~lts. Pharmac~utlcally
acceptable baae addltlon ~lts of cc~pounds of the
lnventlon cont2ining an acldic sroup 3r~ ~rep~r~d by
known methods from organlc and lnorganlc bases and
l~clude, for eY~mple, nontoxlc al~ali ~etal and
alkallne earth bsses, such as calclum, ~odlum,
potasslum ~nd ammonlum hydro~ide; and nonto:~ic ors2nlc
b~ses such ~8 trlethylamln2, ~tyl~r.lne, piperazine,
and tri(Aydroxymethy~)me'~y?~ln-.
As stated ~ove, the compounds of the nYention
possess antlprol~feratlvP acti~lty, ~ prop2rty which
may express ltself ln the form OI ~n~icumor actlvl~y.
A compound of the lnventlon may ~e actlve per se or it
may be a pro-drug that 1~ converted ln vlvo to ~n
actlve compound. Preferred compounds of the lnventlon
are actlve ln lnhlbltlng the growth of the L1210 cell
llne, a mouse leukemla cell llne whlch can be grown ln
tlssue culture. Such compounds of the lnventlon are
al60 actlve ln lnhlbltlng the growth of bacterla such
as Escherlchla coll, ~ gram-negatlve bacterlum whlch
can be grown ln culture.
The substltuted trlcyclic compounds according to
the present lnventlon, as well as the pharmaceutlcally
acceptable salts thereof, may be lncorporated into
convenlent dosage forms such as capsules, tablets or
ln~ectable preparatlon6. Solld or liquid
pharmaceutlcally acceptable carrlers may be ~mployed.
~011~ ~;a~I _d~ 8 i .~ ~ 8t.~ Ct~ 3t C~ 1 b~
dihydrate, terra alba, sucrose, talc, gelatln, agar,
pectln, acacla, magne61um stearate and stearlc acld.
Liquid carrlers lnclude syrup, p~eanut oll, ollve oll,
; sallne ~nd water. Slmllarly, the c~rrler or dlluent
may lnclude any prolonged ralease material, such as
glyceryl monostearate or glyceryl distearate, alone or
with a wax. When a llquid carrler 18 used, the
preparatlon may be ln the form of a syrup, elixlr,
~;
W092/05173 ~ S~ PCT/US91/06815
- 26 -
cmulsion, aoft selatln capsule, ~terlle ln~ectable
llqu~d (a.g. 301utlon), such a8 an ~mpoula, or an
agueou~ or nonaqueous llquld suspanslon.
The pnar~acautioal ~rapdrationa arz ~ads ~ollow~ng
conventlonal tachnlquss of a pharmacautlcal chemlst
lnvolYing ~t3p~ 3uch as mlxin~, ~ranulating and
compressing, whPn nacessary, for t~blet forms, or
ins, filllng ~nd di solYlng the ingredlents, as
appro~-'a'o, to 91~J~ tho desired ~rcducts fo- or~l,
10 ~ar~ntaral, to~ , ir~t-;~Jaginal~ ln._2nas21,
lntrabronchi~ n.r~ocular, 'ntraaural and ractal
administration.
The compo~itions of the invention may further
comprise one or mors other compounds which are
antltumor a~ents, such as mitotic inhibitors (e.g.,
vlnblastlne), alkylating agents (e.~., cls-platln,
carboplatln and cyclophosphamlde), DHFR lnhlbltors
(e.g., ~ethotrexate, plrltrexlm or trlmetrexate),
antlmetabolltes (e.g., 5-fluorouracll and Q toslne
arablnoslde), lntercalatlng antiblotlcs (e.g.,
adrlamycln nnd bleomycln), enzymes (e.g.,
asparaglnas2), topoisomer~se lnhlbltors (e.g.,
etoposlde) or biologlcal response modlflers (e.g.,
lnterferôn).
The composltlon of the lnventlon may also comprlse
one or more other compounds lncludlng antibacterlal,
antlfungal, antlp~rasltlc, antlvlral, antlpsorlatlc and
antlcoccldlal agents. Exemplary antlbacterlal agents
- Include, for example, sulfon~mldes ouch as
sulfamethoxazole, sulfadlazlne, sulfameter or
sulfadoxlne; DHFR lnhlbltors such as trlmethoprlm,
bromodlaprim or trimetrexate, penlcilllns:
cephalosporlns: amlnoglycosides; bactarlostatlc
lnhlbltors of proteln aynthesiR; the
qulnolonecarboxyllc aclds and thelr fuRed lso~hlazolo
analogs.
..
. . .
~ u ~
~-; W092/05173 I'CT/US91/06815
- - 27 -
Another aspect of the lnventlon relates to a
therapeutic process of ~nhlbltlng thymldylats 3ynthase,
whlch proces6 comprlses admlnlstering to a vertebrate
host such a~ a mamm~l or bird an asount 3 f f3ctiv8 to
lnhlblt thymldylate ~ynthase of o trlcycllc compound
acrordlng to the present lnvent~on. The ccmpound~ of
the lnvent~on are particularly useful ln the treatment
of mammallan hosts, such as human ho8ts, and ln the
treatment of aYlan ho~ts.
Any of the substitut~d t,icy~ co~-cu-.~s
descri~ed abov~, or'pharmaceutlcally 2C-pl ab'e salts
thereof, may be employed in the th.2ra~eu .i_ ~ro~2ss 0 r
the lnventlon. The compounds of the lnventlon mcy m2
admlnistered ln the therapeutlc process of the
lnventlon ln the form of a pharmaceutlcally acceptable
compo61tlon comprlslng a dlluent or carrler, such as
those de6crlbed above. Doses of the compound6
preferably lnclude phar~aceutlcal dosage unlts
comprlslng an efflcaclou6 quantlty of actlve compound.
By an efficaciou~ quantity ls meant a quantlty
' sufficlent to lnhlblt TS and derlve the beneflclal
effects therefrom through admlnlstration of one or more
of the pharmaceutlcal dosage unlts. An exemplary dally
dosage unlt for a vertebrate host comprlses an amount
of up to about 5,000 mg of actl~e compound per square
meter of the body area of the vertebrate host.
~ he selected dose may be admlnlstered to a
warmblooded anlmal or mammal, for example a human
p~tlent, _n ne~;d of ~redtm ,~ m2~ t~;~midyla'~
synthase lnhlbltlon by any known method of
admlnlstratlon, lncludlng toplcally (e.g., as an
olntment or cream), orally, rectally (e.g., as a
supposltory), parentally, by ln~ectlon or contlnuously
by ~nfuslon, lntravaglnally, lntranasally,
lntrabronchlally, lntra-aurally or lntraGcularly.
The ~ubstltuted trlcycllc compounds accordlng to
the present lnventlon may be further cnaracterlzed as
W092/05173 ~ 3 a PCT/US91/06815
producing any one or more of an antlprollferatlve
effect, an antlb~cterlal eff~ct, an antlparasltlc
effect, an ~ntlviral æff2ct, an ntlpsorlatlc effect,
en antlproto2Gal ~f3Ctr an antic~ccldlal ~f ~3Ct or 2n
S antlfungal effect. The cc~pounds are especl~lly useful
ln produc~ng ~n antltumor effect ln a Yertebrata host
harborins a t~mor.
ThP cs~pour.ds o-^ tho pre3ent ~nvention ~re
antagonlst3 of 2 foiate co~actor and therPfora may
affect one or ~o-o otner folate-de~endent enzvmatlc
systems as well. ~mpl-s of oLhPr ,olaLe-de~:endent
enzymatic sys~ms wAich ~y ~e afec~2d includ2 5,10-
~ethyleneletr~Aydroroi~te reduc~as2, aer.n,2
hydroxymethyltransferase, and slycinea~.inerlbotide
transformylase.
The followlng examples lllustrate the lnvention,
; although the scope and ~plrlt of the lnventlon are not
llmlted thereto.
.
~'` "' ~ :, ~' ,
'
W092/05173 ~ PCT/US91/06815
- 29 -
EXAMPLES
The structures of all compound6 of the ~nventlon
were conflrmed by proton magnetlc resonanc~
spectroscopy, lnfrared spectroscopy, elemental
~lcroanalyRls or, in certaln cases, by ~aO0
spectrometry.
Proton magnetic reson~nce spectra w~ro dat_rminod
uslng a General Electrlc QE-300 6pectrcmeter 0~2L 3 ~ ns
at a fleld strength of ~OOMHz. Chemlcal ~hlfta a e
reported ln partO per mllllon (6) and ~y o2tl lr.g t~e
references such that, ln CDC13, the CHCl3 peak ls at
7.26 ppm and, ln D6DMSO, the DMS0 peak lS a- 2.49 ppm.
Standard and peak multlpllcitles are deslgnated as
followO: s, slnglet; d, doublet, dd, doublet of
doublets: t, trlplet; br6, br,oad slnglet; brd, broad
doublet; br, broad slgn~ nd m, multiplet.
Mas6 spectra were determlned using a VG 7070~-HF
high re~olution mas6 spectrometer u61ng the dlrect
lnsertlon method, an ionizing voltage of 70eV, and an
ion source temperature of 200-C. ~nfrared absorptlon
spectra were taken on a Perkin-Elmer 457 spectrometer.
~lemental mlcroanalysls gave results for the elements
stated wlth ~0.4% of the theoretlcal values.
N-N-Dlmethylformamlde (~DMF~) was dried over
actlvatod (250-) 3-A molocular ~leves, and N,N-
dlmethylacetamlde (~DMA~) tAldrich Gold Label srade)
Wa8 ~lmil~~ly drled. Tqt-ahydrefuran ~THF~) wa~
dlstilled from ~odlum benzophenone ~etyl unner
nitrogen. The term ~ether~ referO to diethyl ether.
The term ~petrol~ refers to petroleum ether of b.p. 36-
, 53~C.
Flash chromatography was performed using Sllica
~t gel 60 (Merck Art 93~5)- Where the crude solld was
insoluble in the chosen eluant, lt was dlssolved ln a
more polar solvent, and Merck Art 7734 sillca was
added. The ~lurry was evaporated to dryness on
'` ' ~
WO92/05173 PCT/US91/06815 -~
~".,1~9~
_ 30 _
rotary evaporator fltted wlth a course ~la88 frlt to
prevent spraylng of th~ 8illca. Th~ coated slllca was
then applled to the column. Thln lay_~ chrcm2tcgr~phs
(~TLC~) were performed on pr3coata~ 3haet3 of 3111ca 60
F25, (Merck Art 5719). Extracts were dried ov~r
anhydrous Na2SO~ or MgSO,. MPltins points wer~
determlned on a Mel-Temp apparatus and are uncorrected.
ExamDle 1: PreDarztlon OI Com~oun~s 2 and 3
Compcunds 2 and 3 wers prs~rad ~cco_dlns to th~
following rsaction ac~ .9
~ 10--C-`C 2~
2 ~C~C~
,' e ~ 2 h
H2,Pd(C)
THF
Preparation of Compound 2 -
6-Nltrotetrahydroquinollne
N-acyl tetrahydroquinoline, obtained by the
acylatlon (acetlc anhydride, pyrldine) of
tetrahydroqulnollne, was nltrated and deprotected as
~; descrlbed by Amlt et al., J. Chem Soc. Perklns II, 57
(1976). lH NMR (CDCl3) ~ 7.86-7.90 (2H, m), 6.36
~ 30 (lH, d, J~9.6Hz), 4.75 (lH, brs), 3.41
-` (2H, t, J~5.6Hz), 2.79 (2H, t, J~6.3Hz), and 1.91-1.99
~2H, m).
Preparatlon of Compound 3 -
` 6-A~notatrahydroquinollr.o
; 35 6-Nitrotetrahydroquinolin~, 2 (29.00g, 0.16 mol),
ln 200 ml MeOH and 10% palladlum-on-charcoal (3.00g)
was shaken on a Parr hydrogenator wlth 35 psi H2 for 1.5
.,
SUBS~T~T' SH E T
:
,~ WO92/05173 ~ 9 ~ PCT/US91/06815
- 31 -
hours. The mlxture W8S flltered through ~ dlato~aceous
earth materlal sold under the trade n~me Cellts,
concentrated, and purlfied by flash chromato~raphy (50%
- 75~ EtOAc/Hexane) to glve a yellow-brown ~olld,
15.79g (0.11 ~ol, 68~), m.p. 75-80-C. lH NMR (CDC13)
6.40 (3H, m), 3.23 (2H, t, J-5.4Hz), 2.69
(2H, t, J~6.5Hz), ~nd 1.89-1.92 (2H, ~). IR (XBr)
3400, 336~, 3900, 880, 810.
Example 2: Alternate Synthesls of Com~ound 3
~N~ H2'PtO2'AC~ ~21~.
6-Aminotetrahydroquinol~ne - Alternative Method
6-Aminoqulnoline (1.00g, 6.93 mmol) ln 20 ml
glaclal acetic acid over PtO2 (O.06g, 0.30 mmol) was
shaken at 45 psl for two hours ln accordance wlth the
procedure of Ishlkawa et al., Chem. Pharm. Bull., 37,
2103 (1989). The mlxture was flltered though a
dlatomaceous earth materlal sold under the trade name
Cellte, baslfled wlth 6N NaOH, and extracted wlth CH2Cl2
(2 x 100 ml). The organlc layer was washed with water,
drled over anhydrous Na2S0~, and concentrated to a grey
solld, 0.62g (4.18 ~mol, 60~).
Example 3: Preparatlon of Compound~ 5 and 6
c
\. p~f~ic~ c
~, 3 H H
.,~ /
~,. /
HNo3/H2So,
C ~ 10C
HN
o H
SUE~STITU T E SHE~
.
WO92/05173~iiJ~ ~ ') PCT/US91/06815
- - 32 -
Psop~r~tlo~ of C~ou~d 6 -
6-Acyl~--lno-7--Alt--ot~tr:lhy~ o~Ul301iDo
The prlmary ~mlno sroup of Compound 3 '~a3
selectlvely protected as the acet~mlde, Compound 5
(ac~tlc anhydrlde, pyridlne, -10-C).
A solutlon of Compound 5 (3.72g, 19.55 ~mol) ln 30
ml 98~ H2SO, t -10-C was trs~t2d wit~ 70~ X~03 ( 1 . ~0
~ ml, 20.22 ~mol), durlng whlch th~ ~mper-~tu-o ~nc_oased
10 to 5-C. The mixture w~ ~our~ ,00 s.l ~2~a-,
neutralized wlth 6N ~aOH, ~nd ex~r~ct~d with ~thyl
acetate (3 ~ 500 ~1). lH ~ o- t:s_ c-~d~ .~a~
showed a 2.5:1 mlxture of Compound 3 And its 5-nltro
lsomer. The extract was drled over anhydrous Na~SO~,
concentrated, and purlfled by flash chrcmatoqraphy (65%
EtOAc/Hexane) to glve 2.58g of Compound 6 as a dark red
solld (10.97 mmol, 56%), m.p. 150-155-C. ~H NMR (CDCl3)
9.80 (lH, brs), 8.22 (lH, s), 7.22 (lH, s), 4.10
(lH, brs), 3.32. (2H, t, J-5.5Hz), 2.82
20 (2H, t, J-6.4Hz), 2.22 (3H, 6), and 1.91-1.95 (2H, m).
IR (KBr) 3410, 3320, 2930, 1650, 1580, 850. HRMS,
.. exact mass calculated for C~H3N303: ~+ requ~res
; 235.0957. Found: 235.0950.
; 25 Example 4: Preparatlon of ComPounds 13 through 16 and
;, . Compounds 7(d) and 7(e)
Compounds 13-16 and Compounds 7(d) and 7(e) were
prepared aocordlng to the followlng reactlon scheme:
,
.
;:t
..
- W092/05173 ~t~J 1 6~ PCI/US91/06815
- -- 33 --
0 2
PhO)I\OPh t ¢~ ~ OC~O--~S~2--~--Ct2
CH3 1 3
/1 ) KOH I EtOH
8~:C
~/ 2~ HC
Ho~ SO2 ~Ct~;
PhC(O)CI, pyrialna / 1 ~ CH31, K2C03.
CHCi3.23C ~ \ acetnr.e
~ o~SO2~CH3
CH30~S02 ~CH3
\ 16
NBS CCI~ h~, /
'' ~ 77C
; RO ~ S0.--~CH~
7d: R-c ~o) ph
7d e 7e~ ~t~cEt3
~.~
SUBSTITUTE SHEET
!
,
W092/05l73 ~ 6 ~ ~ PCT/US91/06815
- 34 -
Prepar~tlon of Coupound 13 -
D~-4-(p-tolueneJulfonyl)ph~nyl c~rkon~t~
A solutlon of dlphenyl cJrbon~te (50.1~g, 0.23
~ol~, p-toluene sulfonyl chlorlde (90.0~g, 0.~7 ~ol)
and FeCl3 (1.12g, 0.01 ~ol), ln 75 ml nltrobenzene was
heatad to 120-C, while evolvlng HCl w~ b~blsd through
a w~ter trap. After two hours, the ~l~ture was ccol_d
to 23-C, formlng ~ llght belge preclpitate. Th2 3011d
was filter2d and w~hed wl th ~eO~ ~o g~v~ ~7.61g --udg
Compound 13 (0.17 mol, 72~ .p. l~-ioO'C.
(C~Cl3) ~ 7.98 (2H, d, J-8.7H7), 7.8~ (2~, d, J-o.3'~
7.40 (2H, ~), 7.2~-7.32 (2H, ~), and 2.40 (3r., ~).
Preparation of Cocpound 1~
~-(p-Toluene~ulfonyl)phenol
A solutlon of Compound 13 (87.61g, 0.17 ~ol3 ~n
; 150 EtOH/150 ml 5N XOH solution was heated to 80-C for
one hour. The mlxture was poured into 500 ml MeOH and
neutrallzed wlth 6N HCl, causlng preclpltatlon of the
KCl. The calt was flltered and the llquld dlssolved in
CH2Cl2, washed with water, drled over anhydrous ~a2SO4,
end concentrated to an off-whlte solld. Yleld: 69.65g
(0.28 mol, 82%), m.p. 143-144-C. lH N~R (CDC~3) ~ 7.76
(4H, d, J-8.6Hz), 7.27 (2H, d, J-8.4Hz), 6.90
(2H, d, J~8.8Hz), 6.47 (lH, brs), and 2.38 (3H, 8). IR
25 (RBr) 3325, 1900, 1200, 830, 800. Anal. calculated for
; C"Hl2O,S requlres: C, 62.89; H, 4.87; S, 12~91. Found:
C, 62.90; H, 4.91, S, 12.93.
Prep~r~tion of Co~pound 15
4-(p-Soluene-ulfonyl)ph~nyl ben-oate
A solutlon of Compound 14 (7.64g, 30.77 nmol) ln
12 ml pyrldlne/30 ml CHCl, at 23'C was treated wlth
benzoyl c~lorlde (4.20 ml, 36.18 mmol). Aftar one
hour, the mlxture was dlluted in CHCl3, w~shed with
water, drlod over anhydrous Na2S0" and concantrat2d.
~he pale belge solld was azeotroped with toluene to
glve 9.22g product (26.26 mmol, 85%), whlch was used
~. . . ;-
' . , ~ .'
.
W092/0~173 ~ S~ PCT/US91/06815
- 35 -
wlthout purlflcatlon, m.p. 192-198-C. ~H NMR (CDC13~ 6
8.17 (2H, m), 8.01 (2H, d, J~8.BHz), 7.B3
(2H, d, ~8.3Hz~, 7.66 (lH, m), 7.52 (2H, t, J~7.7Hz),
7.30-7.37 (4H, m), and 2.41 (3H, 8). I~ (X~r~ 1745,
1200, 1045, 820, 730, 700.
Pr~parat~on of Co~pou3d 16
4-(p-~oluene~ulfonyl~phenyl ethyl ether
~ m~xture of Compound 14 (20.13g, 81.07 mmol~,
X2CO3 ('~.615, 120.1B ~mol), and CH3~ (6.20 ml, 99.58
~mol) i~ 500 ml acetone was reflux2d for 4 hours. (An
~ddltlonal 1.00 ml CH3I (16.00 ~mol) was re~uire~ to
drive the reaction to completion.) The mixture was
flltered through a diatonaceous earth materlal ~old
under the name Celite ~nd concentrated. The yellow-
white solid was dissolved ln CHCl3, washed with H,0,then with brlne, and drled over anhydrous Na~S0~. The
volume was reduced, and the compound was trlturated
wlth hexane and flltered to glve Compound 16 as a whlte
solld, ll9.S3g (92%). lH NMR (CDC13) ~ 7.85 (2H, d,
J-8.9Hz), 7.79 (2H, d, J~8.3Hz), 7.27 (2H, d, J~8.4Hz),
'6.95 (2H, d, J-8.9Hz), 3.83 (3H, 8), and 2.38 (3H, s).
~Prepar~tlon of Compound 7(d)
;4~ en~oyloxyphenyl)~ulfonylben~yl bro~lde
A su6pen~10n of 4-(p-toluenesulfonyl)phenyl
benzoate (7.43g, 21.08 mmol) and N-bromosucclnlmlde
(3.76g, 21.13 mmol) ln 150 ml CCl, was heated to reflux
under a 200W llght. After one hour, lH NMR of an
allquot showed approxlmately 53% deslred product, along
wit~ 3v~ Cl~r~ic2 a.~ at~rtlng ~atarlai. The
mlxture was cocled, dlluted ln CH2Cl" washed wlth
water, drled (anhydrous Na2S0,), and concentrated to a
yellow-whlte solld, 8.36g (approxlmately 4.35g 7(d),
48%). ~he product was used wlthout purlflcatlon. lH
NMR (CDCl~) ~ 8.19 (2H, m), 7.90-8.05 (4H, 8), 7.70
(lH, ~), 7.50-7.56 (4H, m), 7.39 (2H, m), and 4.49
(2H, 8).
WO 92/05173 ! PCT/~S91/06XlS
uJ 1 ~ ~
- 36 -
Preparatlon of Co~pound 7(e)
4-(4'-~ethoxyphenyl)-ulfonylben~yl brc lde
Compound 16 was brominated as descr~ed for
Compound 7(d) to glve Compound 7(e) ln 64~ yi~ld. Tbe
product was used wlthout purificatlon. ~H NMR (CDC13)
7.86 (4H, m), 7.48 (2H, s, J~8.4Hz), 6.95
(2H, d, J-8.9Hz), 4.44 (2H, 8), and 3.83 (3H, 8).
Exam~le 5: Preparatlon of Compounds 8(a) - 8(e)
10
DMP`//Pr2NE-, c
H N ~ D~A / CaC03, 8 0 C
o2r~N ArCH;~ a-e) 2N N
6 H 8
~:A~ ~ ~S02 - h N-B~
20 b: Ar= ~S02~
C~ Ar = ~X OS~?~.2~ _
25 d.~= ~ SO ~ o ~
~ Ar~ ~ so, ~ OcH3
~:;
.Preparation of Compound 8(a) -
6-Acyla~lno-7-nltro-N-~-(N,N(l-tert-
butylcarboxa~yl)piperaz~nyl 8ul fa~oylbensyl~-
35tetrahydroqu~nollne
A solution of Compound 6 (0.277g, 1.18 mmol) and
bromide 7(a) (O.56g, 1.35 mmol) in 10 ml DMA with dry
5U85TITUTE SHEET
.
- WO92/05173 PCT/US91/06815
CaCO3 (0.1849, 1.84 mmol) wa8 heated to 80-C for 6
hours. The mi~ture w~s diluted in ethyl acet~te,
washe~ twlce with water, dried over anbydrous Na25O~,
and conc2nt_3tad. Purific~t~on by flash chro~atography
(50~ EtOAc/Hexane) g~ve Compound B(a) as a purple
801id, 0.403g (0.70 ~mol, 60%), m.p. 190-195-C. lH NMR
(CDC13) ~ 9.95 (lH, brs), 8.29 (lH, 8), 7.71
(2H, d, J~8.3Hz), 7.39 ~2H, d, J~8.2Hz), 7.12 (lH, 8),
4.57 (2~, s), 3.50 (4H, m), 3.40 (2H, m), 2.98 (4H, m),
2.90 ( H, m~, 2.2 (3H, s), 2.04 (2H, m) and 1.40
(9H, ~ ~Br) '~80, 1510, 1330. H~MS, exact mass
calc~_la~2d f_r C2.Y.3s~50,5: ~+ requlred: 573.2257.
3.2257
Preparation of Co~pound 8(b)
~-Acyla~no-7-nltro-N-~4-
(phenyl~ulfonyl)ben~yl]tetrahydroquinollne
A solutlon of Compound 6 (0.402g, 1.17 mmol),
bromlde 7(b) (0.765g, 2.46 mmol) and
dllsopropylethylamlne (O.44 ml, 2.53 ~mol) ln 4 ml DMF
was heated to 70-C for 5 hours. Workup and
purlflcatlon were as descrlbed for 8(a) to provlde 8(b)
as a red-orange solld, 0.5779 (73~), m.p. 202-205-C.
H N~R (CDCll) ~ 9.90 (lH, brs), 8.28 (lH, s), 7.89-
7.95 (4H, m, 7.50-7.52 (3H, m), 7.35 (2H, d, J-8.2Hz),
7.07 (lH, s), 4.52 (2H, 6), 3.36 (2H, m), 2.87 (2H, m),
2.22 (3H, 8 ), and 2.04 (2H, m). HRMS, exact mass
calculated for C2,H2,N,OsS: M~ requlres 465.1358. Found:
465.1373.
Prepar-tion of Co pound 8(c) -
6-Acylamino-7-nltro-N-~6-tert-butyldlpbenyl-iloxy~ethyl)
-2-naphthoben~yl]tetrahydroqulnol~ne
Compound 8(c) wa6 prep~red from Compounds 6 and
7(c) as descrlbed for Compound 8(b) wlth a 91% yleld.
~H NMR (C~Cl~) ~ 9.90 (lH, 6), 8.28 (lH, 8), 7.70-7.82
(8H, m), 7.63 (lH, ~), 7.35-7.43 (7H, m), 7.30 (lH, s),
4.91 (2H, 8), 4.65 (2H, s), 3.45 (2H, ~), 2.90 (2H, m),
2.22 (3H, 8), 2.05 (2H, m), and 1.11 (9H, 8). IR
WO92/05173 ~ PCT/US91/06815 --
- 38 -
~neat) 3360, 1680, 1080 (broad), 8B0, 820, 750, 700.
~RMS, exact m~86 calculated for C,9H"N~O,Sl: M+
requlres 643.2866. ~ound: 643.2822
Psspar~tlon of Cc pound 8(d) -
6-Acyla lno-7-nltso-N-~4-(4'-
~en~oylo~yphenyl~ulfonyl)ben~yl]-
. tetrahydroqu~nol~ne
Compound 8(d) was prepared from Ccmpounds 6 and
7(d) as de~c,l~ed for Compound 8(b) with a 57% y~eld,
~.p. 17~-17&. 1~ NMR (CDC13) ~ 9.g5 (1~, s), 8.29
(lH, s), 6.16 (2H, d, J-8.3Hz), 8.01 t2H, d, J-8.7Hz),
7.92 (2H, d, J~8.3~z), 7.65 (lH, ~), 7.51 (2H, m), 7.37
(4H, dd, J~8.6, 2.1Hz), 7.12 (lH, 8), 4.54 (2H, s),
3.36 (2H, ~), 2.88 (2H, m), 2.22 (3H, 8), and 2.04
(2H, m). IR (KBr) 3360, 1730, 1670, 1570, 1150, 880,
830, 810, 700. HRMS, exact mass calculated for
-~ C3,H27N3O7S: M+ requlres 585.1570. Found: 585.1567.
Prepar~tlon of Co~pound 8(e) -
6-acy~ no-7-nltro-N-l4-(~-~ethoxyphenyl~ulfonyl)
ben~yl]tetr~hydroqu$nol~ne
Compound 8(e) was prepared from 6 and 7(e) ln 82%
yield a6 described for 8(b), m.p. 168-171-C. lH NMR
1 (CDC13) ~ 8.30 (lH, ), 7.86 (4H, d, J-8.9Hz), 7.33
(2H, d, J-8.4Hz), 7.08 (lH, 8), 6.96 (2H, d, J-8.9Hz),
.1 25 4.52 (2H, s), 3.83 (3H, 8), 3.37 (2H, ~), 2.87 (2H, m),
2.22 (3H, s), and 2.01 (2H, m). Analytlcal data
calculated for C2sH2sN306S requlres: C, 0.59; H, 5.09;
N, 8.48. Found: C, 60.51, H, 5.12; N, 8.37.
ExamDle 5: Preparatlon of Compounds 9(f) - 9(i) and
Compounds lO(f) - lO(i)
Compounds 9(f) - 9(J) and lO(f) - 10(~) were
prepared accordlng to the followlng react~on scheme:
3S
-W092/05173 PCT/US91/06815
- 39 -
Hl ~ 3N HCI, 80C ~ ~
~Ar i Ar
Raley N! I H2NNH2
,f~ MeOH
t~2
h2N
` tO1-
,
1 5 SO?--
9 A~ = ~SO
rn: ~r~ ~_ Oh
~ 20 1 Ar= ~SO?~OI'
~ I A~= ~SO ~Octi;
~'
Prepar~tlon of Compounds 9(f) - 9(~) -
- 6-Anlno-7-nltro-N-~-(N,N-plperazlnyl~ulfa~oyl)benzyl]-
tetrahydrogulnollne, 6-A~lno-N-t~(phenylsulfonyl)-
benzyl~totrahydroqulnollne, 6-A lno-7-nltro-N-t~6-
hydroxymethyl)-2-naphthobenzyl~tetrahydroqulnollne,
6-A~lno-7-nltro-N-~-(4'-hydroxyphenyl-ulfonyl)-
benzyl~tetrahydroquinoline, und
6-Aolno-7-nltro-N-t4-(~'-
ethoxyphenyl 8ul fonyl)benzyl]tetrahydroqulnollne
~he followlng procedure, descrlbed for Compound
9(f), is representative A suspenslon of Compound 8(a)
(0 40369, 0 70 mmol) in 3N HCl (12 ml) was heated to
SV85TITuTE SHEET
W092/05l73 PCT/US91/06815
v ~
- 40 -
60-C for 5.5 hours. ~he m~xture w~s poured ~nto 100 ml
~aturatsd NaHCO~, extracted 4 tl~es w~th ethyl acstate,
drled over anhydrous Na2S0" and concentrated.
PurlIlca~lon by flash chromatography (10~ EtOH/CHCl3)
ylelded 0.2159 Compound 9(f) as ~ purple solld (0.50
smol, 71~), m.p. 194-196-C. ~H N~R (CDCl3) ~ 7.71
(2H, d, J-8.2Hz), 7.42 (2H, d, J~8.1Hz), 7.06 (lH, 6),
6.52 (lH, s), 5.67 (2H, brs), 4.50 (2H, s), 3.31
(2H, t, J~5.7H_), 2.95-2.99 (4H, ~), 2.90-2.93 (4H, m),
2.80 (2~, t, J~S.0Hz), and 2.01-2.03 (2H, m). IR (~3r)
34~0, 3420, 12~0. HRMS, exact ~ass calculatad for
C20~Z3~3la: ~t requlres 431.1627. Found: 431.lS26.
9(g): lH NMP~ (CDC13) ~ 7.B8-7.94 (4H, m), 7.50-7.54
(3H, m), 7.38 (2H, d, J-8.3Hz), 7.02 (lH, s), 6.50
(lH, 8), 5.65 (2H, brs), 4.46 (2H, s), 3.25-3.27
; (2H, m), 2.75-2.77 (2H, m), and 1.90-2.04 (2H,m).
,
9(h): lH MMR (CDC13) ~ 7.79 (3H, ~), 7.68 (lH, s),
7.39-7.48 (2H, m), 7.25 (lH, s), 6.51 (lH, s), 5.63
(2H, brs), 4.85 (2H, d, J-5.9Hz), 4.58 (2H, 8), 3.33
' (2~, ~, J~5.7Hz), 2.79 (2H, t, J~6.3Hz), 2.04 (2H, m),
~nd 1.71 (lH, t, J~6.0Hz). IR (XBr) 3480, 3320, 2910,
1560, 880, 810. HRMS, exact mass calculated for
C2,H2,N503: M+ requlres 363.1583. Found: 343.1574.
9(1): ~.p. 200-250-C: ~H N~R (CDCl3) ~ 7.80-7.86
(4H, m), 7.37 (2H, d, J~8.2Hz), 7.01 (lH, 6), 6.90
(2H, d, J~B.7~z), 6.50 (lH, 8), 5.66 (2H, brs~, 4.46
(2H, 8), 3.28 (2H, t, H~5.7Hz), 2.77 (2H, t, J-6.2Hz),
and 1.99 ( 2H, m). IR (KBr) 3460, 3380, 2940, 1570,
1150, 880, 840. HRMS, ~xact mass calculatod for
C22H2lN305S: M+ requlres 439.1202. Found: ~39.1213.
9(J): m.p. 141-143'C: lH NMR (C~Cl3) ~ 7.86
(4H, d, J~8.9Hz), 7.35 (2H, d, J-8.3Hz), 7.03 (lH, s),
6.96 (2H, d, J-8.9Hz), 6.50 (lH, ~), 5.65 (2H, brs),
WO 92/05173 ~ 6 ~ ~ PCT/US9l/06815
4.~5 (2H, 8), 3.83 (3H, 8), 3.26 (2H, t, J~5.BHz), 2.77
(2H, ~), 1.98 (2H, ~). I~ (K;3r) 3400, 1500, 1250,
1150, 1100, 860, aoo.
~_ ~p~sation of ~ou;~ 10 ( f ) -10 ( ~ ) -
6, 7 -D ~no-M- [ ~ , N -p lp~r~z lslyl ~ul f asoyl ) b~ansyl ~ -
-2t~ ydsc5;u 1 ~ol1~, ~, 7-Di~al~o-?l~
(ph~nylalulfonyl)3~syl] -
t~ts~hydr~ui~oli~, 6, 7-D1~3ino-~-
r ~ 5-hydrc:s~ethyl ) -~-
10 ;:ap;~ c~æ~~yl~t~,.r~hy~ul.. oline,
6, 7 -3i 3~ir.0-~- i , -4 ' -~ydro3~ ~hæ yl3ul f or.yl ~ -
~zyl ] t~trahydr~uinoli~2, and
6,7-Diamino-~;-[~-~'-~ætnoxyphanyl~ulfonyl)-
~en~yl~tetrahydroqu~nollne
The followiny procedure described for Compound
lO(f) is r~present~tlve: A solution of Compound 9(f)
(O.215g, 0.50 ~mol) ln 6 ml MeOH/2 ml THF over ~
catalytlc ~mount of Raney Ni/H20 was heated to reflux
and tre~ted w~th an~ydrous ~ydrazlne (1.00 ml, 31.54
; 20 ~mol). After one ~our, durlng whlch loss of coior
oscurred, the mixture was filtered through a
dlatomaceous earth material sold under the trade name
Celite, concentrated, and azeotroped wlth CH3CN/~enzene
to glve 0.158g of Compound lO(a) (0.39 ~mol, 79~). In
all ca6es, the dlamlne was used lmmedl~tely for the
cycllzation reactlon. IH NMR (C~Cl~) ~ 7.69
(2H, d, J~8.2Hz), 7.44 (2H, d, J~8.2Hz), 6.44 (lH, 8),
S.85 ~lH, ~), 4.42 ~2H, 8), 3.24 (2H, m), 2.97 (4H, m),
2.93 (4H, m), 2.70 ~2H, m), and 1.98 ~2H, m).
Example 6: Preparatlon of Com~ound 11 and Compounds
12(f)-12(i) of the Inventlon
Preparatlon of Co~ound 11 -
6-7-Im~da~ol-N-l~-(N,N-pipera~lnyl~ulfamoyl)-
ben~yl~tetrahydroqulnol~na
WO92/05173 PCT/US91/0681S
- 42 -
~`~ HC(OM3)~, HCI
10t ~ ~ H11
PIlSC)2 PhSO2
.~ I I
N N
H
.~ soluz~o~ o- diamine Compound 10(f) (0.084 g,
0.21 ~mol) in 2 ml HC(OC~3)3 at 23C was t.2ated with
one drop of concentrated HCl, causlng the formatlon of
a precipitate. The mixture was stirred for one hour,
poured into saturated NaHCO3 ~olution (20 ml), and
extracted wlth 50 ml CHCll. The organ~c layer was dried
wlth anhydrous Na2SO4, concentrated, and purified by
flash chromatography (1.5% NH3/13.5% EtOH/85~ CHCl3) to
glve Compound 11 as a yellow-orange solld ~0.032g, 0.08
mmol, 38%). lH NMR (acetone-d6) ~ 8.01 (lH, s), 7.76
(lH, s), 7.70 (2H, d, J-8.1Hz), 7.56 (2H, d, J-8.1Hz),
7.20 (lH, s), 6.52 (lH, s), 4.64 (2H, s), 3.45
(2~, t, J-5.7Hz), 2.92 (2H, t, J-6.1Hz), 2.84
(4H, brs), 2.81 (4H, brs), ~nd 2.04 (2H, m). HRMS,
exact mass calculated for C2~H2sN5O2S: M+ requlres
411.1729. Found: 411.1719.
Preparat~on of Co~pounds 12(f) - 12(~) -
6,7-(2-A~inolmidazol) -N- t ~- ( N, N-
piperazlnyl-ulfa oyl)benzyl~totrahydroquinol$ne,
6,7-(2-~minol~idazol)-N-[~-(phenyl~ulfonyl)benzyl]-
; tetrahydroqu~noline, 6,7-(2-A~inol~$dazol)-N-[(6-
hydroxymethyl)-2-
naphthobenzyl]tetrahydroquinoline,
$,7-(2-Xzinoi~idazol)-N-[~-(4'-
hydroxyphenyl 3ul fonyl)~enzyl]t2trahydroquinoline, and
6,7-(2-A~noi~idazol)-N-t4-(4'-
nethoxyphenyl~ulfonyl)benzyl]tetrahydroquinollne
c~ l~c~ SY.ET
:' . .
.
. ~ WO 92/0~173 ~ v ~ 3 ~ PCr/l~S91/06815
- 43 -
H2N~ CN~~ ~eO
lOt-l ~ H 12t-
Ar
. ~ .
The followin~ procodu-o, dosc~ d for 12(i), is
representative: A soiu~ion o: diamin2 Compound 10(i)
(1.59 mmol) ~n 6 ml CH3C~'~2 r,i Me3~ was haate~ to 70C
and treated with 5. Or C~Br~Cn3CN solution (0.5 ml, 2.5
mmol). After two hours, the mixture was diluted in
0.5N HCl and extracted twice wlth ethyl acetate. The
aqueous layer was neutralized with 6N NaOH, saturated
wlth NaCl, and extracted twice with ethyl acetate. The
organic layer was dried over anhydrous Na2SO~ and
concentrated to a brown solld. Puriflcatlon by flash
chromatography (5% to 10% EtOH/CH2Cl2) gave Compound
12(1) as a llght brown solld, 0.173g (0.40 mmol, 25%),
m.p. 230-245C. lH NMR (DMSO-d6) ~ 7.86 (3H, m), 7.75
(2H, d, J-8.7Hz), 7.43 (2H, d, J-8.2Hz), 6.91 (3H, m),
6.16 (lH, s), 4.53 (2H, s), 3.35 (2H, m), 2.80 (2H, m)
and 1.90 (2H, m). IR (KBr) 3400 (broad), 1660, 1280.
HRMS, exact mass calculated for ~23H22N~O3S: M+ requires
434.1412. Found: 434.1401. ~3C NMR (DMSO-d6) ~ 162.3,
149.5, 144.8, 141.5, 140.7, 130.7, 129.7, 129.2, 127.5,
127.4, 127.2, 120.3, 118.0, 116.2, 111.5, 93.3, 54.7,
49.9, and 28.3.
12(f): ~H NMR (CDCl~) ~ 7.65 (2H, d, J-8.3Hz), 7.48
(2H, d, J~8.2Hz), 6.93 (lH, s), 6.10 (lH, s), 4.52
(2H, s), 3.43 (2H, t, J~5.5Hz), 3.29 (4H, ~), 3.04
(4H, m), 2.92 (2H, t, J~6.5Hz), and 2.17 (2H, m). I~
(K~r) 3420, 1610. HRMS, exac~ mass calculated for
C2~H26N6O2S: M+ requires 426.1833. Found: 427.1940.
SUBSTJ7'UT~ SLI.EET
, ' .
', " ' "'', .; ' ' ' ' ' ,. ' " ~. ` ,." ,. , .,, ', '. . ';: , ', '
.
W092/OS173 r~ '~ iJ` i ~ QJ ~ PCT/US91/06815
- 44 -
12(~ H ~MR (CDC13) ~ 7.92 (2H, d, J~B.3Hz), 7.83
(2H, d, J~8.3Hz), 7.47-7.55 (3H, ~), 7.39 (2H, ~), 6.8g
(lH, ~), 6.23 (lH, 8), 4.44 (2H, 8), 3.32 (2H, ~), 2.85
(2H, ~), and 2.~1 (2~ R (f~l~) 1630, 1290, 1150,
820. ~RMS, exact ~ass calcul~tad for C23H22N402S:
M+ roquirsd ~18.14~3. Found: 418.1487.
- 12(h)~ ~SO-ds) 4 7-71-7-3~ (4~, ~), 7-~1
(2~, ~), ~.5a (1.~, 8), ~.30 (lH, 8), 5.74 (2H, brs),
10 ~-~2 (~ J ~ , O ), 3.3~ , ~), 2.77 (2~, ~),
and 1.93 ~X, ~ ~3r~ 34~, 1720. HRMS, exact
mass calculat~d for C22'~22N,O: ~i r qulras 358.1793.
Found: 35R.181~.
.
15 12(~): ~.p. 217-228-C; lH NMR (D~SO-d6) ~ 7.84
(4H, d, J-8.9Hz), 7.46 (2H, d, J-8.3Hz), 7.09
(2H, d, J-8.9Hz), 6.66 ~lH, 8), 6.10 (lH, 8), 5.74
(2H, br~), 4.45 (2H, 8), 3.80 (3H, 8), 3.15-3.26
(2H, m), 2.74 (2H, m), ~nd 1.83 (2H, m). IR (KBr)
20 3400, 2950, 1650, 1600, 1400, 1140, 1100, 830, 800.
HRMS, exact mass calculated for C2,H"N,035: M+ requires
448.15~9. Found: 448.1559.
Example 7: Preparatlon of Compound 17
6,7-(2-Methyllnldazol)-N-
[4-(4'-~etboxyphenyl~ulfonyl)ben~yl~-
tetr~bydrosu~nollno
Compound 17 was prepared by the followlng
reactlon:
~ . .
.
;
.
: .
~vl~3~)
- W092/05173 PCT/US91/06815
- 45 -
Alc
HN
"' B(e)
so2~
Sn, AcOH
renux
., 10 1 i
H3C~
H 17 ~3
SO2~
20, b~ ~ OCH3
A solution of compound 8(e) (0.1131~, 0.228 mmol)
ln 3 ml glacial acetic acid was treated with Sn
(0.1443~, 1.216 mmol) and refluxed overnisht. The
mixture was filtered through a diatomaceous earth
material sold under the trade name Celite, neutralized
wlth satùrated NaHC03 solutlon, and extracted twlce with
ethyl acetate. The combined organic layers were dried
over ~nhydrous Na2S0~, concentrated, and purlfled by
flash chromatography (10% ~eOH/CH2Cl2) to give Compound
17 as a light yellow solid, 0.0724g (0.162 mmol, 71~),
; m.p. 162-165C (decomposition). Exact mass calculated
for C2sH2,N303S: 447.1616. Found: 447.1586. IR (RBr)
3400, 1590, 1250, 1140. lH NMR (CDCl3) ~ 7.84 (4H, m),
7.38 (2H, d, J~8.3Hz), 7.16 (lH, 6), 6.96
~2H, d, J-8.9Hz), 6.33 (lH, s), 4.49 (2H, s~, 3.83
(3H, s), 3.38 (2H, m), 2.92 (2H, m), 2.47 (3H, 5), and
2.04 (2H, m).
SUESSTITUTE SHEET
. . . ,: , . , .` . . . .
:.. ...
..
~.;
W092/0~173 !~r PCT/US91/06815
v ~
- 46 -
Ex~mple 8: ~etermlnation of Inhlbltlon Constants
A~alnst 5,10-Meth~lene-tetrahYdrofolate
Thymldylate ~ynthaQe lnhibltlon constants K~ h~ve
been detar~lnad ~y th~ ~ollo~lng ~ethod. All as~ay6
were run at 25-C and lniti~ted by the sddition of three
dlfferent types of thymldylate ~ynthase (~TS~): (1)
Escherichla coll TS (~TS~ ) C~ndld~ TS (~CTS~),
whlch is rrcm 2 fung~s; and (3) hu~an TS (~TS~).
TS exhibit~ ordered blrsdctant Xlnetics (Daron,
H.H. and Aull, J.L., J. B~ol. Ch2m. ~53, 9~0-9~al
(1978), and tha d~ ~2'-d~oxyurldln2-5'-~ono?hosphate)
conc~ntratlor.~s~d fcr thas~ ,~ac ~ionO W25 nPar
saturation leveis so ~at ~Aa a~Oay~ ~e,~ p~audo-~nglP
substrate. All reaction mixtures contained 50 mM Tris
at pH 7.8 (the flnal pH of the reactlon was 7.6), 10 mM
DTT (dlthlothreitol), 1 mM EDTA
(ethylenedlamlnetetraacetlc acld), 25 mM mgCl2, 15 mM
H2C0 (formaldehyde) ~nd 25 mlcroM dUMP. When human ~S
wa6 assayed, lOO mlcrograms/ml of BSA (bovlne serum
albumln) was present ln the reactlons. The range of
THF (tetrahydrofolate) was 5 to 150 mlcroM (elght
concentratlons: 5, 6.6, 10, 13, 16, 25, 50 and 150
mlcroM). A standard curve in the ab~ence of lnhlbitor
8 run wlth each experlment. Three cur~es were then
run wlth lnhl~ltor at three different concentrat~ons
- wlth a mlnlmal range, where posslble, from 1/2 to 2
tlmes the K, (Cleland, W.W., Biochlm. Blophys. Acta 67,
173-187 (1963). ~hese assays were done on a
spectrophotometer at 340 nm (Wahba, A.J. and Frledkln,
M., J. Biol. Chem. 236, PC11-PC12 (1961), followlng the
formatlon of D~F (dlhydrorolate) (mM extlnctlon
coefflclent of 6.4) or by followlng the release of
trltlum (Lomax, M.I.S. and Greenberg, G.R., J. Blol.
Chem. 242, 109-113 (1967) from the 5-po~ltlon of dUMP
(a88ay8 for tritium relea~e cont~ln2d 0.5 mic~ocI
dUMP). Charcoal was used to remove unreacted dUMP from
the trltlum release reaction mlxtures, and the
'
. .
WO 92/05173 PCT/US91/06815
G 9 ~
- -- 47 -
~` resultant wster was counted to detennlne the extent o~
re~ctlon. Inhlbitlon constants were then deter~lned by
::~ plottlng the apparent JC~ or the reclprwal of the
apparent V.., agalnst the lnhlbltor concentr~tlon
5 (Cleland, W.W., 'rhe Enzyme~, 2, 1-65 (1970)).
~he results obtalned are t~bul3ted in T~bl~ I:
;
' : ~; ' :
.
WO92/05173 . ~ ~3 PCI/US91/06815
J
. -- 48 --
~ABLE _I
, p~
.,. U
, ` s,.
! ~ ~,
C~ d Y ~ ur~c r~c
5,7 ~ C ~ ,...... 7 r
12(f ) -NH2 ~ o2-~/\ 1111 0.2~. 0.17 0.25
1 2 ( g ) -N~2 ~50 ~(Y~ 0 . 0 6 2 0 . 0 4 8 0 . 1
.~ ~
..
,, ~
12(h) -N.~.~ ~oY C.C~l 0.074 0.086
12(i~ -NH2 ~502~r C.018 0.013 0.007
17 -CH3 t~92~- OC~ . S 1 . 7
,:,g
12~ -NH2 ~fS2~ (~ ~ ~ C.033 0.030
,;
~,
SUBST~lJTE SH~T
, ~ .
';' .,
. ..
WO92/05173 ~ ,9 ~ PCT/US91/06815
- 49 -
ExRm~le 9: In Vltro Testln~
Cellular growth ln the presence of the compounds
ln questlon was assessed usln~ three cell llne~
the L1210 murlne leukemla (ATCC CCL 219); (2) CCRF-
C~M, a ~uman lymphoblastlc leukemi~ llne of ~-cell
-~ orlgln (ATCC CCL 119); and (3) a thymldlne ~lnas2
deficlent human adenocarclnoma llne (GC3/M TX). ~oth
lines were ~alntalned ln RPMI 1640 ~edlum contalnlng 5
heat-lna~tlvated fetal bovlne serum wlthout
antlblotlcs.
ICso values were determlned ln 150 microllter
mlcrocultures each containing 1500 (L1210), 4000 ( CCRF-
CEM) or 10,000 (GC3/M TK) cells establlshed in 96 w211
plates ln growth medlum supplemented wlth 50 IU/ml
penicillin and 50 mcg/ml streptomycin. Growth was
measured over 3 days (L1210) or 5 day~ (CCRF-CEM and
GC,/M TK) of contlnuous expo~ure to varylng
concentrations of each test compound added 4 hours
after lnltlal cell plating by the MTT-tetrazollum
reductlon assay of Mosmann, T.J. Immunol. Meth., 65,
SS-63 (1983), modlfied according to Alley et al.,
Cancer Res. 48, 589-601 (1988). Water insoluble
derivatlves were dlssolved ln DMS0 and diluted as a
flnal concentratlon of 0.3% solvent ln cell cultures.
2S The results obtalned from thls procedure are
tabulated below in ~able II.
' - ,.
. :~ . . . .
' '. .
WO92/05173 PCT/US91/06815 -
h ~ ~J i ~i $ ~
-- 50 --
TABLE II
P ~
Cm~d $ p Ar L'2 ^ CC~~C~`.~ 'J~
r. ~ ~2-~ 3 n C
12(f)-~r:2 ~ ~ 2.05 2.2 >5
,
12(g) -NH2 ~2~ 2.0 3.9 >5
12(h) ~ ~ 3.1 >~ >~
OH
12(i) -NH2 ~2~ 12 22 ~50
. 17 -c~3~2~ ~C~, 2.8 8~0 ~.'
.,
12(~)NH2 ~2~ 2.~ 5.1 S.C
~','
SU13STITUTE SHEET
.
: ` ,
.
WO 92/05173 ~ ~ J i ~ èJ IJ PCT/US91/06815
While the lnvention has been descri~ed ln detall
and wlth reference to speclflc embodlment~ thereof, lt
will be app~rent to one skllled ln the art th~t ~rlous
changes and modlflcatlons can be made thersln wlthout
departing from the splrlt and scope thereof. Thus, lt
18 lntended that the present lnventlon cover the
modiflcations and variations of thls lnventlon provided
th~y come wlthin the scope of the appended cl~ims ~nd
thelr equlvalents.
