U.S. patent application number 10/725657 was filed with the patent office on 2004-06-17 for compounds and methods for inhibition of hiv and related viruses.
This patent application is currently assigned to Medivir AB. Invention is credited to Lind, Peter Thomas, Morin, John Michael, Noreen, Rolf, Ternansky, Robert John.
Application Number | 20040116418 10/725657 |
Document ID | / |
Family ID | 24405987 |
Filed Date | 2004-06-17 |
United States Patent
Application |
20040116418 |
Kind Code |
A1 |
Lind, Peter Thomas ; et
al. |
June 17, 2004 |
Compounds and methods for inhibition of HIV and related viruses
Abstract
The present invention is directed to compounds, compositions and
methods of inhibiting and treating HIV and related viruses, and
methods for cocrystallizing reverse transcriptase in vitro,
inhibition of the replication of HIV and related viruses
thereof.
Inventors: |
Lind, Peter Thomas;
(Huddinge, SE) ; Noreen, Rolf; (Huddinge, SE)
; Morin, John Michael; (Brownsburg, IN) ;
Ternansky, Robert John; (Carlsbad, CA) |
Correspondence
Address: |
BIRCH STEWART KOLASCH & BIRCH
PO BOX 747
FALLS CHURCH
VA
22040-0747
US
|
Assignee: |
Medivir AB
Huddinge
SE
|
Family ID: |
24405987 |
Appl. No.: |
10/725657 |
Filed: |
December 1, 2003 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
10725657 |
Dec 1, 2003 |
|
|
|
10076163 |
Feb 13, 2002 |
|
|
|
10076163 |
Feb 13, 2002 |
|
|
|
09567857 |
May 9, 2000 |
|
|
|
6376492 |
|
|
|
|
09567857 |
May 9, 2000 |
|
|
|
09114935 |
Jul 14, 1998 |
|
|
|
09114935 |
Jul 14, 1998 |
|
|
|
08601030 |
May 3, 1996 |
|
|
|
5849769 |
|
|
|
|
08601030 |
May 3, 1996 |
|
|
|
PCT/US94/09406 |
Aug 24, 1994 |
|
|
|
Current U.S.
Class: |
514/227.8 ;
514/235.2; 514/252.03; 514/252.04; 514/254.02; 514/326; 514/362;
514/370; 514/372 |
Current CPC
Class: |
A61K 31/44 20130101;
C07D 231/40 20130101; C07D 239/42 20130101; C07D 277/56 20130101;
C07D 401/12 20130101; C07D 277/48 20130101; C07D 213/85 20130101;
C07D 277/82 20130101; C07D 213/76 20130101; C07D 237/20 20130101;
A61K 31/40 20130101; C07D 213/75 20130101; A61K 31/50 20130101;
A61K 31/17 20130101 |
Class at
Publication: |
514/227.8 ;
514/235.2; 514/254.02; 514/252.03; 514/252.04; 514/362; 514/370;
514/372; 514/326 |
International
Class: |
A61K 031/541; A61K
031/5377; A61K 031/496; A61K 031/454 |
Claims
1. A method for treating HIV which comprises administering to a
patient in need thereof, an effective anti-HIV amount of a compound
of the formula 37wherein A is 38where Z.sub.i is O, R.sub.1 is
isothiazolyl, substituted isothiazolyl, thiazolyl, substituted
thiazolyl, thiadiazolyl, substituted thiadiazolyl; R.sub.2 is a
group of the formula 39wherein R.sub.5 is a stable, saturated or
unsaturated, substituted or unsubstituted 3 to 8 member organic
monocyclic ring having 0 to 4 heteroatoms selected from S, O and N;
or R.sub.5 is a stable, saturated or unsaturated, substituted or
unsubstituted 7 to 10 membered organic bicyclic ring having 0 to 5
heteroatoms selected from S, O or N; and are; R.sub.3 and R.sub.4
are hydrogen; or a pharmaceutically acceptable salt thereof.
2. The method of claim 1 wherein R.sub.5 is
cyclo(C.sub.3-C.sub.8)alkyl, cyclo (C.sub.3-C.sub.8) alkenyl;
isothiazolyl, substituted isothiazolyl, tetrazolyl, substituted
tetrazolyl, triazolyl, substituted triazolyl, pyridyl, substituted
pyridyl, imidazolyl, substituted imidazolyl, phenyl, substituted
phenyl, naphthyl, substituted naphthyl, benzoxazolyl, substituted
benzoxazolyl, benzimidazolyl, substituted benzimidazolyl,
thiazolyl, substituted thiazolyl, oxazolyl, substituted oxazolyl,
benzothiazolyl, substituted benzothiazolyl, pyrazinyl, substituted
pyrazinyl, pyridazinyl, substituted pyridazinyl, thiadiazolyl,
substituted thiadiazolyl, benzotriazolyl, substituted
benzotriazolyl, pyrrolyl, substituted pyrrolyl, indolyl,
substituted indolyl, benzothienyl, substituted benzothienyl,
thienyl, substituted thienyl, benzofuryl, substituted benzofuryl,
furyl, substituted furyl, quinolinyl, substituted quinolinyl,
isoquinolinyl, substituted isoquinolinyl, pyrazolyl, and
substituted pyrazolyl.
3. The method of claim 1, wherein R.sub.1 is, thiazolyl or
substituted thiazolyl
4. The method of claim 1 wherein; R.sub.1 is thiazolyl,
(4-methyl)thiazolyl, (4,5-dimethyl)thiazolyl, (4-cyano)thiazolyl,
(4-ethyl)thiazolyl, 4-(3-pyridyl)thiazolyl,
4-(3-nitrophenyl)thiazolyl, 1,3,4-thiadiazolyl,; 40R.sub.5 is
phenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl,
2-ethoxyphenyl, 2-methylphenyl, 3-methylphenyl, 2-fluorophenyl,
2,6-difluorophenyl, 2-fluoro-6-methoxyphenyl,
2-fluoro-6-ethoxyphenyl, 2,3,5,6-tetrafluorophenyl, 2-chlorophenyl,
3-chlorophenyl, 1-cyclohexenyl, 2-naphthyl, 2,5-dimethoxyphenyl,
2-azidophenyl, 2,3,4-trifluorophenyl, 2-fluoro-6-chlorophenyl,
2,6-dimethoxyphenyl, 2,3,6-trichlorophenyl, 2,6-dichlorophenyl,
2,3,5-trichlorophenyl, 3,5-dichlorophenyl, 3-fluorophenyl,
2,4-dimethoxyphenyl, 2-pyridyl, 2-(6-methoxy)pyridyl,
2-(6-ethoxy)pyridyl, 2-(6-fluoro)pyridyl, 2-(5-fluoro)pyridyl,
2-(4-fluoro)pyridyl, 2-(3-fluoro)pyridyl, 2-(6-chloro)pyridyl,
2-(5-chloro)pyridyl, 2-(4-chloro)pyridyl, 2-(3-chloro)pyridyl,
2-(5-methoxy-6-fluoro)pyridyl, 2-(3-methoxy-6-fluoro)pyridyl,
2-(6-methoxy-3-fluoro)pyridyl, 2-(5-ethoxy-6-fluoro)pyridyl,
2-(3-ethoxy-6-fluoro)pyridyl, 2-(6-ethoxy-3-fluoro)pyridyl,
2-(5,6-difluoro)pyridyl, 2-(3,6-difluoro)pyridyl,
2-(5,6-dichloro)pyridyl, 2-(3,6-dichloro)pyridyl- ,
2-(6-methoxy)pyridyl, 2-(6-ethoxy)pyridyl, 2-(1,3-pyrimidyl),
2-pyrazinyl, 3-pyridazinyl, 2,6-difluoro-3-methoxyphenyl,
2,6-difluoro-3-ethoxyphenyl, 2,6-difluoro-4-methoxyphenyl,
2,6-difluoro-4-ethoxyphenyl, 2-(3-ethoxy)pyridyl,
2-(3-methoxy)pyridyl, 2,6-difluorophenyl,
2,6-difluoro-3-N-methylcarboxamidephenyl, 2-fluoro-6-chlorophenyl,
3-bromo-6-methoxyphenyl, 3-ethoxyphenyl, 3-bromo-6-ethoxyphenyl,
3-(2-fluoro)pyridyl, (2-vinyl)phenyl, (3-vinyl)phenyl,
(3-methoxycarbonyl)phenyl, 5,6-dimethylbenzotriazolyl,
2,3-difluoro-6-methoxyphenyl, 2,6-difluoro-3-cyanophenyl,
3-ethynylphenyl, and 2,5-diethoxyphenyl.
5. The method as recited in claim 1 further comprising
administering at least one other anti-HIV agent to said
patient.
6. The method as recited in claim 5 wherein said agent is selected
from ddI, ddC, or AZT.
7. A compound having the formula 41wherein A is 42Z.sub.i is O;
R.sub.1 is isothiazolyl, substituted isothiazolyl, thiazolyl,
substituted thiazolyl, thiadiazolyl, substituted thiadiazolyl,
R.sub.2 is a group of the formula 43wherein R.sub.5 is a stable,
unsaturated, substituted or unsubstituted i) 3 to 8 membered
monocyclic ring having 0 to 4 hetero atoms or ii) a 7 to 10
membered bicyclic ring having 0 to 5 hetero atoms, said hetero
atoms being selected from S, O and N; and R.sub.3 and R.sub.4 are
hydrogen; or a pharmaceutically acceptable salt thereof.
8. The compound of claim 7 wherein the substituted R.sub.1 and/or
R.sub.5 groups have single or multiple substituents independently
selected from halo, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.5 alkoxy,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.2-C.sub.8
alkenoxy, amino, nitro, cyano, carboxy, hydroxymethyl, aminomethyl,
carboxymethyl, C.sub.1-C.sub.4 alkylthio, hydroxy, C.sub.1-C.sub.4
alkanoyloxy, carbamoyl, halo-substituted C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkoxy-substituted C.sub.1-C.sub.6 alkyl, a group
of the formula--SO.sub.2R.sub.xwherein R.sub.x is C.sub.1-C.sub.6
alkyl or amino; or a group of the formula 44wherein R.sub.x is
C.sub.1-C.sub.6 alkyl.
9. The compound of claim 7 wherein R.sub.1 is thiazolyl,
(4-methyl)thiazolyl, (4,5-dimethyl)thiazolyl, (4-cyano)thiazolyl,
(4-ethyl)thiazolyl, 4-(3-pyridyl)thiazolyl,
4-(3-nitrophenyl)thiazolyl, or 1,3,4-thiadiazolyl.
10. The compound of claim 7, wherein R.sub.5 is
cyclo(C3-C-8)alkenyl, thiazolyl, substituted thiazolyl, tetrazolyl,
substituted tetrazolyl, triazolyl, substituted triazolyl, pyridyl,
substituted pyridyl, imidazolyl, substituted imidazolyl, phenyl,
substituted phenyl, naphthyl, sustituted naphthyl, benzoxazolyl,
substituted benzoxazolyl, benzimidazolyl, substituted
benzimidazolyl, thiazolyl, substituted thiazolyl, oxazolyl,
substituted oxazolyl, benzothiazolyl, substituted benzothiazolyl,
pyrazinyl, substituted pyrazinyl, pyridazinyl, substituted
pyridazinyl, thiadaizolyl, substituted thadiazolyl, benzotriazolyl,
substituted benzotriazolyl, pyrrolyl, substituted pyrrolyl,
indolyl, substituted indolyl, benzothienyl, substituted
benzothienyl, thienyl, substituted theinyl, benzofuryl, substituted
benzofuryl, furyl, substituted furyl, quinolinyl, substituted
quinolinyl, isoquinolinyl, subsituted isoquinolinyl, pyrazolyl, and
substituted pyrazolyl.
11. The compound of claim 10, wherein R.sub.5 is phenyl,
2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2-ethoxyphenyl,
2-methylphenyl, 3-methylphenyl, 2-fluorophenyl, 2,6-difluorophenyl,
2-fluoro-6-methoxyphenyl, 2-fluoro-6-ethoxyphenyl,
2,3,5,6-tetrafluorophenyl, 2-chlorophenyl, 3-chlorophenyl,
1-cyclohexenyl, 2-naphthyl, 2,5-dimethoxyphenyl, 2-azidophenyl,
2,3,4-trifluorophenyl, 2-fluoro-67-chlorophenyl,
2,6-dimethoxyphenyl, 2,3,6-trichlorophenyl, 2,6-dichlorophenyl,
2,3,5-trichlorophenyl, 3,5-dichlorophenyl, 3-fluorophenyl,
2,4-dimethoxyphenyl, 2-pyridyl, 2-(6-methoxy)pyridyl,
2-(6-ethoxy)pyridyl, 2-(6-fluoro)pyridyl, 2-(5-fluoro)pyridyl,
2-(4-fluoro)pyridyl, 2-(3-fluoro)pyridyl, 2-(6-chloro)pyridyl,
2-(5-chloro)pyridyl, 2-(4-chloro)pyridyl, 2-(3-chloro)pyridyl,
2-(5-methoxy-6-fluoro)pyridyl, 2-(3-methoxy-6-fluoro)pyridyl,
2-(6-methoxy-3-fluoro)pyridyl, 2-(5-ethoxy-6-fluoro)pyridyl,
2-(3-ethoxyl-6-fluoro)pyridyl, 2-(6-ethoxy-3-fluoro)pyridyl, 2-(5,
6-difluoro)pyridyl, 2-(3,6-difluoro)pyridyl,
2-(5,6-dichloro)pyridyl, 2-(3,6-dichloro)pyridyl- ,
2-(6-methoxy)pyridyl, 2-(6-ethoxy)pyridyl, 2-(1,3-pyrimidyl),
2-pyrizinyl, 3-pyridazinyl, 2,6-difluoro-3-methoxyphenyl,
2,6-difluoro-3-ethoxylphenyl, 2,6-difluoro-4-methoxyphenyl,
2,6-difluoro-4-ethoxyphenyl, 2-(3-ethoxy)pyridyl,
2-(3-methoxy)pyridyl, 2,6-difluorophenyl,
2,6-difluoro-4-ethoxyphenyl, 2-(3-ethoxy)pyridyl,
2-(3-methoxy)pyridyl, 2,6-difluorophenyl,
2,6-difluoro-3-N-methylcarboxam- idephenyl. 2-fluoro-6-chlorphenyl,
3-bromo-6-methoxyphenyl, 3-ethoxyphenyl, 3-bromo-6-ethoxyphenyl,
3-(2-fluoro)pyridyl, (2-vinyl)phenyl, (3-vinyl)phenyl,
(3-methoxycarbonyl)phenyl, 5,6-dimethylbenzotriazolyl,
2,3-difluoro-6-methoxyphenyl, 2,6-difluoro-3-cyanophenyl,
3-ethynylphenyl or 2,5-diethoxyphenyl.
12. The compound of claim 7, wherein the N' linkage to R.sub.1 is
at the 2 position relative to a heteroatom in said isothiazolyl,
substituted isothiazolyl, thiazolyl, substituted thiazolyl,
thiadiazolyl, substituted thiadiazolyl.
13. A pharmaceutical formulation comprising an effective amount of
a compound as defined in claim 7; and a pharmaceutically acceptable
carrier or diluent therefor.
14. A pharmaceutical formulation according to claim 13, wherein
said agent is selected from ddI, ddC or AZT.
15. A method for treating or inhibiting HIV, comprising
administering to a patient suffering from HIV infection an amount
of a compound of claim 7 effective for treating or inhibiting HIV.
Description
[0001] This application is a Divisional of co-pending application
Ser. No. 10/076,163, filed on Feb. 13, 2003, which is a divisional
of co-pending application Ser. No. 09/567,857, filed on May 9, 2000
which is a continuation of application Ser. No. 09/114,935, filed
on Jul. 14, 1998, which is a divisional of application Ser. No.
08/601,030, filed on May 3, 1996, the entire contents of which are
hereby incorporated by reference and for which priority is claimed
under 35 U.S.C. .sctn. 120. Application Ser. No. 08/601,030 is the
national phase of PCT International Application No. PCT/US94/09406
filed on Aug. 24, 1994 under 35 U.S.C. .sctn. 371. The entire
contents of each of the above-identified applications are hereby
incorporated by reference.
FIELD OF THE INVENTION
[0002] The present invention relates to compounds and
pharmaceutically acceptable salts thereof and methods for treating
infections by HIV and related viruses and/or the treatment of
Acquired Immune Deficiency Syndrome (AIDS).
[0003] Also disclosed are pharmaceutical compositions containing
the compounds and the method of use of the present compounds alone
or in combination with other agents, for the treatment and
inhibition of AIDS and viral infection from HIV.
BACKGROUND OF THE INVENTION
[0004] A retrovirus designated Human Immunodeficiency Virus (HIV)
is believed to be the causative agent of the complex disease termed
Acquired Immune Deficiency Syndrome (AIDS) and is a member of the
lentivirus family of retroviruses (M. A. Gonda, F. Wong-Staal NR.
C. Gallo, "Sequence Homology and Morphological Similarity of HTLV
III and Visna Virus, A Pathogenic Lentivirus", Science, 227, 173,
(1985); and P. Sonigo and N. Alizon, et al., "Nucleotide Sequence
of the Visna Lentivirus: Relationship to the AIDS Virus", Cell, 42,
369, (1985)).
[0005] The HIV virus (also referred to as the AIDS virus) was
previously known as or referred to as LAV, HTLV-III, or ARV, and is
now designated by HIV-1. Other closely related variants of HIV-1
include HIV-2 and SIV (simian immunodeficiency virus), and mutants
thereof. The complex disease AIDS includes progressive destruction
of the immune system and degeneration of the central and peripheral
nervous system. The HIV virus appears to preferentially attack
helper T-cells (T-lymphocytes or OKT4-bearing T-cells) and also
other human cells, e.g., certain cells within the brain. The helper
T-cells are invaded by the virus and the T-cell becomes an HIV
virus producer. The helper T-cells are quickly destroyed and their
number in the human being is depleted to such an extent that the
body's B-cells as well as other T-cells normally stimulated by
helper T-cells no longer function normally or produce sufficient
lymphokines and antibodies to destroy the invading virus or other
invading microbes.
[0006] While the HIV virus does not necessarily cause death per se,
it does cause the human's immune system to be so severely depressed
that the human falls prey to various other diseases such as herpes,
Pneumocystis carinii, toxoplasmosis, cytomegalovirus, Kaposi's
sarcoma, and Epstein-Barr virus related lymphomas among others.
These secondary infections are separately treated using other
medications as is conventional. Early during infection, humans with
HIV virus seem to live on with little or no symptoms, but have
persistent infections. Later in the disease, humans suffer mild
immune system depression with various symptoms such as weight loss,
malaise, fever, and swollen lymph nodes. These syndromes have been
called persistent generalized lymphadenopathy syndrome (PGL) and
AIDS related complex (ARC) and develop into AIDS.
[0007] In all cases, those infected with the AIDS virus are
believed to be persistently infective to others. Further, AIDS and
AIDS related complex is after some time fatal.
[0008] A description of the mechanism by which the virus infects
its host is given in an article by R. Yarchoan, and S. Broder,
"Development of Antiretroviral Therapy for the Acquired
Immunodeficiency Syndrome and Related Disorders", New England
Journal of Medicine, 316, 557-564 (Feb. 26, 1987).
[0009] Considerable efforts are being directed toward the control
of HIV by means of inhibition of the reverse transcriptase of HIV,
required for replication of the virus. (V. Merluzzi et al.,
"Inhibition of the HIV-1 Replication by a Nonnucleoside Reverse
Transcriptase Inhibitor", Science, 25, 1411 (1990)). For example, a
currently used therapeutic compound, AZT, is an inhibitor of the
viral reverse transcriptase (U.S. Pat. No. 4,724,232).
Unfortunately, many of the now used compounds suffer from toxicity
problems, lack of bioavailability or are short lived in vivo, viral
resistance, or combinations thereof. Therefore, new compounds are
being investigated. For example, a PCT application published on
Feb. 18, 1993, (WO 93/03022; International Application Number
PCT/SE92/00533) discloses thiourea compounds which show promise in
the treatment and inhibition of HIV and AIDS.
[0010] Certain compounds falling within the scope of the definition
of the methods of the present invention have been published in
roles other than for the treatment of HIV. These compounds are
excluded by proviso from the compound claims. The Chemical Abstract
registry numbers or sources for these compounds are as follows:
[0011] a) i) 87977-01-3;
[0012] ii) 75808-56-9;
[0013] iii) 78329-62, 1128227-56-7, -49-8, 51-2, -48, 7, -48-6,
-50-1 and -46-5;
[0014] iv) 65094-08-8;
[0015] v) 39960-40-2 and -10-6, 64442-48-2 and -47-3;
[0016] vi) U.S. Pat. No. 3,705,903;
[0017] vii) 85180-12-7;
[0018] viii) 55474-82-3;
[0019] ix) 49551-06-6, -08-8, -08-4, -81-7, -67-9, -65-7, -58-8,
-31-7, -63-3 and -60-2, and 49552-03-6;
[0020] x) U.S. Pat. No. 4,057,636;
[0021] b) 100973-52-2, 92852-25-0;
[0022] c) 141403-37-4
[0023] Therefore, it is an object of the invention to provide
compounds and pharmaceutically acceptable salts thereof to inhibit
and/or treat HIV and AIDS.
[0024] Another object of the present invention is to provide
therapeutic formulations that are of value in the inhibition and/or
treatment of infection by HIV and the treatment or inhibition of
the acquired immune deficiency syndrome.
[0025] Another object is to provide methods for the inhibition
and/or treatment of infection by HIV and the resulting acquired
immune deficiency syndrome.
[0026] Other objects, features, and advantages will become apparent
to those skilled in the art from the following description and
claims.
DESCRIPTION OF THE INVENTION
[0027] The present invention provides compounds useful for the
inhibition and/or treatment of HIV and AIDS, either as compounds,
pharmaceutically acceptable salts, pharmaceutical composition
ingredients, whether or not in combination with other anti-virals,
immunomodulators, antibiotics, or vaccines. Methods of treating or
inhibiting AIDS, methods of inhibiting replication of HIV, and
methods of treating or inhibiting HIV in humans are also
disclosed.
[0028] The compounds used in the methods of the present invention
are those of the formula (IA) below 1
[0029] wherein A is 2
[0030] --C-- or --S--; and
[0031] Z.sub.i is O, Se, NR.sup.a or C(R.sup.a).sub.2, and
[0032] Z.sub.ii is --O or (.dbd.O).sub.2;
[0033] wherein R.sup.a is H, OR.sup.b, CN, NO.sub.2,
N(R.sup.b).sub.2, SR.sup.b, SO.sub.2R.sup.b,
SO.sub.2N(R.sup.b).sub.2, COR.sup.b, CO.sub.2R.sup.b,
CON(R.sup.b).sub.2, PO(R.sup.b).sub.2, PO(OR.sup.b).sub.2,
PO(NR.sup.b).sub.2, wherein R.sup.b is hydrogen, C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 substituted alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 substituted alkenyl, C.sub.2-C.sub.8 alkynyl,
C.sub.2-C.sub.8 substituted alkynyl, C.sub.1-C.sub.6 alkoxy,
C.sub.1-C.sub.6 substituted alkoxy, C.sub.4-10 aralkyl, C.sub.1-10
alkaryl, C.sub.1-10 alkylthio, C.sub.4-10 aralkylthio, C.sub.1-10
alkylsulfinyl, C.sub.4-10 aralkylsulfinyl, C.sub.1-10
alkylsulfonyl, C.sub.4-10 aralkylsulfonyl, carboxy, C-.sub.1-10
alkylthiocarbonyl, C.sub.4-10 aralkylcarbonyl, C.sub.4-10
aralkylthiocarbonyl, C.sub.4-10 aralkoxycarbonyl, C.sub.4-10
aralkoxycarbonyl, C.sub.1-4 alkyl, C.sub.4-10 aralkoxy, C.sub.1-12
dialkylamino-C.sub.1-6 aralkanoylamino C.sub.4-10 aralkylamino or
C.sub.1-C.sub.4 alkanoyloxy;
[0034] R.sub.1 is a stable saturated or unsaturated, substituted or
unsubstituted, 3 to 8 membered organic monocyclic ring having 0 to
4 hetero atoms selected from S, O, and N; or R.sub.1 is a stable,
saturated or unsaturated, substituted or unsubstituted, 7 to 10
membered organic bicyclic ring having 0 to 5 hetero atoms selected
from S, O, and N;
[0035] R.sub.2 is a group of the formula 3
[0036] wherein R.sub.5 is R.sub.1 as defined above; or R.sub.5 is a
group of the formula
(R.sub.10).sub.y--X--
[0037] wherein y is 1 or 2; X is N, S, O and R.sub.10 is R.sub.1 as
defined; or R.sub.10 is hydrogen, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, or C.sub.2-C.sub.6 alkynyl, substituted
C.sub.1-C.sub.6 alkyl, substituted C.sub.2-C.sub.6 alkenyl, or
substituted C.sub.2-C.sub.6 alkynyl; or R.sub.5 is hydrogen, halo,
cyano, carboxy, amino, thio, hydroxy, C.sub.1-C.sub.6 alkoxy,
C.sub.1-C.sub.6 substituted alkoxy, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, or
C.sub.2-C.sub.8 alkenoxy;
[0038] R.sub.6, R.sub.7, R.sub.8, and R.sub.9 are independently
C.sub.3-C.sub.8 cycloalkyl, hydrogen, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, substituted
C.sub.1-C.sub.6 alkyl, substituted C.sub.2-C.sub.6 alkenyl, or
substituted C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 substituted
alkoxy, halo, amino, nitro, cyano, C.sub.1-C.sub.5 alkoxy, hydroxy,
carboxy, hydroxymethyl, aminomethyl, carboxymethyl, C.sub.1-C.sub.4
alkylthio, C.sub.1-C.sub.4 alkanoyloxy, carbamoyl, or a halo
substituted C.sub.1-C.sub.6 alkyl; or two of which, along with the
carbons to which they are attached, combine to form a stable,
saturated or unsaturated, substituted or unsubstituted, 3 to 7
membered organic monocylic ring having 0 to 4 hetero atoms selected
from S, O, or N; or R.sub.6 and R.sub.8, or R.sub.7 and R.sub.9,
along with the carbon to which they are attached, form a stable,
saturated or unsaturated, substituted or unsubstituted, 3 to 7
membered organic monocylic ring having 0 to 4 hetero atoms selected
from S, O, or N;
[0039] R.sub.3 and R.sub.4 are independently hydrogen, hydroxy,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6
alkynyl, substituted C.sub.1-C.sub.6 alkyl, substituted
C.sub.2-C.sub.6 alkenyl, or substituted C.sub.2-C.sub.6 alkynyl,
substituted alkoxy, amino, cyano, nitro, C.sub.1-C.sub.6 alkoxy,
C.sub.1-C.sub.6 substituted alkoxy, carboxy, hydroxymethyl,
aminomethyl, carboxymethyl, C.sub.1-C.sub.4 alkylthio,
C.sub.1-C.sub.4 alkanoyloxy, halo-substituted
(C.sub.1-C.sub.6)alkyl, or carbamoyl;
[0040] or salts thereof;
[0041] It should also be understood that when the term "HIV" is
used, it includes HIV-1, components, mutant variations, subtypes,
and serotypes thereof, and related viruses, components, mutant
variations, subtypes, and serotypes thereof. When the term
"inhibit" is used, its ordinary meaning is intended, which is to
prohibit, hold in check, or discourage, and is not to be construed
to be limited to a particular process, procedure, or mechanism of
action.
[0042] The terms "stable, saturated or unsaturated, substituted or
unsubstituted, 3 to 8 membered", or "3 to 7 membered organic
monocyclic ring having 0 to 4 hetero atoms selected from S, O, and
N" include those wherein the nitrogen and sulfur hetero atoms are
optionally oxidized, and the nitrogen hetero atom optionally
quaternized. The substituted ring may have 1-8 substituents
independently selected from aryl, substituted aryl, halo,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.5 alkoxy, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.8 alkynyl, C.sub.2-C.sub.8 alkenoxy, amino,
nitro, cyano, carboxy, hydroxymethyl, aminomethyl, carboxymethyl,
C.sub.1-C.sub.4 alkylthio, hydroxy, C.sub.1-C.sub.4 alkanoyloxy,
carbamoyl, halo-substituted C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
alkoxy-substituted C.sub.1-C.sub.6 alkyl, a group of the
formula
--SO.sub.2R.sub.X
[0043] wherein R.sub.X is C.sub.1-C.sub.6 alkyl, aryl, substituted
aryl, or amino; or a group of the formula 4
[0044] wherein R.sub.X is as defined above.
[0045] The term "stable, saturated or unsaturated, substituted or
unsubstituted, 7 to 10 membered organic bicyclic rings having 0 to
5 hetero atoms selected from S, O, and N" includes those wherein
the nitrogen and sulfur hetero atoms are optionally oxidized, and
the nitrogen hetero atom(s) optionally quaternized. The bicyclic
rings may be substituted 1 to 8 times, the substituents
independently selected from those above listed for the monocyclic
rings.
[0046] Examples of such monocyclic and bicyclic rings are
cyclo(C.sub.3-C.sub.8)alkyl, cyclo(C.sub.3-C.sub.8)alkenyl;
isothiazolyl, substituted isothiazolyl, tetrazolyl, substituted
tetrazolyl, triazolyl, substituted triazolyl, pyridyl, substituted
pyridyl, imidazolyl, substituted imidazolyl, phenyl, substituted
phenyl, naphthyl, substituted naphthyl, benzoxazolyl, substituted
benzoxazolyl, benzimidazolyl, substituted benzimidazolyl,thiazolyl,
substituted thiazolyl, oxazolyl, substituted oxazolyl,
benzothiazolyl, substituted benzothiazolyl, pyrazinyl, substituted
pyrazinyl, pyridazinyl, substituted pyridazinyl, thiadiazolyl,
substituted thiadiazolyl, benzotriazolyl, substituted
benzotriazolyl, pyrrolyl, substituted pyrrolyl, indolyl,
substituted indolyl, benzothienyl, substituted benzothienyl,
thienyl, substituted thienyl, benzofuryl, substituted benzofuryl,
furyl, substituted furyl, quinolinyl, substituted quinolinyl,
isoquinolinyl, substituted isoquinolinyl, pyrazolyl, and
substituted pyrazolyl. Other examples of such ring systems may be
found in J. Fletcher, O. Dermer, R. Fox, Nomenclature of Organic
Compounds, pp. 20-63 (1974), and in the Examples herein.
[0047] The term "C.sub.1-C.sub.6 alkyl" includes such groups as
methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, t-butyl,
n-pentyl, n-hexyl, 3-methylpentyl, and the like.
[0048] The term "halo" and "halogen" refer to chloro, bromo,
fluoro, and iodo.
[0049] "C.sub.1-C.sub.6 alkoxy" refers to those groups such as
methoxy, ethoxy, propoxy, t-butoxy, and the like.
[0050] "C.sub.2-C.sub.6 alkenyl" refers to those groups such as
vinyl, 1-propene-2-yl, 1-butene-4-yl, 1-pentene-5-yl,
1-butene-1-yl, and the like.
[0051] "C.sub.2-C.sub.10 alkylthio" refers to those groups such as
methylthio, ethylthio, t-butylthio, and the like.
[0052] "C.sub.1-C.sub.4 alkanoyloxy" refers to those groups such as
acetoxy, propionoxy, formyloxy, butyryloxy, and the like.
[0053] The term "C.sub.2-C.sub.8 alkenoxy" includes groups such as
ethenyloxy, propenyloxy, iso-butoxy ethenyl, and the like.
[0054] The term "C.sub.2-C.sub.8 alkynyl" includes groups such as
ethynyl, propynyl, pentynyl, butynyl, and the like.
[0055] The term halo-substituted C.sub.1-C.sub.6 alkyl includes
alkyls substituted 1, 2, or 3 times by a halogen, including groups
such as trifluoromethyl, 2-dichloroethyl, 3,3-difluoropropyl, and
the like.
[0056] The term "amine" includes groups such as NH.sub.2,
NHCH.sub.2 and N(CH).sub.2 and the like which may be optionally
substituted with halogen, amino, C.sub.1-C.sub.7 acyloxy,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, nitro, carboxy,
carbanoyl, carbanoyloxy, cyano or methylsulphonylamino and the
like.
[0057] The terms "carboxyl", "carboxymethyl" and "carbamoyl"
include the corresponding pharmaceutically acceptable
C.sub.1-C.sub.6 alkyl and aryl esters.
[0058] The term "aryl" includes 3 to 8 membered stable saturated or
unsaturated organic monocyclic rings having 0 to 4 hetero atoms
selected from S, O, and N; and 7 to 10 membered organic stable,
saturated or unsaturated, bicyclic rings having 0 to 5 hetero atoms
selected from S, O, N; both of which may be substituted by halo,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, C.sub.2-C.sub.6
alkenyl, C.sub.2-C.sub.6 alkynyl, substituted C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 substituted alkoxy, substituted
C.sub.2-C.sub.6 alkenyl, or substituted C.sub.2-C.sub.6 alkynyl,
substituted alkoxy, amino, nitro, cyano, carboxy, hydroxymethyl,
aminomethyl, carboxymethyl, C.sub.1-C.sub.4 alkylthio, hydroxy,
C.sub.1-C.sub.4 alkanoyloxy, carbamoyl, or halo-substituted
C.sub.1-C.sub.6 alkyl.
[0059] The terms substituted alkyl, substituted alkenyl,
substituted alkynyl and substituted alkoxy are these substitutents
substituted with halogen, hydroxy, amino, C.sub.1-C.sub.7 acyloxy,
nitro, carboxy, carbanoyl, carbanoyloxy, cyano,
methylsulfonylamino, or C.sub.1-C.sub.6 alkoxy, and may be
substituted once or twice with the same or different group.
[0060] Preferred definitions are when R.sub.3 and R.sub.4 are
hydrogen; and
[0061] R.sub.1 is a heteroaromatic monocyclic or bicyclic ring as
defined above and has the N'-linkage at the 3 position, or more
preferably at the 2 position, relative to a hetero atom.
[0062] Examples of R.sub.1 are thiazolyl, (4-methyl)thiazolyl,
(4,5-dimethyl)thiazolyl, (4-cyano)thiazolyl,
(4-trifluoromethyl)thiazolyl- , benzothiazolyl,
(6-fluoro)benzothiazolyl, (6-chloro)pyrazinyl, (4-ethyl)thiazolyl,
4-(3-pyridyl)thiazolyl, 4-(3-nitrophenyl)thiazolyl, pyridyl,
(6-bromo)pyridyl, (6-chloro)pyridyl, (6-methyl)pyridyl,
(5-methyl)pyridyl, (6-trifluoromethyl)pyridyl,
(5-trifluoromethyl)pyridyl- , (6-ethyl)pyridyl, (5-ethyl)pyridyl,
(6-bromo)pyrazinyl, 3-(6-bromo)pyridazinyl, (6-cyano)pyridyl,
(5-cyano)pyridyl, (5-cyano)pyrazinyl, (6-cyano)pyrazinyl,
3-(6-cyano)pyridazinyl, 1,3,4-thiadiazoyl, benzimidazolyl,
imidazolyl, (5-bromo)pyridyl, (5-chloro)pyridyl,
(5-chloro)pyrazinyl, (5-bromo)pyrazinyl, (6-chloro)pyridazinyl,
2-(3-[6-bromo]pyridazinyl), 2-(3-[6-chloro]pyridazinyl),
2-(3-[6-cyano]pyridazinyl), 2-(3-ethyl)pyridyl,
3-(6-methoxy)pyridazinyl, 2-(5-nitro)pyridyl,
2-(5,6-dichloro-4-azabenzimidazolyl), 4-(6-aminopyrimidinyl),
4-pyrimidinyl, 2-(3-pyridazinyl), 2-(3-(6-methyl)pyridazinyl,
2-pyrazinyl, 2-(5-methyl)pyrazinyl); and R.sub.2 are (4-methyl)
-3-pentenyl,
(.+-.)-cis-N-(3,4-benzo-cis-bicyclo-[3.1.0]-hexen-6-yl), 5
[0063] The preferred optional substituents for R.sub.1 comprise
mono, di, or tri halo, preferably bromo or chloro, especially para
to the N'-linkage.
[0064] The preferred optional substituents to R.sub.5 are halo or
C.sub.1-C.sub.6 alkoxy, especially 2,6-difluoro and
2,6-dihalo-3-C.sub.1-C.sub.6 alkoxy.
[0065] Preferred R.sub.5 groups include phenyl, 2-methoxyphenyl,
3-methoxyphenyl, 4-methoxyphenyl, 2-ethoxyphenyl, 2-methylphenyl,
3-methylphenyl, 2-fluorophenyl, 2,6-difluorophenyl,
2-fluoro-6-methoxyphenyl, 2-fluoro-6-ethoxyphenyl-,
2,3,5,6-tetrafluorophenyl, 2-chlorophenyl, 3-chlorbphenyl,
1-cyclohexenyl, 2-naphthyl, 2,5-dimethoxyphenyl, 2-azidophenyl,
2,3,4-trifluorophenyl, 2-fluoro-6-chlorophenyl,
2,6-dimethoxyphenyl, 2,3,6-trichlorophenyl, 2,6-dichlorophenyl,
2,3,5-trichlorophenyl, 3,5-dichlorophenyl, 3-fluorophenyl,
2,4-dimethoxyphenyl, 2-pyridyl, 2-(6-methoxy)pyridyl,
2-(6-ethoxy)pyridyl, 2-(6-fluoro)pyridyl, 2-(5-fluoro)pyridyl,
2-(4-fluoro)pyridyl, 2-(3-fluoro)pyridyl, 2-(6-chloro)pyridyl,
2-(5-chloro)pyridyl, 2-(4-chloro)pyridyl, 2-(3-chloro)pyridyl,
2-(5-methoxy-6-fluoro)pyridyl, 2-(3-methoxy-6-fluoro)pyridyl,
2-(6-methoxy-3-fluoro)pyridyl, 2-(5-ethoxy-6-fluoro)pyridyl,
2-(3-ethoxy-6-fluoro)pyridyl, 2-(6-ethoxy-3-fluoro)pyridyl,
2-(5,6-difluoro)-pyridyl, 2-(3,6-difluoro)pyridyl,
2-(5,6-dichloro)-pyridyl, 2-(3,6-dichloro)pyridyl,
2-(6-methoxy)pyridyl, 2-(6-ethoxy)pyridyl, 2-[1,3-pyrimidyl],
2-pyrazinyl, 3-pyridazinyl, 2,6-difluoro-3-methoxyphen- yl,
2,6-difluoro-3-ethoxyphenyl, 2,6-difluoro-4-methoxyphenyl,
2,6-difluoro-4-ethoxyphenyl, 2-(3-ethoxy)pyridyl,
2-(3-methoxy)pyridyl, 2,6-difluorophenyl,
2,6-difluoro-3-N-methyl-carboxamidephenyl, 2-fluoro-6-chlorophenyl,
3-bromo-6-methoxyphenyl, 3-ethoxyphenyl, 3-bromo-6-ethoxyphenyl,
3-(2-fluoro)pyridyl, (2-vinyl)phenyl, (3-vinyl)phenyl,
(3-methoxy-carbonyl)phenyl, 5,6-dimethylbenzotriazolyl,
2,3-difluoro-6-methoxyphenyl, 2,6-difluoro-3-cyanophenyl,
3-ethynylphenyl, and 2,5-diethoxyphenyl.
[0066] A further aspect of the invention provides novel compounds
within the above Formula 1A, wherein A is as defined above;
[0067] R.sub.1 is a stable, unsaturated, substituted or
unsubstituted heterocycle having i) a 3 to 8 membered monocyclic
ring with up to 4 hetero atoms or ii) a 7 to 10 membered bicyclic
ring with up to 5 hetero atoms wherein the N'-bonded ring has at
least one of the hetero atoms and wherein the hetero atoms are
selected from N, O and S;
[0068] R.sub.2 is a group of the formula 6
[0069] wherein R.sub.5 is a stable, saturated or unsaturated,
substituted or unsubstituted, i) 3 to 8 membered organic monocyclic
ring having 0 to 4 hetero atoms or ii) a 7 to 10 membered organic
bicyclic ring having 0 to 5 hetero atoms, the hetero atoms being
selected from S, O and N;
[0070] and R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are independently
C.sub.3-C.sub.8 cycloalkyl, hydrogen, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, substituted
C.sub.1-C.sub.6 alkyl, substituted C.sub.2-C.sub.6 alkenyl,or
substituted C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 substituted
alkoxy, halo, amino, nitro, cyano, C.sub.1-C.sub.5 alkoxy, hydroxy,
hydroxymethyl, aminomethyl, carboxymethyl, C.sub.1-C.sub.4
alkylthio, C.sub.1-C.sub.4 alkanoyloxy, carbamoyl, or a halo
substituted C.sub.1-C.sub.6 alkyl; or two of which, along with the
carbons to which they are attached, combine to form a stable,
saturated or unsaturated, substituted or unsubstituted, 3 to 7
membered organic monocylic ring having 0 to 4 hetero atoms selected
from S, O, or N; or R.sub.6 and R.sub.8, or R.sub.7 and R.sub.9,
along with the carbon to which they are attached, form a stable,
saturated or unsaturated, substituted or unsubstituted, 3 to 7
membered organic monocylic ring having 0 to 4 hetero atoms selected
from S, O, or N; or a pharmaceutically acceptable salt thereof,
with the provisos that:
[0071] a) if R.sub.6-R.sub.9 are all hydrogen, then the following
combinations are excluded:
1 A R.sub.1 R.sub.5 i O 6-chloropyridazin-3-yl morpholine ii O
1-oxy,2,2,5,5 tetramethyl- isoindole pyrrol-3-yl iii O
(substituted) 4-carboxy- piperazine, fur-3-yl (sub- stituted)
phenyl iv O benzimidazol-2-yl phenyl v O purin-6-yl imidazole,
(sub- stituted) phenyl vi O (substituted) oxazol-2-yl phenyl vii O
benzothien-3-yl morpholine viii NH pyrimidin-2-yl phenyl ix NH
imidazol-2-yl (sub- stituted) phenyl x NCN pyrid-3-yl phenyl
[0072] b) if R.sub.7 is methyl or together with R.sub.6 forms
cyclopropyl and the remainder of R.sub.6-R.sub.9, are hydrogen,
then the following combination is excluded:
2 A R.sub.1 R.sub.5 O pyrid-2-yl phenyl; and
[0073] c) if R.sub.7 is alkylthio, and the remainder of
R.sub.6-R.sub.9 are hydrogen, then the following combination is
excluded
3 A R.sub.1 R.sub.5 O pyrid-3-yl phenyl.
[0074] The definitions above also apply to these compound aspects
of the invention with the exception that aryl and substituted aryl
are excluded as possible optional substituents to group
R.sub.1.
[0075] Further, preferred compounds of the invention are those in
which R.sub.1, and R.sub.6-R.sub.9 are as defined above and R.sub.5
is substituted or unsubstituted C.sub.2-C.sub.6 alkenyl,
C.sub.2-C.sub.6 alkynyl or wherein R.sub.5 is --OR.sub.10 or
--C(O)R.sub.10 where R.sub.10 is R.sub.5 as defined above, or
substituted or unsubstituted C.sub.2-C.sub.6 alkenyl or
C.sub.2-C.sub.6 alkynyl.
[0076] It is believed that none of, the compounds excluded by the
provisos except proviso x) have been known in a therapeutic role
and thus, a further aspect of the invention includes the compounds
embraced by the provisos for use in therapy.
[0077] As mentioned above, the invention includes pharmaceutically
acceptable salts of the compounds defined by the above formula (I).
Although generally neutral, a particular compound of this invention
can possess a sufficiently acidic, a sufficiently basic, or both,
functional groups, and accordingly react with any of a number of
nontoxic inorganic bases, and nontoxic inorganic and organic acids,
to form a pharmaceutically acceptable salt. Acids commonly employed
to form acid addition salts are inorganic acids such as
hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric
acid, phosphoric acid and the like, and organic acids such as
p-toluene sulfonic, methanesulfonic acid, oxalic acid,
p-bromophenylsulfonic acid, carbonic acid, succinic acid, citric
acid, benzoic acid, acetic acid, and the like. Examples of such
pharmaceutically acceptable salts thus are the sulfate,
pyrosulfate, bisulfate, sulfite, bisulfite, phosphate,
monohydrogenphosphate, dihydrogenphosphate, metaphosphate,
pyrophosphate, chloride, bromide, iodide, acetate, propionate,
decanoate, caprylate, acrylate, formate, isobutyrate, caproate,
heptanoate, propiolate, oxalate, malonate, succinate, suberate,
sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate,
benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate,
hydroxybenzoate, methoxybenzoate, phthalate, sulfonate,
xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate,
citrate, lactate, g-hydroxybutyrate, glycollate, tartrate,
methanesulfonate, propanesulfonate, naphthalene-1-sulfonate,
naphthalene-2-sulfonate, mandelate and the like. Preferred
pharmaceutically acceptable acid addition salts are those formed
with mineral acids such as hydrochloric acid and hydrobromic acid,
and those formed with organic acids such as maleic acid and
methanesulfonic acid.
[0078] Base addition salts include those derived from inorganic
bases, such as ammonium or alkali or alkaline earth metal
hydroxides, carbonates, bicarbonates, and the like. Such bases
useful in preparing the salts of this invention thus include sodium
hydroxide, potassium hydroxide, ammonium hydroxide, potassium
bicarbonate, calcium hydroxide, calcium carbonate, and the
like.
[0079] The pharmaceutically acceptable salts of the invention are
typically formed by reacting a compound as defined with an
equimolar or excess amount of acid or base. The reactants are
generally combined in a mutual solvent such as diethyl ether or
benzene, for acid addition salts, or water or alcohols for base
addition salts, and the salts normally precipitate out of solution
within about one hour to about ten days and can be isolated by
filtration or other conventional methods. The salts of the
compounds of the invention will convert to the compound per se
after administration and are thus prodrugs. All prodrugs are
administered in an amount sufficient to generate an effective
amount of the compound to contact the virus and interact with it
(e.g. inhibit replication thereof).
[0080] The compounds of the present invention also include
racemates, racemic mixtures, and individual enantiomers or
diastereomers. All asymmetric forms, individual isomers and
combinations thereof are within the scope of the present
invention.
[0081] As noted, the optically active diastereomers of the
compounds of Formula 1 are considered part of this invention and
such optically active isomers may be prepared from their respective
optically active precursors by the procedures described herein, or
by resolving the racemic mixtures. The resolution can be carried
out in the presence of a resolving agent, by chromatography, by
repeated crystallization or by some combination of these techniques
which are known to those skilled in the art. Further details
regarding resolutions can be found in Jacques, et al., Enantiomers,
Racemates, and Resolutions, John Wiley & Sons 1981.
[0082] The compounds of the present invention, or their precursors,
are prepared using procedures known to those of ordinary skill in
the art. More particularly, the compounds of Formula (1) are
prepared according to the general procedures shown below in Schemes
I-IV, and VII, as described below.
[0083] In Scheme I, a derivative of an isocyanate 7
[0084] or isoselenocyanate 8
[0085] is reacted with with an amino group (2) in approximately 1:1
molar ratio, in an inert organic solvent, such as N,N-dimethyl
formamide and stirred at an appropriate temperature of between
about 0-150.degree. C. for a period of, time between 1 and 72
hours. The time and temperature used depends upon the reactivity of
the individual reagents. The product (3) may be isolated by
conventional techniques.
[0086] Scheme II is run under the same general reaction conditions
as Scheme I. Where 9
[0087] Schemes I and II are processes analogous to that described
in Advanced Org. Chem. 3rd Ed., J. March, p 802, 1985, J. Wiley
& Sons and references cited therein, incorporated herein by
reference. Wherein 10
[0088] Schemes I and II are processes analogous to that described
in Ber. Deutsch Chem. Ges., 100, 1459, 1967; J. Chem. Soc. Chem.
Commun., 372, 1968; J. Prakt. Chem., 315, 155, 1973 and references
cited therein, incorporated herein by reference. Wherein
A=SO.sub.2, Schemes I and II are processes analogous to that
described in J. Am. Chem. Soc., 94, 6135, 1972 and Org. Prep.
Proced. Int., 16, 49, 1984 and references cited therein,
incorporated herein by reference. 11 12
[0089] In Scheme III, a derivative of an amine (7) and an amine (8)
are added in an inert solvent, such as N,N-dimethyl formamide,
tetrahydrofuran, ethyl acetate, or benzene, to an appropriately
substituted derivative (9) wherein L.sub.1 and L.sub.2 represent
leaving groups. The reaction is stirred at an appropriate
temperature of between about 0-150.degree. C. for a period of time
between 1 and 72 hours. The time, temperature, and sequence of
addition used depends upon the reactivity of the individual
reagents. The product (10) may be isolated by conventional
techniques.
[0090] Wherein 13
[0091] Scheme III is a process analogous to that described in J.
Org. Chem., 56, 891, 1991 and references cited therein,
incorporated herein by reference.
[0092] Wherein 14
[0093] Scheme III is a process analogous to that described in Bull.
Chem. Soc. Jpn., 62, 2419, 1989 and references cited therein,
incorporated herein by reference.
[0094] Wherein 15
[0095] Scheme III is a process analogous to that described in J.
Org. Chem., 24 , 1157, 1959; J. Org. Chem., 38, 155, 1973; J. Med.
Chem., 20, 901, 1977; Tetrahedron Lett., 30, 7313, 1989; Orq. Prep.
Proced. Int., 17, 256, 1985; J. Heterocyclic Chem., 24, 275, 1987;
Org. Prep. Proced. Int., 23, 721, 1991 and references cited
therein, incorporated herein by reference.
[0096] Wherein 16
[0097] Scheme III is a process analogous to that described in
Angew. Chem. Int. Ed., Enq., 26, 1165, 1987; Liebigs Ann. Chem.,
207, 1990; European Patent 58492, 1982; Syn. Commun., 19, 943,
1989; J. Heterocyclic Chem., 26, 1335, 1989; Belgium Patent 894093,
1982; Syn. Commun., 21, 1213, 1991; Synthesis, 195, 1990; U.S. Pat.
No. 4,968,808; J. Org. Chem., 32, 2661, 1967; Synthesis, 76, 1987
and references cited therein, incorporated herein by reference.
Wherein A=SO.sub.2, Scheme III is a process analogous to that
described in Synthesis, 192, 1983; J. Org. Chem., 45, 5371, 5373,
1980; Org. Syn. Coll. Vol. VI, 78, 1988; J. Prakt. Chem., 29, 328,
1965; Arch. Pharm., 321, 375, 1988; J. Chem. Soc., 4367, 1960;
Synthesis, 576, 1972 and references cited therein, incorporated
herein by reference. 17
[0098] In Scheme IV, a thiourea derivative (11) is converted to the
corresponding derivative (10). Wherein 18
[0099] Scheme IV is a process analogous to that described in J.
Org. Chem., 26, 2925, 1961 and Tetrahedron Lett., 27, 3911, 1986
and references cited therein, incorporated herein by reference.
Wherein 19
[0100] Scheme IV is a process analogous to that described in Eur.
J. Med. Chem., 16, 317, 1981 and references cited therein,
incorporated herein by reference. Wherein 20
[0101] Scheme IV is a process analogous to that described in J.
Org. Chem., 49, 4123, 1984; J. Med. Chem., 20, 901, 1977;
Tetrahedron Lett., 30, 7313, 1989; Syn. Commun., 13, 67, 1983; and
Curr. Sci., 37, 645, 1968 and references cited therein,
incorporated herein by reference. 21
[0102] In Scheme V, a carbodiimide derivative (12) is reacted with
a nucleophile to form the corresponding product (13). Wherein
22
[0103] Scheme V is a process analogous to that described in U.S.
Pat. No. 4,414,211, 1980; J. Chem. Soc. C, 1429, 1970; J. Chem.
Soc. P. T. 1, 1241, 1977; J. Med. Chem., 32, 228, 1989 and
references cited therein, incorporated herein by reference. Wherein
23
[0104] Scheme V is a process analogous to that described in Monat.
fur Chem., 97, 695, 1966 and Chem. Ztq., 112, 107, 1988 and
references cited therein, incorporated herein by reference. 24
[0105] In Schemes VI and VII, derivatives 14 and 15, wherein L
represents a leaving group, is reacted with a nucleophile to afford
the corresponding derivatives 16 and 17, respectively. Wherein
25
[0106] Schemes VI and VII are processes analogous to that described
in Orq. Syn. Coll. Vol. V, 966, 1973 and references cited therein,
incorporated herein by reference. Wherein 26
[0107] Schemes VI and VII are processes analogous to that described
in J. Org. Chem., 51, 1882, 1986; Syn. Comm., 20, 217, 1990,
Tetrahedron Lett., 1879, 1970, Org. Prep. Proced. Int., 17, 256,
1985; J. Heterocyclic Chem., 24, 275, 1987; Org. Prep. Proced.
Int., 23, 721, 1991 and references cited therein, incorporated
herein by reference. Wherein 27
[0108] Scheme VI and Vll is a process analogous to that described
in Ann. Chem., 2096, 1979 and references cited therein,
incorporated herein by reference. 28 29
[0109] Compounds wherein Z.sub.ii is S(.dbd.0).sub.2 can be
prepared by reacting an amine of the formula R.sub.2NH.sub.2 with
diaminosulfone, for instance under reflux in 1,2-dimethoxymethane,
to form a compound of the formula R.sub.2NHS(.dbd.0).sub.2NH.sub.2.
This, in turn, is subjected to reaction with a compound of the
formula H.sub.2NR.sub.1, for instance, in DMF or
1,2-dimethoxyethane, to form R.sub.2NHS(.dbd.0).sub.2NHR.sub.1.
This process is, thus, analogous to that described in Arya et al.,
Ind. J. Chem. B, 1976, 14B, 766.
[0110] Alternatively, compounds wherein Z.sub.ii is sulphone can be
prepared by reacting an amine of the formula R.sub.2NH.sub.2 with a
dihalosulphone, e.g. SO.sub.2Cl.sub.2, for instance under reflux
with SbCl.sub.5 and acetonitrile to produce a compound of the
formula R.sub.2NHS(.dbd.0).sub.2halo which can be reacted with the
appropriate R.sub.1 amine, for example 2-amino, 6-chloropyridyl in
acetonitrile and triethylamine. This process is analogous to
Liebigs Ann. Chem. 729 (1969) 40.
[0111] Compounds wherein Z.sub.ii is SO can be prepared by reducing
compounds prepared by the above two methods using conventional
reduction techniques. Alternatively, a sulfinyl compound can be
prepared by the methodology in Liebigs Ann. Chem. 1979, 1756, where
SOCl.sub.2 is reacted with several molar equivalents of an aryl
leaving group such as pyridyl to form a dipyrid-1-ylsulphinyl
intermediate. This, in turn, can be transaminated with appropriate
R.sub.1 and R.sub.2 amines as described in Tetrah. Lett. 1985, 26,
3821, to produce a compound of the structure
R.sub.2NHSONHR.sub.1.
[0112] Tests with the above compounds of Formula 1 have indicated
activity as inhibitors of HIV. While not being bound by theory, it
is believed that the compounds act as reverse transcriptase
inhibitors, and thereby act to inhibit replication of the
virus.
[0113] The HIV inhibition activity of the present invention is
generally between 1 and 3 logs greater against HIV-1 strains than
against HIV-2 strains. This enables the compounds and compositions
of the present invention to be used to selectively inhibit HIV-1
growth in cell cultures where co-infection with HIV-1 and HIV-2 is
suspected. Suppression of one strain or the other is necessary to
enable appropriate choice of antiviral agent to combat each strain.
In practice, selective inhibition of HIV-1 in a co-infected cell
culture entails administering a concentration of a compound of the
present invention which is intermediate the respective IC.sub.50 or
more preferably the IC.sub.90 for that compound against HIV-1 and
HIV-2. The cell culture is subject to an appropriate incubation
time, which is selected to be sufficiently long to disable the
infectivity of the HIV-1 virions, but not sufficiently long as to
encourage the development of resistant mutants, for example between
24 and 48 hours. The supernatent is subject to limiting dilution,
preferably in a conventional resuscitation buffer to sequester any
remaining compound, until it is statistically likely that only a
single infectious dose of HIV-2 virons is left. This is then
reinoculated in order to determine, for example, the resistance
profile or anti-viral susceptibility of the HIV-2 strain
uninfluenced by the co-infection with HIV-1.
[0114] The following is a description of the test systems used in
analyzing compounds in effectiveness against HIV.
[0115] Tests A, B, C, and D (XTT)
[0116] MT4 cells in a medium of RPMI 1640, 5% FCS,
penicillin/streptomycin are adjusted to 2.times.10.sup.5 cells/ml
and seeded into microplates (96 wells/plate) 100 ml cell
suspension/well giving 2.times.10.sup.4 cells/well. The compound to
be tested is made into a 10 mg/ml mixture in DMSO and stored at
-20.degree. C. The compound in DMSO is diluted with medium
containing 10% DMSO in a 10-fold dilution series to give 1 mg/ml,
10 mg/ml, and 100 mg/ml solutions. Further dilutions to 400, 40, 4
and 0.4 mg/ml are made in medium containing microplates. Fifty ml
of the 400, 40, and 4 mg/ml are transferred to the cell-containing
microplates with a multi-channel pipette (final concentration: 100,
10, and 1 mg/ml). Finally, 50 ml of virus suspension is added to
each well (with a repetitive "Eppendorf" multipipett). Each plate
has at least six wells with the following: [Test A: HIV virus; Test
B: HIV(II) virus; Test C: SIV virus; Test D: No virus]; with no
drug (virus control) and six wells without virus (medium control).
The plate is put into a plastic bag and incubated for six days in
CO.sub.2 atmosphere.
[0117] To each well in the plate is added 50 ml of XTT
((2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-5-[(phenylamin
o)carbonyl]-2H-tetrazolium hydroxide), (1 mg/ml 0.01-0.02 mM
N-metyl-phenazonium methosulfate). After six hours of incubation in
C.sub.0.sub.2 atmosphere the plates are covered with adhesive plate
sealers and gently mixed on a vortex. Optical densities are
determined at a wavelength of 450 nm and a reference wavelength of
650 nm. The percent reduction of cytotoxocity caused by the virus
infection is calculated as follows: 1 OD 450 compound - OD 450 inf
cells OD 450 uninf cells - OD 450 inf cells .times. 100
[0118] Tests E, F, G, H (HIV-IRT, HIV-2RT, SIVRT, no virus)
[0119] MT-4/H9-cells are adjusted to 2.times.10.sup.5 cels/ml
medium (RPMI 1640, 5% FCS, penicillin/streptomycin) and seeded into
microplates (96 wells/plate) 100 ml cell suspension/well giving
2.times.10.sup.4 cells/well. The compound to be tested is made 10
mg/ml in DMSO=stock solution (stored at -20.degree. C.). The
compound dissolved in DMSO is diluted 25 times in medium to give
400 mg/ml. Further dilutions to 40 mg/ml and 4 mg/ml are made in
microplates.
[0120] 50 ml of the dilutions 400 mg/ml, 40 mg/ml and 4mg/ml are
transferred to the "cell-containing" microplate with a multichannel
pipette. (Final concentration: 100, 10 and 1 mg/ml).
[0121] Finally 50 ml of virus suspension is added to each well
(with a repetitive "Eppendorf multipett"). [Test E-HIV-1; Test
F-HIV-2; Test G-SIV; Test H-no virus].
[0122] Each plate has at least four wells with virus but no drug
(virus control) and two wells without virus (medium control). The
plate is put into a plastic bag to avoid evaporation and incubated
for six days in CO.sub.2-atmosphere.
[0123] 10 ml supernatant from each well is transferred with a
multichannel pipette into a new microplate to which 40 ml VDB, (50
mM Tris-HCl pH=7.6, 35 mM KCl, 4 mM DTT, 1 mM EDTA, 1.3% Triton
X-100), have been added to each well. The addition of 50 ml
RT-reaction mix, (10 ml culture supernatant, 40 ml VDB and 50 ml
reaction mixture giving a final concentration of: 100 mM Tris-HCl
pH=7.6, 100 mM KCl, 4 mM MgCl.sub.2, 4 mM DTT, 275 mg/ml BSA/ml, 5
mg (rA).sub.n(dT).sub.12-18/ml and 0.3 mM .sup.3H dTTP (specific
activity 18.000 cpm/pmol)) gives a final volume of 100 ml/well.
After 60 minutes of incubation the whole assay volume is
transferred by use of a cell harvester to a filter mat prewetted
with 5% TCA. The filter is washed in 5% TCA and rinsed once in
ethanol. After drying the filter mat at 60.infin. C. for 30 min.
each filter (96/mat) is punched out and put into counting vials 2
ml of scintillation fluid is added and the samples are counted (1
min) or the whole filter mat is put into a plastic bag, 10 ml of
scintillation fluid is added and the filter mat is counted in a
Beckman Betaplate counter. Percent reduction of RT activity is
determined by comparing RT activity for virus control with the RT
activity measured for each dilution of the compound.
[0124] Test I (HIVRT (rAdt))
[0125] The compounds were tested for direct inhibitory activity on
HIV-RT in a volume of 100 ml recombinant HIV-RT (diluted in virus
disruption buffer to give 200.000 cpm).
[0126] 100 mM Tris-HCl pH 7.6, 100 mM KCl, 4 mM MgCl.sub.2, 4 mM
DTT, 275 mg/ml BSA, 0.5 mg (rA)n(dT).sub.12-18 and 0.3 mM
.sup.3H-=dTTP (specific activity 18.000 cpm/mol). After 60 minutess
of incubation 40 ml in duplicate were spotted on paper discs and
washed in 5% TCA. After rinsing the paper discs in ethanol they
were dried and counted in scintillation fluid.
[0127] The following illustrate activities of compounds in the
above-described tests. The numbers represent % inhibition. 30
4 100 10 1 0.1 0.01 0.001 0.1 IC50 Test .mu.g/ml .mu.g/ml .mu.g/ml
.mu.g/ml .mu.g/ml .mu.g/ml .mu.g/ml .mu.g/ml I 100 98 89 55 6
0.0008 I 100 97 93 62 16 0.0005 A 3 100 86 82 80 7 A 100 100 90 100
81 8
[0128] 31
5 100 10 1 0.1 0.01 0.001 0.1 IC50 Test .mu.g/ml .mu.g/ml .mu.g/ml
.mu.g/ml .mu.g/ml .mu.g/ml .mu.g/ml .mu.g/ml I 98 96 82 41 13 0.002
I 100 100 83 45 18 0.0014 A 100 100 98 92 96 5 A 100 83 99 89 82
47
[0129] 32
6 100 10 1 0.1 0.01 0.001 0.1 IC50 Test .mu.g/ml .mu.g/ml .mu.g/ml
.mu.g/ml .mu.g/ml .mu.g/ml .mu.g/ml .mu.g/ml I 99 96 82 46 19
0.0011 I 97 96 80 30 5 0.002 A 24 88 87 100 100 0 A 30 100 100 100
100 19
[0130] 33
7 100 10 1 0.1 0.01 0.001 0.1 IC50 Test .mu.g/ml .mu.g/ml .mu.g/ml
.mu.g/ml .mu.g/ml .mu.g/ml .mu.g/ml .mu.g/ml I 100 100 91 58 17
0.0006 A 0 100 87 92 51 0 A 0 93 98 100 94 6
[0131]
8 Example Test IC.sub.50 (.mu.g/ml) 56 I 0.0023, 0.0022 57 I
0.0004, 0.0005, 0.0004 58 I 0.038 26 I 0.009, 0.01 27 I 0.0005,
0.0006 23 I 0.005, 0.014 34 I 0.0004 33 I 0.0005 21 I 0.05 64 I
0.0013
[0132] A feature of this invention also disclosed is a method of
administering to a human in need thereof the compounds described,
their pharmaceutically acceptable salts or pro-drugs thereof to
treat or inhibit HIV/AIDS, to inhibit the replication of the
HIV/AIDS virus in infected human cells and to inhibit AIDS from
developing n humans infected with the HIV/AIDS virus or carrying
antibodies to the HIV/AIDS virus.
[0133] The present invention also discloses the compounds of the
invention and their salts for use in the treatment of the condition
referred to above, as well as the use of such compounds in the
preparation of pharmaceutical formulations for the treatment of
such conditions.
[0134] In general for the treatment as described above, a suitable
effective dose of the compound or its pharmaceutically acceptable
salt will be in the range of 0.5 to 250 mg per kilogram bodyweight
of recipient per day. Administration may be by any suitable route
including oral, rectal, nasal, topical (including buccal and
sublingual), vaginal or parenteral (including subcutaneous,
intramuscular, intravenous and intradermal) administration. It will
be appreciated that the preferred route may vary with, for example,
the condition, age, and weight of the recipient.
[0135] The administered ingredients may be used as a therapy in
conjunction with other therapeutic agents, (other anti-virals,
anti-bacterials, compounds useful for preventing resulting
secondary or contemporaneous afflictions associated with HIV/AIDS)
such as AZT, ddI, ddC,
9-[[2-hydroxy-1-(hydroxymethyl)-ethoxy]methyl]guanine,
9-(2-hydroxyethoxymethyl)-guanine(acyclovir),
2-amino-9-(2-hydroxyethoxym- ethyl)-purine, suramin, ribavarin,
antimoniotungstate (HPA-23), interferon, e.g., a interferon,
interleukin II, and phosphonoformate (Foscarnet) or in conjunction
with other immune modulators including bone marrow or lymphocyte
transplants or other medications such as levamisol or thymosin
which would increase lymphocyte numbers and/or function as is
appropriate.
[0136] While it is possible for the administered ingredients to be
administered alone, it is preferable to present them as part of a
pharmaceutical formulation. The formulations of the present
invention comprise at least one administered ingredient, as
above-defined together with one or more acceptable carriers thereof
and optionally other therapeutic ingredients. The carrier(s) must
be "acceptable" in the sense of being compatible with the other
ingredients of the formulation and not deleterious to the recipient
thereof.
[0137] The formulations include those suitable for oral, rectal,
nasal, topical (including buccal and sublingual), vaginal or
parenteral (including subcutaneous, intramuscular, intravenous and
intradermal) administration. The formulations may conveniently be
presented in unit dosage form, e.g., tablets and sustained release
capsules, and may be prepared by any methods well known in the art
of pharmacy.
[0138] Such methods include the step of bringing into association
the to be administered ingredients with the carrier which
constitutes one or more accessory ingredients. In general, the
formulations are prepared by uniformly and intimately bringing into
association the active ingredients with liquid carriers or finely
divided solid carriers or both, and then if necessary shaping the
product.
[0139] Formulations of the present invention suitable for oral
administration may be presented as discrete units such as capsules,
cachets or tablets each containing a predetermined amount of the
active ingredient; as a powder or granules; as a solution or a
suspension in an aqueous liquid or a non-aqueous liquid; or as an
oil-in-water liquid emulsion or a water-in-oil liquid emulsion and
as a bolus, etc.
[0140] With regard to compositions for oral administration (e.g.
tablets and capsules), the term "suitable vehicle" means common
excipients such as binding agents, for example, syrup, acacia,
gelatin, sorbitol, tragacanth, polyvinylpyrrolidine (Povidone),
methylcellulose, ethylcellulose, sodium carboxy-methylcellulose,
hydroxypropylmethylcellul- ose, sucrose and starch; fillers and
carriers, for example corn starch, gelatin, lactose, sucrose,
microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate,
sodium chloride and alginic acid; disintegrators such as
microcrystalline cellulose, corn starch, sodium starch glycolate,
alginic acid; and lubricants such as magnesium stearate and other
metallic stearates, stearic acid, silicone fluid, talc, waxes, oils
and colloidal silica. Flavoring agents such as peppermint, oil of
wintergreen, cherry flavoring or the like can also be used. It may
be desirable to add a coloring agent to make the dosage form more
aesthetically pleasing in appearance or to help identify the
product. The tablets may also be coated by methods well known in
the art.
[0141] A tablet may be made by compression or molding, optionally
with one or more accessory ingredients. Compressed tablets may be
prepared by compressing in a suitable machine the active ingredient
in a free-flowing form such as a powder or granules, optionally
mixed with a binder, lubricant, inert diluent, preservative,
surface-active or dispersing agent. Molded tablets may be made by
molding in a suitable machine a mixture of the powdered compound
moistened with an inert liquid diluent. The tablets may optionally
be coated or scored and may be formulated so as to provide slow or
controlled release of the active ingredient therein.
[0142] Formulations suitable for topical administration include
lozenges comprising the ingredients in a flavored basis, usually
sucrose and acacia or tragacanth; pastilles comprising the active
ingredient in an inert basis such as gelatin and glycerin, or
sucrose and acacia; and mouthwashes comprising the ingredient to be
administered in a suitable liquid carrier.
[0143] Formulations suitable for topical administration to the skin
may be presented as ointments, creams, gels and pastes comprising
the ingredient to be administered and a pharmaceutically acceptable
carrier. An exemplary topical delivery system is a transdermal
patch containing the ingredient to be administered.
[0144] Formulations for rectal administration may be presented as a
suppository with a suitable base comprising, for example, cocoa
butter or a salicylate.
[0145] Formulations suitable for nasal administration wherein the
carrier is a solid include a coarse powder having a particle size,
for example, in the range 20 to 500 microns which is administered
in the manner in which snuff is taken, i.e., by rapid inhalation
through the nasal passage from a container of the powder held close
up to the nose. Suitable formulations wherein the carrier is a
liquid, for administration, as for example, a nasal spray or as
nasal drops, include aqueous or oily solutions of the active
ingredient.
[0146] Formulations suitable for vaginal administration may be
presented as pessaries, tampons, creams, gels, pastes, foams, or
spray formulations containing in addition to the active ingredient
such carriers as are known in the art to be appropriate.
[0147] Formulations suitable for parenteral administration include
aqueous and non-aqueous sterile injection solutions which may
contain anti-oxidants, buffers, bacteriostats and solutes which
render the formulation isotonic with the blood of the intended
recipient; and aqueous and non-aqueous sterile suspensions which
may include suspending agents and thickening agents. The
formulations may be presented in unit-dose or multi-dose
containers, for example, sealed ampules and vials, and may be
stored in a freeze-dried (lyophilized) condition requiring only the
addition of the sterile liquid carrier, or example water for
injections, immediately prior to use. Extemporaneous injection
solutions and suspensions may be prepared from sterile powders,
granules, and tablets of the kind previously described.
[0148] Preferred unit dosage formulations are those containing a
daily dose or unit, daily sub-dose, as herein above recited, or an
appropriate fraction thereof, of the administered ingredient.
[0149] The antiviral compounds of Formula I can be used as surface
disinfectants. Solutions containing as little as 0.1 percent by
weight of the antiviral compound maybe effective for disinfecting
purposes. Preferably, such solutions also can contain a detergent
or other cleansing agent. The solutions maybe useful for
disinfecting objects such as glassware, dental and surgical
instruments, and surfaces such as walls, floors, and tables in
areas where maintenance of sterile conditions is important, for
example, hospitals, food-preparation areas, and the like.
[0150] In practicing the method for treating or inhibiting HIV
and/or AIDS, the antiviral can be administered in a single daily
dose or in multiple doses per day. The treatment regime may require
administration over extended periods of time, e.g., for several
days or for several months or years. The amount administered per
dose or the total amount administered will depend on such factors
as the nature and severity of the infection, the age and general
health of the patient, the tolerance of both the patient and the
microorganism or microorganisms involved in the infection to the
antiviral compound.
[0151] The following formulation examples represent specific
pharmaceutical formulations employing compounds comprehended by the
present method. The formulations may employ as active compounds any
of the compounds of Formula I or a pharmaceutically acceptable salt
thereof.
[0152] The examples are illustrative only and are not intended to
limit the scope of the invention in any way.
Formulation 1
[0153] Hard gelatin capsules are prepared using the following
ingredients:
9 Quantity (mg/capsule) Compound 1250 Starch dried 200 Magnesium
stearate 10
[0154] The above ingredients are mixed and filled into hard gelatin
capsules in 460 mg quantities.
Formulation 2
[0155] A tablet formula is prepared using the ingredients
below:
10 Quantity (mg/tablet) Compound 250 Cellulose, microcrystalline
400 Silicon dioxide, fumed 10 Stearic acid 5 Magnesium stearate
10
[0156] The components are blended and compressed to form tablets
each weighing 675 mg.
Formulation 3
[0157] An aerosol solution is prepared containing the following
components:
11 Weight Compound 0.25 Ethanol 29.75 Propellant 22 70.00
(Chlorodifluoromethane)
[0158] The active compound is mixed with ethanol and the mixture
added to a portion of the propellant 22, cooled to -30.degree. C.
and transferred to a filling device. The required amount is then
placed in a stainless steel container and diluted with the
remainder of the propellant. The valve units are then fitted to the
container.
Formulation 4
[0159] Tablets each containing 60 mg of active ingredient are made
up as follows:
12 Compound 60 mg Starch 45 mg Microcrystalline cellulose 35 mg
Polyvinylpyrrolidone 4 mg (as 10% solution in water) Sodium
carboxymethyl starch 4.5 mg Magnesium stearate 0.5 mg Talc 1 mg
[0160] The active ingredient, starch and cellulose are passed
through a No. 45 mesh U.S. sieve and mixed thoroughly. The solution
of polyvinylpyrrolidone is mixed with the resultant powders which
are then passed through a No. 14 mesh U.S. sieve. The granules so
produced are dried at 40.degree.-60.degree. C. and passed through a
No. 18 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium
stearate and talc, previously passed through a No. 60 mesh U.S.
sieve, are then added to the granules which, after mixing, are
compressed on a tablet machine to yield tablets each weighing 150
mg.
Formulation 5
[0161] Capsules each containing 80 mg of medicament are made as
follows:
13 Compound 80 mg Starch 59 mg Microcrystalline cellulose 59 mg
Silicone fluid 2 mg
[0162] The active ingredient, cellulose, starch and magnesium
stearate are blended, passed through a No. 45 mesh U.S. sieve, and
filled into hard gelatin capsules in 200 mg quantities.
Formulation 6
[0163] Suppositories each containing 225 mg of medicament are made
as follows:
14 Compound 225 mg Saturated fatty acid 2 mg glycerides
[0164] The active ingredient is passed through a No. 60 mesh U.S.
sieve and suspended in the saturated fatty acid glycerides
previously melted using the minimum heat necessary. The mixture is
then poured into a suppository mold of nominal 2 g capacity and
allowed to cool.
Formulation 7
[0165] An intravenous formulation is prepared as follows:
15 Compound 100 mg Isotonic saline 1000 ml
[0166] The solution of the above ingredients is administered
intravenously at a rate of 1 ml/minute to a mammal in need of
treatment.
[0167] It should be understood that in addition to the ingredients
particularly mentioned above the formulations of this invention may
include other agents conventional in the art having regard to the
type of formulation in question.
[0168] The compounds of the present invention are also useful as
laboratory tools for monitoring mutation of reverse transcriptase
in vitro. For example, the compounds of Formula 1A stabilize the
reverse transcriptase enzyme in vitro and allow for convenient
cocrystallization. In its native state, reverse transcriptase is
difficult to crystallize, a necessary initial step for X-ray
crystallographic determination of the spatial structure of the
enzyme. Repeated monitoring of the fine structure of this enzyme is
necessary due to its rapid mutation in vivo, which in turn, leads
to rapid viral resistance against conventional anti-HIV agents such
as AZT and ddI. The rapid determination of the mutated structures
is a key step to effective multiple therapy and slowing down the
development of HIV resistant strains.
[0169] The limited success in growing reverse transcriptase
crystals by conventional techniques has prompted a search for
alternative methodologies and a certain amount of success has been
accomplished with triple complexing of HIV reverse transcriptase
with Fab fragments and DNA (A. Jacobo Molina et al. Proc. Nat'l.
Acad, Sci. USA 90:6320 (1993)). However, the need for rapid and
robust preparation of transcriptases for structural determination
motivates the search for simpler and more effective cocrystallizing
stabilizers.
[0170] Accordingly, a further aspect of the invention provides a
method for stabilizing HIV-1 reverse transcriptase for
crystallization comprising the steps of contacting a, preferably
pure, solution of the transcriptase with a stabilizing amount of a
compound of Formula 1A and cocrystallizing the resulting complex by
conventional techniques such as the equilibrated hanging drop
method or vapour diffusion method. Alternatively, ready seeded
incipient crystals of transcriptase can be stabilized by
conventional soak techniques to produce crystals having superior
regularity and a longer working life.
[0171] Preferably, the compound of Formula 1A is present in a
slight molar excess, for example a twofold molar excess, over the
concentration of the transcriptase, however the concentration of
the compound of the invention can vary within broad margins, for
example between 1:10 (stabilizer:transcriptase) to 10:1.
[0172] Conventional crystallizing buffers, pH and osmolality
regulators, etc. may be used in the mother liquor as are
exemplified in A. J. McPherson et al., Methods Biochem. Anal.
23:249-345 (1979).
[0173] TEST II Co-crystallization of HIV Reverse Transcriptase.
[0174] A crystallization buffer comprising 14% PEG, 0.1M Tris, pH
8.5, 0.36 mM HIV-1 reverse transcriptase (216 fragment) and 0.4 mM
of the compound
N-(2-imidazo-4-yl)ethyl)-N'-(5-bromopyrid-2-yl)-urea (Example 36)
is subject to hanging drop crystallization (T. Unge et al. Aids
Res. & Human Retroviruses 6:1297 (1990). The resulting crystals
are significantly larger (elongate prisms 2-4 mm long) and
macroscopically more regular than those crystallized without the
compound of the present invention. Preliminary results indicate
that the useful exposure life is in excess of 10 hours, making the
crystals amenable to X-ray crystallography to at least 3.5 .ANG., a
threefold improvement.
[0175] Alternatively, co-crystallization can be effected by vapour
diffusion, wherein a purified HIV-1 reverse transcriptase is
prepared, e.g., in a crystallizing buffer consisting of 25 mM bis
tris propane, pH 7, 50 mM ammonium sulphate, 0.1% (w/v)
.beta.-octyl glucoside, 5% (v/v) polyethylene glycol 8 000 and 0.1%
sodium azide. A twofold molar excess of the compound of the
invention over the concentration of transcriptase is added and
allowed to complex. Crystals are grown by vapour diffusion against
a solution which is double in the concentration of all components
except the transcriptase.
[0176] Preliminary results suggest that the resulting crystals can
have dimensions in excess of 3.0.times.0.6.times.0.4 mm and a
useful exposure lifetime of greater than 10 hours, enabling X-ray
crystallography down to 3.5 .ANG.. This method should be contrasted
against crystals grown without co-crystallization which are
irregular and only amenable to X-ray crystallography to 9.4 .ANG.
resolution (T. Unge et al. Aids Res. & Human Retroviruses
6:1297 (1990)).
[0177] The following examples further illustrate the compounds of
the present invention and methods for the synthesis. The examples
are not intended to be limiting to the scope of the invention in
any respect and should not be so construed. In the descriptions of
the preparations, the reference to example numbers are those found
in Patent Application WO 93/03022 having a publication date of Feb.
13, 1993, hereby incorporated by reference.
Preparation 1
(+,-)-N-(cis-2-(2,6-Difluorophenyl)-cyclopropyl)-N'-(5-ch
loropyrid-2-yl)-thiourea
[0178] The title compound was obtained starting from
2,6-difluorobenzaldehyde (purchased from Aldrich) which was
converted to cis-2,6-difluorophenylcyclopropylamine according to
the procedure in Example 375 and then condensed with the product
from Example 393 according to the procedure in Example 370;
.sup.1H-NMR (250MHz, DNSO-D6). d 1.3-1.4 (q, 1H) 1.6-1.7 (q, 1H)
2.3-2.4 (q, 1H), 3.8-3.9 (m, 1H) 7.2-7.3 (m, 3H) 7.4-7.5 (m, 1H)
7.7 (d, 1H) 7.8-7.9 (m, 1H) 10.7010.8 (s, 1H) 11.0-11.1 (d,
1H).
Preparation 2
(+)-N-[cis-2-(2-chloro-3-ethoxy-6-fluorophenyl)-cyclopropyl]-N'-(5-chloro--
2-pyridyl) thiourea
[0179] The starting material
(.+-.)-cis-2-(2-chloro-3-ethoxy-6-fluoropheny- l)cyclopropylam ine
was prepared from 2-chloro-4-fluorophenol in a manner analogous to
Examples 362, 375 and 348.
[0180] 5-Chloropyrid-2-ylisothiocyanate was prepared as in Example
374 and then condensed with
(.+-.)-cis-2-(2-chloro-3-ethoxy-6-fluorophenyl)cyclop- ropylam ine
in a manner analogous to Example 370 to give the titled
product.
[0181] .sup.1H-NMR (250 MHz, CDCl3) d 11.25 (br s, 1H), 9.09 (br s,
1H), 7.72 (d, 1H), 7.49 (dd, 1H), 6.94 (t, 1H), 6.84 (dd, 1H), 6.74
(d, 1H) 4.12 (q, 2H), 3.67-3.57 9 (m, 1H), 2.23 (q, 1H), 1.78-1.68
(m, 1H), 1.55 (t, 3H), 1.44-1.36 (m, 1H).
[0182] 13C-NMR (62.9 MHz, CDCL3) d 180.6, 156.6 (d), 114.2 (d),
112.8, 118.8 (d), 65.0, 32.7, 16.5, 15.7 (d), 14.7. Anal. calcd.
for C.sub.17H.sub.16Cl.sub.2FN.sub.3OS: C, 51.01; H, 4.03; N,
10.5.
[0183] Found C, 50.9; H, 3.9; N, 10.25.
Preparation 3
N-(2-(2-Chloro-3-ethoxy-6-fluorophenethyl)-N'-(2-(5-bromo)pyridyl)-thioure-
a
[0184] The starting material
2-chloro-3-ethoxy-6-fluorophenethylamine was prepared from
2-chloro-3-ethoxy-6-fluorobenzaldehyde in a manner analogous to
Example 151 and 2-chloro-3-ethoxy-6-fluorobenzaldehyde was prepared
from 2-chloro-4-fluorophenol in a manner analogous to Example 362.
2-Chloro-3-ethoxy-6-fluorophenethylamine was condensed with the
product of Example 392, using the procedure of Example 411 to gove
the titled product.
[0185] .sup.1H-NMR (250 MHz, CDCl.sub.3): d 11.30 (br s, 1H, NH),
9.35 (s, 1H, NH), 8.08 (d, 1H, pyridine), 7.69 (dd, 1H, pyridine),
6.93-6.76 (m, 3H, phenyl, pyridine), 4.11-4.00 (m, 4H CH.sub.2--NH,
OCH.sub.2CH3), 3.24 (t, 2H, phenyl-CH.sub.2), 1.46 (t, 3H,
OCH.sub.2CH3). .sup.13C-NMR (250 MHz, CHC.sub.13): 179, 157, 153,
151, 150, 146, 140, 125, 124, 113, 112, 111, 65, 44, 25, 14.
[0186] Anal. clcd, for C.sub.16H.sub.16BrC1FN.sub.3OS: C, 44.4; H,
3.7; N, 9.7
[0187] Found: C, 44.7; H, 3.9; N, 9.3.
EXAMPLE 1
N-(2-Methoxyphenethyl)-N'-[2-(4-cyano)thiazolyl] urea
[0188] 34
[0189] A solution of N-(imidazolyl)-2-amino-4-cyanothiazole (2.0 g,
9.12 mmol) and 2-methoxyphenethylamine (1.4 g, 9.12 mmol, 1.3 mL)
in N,N-dimethylformamide (20 mL) was stirred at 90-95.degree. C.
After 2 h, the solution was cooled to room temperature, poured into
ethyl acetate and washed with 0.1N hydrochloric acid (2.times.),
water (2.times.), and brine.
[0190] The organic layer was dried over magnesium sulfate, filtered
and concentrated. The solid was recrystallized (50% ethyl
acetate/hexames) providing 1.5 g (54%) of the titled product as
white solid: mp 174.degree. C.; IR (KBr, cm.sup.-1) 3379, 3177,
3110, 3049, 2960, 2230, 1680, 1578, 1494, 1450, 1305, 1245, 1125;
.sup.1H NMR (300 MHZ, DMSO-d.sub.6) 10.82 d(s, 1H), 8.11 (s, 1H),
7.19-7.08 (m, 2H), 6.92 (d, J=7 Hz, 1H), 6.83 (t, J=7 Hz, 1H), 6.52
(s, 1H), 3.73 (s, 3H), 3.32-3.27 (m, 2H), 2.69 (t, J=7.5 Hz, 2H);
MS (FD) m/e 302 (M+); UV (EtOH) 269 nm (.epsilon.=9170), 243 nm
(.epsilon.=9394), 212 nm (.epsilon.=35263).
[0191] Anal. Calc'd. for C.sub.14H.sub.14N.sub.4O.sub.2S: C, 55.62;
H, 4.67; N, 18.53. Found: C, 55.88; H, 4.76; N, 18.28.
EXAMPLE 2
N-(3-Methoxyphenethyl)-N'-[2-(4-ethyl)thiazole]urea
[0192] 35
[0193] A solution of N-(imidazoyl)-2-amino-4-ethylthiazole (2.1 g,
9.4 mmol) and 3-methoxyphenethylamine (9.4 mmol, 1.4 mL) in
N,N-dimethylformamide (20 mL) was stirred at 90-95C. After 3 h, the
solution was allowed to cool to room temperature, was poured into
ethyl acetate and washed with 0.1N hydrochloric acid (2.times.),
water (2.times.), and brine. The organic layer was dried over
magnesium sulfate, filtered, and concentrated. The solid was
recrystallized (50% ethyl acetate/hexanes) providing 1.5 g (52%) of
the titled product as a white solid: mp 88-89.degree. C.; IR (KBr,
cm.sup.-1) 3236, 3011, 2974, 1673, 1563, 1456, 1261, 1155, 1044;
.sup.1H NMR (300 MHZm CDCl.sub.3) d 10.03 (br s, 1H), 7.23-7.17 (m,
1H), 6.83-6.29 (m, 3H), 6.77 (s, 1H), 3.76 (s, 3H), 3.62-3.52 (m,,
2H), 2.85 (t, J=6.9 Hz, 2H), 2.59 (q, J=7.4 Hz, 2H), 1.17 (t, J=7.5
Hz, 3H); MS (FD) m/e 305 (M.sub.+)' UV (EtOH) 266 nm (=11647), 202
nm (=28250).
[0194] Anal. Calc'd. for C.sub.15H.sub.19N.sub.3O.sub.2S:C, 58.99;
H, 6.27; N, 13.76. Found: C, 58.93; H, 6.35; N, 13.79.
EXAMPLE 3
N-[3-Methoxyphenethyl]-N'-[2-N-methyl-4-ethyl-thiazolyl]urea
[0195] 36
[0196] To a solution of 2-N-methyl-4-ethyl thiazole (9.2 g, 64.7
mmol) in 60 mL CH.sub.3CN was added 1,1-carbonlydiimidazole (10.5
g, 64.7 mmol) and 40 mL CH.sub.3CN. The solution was heated to
60-65_C for two days. The CH3CN was removed in vacuo leaving a
brown oil that solidified upon standing. This solid (2.36 g, 10
mmol) was dissolved in DMF (40 mL), treated with
3-methoxyphenethylamine (1.5 g, 10 mmol, 1.46 mL), and stirred at
85.degree. C. overnight. After cooling to room temperature, the
reaction was poured into EtOAc and washed with 0.1N HCl (1.times.),
H.sub.2O (3.times.), and brine (1.times.). The organics were dried
over Na.sub.2SO.sub.4, filtered and concentrated resulting in a
yellow oil which solidified upon standing.
[0197] The solid was purified by flash chromatography on silica gel
(30% EtOAc/hexanes) to provide 1.05 g (33%) of the titled product
as a yellow oil. IR (KBr, cm-1) 3010, 2975, 1673, 1557, 1512, 1339,
1318, 1260, 1105; .sup.1H NMR (300 MHz, CDCl3) d 9.69 (br s, 1H),
7.25-7.18 (m, 1H), 6.85-6.73 (m, 3H), 6.36 (s, 1H), 3.76 (s, 3H),
3.68-3.62 (m, 2H), 3.38(s, 3H), 2.87 (t, J=7 Hz, 2H), 2.54 (q,
J-7.4 Hz, 2H), 1.15 (t, J=7.5 Hz, 3H); MS (FD) m/e 319(M+); UV
(EtOH) 267 nm (.epsilon.=11892), 202 nm (.epsilon.=27796).
[0198] Anal. Calc'd. for C.sub.16H.sub.21N.sub.3O.sub.2S: C, 60.16;
H, 6.65 N, 13.16. Found: C, 60.08; H, 6.53; N, 12.95.
EXAMPLE 4
(+,-)-N-(cis-2(2,6-Difluorophenyl)-cyclopropyl)-N'-(5-chloropyrid-2-yl)-ur-
ea
[0199]
N-(cis-2-(2,6-Difluorophnyl)-cyclopropyl)-N'-(d-chloropyrid-2-yl)-t-
hiourea ((20 mg, 0.06 mmol) was dissolved in dioxane (3 ml) and
water (0.5 ml); N-bromosuccinimide (40 mg, 0.22 mmol) in dioxane (1
ml) was added at ambient temperature. After 10 minutes the solution
was diluted with dichloromethane (10 ml) and washed with dilute
aqueous sodium hydroxide solution (5 ml, 15%). The organic phase
was dried with sodium sulphate, filtered and evaporated. The
residue was purified by flash-chromatography on silica-gel by
elution with a mixture of ethyl acetate and n-hexane (1:1) to yield
the title compound as a white solid. N-NMR (250 MHz, CHCl3-MeOD4).
d 1.2-1.3 (m, 1H) 1.4-1.5 (m, 1H) 2.1-2.2 (q, 1H) 3.2-3.3 (m, 1H)
6.9-7.0 (m, 3H) 7.2-7.3 (m, 1H) 7.5-7.6 (m, 1H) 7.7-7.8 )d,
1H).
EXAMPLE 5
N-(2-Chloro-3-ethoxy-6-fluorophenethyl)-N'-(5-bromopyrid-2-yl)-urea
[0200] The title compound was obtained from
N-(2-Chloro-3-ethoxy-6-fluorop-
henethyl)-N'-(5-bromopyrid-2-yl)-thiourea according to the
procedure in Example 1.
[0201] 1H-NMR (250 MHz, DMSO-D6). d 1.4-1.5 (t, 3H) 3.0-3.1 (t, 2H)
3.5-3.6 (q, 2H) 4.1-4.3 (q, 2H) 7.0-7.3 (m, 2H) 7.5-7.6 (d, 1H)
7.7-7.8 (m, 1H) 7.9-8.0 (d, 1H) 8.4 (s, 1H) 9.2-9.3 (bs, 1H).
EXAMPLE 6
(+,-)-N-(cis-2-(2-chloro-3-ethoxy-6-fluorophenyl)-cyclopropyl)-N'-(5-chlor-
opyrid-2-yl)-urea
[0202] The title compound was obtained from
N-(cis-2-(2-chloro-3-ethoxy-6--
fluorophenyl)-cyclopropyl)-N'-(5-chloropyrid-2-yl)-thiourea
according the the procedure in Example 1.
[0203] 1H-NMR (250 MHz, DMSO-D6). d 1.1-1.2 (m, 1H) 1.4-1.5 (t, 3H)
1.6-1.7 (q, 1H) 2.1-2.2 (q, 1H) 3.3-3.4 (m, 1H) 4.2-4.4 (q, 2H)
7.2-7.3 (m, 2H) 7.4-7.5 (d, 1H) 7.7-7.8 (m, 1H) 7.9 (d, 1H) 8.0-8.1
(bs, 1H) 9.4 (bs, 1H).
EXAMPLE 7
N-(2-(2,6-difluoro)-phenethyl)-N'-(2-(5-chloro)-pyridinyl) urea
[0204] To a solution of 2.0 g (11 mmol) 2,6-Difluorocinnamic acid
in ethanol (50 ml) was added about 100 mg of 5% of palladium on
carbon. The reaction mixture was hydrogenated at atmospheric
pressure until 250 ml of hydrogen was absorbed. The mixture was
filtered and the solvent evaporated. Careful drying gave 1.87 g of
a crude product which was used unpruified in the next step.
[0205] To solution of 1.87 g (10 mmol) of the above described crude
product in dry toluene (20 ml) was added 1.53 ml (11 mmol) of
triethylamine and 2.37 ml (11 mmol) of diphenylphosphoryl azide.
The reaction mixture was stirred for 5 hours at room temperature,
and then refluxed for 30 minutes. Thereafter, 1.54 g (12 mmol) of
2-amino-5-chloropyridine was added and the reaction mixture was
stirred over night at +100.degree. C. After cooling, EtOAc and
brine were added. The organic phase was separated and dried over
sodium sulfate. Evaporation of the solvent gave 3.75 g of crude
product. 1.0 g of this material was purified on a silica gel column
using ether 1 and hexanes 1 as the eluent. This procedure gave only
a partial purification of the title compound and a final
purification was performed on an alumina column which was eluted
with THF 1 hexanes 2 followed by MeOH. 129 mg of the title compound
was obtained.
[0206] Analysis: Calculated: C 53.95, H 3.9, N 13.5. Found: C 53.8,
H 3.9, N 13.6
[0207] MP: 173.0-175.0.degree. C.
[0208] 1H-NMR (CDCl.sub.3) d 3.00 (t, 2H), 3.62 (q, 2H), 6.80-6.90
(m, 3H), 7.10-7.23 (m, 1H), 7.53 (dd, 1H), 8.01 (d, 1H), 9.17
(broad t, 1H), 9.39 (broad s, 1H).
[0209] 13C-NMR (DMSO-d.sub.6) d 22.91, 38.63, 111.35 (d, 2C),
112.91, 114.56 (t, 1C), 122.79, 128.85 (t, 1C), 138.03, 145.10,
152.11, 154.50, 161.34 (dd, 2C).
EXAMPLES 8 & 9
N-(2-Phenethyl)-N'-(5-bromopyrid-2-yl)-urea and
N-(2-Phenethyl)-N'-(3,5-di- bromopyrid-2-yl)-urea
[0210] A solution of 10.0 g (37.3 mmole) of
N-(imidazoyl)-2-amino-5-bromop- yridine and 4.3 g (35.5 mmole) of
phenethylamine in N, N-dimethylformamide (100 ml) was stirred at
90-95.degree. C. for 2.5 hrs. The solution was cooled to room
temperature, poured into ethyl acetate, and washed with water
(5.times.), saturated sodium bicarbonate (3.times.), and brine
(1.times.). The organic layer was dried over sodium sulfate,
filtered, and concentrated. The solid was chromatographed over a
flash silica gel column (2:1 hexanes/ethyl acetate) to provide
isolation of two components. The first component was isolated as a
white solid, and was found to be the dibromopyridyl urea, 35 mg: mp
145-147.degree. C.; IR (KBr, cm.sup.-1) 3400, 3300, 3000, 1683,
1474, 1364, 1285, 1230, 1035, 735, 700; .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta.8.45 (br.m., 1H), 8.40 (d, 1H), 8.25 (d, 1H),
8.00 (br.s, 1H), 7.20-7.40 (m, 5H), 3.45 (q, 2H), 2.80 (t, 2H); MS
(FD) m/e 399 (Mt). Anal. Calc'd. for
C.sub.14H.sub.13Br.sub.2N.sub.3O H.sub.2O; C, 41.65; H, 3.34; N,
10.41. Found: C, 41.62; H, 3.26; N, 10.32.
[0211] The second component was isolated as a white solid, filtered
with the aid of ethyl ether, and was found to be the
monobromopyridyl urea, 4.2 g (37.5%): mp 164-167.degree. C.; IR
(KBr, cm.sup.-1) 3275, 3200, 3000, 1686, 1540, 1363, 1302, 1230,
830, 701, 513; .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.9.30
(br.s., 1H), 8.20 (d, 1H), 7.90 (dd, 1H), 7.70 (br.t., 1H),
7.20-7.38 (m, 5H), 3.40 (q, 2H), 2.80 (t, 2H); MS (FD) m/e 319
(Mt). Anal. Calc'd. for C.sub.14H.sub.14BrN.sub.3O: C, 52.52; H,
4.41; N, 13.12. Found: C, 52.78; H, 4.56; N, 13.18.
EXAMPLE 10
N-(3-Fluoropyrid-2-yl)eth-2yl-N'-(5-bromopyrid-2-yl)-urea
[0212] A solution of 400 mg (1.13 mmole) of
N-(3-fluoropyrid-2-yl)eth-2-yl- -N'-(5-bromopyrid-2-yl)-thiourea
(see Example 920 of WO93/03022) in 11.5 ml of glacial acetic acid
was treated with 0.8 ml (7.83 mmole) of 30% hydrogen peroxide.
After stirring 1.5 hrs at room temperature, the solution was
concentrated and the residue was partitioned between ethyl acetate
and saturated aqueous sodium bicarbonate (1.times.). The organic
layer was back washed with brine (1.times.) and dried over sodium
sulfate. Purification by flash silica gel chromatography (50% ethyl
acetate/hexanes) gave 75 mg (19.6%) of the titled compound as a
white solid: mp 151-154.degree. C.; IR (KBr, cm.sup.-1) 3200, 3100,
3000, 1685, 1557, 1467, 1360, 1290, 1230, 839; .sup.1H NMR (300
MHz, DMSO-d.sub.6) .delta.9.30 (br. s., 1H), 8.40 (d, 1H), 8.22 (d,
1h), 7.85 (dd, 1H), 7.79 (br.t., 1H), 7.70 (dt, 1H), 7.45 (d, 1H),
7.30-7.40 (m, 1H), 3.58 (q, 2H), 3.00 (t, 2); MS (FD) m/e 340 (Mt).
Anal. Calc'd. for C.sub.13H.sub.12BrFN.sub.4O: C, 46.04; H, 3.57;
N, 16.52. Found: C, 46.25; H, 3.66; N, 16.54.
EXAMPLE 11
(+,-)-n-(cis-2-Phenylcyclopropyl)-N'-(5-chloropyrid-2-yl)-urea
[0213] The titled compound was obtained from
(+,-)-N-(cis-2-phenylcyclopro-
pyl)-N'-(5-chloropyrid-2-yl)-thiourea (see Example 372 of WO
93/03022) according to the procedure in Example 10: mp
178-180.degree. C.; IR (KBr, cm.sup.-1) 3200, 3000, 1709, 1544,
1287, 698; .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.9.20 (br.s.,
1h), 7.93 (d, 1H), 7.80 (dd, 1H), 7.60 (br.s., 1H), 7.18-7.38 (m,
6H), 3.00-3.10 (m, 1H), 2.20-2.30 (q, 1H9, 1.20-1.30 (q, 1H),
1.00-1.10 (q, 1H); MS (FD) m/e 287 (Mt). Anal. Calc'd. for
C.sub.15H.sub.14ClN.sub.3O: c, 62.61; H, 4.90; N, 14.60. Found: C,
62.35; H, 4.98; N, 14.56.
EXAMPLE 12
N-(2-Cyclohexen-1-yl) ethyl-N'-(5-bromopyrid-2-yl)-urea
[0214] The titled compound was obtained from
N-(2-cyclohexen-1-yl)ethyl-N'- -(5-bromopyrid-2-yl)-thiourea (see
Example 189 of WO 93/03022) according to the procedure in Example
10: mp 128-130.degree. C.; IR (KBr, cm.sup.-1) 3200, 2930, 1682,
1550, 1364, 1292, 1234, 836; .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta.9.35 (s, 1H), 8.25 (d, 1H), 7.87 (dd, 1H), 7.60 (br.t., 1H9,
7.45 (d, 1H), 5.43 (br.s., 1H), 3.20 (q, 2H), 2.10 (t, 2H),
1.85-2.00 (m, 4H), 1.45-1.60 (m, 4H); MS (FD) m/e 325 (Mt). Anal.
Calc'd. for C.sub.14H.sub.18BrN.sub.3O: C, 51.86; H, 5.60; N,
12.96. Found: C, 52.07; H, 5.34; N, 12.68.
EXAMPLE 13
N-(2-(2,6-Difluoro-3-methoxyphenethyl)-N'-(5-bromopyrid-.sup.2-yl)-urea
[0215] The titled compound was obtained from
N-(2-(2,6-difluoro-3-methoxyp-
henethyl)-N'-(5-bromopyrid-2-yl)-thiourea (see Example 411 of WO
93/03022) according to the procedure in Example 10: mp
198-200.degree. C.; IR (KBr, cm.sup.-1) 3200, 3000, 1686, 1492,
1364, 1223, 1077, 805; .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta.9.30 (s, 1H), 8.20 (d, 1H), 7.85 (dd, 1H), 7.75 (br. t.,
1H), 7.43 (d, 1H), 6.95-7.10 (m, 2H), 3.80 (s, 3H), 3.30 (q, 2H),
2.83 (t, 2H); MS (FD) m/e 385 (Mt). Anal. Calc'd. for
C.sub.15H.sub.14BrF.sub.2N.sub.3O.sub.2: C, 46.65; H, 3.65; N,
10.88. Found: C, 46.53; H, 3.85; N, 10.64.
EXAMPLE 14
N-(2-(2-Chloro-3-ethoxy-6-fluorophenethyl)-N'-(pyrazo-3-yl)-urea
[0216] The starting material
2-chloro-3-ethoxy-6-fluorophenethylamine was prepared from
2-chloro-3-ethoxy-6-fluorobenzaldehyde as in Example 151 of WO
93/03022 and 2-chloro-3-ethoxy-6-fluorobenzaldehyde was prepared
from 2-chloro-4-fluorophenol as in Example 362 of WO 93/03022. The
titled compound was prepared from
2-chloro-3-ethoxy-6-fluoro-phenethylamine and
N-(imidazoyl)-3-aminopyrazole according to Examples 8 & 9: mp
110-115.degree. C.; IR (KBr, cm.sup.-1) 3200, 3000, 1683, 1469,
1370, 1232, 1087, 829; .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta.12.10 (br.s., 1H), 8.70 (s, 1H), 7.50 (br.s., 1H), 7.10-7.20
(m, 1H), 6.90-7.05 (m, 2H), 6.05 (br.s., 1H), 4.07 (q, 2H), 3.30
(q, 2H), 2.93 (br.t., 2H), 1.35 (t, 2H); MS (FD) m/e 326 (Mt).
Anal. Calc'd.. for C.sub.14H.sub.16ClFB.su- b.4O.sub.2: C, 51.46.;
H, 4.94; N, 17.15. Found: C, 51.71; H, 4.81; N, 17.29.
EXAMPLE 15
N-(2-(2,6-Difluorophenethyl)-N'-(5-bromopyrid-2-yl)-urea.
[0217] The titled compound was prepared from
N-(2-2,6-difluorophenethyl)-N- '-(5-bromopyrid-2-yl)-thiourea (see
Example 340 of WO 93/03022) as in Example 10: mp 185-187.degree.
C.; IR (KBr, cm.sup.-1) 3094, 1680, 1468, 1237, 1000, 830, 775;
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.9.30 (s, 1H), 8.20 (d,
1H), 7.86 (dd, 1H), 7.73 (br.t., 1H), 7.45 (d, 1H), 7.30-7.40 (m,
1H), 7.00-7.10 (t, 2H), 3.40 (q, 2H), 2.82 (t, 2H). MS (FD) m/e 355
(Mt). Anal. Calc'd. for C.sub.14H.sub.12BrF.sub.2N.sub.3O: C,
47.21; H, 3.40; N, 11.80. Found: C, 46.93; H, 3.61; N, 11.82.
EXAMPLE 16
N-(2-Phenethyl)-N'-(pyrimid-4-yl)-urea
[0218] The titled compound was prepared from
N-(imidazoyl)-4-aminopyrimidi- ne and phenethylamine according to
the procedure in Examples 8 & 9: mp 162-164.degree. C.; IR
(KBr, cm.sup.-1) 3200, 3113, 3045, 2944, 1680, 1589, 1306, 993,
761; .sup.1H NMR (300 MHZ, DMSO-d.sub.6) .delta.9.60 (s, 1H), 8.65
(s, 1H), 8.45 (d, 1H), 7.75 (t, 1H), 7.50 (d, 1H), 7.20-7.40 (m,
5H), 3.40 (q, 2H), 2.78 (t, 1H); MS (FD) m/e 242 (Mt). Anal.
Calc'd. for C.sub.13H.sub.14N.sub.4O: C, 64.45; H, 5.82; N, 23.13.
Found: C, 64.67; H, 6.06; N, 23.24.
EXAMPLE 17
N-(2-Phenethyl)-N'-(5-bromopyrimid-2-yl) -urea
[0219] The titled compound was prepared from
N-(2-Phenethyl)-N'-(5-bromopy- rimid-2-yl)-thiourea (see Example
222 of WO 93/03022) according to the procedure in Example 10: mp
205-208.degree. C.; IR (KBr, cm.sup.-1) 3309, 3045, 2972, 1692,
1564, 1504, 1415, 1285, 832; .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta.10.00 (s, 1H), 8.70 (t, 1H), 8.65 (s, 2H), 7.20-7.40 (m,
5H), 3.45 (q, 2H), 2.80 (t, 2H); MS (FD) m/e 320 (Mt). Anal.
Calc'd. for C.sub.13H.sub.13BrN.sub.4O 0.5 H.sub.2O: C, 47.30; H,
4.24; N, 16.97. Found: C, 47.10; H, 3.94; N, 16.94.
EXAMPLE 18
N-(2-Phenethyl)-N'-(6-chloropyridaz-3-yl)-urea
[0220] The titled compound was prepared from
N-(2-Phenethyl)-N'-(6-chlorop- yridaz-3-yl)-thiourea (see Example
336 of WO 93/03022) as in Example 10: mp 222-225.degree. C.; IR
(KBr, cm.sup.-1) 3341, 2980, 1709, 1554, 1420, 1251, 1140, 696;
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.9.80 (s, 1H), 8.05 (d,
1H), 7.75 (d, 1H), 7.10-7.40 (m, 6H), 3.40 (q, 2H), 2.80 (t, 2H);
MS (FD) m/e 276 (Mt). Anal. Calc'd. for C.sub.13H.sub.13ClN.sub.4O
0.25 H.sub.2O: C, 55.55; H, 4.80; N, 19.93. Found: C, 55.36; H,
4.67; N, 19.95.
EXAMPLE 19
N-(2-Phenethyl)-N'-(4-bromopyrazo-3-yl)-urea
[0221] The titled compound was prepared from
N-(imidazoyl)-2-phenethylamin- e and 3-amino-4-bromo-pyrazole as in
Examples 8 & 9: mp 132-135.degree. C.; IR(KBr, cm.sup.-1) 3351,
3271, 1712, 1530, 1386, 1238, 1078, 700; .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta.8.17 (s, 1H), 7.80 (t, 1H), 7.20-7.38 (m, 5H),
5.40 (s, 2H), 3.45 (q, 2H), 2.83 (t, 2H); MS (FD) m/e 308 (Mt).
Anal. Calc'd. for C.sub.12H.sub.13BrN.sub.4O: C, 46.62; H, 4.24; N,
18.12. Found: C, 46.65; H, 4.14; N, 17.93.
EXAMPLE 20
N[(4-Ethoxy-3-fluoropyrid-2-yl)eth-2-yl]-N'-(5-bromopyrid-2-yl)-urea
20a)
N-[(3-Fluoropyrid-2-yl)eth-2-yl]-2,5-dimethylpyrrole
[0222] A mixture of 3.6 g (25.70 mmole) of
2-(2-aminoethyl)-3-fluoropyridi- ne, 3.0 g(26.3 mmole) of
2,5-hexanedione, 0.4 ml of glacial acetic acid, and 100 ml of
toluene was refluxed with a Dean-Stark trap for 3 hrs. The mixture
was cooled to room temperature and extracted with saturated aqueous
sodium bicarbonate (1.times.) and brine (1.times.). The organic
layer was dried over sodium sulfate, filtered, and concentrated to
a solid. Recrystallization from ethyl ether/hexanes provided the
titled compound as an off-white solid; 5.0 g (90%): mp
82-84.degree. C.; IR (KBr, cm.sup.-1) 3075, 2931, 1448, 1407, 1112,
755; .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.8.40 (d, 1H), 7.66
(dt, 1H), 7.38 (m, 1H), 5.60 (s, 2H), 4.05 (t, 2H), 3.05 (dt, 2H),
2.10 (s, 6H); MS (FD) m/e 218 (Mt). Anal. Calc'd. for
C.sub.13H.sub.15FN.sub.2: C, 71.53; H, 6.93; N, 12.83. Found: C,
71.73; H, 7.05; N, 13.04.
20b)
N-[(3-Fluoro-4-iodopyrid-2-yl)eth-2-yl]-2,5-dimethyl-pyrrole
[0223] A dry 250 ml 3-neck roundbottom flask equipped with a
thermometer, rubber septum, magnetic stirrer, and nitrogen inlet
was charged with 30 ml of anhydrous tetrahydrofuran and 2.1 ml
(14.67 mmole) of dry diisopropylamine. The stirring solution was
cooled to -78.degree. C. afterwhich 9.5 ml (15.11 mmole) of 1,6
n-butyllithium in hexanes was added over 10 minutes by syringe. The
solution was stirred 30 minutes longer and 3.2 g (14.67 mmole) of
N-[(3-Fluoropyrid-2-yl)eth-2-yl]-2,5-di- methylpyrrole in 30 ml of
dry tetrahydrofuran was added dropwise via syringe over 30 minutes,
keeping the internal temperature below -65.degree. C. After
stirring 30 minutes longer, a solution of 4.0 g (15.80 mmole) of
iodine in 30 ml of dry tetrahydrofuran was added by canula over 10
minutes. The mixture was warmed to room temperature (a color
change, quenching, occurred at -30.degree. C.) and stirred
overnight under nitrogen. The solution was diluted to 200 ml with
ethyl ether, extracted with aqueous saturated sodium bicarbonate
(IX), brine (IX), and dried over sodium sulfate. Filtration and
concentration gave a dark solid which was purified by flash silica
gel chromatography (20% ethyl acetate in hexanes) to give the
titled compound as a white solid, 3,9 g (77%): mp 156-158.degree.
C.; IR (KBr, cm.sup.-1) 3000, 2950, 1574, 1408, 752; .sup.1H NMR
(300 MHz, DMSO-d.sub.6) .delta.8.05 (d, 1H), 7.85 (t, 1H), 5.60 (s,
2H), 4.05 (t, 2H), 3.00 (dt, 2H), 2.10 (s, 6H); MS (FD) m/e 344
(Mt). Anal. Calc'd. for C.sub.13H.sub.14FIN.sub.2: C, 45.37; H,
4.10; N, 8.14. Found: C, 45.07; H, 4.03; N, 8.06.
20c)
N[(4-Ethoxy-3-fluoropyrid-2-yl)eth-2-yl]-2,5-dimethyl-pyrrole
[0224] A solution of sodium ethoxide was initially prepared by
dissolving 2.0 g (87 mmole) of sodium metal in 300 ml of anhydrous
absolute ethanol. To the stirring solution thus prepared was added
3.5 g (10.2 mmole) of
N-[(3-Fluoro-4-iodopyrid-2-yl)eth-2-yl]-2,5-dimethyl-pyrrole and
the mixture was refluxed under nitrogen for 4 days. The solution
was cooled, diluted to 1 liter with water, and extracted with ethyl
acetate (2.times.). The organic layer was dried over sodium
sulfate, filtered, and concentrated to a resin. Chromatography over
flash silica gel (30% ethyl acetate in hexanes) gave initially 320
mg (8.5%) of
N-[(4-Iodo-3-ethoxypyrid-2-yl)-eth-2-yl]-2,5-dimethylpyrrole:
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.7.95 (d, 1H), 7.62 (d,
1H), 5.80 (s, 2H), 4.10 (t, 2H), 4.00 (q, 2H), 3.15 (t, 2H), 2.20
(s, 6H), 1.45 (t, 3H); MS (FD) m/e 370 (Mt). Further elutlon
provided the titled compound, 1.7 g (64%): IR (KBr, cm.sup.-1)
2991, 1608, 1495, 1309, 1075; .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta.8.22 (d, 1H), 7.15 (t, 1H), 5.60 (s, 2H), 4.20 (q, 2H), 4.10
(t, 2H), 2.98 (dt, 2H), 2.10 (s, 6H), 1.38 (t, 3H): MS (FD) m/e 262
(Mt). Anal. Calc'd. for C.sub.15H.sub.19FN.sub.2O 0.25 H.sub.2O: C,
67.55; H, 7.31; N, 10.50. Found: C, 67.56; H, 7.35; N, 10.46.
20d)
2-(2-Aminoethyl)-4-ethoxy-3-fluoropyridine
[0225] A mixture of 825 mg (3.15 mmole) of
N-[(4-Ethoxy-3-fluoropyrid-2-yl- )eth-2-yl]-2,5-dimethylpyrrole,
1.4 g (20.1 mmole) of hydroxylamine hydrochloride, 700 mg (12.5
mmole) of potassium hydroxide, 20 ml of ethanol, and 8 ml of water
was refluxed for 3 days under nitrogen. The solution was cooled to
room temperature, concentrated, and partitioned between 50 ml of 2N
hydrochloric acid and ethyl acetate (3.times.). The aqueous layer
was basified with 5N sodium hydroxide, extracted with ethyl acetate
(3.times.), dried over sodium sulfate, and concentrated. The
resulting red oil was found to be the titled compound with a small
amount of 2,5-hexanedione dioxime also present. The oil was used as
such for reaction without further purification: IR (KBr, cm.sup.-1)
2989, 1609, 1495, 1306, 1075; .sup.1H NMR (300 MHz, DMSO-d.sub.6)
.delta.8.18 (d, 1H), 7.05 (t, 1H), 4.20 (q, 2H), 2.60-3.00 (m, 6H),
1.38 (t, 3H); MS (FD) m/e 185 (Mt).
20e)
N-[(4-Ethoxy-3-fluoropyrid-2-yl)eth-2-yl]-N'-(5-bromopyrid-2-yl)-urea
[0226] The titled compound was prepared from
N-(imidazoyl)-2-amino-5-bromo- pyridine and
2-(2-aminoethyl)-4-ethoxy-3-fluoropyridine as in Examples 8 &
9: mp 143-145.degree. C.; IR (KBr, cm.sup.-1) 3221, 2989, 1684,
1475, 1306; .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.9.30 (s,
1H), 8.10 (m, 2H), 7.90 (dd, 1H), 7.78 (br.t., 1H), 7.42 (d, 1H),
7.15 (t, 1H), 4.20 (q, 2H), 3.55 (q, 2H), 2.92 (dt, 2H), 1.35 (t,
3H): MS (FD) m/e 382 (Mt). Anal. Calc'd. for
C.sub.15H.sub.16BrFN.sub.4O.sub.2: C, 47.01; H, 4.21; N, 14.62.
Found: C, 46.80; H, 4.21; N, 14.41.
EXAMPLE 21
(+,-)-N-(cis-2-(3-Nitrophenyl)-cyclopropyl)-N'-(5-chloropyrid-2-yl)-urea
[0227] 3-Nitrostyrene (Aldrich) was converted to
cis-2-(3-nitrophenyl)cycl- opropanecarboxylic acid by the method
described in Example 348 (Patent Application WO 93/03022), with the
exception that the separation of cis-isomer was performed at the
ethyl ester stage using a silica gel column eluted with hexanes 9
and ethyl acetate 1. The acid was then converted to an isocyanate
as described in J. Org. Chem., p. 3511 (1961). The title compound
was obtained by reacting the above described isocyanate with
2-amino-5-chloropyridine as described in Example 7. Analysis:
Calculated: C 54.1, H 3.9, N 16.8. Found: C 53.7, H 3.9, N 16.5.
Mp: 201.0-202.0.degree. C. .sup.1H-NMR (250 MHz, CDCl.sub.3)
.delta.1.20-1.29 (m, 1H), 1.49-1.61 (m, 1H), 2.44 (q, 1H),
3.25-3.33 (m, 1H), 6.69 (d, 1H), 7.38-7.51 (m, 2H), 7.61 (d, 1H),
7.77 (d, 1H), 8.03-8.08 (m, 2H), 8.47 (broad s, 1H), 8.90 (broad s,
1H).
EXAMPLE 22
(+,-)-N-(cis-2-(3-Ethynylphenyl)-cyclopropyl)-N'-(5-chloropyrid-2-yl)-urea
[0228] 3-Bromostyrene (Aldrich) was converted to the ethyl ester of
cis-2-(3-bromophenyl)cyclopropanecarboxylic acid in a process
similar to the one described in Example 21.
[0229] The ethyl ester was then reacted with TMS-acetylene in the
presence of Pd(II) as described in Tetrahedron Letters p. 6403-6406
(1993). After deprotection (LiOH), the resulting
cis-2-(3-ethynylphenyl)-cyclopropaneca- rboxylic acid was
transformed to the title compound according to the procedure in
Example 7. Analysis: Calculated: C 65.5, H 4.5, N 13.6. Found: C
65.6, H 4.4, N 12.9. Mp: 148.5-149.5.degree. C. .sup.1H-NMR (250
MHz, CDCl.sub.3) .delta.1.07-1.14 (m, 1H), 1.38-1.44 (m, 1H),
2.27-2.38 (m, 1H), 3.04 (s, 1H), 3.23-3.31 (m, 1H), 6.77 (d, 1H),
7.20-7.50 (m, 5H), 7.75 (d, 1H), 8.90 (broad s, 1H), 9.20 (broad s,
1H).
EXAMPLE 23
(+,-)-N-(cis-2-(3-Acetylphenyl)-cyclopropyl)-N'-(5-chloropyrid-2-yl)-urea
[0230] A solution of
(+,-)-N-(cis-2-(3-Ethynylphenyl)-cyclopropyl)-N'-(5-c-
hloropyrid-2-yl)-urea of Example 22(130 mg, 0.42 mmole),
H.sub.2SO.sub.4 (10 drops), H.sub.2O (10 drops) and Hg(OAc).sub.2
(100 mg) in acetic acid (4 mL) was stirred at room. temperature for
30 min. The reaction mixture was then poured into an ethyl acetate
and saturated K.sub.2CO.sub.3 solution. The organic layer was dried
(MgSO.sub.4) and the solvent was evaporated to give a residue which
was purified on silica gel using 1:1 acetone: and hexanes as the
eluent providing 43 mg of the title compound.
[0231] Analysis: Calculated: C 61.9, H 4.9, N 12.75. Found: C 61.5,
H 4.7, N 12.1. Mp: 144.0-145.0.degree. C. .sup.1H-NMR (250 MHz,
CDCl.sub.3) .delta.1.15-1.24 (m, 1H), 1.40-1.51 (m, 1H), 2.34-2.47
(m, 1H), 2.55 (s, 3H), 3.23-3.34 (m, 1H), 6.85 (d, 1H), 7.35-7.52
(m, 3H), 7.62 (d, 1H), 7.77-7.90 (m, 2H), 8.90 (broad s, 1H), 9.45
(broad s, 1H).
EXAMPLE 24
(+,-)-N-(cis-2-(3-Aminophenyl)-cyclopropyl)-N'-(5-chloropyrid-2-yl)-urea
[0232] Cis-2-(3-nitrophenyl)cyclopropanecarboxylic acid (355 mg,
1.7 mmole), described in Example 21, was dissolved in 50 ml ethanol
and 20 mg of PtO.sub.2 was added. Hydrogenation at atmospheric
pressure for 1 hr. followed by filtration and evaporation of the
solvent afforded 288 mg of
cis-2-(3-aminophenyl)cyclopropanecarboxylic acid. 177 mg (1 mmole)
of cis-2-(3-aminophenyl)cyclopropanecarboxylic acid was dissolved
in water (1 mL), t-BuOH (1 mL) and 2M NaOH (0.5 mL, 1 mmole). 250
.mu.L ( 1.1 mmole) of di-tert-butyl dicarbonate was then added and
the reaction mixture was stirred at room temperature overnight. The
reaction mixture was poured into ice and water and acidified with
dilute H.sub.2SO.sub.4 and extracted with dichloromethane affording
264 mg of pure BOC protected
cis-2-(3-aminophenyl)-cyclopropanecarboxylic acid. This acid was
transformed to BOC protected title compound according to the
procedure in Example 7. Finally, deprotection with THF 1 and 6M HCl
1 gave the title product. Analysis: Calculated: C 59.5, H 5.0, N
18.5. Found: C 59.5, H 4.9, N 17.2. Mp: 177.5-178.2.degree. C.
.sup.1H-NMR (250 MHz, CDCl.sub.3) .delta.1.00-1.07 (m, 1H),
1.23-1.37 (m, 1H), 2.24-2.33 (m, 1H), 3.15-3.25 (m, 1H), 3.65
(broad s, 2H), 6.59-6.70 (m, 4H), 6.77 (broad s, 1H), 7.11 (t, 1H),
7.45 (dd, 1H), 7.72 (s, 1H), 8.81 (broad s, 1H).
EXAMPLE 25
N-(3-Methoxyphenethyl) -N'-(5-chloropyrid-2-yl) -urea
[0233] Following the condensation procedure described in Example 2
and using N-(imidazoyl)-2-amino-5-chloropyridine instead of
N-(imidazoyl)-2-amino-4-ethylthiazole, resulted in the titled
product. .sup.1H-NMR (250 MHz, CDCl.sub.3) .delta.2.89 (t, 2H),
3.65 (q, 2H), 3.79 (s, 3H), 6.78-6.87 (m, 4H), 7.25-7.28 (m, 1H),
7.52 (dd, 1H), 7.99 (d, 1H), 9.05 (broad s, 1H), 9.12 (broad t,
1H).
EXAMPLE 26
(+,-)-N-(cis-2-(2,6-Difluorophenyl)-cyclopropyl)-N'-(5-cyanopyrid-2-yl)-ur-
ea
[0234] 2,6-Difluorobenzaldehyde (Aldrich) was converted to
cis-2-(2,6-difluorophenyl)cyclopropylamine according to Example 375
(Patent Application WO 93/03022) and then converted to an
isothiocyanate according to Example 374 (Patent Application WO
93/03022). This isothiocyanate was reacted with the anion of
2-amino-5-cyanopyridine to give (+,-)-N-(cis-2-(2,6-Difluorophenyl)
cyclopropyl)-N'-(5-cyanopyrid-2-- yl)-thiourea. 6 mg of this
compound was reacted with NBS as described in Example 4 and a final
purification on alumina, resulted in 0.7 mg of the title product.
Rf: 0.30 Silica gel eluent: diethyl ether.
EXAMPLE 27
(1R,2S)-N-(cis-2-(2,6-Difluorophenyl)-cyclopropyl)-N'-(5-bromopyrid-2-yl)--
urea
[0235]
(1R,2S)-N-(cis-2-(2,6-Difluorophenyl)-cyclopropyl)-N'-(5-bromopyrid-
-2-yl)-thiourea was prepared according to Examples 422, 424 and 427
(Patent Application WO 93/03022) starting with
(+,-)-cis-2-(2,6-difluorop- henyl)-cyclopropylamine instead of
(+,-)-cis-cyclopropylamine. [.alpha.]D.sup.22 of
(1R,2S)-N-(cis-2-(2,6-Difluorophenyl)-cyclopropyl)-N-
'-(5-bromopyrid-2-yl)-thiourea was -47.8.degree. (c=0.54, EtOAc).
Application of the method described in Example 4 using (1R,
2S)-N-(cis-2-(2,6-Difluorophenyl)-cyclopropyl)-N'-(5-bromopyrid-2-yl)-thi-
ourea afforded the title compound. .sup.1H-NMR (250 MHz,
acetone-d.sub.6) .delta.1.20-1.30 (m, 1H), 1.40-1.50 (m, 1H),
2.00-2.20 (m, partly hidden by the signals from the solvent, 1H),
3.25-3.35 (m, 1H), 6.95-7.10 (m, 2H), 7.22 (d, 1H), 7.30-7.50 (m,
1H), 7.79 (dd, 1H), 7.95 (d, 1H), 8.28 (broad s, 1H), 8.60 (broad
s, 1H).
EXAMPLE 28
(1R,2S)-N-(cis-2-(2-chloro-3-ethoxy-6-fluorophenyl)-cyclopropyl)-N'-(5-chl-
oropyrid-2-yl)-urea
[0236] (1R,2S)-cis-2-(2-chloro-3-ethoxy-6-fluorophenyl)
cyclopropylcarboxylic acid was prepared from
2-chloro-3-ethoxy-6-fluorost- yrene, using an asymmetric
cyclopropanation reaction catalyzed by Cu(I)triflate and a chiral
ligand as described by Evans et al. in J.Am.Chem.Soc. 1991, 113,
726-728. This acid was transformed to the title compound according
to the procedure described in Example 7. Analysis: Calculated: C
53.1, H 4.2, N 10.9. Found: C 53.3, H 4.3, N 11.0.
[0237] Mp: 168.5-169.0.degree. C. [.alpha.]D.sup.22-80.00 (c=0.50,
CHCl.sub.3). .sup.1H-NMR (250 MHz, CDCl.sub.3) .delta.1.27-1.35 (m,
1H), 1.53 (t, 3H), 1.55-1.67 (m, 1H), 2.01-2.14 (m, 1H), 3.25-3.29
(m, 1H), 4.09 (q, 2H), 6.72-6.94 (m, 3H), 7.45 (dd, 1H), 7.80 (d,
1H), 8.86 (broad s, 1H), 9.25 (broad s, 1H).
EXAMPLE 29
N-(2-phenethyl)-N'-(thiozol-2-yl)-urea
[0238] 3.38 g phenyl propanoic acid chloride (mw 169, 20 mmole) was
dissolved in 20 ml toluene. A saturated aqueous solution containing
LiN.sub.3 1.25 g (mw 48, 25 mmole) was added under vigorous
stirring at O.degree. C. The mixture was heated to 20.degree. C.
and stirred for 1 hr. The aqueous phase was separated by means of a
silanized filter paper. The organic phase was dried with
Na.sub.2SO.sub.4 and the solvent removed. The residue was
redissolved in 20 ml toluene, and heated to 90.degree. C. for 15
min. The solvent was removed and the raw isocyanate dissolved in 10
ml DMF together with 2 g 2-aminothiazole (mw 100, 20 mmole). This
mixture was stirred at 110.degree. C. for 17 hrs. The solvent was
removed, and the raw product dissolved in 200 ml ethyl acetate,
washed twice with 50 ml 1M hydrochloric acid, and once with 50 ml
water. Crystallisation from ethanol gave fine needles, 2.21 g Mp
162.degree. C. .sup.1H-NMR (250 MHz, DMSO-d.sub.6): 2.86 (t, 2H),
3.00 (t, 2H), 7.25-7.43 (m, 6H), 7.57 (d, 1H).
EXAMPLE 30
N-(2-ethoxy-5,6-difluorophenethyl)-N'-(5-bromopyrid-2-yl)-urea
[0239] 32.5 g 3,4-difluorophenol (Aldrich) was dissolved in 600 ml
acetone. 50 g K.sub.2CO.sub.3 and 45 g iodoethane was added, and
the mixture stirred at 50.degree. C. overnight. The product
1-ethoxy-3,4-difluorobenzene was purified by distillation, and was
then transformed to 2-ethoxy-5,6-difluorobenzaldehyde according to
the procedure in Example 362 of WO 93/03022.
[0240] The methods described in Example 151 of WO 93/03022 was then
used to transform this compound to
2-ethoxy-5,6-difluoro-phenethylamine. The method described in
Example 411 of WO 93/03022 was then used to prepare
N-(2-ethoxy-5,6-difluorophenethyl)-N'-(5-bromopyrid-2-yl)-thiourea.
This product was then oxidised with NBS as Example 4 to provide
N-(2-ethoxy-5,6-difluorophenethyl)-N'-(5-bromopyrid-2-yl)-urea.
.sup.1H-NMR (250 MHz, CDCl.sub.3): 1.4 (t, 3H), 3.0 (q, 2H), 3.95
(q, 2H), 6.5 (m, 1H), 6.8 (d, 1H), 6.95 (dd, 1H), 7.68 (dd, 1H),
8.90 (d, 1H) 8.9 (s, 1H), 9.0 (s, 1H).
EXAMPLES 31 & 32
N-(cis-2-(2-ethoxy-5,6-difluorophenyl)-cyclopropyl)-N'-(5-bromopyrid-2-yl)-
-urea
N-(trans-2-(2-ethoxy-5,6-difluorophenyl)-cyclopropyl)-N'-(5-bromopyrid-2-y-
l)-urea
[0241] The 2-ethoxy-5,6-difluorobenzaldehyde prepared as described
in Example 30 was converted to a cis/trans mixture of
2-(2-ethoxy-5,6-difluorophenyl)-cyclo-propylamines according to the
methods described in Examples 375 and 348 of WO 93/03022. This
product was then condensed with the product from Example 392
according to the procedure in Example 411 of WO 93/03022 and was
then oxidized with NBS as in Example 4 to provide a mixture of the
cis and trans
N-(2-(2-ethoxy-5,6-difluorophyl)-cyclopropyl)-N'-(5-bromopyrid-2-yl)-urea-
s.
[0242] The two products were separated on a silica-gel column
eluted with mixtures of dichloroethane and ethylacetate:
N-(cis-2-(2-ethoxy-5,6-diflu-
orophenyl)-cyclopropyl)-N'-(5-bromopyrid-2-yl)-urea: .sup.1H-NMR
(250 MHz, CDCl.sub.3): 1.2-1.3 (m, 1H), 1.35 (t, 3H), 1.5 (m, 1H),
2.0-2.1 (m, 1H), 3.2-3.3 (m, 1H), 3.95 (q, 2H), 6.5-6.6 (m, 1H),
6.7-6.8 (m, 1H), 7.0 (dd, 1H), 7.55 (dd, 1H), 7.8 (s, 1H), 8.7 (s,
1H), 9.2 (s, 1H).
[0243]
N-(trans-2-(2-ethoxy-5,6-difluoro-phenyl)-cyclopropyl)-N'-(5-bromop-
yrid-2-yl)-urea: .sup.1H-NMR (250 MHz, CDCl.sub.3): 1.2-1.3 (m,
1H), 1.4 (t, 3H), 1.6-1.7 (m, 1H), 2.1-2.2 (m, 1H), 3.2-3.3 (m,
1H), 4.05 (q, 2H), 6.5 (m, 1H), 6.7-6.8 (m, 1H), 7.6-7.8 (m, 2H),
8.2-8.4 (m, 1H), 9.0 (s, 1H), 9.3 (s, 1H).
EXAMPLES 33 & 34
N-(cis-2-(2-chloro-6-fluorophenyl)-cyclopropyl)-N'-(5-cyanopyrid-2-yl)-ure-
a
N-(cis-2-(2-chloro-6-fluorophenyl)-cyclopropyl)
-N'-(5-chloropyrid-2-yl) -urea
[0244] 2-chloro-6-fluorobenzaldehyde (Aldrich), was converted to
2-chloro-6-fluorostyrene according to the method described in
Example 377 of WO 93/03022 and then to
cis-2-(2-chloro-6-fluorophenyl) cyclopropylisocyanate analogously
to the method described in Example 348 of WO 93/03022. This
compound was then condensed with 2-amino-5-cyanopyridine of Example
41 in a manner analogous to the method in Example 7 to provide
N-(cis-2-(2-chloro-6-fluorophenyl)-cyclopropyl) -N'-(5-cya
nopyrid-2-yl)-urea: Mp 189.degree. C. .sup.1H-NMR (250 MHz,
DMSO-d.sub.6): 1.2 (m, 1H), 1.45-1.55 (m, 1H), 2.2-2.3 (m, 1H),
3.3-3.4 (m, 1H), 7.35-7.6 (m, 4H), 7.9-8.1 (m, 2H), 9.5 (s,
1H).
[0245] 2-(2-chloro-6-fluorophenyl)cyclopropylisocyanate prepared as
described above was condensed with 2-amino-5-chloropyridine
according to the method used in Example 7 to provide
N-(cis-2-(2-chloro-6-fluorophenyl-
)-cyclopropyl)-N'-(5-chloropyrid-2-yl)-urea: .sup.1H-NMR (250 MHz,
CDCl.sub.3): 1.3-1.4 (m, 1H), 1.5-1.6 (m, 1H), 2.0-2.1 (m, 1H),
3.3-3.4 (m, 1H), 6.7-6.8 (m, 1H), 6.9-7.0 (m, 1H), 7.15-7.30 (m,
2H), 7.4-7.5 (m, 1H), 7.7 (m, 1H), 8.8 (s, 1H), 9.1 (s, 1H)
EXAMPLE 35
(+,-)-N-(cis-2(2-Chloro-3-ethoxy-6-fluorophenyl)
cyclopropyl-N'-(5-acetopy- rid-2-yl)-urea
[0246] To a mixture of acetic acid (2 ml), H.sub.2SO.sub.4 (5
drops) and H.sub.2O (2 drops) was added
(+,-)-N-(cis-2(2-Chloro-3-ethoxy-6-fluorophe-
nyl)cyclopropyl-N'-(5-ethynpyrid-2-yl)-urea (40 mg, 0.11 mmole)
from Example 62. This mixture was stirred well and mercuric acetate
(50 mg, 0.16 mmole) added. After 1 hour, the mixture was
partitioned between aqueous KHCO.sub.3 and ethyl acetate. The
organic layer was dried, concentrated and the residue purified by
silica gel chromatography (toluene-acetone, 3:1) to yield the title
compound (11.2 mg, 27%). .sup.1H-NMR (250 MHz, CDCl.sub.3) d 1,35
(m, 1H), 1.50 (t, 3H), 1.64 (m, 1H), 2.13 (dd, 1H), 2.53 (s, 3H),
3.32 (m, 1H), 4.12 (dq, 2H), 6.81-6.96 (m, 3H), 8.03 (dd, 1H), 9.41
(broad s, NH), 9.62 (broad s, NH). .sup.13C-NMR (250 MHz,
CDCl.sub.3) selected signals d 14 81, 15.12, 15.80, 26.21, 65.57,
111.46, 126.03, 137.62, 147.73, 156.88, 156.73, 195.29.
EXAMPLE 36
N-(2-(Imidazo-4-yl)ethyl)-N'-(5-bromopyrid-2-yl)-urea
[0247] To a stirred solution of
N-(2-(imidazo-4-yl)ethyl)-N'-(5-bromopyrid- -2-yl)-thiourea
prepared analogously to Example 328 of WO 93/03022 (50 mg, 0.15
mmole) in dioxane (7 ml) and N.sub.2O (1 ml) was added
N-bromosuccinimide (100 mg, 0.56 mmole). After 1 hr. the mixture
was partitioned between aqueous NaHCO.sub.3 and
CH.sub.2Cl.sub.2.
[0248] The organic layer was dried, concentrated and crystallised
from ethanol to give the title compound (22 mg, 46%). MP:
204.degree. C. .sup.1H-NMR (250 MHz, CDCl.sub.3) d 3.05 (m, 2H),
3.60 (m, 2H), 6.87 (d, 1H), 7.03 (dd, 1H), 7.27 (s, 1H), 7.71 (dd,
1H), 8.29 (d, 1H), 8.39 (d, 1H), 10.48 (broad s, 1H). .sup.13C-NMR
(250 MHz, CDCl.sub.3) d 20.32, 38.82, 112.99, 123.18, 125.70,
134.64, 140.44, 145.94, 146.59, 159.85.
EXAMPLE 37
N-[2-(2,6-Difluoro-3-dimethylamino)phenethyl]-N'-[2-(5-chloro)pyridyl]-ure-
a
[0249] A mixture of 2,4-difluoroaniline (5.0 g, 38.7 mmole) and
trimethylphosphate (3.6 g, 25.8 mmole) was refluxed at 180.degree.
C. for 2 hrs. After the mixture was cooled to 50.degree. C. NaOH
(3.2 g, 80.6 mmole) in 12 ml of H.sub.2O was added to it and it was
refluxed again for 1 hr. The mixture was cooled, H.sub.2O was added
to it and it was extracted with diethyl ether, dried over
Na.sub.2SO.sub.4 and evaporated. The crude material was filtrated
through Al.sub.2O.sub.3 column by using diethyl ether as eluent to
give 4.4 g (73%) of 1-dimethylamino-2,4-difluo- robenzene. This
compound was converted to 2,6-difluoro-3-dimethylamino-phe-
nethylamine according to the procedure in Examples 362 and 151 of
WO 93/03022, and condensed with the product from Example 392
according to the procedure in Example 411 to give
N-[2-(2,6-difluoro-3-dimethylamino)p- henethyl]-N'[2-(5-chl
oro)pyridyl]-thiourea. This compound was converted to the titled
compound according to the procedure in the present Example 4. Mp
135.degree. C. .sup.1H NMR (250 MHz, CDCl.sub.3) .delta.9.64 (s,
1H), 9.21 (br s, 1H), 8.13 (d, 1H), 7.65 (dd, 1H), 6.86-6.75 (m,
3H), 3.63 (q, 2H), 3.01 (t, 2H), 2.75 (s, 6H).
EXAMPLE 38
N-(2-Phenethyl)-N'-[2-(5-chloro)pyridazyl]-urea
[0250] NaH, 80% (0.12 g, 3.86 mmole) was added to a solution of
3-amino-6-chloropyridazine (0.50 g, 3.86 mmole) in THF (40 mL) at
0.degree. C. under nitrogen. The mixture was stirred for 15 min and
N,N'-carbonyldiimidazole (0.63 g, 3.86 mmole) in 40 mL of THF
added. The temperature of the mixture was allowed to rise to room
temperature and it was stirred for 3 hrs. at this temperature,
refluxed for 3 hrs., phenethylamine (0.51 mL, 3.86 mmole) was added
and the reflux continued for 1 hr. Diethyl ether and NH.sub.4Cl
(aq) was added to the mixture and the organic phase separated and
washed with brine, dried over Na.sub.2SO.sub.4 and evaporated. The
residue was purified by chromatography on silica gel column to
yield the titled compound. Mp 204.degree. C. .sup.1H NMR (250 MHz,
CDCl.sub.3) .delta.11.32 (br s, 1H), 8.29 (d, 1H), 7.48 (d, 1H),
7.28 (m, 6H), 3.59 (t, 2H), 2.96 (t, 2H).
EXAMPLE 39
(+,-)-N-[cis-2-(6-Chloro-3-ethoxy-2-fluorophenyl)
cyclopropyl]-N'-[2-(5-ch- loro)pyridazyl]-urea
[0251] The titled compound was obtained starting from
4-chloro-2-fluorophenol which was converted to
6-chloro-3-ethoxy-2-fluoro- benzaldehyde according to procedure in
Example 362 of WO 93/03022. This aldehyde was converted to
6-chloro-3-ethoxy-2-fluorostyrene according to procedure in Example
375 of WO 93/03022. Styrene (30.7 g, 153 mmole) was dissolved in
dichloroethane (400 mL) and CuJ (50 mg) and Pd(OAc).sub.2 (50 mg)
added. The temperature of this solution was raised to reflux and
diethyl azodicarboxylate (32.2 mL, 306 mmole) in 100 mL of
dichloroethane was dropped into it during a period of 2 hrs. The
reflux was continued for 1 hr. The reaction mixture was evaporated
and the residue purified by chromatography on silica gel column
EtOAc/Hexane 2:98-5:95 as eluent to give 4.36 g (10%) of pure
(+,-)-cis-2-(6-chloro-3-ethoxy-2-fluorophenyl)--
cyclopropylcarboxylic acid ethyl ester which was then hydrolyzed in
KOH (2.56 g)--EtOH/H.sub.2O (45 mL/5 mL) at 100.degree. C. for 24
hrs. The reaction mixture was extracted twice with hexane, made
acidic with conc. HCl and extracted with diethyl ether which was
dried over Na.sub.2SO.sub.4 and evaporated to give 3.87 g (98%) of
pure acid.
[0252]
(+,-)-cis-2-(6-Chloro-3-ethoxy-2-fluorophenyl)-cyclopropylcarboxyli-
c acid (1.0 g, 3.87 mmole), diphenyl phosphoryl azide (0.92 mL,
4.26 mmole) and triethylamine (0.59 mL, 4.26 mmole) in 25 mL of
toluene was refluxed for 40 min. 3-Amino-6-chloropyridazine (0.55
g, 4.26 mmole) in 20 mL of DMF was added and reflux was continued
for 3 hrs. The toluene was evaporated. H.sub.2O was added to the
residue and extracted with diethyl ether. The precipitate which
formed in the H.sub.2O phase (a part of the product) was filtrated,
the diethyl ether phase was dried over Na.sub.2SO.sub.4 and
evaporated (the other part of the product). The collected crude
material (1.2 g) was recrystallized from acetonitrile to give 0.7 g
(47%) of the titled compound. Mp 210-211.degree. C. .sup.1H NMR
(250 MHz, DMSO-d6) .delta.9.87 (s, 1H), 8.10 (d, 1H), 7.81 (d, 1H),
7.33 (d, 1H), 7.17 (t, 1H), 7.01 (br s, 1H), 4.16 (q, 2H), 3.33 (m,
1H), 2.13 (q, 1H), 1.58 (q, 1H), 1.41 (t, 3H), 1.22 (m, 1H).
EXAMPLE 40
(+,-)-N-[cis-2-(2-Chloro-3-ethoxy-6-fluorophenyl)
cyclopropyl]-N'-[2-(5-ch- loro)pyridazyl]-urea
[0253] The titled compound was obtained starting from
2-chloro-4-fluorophenol and prepared as described in Example 39. Mp
192.degree. C. .sup.1H NMR (250 MHz, CDCl.sub.3) .delta.10.70 (br
s, 1H), 8.00 (m, 1H), 7.37 (d, 1H), 6.86 (t, 1H), 6.74 (m, 1H),
4.05 (q, 2H), 3.44 (m, 1H), 2.16 (q, 1H), 1.64 (q, 1H), 1.43 (m,
4H).
EXAMPLE 41
(+,-)-N-[cis-2-(2-Chloro-3-ethoxy-6-fluorophenyl)
cyclopropyl]-N'-[2-(5-cy- ano)pyridyl]-urea
[0254] Preparation of 2-amino-5-cyanopyridine: A mixture of
NiBr.sub.2 (2.18 g, 10 mmole), triphenylphosphine (10 g, 38 mmole)
and zinc powder (1 g, 15.2 mmole) in 100 mL of acetonitrile was
stirred under nitrogen at 60.degree. C. for 1 hr. NaCN (6.3 g, 102
mmole) and 2-amino-5-bromopyridine (17.8 g, 100 mmole) were added
to the mixture which was stirred overnight at 60.degree. C. 500 mL
of ethyl acetate was added to the mixture and it was filtered and
the solvent evaporated. The product was purified on a silica gel
column eluted with ethyl acetate. .sup.1H NMR (250 MHz,
DMSO-d.sub.6) .delta.8.41 (d, 1H), 7.29 (dd, 1H), 7.12 (s, 2H),
6.58 (d, 1H).
[0255] The titled compound was obtained starting from
2-chloro-4-fluorophenol and prepared as described in Example 39.
The last step was made using 2-amino-5-cyanopyridine prepared above
instead of 2-amino-5-chloropyridazine. Mp 201.degree. C. .sup.1H
NMR (250 MHz, CDCl.sub.3) .delta.9.83 (br s, 1H), 9.36 (br s, 1H),
8.14 (d, 1H), 7.71 (dd, 1H), 6.93-6.84 (m, 3H), 4.11 (q, 2H), 3.29
(m, 1H), 2.14 (q, 1H), 1,64 (m, 1H), 1.53 (t, 3H), 1.33 (m,
1H).
EXAMPLE 42
(+,-)-N-[cis-2-(2-Chloro-3-ethoxy-6-fluorophenyl)
cyclopropyl]-N'[5-(4-bro- mo-3-methyl)pyrazyl]-urea
[0256] The titled compound was obtained starting from
2-chloro-4-fluorophenol and prepared as described in Example 39.
The last step was made using 5-amino-4-bromo-3-methylpyrazole
hydrobromide instead of 2-amino-5-chloropyridazine. .sup.1H NMR
(250 MHz, CDCl.sub.3) .delta.7.30 (m, 1H), 7.01 (br s, 1H),
6.96-6.80 (m, 2H), 5.40 (br s, 1H), 4.07 (q, 2H), 3.30 (m, 1H),
2.14 (m, 1H), 2.00 (s, 3H), 1.65 (q, 1H), 1.44 (m, 4H).
EXAMPLE 43
(+,-)-N-[cis-2-(6-chloro-3-ethoxy-2-fluorophenyl)
cyclopropyl]-N'[2-(5-cya- no)pyridyl]-urea
[0257] The titled compound was obtained starting from
4-chloro-2-fluorophenol and prepared as described in Examples
39+41. Mp 211-212.degree. C. .sup.1H NMR (250 MHz, DMSO-d.sub.6)
.delta.9.94 (s, 1H), 8.43 (s, 1H), 8.15 (d, 1H), 8.03 (br s, 1H),
7.53 (d, 1H), 7.40 (d, 1H), 7.19 (t, 1H), 4.16 (q, 2H), 3.30 (m,
1H), 2.16 (q, 1H), 1.42 (t, 3H), 1.19 (m, 1H).
EXAMPLE 44
N-[2-(3-Acetyl-6-fluoro-2-methoxy)phenethyl]-N'-[2-(5-chloro)pyridyl]-urea
[0258]
N-[2-(3-Acetyl-6-fluoro-2-methoxy)phenethyl]-N'-[2-(5-chloro)pyridy-
l]-thiourea from Example 55 (1.0 g, 2.6 mmole) was dissolved in 20
mL of dioxane/H.sub.2O (10:1) and AgNO.sub.3 dissolved in 5 mL of
H.sub.2O was added dropwise to this solution at room temperature.
The mixture was stirred for 2 hrs. at room temperature. EtOAc was
added to this mixture and it was filtrated through a Celite column
EtOAc as eluent. The filtrate was was washed with H.sub.2O and
brine, dried over NaSO.sub.4 and evaporated. The residue was
purified by Silicagel column chromatography EtOAc/hexane
50:50--EtOAc 100 as eluent to give the titled compound. Mp
180.degree. C. .sup.1H NMR (250 MHz, CDCl.sub.3) .delta.9.75 (m,
1H), 8.80 (br s, 1H), 7.63 (d, 1H), 7.54 (t, 1H), 7.10 (d, 1H),
6.88 (t, 1H), 3.80 (s, 3H), 3.58 (q, 2H), 3.01 (t, 2H), 2.57 (s,
3H).
EXAMPLE 45
N-(2-phenethyl)-N'-(2-benzothiazolyl)-urea
[0259] 2-phenethylamine (0.88 ml, 7 mmolee) was added dropwise into
the solution of 1,1'-carbonyldimidazole (1.24 g, 7.7 mmolee) in
dichloromethane (21 ml) at 0.degree. C. After 3 hrs, the reaction
mixture was evaporated in vacuo; and the residue was redissolved in
dimethylformaldehyde (20 ml). To the solution was added
2-aminobenzothiazole (1.05 g, 7 mmolee). The reaction was kept
under 100.degree. C. overnight. It was the poured into water (100
ml) and extracted with dichloromethane twice.
[0260] The organic phase was washed succesively with 0.5 N
hydrochloric acid solution and water, and subsequently dried in
vacuo. The product was crystallized from dichloromethane (1.16 g).
Mp 136.degree. C. .sup.1H NMU (CDCl.sub.3): 7.22 (m, 9H, phenyl and
benzo), 3.60 (q, 2H, CH.sub.2NH), 2.75 (t, 2H, CH.sub.2).
EXAMPLE 46
N-(2-phenethyl)-N', N'-methyl-(2-thiazolyl)-urea
[0261] Acetic anhydride (10 ml) and formic acid (10 ml) were mixed
at 0.degree. C., and then heated to 50.degree. C. for 15 min. After
cooling to 0.degree. C., a mixture of 2-aminothiazole (2 g, 20
mmolee) and formic acid (20 ml) was added to it. The reaction was
kept at room temperature overnight. After evaporation and
coevaporation with toluene, 2-formamidothiazole was obtained (2.2
g).
[0262] 2-formamidothiazole (1.6 g, 14 mmolee) was dissolved in dry
tetrahydrofuran. To the solution was added aluminium hydride (0.8
g, 21 mmolee). The reaction mixture was refluxed for 3 hrs, and
poured onto ice. It was extracted with dichloromethane three times,
and the organic solution was washed with water and evaporated.
2-methylaminothiazole was isolated by alumina column
chromatography.
[0263] 2-phenethylamine (0.38 ml, 3 mmolee) was added dropwise into
the solution of 1,1'-carbonyldimidazole (0.48 g, 3 mmolee) in
dichloromethane (15 ml) at 0.degree. C. After 1 hr., the reaction
mixture was evaporated in vacuo; and the residue was redissolved in
dimethylformaldehyde (10 ml). To the solution was added
2-methylaminothiazole (0.25 g, 2 mmolee). The reaction was kept
under 100.degree. C. for two hours. It was then poured into water
(100 ml) and extracted with dichloromethane twice. The organic
phase was washed successively with 0.5 N hydrochloric acid solution
and water, and subsequently dried in vacuo. The title product was
isolated by silica gel column chromatography (203 mg). .sup.1H NMR
(CDCl.sub.3: 7.27 (m, 6H, phenyl and thiazole), 6.86 (d, 1H,
thiazole), 3.63 (q, 2H, CH.sub.2NH), 3.43 (s, 3H, CH.sub.3), 2.90
(t, 2H, CH.sub.2).
EXAMPLES 47 & 48
N-(cis)
-2-(2,5-Dimethoxyphenyl)-cyclopropyl-N'-(5-chloropyrid-2-yl)-urea
N-(trans)
-2-(2,5-Dimethoxylphenyl)-cyclopropyl-N'-(5-chloropyrid-2-yl)-ur-
ea
[0264] 2,5-Dimethoxybenzaldehyde (Aldrich) was converted to a
cis/trans mixture of 2-(2,5-dimethoxyphenyl)-cyclopropylisocyanates
according to the methods described in Examples 375 and 348 of WO
93/03022. This product was then condensed with
2-amino-5-chloropyridine in a manner analogous to the method in
Example 7 to provide cis/trans mixture of
N-2-(2,5-Dimethoxyphenyl)-cyclopropyl-N'-(5-chloro-pyrid-2-yl)-ureas.
The two products were separated on a silica-gel column eluated with
mixtures of hexane and ethylacetate:
N-(cis)-2-(2,5-Dimethoxy-phenyl)cyclopropyl-N-
'-(5-chloro-pyrid-2-yl)-urea: Mp 155-157.degree. C. .sup.1H NMR
(250 MHz, CDCl.sub.3) .delta.9.81 (br s, 1H), 8.85 (br s, 1H), 7.57
(s, 1H), 7.40 (dd, 1H), 6.92-6.63 (m, 4H), 3.79 (s, 3H), 3.70 (s,
3H), 3.29 (m, 1H), 2.41 (q, 1H), 1.39-0.97 (m, 2H).
[0265] N-(trans)
-2-(2,5-Dimethoxylphenyl)-cyclopropyl-N'-(5-chloropyrid-2-
-yl)-urea: .sup.1H NMR (250 MHz, CDCl.sub.3) .delta.9.45 (br s,
1H), 8.23 (s, 1H), 7.62 (d, 1H), 6.89-6.62 (m, 3H), 6.59 (m, 1H),
3.82 (s, 3H), 3.77 (s, 3H), 3.08-2.86 (m, 1H), 2.42-2.29 (m, 1H),
1.39-1.20 (m, 2H).
EXAMPLE 49
N-(cis)
-2-(2,5-Dimethoxyphenyl)-cyclopropyl-N'-(5-bromopyrid-2-yl)-urea
[0266] 2-(2,5-Dimethoxyphenyl)-cyclopropylisocyanates prepared as
described in the Example above, were condensed with
2-amino-5-bromopyridine according to the method used in Example 7
to provide
N-(cis)-2-(2,5-Dimethoxyphenyl)-cyclopropyl-N'-(5-bromopyrid-2-yl-
)-urea after chromatography on silica gel. Mp 191-192.degree. C.
.sup.1H NMR (250 MHz, CDCl.sub.3) .delta.8.79 (br s, 1H), 7.70 (br
s, 1H), 7.55 (dd, 1H), 6.88-6.62 (m, 4H), 3.81 (s, 3H), 3.72 (s,
3H), 3.29 (m, 2H), 2.40 (q, 2H), 1.41-1.30 (m, 1H), 1.09-0.98 (m,
1H).
EXAMPLE 50
N-(cis)
-(3-Ethoxy-2-fluoro-6-methoxyphenyl)-cyclopropyl-N'-(5-bromopyrid--
2-yl)-urea
[0267] A suspension of 2-fluoro-4-methyl acetophenone (10.0 g,
0.059 mol), mCPBA (26.0 g, 0.128 mol) and MgSO.sub.4 (30.0 g) in
500 mL of CHCl.sub.3 was stirred for 12 hrs. at RT. The mixture was
then filtrated, washed with 2M NaOH, dried over Na.sub.2SO.sub.4
and concentrated in vacuo to give
1-acetyl-2-fluoro-4-methoxybenzene as a solid (10.7 g, 98%). A
solution of the 1-acetyl-2-fluoro-4-methoxybenzene (5.3 g, 0.029
mol) in 100 mL of MeOH and 20 mL of 25% NH.sub.3 (aq) was stirred
for 1 hr. at RT. The mixture was then concentrated in vacuo,
diluted with water, extracted with EtOAc, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo to give
2-fluoro-4-methoxyphenol as an oil (3.85 g, 94%).
2-Ethoxy-1-fluoro-4-methoxybenzaldehyde was prepared according to
method described in Example 362 of WO 93/03022 and converted to a
cis/trans mixture of
2-(2-ethoxy-1-fluoro-6-methoxyphenyl)cyclopropylisocyanates
according to the methods described in Examples 375 and 348 of WO
93/03022.
[0268] This product was then condensed with
2-amino-5-bromo-pyridine in a manner analogous to the method in
Example 7 to provide cis/trans mixture of
N-2-(2-ethoxy-1-fluoro-6-methoxyphenyl)cyclopropyl-N'-(5-bromopyrid-2--
yl)-ureas. The two products were separated on a silica-gel column
eluated with mixtures of hexane and ethylacetate to yield
N-(cis)-(3-ethoxy-2-flu-
oro-6-methoxyphenyl)-cyclopropyl-N'-(5-bromopyrid-2-yl)-urea:
.sup.1H NMR (250 MHz, CDCl.sub.3) .delta.7.88 (s, 1H), 7.55 (dd,
1H), 6.83 (t, 1H), 6.58 (dd, 1H), 4.03 (q, 2H), 3.79 (s, 3H),
3.32-3.19 (m, 1H), 1.54-1.20 (m, 2H), 1.46 (t, 3H).
EXAMPLE 51
N-(3-Ethoxy-2-fluoro-6-methoxyphenetyl)-N'-(5-chloropyrid-2-yl)-urea
[0269] 3-Ethoxy-2-fluoro-6-methoxyphenethylamine was prepared from
2-ethoxy-1-fluoro-4-methoxy-benzaldehyde (prepared as described in
the Example above) as described in Example 151 of WO 93/03022.
3-Ethoxy-2-fluoro-6-methoxyphenethylamine was reacted with
5-chloropyrid-2-ylisothiocyanate as described in Example 370 of WO
93/03022 to give
N-[2-(3-ethoxy-2-fluoro-6-methoxy)-phenethyl]-N'-[2-(5-c-
hloro)pyridyl]-thiourea. The title compound was obtained from
N-[2-(3-ethoxy-2-fluoro-6-methoxy)phenethyl]-N'-[2-(5-chloro)pyridyl]-thi-
ourea according to the procedure in Example 4. Mp 194-196.degree.
C.; .sup.1H NMR (250 MHz, CDCl.sub.3) .delta.9.20 (br s, 1H), 9.05
(br s, 1H), 8.05 (d, 1H), 7.50 (dd, 1H), 6.82 (d, 1H), 6.74 (d,
1H), 4.00 (q, 2H), 3.73 (s, 3H), 3.59 (dd, 2H), 2.98 (dd, 2H), 1.41
(t, 3H).
EXAMPLE 52
N-(cis)
-2-(2-Chloro-3-ethoxy-6-fluorophenyl)-cyclopropyl-N'-(6-bromo-5-hy-
droxypyrid-2-yl)-urea
[0270] Starting materials 3-acetyl-6-amino-2-bromopyridine and
2-(2-chloro-3-ethoxy-6-fluorophenyl)-cyclopropylisocyanate, were
prepared in following manner: 5 g of 2-Bromo-3-pyridinol (Aldrich)
was added to a mixture of 25 ml of conc. H.sub.2SO.sub.4 and 25 ml
of fuming HNO.sub.3 at 0.degree. C. The mixture was stirred one
hour at that temperature, then added to 350 ml of crushed ice and
extracted with CH.sub.2Cl.sub.2. The extract was dried with
Na.sub.2SO.sub.4 and evaporated in vacuo to yield 4.1 g of a crude
product as a mixture of 2-bromo-6-nitro-3-pyridino- l and
2-bromo-4,6-dinitro-3-pyridinol. The mixture (0.5 g) was treated
with an excess of acetic anhydride at room temperature at 24 hours,
then separated on a silica-gel column eluated with mixtures of
hexane and ethylacetate to yield 0.28 g of
2-bromo-6-nitro-3-pyridinol as a yellowish solid.
2-Bromo-6-nitro-3-pyridinol was then treated with acetic anhydride
at 80.degree. C. at 14 hours under an atmosphere of nitrogen to
yield 0.34 g of 3-acetyl-2-bromo-6-nitropyridine as a yellow solid.
Hydrogenation of 3-acetyl-2-bromo-6-nitropyridine in ethanol (Pt on
C, 1 atm) gave 3-acetyl-6-amino-2-bromopyridine as a reddish oil.
The cis/trans mixture of
2-(2-chloro-3-ethoxy-6-fluorophenyl)cyclopropyliso-c- yanates were
prepared from 2-chloro-4-fluorophenol (Aldrich) in manner analogous
to Examples 362, 375 and 348 of WO 93/03022. This product was then
condensed with 3-acetyl-6-amino-2-bromopyridine in a manner
analogous to the method in Example 7 to provide cis/trans mixture
of
N-2-(2-Chloro-3-ethoxy-6-fluorophenyl)-cyclopropyl-N'-(6-bromo-5-hydroxyp-
yrid-2-yl)-ureas. The two products were separated on a silica-gel
column eluated with mixtures of hexane and ethylacetate to yield
the titled compound as a white solid: .sup.1H NMR (250 MHz,
CDCl.sub.3) .delta.7.85 (d, 1H), 7.55 (br s, 1H), 6.98-6.72 (m,
2H), 4.04 (q, 2H), 3.27-3.04 (m, 1H), 2.18 (q, 1H), 1.74-1.65 H (m,
1H), 1.43 (t, 3H), 0.92-0.80 (m, 2H).
EXAMPLE 53
N-(cis/trans)-2-(2-Chloro-3-ethyl-6-fluorophenyl)-cyclopropyl-N'-(6-bromo--
5-methoxypyrid-2-yl)-urea
[0271] Starting material -2-amino-6-bromo-5-methoxybromo-pyridine,
was prepared in following manner: 2-bromopyridin-3-ol (Aldrich) was
converted to 2-bromo-3-methoxy-6-nitropyridine as described in J.
Med. Chem. 1981, 24, 39-42. Hydrogenation of
2-bromo-6-nitro-5-methoxypyridine in ethanol (Pt on C, 1 atm) gave
the starting material as a reddish oil. This product was then
condensed with cis/trans mixture of
2-(2-chloro-3-ethoxy-6-fluorophenyl)cyclopropyliso-cyanates,
described in Example 53, in a manner analogous to the method in
Example 7 to provide cis/trans (50:50) mixture of
N-2-(2-Chloro-3-ethoxy-6-fluorophenyl)-cyclo-
propyl-N'-(6-bromo-5-hydroxypyrid-2-yl)-ureas. .sup.1H NMR (250
MHz, CDCl.sub.3) .delta.7.62-7.43 (m, 1H), 7.29-7.12 (m, 1H),
6.91-6.67 (m, 2H), 4.10-3.94 (m, 2H), 3.90 and 3.84 (2 s,
3H),3.38-3.02 (m, 1H) 2.18-2.00 (m, 1H), 1.55-1.30 H (m, 5H), 1.43
(t, 3H), 0.92-0.80 (m, 2H).
EXAMPLE 54
N-(3-Acetyl-2-fluoro-6-methoxyphenetyl)-N'-(5-chloropyrid-2-yl)-urea
[0272] Starting material 3-acetyl-2-fluoro-6-methoxyphenetyl-amine,
was prepared in following manner: A mixture of
2-Fluoro-4-methoxyacetophenone (5 g)(Fluorochem), ethylene glycol
(4 ml), and p-toluensulfonic acid (50 mg) in toluene was refluxed
10 hours to give 5.7 g of
2-(2-fluoro-4-methoxyphenyl)-2-methyl-1,3-dioxolane. This was
converted to
2-fluoro-6-methoxy-3-(2-methyl-1,3-dioxol-2-yl)-benzaldehyde
according to Example 362 of WO 93/03022. This compound was
converted to 2-fluoro-6-methoxy-3-(2-methyl-1,
3-dioxol-2-yl)-cinnamic acid ethylester according to the methods
described in Example 353 of WO 93/03022.
2-Fluoro-6-methoxy-3-(2-methyl-1,3-dioxol-2-yl)cinnamic acid
ethylester was then treated with KOH in dioxane and water to yield
2-fluoro-6-methoxy-3-(2-methyl-1,3-dioxol-2-yl)cinnamic acid.
Hydrolysis with aqueous HCl in dioxane gave
3-acetyl-2-fluoro-6-methoxycinnamic acid.
N-(3-Acetyl-2-fluoro-6-methoxyphenetyl)-N'-(5-chloropyrid-2-yl)-ure-
a was then prepared in a manner analogous to the method in Example
7. .sup.1H NMR (250 MHz, CDCl.sub.3) .delta.9.18 (br s, 1H), 8.97
(br s, 1H), 7.85 (d, 1H), 7.83 (t, 1H), 7.51 (dd, 1H), 6.83 (d,
1H), 6.70 (d, 1H), 3.82 (s, 3H), 3.57 (q, 2H), 3.08-2.92 (m, 2H),
2.51 (d, 3H).
EXAMPLE 55
(+,-)-N-(cis-2-(2-fluoro-5-butoxyvinyl-6-methoxymethoxyph
enyl)cyclopropyl)-N'-(5-chloropyrid-2-yl)-urea
[0273] 3-Fluorophenol (40 ml, 442 mmole) and 2,3-dihydropyrane (45
ml, 500 mmole) were dissolved in dichloromethane and hydrogen
chloride in ether (20 ml, 1M) was added. The solution was stirred
for 2 hours and was washed with sodium hydroxide in water followed
by water.
[0274] The solution was dried with sodium sulphate, filtered and
evaporated. The residue was distilled at 0.5 mm Hg at 85.degree. C.
to furnish 50 g of collarless oil. The oil was dissolved in THF
under nitrogen and the solution was cooled to -78.degree. C.
n-Butyllithium (112 ml, 2,5M) was added.
[0275] The solution was stirred at -78.degree. C. for 40 minutes.
Triethylorthoformate (47 ml, 282 mmole) was added followed by
trimethylsilyltriflate (54 ml, 280 mmole). The solution was diluted
with hexane and washed with water. The organic phase was dried with
sodium sulphate, filtered and evaporated. The residue was purified
on silica eluting with ethyl acetate-hexane 1:10. The pure
fractions were combined and evaporated to yield 30 g of an oil. The
oil was dissolved in THF under nitrogen. The solution was cooled to
-78.degree. C. and n-BuLi (44 ml, 2.5 M) was added. The solution
was warmed to -30.degree. C. for 20 seconds and was then cooled to
-50.degree. C. for 5 minutes and then further cooled to -78.degree.
C. for 30 minutes. Iodine (28.1 g, 111 mmole) dissolved in THF was
added. The solution was diluted with hexane and washed with water
containing a little sodium thiosulphate. The solution was dried
with sodium sulphate, filtered and evaporated. The residue was
purified by recrystallisation from n-hexane to yield 23.22 g of
solid. The solid was dissolved in dioxane and dilute hydrochloric
acid was added. The solution was stirred for 60 minutes, diluted
with hexane and washed with water. The organic phase was dried with
sodium sulphate, filtered and evaporated to yield 13.5 g of solid.
The solid was dissolved in dichloromethane and triethylamine (8 ml)
and methoxymethylchloride (4.1 ml) was added. The solution was
stirred for 60 minutes and was washed with water dried with sodium
sulphate, filtered and evaporated. The residue was added to a
pre-formed solution of ethylenetriphenyl-phosp- horane (56 mmole)
in THF under nitrogen. The solution was stirred for 60 minutes at
60.degree. C. The solution was diluted with hexane, washed with
water, dried with sodium sulphate, filtered and evaporated. The
residue was purified on silica by elution with ethyl acetate-hexane
1:20. The pure fractions were collected and evaporated to yield 13
g of an oil. The oil was dissolved in dichloroethane and
copper(1)triflate (20 mg) and palladium(2)acetate (20 mg was added)
the solution was heated to reflux and ethyldiazoacetate (50 ml) was
added slowly. The solution was washed with water and evaporated.
The residue was purified on silica by elution with ethyl
acetate-hexane 1:9. The purest fractions containing the cis-isomere
were collected and evaporated. The residue was dissolved in
methanol and sodium hydroxide in water (ca 15%) was added. The
solution was stirred for 60 minutes at 40.degree. C. This treatment
hydrolyzed the remaining trans-isomer as well as some other
impurity's, but not the wanted cis-ester. The solution was diluted
with water and ether, the organic phase washed with water, dried
with sodium sulphate, filtered and evaporated to yield 3.3 g pure
cis-isomer as an oil. The oil was dissolved in DMF and the solution
was degassed 5 mm Hg. Tallium acetate (2.5 g) and n-butylvinylether
(5.6 ml) and bis-diphenylphosphinopropane (0.2 g) and palladium
acetate (0.1 g) was added under nitrogene. The solution was heated
to 90.degree. C. overnight. The solution was diluted with ether,
washed with water, dried with sodium sulphate, filtered and
evaporated. The residue was purified by on silica by elution with
ethyl acetate-hexane 1:9. The pure fractions were collected and
evaporated to yield 1.8 g of an oil. The oil was dissolved in
methanol and potassium hydroxide (370 mg) was added. The solution
was refluxed overnight. The solution was evaporated and
co-evaporated 3 times with THF and the solid residue was put onto a
freeze drier overnight. 0.5 g of the solid was dissolved in toluene
and diphenylphosphorylazide (1 eq.) was added. The solution was
refluxed for 20 minutes. 2-Amino-5-chloropyridine (1.2 eq.) was
added and the reflux was continued for 40 minutes. The solution was
diluted with ethylacetate washed with water, dried with sodium
sulphate, filtered and evaporated. The residue was purified on
silica by elution with ethylacetate-hexane 1:2. The pure fractions
were collected and evaporated to yield 127 mg of the title product.
.sup.1H-NMR CDCl.sub.3 .delta.0.9 (t, 3H) 1.3-1.8 (m, 6H) 2.1 (q,
1H) 3.3-3.4 (bs, 1H) 3.5 (s, 3H) 3.8 (t, 2H) 4.3-4.5 (m, 2H) 5.1
(s, 2H) 6.8 (m, 2H) 7.3-7.5 (m, 2H) 7.8 (s, 1H) 9.1 (bs, 1H) 9.8
(bs, 1H).
EXAMPLE 56
(+,-)-N-(cis-2-(5-Acetyl-2-fluoro-6-methoxymethoxy-phenyl)cyclopropyl)-N'--
(5-chloropyrid-2-yl)-urea
[0276]
(+,-)-N-(cis-2-(2-fluoro-5-butoxyvinyl-6-methoxymethoxy)-cyclopropy-
l)-N'-(5-chloropyrid-2-yl)-urea (Example 55) was dissolved in
dioxane and a few drops of dilute hydrochloric acid was added. The
solution was stirred for 15 minutes, was diluted water and
evaporated.
[0277] The residue was co-evaporated with water to yield the title
product as a solid. .sup.1H-NMR CDCl.sub.3 1.3 (q, 1H) 1.5 (q, 1H)
2.1 (q, 1H) 2.5 (s, 3H) 3.4 (m, 1H) 3.5 (s, 3H) 5.1 (m, 2H) 6.8 (m,
2H) 7.4-7.6 (m, 2H) 7.8 (d, 1H) 9.1 (bs, 1H) 9.8 (bs, 1H).
EXAMPLE 57
(+,-)-N-(cis-2-(5-Acetyl-2-fluoro-6-hydroxyphenyl)-cyclopropyl)-N'-(5-chlo-
ropyrid-2-yl)-urea
[0278]
(+,-)-N-(cis-2-(5-Acetyl-2-fluoro-6-methoxymethoxy)-cyclopropyl)-N'-
-(5-chloropyrid-2-yl)-urea (Example 56) was dissolved in dioxane
and concentrated hydrochloric acid was added. The solution was
stirred for 15 minutes, was diluted water and evaporated. The
residue was co-evaporated with water to yield the title product as
a solid. .sup.1H-NMR DMSO-d.sub.6 .delta.1.2 (m, 1H) 1.3-1.5 (m,
1H) 2.0 (q, 1H) 2.6 (s, 3H) 3.4 (m, 1H) 6.8 (t, 1H) 7.4 (d, 1H) 7.7
(m, 1H) 7.8-8.0 (m, 2H) 9.2 (s, 1H) 11.1 (bs, 1H).
EXAMPLE 58
N-2-(5-Acetyl-2-fluoro-6-hydroxyphenyl)-ethyl)-N'-(5-chloropyrid-2-yl)-ure-
a
[0279] 4-Fluoro-2-hydroxiacetophenone, (CA registry number:
1481-27-2) (43 g, 311 mmole) was dissolved in dry THF under
nitrogen and benzylbromide (35 ml, 295 mmole) was added followed by
sodium hydride, 60% in oil (13 g, 311 mmole). The solution was
stirred at 50.degree. C. for 5 hours, diluted with hexane and
washed with water. The organic phase was dried with sodium
sulphate, filtered and evaporated. The residue was dissolved in a
mixture of ethylenglycol (100 ml), dioxane (100 ml) and
triethylorthoformate (40 ml). Hydrogen chloride in ether (30 ml,
1M) was added and the solution was stirred for 2 hours The solution
was diluted with hexane, washed with water dried with sodium
sulphate, filtered and evaporated. The residue was distilled at
140.degree. C. and 5 mm Hg to yield 49 g of an oil. 43 g of the oil
was dissolved in dry THF under nitrogen and was cooled to
-78.degree. C. and n-BuLi (43 ml, 2,5M) was added slowly. The
solution was stirred for 40 minutes and DMF (30 ml) was added. The
solution was diluted with ethyl acetate-water and the organic phase
was washed with water, dried with sodium sulphate, filtered and
evaporated. The residue was crystallised from hexane to yield 28 g
of solid. The solid was dissolved in THF and sodium hydride (5 g,
60% in oil) followed by nitromethane (20 ml) was added. The
solution was stirred for 2 hours at 45.degree. C. The solution was
diluted with ether, washed with water, dried with sodium sulphate,
filtered and evaporated to yield 37 g of an oil.
[0280] The oil was dissolved in dichloromethane and the solution
was cooled to 0.degree. C. Triethylamine (30 ml, 216 mmole) and
methanesulphonyl-chloride (9.4 ml, 96 mmole) was added. The
solution was stirred at 0.degree. C. for 30 minutes and was then
washed with water dried with sodium sulphate, filtered and
evaporated to yield 31 g of a solid. The solid was dissolved in THF
and LAH (15 g) was added slowly. The solution was refluxed for 30
minutes.
[0281] Water (15 ml) followed by 15% sodium hydroxide (15 ml) and
water again (45 ml) was added slowly and the solution was filtered.
The oily residue was dissolved in diethyl ether and was purified by
acid-base partitioning (acetic acid-sodium hydroxide). The pure
amine was dissolved in ethanol and was hydrogenated over palladium
on carbon at atmospheric pressure. The catalyst was added in
portions because the amino-group deactivated the catalyst. When 1.2
ee hydrogen was consumed the hydrogenation was stopped. The
solution was filtered through Celite and was evaporated. The oily
product was dissolved in acetonitrile and 5-chloropyrid-2-yl
isothiocyanate (5 g) was added. The solution was refluxed for 20
minutes. The product separated on cooling and was collected on a
filter to yield 5.6 g of solid. The solid was dissolved in dioxane
and dilute hydrochloric acid was added. The solution was stirred
for 1 hour and was evaporated to dryness to yield 5.3 g of solid.
The solid was dissolved in dioxane containing 15% water. Silver
nitrate (25 g) dissolved in water was added. The solution was
stirred for 40 minutes, diluted with ethyl acetate, was washed with
water dried with sodium sulphate, filtered through Celite and
evaporated. The solid was recrystallized from acetonitrile to yield
the title product as a solid. .sup.1H-NMR DMSO-d.sub.6 .delta.2.6
(s, 3H) 2.9 (t, 2H) 3.4 (t, 2H) 6.8 (t, 1H) 7.4 (d, 1H) 7.7 (m, 1H)
7.9 (m, 1H) 9.2 (s, 1H) 11.3 (bs, 1H).
EXAMPLE 59
(+,-)-N-(cis-2-(6-chloro-3-ethoxy-2-fluorophenyl)-cyclopropyl)-N'-(5-chlor-
opyrid-2-yl)-urea
[0282] The starting material (+,
-)-cis-2-(6-chloro-3-ethoxy-2-fluoropheny- l) cyclopropylamine was
prepared from 4-chloro-2-fluorophenol in a manner analogous to
Examples 362, 375 and 348 of WO 93/03022.
5-Chloropyrid-2-yl-isothiocyanate was prepared as in Example 374 of
WO 93/03022 and then condensed with (+,
-)-cis-2-(6-chloro-3-ethoxy-2-fluoro- phenyl) cyclopropylamine in a
manner analogous to Example 370 of WO 93/03022 to give
(+,-)-N-(cis-2-(6-chloro-3-ethoxy-2-fluoro-phenyl)
cyclopropyl)-N'-(5-chloropyrid-2-yl)-thiourea. This compound was
reacted with NBS according to the procedure in Example 4 to provide
the titled compound. .sup.1H NMR (250 MHz, CDCl.sub.3)
.delta.1.29-1.38 (m, 1H), 1.45 (t, 1H), 1.57-1.67 (m, 1H), 2.09 (q,
1H), 3.27-3.37 (m, 1H), 4.05 (q, 2H), 6.78-6.85 (m, 2H), 7.14 (dd,
1H), 7.46 (dd, 1H), 7.81 (d, 1H), 9.27 (br s, 1H), 9.47 (br s,
1H).
EXAMPLE 60
(+,-)-N-(cis-2-(2,6-difluoro-3-ethoxyphenyl)
cyclopropyl)-N'-(5-chloropyri- d-2-yl)-urea
[0283] The starting material (+,
-)-cis-2-(2,6-difluoro-3-ethoxyphenyl)-cy- clopropylamine was
prepared from 2,4-difluorophenol in a manner analogous to Examples
362, 375 and 348 of WO 93/03022. 5-Chloropyrid-2-yl-isothiocy-
anate was prepared as in Example 374 of WO 93/03022 and then
condensed with (+,-)-cis-2-(2,6-difluoro-3-ethoxyphenyl)
cyclopropylamine in a manner analogous to Example 370 of WO
93/03022 to give
(+,-)-N-(cis-2-(2,6-difluoro-3-ethoxyphenyl)-cyclopropyl)
-N'-(5-chloropyrid-2-yl)-thiourea. This compound was reacted with
NBS according to the procedure in Example 4 to provide the titled
compound. .sup.1H NMR (250 MHz, CDCl.sub.3) .delta.1.31-1.38 (m,
1H), 1.45 (t, 3H), 1.49-1.58 (m, 1H), 2.06-2.15 (m, 1H), 3.26-3.35
(m, 1H), 4.04 (q, 2H), 6.72-6.85 (m, 3H), 7.48 (dd, 1H), 7.87 (d,
1H), 8.89 (br s, 1H), 9.24 (br s, 1H). Anal. Calc'd.. for
C.sub.17H.sub.16ClF.sub.2N.sub.3O.sub.2: C, 55.52; H, 4.38; N,
11.43. Found: C, 55.2; H, 4.3; N, 11.3.
EXAMPLE 61
(+,
-)-N-(cis-2-(3,6-dimethoxy-2-fluorophenyl)-cyclopropyl)-N'-(5-chloropy-
rid-2-yl)-urea
[0284] To a solution of 1,4-dimethoxybenzene (15.0 g, 0.109 mol) in
300 mL of dry THF was added 2.5 M n-butyllithium (45.6 mL, 0.114
mol) at room temperature under nitrogen.
[0285] After addition was complete, the solution was stirred for 1
hr. The mixture was cooled to -70.degree. C. and
N-fluorobenzenesulfonimide (36.0 g, 0.114 mol) in 150 mL of THF was
added slowly, keeping the temperature below -60.degree. C. The
solution was allowed to warm to room temperature during the night.
100 mL of NH.sub.4Cl (sat) was added and the mixture was extracted
with diethyl ether/THF. The organic phase was washed with 1 M NaOH
(2.times.60 mL), dried over MgSO.sub.4 and evaporated. Column
chromatography (silica gel, n-hexane followed by 1, 5 and 10% EtOAc
in n-hexane) provided 11.43 g of a mixture of
1,4-dimethoxy-2-fluorobenzene and 1,4-dimethoxybenzene (4.3:1).
This mixture was reacted in a manner analogous to Examples 362, 375
and 348 of WO 93/03022 to give (+,
-)-cis-2-(3,6-dimethoxy2-fluorophenyl) cyclopropylamine.
[0286] 5-Chloropyrid-2-ylisothiocyanate was prepared as in Example
374 of WO 93/03022 and then condensed with (+,
-)-cis-2-(3,6-dimethoxy-2-fluorop- henyl) cyclopropylamine in a
manner analogous to Example 370 of WO 93/03022 to give
(+,-)-N-(cis-2-(3,6-dimethoxy-2-fluorophenyl)-cyclopropy-
l)-N'-(5-chloropyrid-2-yl)-thiourea. This compound was reacted with
NBS according to the procedure in Example 4 to provide the titled
compound. .sup.1H NMR (250 MHz, CDCl.sub.3) .delta.1.23-1.30 (m,
1H), 1.46-1.56 (m, 1H), 1.97-2.07 (m, 1H), 3.21-3.32 (m, 1H), 3.80
(s, 3H), 3.83 (s, 3H), 6.59 (dd, 1H), 6.79-6.92 (m, 2H), 7.46 (dd,
1H), 7.75 (br s, 1H), 8.97 (br s, 1H), 9.20 (br s, 1H).
EXAMPLE 62
N-(2-(3,6-dimethoxy-2-fluorophenyl)ethyl-N'-(5-chloropyrid-2-yl)-urea
[0287] 1,4-dimethoxy-2-fluorobenzene was prepared as in Example 61
and was reacted in a manner analogous to Examples 362 and 151 of WO
93/03022 to give 3,6-dimethoxy-2-fluorophenethylamine.
5-Chloropyrid-2-ylisothiocyana- te was prepared as in Example 374
of WO 93/03022 and then condensed with
3,6-dimethoxy-2-fluorophenethylamine in a manner analogous to
Example 370 of WO 93/03022 to give
N-(2-(3,6-dimethoxy-2-fluorophenyl)ethyl)-N'-(5-ch-
loro-pyrid-2-yl)-thiourea. This compound was reacted with NBS
according to the procedure in Example 4 to provide the titled
compound. Anal. Calc'd.. for C.sub.16H.sub.17ClFN.sub.3O.sub.3: C,
54.32; H, 4.84; N, 11.88. Found: C, 53.8; H, 4.55; N, 11.65.
.sup.1H NMR (250 MHz, CDCl.sub.3) .delta.2.97-3.03 (m, 2H), 3.60
(q, 2H), 3.74 (s, 3H), 3.83 (s, 3H), 6.53 (dd, 1H), 6.79 (t, 1H),
6.88 (d, 1H), 7.52 (dd, 1H), 8.02 (d, 1H), 9.07 (br s, 1H), 9.51
(br s, 1H).
EXAMPLE 63
(+,
-)-N-(cis-2-(2-chloro-3-ethoxy-6-flourophenyl)-cyclopropyl)-N'-methyl--
N'-(5-chloropyrid-2-yl)-urea
[0288] 5-Chloro-2-methylaminopyridine was prepared as described by
Katritzky et al. in J. Chem. Soc. Perkin. Trans. I. 1987, 799-809.
The titled compound was obtained starting from
2-chloro-4-fluorophenol according to the procedure in Example 39.
The last step was made using
cis-2-(2-chloro-3-ethoxy-6-flourophenyl)cyclopropyl-carboxylic acid
and 5-Chloro-2-methylaminopyridine. .sup.1H NMR (250 MHz,
CDCl.sub.3) .delta.1.20-1.28 (m, 1H), 1.55 (t, 1H), 1.57-1.66 (m,
1H), 2.00-2.09 (m, 1H), 3.21-3.31 (m, 1H), 3.30 (s, 3H), 4.11 (q,
2H), 6.76-6.93 (m, 3H), 7.53 (dd, 1H), 7.86 (d, 1H), 10.0 (br s,
1H).
EXAMPLE 64
(+,
-)-N-(cis-2-(2-chloro-3-hydroxy-6-fluorophenyl)-cyclopropyl)-N'-(5-chl-
oropyrid-2-yl)-urea
[0289] The titled compound was obtained starting from
(+,-)-N-(cis-2-(2-chloro-3-ethoxy-6-fluorophenyl)-cyclopropyl)-N'-(5-chlo-
ropyrid-2-yl)-urea (Example 6) and BBr.sub.3 using the procedure in
Org. Synth., Coll Vol V, 1973, 412..sup.1H NMR (250 MHz,
DMSO-D.sub.6) .delta.1.10-1.18 (m, 1H), 1.54-1.62 (m, 1H),
2.10-2.20 (m, 1H), 3.20-3.30 (m, 1H), 6.99-7.16 (m, 2H), 7.32 (d,
1H), 7.84 (dd, 1H), 8.00 (d, 1H), 8.15 (br.s, 1H), 9.56 (br.s,
1H).
EXAMPLE 65
(+,
-)-N-(cis-2-(2-hydroxy-3-ethoxy-6-fluorophenyl)-cyclopropyl)-N'-(5-chl-
oropyrid-2-yl)-urea
[0290] 5'-Fluoro-2'-hydroxyacetophenone (9.38 g, 61 mmole),
iodoethane (7.3 ml, 91 mmole) and K.sub.2CO.sub.3 (12.6 g, 91
mmole) in acetone (100 ml) was stirred at 60.degree. C. overnight.
The mixture was filtered, evaporated, dissolved in hexane/ethyl
acetate, washed with 2M NaOH (2.times.20 ml) and water, dried
(MgSO.sub.4) and evaporated to give
2'-ethoxy-5'-fluoroacetophenone. A solution of mCPBA )19.3 g, 112
mmole) in CH.sub.2Cl.sub.2 (500 ml) was dried with MgSO.sub.4.
2'-Ethoxy-5'-fluoro-acetophenone (10.2 g, 56 mmole) was added and
the mixture was stirred for 3 days. More mCPBA (5 g, 29 mmole) and
MgSO.sub.4 was added and stirring was continued for 2 days. The
mixture was filtered, diluted with ethyl acetate, washed with 2 M
NaOH (2.times.70 ml), NH.sub.4Cl (sat, 50 ml), dried (MgSO.sub.4)
and evaporated. The residue (10.45 g,) was dissolved in ethanol
(100 ml). KOH (8.9 g, 160 mmole) was dissolved in water (50 ml).
The solutions were combined and stirred for lh. The mixture was
washed with diethyl ether/hexane 2.times.(50+25 ml), acidified with
HCl (konc), extracted with ethyl acetate (3.times.100 ml), dried
(MgSO.sub.4) and evaporated. The residue (8.22 g) was dissolved in
CH.sub.2Cl.sub.2 (100 ml). Triethylamine 11.7 ml, 84 mmole) and
bromomethyl methyl ether (6.45 ml, 79 mmole) were added and the
solution was refluxed for 2 hrs. More triethylamine 11.7 ml, 84
mmole) and bromomethyl methyl ether (6.45 ml, 79 mmole) were added
and refluxing was continued overnight. The solution was evaporated,
CH.sub.2Cl.sub.2 (200 ml) was added and the solution was washed
with water, 2M NaOH and NH.sub.4Cl. Drying (MgSO.sub.4) and
evaporation provided 4.0 g of
1-ethoxy-4-fluoro-2-methoxymethoxy-benzene. This compound was
reacted in a manner analogous to Example 39 to give
(+,-)-N-(cis-2-(3-ethoxy-6-fluoro-2-methoxymethoxyphenyl)-cyclopropyl)-N'-
-(5-chloropyrid-2-yl)-urea. Finally deprotection with 2 M HCl in
dioxane gave the titled compound. .sup.1H NMR (250 MHz,
DMSO-D.sub.6) .delta.1.04-1.14 (m, 1H), 1.41-1.56 (m+t, 4H),
1.96-2.06 (m, 1H), 3.05-3.15 (m, 1H), 4.06-4.19 (m, 2H), 6.64 (t,
1H), 6.94 (dd, 1H), 7.19 (d, 1H), 7.82(dd, 1H), 8.10(d, 1H), 8.61
(br.s, 1H), 9.08 (br.s, 1H), 9.57 (br.s, 1H).
EXAMPLE 66
(+, -)-N-[cis-2-(2-Chloro-3-ethoxy-6-fluorophenyl)
cyclopropyl]-N'-[2-(5-e- thynyl)pyridyl]-urea
[0291] Preparation of the intermediate, 2-Amino-5-ethynylpyridine.
2-Amin6-5-bromopyridine (8.65 g, 50.0 mmole) and
bis-trimethyl-silylaceta- mide (36.76 mL, 150.0 mmole) was stirred
at 100.degree. C. overnight. The mixture was purified by
distillation to give 10.41 g (85%) of the protected amine.
2-Trimethylsilylamino-5-bromopyridine (4.90 g, 20.0 mmole),
bis(triphenylphosphine)palladium-(II) chloride (0.70 g, 1.00 mmole)
and trimethylsilylacetylene (5.65 mL, 40.0 mmole) in piperidine (2
mL) was stirred under nitrogen at 80.degree. C. overnight. Diethyl
ether and NH.sub.4Cl (aq) was added to the mixture and the organic
phase was separated, dried over Na.sub.2SO.sub.4 and evaporated.
Tetrabutylammonium chloride (50.0 mL, 100.0 mmole) and THF (250 mL)
was added to remove the protection groups and the mixture was
stirred overnight.
[0292] THF was evaporated. The residue was purified by silica gel
column chromatography (EtOAc/Hexane, 40:60) to give 2.36 g, (61%)
of the titled compound. .sup.1H NMR (250 MHz, CDCl.sub.3)
.delta.3.05 (s, 1H), 4.65 (br s, 2H), 6.45 (d, 1H), 7.50 (dd, 1H),
8.25 (d, 1H).
[0293] (+,
-)-cis-(2-Chloro-3-ethoxy-6-fluorophenyl)-cyclopropylcarboxylic
acid (0,39 g, 1.5 mmole) which was made from
2-chloro-4-fluorophenol according to the procedure in Example 39,
was refluxed in toluene with diphenylphosphoryl azide (0.36 mL, 1.7
mmole) and triethylamine (0.23, 1.7 mmole) for 40 min.
2-Amino-5-ethynylpyridine (0.20 g, 1.7 mmole) was added in DMF (10
mL)and reflux was continued for 3 hrs. Toluene was evaporated. The
residue was dissolved in EtOAc and washed with HCl (0.1 M) and
water. The organic phase was dried over Na.sub.2SO.sub.4 and
evaporated. The residue was purified by silica gel column
chromatography (EtOAc/Hexane, 40:60-60:40) to give 0.16 (28%) of
the titled compound. Mp 197-198.degree. C. .sup.1H NMR (250 MHz,
CDCl.sup.3) .delta.1.2-1.8 (m, 5H), 2.1 (m, 1H), 3.1 (s, 1H), 3.3
(m, 1H), 4.1 (m, 2H), 6.7-7.0 (m, 3H), 7.6 (dd, 1H), 8.0 (d, 1H),
9.2 (s, 1H), 9.5 (br s, 1H).
EXAMPLE 67
(+, -)-N-[cis-2-(2-Chloro-3-ethoxy-6-fluorophenyl)
cyclopropyl]-N'-[2-(5-c- arboxy)pyridyl]-urea
[0294] (+,
-)-cis-(2-Chloro-3-ethoxy-6-fluorophenyl)-cyclopropylcarboxylic
acid (See Example 66), (0,26 g, 1.0 mmole) which was made from
2-chloro-4-fluorophenol according to the procedure in Example 39,
was refluxed in toluene with diphenylphosphoryl azide (0.24 mL, 1.1
mmole) and triethylamine (0.15, 1.1 mmole) for 40 min.
6-Aminonicotinic acid (0.15 g, 1.1 mmole) in DMF (10 mL) was added
and reflux was continued for 3 h. Toluene was evaporated. NaOH(aq)
was added and the mixture was washed with EtOAc. It was then made
acidic with conc HCl and extracted with EtOAc. The organic phase
was washed with water, dried over Na.sub.2SO.sub.4 and evaporated
to give 0.055 g (14%) of the titled compound. Mp 237-238.degree. C.
.sup.1H NMR (250 MHz, DMSO) .delta.1.20 (m, 1H), 1.50 (m, 3H), 1.60
(m, 1H), 2.20 (m, 1H), 3.25 (m, 1H), 3.40 (s, 1H), 4.20 (q, 2H),
7.20 (m, 3H), 8.15 (dd, 1H), 8.40 (s, 1H), 8.80 (br s, 1H), 9.85
(s, 1H).
EXAMPLE 68
(+, -)-N-[cis-2-(2-Chloro-3-ethoxy-6-fluorophenyl)
cyclopropyl]-N'-[2-(5-v- inyl)pyridyl]-urea
[0295] Preparation of the intermediate, 2-Amino-5-vinylpyridine.
Vinyltributyltin (3.1 mL, 10.5 mmole) was added dropwise to a
mixture of 2-trimethylsilyl-amino-5-bromopyridine (2.45 g, 10.0
mmole) from Example 66, LiCl (1.27 g, 30.0 mmole) and
tris-(dibenzylidene-acetone)dipalladium (0.18 g, 0.2 mmole) in DMF
(20 mL). The mixture was stirred overnight under nitrogen at
80.degree. C.
[0296] KF(aq) and EtOAc was added and the organic phase was
separated, dried over Na.sub.2SO.sub.4 and evaporated. The residue
was purified by silica gel chromatography (EtOAc/Hexane, 40: 60) to
give 0.12 g (29%) of the titled compound. .sup.1H NMR (250 MHz,
CDCl.sub.3) .delta.4.70 (br s, 2H), 5.10 (d, 1H), 5.55 (d, 1H),
6.40-6.65 (m, 2H), 7.55 (dd, 1H), 8.05 (s, 1H).
[0297] (+,
-)-cis-(2-Chloro-3-ethoxy-6-fluorophenyl)-cyclopropylcarboxylic
acid (0.39 g, 1.5 mmole) which was made from
2-chloro-4-fluorophenol according to the procedure in Example 39,
was refluxed in toluene with diphenylphosphoryl azide (0.36 mL, 1.7
mmole) and triethylamine (0.23 mL, 1.7 mmole) for 40 min.
2-Amino-5-vinylpyridine (0.20 g, 1.7 mmole) in DMF (10 mL) was
added and reflux was continued for 3 h. Toluene was evaporated. The
residue was dissolved in EtOAc and the solution was washed with HCl
(0.1M) and water, dried over Na.sub.2SO.sub.4 and evaporated. The
residue was purified by silica gel column chromatography
(EtOAc/Hexane, 40: 60-60:40) and recrystallized from acetonitrile
to give 0.12 g (21%) of the titled compound. .sup.1H NMR (250 MHz,
CDCl.sub.3) .delta.1.40 (m, 1H), 1.50 (t, 3H), 1.65 (m, 1H), 2.10
(m, 1H), 3.45 (m, 1H), 4.10 (q, 2H), 5.25 (d, 1H), 5.60 (d, 1H),
6.50-6.95 (m, 4H), 7.60 (dd, 1H), 7.70 (s, 1H), 8.90 (s, 1H), 9.50
(br s, 1H).
EXAMPLE 69
(+, -)-N-[cis-2-(2-Chloro-3-ethoxy-6-fluorophenyl)
cyclopropyl]-N'-[2-(5-b- romo)pyrazyl]-urea
[0298] Preparation of the intermediate 2-Amino-5-bromopyrazine.
Br.sub.2 (0.61 mL, 12.0 mmole) was added dropwise to
2-aminopyrazine (0.95 g, 10.0 mmole) in pyridine (10 mL). The
mixture was stirred at 30.degree. C. for 1 h and at 70.degree. C.
for 45 min. CH.sub.2Cl.sub.2 was added and the mixture was washed
with water and NaOH (1M), filtered, dried over Na.sub.2SO.sub.4 and
evaporated. The residue was purified by silica gel column
chromatography (EtOAc/Hexane, 30:70-40:60) to give 0.50 g (29%)of
the titled compound. .sup.1H NMR (250 MHz, DMSO) .delta.6.80 (s,
2H), 7.80 (s, 1H), 8.15 (s, 1H).
[0299] (+,
-)-cis-(2-Chloro-3-ethoxy-6-fluorophenyl)-cyclopropylcarboxylic
acid (0.52 g, 2.0 mmole) which was made from
2-chloro-4-fluorophenol according to the procedure in Example 39,
was refluxed in toluene with diphenylphosphoryl azide (0.47 mL, 2.2
mmole) and triethylamine (0.3OmL, 2.2 mmole) for 30 min.
2-Amino-5-bromopyrazine (0.38 g, 2.2 mmole) was added in DMF (10
mL) and reflux was continued overnight. The mixture was then
stirred without heat for another 24 hrs.
[0300] Toluene was evaporated. The residue was dissolved in EtOAc
and the solution was washed with HCl (0.1M) and water, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was filtered using
EtOAc as eluent. The filtrate was evaporated and the residue was
further purified by recrystallization in acetonitrile to give 0.088
g (10%) of the titled compound. .sup.1H NMR (250 MHz, CDCl.sub.3)
.delta.1.30 (m, 1H), 1.55 (m, 3H), 1.65 (m, 1H), 1.30 (m, 1H), 3.25
(m, 1H), 4.10 (q, 2H), 6.75-6.95, (m, 2H), 7.90 (s, 1H), 8.10 (s,
1H), 9.25 (s, 1H).
EXAMPLE I
N-(2-(2.6-Difluorophenethyl)-N'-(5-bromopyrid-2-yl)-quanidinium
hydrochloride
[0301] To a solution of 5.0 g (32.65 mmole) of
2,6-difluorophenylacetonitr- ile in 25 ml of anhydrous
tetrahydrofuran at room temperature under nitrogen was added 33.0
ml (33.0 mmole) of 1.0 molar borane-tetrahydrofuran complex by
syringe. The mixture was stirred 4 hours, cooled in an ice bath,
and quenched under nitrogen with a solution of 10.7 ml of
concentrated hydrochloric acid in 22 ml of water. The mixture was
heated to reflux, stirred I hr., and the tetrahydrofuran was
removed by downward distillation. The milky white suspension was
diluted with 22 ml of toluene and stirred 5 minutes longer. The
layers were separated and the aqueous was extracted with 20 ml of
hot toluene. The aqueous layer was cooled to 0.degree. C., 50 ml of
methylene chloride and 10 ml of 50% sodium hydroxide was added, and
the layers were separated. The aqueous layer was extracted with
methylene chloride (2.times.) and the combined organics were dried
over sodium sulfate. Concentration gave 2.8 g (55%) of
2,6difluorophenethylamine as a light yellow liquid; .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta.7.10-7.30 (m, 1H), 6.80-7.00 (t, 2H),
2.80-3.00 (m, 2H), 2.70-2.80 (m, 2H), 1.35 (s, 2H).
[0302] The amine prepared as above (2,6 g, 16.56 mmole) was
dissolved in 50 ml of anhydrous ethyl ether and cooled with
stirring in an ice water bath. 1.1 g (10.3 mmole) of cyanogen
bromide dissolved in 15 ml of ethyl ether was added by dropping
funnel and the mixture was stirred 1 hr. The resulting solid (2,6
difluorophenethylamine hydrobromide) was filtered off.
Concentration of the filtrate gave 1.5 g (50%) of
2,6-difluorophenethyl cyanoacetamide as a yellow off: .sup.1H NMR
(300 MHz, DMSO-d.sub.6) .delta.7.20-7.40 (m, 1H), (6.80-7.00 (m,
2H), 4.00 (s, 1H), 3.26 (q, 2H), 3.00 (t, 2H).
[0303] 5.0 g (28.8 mmole) of 2-amino-5-bromopyridine was dissolved
in 50 ml of ethanol and cooled to 0.degree. C. with stirring.
Hydrogen chloride (g) was bubbled in until the solution was
saturated, the ethanol was concentrated to a small volume, ethyl
ether was added, and the resulting solid was filtered and dried to
give 4.0 g (66%) of 2-amino-5-bromopyridine hydrochloride: .sup.1H
NMR (300 MHz, DMSO-d.sub.6 .delta.8.20 (s, 1H), 8.00 (d, 1H), 7.00
(d, 1H), 3.50-5.50 (br, 3H).
[0304] 750 mg (4.12 mmole) of 2,6-difluorophenethyl cyanoacetamide
and 865 mg (4.12 mmole) of 2-amino-5-bromopyridine hydrochloride
prepared as above in 20 ml of chlorobenzene was heated at
125.degree. C. under nitrogen for 3.5 hrs. The solution was cooled
to room temperature and the resulting solid was filtered.
Recrystallization from ethanol/ethyl ether gave 540 mg (33.5%) of
the titled compound as a white solid: mp 209-211.degree. C.; IR
(KBr, cm.sup.-1) 3094, 1680, 1627, 1468, 1236, 1000, 829, 775;
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.11.45 (br.s., 1H), 9.30
(br.s., 1H), 8.60 (br.s., 2H), 8.30 (d, 1H), 8.10 (dd, 1H),
7.30-7.45 (m, 1H), 7.00-7.20 (m, 3H), 3.60 (q, 2H), 2.95 (t, 2H);
MS (FD) m/e 356 (Mt, free base). Anal. Calc'd.. for
C.sub.14H.sub.13BrF.sub.2N.su- b.4 HCl 0.25 H.sub.20: C, 42.45; H,
3.66; N, 14.14.
[0305] Found: C, 42.53; H, 4.04; N, 14.02.
EXAMPLE II
N-(3-Fluoropyrid-2-yl)eth-2yl-N'-(5-bromopyrid-2-yl)-quanidine
[0306] A mixture of 500 mg (3.03 mmole) of N-(3-fluoropyrid-2-yl)
eth-2-yl-cyanamide (prepared from
N-2-(3-fluoropyrid-2-yl)ethylamine as in Example I), 635 mg (3.03
mmole) of 2-amino-5-bromopyridine hydrochloride (prepared as in
Example I), and 15 ml of chlorobenzene was heated at 125.degree. C.
under nitrogen for 4 hrs. The mixture was cooled to room
temperature, stirred overnight, and concentrated to dryness. The
resin was dissolved in 40 ml of IN hydrochloric acid and extracted
with ethyl acetate (IX). The aqueous layer was basified with 2N
sodium hydroxide and extracted with ethyl acetate (2.times.). The
organic layer was dried over sodium sulfate, concentrated to a
resin, and purified by flash silica gel chromatography (methylene
chloride/methanol/ammonium hydroxide -46:3:1) to give a yellow
oil.
[0307] Crystallization from ethyl ether gave the titled compound as
a pale yellow solid, 200 mg (19.5%): mp 134-137.degree. C.; IR
(KBr, cm.sup.-1) 3400, 3070, 1679, 1573, 1450, 1127, 827, 710;
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.8.40 (d, 1H), 8.07 (d,
1H), 7.70-7.90 (m, 2H), 7.30-7.50 (m, 3H), 6.80-7.00 (br, 1H), 6.60
(d, 1H), 3.60 (br.q., 2H), 3.00 (dt, 2H); MS (FD) m/e 337 (Mt).
Anal. Calc'd.. for C.sub.13H.sub.13BrFN.sub.5: C, 46.17; H, 3.88;
N, 20.71. Found: C, 46.13; H, 3.68; N, 20.92.
EXAMPLE III
N-(2-(2-Chloro-3-ethoxy-6-fluorophenethyl)-N'-(5-bromopyrid-2-yl)-guanidin-
e
[0308] A solution of 380 mg (1.81 mmole) of 2-amino-5-bromopyridine
hydrochloride and 440 mg (1.8 mmole) of
N-(2-(2-chloro-3-ethoxy-6-fluorop- henethyl) cyanamide (prepared as
in Example 1) in 15 ml of chlorobenzene was heated at 125.degree.
C. for 4 hrs. under nitrogen. The mixture was cooled to room
temperature, stirred overnight, and concentrated to dryness. The
residue was partitioned between 40 ml of IN hydrochloric acid and
ethyl acetate (IX). The organic layer was extracted with 2N sodium
hydroxide (IX), dried over sodium sulfate, and concentrated to an
orange oil. Purification over a flash silica gel column (methylene
chloride/methanol/ammonium hydroxide -92:7:1) provided a yellow
oil.
[0309] Crystallization from ethyl ether gave the titled compound as
an off-white solid, 100 mg (13%): mp 169-172.degree. C.; IR (KBr,
cm.sup.-1) 3400, 3100, 1668, 1568, 1465, 1243, 1072, 826, 805;
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.8.10 (d, 1H), 7.60 (dd,
1H), 7.35 (br, 2H), 7.10-7.20 (m, 1H), 7.00-7.05 (m, 1H), 6.70-6.90
(br, 1H), 6.60 (d, 1H), 4.05 (q, 2H), 3.40 (m, 2H), 3.00 (t, 2H),
1.35 (t, 3H); MS (FD) m/e 416 (Mt). Anal. Calc'd.. for
C.sub.16H.sub.17BrClFN.sub.40: C, 46.23; H, 4.12; N, 13.48. Found:
C, 46.41; H, 4.18; N, 13.32.
EXAMPLE IV
N-(2-Phenethyl) -N'-(5-bromopyrid-2-yl) -guanidinium
hydrochloride
[0310] The titled compound was prepared from
N-(2-phenethyl)cyanamide and 2-amino-5-bromopyridine hydrochloride
as in Example I: mp 176-178.degree. C.; IR (KBr, cm.sup.-1) 3252,
3100, 1677, 1628, 1586, 1468, 1365, 1235, 826, 705; .sup.1H NMR
(300 MHz, DMSO-d.sub.6) .delta.11.5 (br, 1H), 9.30 (br, 1H), 8.55
(br, 2H), 8.35 (d, 1H), 8.06 (dd, 1H), 7.20-7.40 (m, 5H), 7.05
(br.d., 1H), 3.60 (q, 2H), 2.90 (t, 2H); MS (FD) m/e 320 (Mt).
Anal. Calc'd.. for C.sub.14H.sub.15BrN.sub.4 HCl: C, 47.28; H,
4.53; N, 15.75. Found: C, 47.48; H, 4.63; N, 15.81.
EXAMPLE V
N-(2-(2-Fluorophenethyl)-N'-(5-bromopyrid-2-yl)-quanidirlium
hydrochloride
[0311] The titled compound was prepared from
N-(2-(2-fluorophenethyl) cyanamide and 2-amino-5-bromopyridine
hydrochloride according to Example I: mp 175-177.degree. C.; IR
(KBr, cm.sup.-1) 3090, 2933, 1679, 1622, 1471, 1227, 833, 759, 635;
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.11.48 (br, 1H), 9.30
(br, 1H), 8.55 (br.s., 2H), 8.35 (d, 1H), 8.10 (dd, 1H), 7.40-7.50
(dt, 1H), 7.25-7.38 (m, 1H), 7.20 (t, 2H), 7.06 (br.d., 1H), 3.62
(q, 2H), 2.86 (t, 2H); MS (FD) m/e 336 (Mt). Anal. Calc'd. for
C.sub.14H.sub.14BrFN.sub.4 HCl: C, 45.00; H, 4.05; N, 14.99. Found:
C, 45.20; H, 4.03; N, 15.04.
EXAMPLE VI
N-(2-(2-Pyridylethyl)-N'-(5-bromopyrid-2-yl) quanidine
[0312] The titled compound was prepared from N-(2-(2-pyridylethyl)
cyanamide and 2-amino-5-bromopyridine hydrochloride as in Example
II: mp 139-141.degree. C.; IR (KBr, cm.sup.-1) 3600, 3082, 1681,
1566.
EXAMPLE VII
N-Cyano-{N'-2-phenethyl-N"-[2-(5-chloro)pyridyl]}quanidine
[0313] A mixture of Na.sub.2NCN (95%, 0.25 g, 2.78 mmole) and
Et.sub.3N HCl (0.38 g, 2.78 mmole) in EtOH (5 mL) was stirred at
room temperature overnight. 2-(5-chloro)pyridyl isothiocyanate
(0.48 g, 2.78 mmole), the product of Example 374 of WO 93/03022,
was added to the reaction mixture and it was stirred for 1 hr. at
room temperature.
[0314] The solvent was evaporated and 5 mL of DMF and
phenethylamine (0.42 mL, mL, 3.34 mmole) added to the residue. The
mixture was stirred for 3 min. at room temperature,
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide HCl (0.64 g, 3.34
mmole) was added to it and stirring was continued for 0.5 hr. at
room temperature. 30 mL of EtOAc was added to the mixture and it
was washed once with 10 mL of 1N HCl and three times with 10 mL of
H.sub.20. The collected water phase was made alkaline with 15% NaOH
(aq) and extracted with EtOAc which was dried over Na.sub.2SO.sub.4
and evaporated to yield the titled product. Mp 189-190.degree. C.
.sup.1H NMR (250 MHz, CDCl.sub.3) .delta.10.08 (s, 1H), 9.04 (s,
1H), 7.93 (d, 1H), 7.60 (dd, 1H), 7.34-7.21 (m, 6H), 3.69 (q, 2H),
2.92 (t, 2H).
EXAMPLE VIII
(+,-)-N-(cis-2-phenylcyclopropyl)-N'-[2-(5-bromo)pyridyl]quanidine
[0315]
(+,-)-N-(cis-2-phenylcyclopropyl)-N'-[2-(5-bromo)pyridyl]-thiourea
(0.2 g, 0.57 mmole), the product of Example 373 of WO 93/03022, and
silver triflate (0.22 g, 0.86 mmole) were dissolved in 20 mL of
CH.sub.2Cl.sub.2 at -20.degree. C.
[0316] NH.sub.3 (g) was introduced to this solution at the same
temperature. The mixture was stirred and the temperature was
allowed to rise slowly to room temperature. Stirring was continued
overnight. The mixture was filtrated and the filtrate was
evaporated. The residue was stirred with 4N HCl and filtrated. The
filtrate was made alkaline and extracted with CH.sub.2Cl.sub.2
which was dried over Na.sub.2SO.sub.4 and evaporated to give the
titled product. .sup.1H NMR (250 MHz, CDCl.sub.3) .delta.7.82 (br
s, 1H), 7.49 (dd, 1H), 7.35-7.22 (m, 6H), 6.66 (d, 1H), 2.91 (m,
1H), 2.40 (q, 1H), 1.42 (m, 1H), 1.18 (m, 1H).
* * * * *