Note: Descriptions are shown in the official language in which they were submitted.
WO 92/16508 2~ PC1/l'S92tO1~6
"~
t" - ` !
REISSERT COMPOUNP~S ANTI-HIV AGE~
Field of the Invention
This invention provides novel 1,2-dihydroquinolines, 1,4-dihydroquinolines, 1,2,3,4-
tetrahydroquinolines, 1,2-dihydroquinazoliDes, and 1,2-dihydroquinoxalines, as well as known
compounds of these classes, which are useful as anti-AlDS drugs.
Baçkeround o~f~ Qc
An estimated one to one and one-half million people in the United S~ales are infected
with a human retrovirus, the human immunodeficiency virus type I (HIV-I) which is the
etiologicaJ agent of acquired imrnunodeficiency syndrome, AIDS, see Science, 661~62 (1986).
Of those infected, an estimated two hundred and fifty thousand people will develop AlDS in the
next five years, see Science, 1352-1357 (1985). OD March 20, 1987, the FDA approved the
use of the çompound, AZT (zidovudine), to treat AIDS patients with a reçent initial episode of
pneumocystis carinii pneumonia, AIDS patients with conditions other than pneumocystis çarinii
pneumonja or patients infeçted with the virus with an absolute CD4 Iymphocyte çount of less
than 200/mm3 jD tbe peripheral blood. AZT is a known inhibitor of viral reverse transcriptase,
an enzyme necessary for human immunodeficiency virus replication.
U,S. Patent 4,724,232 claims a method of treating bumans baving acquired
immunodeficiency syndrome utiliziog 3'-azido-3'-deoxy-thymidine (a~idothymidine, AZT).
It is known in the art that certain antibiot;cs and polyanionic dyes inhibit retrovirus
reverse transcriptase. It has also been reported that various sulfated compounds inhibit virus
replication, including HIV.
Information Disclosute
F.D. Popp, ~Reissert Compounds and Related N-acyldihydroquinolines~ in Quholines,
Part 11, Ed. C. Jones, (1982) p. 353-372, is a general review covering the synthesis and
reactions of Reissert compounds. This review also specifically describes the synthesis of 1-(4-
chlorobenzoyl)-2-cyano-1,2~ihydroquinoline, 1-(1 fluorobenzoyl)-2-cyano-1,2-
dihydroquinoline, l~methoxybeyl)-2~yano-1,2-dihydroquinoline, 1-(4-toluoyl~2-cyano-
1,2~ihydroquinoline, I-(~nitrobenzoyl)-2~yano-1,2-dihydroquinoline, 1-(2-naphthoyl)-2-
cyano- 1,2-dihydroquinoline, and I -(I -oxo-3-phenyl-2-propenyl)-2-cyano-1,2~ihydroquinoline.
F.D. Popp, W. Blount and P. Melvin, 1 Org. Chem., 26, 4930 (1961), describes themethylene chloride/water method for synthesizing Reissert Compounds.
S. Ruchirawat, N. Phadu~glcul, M. Chuankamnerdl~arn, and C. Thebtaranonth,
~eterocycles, 6, No. 1, 43 ~1977), describes the trimethylsilyl cyanide med~od of synthesiziDg
Reissert compounds.
M. Rozwadowska, Roczniki Chemii. 51, 2321 (1977), describes hydrogenation of
- SUBSTITUTE SHEET
~: .- ~ . ~ : . :
, . . . .. . . . .
, . . . .
~vo 92J16508 PC-r/US92/017~6
2~ ~133~ 2- ~
dihydroquinoline Reissert compounds to tetrahydroquinolines.
L. Wal~ers, M. Siegel, and R. Cook, I HeterQcyclic ~ , 5,577 ~1968), describes
synthesis of amidoximes via addition of hydroxylamine to a Reissert compound.
H. Bartsch, O. Schwan, and G. Neubauer, Heterocycle~, 24, 3483 (1986), describesthe synthesis of a benzoxazine Reissert compound.
F. Elroy and R. Lenaers, ~The Chemistry of Amidoximes and Related Compounds,~
~h~. ~. 155 (1962), describes synthesis and reactions of amidoximes.
Uff, B C., Budhram, R.S., Consterdine, M.P., Hicks, J.K., and Slingsby, B.P.,
J. Chem Soc., ~Ç~ ~ 1 (18): 2018-22 (1977), describes synthesis of 2-cyano-1-(4-
methoxybenzoyl) 4 methyl-1,2-dihydroquinoline.
F.D. Popp and A. Soto, I ~ oc., 1760 (1963), describes the synthesis of 1-(4-
fluorobenzoyl)-2-cyano- 1,2-dihydroquinoline.
~apanese Patent Applicatioo JO 2049-782-A, discloses quinolidine compounds which are
stated as useful n treating AlDS. lbese quinolidines are not structurally related to the
compounds of the present invention.
SUMMARY OF THE INVENTION
This invention provides compounds of formula I whereîn
R2 is -CN, -N3, -SCN, and the E or Z isomers of -C(=NORlo)NH2, -C(-NRlo)H,
-C(=NNRSo)NH2, -C(=NRlo)NH2, -C(=NORlo)H, and -C(=NOH)N(RIo)H;
Rlo is hydrogen, Cl-C4 alhyl, -Ac, -C(O)OCH3, -C(O)OC2Hs, -phenyl, -P02-0-
cation+, -CO-CH(M)NH2, -CO-C6H6-NRI IRl2, and -CO-C6H6-CH2-NRI IRl2;
AA is-CH3, -CH(CH3)2, -CH2CH(CH3)2,-CH2OH, -CH(OH)CH3, -CH2CO2H,
-cH2cH2co2H~ -(CH2)4NH2. and -(CH2)3-NH2;
R~l and R12 may be the same or different and are hydrogen and C1-C4 allcyl;
Rl l and R12 taken together are morpholinyl, piperazinyl, pyrrolidinyl, piperadinyl, and
N-(CI-C4 allcyl)piperazinyl;
R3, present when the connection between positions 2 and 3 is a single bond, is
hydrogen, Cl-C4 all~yl, -(CH2)"CN, and -(CH2)bR~3;
R13 is -N(CH3)2, -OH, -OPO3H, -OCH2CH(-OH)CH2OH, -OCH2CO2H,
-O2C(CH2)2CO2H,-OR14, and -NR14;
R14 is a N-terrninal arnino acid;
a is zero to six;
b is one to five;
A is CH, CH2, N, and CHN(CH3)2;
X is CH, CH2, NH, O, and C-R4;
R4 is-CH3 -OCH3, and -OAc;
SUBSTITUTE SHEET
~'
. ~
: ~,
WO 9~/16508 2 1 0 1 3 ~ 5 PCr/US92/ol746
3-
R5, R6, R7, and R8 may be the sarne or different and are hydrogen and nuorine;
W is o~ygen and sulfur;
Z is a dire bond or a linker selected from the group consisting of -(CO)-,
-CH=CH-CH=CH- (all E), -CR~5=CR16- (E or Z), and -CH=C=CH-;
RlS and R16 may be the sarne or different and are hydrogen and fluorine;
Yl is hydrogen, -Cl, -F, -Br, -CH3, -CF3, -(CH2)CRl7, -CHO, -CO2CH3, -OH,
-OCH3 -OAc, -CN,-NO2, -SH, and-SCH3;
c is zero to five;
R17 is-CH2N(CH3)2, -OH, -OPO3H, -CH2N(C2H5)2~ -O-sugar,
-OCH2CH(OH)CH2OH, -OCH2CO2H, -O2CCH2CH2CO2H, -CH2N = CHN(CH3)2, -ORI 8~ or
-NR~8;
R18 is an N-terminal a nino acid;
Y2 is hydrogeD, -F, -Cl, -OCH3, and -CF3;
Yl and Y2 talcen together can form -OCH20-, -OC(CH3)20-, -OCH2NH-,
-NHCH2O-, -OCH2S-, -SCH2O-, and -CH = CR19-CH = CH-;
Rlg is hydrogen, -Cl, -Br, -NO2, -CF3, -CO2CH3, -OH, -CN, and -OAc;
provided, however, when Z is a direct bond and A and X are each CH, and R5, R6,
R~, and R8 are each hydrogen, and R2 is -CN, Yl is not -Cl, -F, or -CH3 and Rl9 is not
hydrogen,
when Z is a direct bond and R2 is -C(=NOH)N(RIo)H and Rlo is hydrogen, R19 is not
hydrogen,
when Z is a direct bond, Yl is not hydrogen,
when Z is ~R~5=CR~6- (E), and Y2, R15, and R16 are each hydrogen, Yl is not
hydrogen,
when Yl is -OCH3 ~nd Z is a direct bond, at least one of Rs, R6, R7 or R8 is fluorine,
when Yl is -CF3 and Z is direct bond, the connection between positions 3 and 4 is a
double bond,
when Z is a direct bond, X is not O,
when Z is a direct bond, R2 is not -N3,
when Z is a direct bond, Y2 is hydrogen,
when A is N, X is C-R4 and R4 is -OCH3, or -OAc and the coMection between
position 3 and 4 is a double bond,
when A is CHN(CH~)2, X is CH2
when R4 is -CH3, R3 is hydrogen,
when R4 is-CH3, Yl is -Cl, -F, -Br, -CF3, ~N, or -NO2,
when R5 is fluorine, R6, R~ and R8 are each hydrogen,
SUBSTITUTE SHEET
3: ' . '- . .
-,~ ~ . ,
, ' ' ' ' ' ' . ' '' ~, ' ,' . . :
~vo 92/16508 2 1 ~ 1 3 3 5 PCr/US92/ol746
-4-
when Yl and Y2 taken toge~her are {:H=CR19-CH=CH-, Z is a direct bond, and
when Yl and Y2 taken togelher can form -OCH20-, -OC(CH3)20-, -0CH2NH-,
-NHCH20-, -OCH2S-, and -SCH20-, Z is a linker;
or the pharmaceuticaJly acceptable sal~s thereof.
S nle preferred compound is l-E-(3-(4-chlorophenyl)-1-o~o-2-propenyl)-2-cyano-1,2-
d ihydroqu inol ine.
The invention further provides compounds of formula n wherein
R2 is -CN, -SCN, and the E or Z isomers of -C(-NORI~)NH2, -C(=NRlo)H,
-C(= NNRlo)NH2, -C(= NRlo)NH2, -C(= NORIo)H, and -C(= NOH)N(RIo)H;
Rlo is hydrogen, Cl-C4 alkyl, -Ac, -C(O)OCH3, -C(O)OC2HS, -pbenyl, -PO2-O-
cation+, -CO-CH(AA)NH2, -CO-C6H6-NRlIR~2, and -CO-C6H6-CH2-NRIIR12;
AA is-CH3, -CH(CH3)2,-CH2CH(CH3)2.-CH2OH, -CH(OH)CH3, -CH2CO2H,
-cH2cH2co2H~ -(CH2)4NH2. and -(CH2)3-NH2;
Rll and R12 may be the sarne or differen~ and are hydrogen and Cl-C4 alkyl;
Rl I and R12 taken together are morpholinyl, piperazinyl, pyrrolidinyl, piperadinyl, and
N-(CI-C4 alkyl)piperazinyl;
R3, present when the conne tion between positions 2 and 3 is a single bond, is
hydrogen, Cl-C4 alkyl, -(CH2),CN, and -(CH2)bR13;
R13 is-N(CH3)2, -OH, -OPO3H, ~CH2CH(OH)CH2OH, -OCH2CO2H,
-02C(CH2)2C02H, -OR14, and ~ 4;
R14 is a N-terminal amino acid;
a is zero to six;
b is one to five;
A is CH, CH2, and CHN(CH3)2;
X is CH, CH2, and C-R4;
R4 is -CH3, -OCH3, and ~Ac;
R5, R6, R7, R8 may be the same or different and are hydrogen and fluorine;
W is o~tygen and sulfur;
Yl is-CI,-F, -Br,-CH3,-CF3,-(CH2)cRl7,-CHO,-CO2CH3,-OH,-OCH3 -OAc,
-CN, -NO2, -SH, and -SCH3;
c is zero to four;
R17 is-CH2N(CH3)2, -OH, -OPO3H, -CH2N(C2H5)2, -O-sugar,
-OCH2CH(OH~CH20H,-OCH2C02H,-02CCH2CH2C02H,-CH2N=CHN(CH3)2,-OR~8,
asld -NR~8;
R18 is a N-terrninal amino acid;
Y2 is hydrogen;
SUBSTITUTE SHEET
...
- :~
.
wo 92/16508 2 1 ~ 1 3 3 5 ` PCT/I IS92/01746
f, '.' . -S~
Yl and Y2 ta~en together are -CH=CRI9-CH=CH-;
Rl9 is hydrogen, -Cl, -Br, -NO2, -CF3, -CO2CM3, -OH, -CN, and -OAc;
provided, however, when A and X are each CH, and R5, R6, R7, and R8 are each
hydrogen, and R2 is -CN, Y~ is not ~1, -F, or -CH3 and R19 is not hydrogen,
S when R2 is -C(=NOH)N(RIo)H and Rlo is hydrogen, Rl9 is not hydrogen,
when Yl is ~CH3, at least one of R5, R6, R7 or R8 is fluorine,
when Yl is -CF3, the connection between positions 3 and 4 is a dauble bond,
when R4 is -CH3, R3 is hydrogen,
when R4 is -CH3, Yl is -Cl, -F, -Br, -CF3, -CN, or -NO2, and
î0 when R5 is fluorine, R6, R7 and R8 are each hydrogen,
or the phannaceutically acceptable salts thereof.
Further provided is a compound of forrnula 11 wherein
R2 is -CN, -SCN, and the E or Z isomers of -C(=NORlo)NH2, -C(=NRlo)H,
-C(= NNRlo)NH2, -C(= NRlo)NH2, (:(= NORIo)H, and -C(= NOH)N(Rlo)H;
Rlo is hydrogen, Cl-C4 alkyl, -Ac, ~(O)OCH3, -C(O)OC2H5, -phenyl, -PO2-O-
cation+, ~CO-CH(AA)NH2, -CO-C6H6-NRI IR12, and -CO-C6H6-CH2-NRI ~R12;
AA is-CH3, ~H(CH3)2, -CH2CH(CH3)2,-CH2OH, -CH(OH)CH3, -CH2CO2H,
-CH2CH2CO2H, -(CH2)4NH2. and -(CH2)3-NH2;
R1~ and R12 may be the sarne or diffe-enl and are hydrogen and Cl-C4 aJkyl;
Rll and R12 taken together are morpholinyl, piperazinyl, pyrrolidinyl, piperadinyl, and
N-(CI-C4 alkyl)piperazinyl;
A is CH, CH2, and N;
X is C-R4, and NH;
R4 is -CH3 ~CH3, and -OAc;
R3, preseDt when the connection between positions 2 and 3 is a single bond, is
hydrogen;
R5, R6, R7, R8 may be the sa ne or different and are hydrogen and fluorine;
W is oxygen;
Yl is-CI, -F, -Br, -CH3, -CF3, -CHO, -C02CH3,-OH, -OCH3 ~Ac, -CN, -NO2,
-SH, and-SCH3;
Y2 is hydrogen;
Yl and Y2 taken together are -CH=CRI9-CH=CH-;
Rl9 is hydrogen, -Cl, -Br, -NO2, -CF3, -CO2CH3, -OH, -CN, and -OAc;
provided, however, when Yl is -OCH3, at least one of Rs, R6, R7 or R8 is fluorine,
when Yl is -CF3, the connection between positions 3 and ~ is a double bond,
when A is N, X is C-R4 and R4 is -OCH3 or -OAc and the connection between position
- SUBSTITUTE SHEET
.. . . .... .... . . . . .
.. . ..
- . ... - .. ` . . : ~ -
. . : -
.
: ~ :. . .. ..
-`: ~ : ~ . . ~ . . ..
WO 92/16508 21 D 1 ~ 3 ~ PCr/US92/01746
3 and 4 is a double bond,
when R4 is -CH3, R3 is hydrogen,
when R4 is -CH3, Yl is-CI, -F, -Br, -CF3, -CN, or -NO2, and
when R5 is fluorine, R6, R7 and R8 are each hydrogen;
S or the pharrnaceutically acceptable salts thereof.
llle invention further provides compourds of formula I wherein
~2 is -CN, -N3, -SCN, and the E or Z isomers of -C(=NORlo)NH2, -C(=NRlo)H,
-C~=NNRlo)NH2, -C(=NRIo)NH2, -C(=NORlo)H, and -C(=NOH)NtRlo)H;
Rlo is hydrogen, Cl-C4 alkyl, -Ac, -C(O)OCH3, -C(O)OC2H5, -phenyl, -PO2-O-
cation+, -CO-CH(AA)NH2, -CO-C6H6-NR~ ~R~2, and -CO-C6H6-CH2-NRI ~R~2;
AA is-CH3, -CH(CH3)2, -CH2CH(CH3)2,-CH2OH, ~H(OH)CH3, -CH2CO2H,
-CH2CH2C02H. -(CH2)4NH2. and-(CH2)3-NH2;
Rl I and R12 may be the sarne or different and are hydrogen and Cl-C4 alkyl;
Rll and R12 talceD together are morpholinyl, piperazinyl, pyrrolidinyl, piperadinyl, and
N-(CI-C4 allcyl)piperazinyl;
R3, present when the connection between positions 2 and 3 is a single bond, is
hydrogen, Cl-C4 allcyl, -(CH2),CN, and -(CH2)bR~3;
R~3 is-N(CH3)2, -OH, -OPO3H, -OCH2CH(OH)CH2OH, -OCH2CO2H,
-O2C(CH2)2CO2H, 4R14, and-NRl4;
R14 is a N-termina~ arnino acid;
a is zero to six;
b is one to five;
A is CH, CH2, and CHNtCH3)2;
X is CH, CH2, and C-R4;
R4 is -CH3, -OCH3, and -OAc;
R5, R6, R7, and R8 may be the same or different and are hydrogen and fluorine;
W is oxygen and sulfur;
Z is a linker selected from the group consisting of -(CO)-, -CH=CH-CH=CH- (all E),
-CR15=CRl6- (E or Z), or -CH=C=CH-;
R15 and R16 may be the same or different and are hydrogen and fluorine;
Yl is hydrogen, -I~l, -F, -Br, -CH3, -CF3, -~CH2)CR17, -CHO, -C02CH3, -OH,
-OCH3 -OAc, -CN, -NO2, -SH, and -SCH3;
c is zero to five;
R17 is-CH2N(CH3)2, -OH, -OPO3H, -CH2N(C2Hs)2, -O-sugar,
-OCH2CH(OH)CH2OH, -OCH2CO2H, -O2CCH2CH2CO2H, -CH2N= CHN(CH3)2, -OR18,
and -NR18;
- SUBSTITUl'E SHEET
. . .
. . , , . . ~ :
- . .,
:: -
WO 92/16508 2 ~ 013 3 S PCI/-rS92/01746
R18 is a N-terminal arnino acid;
Y2 is hydrogen, -Cl. -F, -OCH3, and -CF3;
Yl and Y2 taken together are -OCH20-, -OC(CH3);!0-, -OCH2NH-, -NHCH2~,
-OCH2S, or-SCH20-;
S provided, however, when Z is -CR15=CR16- (E), and Y2, R15, and R16 are each
hydrogen, Yl is not hydrogen,
when R4 is -CH3, R3 is hydrogen,
when R4 is -CH3, Yl is -Cl, -F, -Br, -CF3, -CN, or -N02, and
wben R5 is fluorine, R6, R7 and R8 are each hydrogen;
or the pharrnaceutically acceptable salts thereof.
The invention aJso provides a method for treating a human infected wi~ one or more
than one strain of a human immunodeficiency virus (HIV) which comprises administering an
effective amount of a compound of formula I wherein
R2 is -CN, -N3, -SCN, and the E or Z isomers of -C(=NORlo)NH2, -C(=NRlo)H,
-C(=NNR1o)NH2, -C(=NRlo)NH2, -C(=NORIo)H, and -C(=NOH)N(Rlo)H;
Rlo is hydrogen, Cl-C4 alkyl, -Ac, -C02CH3, -C02C2H5, -phenyl, -P02-0- cation+,
-CO-CH(AA)NH2, -CO-C6H6-NRI ~R12, and -CO-C6H6-CH2-NR1 IR12;
M is-CH3, -CH(CH3)2, -CH2CH(CH3)2,-CH20H, -CH(OH)CH3, -CH2C02H,
-CH2CH2C02H, -(CH2)4NH2. and-(CH2)3-NH2;
R11 and R12 may be the sarne or different and are hydrogen and Cl-C4 alkyl;
Rll and R12 tal~en together are morpholinyl, piperazinyJ, pyrrolidinyl, piperadinyl, and
N-(CI-C4 allcyl)pipera~inyl;
R3, present when the connection between positions 2 and 3 is a single bond, is
hydrogen, Cl-C4 allcyl, -(CH2)"CN, and -(CH2)bR~3;
R13 is-N(CH3)2, -OH, -OP03H, -OCH2CH(-OH)CH20H, -OCH2C02H,
-02C(cH2)2co2H~ -ORI4, and -NRI4;
R14 is a N-terminal arnino acid;
a is zero ~o s~x;
b is one to five;
A is CH, CH2, N, and CHN(CH3)2;
X is CH, CH2, NH, O, and C-R4;
R4 is -CH3 ~CH3, and -OAc;
R5, R6, R7, and R8 may be the same or different and are hydrogen and fluorine;
W is oxygen and sulfur;
Z is a direct ~ond or a linker selected from the group consisting of -(CO)-,
-CH=CH-CH=CH- (all E), -CR15=CRl6- (E or Z), and -CH=C=CH-;
SUBSTITUTE SHEET
- . ' . . ' ,
. ' ' . . .
., . - . , . `, ` . .. .
. . . -
. . . . :.
. . .
-. . , - - , , .
. . . .. . . . .. . ..
~vo 92/16508 2 1 ~ ~ 3 3 ~ -8- PCI/US92/01746
R15 and R16 may be Lhe same or different and are hydrogen and fluorine;
Yl is hydrogen, -Cl, -F, -Br, -CH3, -CF3, -(CH2)cRl7, ~HO, -C02CH3. -OH,
-OCH3 -OAc, ~N, -N02, -SH, and -SCH3;
c is zero ~o five;
R17 is ~H2N(CH3)2, -OH, -OP03H, -CH2N(C2Hs)2, -O-sugar~
-OCH2CH(OH)CH20H, -OCH2CO2H, -02CCH2CH2C02H, -CH2N=CHN(CH3)2, -OR~8, or
-NR18;
R18 is an N-terminal amino acid;
Y2 is hydrogen, -Cl, -F, -OCH3, and -CF3;
Yl and Y2 taken together can forrn -OCH20-, -OC(CH3)20-, -OCH2NH-,
-NHCH20-, ~CH2S-, -SCH20-, and-CH=CRIg-CH=CH-;
R19 is hydrogen, -Cl, -Br, -N02, ~F3, -C02CH3, -OH, -CN, and -OAc;
provided, however, when Z is a direct bond and R2 is ~(=NOH)N(RIo)H and Rlo is
hydrogen, Rl9 is not hydrogen,
when Z is a direct bond, Yl is not hydrogen,
when Yl is -OCH3 and Z is a direct bond, at least one of Rs, R6, R7 or R8 is fluorine,
when Y1 is -CF3 and Z is direct bond, the coMection between positions 3 and 4 is a
double bond,
when Z is a direct bond, X is not 0,
when Z is a direct bond, R2 is not -N3,
when Z is a direct bond, Y2 is hydrogen or -F,
when A is N, X is C-R4 and R4 is -OCH3 or -OAc and the connection between position
3 and 4 is a double bond,
when A is CHN(cH3)2~ X is CH
when R4 is -CH3, R3 is hydrogen,
when R4 is -CH3, Yl is -Cl, -F, -Br, -CF3, -CN, or -N02,
when R5 is fluorine, R6, R7 and R8 are each hydrogen,
when Yl and Y2 taken together are -CH=CRI9-CH=CH-, Z is a direct bond, and
when Yl and Y2 taken together can forrn -OCH20-, -OC(CH3)2~, -OCH2NH-,
30 -NHCH20-, -OCH2S-, and -SCH20-, Z is a linker; to a human patient.
The preferred compound for this method is l-E-(3-(4-chlorophenyl)-1-o~o-2-propenyl)-
2-cyano-1 ,2-dihydroquinoline.
The invention further provides the use of the compounds of the invention and/or their
pharmaceutically acceptable salts, hydrates, and solvates for the preparation of pharmaceutical
35 formulations. These formulations are useful to practice the method claimed in this invention.
DETAILED DESCRIPllON OF THE INVENTION
SUBSTITVTE SHEET
- - - - . , ` . - - .
.~: . . :
-. ` '
- ~
wo 92/16508 21 a 13 3 ~ . PCl/US92/01746
r 9
Generally, the compounds of forrnula I are preferred.
Particularly preferred are the compounds of the invention wherein there is a double
bond berween A and X, and A is CH. It is preferred that X is CH. It is preferred that Z is
~R15=CR16- (E), and that R15 and R16 are hydrogen. It is preferred that W is O. It is
5 preferred that R3 and R5 are hydrogen and R6 R7 and R8 are eithe- hydrogen or F. It is
preferred tha~ R2 is CN. It is preferred that Y2 is hydrogen. It is preferred that Y I is Cl or
C:N, most preferred is where Y I is Cl.
It is understood by those sl~illed in the art that the stereoisomers and enantiomers of the
compounds of the invention are included within the scope of the inventioD.
The compounds of the invention are useful as inhibitors of viral reverse transcriptase,
an enzyme necessary for human immunodeficiency virus replication, and tnerefore would be
useful in the treatment of disease, such as acquired immune deficiency syndrome (AIDS).
The term human retrovirus ~HRV) includes human immunodeficiency virus type I
(HIV-I), or variant strains thereof, apparent to one skilled in the art, which belong to the same
15 viral families and which create similar physiological effects in humans as HRV.
Patients to be treated would be those individuals: I) infected with one or more than one
strain of a buman retrovirus as determined by the presence of either measurable viral antibody
or antigen in the serum and 2) having either a symptomatic AlDS defining infection such as (a)
dissèminated histoplasmosis, (b) isopsoriasis, (c) bronchial and puln onary candidiasis including
20 pneumocystic pneumonia, (d) non-Hodgkin's Iymphoma, or (e) Kaposi's sarcoma and being less
than si~ty years old; or having an absolute CD4 Iymphocyte count of less than 500/mm3 in the
peripheral blood. Additionally, persons who show the signs and symptoms of AI~S-re1ated
comple~ (ARC) may be treated with the compounds of this invention Sucb signs andsymptoms include, but are not limited to, generali~ed Iymph adenopathy, weight loss, anemia,
25 candidiasis, and imrnunologic abnormalities characteristic of AIDS. Treatment of ARC and
AIDS patients would consist of maintaining an inhibitory level of the compound of the
invention used according to this invention in the patient at all times and would continue until
the occurrence of a second symptomatic AIDS defining infection indicates alterrlate therapy is
needed. Treatment of asymptomatic patients would typically require lower daily maintenance
30 dosages. In addition, the compounds of the invention may be used in conjunction with other
antiviral agents such as AZT.
The compounds of the invention, or pharmaceutically acceptable salts, hydrates, and
solvates thereof, can be used and administered in practicing the method claimed in this
invention. The compounds of the invention may form acid addition salts when reacted with
35 acids of sufficient strength to produce the corresponding salt. ln addition, some of the variable
substituents are acids and thus form base addition salts when reacted with bases of sufficient
SUBSTITUTE SHEET
. . . .. . . . . . . .
:.' : . . - . , .
- , . . - , . ~ -
- .,
-
WO92/1650X 2~ PCT/I~'S92/01746
1~ ~
strength. Pharmaceutically acceptable salts refers to those salts of the compounds of the
invention which would be readily apparent to a manufacturing pharmaceutical chemist to be
equiva]ent to, or bener than, the parent compound in properties such as formulation, stability,
patient acceptance, and bioavailability. Generally, the pharmaceutically acceptable salts, either
5 acid or base as the case may be, are preferable over the free acid or base since the salt is more
water soluble and more crystalline. The pharmaceutically acceptable salts include botb
inorganic and organic acids and bases. Examples of pharmaceutically acceptable salts include
salts of the following acids: methanesulfonic, hydrochloric, hydrobromic, sulfuric, pbosphoric,
niuic, benzoic, tartaric, fumaric, maleic, p-toluenesulfonic, benzenesulfonic, and the lilce.
10 E~amples of pharmaceutically acceptable salts include salts of the following bases: hydroxide,
ammonia, tromethamine (l~AM), and the like. Suitable cations include, for e%ample. sodium,
potassium, calcium, and magnesium.
Those slcilled in the art would know how to formulate the compounds used to practice
the method claimed in this invention into appropriate pharmaceutical dosage forms. E~amples
15 of the dosage forms include oral formulations, such as tablets or capsules, or parenteral
formulations, such as sterile solutions.
When the compounds used to practice the method claimed in this invention are
administered orally, an effective amount is from about 0,2 to about 100 mg per Icg per day for
asymptomatic patients, and about I to about 500 mg per kg per day for ARC and AIDS
20 patients. A typical unit dose for a 70 kg human AlDS patients would be from about 50 mg to
1000 mg, preferably 200 mg to 1000 mg taken one to four times per day. Either solid or fluid
dosage forms can be prepared for oral administration. Solid compositions are prepared by
r~u%ing the compounds used to practice the method claimed in this invention with conventional
iDgredients such as talc, magnesium stearate, dicalcium phosphate, magnesium aluminum sili-
2S cate, calcium sulfate, starch, lactose, acacia, methyl cellulose, or functionally similarpbarmaceutical diluents and carriers. Capsules are prepared by mi%ing the compounds used to
practice the method claimed in this invention with an inert pharmaceutical diluent and placing
the mixture into aD appropriately sized hard gelatin capsule. Soft gelatin capsules are prepared
by machine encapsulation of a slurry of the compounds used to practice the method claimed in
30 this invention with an acceptable inert oil such as vegetable oil or light liquid petrolatum.
Syrups are prepared by dissolving the compounds used to practice the method claimed in this
invention in an aqueous vehicle and adding sugar, aromatic flavoring agents and preservatives.
Eli%irs are prepared using a hydroalcoholic vehicle such as ethanol, suitable sweeteners such as
sugar or saccharin and an aromatic flavoring agent. Suspensions are prepared with an aqueous
35 or organic vehicle and a suspending agent such as acacia, tragaca~th, or methyl cellulose.
When the compounds used to practice the method claimed in this invention are
SUBSTITUTE SHEET
~ .
~:
WO 92/16508 2 ~ O ~ 3 ~ 5 PCT/US92/0174b
administered parenterally, it can be given by injection or by intravenous infusion. An effective
amount is from about I to 100 mg per kg per day. Parenteral solutions are prepared by
dissolving the compounds used to practice the method claimed in this invention in an
appropriate solvent and filter sterilizing the solulion before placing in a suitable sealable vial or
5 ampule. Parenteral suspensions are prepared in substantially the same way except a sterile
suspension vehicle is used and the compounds used to practice the method claimed in this
invention are sterilized with ethylene oxide, suitable gas or other methods Lnown in the art,
before it is suspended in the vehicle.
The exact route of administration, dose, or frequency of administration would be10 readily determined by one skilled in the art and is dependent on the particular compound of the
invention being used, and on the age, weight, general physical condition, other medication the
individual is taking, or other clinical symptoms specific to the patient to be treated.
Tenns used in this specification and claims have the following definitions:
~AIDS~ means acquired immune deficiency syndrome.
~ARC" means All)S-related complex.
"E or Z" designates stereochemistry around a double bond. The two groups attached to
each atom of the double bond are assigned priorities according to the Prelog-Cahn-lngold
system. "E~ refers to an arrangement in whicb the two higber priority or two lower priority
groups are on the opposite sides of the double bond. ~Z" refers to an arrangement in which the
20 two higher priority groups are on the same side of the double bond
"Phenyl~ mèans -C6H5
"N-terminal arnino acid~ means a naturally occurring amino acid, and synthetic
derivatives thereof, in which the arnino group is free, i.e., the amino acid is linked via the
carboxyi group to form either an ester or an arnide.
"-Ac~ means-C(O)CH3; ~-OAc~ means-OC(O)CH3.
~O-sugar~ means a hexose or pentose monosaccharide linked at the 2-hydroxyl group.
~-C02-~ or "-C(O~O-~ designates an es~er linked via the carbon.
~-O2C-~ or ~-OC(O)-~ also designates an ester, with the linkage Yia the oxygen.
~-CO2H~ designates a carboxylic acid.
"-CH=CRIg-CH=CH-~ designates a carbon chain linking Yl and Y2 where (Yl end)
-CH-CR19-CH=CH- (Y2 end).
Throughout the disclosure and clairns, common shorthand chemical terms are used. The
carbon atom content of the various hydrocarbon-contain. 3g moieties is indicated by designating
the minimum and maxirmurn number of carbon atoms in the moiety, i.e. Cm-Cn indicates a
moiety of integer ~m~ to the integer ~n~ carbon atoms, inclusive and includes the isomeric
forms. For exarnple, Cl-C6 alkyl refers to an alkyl of one to six carbons, inclusive, including
SUBSTITUTE SHEET
.
`
wo 92/16508 ~ :1 0 1 3 3 ~ PCr/US92/01746
-12-
the isomeric forms. Chemical formulas, or portions thereof, drawn in a linear fashion may use
Ihe symbol ~ o represent a single chemical bond between atoms in the linear chain. Likewise,
the symbol ~ = ~ represents a double bond, and ~ . ~ a uiple bond, between atoms in the chain.
In ~he structural formulas the symbol ~ represents a bond between two atoms which may
be a single or a double bond; the symbol ~ represents a bond which may or may not be
present, and if present, is a single bond.
Temperatures are in degrees Celsius. The letter "h~ means hours. "TLC~ means thin
layer chromatography. MS refers to mass specuometry. MS data are expressed as m/e or
mass/change unit.
Svnthesis of Representative Compounds
In preparative methods that follow, the compounds of the invention are synthesized
following techniques which are known, or readily acquired, by one skilled in the art. E~amples
of such techniques are:
F.D. Popp, ~Reissen Compounds and Related N-acyldihydroquinolines~ in Quinolines,
Par~ Il, Ed. G. lones, (1982) p. 353-372, is a general review covering the synthesis and
reactions of Reissen compounds. This review specifically describes the synthesis of 1-(4-
chlorobenzoyl)-2-cyano- 1,2-dihydroquinoline, 1 -(4-fluorobenzoyl~2-cyano- 1,2-
dihydroquinoline, 1-(4-me~hoxybenzoyl)-2-cyano-1,2~ih~droquinoline, 1-(4-toluoyl)-2-cyano-
1,2-dihydroquinolhe, 1-(~nitroben~oyl)-2-cyano-1,2-dihydroquinoline, 1-(2-naphthoyl~2-
cyano-1,2-dihydroquinoline, and 1-(1-o~o-3-phenyl-2-propenyl)-2-cyano-1,2-dihydroquinoline.
F.D, Popp, W. Blount and P. Melvin, 1. Q~. Chem., 26, 4930 (1961), describes themethylene chloride/water method for synthesizing Reissert Compounds.
S. Ruchirawat, N. Phadungkul, M. Chuankamnerdkarn, and C. Thebtaranonth,
Heterocvcles, 6, No. 1, 43 (1977), describes the trimethylsilyl cyanide method of synthesizing
Reissert compounds.
M. Rozwadowska, Roczniki Chemii. 51, 2321 (1977), describes hydrogenation of
dihydroquinoline Reisser~ compounds to tetrahydroquinolines.
L. Walters, M. Siegel, and R. Cook, L. Heterocvclic Chem., 5, 577 (1968), describes
synthesis of amido%imes, also called carboxamidamides, via addition of hydro%ylamine to a
Reissert compound.
H. Bartsch, O. Schwarz, and G. Neubauer, Heterocycles, 24, 3483 (1986), describes
the synthesis of a benzoxazine Reissert compound.
F. Elroy and R. Lenaers, "The Chemistry of Amido%imes and Related Compounds,~
Chem. a~ev. 155 (1962), describes synthesis and reactions of amido~imes.
Uff, B.C., Budhram, R.S., Consterdine7 M.F., Hicks, J.K., and Slingsby, 33.P.,
1 ~h~. Soç., Perkins7 Trans a (18):2018-22 (1977), describes synthesis of 1-(4-
SUBSTITUTE SHEET
:, . . . . ~ ~
- . . . .
. . : ~ . - .. ~
wo 92/16508 2 ~ O 1 3 3 ~ PCI /US92/01746
f
-13-
methoxybenzoyi)-2-cyanoJ,-me~hyl- I ,2-dihydroquinoline.
F.D. Popp and A. Soto, I Chem. Soc., 1760 (1963), describes the synthesis of 1-(~
fluorobenzoyl)-2-cyano- 1 ,2-dihydroquinoline.
All starting materials are icnown and are corrunercially available, or are readily
prepared from icnown or readily available materials.
The synthesis of compounds of the invention proceed, generally, as outlined below and,
more specifically, in the representative examples that follow. The synthesis of the dihydro- and
teuahydroquinolines, the dihydroquinazolines and the dihydroquinoxalines are outlined in Chart
A, the teuahydrobenzoxazines in Chart B, and ti~e a nidoxime compounds of the invention in
Chart C. In the Chans the l-(benzoyl) or l-(cinnamoyl) substituents are designated "R~, and
the halide of the acid halide is designated ~X~.
The exarnples that follow are illustrative and not limitations of the preceding disclosure.
The conditions described here and in the examples for the synthesis and purification of the
compounds of the invention may be altered depending on the choice of reactants and solvents,
the batch size, the degree to which the reaction is to be carried to completion, and other factors
of concern to the chemist.
Procedure for ~reoaration of Reissert compounds (Chart A~.
There are numerous techniques available for the synthesis of Reissert compounds from
quinoline. Among these are preparation in aqueous potassium cyanide, potassium cyanide in a
dimethylformamide-water mixture, liquid sulphur dioxide as a solvent, anhydrous benzene-
liquid hydrogen cyanide, and trimethylsilyl cyanide-aluminum chloride in methylene chloride.
See, F.D. Popp, ~Reissert Compounds and Related N-acyldihydroquinolines~ in Quinolines,
Part II, Ed. G. Jones, (1982) p. 353-372. We prefer the methylene chloride-trimethylsilyl
cyanide system.
Referring now to Chart A, one equivalent (I eq.) of the quinoline (Chart A fig. 1),
quinazoline (Chart A, fig. 4), quinoxaline (Chart A, fig. 5), or substituted forms thereof, is
dissolved in dichloromethane or acetonitrile to a concentration of 0.4-0.5 M. An acid halide,
preferably chloride, (1.5-2 eq.), Lewis acid, preferably aluminum trichloride (anhydrous, 0.05
eq.) and trimethylsilyl cyanide (1.5-2 eq.) are added while stirring. A thio-acid halide is
substituted when the corresponding thio-Reissert compound is desired. Thio-acid halides are
synthesized following known procedures, for example, R. Mayer and S. Scheithauer, J.
Heterocyclic Chem. 16, 1589 (1977). The Reissert reaction is carried out under ambient
temperature conditions, e.g. 23-27 degrees, or refluxing acetonitrile for 8-substituted
compounds, for a time sufficient to effect as complete a reaction as possible, e.g. from 0.5 to
48 hours. The products of the reaction (Chart A, fig. 2) are followed by ll C using solvent
systems known in the art. Upon completion, the reaction is concentrated in vacJlo and then
SUBSTITUTE SHEE~
:
'
: ;
.
Wo 92/16~08 ;~ 3 3 ~ PCr/l,'S92/0174
-14-
placed on a silica gel column and eluted under flash chroma~ography conditions. The product is
recrystallized from a low boiling point aliphatic alcohol, preferably 95% ethanol, or
ethylacetate/hexane.
When a tetrahydroquinoline product is desired, the dihydroquinoline (Chart A, fig. 2) is
5 dissolved in alcohol, preferably 95~o ethanol, and reduced by hydrogenation in the presence of
platinum. The reaction is followed and the product lChart A, fig. 3) recrystallized as outlined
above.
Preparation 1 6,7-difluoroquinoline
Sodium 3-nitrobenzensulphonate ~27.25 B, 1.25 eq.), followed by glycerol (25.50 g,
10 20.22 ml, 2.86 equiv.), and aqueous sulphuric acid (80% w/w, 81 ml) are placed in a 1 L.
three-necked flask. With stirring, 3,4-difluoroaniline (12.50 g, 9.60 rnl, 1 eq.) is slowly added
and then heated under gentle reflux in an oil bath at 175C for 5 hours. The reaction mixture
is cooled to room temperature and treated with aqueous sodium nitrite (25% w/w, 55 ml) from
a dropping funnel. The mi~ture is subjected to steam-distillation. The yellowish aqueous
15 distillate is discarded. The reaction product is cooled in an ice bath, made strongly alkaline,
i.e. pH 9-12, by the cautious addition of aqueous sodium hydroxide (50% w/w), and again
subjected to steam distillation. A colorless solid formed in both the condenser and receiver.
Extraction with ethyl acetate, drying over sodium sulphate, and removal of the solvent in the
rotary evaporator at 35~C, under house vacuum gave a solid (13.20 g) TLC in hexane-methanol
20 (1:6 v/v) showed the presence of a major and a minor product Direct crystallization gave 6,7-
difluoroquinoline (9.25 g) m.p. 112-113C. The mother liquors contain the minor 5,6-
difluoro-isomer.
PreparatiQn 2 6,8-difluoroquinoline
6,8-difluoroquinoline is synthesized by substituting 2,~difluoroaniline for 3,~
25 difluoroaniline in Preparation 1. The remaining procedure and purification are substantially
unchanged.
Preparation 3 7,8 difluoroquinoline
7,8-difluoroquinoline is synthesized by substituting 2,3-difluoroaniline for the 3,4-
difluoroaniline in Preparation 1. The remainder of the procedure and purification are
30 substantially unchanged.
Preparation 4 4-cyanociMamic acid
4-cyanobenzaldehyde (I eq.), malonic acid (I eq.), pyridine (0.8 M), and piperidine (1
ml/10 m] pyridine) are heated at reflux for 24 h. or until carbon dioxide is no longer evolved.
The mixture is evaporated to dryness in vacuo. The residue is redissolved in water and the pH
35 adjusted to 10 using concentrated ammonium hydroxide. The mixture is filtered through celite
and the filter rinse~ with water. The pH of the filtrate is adjus~ed to 2 using IN hydrochloric
SUBSTITUTE SHEET
, .. .. . .. .. - . . .
:: -' - . ' '
....
- .-: - .. : . -
wo 92/16508 2 1 0 1 3 3 ~ PCr/US~2/01746
~- 15-
acid. Chilling precipita~es solids which are collected, dissolved in acetone and 95X ethanol,
and evaporated in vacuo until solids form. Chilling produces crystalline compound (1
cyanocinnamic acid).
Preparation 5 4-cyanocinnamoyl chloride
4{~yanocinnamic acid (Preparation 4, 1 eq.) is dissolved in methylene chloride at a
concentration of 0.5 M. Oxalyl chloride (I eq.) is added and the mixture chilled to 0C. DMF
(0.5 eq.) is added, the mixture stirred at 0C for 0.5 h. warmed to ambient temperature, and
stirred an additional 2 hours. Evaporation in v~cuo yields 4-cyanocinnamoyl chloride.
Example la 1-(4-bromobenzoyl)-2-cyano-1,2-dihydroquinoline
Quinoline (0.104 g, 0.8 mmol) is dissolved in 0.5 ml of methylene chloride. 88 ~L of
trimethylsilylcyanide is added at ambient temperature. 4-Bromobenzoyl chloride (0.25 g.) is
dissolved in 0.5 mL of methylene chloride is added following 5 mg of aluminum trichloride.
After 40 min. the reaction is filtered through a pad of silica gel and the pad is washed with 20
mL of methylene chloride. The combined filtrates are washed first with 10 mL of water, then
15 with 10 mL of saturated aqueous sodium bicarbonate and finally S mL of water. The organic
layers are concentrated in ~Jac~lo to afford an oil which is dissolved in 4 ml of 9S percent
ethanol. This is evaporated to 2.5 mL and cooled at 0C for 30 min. The crystals are
collected and dried in a vacuum oven to afford 85 mg of 1-(4-bromobenzoyl)-2-cyano-1,2-
diSydroquinoline (m.p, 149-151C). The mother liqùor is evaporated and triturated with I rnL
20 of 9S percent ethanol. The crystaJs formed are collec~ed and dried to afford 31 mg of a second
crop.
In an analogous manner, the following compounds are synthesized using the designated
acid chloride.
E%asnple Ib 1-(4-fluorobenzoyl)-2-cyano-1,2-dihydroquinoline
Following the procedure of Exarnple la, 4-fluorobenzoyl chloride is used as the acid
chloride and the product, 1-(4-fluorobenzoyl)-2-cyano-1,2-dihydroquinoline, has a m.p. 114-
116C.
Exarnple 2 1-(4-trifluoromethylbenzoyl)-2~yano-1,2-dihydroquinoline
Following the procedure of Exarnple la, 4-trifluoromethylbenzoyl chloride is used as
the acid chloride. The product, 1-(4-trifluoromethylbenzoyl)-2-cyano-1,2-dihydroquinoline, has
a m.p. 135-136C.
Example 3 1-E-(I-oxo-3-phenyl-2-propeDyl)-2-cyano-1,2-dihydroquinoline
- Following the procedure of E~arnple la, I-E-ci ~namoyl chloride is used as the acid
chloride. The product. l-E-(I-oxo-3-phenyl-2-propenyl)-2-cyano-I,2~ihydroquinoline, has m.p.
149-150~.
Exarn~le 4 1 -E-(3-(~trifluoromethylphenyl)- 1 -oxo-2-propenyl)-2-cyano- 1,2-
SUBSTITUTE SI~IEET
.
~VO 92f16508 2 ~ 3 3 ~ PC~/US92/0174b
dihydroquinoline
Following the procedure of Exarnple la, E-4-trifluoromethylcinnarnoyl chloride is used
as the acid chloride. The product, I-E-(3-(4-trifluoromethylphenyl)-1-oxo-2-propenyl)-2-cyano-
1,2-dihydroquinoline, has a m.p. 172-173C.
Exarn~le 5 1-(4-carbomethoxybenzoyl)-2-cyano-1,2-dihydroquinoline
Following the procedure of Exarnple 1, 4-carbomethoxybenzoyl chloride is used as the
acid chloride. The product, 1-(4-carbomethoxybenzoyl)-2-cyano-1,2-dihydroquinoline, has a
m.p. 165-166C.
Exarngle 6 1 -(4-cyanobenzoyl)-2-cyano- 1,2-dihydroquinoline
Following the procedure of Example la, 4-cyanobenzoyl chloride is used as the acid
chloride. The product, 1-(4-cyanobenzoyl)-2-cyano-1,2-dihydroquinoline. has a m.p. 154-
155C.
Exarngle 7 1-E-(3-(4-chlorophenyl)-1-oxo-2-propenyl)-2-cyano-1,2-dihydroquinoline
Following the procedure of Example la, E4-chlorocinnarnoyl chloride is used as the
acid chloride. The product, I-E-(3-(4-chlorophenyl)-1-oxo-2-propenyl)-2-cyano-1,2-
dihydroquinoline has a m.p. 166-167C.
Exarn~le 8 1 -E-(3-(4-fluorophenyl)- 1 -oxo-2-propenyl)-2-cyano- 1,2-dihydroquinoline
Following the procedure of Exa nple la, E4-fluorocinnamoyl chloride is used as the
acid chloride. The product, l-E-(3-(~fluorophenyl)-1-oxo-2-prophenyl)-2-cyano-1,2-
dihydroquinoline has a m.p. 142- 143C.
Example 9 1-E-(2-fluoro-1-oxo-3-phenyl-2-propenyl)-2-cyano-1,2-dihydroquinoline
Fol~owing the procedure of Example la, E-~x-fluorocinnamoyl chloride is used as the
acid chloride. The product, I-E-(2-fluoro-1-oxo-3-phenyl-2-propenyl)-2-cyano-1,2-
dihydroquinoline, has a m.p. 1~2-153C.
Example 10 1-E-(3-(3,4-dichlorophenyl)-1-oxo-2-propenyl)-2-cyano-1,2-dihydroquinoline
Following the procedure of Example la, E-3,4-dichlorocinnamoyl chloride is used as
the acid chloride. The product, I-E-(3,4-dichlorophenyl-1-oxo-2-propenyl)-2-cyano-1,2-
dihydroquinoline, has a m.p. 152-153C.
Example 11 1 -E-(3-(4-cyanophenyl)- 1 -oxo-2-propenyl)-2-cyano- 1,2-dihydroguinoline
Following the procedure of Example la, E4-cyanocinnamoyl chloride (Preparation 5)
is used as the acid chloride. The product, I-E-(3-(4-cyanophenyl)-1-oxo-2-propenyl)-2~yano-
1,2-dihydroquinoline has a rn.p. 176C.
Ex n~le 12 1-E-(3-(3,4-methylenedioxyphenyl)-l~xo-2-propenyl)-2-cyano-1,2-
dihydroquinol ine
Following the procedure of Example la, E-3,4-methylenedioxyciMarnoyl chloride isused as the acid chloride. The producl, I-E-(3-(3,4-methylenedioxyphenyl)-l~xo-2-propenyl)-2-
SUBSTiTUTE SHEE~
.. . . :
. ' ` '
-
.~ . .
wo 92/1~508 2 1 0 1 ~ PCI/l~S92~01~46
r . . . 1 7 , .
cyano-1,2~ihydroquinoline, has a m.p. 176-177C.
Exam~le 13 1 -E-(3-(4-methoxyphenyl)- 1 -oxo-2-propenyl)-2-cyano-1,2~ihydroquinoline
Following the procedure of Exarnple la, E-4-methoxycinnamoyl chloride is used as the
acid chloride. The product is l-E-(3-(4-methoxyphenyl)-1-oxo-2-propenyl)-2-cyano-1,2-
5 dihydroquinoline.
Calcd.: C,75.93; H, 5.10; N, 8.85
Found: C, 75.74; H, 5.31; N, 8.86E~,arnple 14a 1-E-(3-(4-acetoxyphenyl)-1-oxo-2-propenyl)-2-cyano-1,2-dihydroquinoline
Following the procedure of Exarnple la, E4-acetoxycinnamoyl chloride is used as the
10 acid chloride. The product is l-E-(3-(4-acetoxyphenyl)-1-oxo-2-propenyl)-2-cyano-1,2-
dihydroquinol ine.
Calcd.: C, 73.24; H, 4.68; N, 8.13
Found: C, 72.94; H, 4.51; N, 7.99
Exarnple 14b 1 -E-(3-(4-carbomethoxybenzoyl)- I -oxo-2-propenyl)-2-cyano- 1,2-
lS dihydroquirJoline
Following ~he procedure of Exarnple la, E4-carbomethoxycinnarnoyl chloride is used
as the acid chloride. The product, I-E-(3~4~arbomethoxybenzoyl)-1-oxo-2-propenyl)-2-cyano-
1,2~ihydroquinoline, has a m.p. 168-170~C.
~am~ l-E-(3-(4-methylphenyl)-1-oxo-2-trans-propenyl)-2~yano-1,2-dihydroquinolineFollowing the procedure of E%ample la, E-(4-methylcinnamoyl chloride) is used as the
acid chloride. The product is l-E-(3-(4-methylphenyl)-1-oxo-2-propenyl)-2-cyano-1,2-
dihydroquinoline.
Calcd.: C, 79.98; H, 5.37; N, 9.33
Found: C, 79.54; H, 5.40; N, 9.17
25 Example 16 1-(2-napthoyl)-2-cyano-1,2~ihydroquinoline
Following the procedure of Example la, 2-napthoylchloride is used as the acid chloride.
The product, 1-(2-napthoyl)-2-cyano-1,2-dihydroquinoline, has a m.p. 185-186C.
Exasnple 17 1 -(4-chlorobenzoyl)-2-cyano~methyl- 1,2-dihydroquinoline
Following the procedure of Example la, 4-methylquinoline is substituted for guinoline
30 and 4-chlorobenzoyl chloride is used as the acid chloride. The product, 1-(4-chlorobenzoyl~2-
cyano 1 methyl-1,2-dihydroquinoline, has a m.p. 173-174C.
Calcd. for C18Hl3CIN2O: C, 70.02; H, 4.24; N, 9.08; Cl, 11.48.
Found: C, 69.95; ;H, 4.28; N, 8.90; Cl, 11.45.
Example 18 1-(4-chlorobenzoyl)-2-cyaDo S-fluoro-1,2-dihydroquinoline
Following the procedure of Example la, 5-fluoroquinoline is substituted for quinoline and
4-chlorobenzoyl chloride is used as the acid chloride. The product. 1-~4-chlorobenzoyl)-2-
SUBSTITUTE SHEET
.
wo 92/16~0X PCr/US92/01746
2 1 0 1 ~ ~ 5 --18
cyano-5-fluoro-1,2~ihydroquinoline, has a m.p. 144-145C.
Calcd. for C17HloClFN2O: C, 65.29; H, 3.22; N, 8.96; Cl, 11.34; F, 6.08.
Found: C, 65.37; H, 3.24; N, 9.03; Cl; 11.32; F, 6.47.
E~.arnple 19a 1 -(4-chlorobenzoyl)-2-cyano~fluoro- 1,2-dihydroquinoline
Following the procedure of Exarnple la, 6-fluoroquinoline is substituted for quinoline and
4~hlorobenzoyl chloride is used as the acid chloride. The product, 1-(4-chlorobenzoyl)-2-
cyano~fluoro-1,2-dihydroquinoline, has a m.p. 147-148C.
Calcd. for C17HloClFN20: C, 65.29; H, 3.22; N, 8.96; Cl, 11.34; F, 6.08.
Found: C, 65.18; H, 3.27; N, 8.87; Cl, 11.09; F, 6.36
Exarnple 19b 1-(4-chlorobenzoyl)~fluoro-2-cyano-1,4-dihydroquinoline
The product of Exarnple 19a is subjected to reverse-phase HPLC separation (isocratic 40
THF/60% wa~er mobile phase). Pure 1-(4-chlorobenzoyl)~fluoro-2-cyano-1,4-
dihydroquinoline is isolated (m.p. 14~150C).
Exarnple 20 1-(4-chlorobenzoyl)-2-cyano-7-fluoro-1,2-dihydroquinoline
Following the procedure of Example la, 7-fluoroquinoline is substituted for quinoline and
4-chlorobenzoyl chloride is used as the acid chloride. The product, 1-(4-chlorobenzoyl)-2-
cyano-7-fluoro-1,2~ihydroquinoline, has a m.p. 174-175C.
Calcd, for C17HloClFN2O: C, 65.29; H, 3.22; N, 8.96; Cl, 11.34; F, 6.08.
Found: C, 65.23; H, 3.27, N, 8.98; Cl, 10.99; F, 6.19.
Example 21 1-(4-chlorobenzoyl)-2-cyano-8-fluoro-1,2-dihydroquinoline
Following the procedure of Example la, 8-fluoroquinoline is substituted for quinoline and
4-chlorobenzoyl chloride is used as the acid chloride in acetonitrile solvent and the solution
refluxed as described above. The product, 1-(4-chlorobenzoyl)-2-cyano-8-flùoro-1,2-
dihydroquinoline, has a m.p. 156-157C.
Calcd. for C17HIoClFN2O: C, 65.29; H, 3.22; N, 8.96; Cl, 11.34; F, 6.08.
Found: C, 65.36; H, 3.23; N, 8.92; Cl, 11.16: F, 6.02.
Example 22 1-(4-methoxybenzoyl)-2-cyano-6,7-difluoro-1,2-dihydroquinoline
Following the procedure of Example la, 6,7-difluoroquinoline (Preparation 1) is
substituted for quinoline and 4-methoxybenzoyl chloride is used as the acid chloride. m.p. 139-
140C.
Calcd. for C~8H~2F2N2O2: C, 66.25; H, 3.71; N, 8.59; F, 11.65.
Found: C, 66.65; H, 3.73; N, 8.47; F, 12.09.
Exa nple 23 1-(4-chlorobenzoyl)-2-cyano-1,4-dihydroquinoxaline
Following the procedure of Exarnple la, quinoxaline is used in place of quinoline and 4
chlorobenzoyl chloride is the acid halide. The product, 1~4-chlorobenzoyl)-2-cyano-1,4-
dihydroquinoxaline, has a m.p. 231-232C.
SUBSTITUTE SHEE~
..
.
- . - :: .
WO 92/16~08 2 1 Q 1 3 3 ~ PCT/VS92/01746
_ 1 9_
CaJcd. for C16HloClN3O: C, 64 98; H, 3.40; N, 14.21.
Found: C, 65.05; H, 3.55; N, 14.30.
xarnple 24 1-(4-chlorobenzoyl)-2-cyano~,7~ifluoro-1,2-dihydroquinoline
Following the procedure of Exarnple la, 6,7-difluoroquinoline (Preparaflon 1) is the
5 quionoline and 4-chlorobenzoyl chloride is the acid halide. llle product, 1-(4-chlorobenzoyl)-2-
cyano-6,7-difluoro-1,2-dihydroquinoline, has a m.p. 184-185C.
Calcd for C17HgClF2N20: C, 61.74; H, 2.74; N, 8.47; Cl, 10.72; F, 11.49.
Found: C, 61.59; H, 2.82; N, 8.48; Cl, 10.6; F, 11.37.
ExamDle 25 1-(4-chlorobenzoyl)-6,8-difluoro-2-cyano-1,2-dihydroquinoline
Following the procedure of Exarnple la, 6,8-difluoroquinoline (Preparation 2) issubstituted for quinoline and 4-chlorobenzoyl chloride is used as the acid chloride. The
product, 1-(4-chlorobenzoyl)~,8-difluoro-2-cyano-1,2-dihydroquinoline, has a m.p. 164-165C.
Example 26 1 -(4-chlorobenzoyl)-7,8-difluoro-2-cyano-1,2-dihydroquinoline
Following the procedure of ExarDple la, 7,8-difluoroquinoline (Preparation 3) issubstituted for quinoline and 4-chlorobenzoyl chloride is used as the acid chloride m.p. 154-
lSgC.
Calcd: C, 72.72; H, 4.27; N, 8.48.
Found: C, 72,54; H, 4.35; N, 8.44.
~mEI~ 1-E-(3-(4-bromophenyl)-1-oxo-2-propenyl)-2-cyano-1,2~ihydroquinoline
Following the procedure of Example la, E4-bromocinnamoyl chloride is substituted for
the acid halide. The product, 1-E-(3-(4-bromophenyl)-1-oxo-2-propenyl)-2-cyano-1,2-
diDydroquinoline, has a m.p. 175-177C.
Example 28 1 -E-(3-(4-chlorophenyl)- 1 -oxo-2-propenyl)-2-cyano~,7-difluoro- 1,2-
dihydroquinoline
Following the procedure of ExaJnple la, 6,7-difluoroquinoline (Preparation 1) issubstituted for quinoline and 4-chlorocinnamoyl chloride is substituted for the acid halide. The
product, l-E-(3-(4-chlorophenyl)-1-oxo-2-propenyl)-2~yano~,7-difluoro-1,2-dihydroquinoline,
has a m.p. 97-98C.
Exarnple 29 1 -E-(3-(4-fluorophenyl)- 1 -oxo-2-propenyl)-2-cyano~,7-difluoro- 1,2-
dihydroquinoline
Following the procedure of Example la, 6,7-difluoroquinoline (Preparation 1) is
substituted for quinoline and 4-fluorocinnarnoyl chloride is used as the acid chloride. Ihe
prodùct, l-E-(3-(~fluorophenyl)-1-oxo-2-propenyl)-2-cyano~,7-difluoro-1,2-dihydroquinoline,
has a m.p. 131-132C.
Exam~le 30a 1-(4-chlorobenzoyl)-2-cyano~,7,8-trifluoro-1,2-dihydroquinoline
Following the procedure of ExaJnple la, 6,7,8-trifluoroquinoline is substituted for
SUBSTITUTE SHEE~
wo 92t1650~ ~ I 0 1 3 ~ ~ PCT/US92/0~746
-20- ~
quinoline and 4~hlorociMamoyl chloride is substituted from the acid chloride. ll~e product, 1-
(4~hlorobenzoyl)-2-cyano-6,7,8-trifluoro-1,2-dihydroquinoline, has a m.p. 159-160C.
Exam~le 30b 1 -E,E-( I -o~o-2,4-pentadienyl-5-phenyl)-2~yano-1,2-dihydroquinoline
Following the procedure of Exarnple la, styryl acrolyl chloride is substituted for the acid
5 halide. The title product is ob~ained.
IH NMR (300 MH2, CDC13): d 6.04 (dd, ~ = I l,15 Hz, IH), 6.35 (d, J= 15 Hz, IH),6.37 (dd, 1= 1,6 Hz, IH), 6.77 (d, J= 9Hz, IH), 6.82 (dd, J=11,15 Hz, IH), 7.22-7.37 (m,
7H), 7.43 (m, 2H), 7.62 (dd, 1= 11,15 Hz, IH).
E%am~le 31 1 -(4-trifluorobenzoyl)-2-cyano-3-dimethylamino- 1,2,3,4-tçtrahydroquinoline
To 500 mg (1.52 mmol) of 1-(4-trifluoromethylbenzoyl)-2~yano-1,2-dihydroquinoline is
added 20 ml of methylene dichloride and 3 ml of anhydrous dimethylarnine (the dimethylamine
is cooled to below -20C before carefully opening the sealed bottle). lbe reaction is allowed to
stir at ambient temperature for 48 h., filtered and the resulting methylene chloride solution
evaporated to dryness. The residue is distributed between methylene chloride and water, and
15 the organic phase washed twice with water. The organic phase is dried over anhydrous sodium
sulfate, filtered, and evaporated to dryness to give crude product. The crude product is
chromatographed using 35 percent ethyl acetate in hexane to give 1-(4-trifluorobeyl)-2-
cyano-34imethylamino-1,2,3,4-tetrahydroquinoline which slowly crystallizes upon standing
(m.p. 104-106C).
20 Example 32 1-(4~hlorobenzoyl)-2-cyano-1,2,3,4-tetrahydroquinoline.
To one gram of 1-(4~hlorobenzoyl)-2-cyano-1,24ihydroquinoline (3 4 mmol) in 100 ml
of ssæ ethanol is added 0.10 g of platinum oxide The reduction reaction proceeds under
hydrogen at atmospheric pressure 4 hours. The reaction is followed by l~C (35% ethyl
acetate/he~ane). The reaction is filtered and the resulting ethanol solution is concentrated in
25 vacuo; the crude solid is recrystallized from 5 ml of absolute ethanol to give the
tetrahydroquinoline (1-(4-chlorobenzoyl)-2-cyano-1,2,3,4-tetrahydroquinoline, 0.21 mg, m.p.
134-135C).
Calcd.: C, 68.78, H, 4.41; N, 9.44, Cl, 11.94.
Found: C, 68.66; H, 4.81; N, 9.16; Cl, 11.71.
30 E~ ample 33 1-Z-(I-oxo-3-phenyl-2-propenyl)-2-cyano-1,2-dihydroquinoline
a) Phenylpropiolic acid (K~cK Chemicals, 1.05 g, 7.2 mmol) is dissolved in 14 ml of
methylene chloride and cooled to 0C. Oxalyl chloride (0.63 ml) and dimethylformamide (140
~1) are added. The reaction is stirred at 0C for 30 min., then warmed to ambient temperature
and stirred an additional 3 hours. The reaction is concentrated in WCUO and further dried in
35 uacuo for 3 h. at ambient tçmperature. Quinoline (0.57 ml, 4.8 mmol) is dissolved in 14 m] of
methylene chloride and 0.42 ml of trimethylsilylcyanide is added. Ihis solution is added to the
SUBSTITUTE SHEET
, `. .
- . ...... -.
,
.,.~. .
. ~
wo 92/16508 2 1 ~ 1 3 3 5 PCr/US9t/01746
!-` -21- .
phenylpropiolic acid chloride prepared above. Aluminum trichloride (catalytic, 5 mol percent)
is added and the reac~ion is stirred overnight at ambient temperature. The reaaion is filtered
Lhrough a plug of silica gel and washed with 75 ml of methylene chloride. The organics are
washed with saturated aqueous sodium bicarbonate and evaporated in vacuo. Purification by
S flash column chromatography (25 percent ethyl acetate/hexane) provides 600 mg of the desired
product.
b) The above produa (200 mg) is dissolved in 7 ml of methanol and 22.6 mg of
palladium on calcium carbonate poisoned with lead (5% palladium, Aldrich) and 9 mg of
quinoline is added. The mixture is hydrogenated at 18 psi for 7.5 hours. Filtration of the
10 reaction and concentration in vacuo affords an oil which is dissolved in 95 percent ethanol and
chilled at 5C for 20 hours. The crystalline product is collected to provide l-(l-oxo-3-phenyl-
2-propenyl)-2-cyano-1,2-dihydroquinoline.
Calcd.: C, 79,70; H, 4.93; N, 9.79.
Found: C, 79.31; H, 5.01; N, 9.56.
15 Procedure fQr ~reparation Qf benzoxazineS~hart B!
Referring now to Chart B, figure 1, 1-(o-aminophenoxy)-2,2-diethoxyethane (I eq.) is
prepared following the procedure of F. Choiccara et al., Tetrahedron 32, 1407 (1976). The
diethoxyethanè is dissolved in, preferably ether, (0.1-0.3 M) and ethyl diisopropylamine (1-1.2
eq.) is added. The reactants are cooled to 0 C and one equivalent of the appropriate
20 ciMamoyl acid halide, preferably substituted c~nnamoyl acid chloride, is added dropwise. The
reaction is allowed to proceed for a time sufficient to allow as complete a reaction as possible,
e.g. 10 min. to I hour. The reactants are added to saturated sodium bicarbonate and ex~racted
with methylene chloride. The organic layers are combined, washed with water, dried over
anhydrous sodium sulfate, and concentrated in vacuo. The arnide product (Chart B, fig. 2) is
25 purified using flash chromatography in ethyl acetate/hexanes.
The arnide (Chart B. fig. 2) (I eq.) is dissolved in methylene chloride (0.4-0.5M) and the
reaction cooled to -20C. An acid, preferably boron trifluoride etherate, (1-1.2 eq.) is added
and the reaction mixture is stirred and slowly warrned to arnbient temperatures. Stirring
continues for from 12 to 20 hours. The mixrure is diluted with me~hylene chloride, washed
30 with saturated aqueous sodium bicarbonate, dried over anhydrous sodium sulfate, and
concentrated in vacuo. The product, a substituted 3,4~ihydro-3~thoxy-2H-1,4-benzoxazine
(Chart B, fig. 3), is purified by flash chromatography using a chloroform/methanol solvent.
The benzoxazine (Chart B, fig. 3, 1 eq.) is dissolved jD methyleDe chloride (0.1 -0.2M) and
I eq. (each) of azidotrimethylsilane and boron trifluoride etherate is added. ll~e reaction is
35 stirred at arnbient temperature for 3-24 h., diluted with methylene chloride, washed with
saturated sodium bicarbonate and again with saturated sodium chloride, dried over sodium
- SUBSTITUTE SHEE~
. ~ , ,
,
,
, ~ .
wo 92/1650X 2 ~ O 1 ~ ~ 5 -22- PCT/US92/01746
sulfate, and concentrated ~n wcuo Purification by nash chromatography in e~hylacetate/hexane
provides a substituted 3~4-dihydro-2H-1,4~ihydrobenzoxazine-3-azide (Chart B, fig. 4).
Fur~7er purification is accomplished by recrystallization in ethylene/hexane.
E~ample 34 4-(p-chlorocinnamoyl)-3,4-dihydro-2H- 1,4-benzoxazine-3-azide
a) 2-(4-chlorocinnamidinyl)phenoxy-1, I-diethoxyethane
1.24 g of 1-(o-aminophenoxy)-2,2-diethoxyethane (4.85 mmol), prepared following the
procedure of F Chioccara, F. Prota, R.H. Thompson, TetrahedrQn 32, 1407 (1976), is
dissolved in 40 ml of e~her and ethyl diisopropylarnine (5.82 mrnol, 0.75 g) is added. The
reaction is cooled to OC. p-Chlorocinnamoyl chloride (4.85 mmol) dissolved in 10 ml of
ether is added dropwise. After 10 min. of stirring, the reaction is poured in~o saturated
aqueous sodium bicarbonate and extracted with methylene chloride. The organic layers are
combined, washed with water, dried over anhydrous sodium sulfate, and concentrated in vacuo.
Purification via flash column chromatography (200 g silica gel, 10% ethyl acetate/hexane)
provides 2-(2-cblorocinnamidinyl)phenoxy)~ diethoxyethane.
b) 4-(p-chlorocinnamoyl)-3,4-dihydro-3-ethoxy-2H-1,4-benzoxazine
2-~2-~p~hlorocinnamidinyl)phenoxy-1,1-diethoxyethane (I mmol), syntbesized above, is
dissolved in 2 rnl of methylene cbloride and the reaction is cooled to -20C. E~oron trifluoride
etherate (1.2 mmol) is added, tbe reacsion allowed to slowly warm to room temperature over
approximately 2 hours and is then stirred overnight. The mixture is diluted witb methylene
chloride, washed with saturated aqueous sodium bicarbonate, dried over anhydrous sodium
sulfate, and concentrated in vacuo. Purification by flash column chromatogtaphy (100 g silica
gel, 100% chloroforrn) provides 4-(p-chlorocinnamoyl)-3,4-dihydro-3~thoxy-2H-1,4-
benzoxazine
c) 4-(p-chlorocinnamoyl)-3,4-dihydro-2H- 1,4-benzoxazine 3-azide
4-(p-Chlorocinnamoyl)-3,4-dibydro-3-ethoxy-2H-1,4-benzoxazine (1 m nol) synthesized in
step (b) is dissolved in 7 ml of methylene chloride Azidotrimethylsilane (1 mmol) and boron
trifluoride etherate (I mmol) are added. After stirring about 3 hours at room temperature, the
reaction is diluted with methylene chloride, washed with saturated sodium bicarbonate,
saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated in vacuo.
Purification by flash column chromatogtaphy (10% ethyl aceta~e/hexane) provides product
which is recrystallized from ethyl acetate/hexane to afford the desired product (~(p-
chlorociMamoyl)-3, dihydro-2H-1,4-benzoxazine-3-azide).
Procedure for ~re~aration of amidoximes (Chart C).
Referring now to Chart C, figure 1, the Reissert compound (0.68 eq.) is dissolved in
methanol at 30 to 50~C and slowly added to hydroxylamine in methanol at 0C. Thehydroxylamine is prepared by adding sodium metho%ide (10 ~q.) to hydroxylamine
SUBSTITUTE SHEET
..
.: :
wo 92/~6~08 2 1 0 1 3 3 ~ PCr/~'S92/0174b
23-
hydrochloride (I eq.) in methanol and then filtering. The Reissen-hydroxylarnine reaction is
allowed to warm to room temperature while stirring. It is then heated from 30C to reflux for 1
to 24 hours. The reaction mixture is cooled to room temperature, diluted with chloroform,
washed with saturated sodium bicarbonate, dried over sodium sulfate and concentrated in
S wcuo, The product (Chart C, fig. 2) is recrystallized from aqueous (95%) methanol.
As an alternative procedure we use the following:
Hydroxylamine hydrochloride (I eq.) is added to a stirred mixture of I eq. sodium
carbonate in 50 percent aqueous ethanol (0.6-0.8M). The reaction is diluted to 0.06-0.08M
with absolute ethanol. Then 0.8 eq. of the desired Reissen compound is added and the reaction
10 is heated at from 50C to reflux tempetature under Gitrogen for 2.5 to 24 hours. lbe reaction
poured is mixed into ice water and extracted with ethyl acetate or butanol. The organics are
concentrated in vacuo, dissolved in ethanol, and chilled. The solid product is collected and
dried.
The pharmacological salt of the amidoxime may be synthesized by use of an appropriate
15 acid. Examples of such salts include hydrochloride, hydrobromide, fumarate, maleate,
succinate, citrate, tosylate, and mesylate.
ExamDle 35 1-(~chlorobenzoyl)-2-carboximidamide-1,24ihydroquinoline
l-(~chlorobenzoyl)-2-cyano-1,24ihyd-oquinoline (0.2g, 0.68 mmol, see Popp, citedabove) is dissolved in warm methanol and added to a solution of hydroxylamine in methanol.
20 The hydroxylamine is prepared by the addition of sodium methoxide (0.232 mJ of 259G w/w
solution in methanol, I mmol) to 70 mg of hydroxylamine hydrochloride ~I mmol) in methanol
followed by filtration to remove precipitate. The Reissert-hydroxylamine reaction is stirred at
ambient temperature for 3 h and subsequently heated to 40C for I hour. The mixture is
evaporated to dryness, dissolved in 2 ml methanol, one drop of water added, and chilled to
25 5C. The solids are collected and washed with methanol to afford 1-(4-chlorobenzoyl)-2-
carboximidamide-1,2-dihydroquinoline, m.p. 160-161C.
In an analogous manner the following are synthesized by substituting the appropriate
Reissert compound.
Example 36a 1 -E-(3-(4-bromophenyl)- 1 -oxo-2-propenyl)-2~arboximidamide-1,2-
dihydroquinoline
Following the procedure of Example 35, 1-E-(3-(4-bromophenyl)-1-oxo-2-propenyl)-2-
cyano-1,2-dihydroquinoline is used as the Reissert compound. The product, I-E-(3-(4-
bromophenyl)- I -oxo-2-propenyl~2-car~oximidamide- I ,?-dihydroquinoline, has a m.p. 18
181C.
3s ExamD!e ~6~ 1-E-13-(4-bromophenyl)-1-oxo-2-propenyl]-N-(acetyloxy)-2-
quinolinecarboximidarnide
SUBSTITUTE SHEET
. . - . ~ ~ , - ` '.
.~ " . .:.,; .
. - -~.- . :
:: :
Wo 92J1650X PCI/US92/01746
2~ ~133~ -24- ~
The purified product of Example 36(a) is dissolved in 0.75 ml methylene chloride.
Pyridine (21 ,~11) is added and the reaction is cooled lo 0C. Acetic anhydride (25 ~LI) is added
and the reaction stirred 15 min. a~ 0C, and then brought to arnbient temperature. After I h.,
the reaction is diluted with methylene chloride, poured into saturated aqueous sodium
S bicarbonate, dried over sodium sulfate, and concentrated in vacuo. The residue is triturated
wilh 9596 aqueous ethanol and the solids collected to yield I E-13-(4-bromophenyl)-1-oxo-2-
propenyl]-N~acetyloxy)-2-quinolinecarbo%imidamide(m.p. 151-153C).
Example 37a 1 -E-(3-(4-chlorophenyl)- 1 -oxo-2-propenyl)-2-carboximidarnide- 1,2-dihydro-
quinoline
Following the procedure of Example 35, 1-E-(3-(4-chlorophenyl)-1-oxo-2-propenyl)-2-
cyano-1,2-dihydroquinoline is used as the Reissert compound. The product, I-E-(3-(4-
chlorophenyl)-l-oxo-2-propenyl)-2-carbo%imidamide-1,2-dihydroquinoline, has a m.p. 174-
175C.
Exam,~le 37b 1 -E-(3-(4-chlorophenyl)- 1 -oxo-2-propenyl)-2-carboximidamide- 1,2-dihydro-
quinoliDe, hydrochloride
The product of Example 37a, 1-E-(3-(4-chlorophenyl)-1-oxo-2-propenyl)-2-
carboximidamide-1,2-dihydroquinoline, is dissolved in methanol and chlorotrimethyl silane is
added. The addition of diethyl ether causes the desired salt to crystallize. 'Ihe crystals are
collected to afford l-E-(3-(4-chlorophenyl)-1-o%o-2-propenyl)-2-carbo~imidamide-1,2-dihydro-
quinoline, hydrochloride (m.p. IS8-160C).
Example 37c 1 -E-(3-(4-chlorophenyl)- 1 -oxo-2-propenyl)-2-carboximidamide- 1,2-dihydro-
quinoline, methane sulfonate
The product of E%ample 37a, 1-E-(3-(4-chlorophenyl)-1-oxo-2-propenyl~2-
carboximidamide-1,2-dihydroquinoline, is dissolved in methanol and methane sulfonic acid is
added. The volatiles are removed in vacuo and the residue recrystallized from d;ethyl
ether/chloroforrn. The crystals are collected and dried to yield l-E-(3-(4~hlorophenyl~1-oxo-
2-propenyl)-2-carbo%imidamide-1,2-dihydroquinoline, methane sulfonate (m.p. 139-140C).
Example 37d 1 -E-13-(4-chlorophenyl)- 1 -oxo-2-propenyl]-N-(acetyloxy)-2-quinoline-
carboximidarnide
Following the procedure of Example 36(b) and making non-critical variations, but starting
with the product of Example 37(a), 1-E-3-~4-chlorophenyl)-1-oxo-2-propenyl~2-
carboximidarnide-1,2-dihydroquinoline, the title compound is obtained. 1-E-[3-(4-chlorophenyl)-
I -oxo-2-propenyl]-N-(acetyloxy)-2-quinolinecarboximidarnide (m.p . 132- 133C).Example 37e 1-E-13-(~chlorophenyl)-1-oxo-2-propenyl~-N-methoxycarbonyloxy-2-
quir olinecarboximidamide
Following the general procedure as described for Examples 36(b) and rnal~ing non~ritical
SUBSTITUTE SHEE~
- . . ; . :
.
,
.
WO 92/16~08 21~1~ 3 5 PCr/~,'S92tO1746
f
-25 -
varia~ions, but staning with the product of E%ample 37ta), 1-E-(3-(4-chloropbenyl)-1-o%o-2-
propenyl)-2 carboximidamide-1,2-dihydroquinoline, and using methylchloroforrnate. the title
compound is obtained, I -E-~3-(4-chlorophenyl)- 1 -oxo-2-propenyll-N-methoxycarbonyloxy-2-
quinolinecarbo%imidamide (m.p. 173-174C).
E~amPIe 38 1-E-(3-(4-hydroxyphenyl)-1-oxo-2-propenyl)-2-carboximidamide-1,2-
dihydroquinoline
Sodium carbonate (0.31 g) is dissolved in 2 rnl of ethanol and 2 ml of water.
Hydro%ylamine hydrochloride (0.202 g) is added and the reaction is diluted with 40 mJ of
absolute ethanol. I-E-(3-(4-acetoxyphenyl)-1-o%o-2-propenyl)-2-cyano-1,2-dihydroquinoline
(Example 14, 0.80 g) is added and the reaction heated to reflux for 1.5 hours. After cooling, it
is poured into 400 ml of ice water, and extracted with chloroform (3 x 200 rnl.) After
concentration in vacuo, the residue is triturated with 3 mJ of methanol. The solids formed are
collected to afford 199 mg of the title compound (1-E-(3-(4-hydroxyphenyl~1-o%o-2-propenyl)-
2-carboximidamide- 1,2-dihydroquinol ine, m .p . 164- 166C) .
Example 39 1-(4-chlorobenzoyl)-8-fluoro-2-carboximidamide-1,2-dihydroquinoline
Following the general procedure according to Example 38 and making non-critical
variations, but using 1-(4-chlorobenzoyl)-2-cyano-8-fluoro-1,2-dihydroquinolu~e (Example 21)
the title compound is obtained (1-(4~hlorobenzoyl)-8-fluoro-2 carboxamidamide-1,2-
dihydroquirlolinc,m.p, 186-187C).
Exatncle 40 1-(4-chlorobenzoyl)~,7-difluoro-2 carboximidamide-1,2~ihydro-quinoline
Following Exa nple 38 and using 1-(4-chlorobenzoyl)~,7-difluoro-2-cyano-1,2-
dihydroquinoline (Example 24) the title compound is obtained (1-(4-chlorobenzoyl)~,7-
difluoro-2-carbo%irnidamide-1,2-dihydroquinoline,m.p. 128-129C).
Exarn~le 41 1-(~chlorobenzoyl)~fluoro-2-carboximidamide-1,2-dihydroquinoline
Following the general pro~edure of Example 38 and making non-critical variations, but
using 1-(4 chlorophenyl)-2-cyano~fluoro-1,2-dihydroquinoline (Exarnple 19), the title
compounD is obtained (1-(4-chlorobenzoyl)~fluoro-2~arboximidamide-1,2-dihydroquinoline,
m.p.162-163C).
Exarnple 42a 1 -E-(3-(4-cyanophenyl)- 1 -oxo-2-propenyl)-2-carboximidamide- 1,2- dihydroquinoline
Following ~e general procedure of Exarnple 35 and malcing non-critical variations, but
starting with l-E-(3-(4-cyanophenyl~-1-oxo-2-propenyl)-2-cyano-1,2-dihydroquinoline (Example
I l, 0.20 g), the title compound is obtained (1-E-(3-(~cyanophenyl)-1-oxo-2~ropenyl)-2-
carboximidamide-l ,2~ihydroquinoline, O. l lg, m.p. 205-206~C).
Example 42b 1-E-(3-(4-cyanopneDyl)-l-oxo-2-propenyl)-2-carboximidamide-1,2-
dihydroquinoline hydrochloride
- SUBSTITUTE SHEET
,: :
. ,--.
, . . .
. , .
wo 92/16508 ~ 3 3 5 - PCr/US92tol746
-26-
The product of E~ample 42a (1.0 g) is suspended in 10 ml of methanol and concentrated
hydrochloric acid (0.29 ml) is added. llle solution is diluted with ether until crystals formed
and then it is cooled to 5~C. The crystals are collected and dried to afford 0.68 g of the
hydrochloride salt (I-E-(3-(4-cyanophenyl)-1-oxo-2-propenyl)-2-carbo%imidamide-1,2-dihydro-
5 quinoline hydrochloride, m.p. 157-158C).
_olo~ical Activit~
The utility of the compounds of the invention is demonstrated by the ability of the
compounds used to practice the method claimed in this invention to inhibit viral reverse
transcriptase, an enzyme essential for human immunodeficiency virus replication. This enzyme
10 has characteristics which differentiate it from other known cellular polymerases and it is a
unique enzyme which is not found in uninfected cells. Viral reverse transcriptase is found in
e%traas from baaerial clones prepared according to the procedure described by Larder, B.,
Purifoy D.. Powell. K. and Darby, G., ~AIDS Virus Reverse Transcriptase Defined by High
Level E%pression in Escherichia coli.~, EMBO J. 6:3133-3137 (1987). Inhibition of this
enzyme is determined in a cell free assay which measures the level of radioactive precursors
incorporated into DNA. Extracts prepared according to the procedure of Kleid, D. G., et al.,
Science, 1125-1129 (1981) are incubated in a mixture of inbibitor, 20 mM dithiothreitol, 60
mM sodium chloride, 0.05% NP40, 10 mM magnesium chloride, 50 mM Tris pH 8,3, 10 ~LM
t35S]-labeled deo%ynucleoside-S'-triphosphate, 10 ~g/ml RNA template (poly rC or poly rG)
and 5 ~g/ml l)NA primer ~oligo dG or oligo dT) for 15 minutes at 37C. Incorporation of
radio labeled precursor is determined by sponing aliquots of the reaction mixture on DE 81
paper, washing the papers to remove unicorporated precursor, drying, and determining counts.
The results of various assays are combined and reported as % inhibition of reverse transcriptase
activity at a 100 f~M dose in Table I.
The utility of tbis invention is further demonstrated by the ability of various compounds
used to practice the method(s) claimed in this invention to inhibit HlV-induced syncytia
formation in a tissue culture assay using MT-2 cells infected wi~h HIV-I. l'his test is described
by Nara et al., ~Quantitative infectivity assay for HIV-I and -2", Nature 332: 469470, 1988,
as well as by Mariano Busso, et al., ~Nucleotide Dimers Suppress HIV Expression in utro" in
AIDS RESEARCH AND HUMAN RETROVIRUSES, vol. 4, No. 6, pages 449455 (1988),
Mary Ann Liebent, Inc., Publishers. The results (IC50 means the concentration, in ~M of
drug, required to inhibit syncytia formation to the extent of 509G) of various assays are
combined and report~d in Table I. In comparison, the known commercial compound, AZT,
e~hibited similar anti~HrV poteney in uhis assay with 100 percent and 50 percent reductions in
syncytia formation at concentrations of appro~imately 1 ~LM and 0.2 ~LM, respectively ~data not
shown).
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The utility of the compounds of the invenlion to practice the method claimed in this inven-
tion is further demonstrated by the activity of this compound in the inhibition of HIV infection
in primary peripheral blood Iymphocytes ~primary PBL assay). The primary PBL assay offers
the following advantages:
S (a) The assays are performed with primary human Iymphocytes. Thereby, undesired testing
of transformed cell lines is avoided in which host cell and virus may have undergone processes
of mutual adaptation. Performance of cell culture in serum containing media closely mimics the
in vivo situation. (b) The primary PBL assay distinguishes between uue antiviral effect which is
due to the drug and cytostatic/cytotoxic reactions. (c) Viral replication Is precisely followed by
kinetic measurement of viral nucleic acids and proteins. (d) Nucleic acids (totaJ HIV-RNA
inua- and extracellular) and protein (secreted p24) are measured in parallel which permits one
to differentiate between the compound's effect on virus replication and on the expression of
viral proteins. This leads to additional information regarding the efficacy of the test compound.
(e) Tolerance of the cell culture against low arnounts of organic solvents also permits the
investigation of hydrophobic substances. (f) The dose of the drug causing balf maximal
suppression of virus replication is deterrnined. (g) The screening system is standardized and
automated to a high degree.
The primary PBL assay uses the following procedure:
Effects of the compounds of the invention on cell proliferation are determined by
Iymphocyte proliferation assays. Starting witb a 100 micromolar solution, the compound is
serially diluted 10 fold. One tenth of the concentration of a compound causing half maximal
inbibition of cellular proliferation is employed for all subseque~t testing.
Peripheral human Iymphocytes are isolated by density gradient centrifugation. After
stimulation by mitogen the cells are infected with a standardized preparation of HIV.
Subsequently, the infected cells are cultured in the presence of the drug for four days. Indivi-
dual cultures are established to measure viral replication three and four days following
infection. Untreated cells and AZT-treated cells are included as controls in parallel with the
drugs under investigation.
The amount of viral core protein p24 synthesized and released by the infected cells is
determined in the supernatant by the capture-ELlSA technique on days three and four. By
comparing with a standard preparation, the arnount of protein produced by the virus
infected cells is quantified.
The total arnount of viral RNA synthesized by the; Ifected Iymphocytes is determined by a
special nucleic acid hybridization technique on days three and four of culture. By including a
standard preparation of HIV-RNA the amount of synthesized RNA is quantified.
If a drug shows antiviral effects in the primary assay, all steps of the primary assay are
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repeated, In addi~ion~ viability of HlV-infected cells is determined in parallel with assays for
viral p24 and RNA. In order to evaluate the half ma~imal antiviral effect of the drug, a
concentration dependency of the drug action is measured.
The compounds of the invention are assayed according to this procedure, The anti-HlV
5 acti1,dty, as measured by the inhibition of the release of core p24 protein in HIV infected human
Iymphocytes, is used to calculate antiviral EDso (the concentration required to give a 50%
reduction in p24 synthesis), Results are shown in Table 1,
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TABLE I
BIOASSAY RESULTSI
Syncytla PBL % RT Inhib
InhibitionED50 at
Example ED50(,UM) (JlM) 100 ~L
37a 0 I I _ 5~86
_ 35 0.1-1 77-89
7 <0.3 0.003 7~
l l <0.03 0.001-0.01 91-97
32 < 0.3 0.1-1 67-74
3 <0.3 0.1-1 50-74
8 0.1 0.1 75-91
2 0.3 _ 1-10 52-73
la l 0.1-1 69-72
6 0.1 0.1 60-82
17 <0.3 0.1-1 49-58
Ib2 3 10 3041
18 > 3 1-10 44~55
55-60
16 > 3 1-10 33-50
_
12 30 0.1 37-68
l9a 1-10 65-73
22 - 10 3549
21 l 62-76
24 _ I 60-85
9 1 - 10 58-62
K3 4 --10 39-78
> 3 - 10 18-61
23 18 - 10 2740
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TABLE I
BIOASSAY RESULTS
(Continued)
r~ - Syncytia PBL % RT lnb
Inhibition EDSo at
lE~arnple EDso (~M) M) _100 /uM
I 10 0.3-2.8 I 42{i0
28 < I 78-85
14a I 54 66
¦ 14b < 0.29 0.1-1 64-87
1 13 <3 0.1 48-7~
1 33 10 31-S5
< 10 53-67
26 < 10 37-68
¦ 36a 0.1-1 65-76
l 27 <0.27 0.01 K71
¦ 42a 52-56
1 41 81-85
¦ 37e ~ 0.1-1 18-42
¦ 37d 2.5 0.010.1 17-28
~ 36b 0.1-1 35-53
25 r 3~ 1-10 69-70
1 31 _1 11-36
30 1 - Assays are e~plained in specification.
2 - Known compound.
3 - Known compound: 1-(4-toluoyl)-2~yano-1,2-dihydroquinoline.
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