U.S. patent number RE44,613 [Application Number 13/281,965] was granted by the patent office on 2013-11-26 for n-substituted glycine derivatives: hydroxylase inhibitors.
This patent grant is currently assigned to GlaxoSmithKline LLC. The grantee listed for this patent is Kevin J. Duffy, William Henry Miller, Andrea K. Myers, Antony N. Shaw, Michael N. Zimmerman. Invention is credited to Kevin J. Duffy, William Henry Miller, Andrea K. Myers, Antony N. Shaw, Michael N. Zimmerman.
United States Patent |
RE44,613 |
Duffy , et al. |
November 26, 2013 |
N-substituted glycine derivatives: hydroxylase inhibitors
Abstract
The invention described herein relates to certain
pyridazinedione N-substituted glycine derivatives of formula (I)
##STR00001## which are antagonists of HIF prolyl hydroxylases and
are useful for treating diseases benefiting from the inhibition of
this enzyme, anemia being one example.
Inventors: |
Duffy; Kevin J. (Collegeville,
PA), Miller; William Henry (Collegeville, PA), Myers;
Andrea K. (Collegeville, PA), Shaw; Antony N. (King of
Prussia, PA), Zimmerman; Michael N. (Collegeville, PA) |
Applicant: |
Name |
City |
State |
Country |
Type |
Duffy; Kevin J.
Miller; William Henry
Myers; Andrea K.
Shaw; Antony N.
Zimmerman; Michael N. |
Collegeville
Collegeville
Collegeville
King of Prussia
Collegeville |
PA
PA
PA
PA
PA |
US
US
US
US
US |
|
|
Assignee: |
GlaxoSmithKline LLC
(Philadelphia, PA)
|
Family
ID: |
39636631 |
Appl.
No.: |
13/281,965 |
Filed: |
October 26, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
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60884710 |
Jan 12, 2007 |
|
|
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Reissue of: |
11972702 |
Jan 11, 2008 |
7608621 |
Oct 27, 2009 |
|
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Current U.S.
Class: |
514/247;
544/240 |
Current CPC
Class: |
A61P
7/00 (20180101); A61P 7/06 (20180101); C07D
237/24 (20130101); A61P 9/10 (20180101); A61P
43/00 (20180101) |
Current International
Class: |
A01N
43/58 (20060101); A61K 31/50 (20060101) |
Field of
Search: |
;544/240 ;514/247 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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03/088917 |
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Oct 2003 |
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WO |
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2006/048330 |
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May 2006 |
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WO |
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2008/013838 |
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Jan 2008 |
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WO |
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2008/040002 |
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Apr 2008 |
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WO |
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2009/142732 |
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Nov 2009 |
|
WO |
|
Other References
Nemeth, Advances in Hematology, 2012, Article ID 750643, 9 pages.
cited by examiner .
Haase, Am. J. Physiol. Renal Physiol. 291: F271-F281, 2006. cited
by examiner .
Kaelin, Biochem. & Biophys. Research Commun., 338 (2005)
627-638. cited by examiner .
M.A. Goldberg et al., 84 PNAS, 7972-7976 (1987). cited by examiner
.
Y. Setoguchi et al., 84 Blood, 2946-2953 (1994). cited by examiner
.
V. Bardet et al., 12 Liver Transplantation 1649-1654 (2006). cited
by examiner .
I.C. Macdougall, Seminars in Nephrology, 313-318 (2006). cited by
examiner .
Auerbach et al, 22 Journal of Clinical Oncology 1301-1307 (2004).
cited by examiner .
Mikhail et al., "Stimulating Erythropoiesis: Future Perspectives,"
Kidney Blood Press. Res., 2008, 31(4), pp. 234-246. cited by
applicant .
Mikhail, et al., Kidney & Blood Pressure Research,
2008;31:234-246. cited by examiner.
|
Primary Examiner: Wilson; James O
Assistant Examiner: Pagano; Alexander R
Attorney, Agent or Firm: Coulter; Kathryn L. Peng; Tony
W.
Parent Case Text
.Iadd.CROSS REFERENCE TO RELATED APPLICATIONS .Iaddend.
.Iadd.This application is a reissue application of U.S. application
Ser. No. 11/972,702, filed Jan. 11, 2008, now U.S. Pat. No.
7,608,621, which claims the benefit of U.S. Provisional Application
No. 60/884,710, filed Jan. 12, 2007. .Iaddend.
Claims
What is claimed is:
1. A compound which is
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1
-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine or a
salt thereof.
2. A compound according to claim 1 which is
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine.
3. A compound according to claim 1 which is Na, K, Li, Ca, or Mg
salt of
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine.
.[.4. A method for treating anemia in a mammal, which method
comprises administering an effective amount of
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine or a salt thereof to a
mammal suffering from anemia which can be treated by inhibiting HIF
prolyl hydroxylases..].
5. A pharmaceutical composition comprising
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine or a salt thereof and
one or more of pharmaceutically acceptable carriers, diluents and
excipients.
6. A compound which is
N-{[2-[(3,4'-difluoro-4-biphenylyl)methyl]-5-hydroxy-6-(1-methylethyl)-3--
oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine or a
pharmaceutically acceptable salt thereof.
7. A compound according to claim 6 which is
N-{[2-[(3,4'-difluoro-4-biphenylyl)methyl]-5-hydroxy-6-(1
-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine.
8. A compound according to claim 6 which is the Na, K, Li, Ca, or
Mg salt of
N-{[2-[(3,4'-difluoro-4-biphenylyl)methyl]-5-hydroxy-6-(1-methylethyl)-
-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine.
.[.9. A method for treating anemia in a mammal, which method
comprises administering an effective amount of
N-{[2-[(3,4'-difluoro-4-biphenylyl)methyl]-5-hydroxy-6-(1-methylethyl)-3--
oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine or a salt thereof to
a mammal suffering from anemia which can be treated by inhibiting
HIF prolyl hydroxylases..].
10. A pharmaceutical composition comprising
N-{[2-[(3,4'-difluoro-4-biphenylyl)methyl]-5-hydroxy-6-(1
-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine or a
salt thereof and one or more of pharmaceutically acceptable
carriers, diluents and excipients.
.Iadd.11. A compound of Formula (I) ##STR00121## wherein R.sup.1 is
2-fluoro-4-bromobenzyl, 4-bromobenzyl, 4-(halophenyl)benzyl,
2-fluoro-4-(halophenyl)benzyl, 2-fluoro-4-trifluoromethylbenzyl,
4-trifluoromethylbenzyl, or
2-fluoro-4-(C.sub.1-C.sub.4alkoxyphenyl)benzyl; R.sup.2 is OH;
R.sup.3 is H; and R.sup.4 is isopropyl, t-butyl or cyclohexyl; or a
pharmaceutically acceptable salt thereof..Iaddend.
.Iadd.12. A pharmaceutical composition comprising a compound of
claim 11, or a salt thereof and one or more of pharmaceutically
acceptable carriers, diluents or excipients..Iaddend.
.Iadd.13. The compound: ##STR00122## or a salt
thereof..Iaddend.
.Iadd.14. The compound: ##STR00123## .Iaddend.
.Iadd.15. A pharmaceutical composition comprising a compound
according to claim 13, and one or more of pharmaceutically
acceptable carriers, diluents or excipients..Iaddend.
.Iadd.16. The compound: ##STR00124## or a salt
thereof..Iaddend.
.Iadd.17. The compound: ##STR00125## .Iaddend.
.Iadd.18. A pharmaceutical composition comprising a compound of
claim 16, and one or more of pharmaceutically acceptable carriers,
diluents or excipients..Iaddend.
.Iadd.19. A method for treating anemia associated with renal
disease in a human, said method comprising administering a compound
according to claim 13..Iaddend.
.Iadd.20. A method for treating anemia associated with renal
disease in a human, said method comprising administering a compound
according to claim 16..Iaddend.
.Iadd.21. A method for treating anemia associated with cancer
chemotherapy in a human, said method comprising administering a
compound according to claim 13..Iaddend.
.Iadd.22. A method for treating anemia associated with cancer
chemotherapy in a human, said method comprising administering a
compound according to claim 16..Iaddend.
.Iadd.23. A method for treating anemia associated with reduced
erythropoietin production in a human, said method comprising
administering a compound according to claim 13..Iaddend.
.Iadd.24. A method for treating anemia associated with reduced
erythropoietin production in a human, said method comprising
administering a compound according to claim 16..Iaddend.
.Iadd.25. A method for increasing erythropoietin production in a
human, said method comprising administering a compound according to
claim 13..Iaddend.
.Iadd.26. A method for increasing erythropoietin production in a
human, said method comprising administering a compound according to
claim 16..Iaddend.
Description
FIELD OF THE INVENTION
This invention relates to certain heteroaromatic N-substituted
glycine derivatives that are inhibitors of HIF prolyl hydroxylases,
and thus have use in treating diseases benefiting from the
inhibition of this enzyme, anemia being one example.
BACKGROUND OF THE INVENTION
Anemia occurs when there is a decrease or abnormality in red blood
cells, which leads to reduced oxygen levels in the blood. Anemia
occurs often in cancer patients, particularly those receiving
chemotherapy. Anemia is often seen in the elderly population,
patients with renal disease, and in a wide variety of conditions
associated with chronic disease.
Frequently, the cause of anemia is reduced erythropoietin (Epo)
production resulting in prevention of erythropoiesis (maturation of
red blood cells). Epo production can be increased by inhibition of
prolyl hydroxylases that regulate hypoxia inducible factor
(HIF).
One strategy to increase erythropoietin (Epo) production is to
stabilize and thus increase the transcriptional activity of the
HIF. HIF-alpha subunits (HIF-1alpha, HIF-2alpha, and HIF-3alpha)
are rapidly degraded by proteosome under normoxic conditions upon
hydroxylation of proline residues by prolyl hydroxylases (EGLN1, 2,
3). Proline hydroxylation allows interaction with the von Hippel
Lindau (VHL) protein, a component of an E3 ubiquitin ligase. This
leads to ubiquitination of HIF-alpha and subsequent degradation.
Under hypoxic conditions, the inhibitory activity of the prolyl
hydroxylases is suppressed, HIF-alpha subunits are therefore
stabilized, and HIF-responsive genes, including Epo, are
transcribed. Thus, inhibition of prolyl hydroxylases results in
increased levels of HIF-alpha and thus increased Epo
production.
The compounds of this invention provide a means for inhibiting
these hydroxylases, increasing Epo production, and thereby treating
anemia. Ischemia, stroke, and cytoprotection may also benefit by
administering these compounds.
SUMMARY OF THE INVENTION
In the first instance, this invention relates to a compound of
formula (I):
##STR00002## wherein:
R.sup.1 is selected from the group consisting of hydrogen,
--NR.sup.5R.sup.6, C.sub.1-C.sub.10alkyl, C.sub.2-C.sub.10alkenyl,
C.sub.2-C.sub.10alkynyl, C.sub.3-C.sub.8cycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8cycloalkyl,
C.sub.5-C.sub.8cycloalkenyl, C.sub.1-C.sub.10alkyl-C.sub.5-C.sub.8
cycloalkenyl, C.sub.3-C.sub.8 heterocycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8 heterocycloalkyl, aryl,
C.sub.1-C.sub.10alkyl-aryl, heteroaryl and
C.sub.1-C.sub.10alkyl-heteroaryl;
R.sup.4 is selected from the group consisting of hydrogen,
COOR.sup.9, CONR.sup.7R.sup.8, --NR.sup.5R.sup.6,
C.sub.1-C.sub.10alkyl, C.sub.2-C.sub.10alkenyl,
C.sub.2-C.sub.10alkynyl, C.sub.3-C.sub.8cycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8cycloalkyl,
C.sub.5-C.sub.8cycloalkenyl, C.sub.1-C.sub.10alkyl-C.sub.5-C.sub.8
cycloalkenyl, C.sub.3-C.sub.8 heterocycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8 heterocycloalkyl, aryl,
C.sub.1-C.sub.10alkyl-aryl, heteroaryl and
C.sub.1-C.sub.10alkyl-heteroaryl;
R.sup.2 is NR.sup.7R.sup.8 or --OR.sup.9;
R.sup.3 is H or C.sub.1-C.sub.4alkyl;
R.sup.5 and R.sup.6 are each independently selected from the group
consisting of hydrogen, C.sub.1-C.sub.10 alkyl,
C.sub.3-C.sub.8cycloalkyl, C.sub.1-C.sub.10
alkyl-C.sub.3-C.sub.8cycloalkyl, C.sub.3-C.sub.8heterocycloalkyl,
C.sub.1-C.sub.10 alkyl-C.sub.3-C.sub.8heterocycloalkyl, aryl,
C.sub.1-C.sub.10alkyl-aryl, heteroaryl,
C.sub.1-C.sub.10alkyl-heteroaryl, --CO(C.sub.1-C.sub.4 alkyl),
--CO(C.sub.3-C.sub.6 cycloalkyl), --CO(C.sub.3-C.sub.6
heterocycloalkyl), --CO(aryl), --CO(heteroaryl), and
--SO.sub.2(C.sub.1-C.sub.4 alkyl); or R.sup.5 and R.sup.6 taken
together with the nitrogen to which they are attached form a 5- or
6- or 7-membered saturated ring optionally containing one other
heteroatom selected from the group consisting of oxygen, nitrogen
and sulphur;
R.sup.7 and R.sup.8 are each independently selected from the group
consisting of hydrogen, C.sub.1-C.sub.10 alkyl, C.sub.2-C.sub.10
alkenyl, C.sub.2-C.sub.10 alkynyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.3-C.sub.8 heterocycloalkyl, aryl and heteroaryl;
R.sup.9 is H or a cation, or C.sub.1-C.sub.10alkyl which is
unsubstituted or substituted with one or more substituents
independently selected from the group consisting of C.sub.3-C.sub.6
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl; and wherein any
carbon or heteroatom of R.sup.1, R.sup.2, R.sup.3, R.sup.4,
R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9 is unsubstituted or,
where possible, is substituted with one or more substituents
independently selected from the group consisting of C.sub.1-C.sub.6
alkyl, aryl, heteroaryl, halogen, --OR.sup.10, NR.sup.5R.sup.6,
cyano, nitro, --C(O)R.sup.10, --C(O)OR.sup.10, --SR.sup.10,
--S(O)R.sup.10, --S(O).sub.2R.sup.10, --NR.sup.5R.sup.6,
--CONR.sup.5R.sup.6, --N(R.sup.5)C(O) R.sup.10,
--N(R.sup.5)C(O)OR.sup.10, --OC(O)NR.sup.5R.sup.6, --N(R.sup.5)C
(O)NR.sup.5R.sup.6, --SO.sub.2NR.sup.5R.sup.6,
--N(R.sup.5)SO.sub.2R.sup.10, C.sub.1-C.sub.10alkenyl,
C.sub.1-C.sub.10alkynyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.3-C.sub.6 heterocycloalkyl, aryl and heteroaryl group;
wherein R.sup.5 and R.sup.6 are the same as defined above and
R.sup.10 is hydrogen, C.sub.1-C.sub.10alkyl,
C.sub.2-C.sub.10alkenyl, C.sub.2-C.sub.10alkynyl, --CO
(C.sub.1-C.sub.4 alkyl), --CO(aryl), --CO(heteroaryl), --CO
(C.sub.3-C.sub.6 cycloalkyl), --CO(C.sub.3-C.sub.6
heterocycloalkyl), --SO.sub.2(C.sub.1-C.sub.4 alkyl),
C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8heterocycloalkyl,
C.sub.6-C.sub.14 aryl, C.sub.1-C.sub.10alkyl-aryl, heteroaryl, or
C.sub.1-C.sub.10alkyl-heteroaryl; or a pharmaceutically acceptable
salt or solvate thereof.
In a second aspect of the present invention, there is provided a
compound of formula (I) or a salt or solvate thereof for use in
mammalian therapy, e.g. treating amenia. An example of this
therapeutic approach is that of a method for treating anemia caused
by increasing the production of erythropoietin (Epo) by inhibiting
HIF prolyl hydroxylases comprising administering a compound of
formula (I) to a patient in need thereof, neat or admixed with a
pharmaceutically acceptable excipient, in an amount sufficient to
increase production of Epo.
In a third aspect of the present invention, there is provided a
pharmaceutical composition comprising a compound of formula (I) or
a salt, solvate, or the like thereof, and one or more of
pharmaceutically acceptable carriers, diluents and excipients.
In a fourth aspect, there is provided the use of a compound of
formula (I) or a salt or solvate thereof in the preparation of a
medicament for use in the treatment of a disorder mediated by
inhibiting HIF prolyl hydroxylases, such as an anemia, that can be
treated by inhibiting HIF prolyl hydroxylases.
DETAILED DESCRIPTION OF THE INVENTION
For the avoidance of doubt, unless otherwise indicated, the term
"substituted" means substituted by one or more defined groups. In
the case where groups may be selected from a number of alternative
groups the selected groups may be the same or different.
The term "independently" means that where more than one substituent
is selected from a number of possible substituents, those
substituents may be the same or different.
An "effective amount" means that amount of a drug or pharmaceutical
agent that will elicit the biological or medical response of a
tissue, system, animal or human that is being sought, for instance,
by a researcher or clinician. Furthermore, the term
"therapeutically effective amount" means any amount which, as
compared to a corresponding subject who has not received such
amount, results in improved treatment, healing, prevention, or
amelioration of a disease, disorder, or side effect, or a decrease
in the rate of advancement of a disease or disorder. The term also
includes within its scope amounts effective to enhance normal
physiological function.
As used herein the term "alkyl" refers to a straight- or
branched-chain hydrocarbon radical having the specified number of
carbon atoms, so for example, as used herein, the terms
"C.sub.1-C.sub.4 alkyl" and "C.sub.1-C.sub.10 alkyl" refers to an
alkyl group having at least 1 and up to 4 or 10 carbon atoms
respectively. Examples of such branched or straight-chained alkyl
groups useful in the present invention include, but are not limited
to, methyl, ethyl, n-propyl, isopropyl, isobutyl, n-butyl, t-butyl,
n-pentyl, isopentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, and
n-decyl, and branched analogs of the latter 5 normal alkanes.
When the term "alkenyl" (or "alkenylene") is used it refers to
straight or branched hydrocarbon chains containing the specified
number of carbon atoms and at least 1 and up to 5 carbon-carbon
double bonds. Examples include ethenyl (or ethenylene) and propenyl
(or propenylene).
When the term "alkynyl" (or "alkynylene") is used it refers to
straight or branched hydrocarbon chains containing the specified
number of carbon atoms and at least 1 and up to 5 carbon-carbon
triple bonds. Examples include ethynyl (or ethenylene) and propynyl
(or propenylene).
When "cycloalkyl" is used it refers to a non-aromatic, saturated,
cyclic hydrocarbon ring containing the specified number of carbon
atoms. So, for example, the term "C.sub.3-C.sub.8 cycloalkyl"
refers to a non-aromatic cyclic hydrocarbon ring having from three
to eight carbon atoms. Exemplary "C.sub.3-C.sub.8 cycloalkyl"
groups useful in the present invention include, but are not limited
to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl
and cyclooctyl.
The term "C.sub.5-C.sub.8cycloalkenyl" refers to a non-aromatic
monocyclic carboxycyclic ring having the specified number of carbon
atoms and up to 3 carbon-carbon double bonds. "Cycloalkenyl"
includes by way of example cyclopentenyl and cyclohexenyl.
Where "C.sub.3-C.sub.8 heterocycloalkyl" is used, it means a
non-aromatic heterocyclic ring containing the specified number of
ring atoms being, saturated or having one or more degrees of
unsaturation and containing one or more heteroatom substitutions
selected from O, S and/or N. Such a ring may be optionally fused to
one or more other "heterocyclic" ring(s) or cycloalkyl ring(s).
Examples of "heterocyclic" moieties include, but are not limited
to, aziridine, thiirane, oxirane, azetidine, oxetane, thietane,
tetrahydrofuran, pyran, 1,4-dioxane, 1,3-dioxane, piperidine,
piperazine, 2,4-piperazinedione, pyrrolidine, imidazolidine,
pyrazolidine, morpholine, thiomorpholine, tetrahydrothiopyran,
tetrahydrothiophene, and the like.
"Aryl" refers to optionally substituted monocyclic and
polycarbocyclic unfused or fused groups having 6 to 14 carbon atoms
and having at least one aromatic ring that complies with Huckel's
Rule. Examples of aryl groups are phenyl, biphenyl, naphthyl,
anthracenyl, phenanthrenyl and the like.
"Heteroaryl" means an optionally substituted aromatic monocyclic
ring or polycarbocyclic fused ring system wherein at least one ring
complies with Huckel's Rule, has the specified number of ring
atoms, and that ring contains at least one heteratom selected from
N, O, and/or S. Examples of "heteroaryl" groups include furanyl,
thiophenyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl,
thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, oxo-pyridyl,
thiadiazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrazinyl,
pyrimidinyl, quinolinyl, isoquinolinyl, benzofuranyl,
benzothiophenyl, indolyl, and indazolyl.
The term "optionally" means that the subsequently described
event(s) may or may not occur, and includes both event(s), which
occur, and events that do not occur.
The term "solvate" refers to a complex of variable stoichiometry
formed by a solute and a solvent. Such solvents for the purpose of
the invention may not interfere with the biological activity of the
solute. Examples of suitable solvents include, but are not limited
to, water, methanol, ethanol and acetic acid. Preferably the
solvent used is a pharmaceutically acceptable solvent. Examples of
suitable pharmaceutically acceptable solvents include, without
limitation, water, ethanol and acetic acid. Most preferably the
solvent used is water.
Herein, the term "pharmaceutically-acceptable salts" refers to
salts that retain the desired biological activity of the subject
compound and exhibit minimal undesired toxicological effects. These
pharmaceutically-acceptable salts may be prepared in situ during
the final isolation and purification of the compound, or by
separately reacting the purified compound in its free acid or free
base form with a suitable base or acid, respectively.
In certain embodiments, compounds according to Formula I may
contain an acidic functional group, one acidic enough to form
salts. Representative salts include pharmaceutically-acceptable
metal salts such as sodium, potassium, lithium, calcium, magnesium,
aluminum, and zinc salts; carbonates and bicarbonates of a
pharmaceutically-acceptable metal cation such as sodium, potassium,
lithium, calcium, magnesium, aluminum, and zinc;
pharmaceutically-acceptable organic primary, secondary, and
tertiary amines including aliphatic amines, aromatic amines,
aliphatic diamines, and hydroxy alkylamines such as methylamine,
ethylamine, 2-hydroxy-ethylamine, diethylamine, triethylamine,
ethylenediamine, ethanolamine, diethanolamine, and
cyclohexylamine.
In certain embodiments, compounds according to Formula (I) may
contain a basic functional group and are therefore capable of
forming pharmaceutically-acceptable acid addition salts by
treatment with a suitable acid. Suitable acids include
pharmaceutically-acceptable inorganic acids and
pharmaceutically-acceptable organic acids. Representative
pharmaceutically-acceptable acid addition salts include
hydrochloride, hydrobromide, nitrate, methylnitrate, sulfate,
bisulfate, sulfamate, phosphate, acetate, hydroxyacetate,
phenylacetate, propionate, butyrate, isobutyrate, valerate,
maleate, hydroxymaleate, acrylate, fumarate, malate, tartrate,
citrate, salicylate, p-aminosalicyclate, glycollate, lactate,
heptanoate, phthalate, oxalate, succinate, benzoate,
o-acetoxybenzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate,
hydroxybenzoate, methoxybenzoate, mandelate, tannate, formate,
stearate, ascorbate, palmitate, oleate, pyruvate, pamoate,
malonate, laurate, glutarate, glutamate, estolate, methanesulfonate
(mesylate), ethanesulfonate (esylate), 2-hydroxyethanesulfonate,
benzenesulfonate (besylate), p-aminobenzenesulfonate,
p-toluenesulfonate (tosylate), and napthalene-2-sulfonate.
A group of compounds of particular interest are those wherein:
R.sup.1 is selected from the group consisting of hydrogen,
C.sub.1-C.sub.10alkyl, C.sub.2-C.sub.10alkenyl,
C.sub.2-C.sub.10alkynyl, C.sub.3-C.sub.8cycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8cycloalkyl,
C.sub.5-C.sub.8cycloalkenyl, C.sub.1-C.sub.10alkyl-C.sub.5-C.sub.8
cycloalkenyl, C.sub.3-C.sub.8 heterocycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8 heterocycloalkyl, aryl,
C.sub.1-C.sub.10alkyl-aryl, heteroaryl and
C.sub.1-C.sub.10alkyl-heteroaryl;
R.sup.4 is selected from the group consisting of hydrogen,
COOR.sup.9, CONR.sup.7R.sup.8, --NR.sup.5R.sup.6,
C.sub.1-C.sub.10alkyl, C.sub.2-C.sub.10alkenyl,
C.sub.2-C.sub.10alkynyl, C.sub.3-C.sub.8cycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8cycloalkyl,
C.sub.5-C.sub.8cycloalkenyl, C.sub.1-C.sub.10alkyl-C.sub.5-C.sub.8
cycloalkenyl, C.sub.3-C.sub.8 heterocycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8 heterocycloalkyl, aryl,
C.sub.1-C.sub.10alkyl-aryl, heteroaryl and
C.sub.1-C.sub.10alkyl-heteroaryl;
R.sup.2 is --NR.sup.7R.sup.8, --OR.sup.9;
R.sup.3 is H or C.sub.1-C.sub.4alkyl;
R.sup.9 is H or a cation, or C.sub.1-C.sub.10alkyl which is
unsubstituted or substituted with one or more substituents
independently selected from the group consisting of C.sub.3-C.sub.6
cycloalkyl, heterocycloalkyl, aryl, and heteroaryl; and wherein any
carbon or heteroatom of R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.9
is unsubstituted or, where possible, is substituted with one or
more substituents independently selected from the group consisting
of C.sub.1-C.sub.6 alkyl, aryl, heteroaryl, halogen, --OR.sup.10,
--NR.sup.5R.sup.6, cyano, nitro, --C(O)R.sup.10, --C(O)OR.sup.10,
--SR.sup.10, --S(O)R.sup.10, --S(O).sub.2R.sup.10,
--NR.sup.5R.sup.6, --CONR.sup.5R.sup.6, --N(R.sup.5)C(O)R.sup.10,
--N(R.sup.5)C (O)OR.sup.10, OC(O)NR.sup.5R.sup.6,
N(R.sup.5)C(O)NR.sup.5R.sup.6, --SO.sub.2NR.sup.5R.sup.6,
--N(R.sup.5)SO.sub.2R.sup.10, C.sub.1-C.sub.10 alkenyl,
C.sub.1-C.sub.10 alkynyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.3-C.sub.6 heterocycloalkyl, aryl and heteroaryl group,
wherein R.sup.5, and R.sup.6 are the same as defined above and
R.sup.10 is hydrogen, C.sub.1-C.sub.10alkyl,
C.sub.2-C.sub.10alkenyl, C.sub.2-C.sub.10alkynyl,
--CO(C.sub.1-C.sub.4 alkyl), --CO(aryl), --CO(heteroaryl),
--CO(C.sub.3-C.sub.6 cycloalkyl), --CO(C.sub.3-C.sub.6
heterocycloalkyl), --SO.sub.2(C.sub.1-C.sub.4 alkyl),
C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8heterocycloalkyl,
C.sub.6-C.sub.14 aryl, C.sub.1-C.sub.10alkyl-aryl, heteroaryl, or
C.sub.1-C.sub.10alkyl-heteroaryl; or a pharmaceutically acceptable
salt or solvate thereof.
Another group of compounds of particular interest are those
wherein:
R.sup.1 is selected from the group consisting of hydrogen,
C.sub.1-C.sub.10alkyl, C.sub.2-C.sub.10alkenyl,
C.sub.2-C.sub.10alkynyl, C.sub.3-C.sub.8cycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8cycloalkyl,
C.sub.5-C.sub.8cycloalkenyl,
C.sub.1-C.sub.10alkyl-C.sub.5-C.sub.8cycloalkenyl, C.sub.3-C.sub.8
heterocycloalkyl, C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8
heterocycloalkyl, aryl, C.sub.1-C.sub.10alkyl-aryl, heteroaryl and
C.sub.1-C.sub.10alkyl-heteroaryl;
R.sup.4 is selected from the group consisting of hydrogen,
COOR.sup.9, CONR.sup.7R.sup.8, --NR.sup.5R.sup.6,
C.sub.1-C.sub.10alkyl, C.sub.2-C.sub.10alkenyl,
C.sub.2-C.sub.10alkynyl, C.sub.3-C.sub.8cycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8cycloalkyl,
C.sub.5-C.sub.8cycloalkenyl, C.sub.1-C.sub.10alkyl-C.sub.5-C.sub.8
cycloalkenyl, C.sub.3-C.sub.8 heterocycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8 heterocycloalkyl, aryl,
C.sub.1-C.sub.10alkyl-aryl, heteroaryl and
C.sub.1-C.sub.10alkyl-heteroaryl;
R.sup.2 is --NR.sup.7R.sup.8, --OR.sup.9;
R.sup.3 is H;
R.sup.9 is H or a cation;
wherein any carbon or heteroatom of R.sup.1, R.sup.2, R.sup.3,
R.sup.4 is unsubstituted or, where possible, is substituted with
one or more substituents independently selected from
C.sub.1-C.sub.6 alkyl, aryl, heteroaryl, halogen, --OR.sup.10,
--NR.sup.5R.sup.6, cyano, nitro, --C(O)R.sup.10, --C(O)OR.sup.10,
--SR.sup.10, --S(O)R.sup.10, --S(O).sub.2R.sup.10,
--NR.sup.5R.sup.6, --CONR.sup.5R.sup.6, --N(R.sup.5)C(O)R.sup.10,
--N(R.sup.5)C(O) OR.sup.10, --OC(O)NR.sup.5R.sup.6,
--N(R.sup.5)C(O)NR.sup.5R.sup.6, --SO.sub.2NR.sup.5R.sup.6,
--N(R.sup.5)SO.sub.2R.sup.10, C.sub.1-C.sub.10 alkenyl,
C.sub.1-C.sub.10 alkynyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.3-C.sub.6 heterocycloalkyl, aryl or heteroaryl group, wherein
R.sup.5, and R.sup.6 are the same as defined above and R.sup.10 is
hydrogen, C.sub.1-C.sub.10alkyl, C.sub.2-C.sub.10alkenyl,
C.sub.2-C.sub.10alkynyl, --CO(C.sub.1-C.sub.4 alkyl), --CO(aryl),
--CO(heteroaryl), --CO(C.sub.3-C.sub.6 cycloalkyl),
--CO(C.sub.3-C.sub.6 heterocycloalkyl), --SO.sub.2(C.sub.1-C.sub.4
alkyl), C.sub.3-C.sub.8 cycloalkyl,
C.sub.3-C.sub.8heterocycloalkyl, C.sub.6-C.sub.14 aryl,
C.sub.1-C.sub.10alkyl-aryl, heteroaryl, and
C.sub.1-C.sub.10alkyl-heteroaryl;
or a pharmaceutically acceptable salt thereof.
Another group of compounds of particular interest are those
wherein:
R.sup.1 is selected from the group consisting of hydrogen,
C.sub.1-C.sub.10alkyl, C.sub.2-C.sub.10alkenyl,
C.sub.2-C.sub.10alkynyl, C.sub.3-C.sub.8cycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8cycloalkyl,
C.sub.5-C.sub.8cycloalkenyl, C.sub.1-C.sub.10alkyl-C.sub.5-C.sub.8
cycloalkenyl, C.sub.3-C.sub.8 heterocycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8 heterocycloalkyl, aryl,
C.sub.1-C.sub.10alkyl-aryl, heteroaryl and
C.sub.1-C.sub.10alkyl-heteroaryl;
R.sup.4 is selected from the group consisting of hydrogen,
COOR.sup.9, CONR.sup.7R.sup.8, --NR.sup.5R.sup.6,
C.sub.1-C.sub.10alkyl, C.sub.2-C.sub.10alkenyl,
C.sub.2-C.sub.10alkynyl, C.sub.3-C.sub.8cycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8cycloalkyl,
C.sub.5-C.sub.8cycloalkenyl, C.sub.1-C.sub.10alkyl-C.sub.5-C.sub.8
cycloalkenyl, C.sub.3-C.sub.8 heterocycloalkyl,
C.sub.1-C.sub.10alkyl-C.sub.3-C.sub.8 heterocycloalkyl, aryl,
C.sub.1-C.sub.10alkyl-aryl, heteroaryl and
C.sub.1-C.sub.10alkyl-heteroaryl;
R.sup.2 is --OR.sup.9;
R.sup.3 is H;
R.sup.9 is H or a cation;
wherein any carbon or heteroatom of R.sup.1, R.sup.4 is
unsubstituted or, where possible, is substituted with one or more
substituents independently selected from C.sub.1-C.sub.6 alkyl,
aryl, heteroaryl, halogen, --OR.sup.10, --NR.sup.5R.sup.6, cyano,
nitro, --C(O) R.sup.10, --C(O)OR.sup.10, --SR.sup.10,
--S(O)R.sup.10, --S(O).sub.2R.sup.10, --NR.sup.5R.sup.6,
--CONR.sup.5R.sup.6, --N(R.sup.5)C(O)R.sup.10, --N(R.sup.5)C(O)
OR.sup.10, --OC(O)NR.sup.5R.sup.6, --N(R.sup.5)C(O)NR.sup.5R.sup.6,
--SO.sub.2NR.sup.5R.sup.6, --N(R.sup.5)SO.sub.2R.sup.10,
C.sub.1-C.sub.10 alkenyl, C.sub.1-C.sub.10 alkynyl, C.sub.3-C.sub.6
cycloalkyl, C.sub.3-C.sub.6 heterocycloalkyl, aryl or heteroaryl
group, wherein R.sup.5, and R.sup.6 are the same as defined above
and R.sup.10 is hydrogen, C.sub.1-C.sub.10alkyl,
C.sub.2-C.sub.10alkenyl, C.sub.2-C.sub.10alkynyl,
--CO(C.sub.1-C.sub.4 alkyl), --CO(aryl), --CO(heteroaryl),
--CO(C.sub.3-C.sub.6 cycloalkyl), --CO(C.sub.3-C.sub.6
heterocycloalkyl), --SO.sub.2(C.sub.1-C.sub.4 alkyl),
C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8heterocycloalkyl,
C.sub.6-C.sub.14 aryl, C.sub.1-C.sub.10alkyl-aryl, heteroaryl, and
C.sub.1-C.sub.10alkyl-heteroaryl;
or a pharmaceutically acceptable salt thereof.
Compounds of further interest are those wherein:
R.sup.1 is 2-fluoro-4-bromobenzyl, 4-bromobenzyl,
4-(halophenyl)benzyl, 2-fluoro-4-(halophenyl)benzyl,
2-fluoro-4-triflurormethylbenzyl, 4-trifluorobenzyl, or
2-fluoro-4-(C.sub.1-C.sub.4alkoxyphenyl)benzyl;
R.sup.2 is OH;
R.sup.3 is H;
R.sup.4 is isopropyl, t-butyl or cyclohexyl; or
a pharmaceutically acceptable salt thereof.
Processes for preparing the compound of formula (I) are also within
the ambit of this invention. To illustrate, a process for preparing
a compound of formula (I)
##STR00003## wherein R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are the
same as defined above for formula (I), the process comprising
treating a compound of formula A:
##STR00004## wherein R.sup.1 and R.sup.4 are the same as for those
groups in formula (I) with an .alpha.-aminoacid sodium salt in an
appropriate solvent, such as 2-methoxyethanol, under either
conventional thermal conditions or by microwave irradiation, to
form a compound of formula (I) where R.sup.2 is --OH;
It will be appreciated by those skilled in the art that the
compounds of formula (I) may exist in one or more tautomeric forms
such as:
##STR00005##
All tautomeric forms of the compounds described herein, including
mixtures thereof, are intended to be encompassed within the scope
of the invention. Generally, the compounds exemplified herein have
been assigned names based on the structure of the tautomer of
formula (IA). It should be understood that any reference to named
compounds of this invention is intended to encompass all tautomers
of the named compounds and any mixtures of tautomers of the named
compounds.
The compounds of formula (I) may be prepared in crystalline or
non-crystalline form, and, if crystalline, may optionally be
solvated, e.g. as the hydrate. This invention includes within its
scope stoichiometric solvates (e.g. hydrates) as well as compounds
containing variable amounts of solvent (e.g. water).
Certain of the compounds described herein may contain one or more
chiral atoms, or may otherwise be capable of existing as two
enantiomers. The compounds claimed below include mixtures of
enantiomers as well as purified enantiomers or enantiomerically
enriched mixtures. Also included within the scope of the invention
are the individual isomers of the compounds represented by formula
(I), or claimed below, as well as any wholly or partially
equilibrated mixtures thereof. The present invention also covers
the individual isomers of the claimed compounds as mixtures with
isomers thereof in which one or more chiral centers are inverted.
Also, it is understood that any tautomers and mixtures of tautomers
of the claimed compounds are included within the scope of the
compounds of formula (I) as disclosed herein above or claimed
herein below.
Where there are different isomeric forms they may be separated or
resolved one from the other by conventional methods, or any given
isomer may be obtained by conventional synthetic methods or by
stereospecific or asymmetric syntheses.
While it is possible that, for use in therapy, a compound of
formula (I), as well as salts, solvates and the like, may be
administered as a neat preparation, i.e. no additional carrier, the
more usual practice is to present the active ingredient confected
with a carrier or diluent. Accordingly, the invention further
provides pharmaceutical compositions, which includes a compound of
formula (I) and salts, solvates and the like, and one or more
pharmaceutically acceptable carriers, diluents, or excipients. The
compounds of formula (I) and salts, solvates, etc, are as described
above. The carrier(s), diluent(s) or excipient(s) must be
acceptable in the sense of being compatible with the other
ingredients of the formulation and not deleterious to the recipient
thereof. In accordance with another aspect of the invention there
is also provided a process for the preparation of a pharmaceutical
formulation including admixing a compound of the formula (I), or
salts, solvates etc, with one or more pharmaceutically acceptable
carriers, diluents or excipients.
It will be appreciated by those skilled in the art that certain
protected derivatives of compounds of formula (I), which may be
made prior to a final deprotection stage, may not possess
pharmacological activity as such, but may, in certain instances, be
administered orally or parenterally and thereafter metabolised in
the body to form compounds of the invention which are
pharmacologically active. Such derivatives may therefore be
described as "prodrugs". Further, certain compounds of the
invention may act as prodrugs of other compounds of the invention.
All protected derivatives and prodrugs of compounds of the
invention are included within the scope of the invention. Examples
of suitable pro-drugs for the compounds of the present invention
are described in Drugs of Today, Volume 19, Number 9, 1983, pp
499-538 and in Topics in Chemistry, Chapter 31, pp 306-316 and in
"Design of Prodrugs" by H. Bundgaard, Elsevier, 1985, Chapter 1
(the disclosures in which documents are incorporated herein by
reference). It will further be appreciated by those skilled in the
art, that certain moieties, known to those skilled in the art as
"pro-moieties", for example as described by H. Bundgaard in "Design
of Prodrugs" (the disclosure in which document is incorporated
herein by reference) may be placed on appropriate functionalities
when such functionalities are present within compounds of the
invention. Preferred prodrugs for compounds of the invention
include: esters, carbonate esters, hemi-esters, phosphate esters,
nitro esters, sulfate esters, sulfoxides, amides, carbamates,
azo-compounds, phosphamides, glycosides, ethers, acetals and
ketals.
Pharmaceutical compositions may be presented in unit dose forms
containing a predetermined amount of active ingredient per unit
dose. Such a unit may contain, for example, 0.5 mg to 1 g,
preferably 1 mg to 700 mg, more preferably 5 mg to 100 mg of a
compound of the formula (I), depending on the condition being
treated, the route of administration and the age, weight and
condition of the patient, or pharmaceutical compositions may be
presented in unit dose forms containing a predetermined amount of
active ingredient per unit dose. Preferred unit dosage compositions
are those containing a daily dose or sub-dose, as herein above
recited, or an appropriate fraction thereof, of an active
ingredient. Furthermore, such pharmaceutical compositions may be
prepared by any of the methods well known in the pharmacy art.
Pharmaceutical compositions may be adapted for administration by
any appropriate route, for example by the oral (including buccal or
sublingual), rectal, nasal, topical (including buccal, sublingual
or transdermal), vaginal or parenteral (including subcutaneous,
intramuscular, intravenous or intradermal) route. Such compositions
may be prepared by any method known in the art of pharmacy, for
example by bringing into association a compound of formal (I) with
the carrier(s) or excipient(s).
Pharmaceutical compositions adapted for oral administration may be
presented as discrete units such as capsules or tablets; powders or
granules; solutions or suspensions in aqueous or non-aqueous
liquids; edible foams or whips; or oil-in-water liquid emulsions or
water-in-oil liquid emulsions.
Capsules are made by preparing a powder mixture, as described
above, and filling formed gelatin sheaths. Glidants and lubricants
such as colloidal silica, talc, magnesium stearate, calcium
stearate or solid polyethylene glycol can be added to the powder
mixture before the filling operation. A disintegrating or
solubilizing agent such as agar-agar, calcium carbonate or sodium
carbonate can also be added to improve the availability of the
medicament when the capsule is ingested.
Moreover, when desired or necessary, suitable binders, lubricants,
disintegrating agents and coloring agents can also be incorporated
into the mixture. Suitable binders include starch, gelatin, natural
sugars such as glucose or beta-lactose, corn sweeteners, natural
and synthetic gums such as acacia, tragacanth or sodium alginate,
carboxymethylcellulose, polyethylene glycol, waxes and the like.
Lubricants used in these dosage forms include sodium oleate, sodium
stearate, magnesium stearate, sodium benzoate, sodium acetate,
sodium chloride and the like. Disintegrators include, without
limitation, starch, methyl cellulose, agar, bentonite, xanthan gum
and the like. Tablets are formulated, for example, by preparing a
powder mixture, granulating or slugging, adding a lubricant and
disintegrant and pressing into tablets. A powder mixture is
prepared by mixing the compound, suitably comminuted, with a
diluent or base as described above, and optionally, with a binder
such as carboxymethylcellulose, an aliginate, gelatin, or polyvinyl
pyrrolidone, a solution retardant such as paraffin, a resorption
accelerator such as a quaternary salt and/or an absorption agent
such as bentonite, kaolin or dicalcium phosphate. The powder
mixture can be granulated by tablet forming dies by means of the
addition of stearic acid, a stearate salt, talc or mineral oil. The
lubricated mixture is then compressed into tablets. The compounds
of the present invention can also be combined with a free flowing
inert carrier and compressed into tablets directly without going
through the granulating or slugging steps. A clear or opaque
protective coating consisting of a sealing coat of shellac, a
coating of sugar or polymeric material and a polish coating of wax
can be provided. Dyestuffs can be added to these coatings to
distinguish different unit dosages.
Oral fluids such as solution, syrups and elixirs can be prepared in
dosage unit form so that a given quantity contains a predetermined
amount of a compound of formula (I). Syrups can be prepared by
dissolving the compound in a suitably flavored aqueous solution,
while elixirs are prepared through the use of a non-toxic alcoholic
vehicle. Suspensions can be formulated by dispersing the compound
in a non-toxic vehicle. Solubilizers and emulsifiers such as
ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol
ethers, preservatives, flavor additive such as peppermint oil or
natural sweeteners or saccharin or other artificial sweeteners, and
the like can also be added.
Where appropriate, dosage unit pharmaceutical compositions for oral
administration can be microencapsulated. The formulation can also
be prepared to prolong or sustain the release as for example by
coating or embedding particulate material in polymers, wax or the
like.
Pharmaceutical compositions adapted for rectal administration may
be presented as suppositories or as enemas.
Pharmaceutical compositions adapted for vaginal administration may
be presented as pessaries, tampons, creams, gels, pastes, foams or
spray formulations.
Pharmaceutical formulations adapted for parenteral administration
include aqueous and non-aqueous sterile injection solutions which
may contain anti-oxidants, buffers, bacteriostats and solutes which
render the composition isotonic with the blood of the intended
recipient; and aqueous and non-aqueous sterile suspensions which
may include suspending agents and thickening agents. The
pharmaceutical compositions may be presented in unit-dose or
multi-dose containers, for example sealed ampoules and vials, and
may be stored in a freeze-dried (lyophilized) condition requiring
only the addition of the sterile liquid carrier, for example water
for injections, immediately prior to use. Extemporaneous injection
solutions and suspensions may be prepared from sterile powders,
granules and tablets.
It should be understood that in addition to the ingredients
particularly mentioned above, the pharmaceutical compositions may
include other agents conventional in the art having regard to the
type of formulation in question, for example those suitable for
oral administration may include flavouring agents.
A therapeutically effective amount of a compound of the present
invention will depend upon a number of factors including, for
example, the age and weight of the intended recipient, the precise
condition requiring treatment and its severity, the nature of the
formulation, and the route of administration, and will ultimately
be at the discretion of the attendant prescribing the medication.
However, an effective amount of a compound of formula (I) for the
treatment of anemia will generally be in the range of 0.1 to 100
mg/kg body weight of recipient per day and more usually in the
range of 1 to 10 mg/kg body weight per day. Thus, for a 70 kg adult
mammal, the actual amount per day would usually be from 70 to 700
mg and this amount may be given in a single dose per day or more
usually in a number (such as two, three, four, five or six) of
sub-doses per day such that the total daily dose is the same. An
effective amount of a salt or solvate, etc., may be determined as a
proportion of the effective amount of the compound of formula (I)
per se. It is envisaged that similar dosages would be appropriate
for treatment of the other conditions referred to above.
DEFINITIONS
rt--room temperature
DBU--1,8-diazabicyclo[5.4.0]undec-7-ene
DCM--dichloromethane
DMF--dimethylformamide
DMSO--dimethylsulfoxide
KHMDS--potassium hexamethyldisilazide
LCMS--liquid chromatography/mass spectrometry
MTBE--methyl t-butyl ether
NMR--nuclear magnetic resonance
ODS--octadecyl silyl
PTFE--polytetrafluoroethylene
RP-HPLC--reverse-phase high performance liquid chromatography
TFA--Trifluoroacetic acid
THF--tetrahydrofuran
Chemical Background:
The compounds of this invention may be made by a variety of
methods, including standard chemistry. Any previously defined
variable will continue to have the previously defined meaning
unless otherwise indicated. Illustrative general synthetic methods
are set out below and then specific compounds of the invention as
prepared are given in the examples.
Compounds of general formula (I) may be prepared by methods known
in the art of organic synthesis as set forth in part by the
following synthesis schemes. In all of the schemes described below,
it is well understood that protecting groups for sensitive or
reactive groups are employed where necessary in accordance with
general principles of chemistry. Protecting groups are manipulated
according to standard methods of organic synthesis (T. W. Green and
P. G. M. Wuts (1991) Protecting Groups in Organic Synthesis, John
Wiley & Sons). These groups are removed at a convenient stage
of the compound synthesis using methods that are readily apparent
to those skilled in the art. The selection of processes as well as
the reaction conditions and order of their execution shall be
consistent with the preparation of compounds of formula (I). Those
skilled in the art will recognize if a stereocenter exists in
compounds of formula (I). Accordingly, the present invention
includes both possible stereoisomers and includes not only racemic
compounds but the individual enantiomers as well. When a compound
is desired as a single enantiomer, it may be obtained by
stereospecific synthesis or by resolution of the final product or
any convenient intermediate. Resolution of the final product, an
intermediate, or a starting material may be effected by any
suitable method known in the art. See, for example, Stereochemistry
of Organic Compounds by E. L. Eliel, S. H. Wilen, and L. N. Mander
(Wiley-Interscience, 1994).
Illustrated Methods of Preparation
##STR00006## a) R.sup.1NHNH.sub.2.2HCl, EtNPr.sup.i.sub.2 or NaOAc,
CH.sub.2Cl.sub.2 or R.sup.1NHNH.sub.2.2HCl, DBU, EtOH, microwave,
150.degree. C.; b) ClOCCH.sub.2CO.sub.2Et, NaH or DBU, THF, rt or
60.degree. C.; c) DBU, THF or dioxane, reflux or microwave,
130.degree. C.; d) NaO.sub.2CCH.sub.2NH.sub.2,
MeOCH.sub.2CH.sub.2OH, reflux.
##STR00007## a) EtO.sub.2CCH.sub.2CONHNH.sub.2, AcOH,
CH.sub.2Cl.sub.2 or EtO.sub.2CCH.sub.2CONHNH.sub.2, TsOH, PhMe,
reflux or EtO.sub.2CCH.sub.2CONHNH.sub.2, AcOH, EtOH, (with or
without MgSO.sub.4), reflux or microwave, 150.degree. C.; b) KHMDS,
KOBu.sup.t or DBU, PhMe, Bu.sup.tOH or dioxane, reflux or
microwave, 150.degree. C. or KOBu.sup.t, Bu.sup.tOH, microwave,
150.degree. C.; c) NaH, R.sup.1Br, DMF, 0.degree. C. to rt; d)
NaO.sub.2CCH.sub.2NH.sub.2, MeOCH.sub.2CH.sub.2OH or EtOH, reflux
or microwave, 150.degree. C.
##STR00008## a) R.sup.4MgBr or R.sup.4Li, THF or Et.sub.2O,
-78.degree. C. to 0.degree. C.
##STR00009## a) 48% aq HBr, AcOH, reflux
##STR00010## a) ArB(OH).sub.2 or ArB(OR).sub.2,
Pd(PPh.sub.3).sub.4, aq K.sub.2CO.sub.3, dioxane, microwave,
100.degree. C.
##STR00011## a) KHMDS, dioxane, reflux; b) NaH, R.sup.1Br, DMF,
0.degree. C. to rt; c) NaO.sub.2CCH.sub.2NH.sub.2.
MeOCH.sub.2CH.sub.2OH, reflux or microwave, 150.degree. C.g
EXPERIMENTALS
Example 1
##STR00012##
N-{[5-Hydroxy-6-methyl-3-oxo-2-(phenylmethyl)-2,3-dihydro-4-pyridazinyl]ca-
rbonyl}glycine
1a) Ethyl
3-[2-[2-(ethyloxy)-1-methyl-2-oxoethylidene]-1-(phenylmethyl)hy-
drazino]-3-oxopropanoate. Ethyl pyruvate (0.324 g, 2.79 mmol) was
added to a stirred mixture of benzylhydrazine dihydrochloride
(0.551 g, 2.82 mmol), diisopropylethylamine (1.00 mL, 5.74 mmol)
and dichloromethane (10 mL). Magnesium sulfate (excess) was added
and the mixture stirred 0.5 h, then loaded onto a short column of
silica gel and the product eluted (50% ethyl acetate/hexane). After
evaporation of the solvent, the crude hydrazone (0.339 g) was
dissolved in tetrahydrofuran (20 mL) and sodium hydride (0.075 g of
a 60% oil suspension, 1.88 mmol) added with stirring. The mixture
was stirred 20 min, then ethyl 3-chloro-3-oxopropionate (0.235 mL,
1.84 mmol) injected dropwise and stirring continued at room
temperature 10 min and at 60.degree. C. for 0.5 h. Further sodium
hydride (0.075 g of a 60% oil suspension, 1.88 mmol) was added and
stirring continued at 65.degree. C. for 0.5 h. The mixture was
cooled, poured into 0.1M aqueous hydrochloric acid (100 mL) and
extracted with ethyl acetate. The extracts were dried (MgSO.sub.4)
and evaporated under reduced pressure. Ethyl pyruvate (0.324 g,
2.79 mmol) was added to a stirred mixture of the residue in
dichloromethane (10 mL) and magnesium sulfate. After 2 h, the
mixture was chromatographed (silica gel, 20-50% ethyl
acetate/hexane) to give the title compound (0.205 g, 22%) as a gum.
LCMS (ES.sup.+) m/z 335 (MH.sup.+).
1b) Ethyl
5-hydroxy-6-methyl-3-oxo-2-(phenylmethyl)-2,3-dihydro-4-pyridaz-
inecarboxylate. 1,8-Diazabicyclo[5.4.0]undec-7-ene (0.180 mL, 1.20
mmol) was added to a stirred solution of the compound from example
1(a) (0.203 g, 0.607 mmol) in tetrahydrofuran (5 mL) in 2 portions
while heating under reflux under nitrogen over 2 h. The mixture was
cooled, poured into 0.1M aqueous hydrochloric acid (50 mL) and
extracted with ethyl acetate. The extracts were dried (MgSO.sub.4),
evaporated under reduced pressure and the residue chromatographed
(silica gel, 1-5% methanol/dichloromethane) to give the title
compound (0.128 g, 73%) as a solid. 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 1.26 (t, J=7.20 Hz, 3H) 2.21 (s, 3H) 4.26 (q, J=7.16
Hz, 2H) 5.14 (s, 2 H) 7.24-7.36 (m, 5H) 12.13 (s, 1H).
1c)
N-{[5-Hydroxy-6-methyl-3-oxo-2-(phenylmethyl)-2,3-dihydro-4-pyridazin-
yl]carbonyl}glycine. A stirred mixture of the compound from example
1(b) (0.126 g, 0.437 mmol), anhydrous glycine, sodium salt (0.090
g, 0.927 mmol) and 2-methoxyethanol (4 mL) was heated under reflux
under nitrogen for 2 h, cooled and diluted with water (30 mL). 1M
aqueous hydrochloric acid (1 mL) was added slowly and the mixture
stirred 2 h, then the solid filtered, washed with water and dried
to leave the title compound (0.104 g, 75%) as a tan powder. 1H NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 2.25 (s, 3H) 4.10 (d, J=5.56
Hz, 2H) 5.25 (s, 2H) 7.27-7.37 (m, 5H) 10.17 (t, J=5.05 Hz, 1H)
12.97 (s, 1H) 15.65 (s, 1H).
Example 2
##STR00013##
N-{[5-Hydroxy-3-oxo-6-phenyl-2-(phenylmethyl)-2,3-dihydro-4-pyridazinyl]ca-
rbonyl}glycine
2a) Ethyl
3-[2-[2-(ethyloxy)-2-oxo-l-phenylethylidene]-1-(phenylmethyl)hy-
drazino]-3-oxopropanoate. A mixture of ethyl oxo(phenyl)acetate
(0.315 g, 1.77 mmol), benzylhydrazine dihydrochloride (0.334 g,
1.71 mmol), diisopropylethylamine (0.596 mL, 3.42 mmol),
dichloromethane (10 mL) and excess magnesium sulfate was stirred
for 18 h, then filtered through a plug of silica gel and the cake
washed with 50% ethyl acetate/hexane. The solvent was removed under
reduced pressure and the crude hydrazone dissolved in
tetrahydrofuran (5 mL) and placed under nitrogen.
1,8-Diazabicyclo[5.4.0]undec-7-ene (0.212 mL, 1.42 mmol) was added,
followed by ethyl 3-chloro-3-oxopropionate (0.182 mL, 1.42 mmol)
dropwise. The mixture was stirred at room temperature for 2 h, then
poured into water (50 mL) and extracted with ethyl acetate. The
extracts were dried (MgSO.sub.4), evaporated under reduced pressure
and the residue chromatographed (silica gel, 10-50% ethyl
acetate/hexane) to give the title compound (0.117 g) as a gum,
sufficiently pure to use in the next reaction. LCMS (ES.sup.+) m/z
397 (MH.sup.+).
2b) Ethyl
5-hydroxy-3-oxo-6-phenyl-2-(phenylmethyl)-2,3-dihydro-4-pyridaz-
inecarboxylate. 1,8-Diazabicyclo[5.4.0]undec-7-ene (0.100 mL, 0.670
mmol) was added to a stirred solution of the compound from example
2(a) (0.115 g, 0.290 mmol) in tetrahydrofuran (5 mL) under nitrogen
and the mixture stirred under reflux for 2 h, then cooled, poured
into 0.1M aqueous hydrochloric acid (50 mL) and extracted with
ethyl acetate. The extracts were dried (MgSO.sub.4), evaporated
under reduced pressure and the residue chromatographed (silica gel,
1-5% methanol/dichloromethane). The material obtained was
chromatographed again (silica gel, 20-30% ethyl acetate/hexane,
then 1-4% methanol/dichloromethane) to give the title compound
(0.037 g, 6% over 3 steps) as a solid. 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 1.28 (t, J=7.08 Hz, 3H) 4.29 (q, J=7.16
Hz, 2H) 5.25 (s, 2H) 7.27-7.38 (m, 5H) 7.44-7.49 (m, 3H), 7.66-7.71
(m, 2H).
2c)
N-{[5-Hydroxy-3-oxo-6-phenyl-2-(phenylmethyl)-2,3-dihydro-4-pyridazin-
yl]carbonyl}glycine. A stirred mixture of the compound from example
2(b) (0.036 g, 0.103 mmol), anhydrous glycine, sodium salt (0.050
g, 0.515 mmol) and 2-methoxyethanol (2 mL) was heated under reflux
under nitrogen for 2 h, cooled and diluted with water (20 mL). 1M
aqueous hydrochloric acid (0.5 mL) was added slowly and the mixture
allowed to stand for 18 h, then the solid filtered, washed with
water and dried to leave the title compound (0.030 g, 77%) as a
pale pink solid. 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 4.13
(d, J=5.56 Hz, 2H) 5.37 (s, 2 H) 7.29-7.39 (m, 5H) 7.46-7.54 (m,
3H) 7.75-7.82 (m, 2H) 10.26 (t, J=4.93 Hz, 1H) 13.00 (br s,
1H).
Example 3
##STR00014##
N-{[5-Hydroxy-6-(1-methylethyl)-3-oxo-2-(phenylmethyl)-2,3-dihydro-4-pyrid-
azinyl]carbonyl}glycine
3a) Ethyl 3-oxo-3-[1-(phenylmethyl)hydrazino]propanoate. A mixture
of benzylhydrazine dihydrochloride (2.50 g, 12.8 mmol), potassium
carbonate (1.77 g, 12.8 mmol), acetone (6.0 mL, 82 mmol) and ethyl
acetate (30 mL) was stirred at room temperature while a solution of
1M aqueous sodium hydroxide (6.0 mL, 6.0 mmol) was added dropwise.
After the addition, the mixture was stirred 0.5 h, then excess
magnesium sulfate added and the mixture filtered. The filtrate was
evaporated under reduced pressure and the residue azeotroped with
toluene three times, then dissolved in tetrahydrofuran (50 mL).
1,8-Diazabicyclo[5.4.0]undec-7-ene (1.84 mL, 12.3 mmol) was added
to the stirred solution under nitrogen at 0.degree. C., followed by
ethyl 3-chloro-3-oxopropionate (1.57 mL, 12.3 mmol) dropwise. The
mixture was stirred at 0.degree. C. for 5 min, and at room
temperature for 3 h, then 1M aqueous hydrochloric acid (50 mL)
added. After stirring a further 0.5 h, water (100 mL) was added and
the mixture extracted with ethyl acetate. The extracts were washed
with brine, dried (MgSO.sub.4), evaporated under reduced pressure
and the residue chromatographed (silica gel, 1-5%
methanol/dichloromethane) to give the title compound (1.60 g) as an
oil. 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 1.18 (t, J=7.08 Hz,
3H) 3.56 (s, 2H) 4.08 (q, J=7.16 Hz, 2H) 4.58 (s, 2H) 4.65 (s, 2H)
7.25-7.36 (m, 5H).
3b) Ethyl
5-hydroxy-6-(1-methylethyl)-3-oxo-2-(phenylmethyl)-2,3-dihydro--
4-pyridazinecarboxylate. A solution of the compound from example
3(a) (0.120 g, 0.508 mmol), ethyl 3-methyl-2-oxobutanoate (0.066 g,
0.458 mmol) and acetic acid (0.030 g, 0.500 mmol) in
dichloromethane (2 mL) was stirred at 50.degree. C. for 18 h, then
cooled, diluted with ethyl acetate (2 mL) and filtered. Solvent was
removed from the filtrate under reduced pressure and the residue
azeotroped 3 times with toluene, then taken up in tetrahydrofuran
(10 mL).
Diazabicyclo[5.4.0]undec-7-ene (0.200 mL, 1.34 mmol) was added and
the solution refluxed under nitrogen for 1.5 h, then cooled and
poured into 1M aqueous hydrochloric acid (10 mL). The mixture was
extracted with ether and the organic extracts washed with 1 M
aqueous sodium hydroxide. The aqueous extracts were washed with
ether, then acidified with 1M aqueous hydrochloric acid and
extracted with ether. The extracts were dried (MgSO.sub.4),
evaporated under reduced pressure and the residue chromatographed
(silica gel, 1-5% methanol/dichloromethane) to give the title
compound (0.029 g, 20%) as a solid. LCMS (ES.sup.+) m/z 317
(MH.sup.+).
3c)
N-{[5-Hydroxy-6-(1-methylethyl)-3-oxo-2-(phenylmethyl)-2,3-dihydro-4--
pyridazinyl]carbonyl}glycine. A stirred mixture of the compound
from example 3(b) (0.029 g, 0.092 mmol), anhydrous glycine, sodium
salt (0.050 g, 0.515 mmol) and 2-methoxyethanol (2 mL) was heated
under reflux under nitrogen for 2 h, cooled and diluted with water
(20 mL). 1M aqueous hydrochloric acid (0.5 mL) was added slowly and
the mixture stirred for 0.5 h, then the solid filtered, washed with
water and dried to leave the title compound (0.025 g, 78%) as a
solid. 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 1.20 (d, J=7.07
Hz, 6H) 3.19 (sept, J=6.82 Hz, 1H) 4.09 (d, J=5.56 Hz, 2H) 5.25 (s,
2H) 7.27-7.37 (m, 5H) 10.19 (t, J=5.05 Hz, 1 H) 12.95 (br s, 1H),
15.87 (br s, 1H).
Example 4
##STR00015##
N-[(5-Hydroxy-3-oxo-6-phenyl-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
4a) Ethyl
3-{2-[2-(ethyloxy)-2-oxo-1-phenylethylidene]hydrazino}-3-oxopro-
panoate. A mixture of ethyl 3-hydrazino-3-oxopropionate (1.46 g,
10.0 mmol), ethyl benzoylformate (2.14 g, 12.0 mmol), acetic acid
(0.5 mL, 8.3 mmol) and dichloromethane (10 mL) was stirred at room
temperature with excess anhydrous magnesium sulfate for 20 h, then
filtered and the cake washed with ethyl acetate. The filtrate was
evaporated under reduced pressure, azeotroped 3 times with toluene
and chromatographed (silica gel, 10-3-% ethyl acetate/hexane) to
give the title compound (0.773 g, 25%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 1.15 (t, J=7.07 Hz, 3H) 1.30 (t, J=7.07
Hz, 3H) 3.74 (s, 2H) 4.08 (q, J=7.07 Hz, 2H) 4.40 (q, J=7.18 Hz,
2H) 7.42-7.49 (m, 3H) 7.55-7.59 (m, 2H) 11.57 (s, 1H).
4b) Ethyl
5-hydroxy-3-oxo-6-phenyl-2,3-dihydro-4-pyridazinecarboxylate. A 1 M
solution of piperidinium acetate in 5% ethanol/toluene (1.10 mL,
1.10 mmol) was added to a solution of the compound from example
4(a) (0.350 g, 1.14 mmol) in toluene (10 mL) and the mixture
stirred under reflux for 1 h. Diazabicyclo[5.4.0]undec-7-ene (0.30
mL, 2.00 mmol) was added and the reflux continued for 2 h. After
cooling, the mixture was partitioned between 1M aqueous
hydrochloric acid and ethyl acetate. The extracts were dried
(MgSO.sub.4), evaporated under reduced pressure and the residue
chromatographed (silica gel, 2-8% methanol/dichloromethane). The
partially purified product was triturated with ether to give the
title compound (0.115 g, 39%) as a solid. 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 1.30 (t, J=7.20 Hz, 3H) 4.32 (q, J=7.16
Hz, 2H) 7.43-7.49 (m, 3H) 7.64-7.67 (m, 2H) 13.05 (s, 1H).
4c)
N-[(5-Hydroxy-3-oxo-6-phenyl-2,3-dihydro-4-pyridazinyl)carbonyl]glyci-
ne. A stirred mixture of the compound from example 4(b) (0.060 g,
0.251 mmol), anhydrous glycine, sodium salt (0.045 g, 0.462 mmol)
and 2-methoxy-ethanol (3 mL) was heated under reflux under nitrogen
for 2 h, cooled and diluted with water (20 mL). After filtering, 1M
aqueous hydrochloric acid (0.5 mL) was added slowly to the
filtrate, then the solid filtered, washed with water and dried to
leave the title compound (0.025 g, 78%) as a white powder. 1H NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 4.02 (d, J=5.56 Hz, 2H)
7.40-7.47 (m, 3H) 7.74-7.80 (m, 2H) 10.45 (s, 1H) 12.94 (s,
1H).
Example 5
##STR00016##
N-[(2-{[4-(1,1-Dimethylethyl)phenyl]methyl}-5-hydroxy-3-oxo-6-phenyl-2,3-d-
ihydro-4-pyridazinyl)carbonyl]glycine
5a) Ethyl
2-{[4-(1,1-dimethylethyl)phenyl]methyl}-5-hydroxy-3-oxo-6-pheny-
l-2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride (0.020 g of a
60% oil suspension, 0.500 mmol) was added to a stirred solution of
the compound from example 4(b) (0.052 g, 0.200 mmol) in
dimethylformamide (1 mL) under nitrogen. After 15 min stirring at
room temperature, 4-t-butylbenzyl bromide (0.037 mL, 0.200 mmol)
was injected and the mixture stirred for 2 h, then partitioned
between 1M aqueous hydrochloric acid and ethyl acetate. The
extracts were washed with water and brine, then dried (MgSO.sub.4),
evaporated under reduced pressure and the residue chromatographed
(silica gel, 0-5% methanol/dichloromethane) to give the title
compound (0.053 g, 65%) as a gum. 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 1.25 (s, 9H) 1.28 (t, J=7.20 Hz, 3H) 4.29 (q, J=7.07
Hz, 2H) 5.20 (s, 2H) 7.25-7.29 (m, 2H) 7.34-7.41 (m, 2H) 7.44-7.50
(m, 3H) 7.67- 7.72 (m, 2H).
5b)
N-[(2-{[4-(1,1-Dimethylethyl)phenyl]methyl}-5-hydroxy-3-oxo-6-phenyl--
2,3-dihydro-4-pyridazinyl)carbonyl]glycine. A stirred mixture of
the compound from example 5(a) (0.052 g, 0.128 mmol), anhydrous
glycine, sodium salt (0.050 g, 0.515 mmol) and 2-methoxyethanol (3
mL) was heated under reflux under nitrogen for 2 h, cooled and
diluted with water (20 mL). After filtering, 1M aqueous
hydrochloric acid was added slowly to the filtrate until the pH had
dropped to 2, then the solid filtered, washed with water and dried
to leave the title compound (0.047 g, 84%) as a pale pink powder.
1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 1.25 (s, 9H) 4.13 (d,
J=5.56 Hz, 2H) 5.32 (s, 2H) 7.28-7.34 (m, 2H) 7.34-7.40 (m, 2H)
7.48-7.51 (m, 3H) 7.76-7.82 (m, 2H) 10.27 (t, J=5.18 Hz, 1H) 12.99
(s, 1H) 16.32 (s, 1H).
Example 6
##STR00017##
N-{[5-Hydroxy-3-oxo-6-phenyl-2-(2-phenylethyl)-2,3-dihydropyridazin-4-yl]c-
arbonyl}glycine
6a) Ethyl
5-hydroxy-3-oxo-6-phenyl-2-(2-phenylethyl)-2,3-dihydro-4-pyrida-
zinecarboxylate. Sodium hydride (0.040 g of a 60% oil suspension,
1.00 mmol) was added to a stirred solution of ethyl
5-hydroxy-3-oxo-6-phenyl-2,3-dihydro-4-pyridazinecarboxylate
(example 4(b), 0.100 g, 0.384 mmol) in dimethylformamide (1.5 mL)
under nitrogen. After 15 min stirring at room temperature, the
mixture was cooled in an ice bath and (2-iodoethyl)benzene (0.056
mL, 0.384 mmol) injected. The mixture was stirred for 18 h while
warming to room temperature, then poured into 0.1M aqueous
hydrochloric acid (50 mL) and extracted with ethyl acetate. The
extracts were washed with water and brine, then dried (MgSO.sub.4),
evaporated under reduced pressure and the residue chromatographed
(silica gel, 1-5% methanol dichloromethane) to give the title
compound (0.077 g, 55%) as a colourless gum. 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 1.30 (t, J=7.07 Hz, 3H) 3.04 (t, J=7.33
Hz, 2H) 4.30 (t, J=7.32 Hz, 2H) 4.32 (q, J=7.07 Hz, 2H) 7.20-7.26
(m, 3H) 7.28-7.35 (m, 2H) 7.40-7.47 (m, 3H) 7.50-7.56 (m, 2H) 12.45
(br. s., 1 H).
6b)
N-{[5-Hydroxy-3-oxo-6-phenyl-2-(2-phenylethyl)-2,3-dihydropyridazin-4-
-yl]carbonyl}glycine. A stirred mixture of ethyl
5-hydroxy-3-oxo-6-phenyl-2-(2-phenylethyl)-2,3-dihydro-4-pyridazinecarbox-
ylate (0.075 g, 0.206 mmol), anhydrous glycine, sodium salt (0.050
g, 0.515 mmol) and 2-methoxyethanol (3 mL) was heated under reflux
under nitrogen for 2 h, then cooled and diluted with water (30 mL).
After filtering, 1M aqueous hydrochloric acid was added slowly to
the filtrate until the pH had dropped to 2, then the solid
filtered, washed with water and dried to leave the title compound
(0.073 g, 90%) as an off-white powder. 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 3.09 (t, J=7.33 Hz, 2H) 4.12 (d, J=5.31
Hz, 2H) 4.40 (t, J=7.33 Hz, 2H) 7.20-7.27 (m, 3H) 7.28-7.35 (m, 2H)
7.41-7.50 (m, 3H) 7.59-7.66 (m, 2H) 10.35 (t, J=5.31 Hz, 1 H).
Example 7
##STR00018##
N-{[2-(2-Cyclopropylethyl)-5-hydroxy-3-oxo-6-phenyl-2,3-dihydro-4-pyridazi-
nyl]carbonyl}glycine
7a) (2-Bromoethyl)cyclopropane. N-Bromosuccinimide (1.60 g, 8.99
mmol) was added in portions to an ice-cooled, stirred solution of
2-cyclopropylethanol (0.760 g, 8.82 mmol) and triphenylphosphine
(2.36 g, 9.00 mmol) in dichloromethane (10 mL) under nitrogen. The
mixture was stirred for 2 h while warming to room temperature, then
diluted with hexane (90 mL) and filtered through a short silica gel
column. The column was washed with 10% ethyl acetate/hexane and the
combined filtrates evaporated under reduced pressure at room
temperature to give the title compound (0.179 g, 14%) as a
colourless liquid. 1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm
0.10-0.17 (m, 2H) 0.47-0.56 (m, 2H) 0.79-0.90 (m, 1H) 1.79 (q,
J=6.91 Hz, 2H) 3.47 (t, J=7.07 Hz, 2H).
7b) Ethyl
2-(2-cyclopropylethyl)-5-hydroxy-3-oxo-6-phenyl-2,3-dihydro-4-p-
yridazinecarboxylate. Sodium hydride (0.040 g of a 60% oil
suspension, 1.00 mmol) was added to a stirred solution of ethyl
5-hydroxy-3-oxo-6-phenyl-2,3-dihydro-4-pyridazinecarboxylate
(example 4(b), 0.100 g, 0.384 mmol) in dimethylformamide (1.5 mL)
under nitrogen. After 15 min stirring at room temperature, the
mixture was cooled in an ice bath and (2-bromoethyl)cyclopropane
(0.057 g, 0.384 mmol) added. The mixture was stirred for 18 h while
warming to room temperature, then poured into 0.1M aqueous
hydrochloric acid (50 mL) and extracted with ethyl acetate. The
extracts were washed with water and brine, then dried (MgSO.sub.4),
evaporated under reduced pressure and the residue chromatographed
(silica gel, 1-5% methanol/dichloromethane) to give the title
compound (0.067 g, 53%) as a colourless gum. 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm -0.05-0.06 (m, 2H) 0.34-0.45 (m, 2H)
0.64-0.78 (m, 1H) 1.29 (t, J=7.07 Hz, 3H) 1.62 (q, J=7.07 Hz, 2H)
4.14 (t, J=7.20 Hz, 2H) 4.31 (q, J=7.07 Hz, 2H) 7.41-7.52 (m, 3H)
7.63-7.71 (m, 2H) 12.42 (br. s., 1H).
7c)
N-{[2-(2-Cyclopropylethyl)-5-hydroxy-3-oxo-6-phenyl-2,3-dihydro-4-pyr-
idazinyl]carbonyl}glycine. A stirred mixture of ethyl
2-(2-cyclopropylethyl)-5-hydroxy-3-oxo-6-phenyl-2,3-dihydro-4-pyridazinec-
arboxylate (0.065 g, 0.198 mmol), anhydrous glycine, sodium salt
(0.050 g, 0.515 mmol) and 2-methoxyethanol (3 mL) was heated under
reflux under nitrogen for 2 h, then cooled and diluted with water
(30 mL). After filtering, 1M aqueous hydrochloric acid was added
slowly to the filtrate until the pH had dropped to 2, then the
solid filtered, washed with water and dried to leave the title
compound (0.062 g, 87%) as an off-white powder. 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm -0.05-0.09 (m, 2H) 0.35-0.47 (m, 2H)
0.66-0.79 (m, 1H) 1.68 (q, J=7.07 Hz, 2H) 4.15 (d, J=5.56 Hz, 2H)
4.26 (t, J=7.07 Hz, 2H) 7.41-7.56 (m, 3H) 7.69-7.84 (m, 2H) 10.35
(br t, J=5.05 Hz, 1H) 13.00 (br. s., 1H) 16.25 (s, 1H).
Example 8
##STR00019##
N-{[2-(2-Chlorobenzyl)-5-hydroxy-3-oxo-6-phenyl-2,3-dihydropyridazin-4-yl]-
carbonyl}glycine
8a) Ethyl
2-[(2-chlorophenyl)methyl]-5-hydroxy-3-oxo-6-phenyl-2,3-dihydro-
-4-pyridazinecarboxylate. Sodium hydride (0.040 g of a 60% oil
suspension, 1.00 mmol) was added to a stirred solution of ethyl
5-hydroxy-3-oxo-6-phenyl-2,3-dihydro-4-pyridazinecarboxylate
(example 4(b), 0.100 g, 0.384 mmol) in dimethylformamide (1.5 mL)
under nitrogen. After 15 min stirring at room temperature, the
mixture was cooled in an ice bath and 2-chlorobenzyl bromide (0.051
mL, 0.384 mmol) injected. The mixture was stirred for 18 h while
warming to room temperature, then poured into 0.1M aqueous
hydrochloric acid (50 mL) and extracted with ethyl acetate. The
extracts were washed with water and brine, then dried (MgSO.sub.4),
evaporated under reduced pressure and the residue chromatographed
(silica gel, 1-5% methanol/dichloromethane) to give the title
compound (0.098 g, 66%) as a colourless gum. 1H NMR (400 MHz,
CHLOROFORM-d) .delta. ppm 1.50 (t, J=7.20 Hz, 3H) 4.55 (q, J=7.07
Hz, 2H) 5.54 (s, 2 H) 7.20-7.28 (m, 3H) 7.35-7.52 (m, 4H) 7.75-7.84
(m, 2H) 13.83 (s, 1H).
8b)
N-{[2-(2-Chlorobenzyl)-5-hydroxy-3-oxo-6-phenyl-2,3-dihydropyridazin--
4-yl]carbonyl}glycine. A stirred mixture of ethyl
2-[(2-chlorophenyl)methyl]-5-hydroxy-3-oxo-6-phenyl-2,3-dihydro-4-pyridaz-
inecarboxylate (0.098 g, 0.255 mmol), anhydrous glycine, sodium
salt (0.050 g, 0.515 mmol) and 2-methoxyethanol (3 mL) was heated
under reflux under nitrogen for 2 h, then cooled and diluted with
water (30 mL). After filtering, 1M aqueous hydrochloric acid was
added slowly to the filtrate until the pH had dropped to 2, then
the solid filtered, washed with water and dried to leave the title
compound (0.099 g, 93%) as an off-white powder. 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 4.14 (d, J=5.81 Hz, 2H) 5.46 (s, 2H)
7.23-7.26 (m, 1H) 7.29-7.40 (m, 2H) 7.44-7.49 (m, 3H) 7.52 (dd,
J=7.45, 1.64 Hz, 1H) 7.71-7.79 (m, 2H) 10.21 (br t, J=4.80 Hz, 1H)
12.98 (br. s., 1H) 16.41 (br. s., 1H).
Example 9
##STR00020##
N-{[5-Hydroxy-3-oxo-2-(phenylmethyl)-6-(trifluoromethyl)-2,3-dihydro-4-pyr-
idazinyl]carbonyl}glycine
9a) Ethyl
5-hydroxy-3-oxo-6-(trifluoromethyl)-2,3-dihydro-4-pyridazinecar-
boxylate. A mixture of ethyl 3-hydrazino-3-oxopropionate (0.438 g,
3.00 mmol), ethyl 3,3,3-trifluoro-2-oxopropionate (0.510 g, 3.00
mmol), p-toluenesulfonic acid monohydrate acid (0.057 g, 0.300
mmol) and toluene (10 mL) was refluxed using a Dean and Stark trap
to remove water for 3 h, then cooled. Magnesium sulfate was added
and the mixture filtered through a short silica gel column. The
column was washed with 50% ethyl acetate/hexane and the combined
filtrates evaporated under reduced pressure. The residue was
chromatographed (silica gel, 10-40% ethyl acetate/hexane) to give
the intermediate hydrazone. Diazabicyclo[5.4.0]undec-7-ene (0.083
mL, 0.558 mmol) was added to a solution of the hydrazone in dioxane
(5 mL) and the mixture refluxed under nitrogen for 1 h. After
cooling, 0.1M aqueous hydrochloric acid (50 mL) was added and the
mixture extracted with ethyl acetate. The extracts were dried
(MgSO.sub.4) and evaporated under reduced pressure to give the
title compound (0.085 g, 12%) as a light brown powder. 1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 1.26 (t, J=7.07 Hz, 3H) 4.25 (q,
J=7.07 Hz, 2H) 13.30 (s, 1H).
9b) Ethyl
5-hydroxy-3-oxo-2-(phenylmethyl)-6-(trifluoromethyl)-2,3-dihydr-
o-4-pyridazinecarboxylate. Sodium hydride (0.040 g of a 60% oil
suspension, 1.00 mmol) was added to a stirred solution of ethyl
5-hydroxy-3-oxo-6-(trifluoromethyl)-2,3-dihydro-4-pyridazinecarboxylate
(0.083 g, 0.329 mmol) in dimethylformamide (1.5 mL) under nitrogen.
After 10 min stirring at room temperature, the mixture was cooled
in an ice bath and benzyl bromide (0.043 mL, 0.362 mmol) injected.
The mixture was stirred for 3 h at 0.degree. C. 0.1M aqueous
hydrochloric acid (50 mL) was added and the mixture extracted with
ethyl acetate. The extracts were washed with water and brine, then
dried (MgSO.sub.4), evaporated under reduced pressure and the
residue chromatographed (silica gel, 5-10% methanol
dichloromethane) to give the title compound (0.050 g, 44%). 1H NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 1.23 (t, J=6.95 Hz, 3H) 4.17
(q, J=7.07 Hz, 2H) 5.14 (s, 2H) 7.12-7.44 (m, 5H).
9c)
N-{[5-Hydroxy-3-oxo-2-(phenylmethyl)-6-(trifluoromethyl)-2,3-dihydro--
4-pyridazinyl]carbonyl}glycine. A stirred mixture of ethyl
5-hydroxy-3-oxo-2-(phenylmethyl)-6-(trifluoromethyl)-2,3-dihydro-4-pyrida-
zinecarboxylate (0.048 g, 0.140 mmol), anhydrous glycine, sodium
salt (0.050 g, 0.515 mmol) and 2-methoxyethanol (3 mL) was heated
under reflux under nitrogen for 2 h, then cooled and diluted with
water (30 mL). After filtering, 1M aqueous hydrochloric acid was
added slowly to the filtrate until the pH had dropped to 2. The
mixture was extracted with ethyl acetate and the extracts dried
(MgSO.sub.4), then evaporated under reduced pressure. The residue
was purified by HPLC (ODS, 10-90% acetonitrile/water+0.1%
trifluoroacetic acid). The product was reprecipitated from 1M
aqueous sodium hydroxide with 1M aqueous hydrochloric acid and the
solid filtered, washed with water and dried to give the title
compound (0.014 g, 27%) as a white solid. 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 4.12 (d, J=5.81 Hz, 2H) 5.34 (s, 2H)
7.27-7.41 (m, 5H) 9.96 (t, J=5.43 Hz, 1H) 13.02 (br. s., 1H).
Example 10
##STR00021##
N-{[6-Cyclohexyl-5-hydroxy-3-oxo-2-(phenylmethyl)-2,3-dihydro-4-pyridaziny-
l]carbonyl}glycine
10a) Ethyl
6-cyclohexyl-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylate. A
mixture of ethyl 3-hydrazino-3-oxopropionate (0.450 g, 3.08 mmol),
ethyl cyclohexyl(oxo) acetate (0.600 g, 3.26 mmol),
p-toluenesulfonic acid monohydrate acid (0.060 g, 0.315 mmol) and
toluene (15 mL) was refluxed using a Dean and Stark trap to remove
water for 1 h, then cooled and chromatographed (silica gel, 10-40%
ethyl acetate/hexane) to give the intermediate hydrazone.
Diazabicyclo[5.4.0]undec-7-ene (0.540 mL, 3.61 mmol) was added to a
solution of the hydrazone, pre-dried by azeotroping with toluene,
in dioxane (10 mL) and the mixture refluxed under nitrogen for 19
h. After cooling, 0.1M aqueous hydrochloric acid (50 mL) was added
and the mixture extracted with ethyl acetate. The extracts were
washed with brine, dried (MgSO.sub.4) and evaporated under reduced
pressure. The residue was chromatographed (silica gel, 1-5%
methanol/dichloromethane) to give the title compound (0.404 g, 52%)
as a cream solid. 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
1.12-1.23 (m, 1H) 1.27 (t, J=7.07 Hz, 3H) 1.29-1.38 (m, 4H)
1.64-1.71 (m, 1H) 1.72-1.90 (m, 4H) 2.74-2.86 (m, 1H) 4.28 (q,
J=7.16 Hz, 2H) 12.30 (br. s., 1H) 12.62 (s, 1H).
10b) Ethyl
6-cyclohexyl-5-hydroxy-3-oxo-2-(phenylmethyl)-2,3-dihydro-4-pyridazinecar-
boxylate. Sodium hydride (0.040 g of a 60% oil suspension, 1.00
mmol) was added to a stirred suspension of ethyl
6-cyclohexyl-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylate
(0.100 g, 0.376 mmol) in dimethylformamide (1.5 mL) under nitrogen.
After 10 min stirring at room temperature, the mixture was cooled
in an ice bath and benzyl bromide (0.045 mL, 0.378 mmol) injected.
The mixture was stirred for 0.5 h at 0.degree. C. and 0.5 h at room
temperature. 0.1M aqueous hydrochloric acid (50 mL) was added and
the mixture extracted with ethyl acetate. The extracts were washed
with water and brine, then dried (MgSO.sub.4), evaporated under
reduced pressure and the residue chromatographed (silica gel, 0-4%
methanol/dichloromethane) to give the title compound (0.095 g, 71%)
as a colourless oil. LCMS (ES.sup.+) m/z 357 (MH.sup.+).
10c)
N-{[6-Cyclohexyl-5-hydroxy-3-oxo-2-(phenylmethyl)-2,3-dihydro-4-pyri-
dazinyl]carbonyl}glycine. A stirred mixture of ethyl
6-cyclohexyl-5-hydroxy-3-oxo-2-(phenylmethyl)-2,3-dihydro-4-pyridazinecar-
boxylate (0.095 g, 0.267 mmol), anhydrous glycine, sodium salt
(0.053 g, 0.544 mmol) and 2-methoxyethanol (3 mL) was heated under
reflux under nitrogen for 2 h, then cooled and diluted with water
(30 mL). 1M aqueous hydrochloric acid was added slowly until the pH
had dropped to 2. After 60 h, the solid was filtered, washed with
water and dried to leave the title compound (0.095 g, 92%) as a
white solid. 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 1.12-1.28
(m, 1H) 1.28-1.49 (m, 4H) 1.63-1.72 (m, 1H) 1.74-1.81 (m, 2 H)
1.83-1.91 (m, 2H) 2.82-2.94 (m, 1H) 4.09 (d, J=5.56 Hz, 2H) 5.25
(s, 2H) 7.25-7.31 (m, 3 H) 7.31-7.38 (m, 2H) 10.19 (t, J=5.18 Hz,
1H) 12.97 (br. s., 1H) 15.86 (br. s., 1H).
Example 11
##STR00022##
N-[(6-Cyclohexyl-2-{[4-(1,1-dimethylethyl)phenyl]methyl}-5-hydroxy-3-oxo-2-
,3-dihydro-4-pyridazinyl)carbonyl]glycine
11a) Ethyl
6-cyclohexyl-2-{[4-(1,1-dimethylethyl)phenyl]methyl}-5-hydroxy-3-oxo-2,3--
dihydro-4-pyridazinecarboxylate. Sodium hydride (0.040 g of a 60%
oil suspension, 1.00 mmol) was added to a stirred suspension of
ethyl
6-cyclohexyl-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylate
(example 10(a), 0.100 g, 0.376 mmol) in dimethylformamide (2.0 mL)
under nitrogen. After 15 min stirring at room temperature, the
mixture was cooled in an ice bath and 4-tert-butylbenzyl bromide
(0.071 mL, 0.384 mmol) injected. The mixture was stirred for 0.5 h
at 0.degree. C. and 3 h at room temperature. 0.1M aqueous
hydrochloric acid (50 mL) was added and the mixture extracted with
ethyl acetate. The extracts were washed with dilute aqueous sodium
chloride, then dried (MgSO.sub.4), evaporated under reduced
pressure and the residue chromatographed (silica gel, 0-6%
methanol/dichloromethane) to give the title compound (0.087 g, 56%)
as a gum. LCMS (ES.sup.+) m/z 413 (MH.sup.+).
11b)
N-[(6-Cyclohexyl-2-{[4-(1,1-dimethylethyl)phenyl]methyl}-5-hydroxy-3-
-oxo-2,3-dihydro-4-pyridazinyl)carbonyl]glycine. A stirred mixture
of ethyl
6-cyclohexyl-2-{[4-(1,1-dimethylethyl)phenyl]methyl}-5-hydroxy-3-ox-
o-2,3-dihydro-4-pyridazinecarboxylate (0.087 g, 0.211 mmol),
anhydrous glycine, sodium salt (0.050 g, 0.515 mmol) and
2-methoxyethanol (3 mL) was heated under reflux under nitrogen for
2 h, then cooled and diluted with water (30 mL). The mixture was
filtered and 1M aqueous hydrochloric acid added slowly until the pH
had dropped to 2. After 18 h, the solid was filtered, washed with
water, dried and chromatographed (silica gel, 10%
methanol/dichloromethane). The product was precipitated from
methanol with water, and the solid collected, washed with water,
then dried to leave the title compound (0.057 g, 61%) as a solid.
1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 1.18-1.24 (m, 1H) 1.25
(s, 9H) 1.29-1.50 (m, 4 H) 1.64-1.73 (m, 1H) 1.76-1.81 (m, 2H)
1.83-1.92 (m, 2H) 2.79-2.97 (m, 1H) 4.09 (d, J=5.81 Hz, 2H) 5.20
(s, 2H) 7.23 (d, J=8.34 Hz, 2H) 7.36 (d, J=8.34 Hz, 2H) 10.19 (t,
J=5.56 Hz, 1H) 12.94 (br. s., 1H) 15.83 (s, 1H).
Example 12
##STR00023##
N-({2-[(2-Chlorophenyl)methyl]-6-cyclohexyl-5-hydroxy-3-oxo-2,3-dihydro-4--
pyridazinyl}carbonyl)glycine
12a) Ethyl
2-[(2-chlorophenyl)methyl]-6-cyclohexyl-5-hydroxy-3-oxo-2,3-dihydro-4-pyr-
idazinecarboxylate. Sodium hydride (0.040 g of a 60% oil
suspension, 1.00 mmol) was added to a stirred suspension of ethyl
6-cyclohexyl-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylate
(example 10(a), 0.100 g, 0.376 mmol) in dimethylformamide (2.0 mL)
under nitrogen. After 15 min stirring at room temperature, the
mixture was cooled in an ice bath and 2-chlorobenzyl bromide (0.051
mL, 0.384 mmol) injected. The mixture was stirred for 0.5 h at
0.degree. C. and 3 h at room temperature. 0.1M aqueous hydrochloric
acid (50 mL) was added and the mixture extracted with ethyl
acetate. The extracts were washed with dilute aqueous sodium
chloride, then dried (MgSO.sub.4), evaporated under reduced
pressure and the residue chromatographed (silica gel, 0-6%
methanol/dichloromethane) to give the title compound (0.093 g, 63%)
as a gum. LCMS (ES.sup.+) m/z 391 (MH.sup.+).
12b)
N-({2-[(2-Chlorophenyl)methyl]-6-cyclohexyl-5-hydroxy-3-oxo-2,3-dihy-
dro-4-pyridazinyl}carbonyl)glycine. A stirred mixture of ethyl
2-[(2-chlorophenyl)methyl]-6-cyclohexyl-5-hydroxy-3-oxo-2,3-dihydro-4-pyr-
idazinecarboxylate (0.093 g, 0.238 mmol), anhydrous glycine, sodium
salt (0.050 g, 0.515 mmol) and 2-methoxyethanol (3 mL) was heated
under reflux under nitrogen for 2 h, then cooled and diluted with
water (30 mL). The mixture was filtered and 1M aqueous hydrochloric
acid added slowly until the pH had dropped to 2. After 18 h, the
solid was filtered, washed with water, dried and chromatographed
(silica gel, 10% methanol/dichloromethane). The product was
precipitated from methanol with water, and the solid collected,
washed with water, then dried to leave the title compound (0.041 g,
41%) as a solid. LCMS (ES.sup.+) m/z 420 (MH.sup.+).
Example 13
##STR00024##
N-[(2,6-Dicyclohexyl-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinyl)carbonyl]gl-
ycine
13a) Ethyl
2,6-dicyclohexyl-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylate.
A mixture of cyclohexylhydrazine hydrochloride (0.451 g, 3.00
mmol), ethyl cyclohexyl (oxo)acetate (0.500 g, 2.71 mmol),
anhydrous sodium acetate (0.246 g, 0.300 mmol) and dichloromethane
(5 mL) was stirred at room temperature for 18 h, then diluted with
water and extracted with dichloromethane. The extracts were dried
(MgSO.sub.4), evaporated under reduced pressure and the residue
chromatographed (silica gel, 5-30% ethyl acetate/hexane) to give
the intermediate hydrazone. Diazabicyclo[5.4.0]undec-7-ene (0.260
mL, 1.74 mmol) was added to a solution of the hydrazone in
tetrahydrofuran (10 mL) under nitrogen. The mixture was cooled in
ice and ethyl 3-chloro-3-oxopropionate (0.223 mL, 1.74 mmol)
injected dropwise. The mixture was stirred at room temperature for
2 h, then more diazabicyclo[5.4.0]undec-7-ene (0.520 mL, 3.48 mmol)
added and the mixture refluxed under nitrogen for 2 h. After
cooling, 0.1M aqueous hydrochloric acid (100 mL) was added and the
mixture extracted with ethyl acetate. The extracts were dried
(MgSO.sub.4), evaporated under reduced pressure and the residue
chromatographed (silica gel, 2-5%, then 50% ethyl acetate/hexane)
to give the title compound (0.184 g, 19%) as a white solid. 1H NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 1.13-1.21 (m, 2H) 1.26 (t,
J=7.07 Hz, 3H) 1.31-1.45 (m, 6H) 1.53-1.91 (m, 12H) 2.74-2.90 (m,
1H) 4.26 (q, J=7.07 Hz, 2H) 4.58-4.72 (m, 1 H) 12.02 (br. s., 1
H).
13b)
N-[(2,6-Dicyclohexyl-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinyl)carbo-
nyl]glycine. A stirred mixture of ethyl
2,6-dicyclohexyl-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylate
(0.182 g, 0.522 mmol), anhydrous glycine, sodium salt (0.102 g,
1.05 mmol) and 2-methoxyethanol (5 mL) was heated under reflux
under nitrogen for 2 h, then cooled and diluted with water (50 mL).
1M aqueous hydrochloric acid was added slowly until the pH had
dropped to 2. The solid was filtered, washed with water and dried
to leave the title compound (0.156 g. 79%) as a cream powder. 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 1.12-1.28 (m, 2H) 1.29-1.49
(m, 6H) 1.55-1.97 (m, 12H) 2.80-2.91 (m, 1H) 4.09 (d, J=5.56 Hz,
2H) 4.64-4.85 (m, 1H) 10.35 (t, J=5.31 Hz, 1H) 15.76 (br. s.,
1H).
Example 14
##STR00025##
N-{[2-{[4-(1,1-Dimethylethyl)phenyl]methyl}-5-hydroxy-6-(1-methylethyl)-3--
oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
14a) Ethyl
5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinecarboxylate.
In 3 separate microwave tubes was added
Ethyl-3-methyl-2-oxobutyrate (5 g, 34.7 mmol) and Ethyl-malonyl
hydrazide (6.08 g, 41.6 mmol) in Ethanol (10 ml) and Acetic Acid
(0.5 ml). The reactions were irridatiated at 150.degree. C. for 20
minutes. The crude reaction mixture was evaporated down to give a
yellow oil. The 3 crude oils were separately resuspended in
1,4-Dioxane (12 ml) and DBU (7.84 ml, 52.0 mmol) was added. The
solution was divided into 2 microwave tubes and the reactions were
irridatiated at 150.degree. C. for 20 minutes. The fractions were
combined, diluted with water (80 ml) and acidified slowly (over 15
minutes) with 6N HCl to cause a precipitate. The precipitate was
collected by filtration and dried to give the product as an off
white solid (11.12 g, 46.7 mmol, 44.9% yield). 1H NMR (400 MHz,
DMSO-d.sub.6) d ppm 12.64 (s, 1H), 12.31 (s, 1 H), 4.28 (q, J=7.16
Hz, 2H), 3.13 (sept, J=6.82 Hz, 1H), 1.27 (t, J=7.07 Hz, 3 H), 1.14
(d, J=6.82 Hz, 6H). MS (ES+) m/e 227 [M+H]+.
14b) Ethyl
2-{[4-(1,1-dimethylethyl)phenyl]methyl}-5-hydroxy-6-(1-methylethyl)-3-oxo-
-2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride (49 mg, 1.22
mmol) was added to a solution of the compound from example 14a)
(110 mg, 0.49 mmol) in N,N-Dimethylformamide (DMF) at 0.degree. C.
The reaction was brought to room temperature and stirred for 40
minutes. The temperature was then reduced to 0.degree. C. and
4-tert-butylbenzyl bromide (0.09 mL, 0.49 mmol) was added. The
reaction was brought to room temperature and stirred for 3 hours.
The solution was diluted with EtOAc and H.sub.2O and the layers
separated. 1N HCl was added to the aqueous layer and it was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 25-40% EtOAc/Hexanes) to give the title compound (111
mg, 61%). 1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 13.29 (s, 1H)
7.31-7.43 (m, 4H) 5.24 (s, 2H) 4.49 (q, J=7.07 Hz, 2 H) 3.25 (sept,
J=6.82 Hz, 1H) 1.46 (t, J=7.20 Hz, 3H) 1.31 (s, 9H) 1.27 (d, J=6.82
Hz, 6H). 14c)
N-{[2-{[4-(1,1-Dimethylethyl)phenyl]methyl}-5-hydroxy-6-(1-methylethyl)-3-
-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium
salt (38 mg, 0.39 mmol) was added to a solution of the compound
from example 14b) (73 mg, 0.20 mmol) in 2-methoxyethanol at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes to give the title compound (80 mg, 29%). 1H
NMR (400 MHz, CHLOROFORM-d) .delta. ppm 15.08 (s, 1H) 10.50 (t,
J=6.06 Hz, 1H) 7.36 (d, J=1.52 Hz, 4H) 5.27 (s, 2H) 4.24 (d, J=5.81
Hz, 2H) 3.27 (sept, J=6.86 Hz, 1H) 1.31 (s, 9H) 1.28 (d, J=6.82 Hz,
6H).
Example 15
##STR00026##
N-{[2-[(2-Chlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine
15a) Ethyl
2-[(2-chlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro--
4-pyridazinecarboxylate. Sodium hydride (37 mg, 0.93 mmol) was
added to a solution of the compound from example 14a) (84 mg, 0.37
mmol) in N,N-Dimethylformamide (DMF) (1.5 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 40
minutes. The temperature was then reduced to 0.degree. C. and
2-chlorobenzyl bromide (48 .mu.L, 0.37 mmol) was added. The
reaction was brought to room temperature and stirred for 3 hours
followed by the addition of 1N HCl. The solution was diluted with
EtOAc and H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 30-45% EtOAc/Hexanes) to give the title compound (71
mg, 55%). 1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 13.38 (s, 1H)
7.35-7.42 (m, 1H) 7.13-7.26 (m, 3 H) 5.42 (s, 2H) 4.50 (q, J=7.07
Hz, 2H) 3.24 (sept, J=6.82 Hz, 1H) 1.47 (t, J=7.07 Hz, 3H) 1.21 (d,
J=6.82 Hz, 6H).
15b)
N-{[2-[(2-Chlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-
-dihydro-4-pyridazinyl]carbonyl}. Glycine, sodium salt (35 mg, 0.36
mmol) was added to a solution of the compound from example 15a) (64
mg, 0.18 mmol) in 2-methoxyethanol at room temperature. The
reaction was heated to reflux and stirred for 2 h. The reaction was
cooled back to room temperature and H.sub.2O was added. The
solution was filtered and 1N HCl was added to precipitate the
product. The solid was filtered and washed with H.sub.2O and
Hexanes. The product was purified by precipitation from
CH.sub.2Cl.sub.2/Hexanes to give the title compound (39 mg, 57%).
1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 15.22 (s, 1H) 10.42 (t,
J=5.43 Hz, 1H) 7.38-7.45 (m, 1H) 7.19-7.27 (m, 2H) 7.08-7.13 (m,
1H) 5.45 (s, 2H) 4.25 (d, J=5.81 Hz, 2H) 3.27 (sept, J=6.82 Hz, 1
H) 1.22 (d, J=6.82 Hz, 6H).
Example 16
##STR00027##
N-{[2-[(3,5-Difluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-d-
ihydro-4-pyridazinyl]carbonyl}glycine
16a)
Ethyl-2-[(3,5-difluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-o-
xo-2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride (32 mg, 0.80
mmol) was added to a solution of the compound from example 14a) (72
mg, 0.32 mmol) in N,N-Dimethylformamide (DMF) (1.5 mL) at 0.degree.
C. The reaction was brought to room temperature and stirred for 40
minutes. The temperature was then reduced to 0.degree. C. and
3,5-difluorobenzyl bromide (0.04 mL, 0.32 mmol) was added. The
reaction was brought to room temperature and stirred overnight.
Little conversion to product was observed so the reaction was
cooled back to 0.degree. C. and additional NaH (10 mg, 0.25) and
3,5-difluorobenzyl bromide (0.01 mL, 0.08 mmol) were added. After
several hours, the reaction was quenched with 1N HCl. The solution
was diluted with EtOAc and H.sub.2O and the layers separated. The
aqueous layer was backextracted with EtOAc several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The product was purified by column
chromatography (SiO.sub.2, 30-45% EtOAc/Hexanes) to give the title
compound (34 mg, 30%). 1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm
13.40 (s, 1H) 6.94 (ddd, J=14.46, 6.63, 2.15 Hz, 2H) 6.73 (tt,
J=8.97, 2.27 Hz, 1H) 5.22 (s, 2H) 4.51 (q, J=7.16 Hz, 2H) 3.27
(sept, J=6.86 Hz, 1H) 1.47 (t, J=7.20 Hz, 3H) 1.27 (d, J=6.82 Hz,
6H).
16b)
N-{[2-[(3,5-Difluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-
-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt
(18 mg, 0.18 mmol) was added to a solution of the compound from
example 16a) (32 mg, 0.09 mmol) in 2-methoxyethanol (0.9 mL) at
room temperature. The reaction was heated to reflux and stirred for
2 h. The reaction was cooled back to room temperature and H.sub.2O
was added. The solution was filtered and 1N HCl was added to
precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes. The solid was redissolved in hot MeOH and
filtered. The solvent was removed under reduced pressure to give
the title compound as a white solid (26 mg, 75%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 15.92 (s, 1H) 12.96 (s, 1H) 10.13 (t,
J=5.31 Hz, 1H) 7.17 (m, 1H) 6.95-7.05 (m, 2H) 5.27 (s, 2H) 4.09 (d,
J=5.56 Hz, 2H) 3.19 (sept, J=6.78 Hz, 1H) 1.19 (d, J=6.82 Hz,
6H).
Example 17
##STR00028##
N-{[2-(4-Biphenylylmethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-
-pyridazinyl]carbonyl}glycine
17a)
Ethyl-(4-biphenylylmethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dih-
ydro-4-pyridazinecarboxylate. Sodium hydride (40 mg, 0.99 mmol) was
added to a solution of the compound from example 14a) (90 mg, 0.40
mmol) in N,N-Dimethylformamide (DMF) (2.2 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 45
minutes. The temperature was then reduced to 0.degree. C. and
4-(bromomethyl)-biphenyl (98 mg, 0.40 mmol) was added. The reaction
was brought to room temperature and stirred for 3 h. The reaction
was quenched with 1N HCl. The solution was diluted with EtOAc and
H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 25-45% EtOAc/Hexanes) to give the title compound (130
mg, 83%). 1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 13.33 (s, 1H)
7.50-7.61 (m, 6H) 7.41-7.48 (m, 2 H) 7.35 (tt, J=7.33, 1.26 Hz, 1H)
5.31 (s, 2H) 4.50 (q, J=7.07 Hz, 2H) 3.27 (sept, J=6.82 Hz, 1 H)
1.46 (t, J=7.07 Hz, 3H) 1.28 (d, J=6.82 Hz, 6H).
17b)
N-{[2-(4-biphenylylmethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dih-
ydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt (18 mg,
0.18 mmol) was added to a solution of the compound from example
17a) (32 mg, 0.09 mmol) in 2-methoxyethanol (0.9 mL) at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes. The solid was redissolved in hot MeOH and
filtered. The solvent was removed under reduced pressure to give
the title compound as a white solid (26 mg, 75%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 15.89 (s, 1H) 12.98 (s, 1H) 10.20 (t,
J=4.93 Hz, 1H) 7.64 (d, J=7.83 Hz, 4H) 7.30-7.51 (m, 5H) 5.30 (s,
2H) 4.10 (d, J=5.56 Hz, 2H) 3.13-3.27 (m, 1H) 1.22 (d, J=7.07 Hz, 6
H).
Example 18
##STR00029##
N-{[2-(2-Cyclohexylethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4--
pyridazinyl]carbonyl}
18a) Ethyl
2-(2-cyclohexylethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyr-
idazinecarboxylate. Sodium hydride (40 mg, 0.99 mmol) was added to
a solution of the compound from example 14a) (90 mg, 0.40 mmol) in
N,N-Dimethylformamide (DMF) (2.2 mL) at 0.degree. C. The reaction
was brought to room temperature and stirred for 45 minutes. The
temperature was then reduced to 0.degree. C. and 2-cyclohexyl ethyl
bromide (0.09 mL, 0.40 mmol) was added. The reaction was brought to
room temperature and stirred for 4 hours followed by the addition
of 1N HCl. The solution was diluted with EtOAc and H.sub.2O and the
layers separated. The aqueous layer was backextracted with EtOAc
several times. The combined organic layers were washed with Brine,
dried (MgSO.sub.4), filtered and concentrated. The product was
purified by column chromatography (SiO.sub.2, 25-45% EtOAc/Hexanes)
to give the title compound (82 mg, 61%). LCMS (ES.sup.+) m/z 337.0
(MH.sup.+).
18b)
N-{[2-(2-Cyclohexylethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihy-
dro-4-pyridazinyl]carbonyl}. Glycine, sodium salt (47 mg, 0.49
mmol) was added to a solution of the compound from example 18a) (82
mg, 0.24 mmol) in 2-methoxyethanol (1.2 mL) at room temperature.
The reaction was heated to reflux and stirred for 2 h. The reaction
was cooled back to room temperature and H.sub.2O was added. The
solution was filtered and 1N HCl was added to precipitate the
product. The solid was filtered and washed with H.sub.2O and
Hexanes. The product was purified by precipitation from
Et.sub.2O/Hexanes to give the title compound as a white solid (40
mg, 45%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 15.77 (s, 1H)
12.98 (s, 1H) 10.28 (t, J=5.31 Hz, 1H) 4.04-4.16 (m, 4H) 3.18
(sept, J=6.78 Hz, 1H) 1.47-1.82 (m, 7H) 1.20-1.29 (m, 1H) 1.19 (d,
J=6.82 Hz, 6H) 1.06-1.17 (m, 3H) 0.82-0.99 (m, 2H).
Example 19
##STR00030##
N-{[2-(4-Biphenylylmethyl)-6-(4-fluorophenyl)-5-hydroxy-3-oxo-2,3-dihydro--
4-pyridazinyl]carbonyl}glycine
19a) Ethyl (4-fluorophenyl)(oxo)acetate. A solution of diethyl
oxylate (1.4 mL, 10.3 mmol) in THF (40 mL) and Et.sub.2O (40 mmol)
was cooled to -78.degree. C. 4-Fluorophenyl magnesium bromide (2.0M
solution in Et.sub.2O, 6.2 mL, 12.4.0 mmol) was dropwise added and
the solution stirred under a nitrogen atmosphere for 1.5 h at
-78.degree. C. The reaction was brought to 0.degree. C. and
quenched with 6N HCl. Additional Et.sub.2O and H.sub.2O were added
and the layers separated. The aqueous phase was backextracted with
Et.sub.2O several times. The combined organic layers were washed
with Brine, dried (MgSO.sub.4), filtered and concentrated. The
product was purified by column chromatography (SiO.sub.2, 5-10%
EtOAc/Hexanes) to give the title compound as a pale yellow oil
(1.74 g, 86%). 1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 8.05-8.15
(m, 2H) 7.16-7.25 (m, 2H) 4.47 (q, J=7.16 Hz, 2H) 1.45 (t, J=7.07
Hz, 3H).
19b) Ethyl
3-{(2Z)-2-[2-(ethyloxy)-1-(4-fluorophenyl)-2-oxoethylidene]hydrazino}-3-o-
xopropanoate. Ethyl-3-hydrazino-3-oxopropionate (1.31 g, 8.99 mmol)
and catalytic AcOH (0.09 mL, 1.50 mmol) were added to a solution of
the compound from example 19a) in EtOH (15 mL). The reaction was
heated to reflux and stirred overnight. A few spatula tips of MgSO4
were added and the reaction continued to stir for 3 h. The reaction
was then cooled to room temperature and filtered. The filtrate was
concentrated and azeotroped with Toluene several times. The product
was purified by column chromatography (SiO.sub.2, 15-45%
EtOAc/Hexanes) to give the title compound (1.77 g, 73%). 1H NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 10.63-11.68 (m, 1H) 7.22-7.74
(m, 4H) 4.00-4.49 (m, 4H) 3.40-3.80 (m, 2H) 1.04-1.38 (m, 6H). 19c)
Ethyl
6-(4-fluorophenyl)-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylate.
KHMDS (1.60 g, 8.02 mmol) was added in several portions to a
solution of the compound from example 19b) (1.75 g, 5.35 mmol) in
1,4-dioxane (13 mL) at room temperature. The reaction was heated to
reflux and stirred for 2.5 h. The reaction was cooled to room
temperature and 1N HCl was added to precipitate the product. The
solid was filtered and washed with H.sub.2O and Hexanes to give the
title compound (787 mg, 53%). 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 13.03 (s, 1H) 7.72 (dd, J=8.59, 5.81 Hz, 2H) 7.29 (t,
J=8.84 Hz, 2H) 4.31 (q, J=7.07 Hz, 2H) 1.29 (t, J=7.07 Hz, 3H).
19d) Ethyl
2-(4-biphenylylmethyl)-6-(4-fluorophenyl)-5-hydroxy-3-oxo-2,3-dihydro-4-p-
yridazinecarboxylate.
Sodium hydride (50 mg, 1.25 mmol) was added to a solution of the
compound from example 19c) (139 mg, 0.50 mmol) in
N,N-Dimethylformamide (DMF) (2.5 mL) at 0.degree. C. The reaction
was brought to room temperature and stirred for 45 minutes. The
temperature was then reduced to 0.degree. C. and
4-(bromomethyl)-biphenyl (124 mg, 0.50 mmol) was added. The
reaction was brought to room temperature and stirred for 3 hours
followed by the addition of 1N HCl. The solution was diluted with
EtOAc and H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 25-45% EtOAc/Hexanes) give the title compound (69 mg,
31%). LCMS (ES.sup.+) m/z 445.2 (MH.sup.+).
19e)
N-{[2-(4-Biphenylylmethyl)-6-(4-fluorophenyl)-5-hydroxy-3-oxo-2,3-di-
hydro-4-pyridazinyl] carbonyl}glycine. Glycine, sodium salt (28 mg,
0.29 mmol) was added to a solution of the compound from example
19d) (65 mg, 0.15 mmol) in 2-methoxyethanol (1.5 mL) at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes to give the title compound (19 mg, 28%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 13.02 (s, 1H) 10.26 (t,
J=4.93 Hz, 1H) 7.87 (dd, J=8.72, 5.68 Hz, 2H) 7.60-7.69 (m, 4H)
7.42-7.51 (m, 4H) 7.29-7.40 (m, 3 H) 5.41 (s, 2H) 4.14 (d, J=5.81
Hz, 2H).
Example 20
##STR00031##
N-[(6-(4-Chlorophenyl)-2-{[4-(1,1-dimethylethyl)phenyl]methyl}-5-hydroxy-3-
-oxo-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
20a) Ethyl
3-{2-[1-(4-chlorophenyl)-2-(ethyloxy)-2-oxo-ethylidene]hydrazino}-3-oxopr-
opanoate. A solution of diethyl oxylate (3.4 mL, 25.0 mmol) in THF
(50 mL) and Et.sub.2O (50 mmol) was cooled to -78.degree. C.
4-chlorophenyl magnesium bromide (1.0M solution in Et.sub.2O, 30
mL, 30.0 mmol) was dropwise added and the solution stirred under a
nitrogen atmosphere for 2 h at -78.degree. C. The reaction was
brought to 0.degree. C. and quenched with 6N HCl. Additional
Et.sub.2O and H.sub.2O were added and the layers separated. The
aqueous phase was backextracted with Et.sub.2O several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The resulting oil was dissolved in EtOH.
Ethyl-3-hydrazino-3-oxopropionate (4.27 g, 29.2 mmol) and catalytic
AcOH (0.3 mL, 4.87 mmol) were added along with several spatula tips
of MgSO.sub.4. The reaction was heated to reflux and stirred
overnight. The reaction was cooled and solvent removed under
reduced pressure. The residue was azeotroped with Toluene several
times. The product was purified by column chromatography
(SiO.sub.2, 15-45% EtOAc/Hexanes) to give the title compound (6.29
g, 76% over 2 steps). 1H NMR (400 MHz. DMSO-d.sub.6) .delta. ppm
10.60-11.75 (m, 1H) 7.21-7.72 (m, 4H) 3.96-4.50 (m, 4H) 3.40-3.82
(m, 2H) 1.02-1.38 (m, 6H).
20b) Ethyl
6-(4-chlorophenyl)-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylate.
KHMDS (1.80 g, 9.0 mmol) was added in several portions to a
solution of the compound from example 20a) (2.04 g, 6.0 mmol) in
1,4-dioxane at room temperature. The reaction was heated to reflux
and stirred for 5 h. The reaction was cooled back to room
temperature and 1N HCl was added to precipitate the product.
CH.sub.2Cl.sub.2 and H.sub.2O were added and the layers separated.
The aqueous phase was backextracted with CH.sub.2Cl.sub.2 several
times. The combined organic layers were washed with Brine, dried
(MgSO.sub.4), filtered and concentrated to give a pale yellow
solid. The flask was cooled and Et.sub.2O added. The product was
filtered to give the title compound as a white solid (1.07 g, 60%).
LCMS (ES.sup.+) m/z 294.8 (MH.sup.+).
20c) Ethyl 6-(4-chlorophenyl)-2-{[4-(1,1-dimethylethyl)
phenyl]methyl}-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylate.
Sodium hydride (50 mg, 1.25 mmol) was added to a solution of the
compound from example 20b) (147 mg, 0.50 mmol) in
N,N-Dimethylformamide (DMF) at 0.degree. C. The reaction was
brought to room temperature and stirred for 45 minutes. The
temperature was then reduced to 0.degree. C. and 4-tert-butylbenzyl
bromide (0.09 mL, 0.50 mmol) was added. The reaction was brought to
room temperature and stirred for 3 hours followed by the addition
of 1N HCl. The solution was diluted with EtOAc and H.sub.2O and the
layers separated. The aqueous layer was backextracted with EtOAc
several times. The combined organic layers were washed with Brine,
dried (MgSO.sub.4), filtered and concentrated. The product was
purified by column chromatography (SiO.sub.2, 1-5%
MeOH/CH.sub.2Cl.sub.2) then triturating with cold Et.sub.2O to give
the title compound as a white foam (132 mg, 60%). LCMS (ES.sup.+)
m/z 441.1 (MH.sup.+).
20d) N-[(6-(4-Chlorophenyl)-2-{[4-(1,1-dimethylethyl)
phenyl]methyl}-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinyl)carbonyl]glycine-
. Glycine, sodium salt (55 mg, 0.57 mmol) was added to a solution
of the compound from example 20c) (125 mg, 0.28 mmol) in
2-methoxyethanol at room temperature. The reaction was heated to
reflux and stirred for 2 h. The reaction was cooled back to room
temperature and H.sub.2O was added. The solution was filtered and
1N HCl was added to precipitate the product. The solid was filtered
and washed with H.sub.2O and Hexanes. The product was purified by
precipitation from CH.sub.2Cl.sub.2/Hexanes to give the title
compound as a white solid (69 mg, 52%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 10.24 (t, J=6.06 Hz, 1H) 7.84 (d, J=8.59
Hz, 2 H) 7.57 (d, J=8.59 Hz, 2H) 7.33-7.41 (m, 2H) 7.26-7.33 (m,
2H) 5.32 (s, 2H) 4.12 (d, J=5.81 Hz, 2H) 1.25 (s, 9H).
Example 21
##STR00032##
N-{[2-(4-Biphenylylmethyl)-6-(4-chlorophenyl)-5-hydroxy-3-oxo-2,3-dihydro--
4-pyridazinyl]carbonyl}glycine
21a) Ethyl
2-(4-biphenylylmethyl)-6-(4-chlorophenyl)-5-hydroxy-3-oxo-2,3-dihydro-4-p-
yridazinecarboxylate.
Sodium hydride (50 mg, 1.25 mmol) was added to a solution of the
compound from example 20b) (147 mg, 0.50 mmol) in
N,N-Dimethylformamide (DMF) (2.5 mL) at 0.degree. C. The reaction
was brought to room temperature and stirred for 45 minutes. The
temperature was then reduced to 0.degree. C. and
4-(bromomethyl)-biphenyl (124 mg, 0.50 mmol) was added. The
reaction was brought to room temperature and stirred for 3 hours
followed by the addition of 1N HCl. The solution was diluted with
EtOAc and H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 1-5% MeOH/CH.sub.2Cl.sub.2) give the title compound
(135 mg, 59%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 7.74
(ddd, J=8.91, 2.53, 2.21 Hz, 2H) 7.60-7.67 (m, 4H) 7.55 (ddd,
J=9.09, 2.53, 2.27 Hz, 2H) 7.40-7.50 (m, 4H) 7.36 (tt, J=7.33, 1.26
Hz, 1H) 5.30 (s, 2H) 4.30 (q, J=7.07 Hz, 2H) 1.28 (t, J=7.20 Hz,
3H).
21b)
N-{[2-(4-Biphenylylmethyl)-6-(4-chlorophenyl)-5-hydroxy-3-oxo-2,3-di-
hydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt (53 mg,
0.55 mmol) was added to a solution of the compound from example
21a) (126 mg, 0.27 mmol) in 2-methoxyethanol (1.5 mL) at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes. The product was purified by precipitation
from CH.sub.2Cl.sub.2/Hexanes to give the title compound (90 mg,
67%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 13.03 (s, 1H)
10.25 (t, J=5.43 Hz, 1H) 7.80-7.89 (m, 2H) 7.65 (ddd, J=6.88, 3.09,
1.64 Hz, 4H) 7.58 (ddd, J=9.09, 2.53, 2.27 Hz, 2H) 7.42-7.50 (m,
4H) 7.32-7.40 (m, 1H) 5.41 (s, 2H) 4.14 (d, J=5.56 Hz, 2H).
Example 22
##STR00033##
N-{[2-[(2-Chlorophenyl)methyl]-6-(4-fluorophenyl)-5-hydroxy-3-oxo-2,3-dihy-
dro-4-pyridazinyl]carbonyl}glycine
22a) Ethyl
2-[(2-chlorophenyl)methyl]-6-(4-fluorophenyl)-5-hydroxy-3-oxo-2,3-dihydro-
-4-pyridazinecarboxylate. Sodium hydride (83 mg, 2.08 mmol) was
added to a solution of the compound from example 19c) (232 mg, 0.83
mmol) in N,N-Dimethylformamide (DMF) (4 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 45
minutes. The temperature was then reduced to 0.degree. C. and
2-chlorobenzyl bromide (0.11 mL, 0.83 mmol) was added. The reaction
was brought to room temperature and stirred for 3 hours followed by
the addition of 1N HCl. The solution was diluted with EtOAc and
H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 1-5% MeOH/CH.sub.2Cl.sub.2) to give the title compound
(165 mg, 49%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.72
(br. s., 1H) 7.66-7.76 (m, 2H) 7.44-7.54 (m, 1H) 7.22-7.39 (m, 4H)
7.18 (dd, J=6.82, 2.27 Hz, 1H) 5.34 (s, 2H) 4.31 (q, J=7.07 Hz, 2H)
1.28 (t, J=7.07 Hz, 3H).
22b)
N-{[2-[(2-Chlorophenyl)methyl]-6-(4-fluorophenyl)-5-hydroxy-3-oxo-2,-
3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt (71
mg, 0.73 mmol) was added to a solution of the compound from example
22a) (147 mg, 0.37 mmol) in 2-methoxyethanol (2 mL) at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes. The product was redissolved in hot
CH.sub.2Cl.sub.2, dried (MgSO.sub.4) and filtered. The solution was
cooled to 0.degree. C. and Hexanes added to precipitate the
product. The product was filtered to give the title compound as a
pale pink solid (107 mg, 68%). 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 13.03 (s, 1H) 10.20 (t, J=4.29 Hz, 1H) 7.76-7.86 (m,
2H) 7.52 (dd, J=7.58, 1.52 Hz, 1H) 7.27-7.41 (m, 4H) 7.20-7.27 (m,
1H) 5.45 (s, 2H) 4.14 (d, J=5.56 Hz, 2H).
Example 23
##STR00034##
N-({6-(4-Chlorophenyl)-2-[(2-chlorophenyl)methyl]-5-hydroxy-3-oxo-2,3-dihy-
dro-4-pyridazinyl}carbonyl)glycine
23a) Ethyl
6-(4-chlorophenyl)-2-[(2-chlorophenyl)methyl]-5-hydroxy-3-oxo-2,3-dihydro-
-4-pyridazinecarboxylate. Sodium hydride (85 mg, 2.14 mmol) was
added to a solution of the compound from example 20b) (250 mg, 0.85
mmol) in N,N-Dimethylformamide (DMF) (4 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 45
minutes. The temperature was then reduced to 0.degree. C. and
2-chlorobenzyl bromide (0.11 mL, 0.85 mmol) was added. The reaction
was brought to room temperature and stirred for 3 hours followed by
the addition of 1N HCl. The solution was diluted with EtOAc and
H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 1-5% MeOH/CH.sub.2Cl.sub.2) then triturating with cold
Et.sub.2O to give the title compound as a white solid (187 mg,
53%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 7.70 (ddd, J=9.09,
2.53, 2.27 Hz, 2H) 7.47-7.55 (m, 3H) 7.28-7.38 (m, 2H) 7.14-7.20
(m, 1H) 5.34 (s, 2 H) 4.30 (q, J=7.16 Hz, 2H) 1.28 (t, J=7.07 Hz,
3H).
23b)
N-({6-(4-Chlorophenyl)-2-[(2-chlorophenyl)methyl]-5-hydroxy-3-oxo-2,-
3-dihydro-4-pyridazinyl}carbonyl)glycine. Glycine, sodium salt (83
mg, 0.85 mmol) was added to a solution of the compound from example
23a) (179 mg, 0.43 mmol) in 2-methoxyethanol at room temperature.
The reaction was heated to reflux and stirred for 2 h. The reaction
was cooled back to room temperature and H.sub.2O was added. The
solution was filtered and 1N HCl was added to precipitate the
product. The solid was filtered and washed with H.sub.2O and
Hexanes to give the title compound (153 mg, 80%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 13.03 (s, 1H) 10.19 (t, J=4.80 Hz, 1H)
7.74-7.83 (m, 2H) 7.55 (ddd, J=8.91, 2.53, 2.21 Hz, 2H) 7.52 (dd,
J=7.71, 1.39 Hz, 1H) 7.28-7.39 (m, 2H) 7.21-7.26 (m, 1H) 5.45 (s,
2H) 4.13 (d, J=5.81 Hz, 2H).
Example 24
##STR00035##
N-{[6-(4-Chlorophenyl)-2-(2-cyclohexylethyl)-5-hydroxy-3-oxo-2,3-dihydro-4-
-pyridazinyl]carbonyl}glycine
24a) Ethyl
6-(4-chlorophenyl)-2-(2-cyclohexylethyl)-5-hydroxy-3-oxo-2,3-dihydro-4-py-
ridazinecarboxylate.
Sodium hydride (85 mg, 2.14 mmol) was added to a solution of the
compound from example 20b) (250 mg, 0.85 mmol) in
N,N-Dimethylformamide (DMF) (4 mL) at 0.degree. C. The reaction was
brought to room temperature and stirred for 45 minutes. The
temperature was then reduced to 0.degree. C. and 2-cyclohexyl ethyl
bromide (0.13 mL, 0.85 mmol) was added. The reaction was brought to
room temperature and stirred for 3 hours followed by the addition
of 1N HCl. The solution was diluted with EtOAc and H.sub.2O and the
layers separated. The aqueous layer was backextracted with EtOAc
several times. The combined organic layers were washed with Brine,
dried (MgSO.sub.4), filtered and concentrated. The product was
purified by column chromatography (SiO.sub.2, 1-5%
MeOH/CH.sub.2Cl.sub.2) then dried under high vacuum to give the
title compound as a pale yellow solid (187 mg, 54%). LCMS
(ES.sup.+) m/z 405.0 (MH.sup.+).
24b)
N-{[6-(4-Chlorophenyl)-2-(2-cyclohexylethyl)-5-hydroxy-3-oxo-2,3-dih-
ydro-4-pyridazinyl] carbonyl}glycine. Glycine, sodium salt (85 mg,
0.87 mmol) was added to a solution of the compound from example
24a) (177 mg, 0.44 mmol) in 2-methoxyethanol (2 mL) at room
temperature. The reaction was heated to reflux and stirred for 3 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes to give the title compound (117 mg, 62%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 13.02 (s, 1H) 10.32 (t,
J=4.93 Hz, 1H) 7.82 (d, J=8.59 Hz, 2H) 7.57 (ddd, J=8.84, 2.53,
2.27 Hz, 2H) 4.16-4.23 (m, 2H) 4.15 (d, J=5.81 Hz, 2H) 1.55-1.81
(m, 7H) 1.10-1.41 (m, 4H) 0.84-1.02 (m, 2 H).
Example 25
##STR00036##
N-{[2-(2-Cyclopropylethyl)-6-(4-fluorophenyl)-5-hydroxy-3-oxo-2,3-dihydro--
4-pyridazinyl]carbonyl}glycine
25a) Ethyl 2-(2-cyclopropylethyl)-6-(4-fluorophenyl)-5-hydroxy-3
-oxo-2,3-dihydro-4-pyridazinecarboxylate.
Sodium hydride (40 mg, 1.00 mmol) was added to a solution of the
compound from example 19c) (139 mg, 0.50 mmol) in
N,N-Dimethylformamide (DMF) (2.5 mL) at 0.degree. C. The reaction
was brought to room temperature and stirred for 45 minutes. The
temperature was then reduced to 0.degree. C. and cyclopropylethyl
iodide (119 mg, 0.50 mmol) was added. The reaction was brought to
room temperature and stirred for 3 hours followed by the addition
of 1N HCl. The solution was diluted with EtOAc and H.sub.2O and the
layers separated. The aqueous layer was backextracted with EtOAc
several times. The combined organic layers were washed with Brine,
dried (MgSO.sub.4), filtered and concentrated. The product was
purified by column chromatography (SiO.sub.2, 1-5%
MeOH/CH.sub.2Cl.sub.2) to give the title compound (47 mg, 27%).
LCMS (ES.sup.+) m/z 346.8 (MH.sup.+).
25b)
N-{[2-(2-Cyclopropylethyl)-6-(4-fluorophenyl)-5-hydroxy-3-oxo-2,3-di-
hydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt (26 mg,
0.27 mmol) was added to a solution of the compound from example
25a) (47 mg, 0.14 mmol) in 2-methoxyethanol (0.8 mL) at room
temperature. The reaction was heated to reflux and stirred for 3 h.
The reaction was cooled back to room temperature and 1N HCl was
added to precipitate the product. The solid was filtered and washed
with Hexanes and Et.sub.2O to give the title compound as a pale
pink solid (21 mg, 40%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
13.02 (s, 1H) 10.28-10.39 (m, 1H) 7.79-7.88 (m, 2H) 7.29-7.38 (m,
2H) 4.25 (t, J=7.07 Hz, 2H) 4.15 (d, J=5.56 Hz, 2H) 1.68 (q, J=7.07
Hz, 2H) 0.65-0.79 (m, 1H) 0.40 (ddd, J=7.89, 5.75, 4.04 Hz, 2H)
0.02 (td, J=5.24, 4.17 Hz, 2H).
Example 26
##STR00037##
N-[(6-[4-(1,1-Dimethylethyl)phenyl]-2-{[4-(1,1-dimethylethyl)phenyl]methyl-
}-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
26a) Ethyl[4-(1,1-dimethylethyl)phenyl](oxo)acetate. To a solution
of 4-bromo-tert-butylbenzene (0.70 mL, 5.0 mmol) in THF (20 mL) was
dropwise added n-BuLi (2.87 M solution in Hexanes, 1.74 mL, 5.0
mmol) at -78.degree. C. under a nitrogen atmosphere. After stirring
for 1 h diethyl oxylate (0.68 mL, 5.0 mmol) was added. The reaction
stirred for 1 h at -78.degree. C. and then 1 N HCl was added. The
solution was diluted with H.sub.2O and EtOAc and the layers
separated. The aqueous phase was backextracted with EtOAc several
times. The combined organic layers were washed with Brine, dried
(MgSO.sub.4), filtered and concentrated. The product was purified
by column chromatography (SiO.sub.2, 5-10% EtOAc/Hexanes) to give
the title compound (334 mg, 29%). 1H NMR (400 MHz, CHLOROFORM-d)
.delta. ppm 7.97 (dt, J=8.65, 2.12 Hz, 2H) 7.55 (dt, J=8.84, 2.02
Hz, 2H) 4.47 (q, J=7.07 Hz, 2H) 1.45 (t, J=7.20 Hz, 3H) 1.37 (s,
9H).
26b) Ethyl
3-{2-[1-[4-(1,1-dimethylethyl)phenyl]-2-(ethyloxy)-2-oxoethylidene]hydraz-
ino}-3-oxopropanoate.
Ethyl-3-hydrazino-3-oxopropionate (419 mg, 2.87 mmol) and catalytic
AcOH (0.03 mL, 0.48 mmol) were added to a solution of the compound
from example 26a) (560 mg, 2.39 mmol) in EtOH (10 mL). Several
spatula tips of MgSO.sub.4 were added and the reaction stirred at
reflux overnight. The reaction was then cooled to room temperature
and filtered. The filtrate was concentrated and azeotroped with
Toluene several times. The product was purified by column
chromatography (SiO.sub.2, 10-45% EtOAc/Hexanes) to give the title
compound (581 mg, 67%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
10.39-11.61 (m, 1H) 7.17-7.59 (m, 4H) 3.97-4.48 (m, 4 H) 50
3.45-3.76 (m, 2H) 1.11-1.36 (m, 15H).
26c) Ethyl
6-[4-(1,1-dimethylethyl)phenyl]-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinec-
arboxylate. KHMDS (466 mg, 2.33 mmol) was added in several portions
to a solution of the compound from example 26b) (564 mg, 1.56 mmol)
in 1,4-dioxane (4 mL) at room temperature. The reaction was heated
to reflux and stirred for 3 h. The reaction was cooled to room
temperature and solvent removed under reduced pressure. 1N HCl was
added to precipitate the product. The solid was filtered and washed
with H.sub.2O, Hexanes and Et.sub.2O to give the title compound
(220 mg, 45%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 13.03 (s,
1H) 12.59 (br. s., 1H) 7.57-7.62 (m, 2H) 7.47 (dt, J=8.59, 2.02 Hz,
2H) 4.32 (q, J=7.07 Hz, 2H) 1.31 (s, 9H) 1.26-1.34 (m, 3H).
26d) Ethyl
6-[4-(1,1-dimethylethyl)phenyl]-2-{[4-(1,1-dimethylethyl)phenyl]methyl}-5-
-hydroxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride
(32 mg, 0.79 mmol) was added to a solution of the compound from
example 26c) (100 mg, 0.32 mmol) in N,N-Dimethylformamide (DMF) at
0.degree. C. The reaction was brought to room temperature and
stirred for 45 minutes. The temperature was then reduced to
0.degree. C. and 4-tert-butylbenzyl bromide (0.06 mL, 0.32 mmol)
was added. The reaction was brought to room temperature and stirred
for 3 hours followed by the addition of 1N HCl. The solution was
diluted with EtOAc and H.sub.2O and the layers separated. The
aqueous layer was back-extracted with EtOAc several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The product was purified by column
chromatography (SiO.sub.2, 20-45% EtOAc/Hexanes) give the title
compound as a white foam (110 mg, 75%). LCMS (ES.sup.+) m/z 463.2
(MH.sup.+).
26e)
N-[(6-[4-(1,1-Dimethylethyl)phenyl]-2-{[4-(1,1-dimethylethyl)phenyl]-
methyl}-5-hydroxy-3-oxo-2,3-dihydro-4
-pyridazinyl)carbonyl]glycine. Glycine, sodium salt (43 mg, 0.44
mmol) was added to a solution of the compound from example 26d)
(102 mg, 0.22 mmol) in 2-methoxyethanol (1.4 mL) at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes which partially dissolved the product. EtOAc
was added and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated to give the title compound as a tan solid (57 mg,
53%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 16.29 (s, 1H)
13.00 (s, 1H) 10.27 (t, J=5.56 Hz, 1H) 7.72 (d, J=8.59 Hz, 2H) 7.51
(d, J=8.59 Hz, 2H) 7.34-7.41 (m, 2 H) 7.27-7.33 (m, 2 H) 5.31 (s,
2H) 4.13 (d, J=5.81 Hz, 2H) 1.32 (s, 9H) 1.25 (s, 9H).
Example 27
##STR00038##
N-({2-[(2-Chlorophenyl)methyl]-6-[4-(1,1-dimethylethyl)phenyl]-5-hydroxy-3-
-oxo-2,3-dihydro-4-pyridazinyl}carbonyl)glycine
27a) Ethyl
2-[(2-chlorophenyl)methyl]-6-[4-(1,1-dimethylethyl)phenyl]-5-hydroxy-3-ox-
o-2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride (37 mg, 0.92
mmol) was added to a solution of the compound from example 26c)
(117 mg, 0.37 mmol) in N,N-Dimethylformamide (DMF) (1.5 mL) at
0.degree. C. The reaction was brought to room temperature and
stirred for 45 minutes. The temperature was then reduced to
0.degree. C. and 2-chlorobenzyl bromide (0.05 mL, 0.37 mmol) was
added. The reaction was brought to room temperature and stirred for
3 hours followed by the addition of 1N HCl. The solution was
diluted with EtOAc and H.sub.2O and the layers separated. The
aqueous layer was backextracted with EtOAc several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The product was purified by column
chromatography (SiO.sub.2, 0-4% MeOH/CH.sub.2Cl.sub.2) then dried
under high vacuum give the title compound as a white foam (125 mg,
77%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.64 (br. s., 1H)
7.59 (ddd, J=8.72, 2.15, 2.02 Hz, 2H) 7.43-7.52 (m, 3H) 7.28-7.38
(m, 2 H) 7.13-7.21 (m, 1H) 5.34 (s, 2H) 4.31 (q, J=7.07 Hz, 2H)
1.30 (s, 9H) 1.24-1.32 (m, 3H).
27b)
N-({2-[(2-Chlorophenyl)methyl]-6-[4-(1,1-dimethylethyl)phenyl]-5-hyd-
roxy-3-oxo-2,3-dihydro-4-pyridazinyl}carbonyl)glycine. Glycine,
sodium salt (52 mg, 0.54 mmol) was added to a solution of the
compound from example 27a) (118 mg, 0.27 mmol) in 2-methoxyethanol
at room temperature. The reaction was heated to reflux and stirred
for 2 h. The reaction was cooled back to room temperature and
H.sub.2O was added. The solution was filtered and 1N HCl was added
to precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes. The product was purified by precipitation
from CH.sub.2Cl.sub.2/Hexanes to give the title compound as a white
solid (100 mg, 79%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
13.01 (s, 1H) 10.22 (t, J=5.18 Hz, 1H) 7.64-7.71 (m, 2H) 7.46-7.54
(m, 3H) 7.28-7.40 (m, 2H) 7.20-7.26 (m, 1H) 5.45 (s, 2H) 4.14 (d,
J=5.56 Hz, 2H) 1.30 (s, 9H).
Example 28
##STR00039##
N-({5-Hydroxy-6-methyl-2-[(2-nitrophenyl)methyl]-3-oxo-2,3-dihydro-4-pyrid-
azinyl}carbonyl)glycine
28a) Ethyl
3-{(2Z)-2-[2-(ethyloxy)-1-methyl-2-oxoethylidene]hydrazino}-3-oxopropanoa-
te. Ethyl-3-hydrazino-3-oxopropionate (0.73 g, 5.00 mmol) and
catalytic AcOH (0.06 mL, 1.05 mmol) were added to a solution of
ethyl pyruvate (0.55 mL, 5.00 mmol) in CH.sub.2Cl.sub.2 (20 mL).
The reaction stirred overnight at room temperature. The solvent was
removed under reduced pressure and the resulting solid was
azeotroped with Toluene several times. The product was purified by
column chromatography (SiO.sub.2, 1-5% MeOH/CH.sub.2Cl.sub.2) to
give the title compound as a white solid (1.29 g, 63%). 1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 10.64-11.20 (m, 1H) 4.03-4.28 (m, 4
H) 3.47-3.68 (m, 2H) 1.96-2.11 (m, 3H) 1.12-1.32 (m, 6H).
28b) Ethyl
5-hydroxy-6-methyl-3-oxo-2,3-dihydro-4-pyridazinecarboxylate. DBU
(1.6 mL, 10.73 mmol) was dropwise added to a solution of the
compound from example 28a) (1.31 g, 5.36 mmol) in 1,4-dioxane (12
mL) at room temperature. The reaction was heated to reflux and
stirred overnight. The reaction was cooled to room temperature and
solvent removed under reduced pressure. The product was purified by
pushing through a silica plug with MeOH to give the title compound
(348 mg, 33%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 1.27 (t,
J=7.20 Hz, 3H) 2.17 (s, 3H) 4.27 (q, J=7.24 Hz, 2H) 12.13 (br. s.,
1H) 12.59 (s, 1H).
28c) Ethyl
5-hydroxy-6-methyl-2-[(2-nitrophenyl)methyl]-3-oxo-2,3-dihydro-4-pyridazi-
necarboxylate. Sodium hydride (50 mg, 1.26 mmol) was added to a
solution of the compound from example 28b) (100 mg, 0.50 mmol) in
N,N-Dimethylformamide (DMF) (2 mL) at 0.degree. C. The reaction was
brought to room temperature and stirred for 45 minutes. The
temperature was then reduced to 0.degree. C. and 2-nitrobenzyl
bromide (109 mg, 0.50 mmol) was added. The reaction was brought to
room temperature and stirred for 3 hours followed by the addition
of 1N HCl. The solution was diluted with EtOAc and H.sub.2O and the
layers separated. The aqueous layer was backextracted with EtOAc
several times. The combined organic layers were washed with Brine,
dried (MgSO.sub.4), filtered and concentrated. The product was
purified by column chromatography (SiO.sub.2, 0-6%
MeOH/CH.sub.2Cl.sub.2) to give the title compound as an orange oil
(83 mg, 50%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.28 (s,
1H) 8.08 (dd, J=8.08, 1.26 Hz, 1H) 7.70 (td, J=7.64, 1.39 Hz, 1H)
7.53-7.61 (m, 1H) 7.18 (dd, J=7.71, 0.88 Hz, 1H) 5.45 (s, 2H) 4.26
(q, J=7.24 Hz, 2H) 2.20 (s, 3H) 1.25 (t, J=7.07 Hz, 3H).
28d)
N-({5-Hydroxy-6-methyl-2-[(2-nitrophenyl)methyl]-3-oxo-2,3-dihydro-4-
-pyridazinyl}carbonyl)glycine.
Glycine, sodium salt (43 mg, 0.45 mmol) was added to a solution of
the compound from example 28c) (75 mg, 0.23 mmol) in
2-methoxyethanol at room temperature. The reaction was heated to
reflux and stirred for 2 h. The reaction was cooled back to room
temperature and H.sub.2O was added. The solution was filtered and
1N HCl was added to precipitate the product. The solid was filtered
and washed with H.sub.2O and Hexanes. The product was purified by
precipitation from CH.sub.2Cl.sub.2/Hexanes to give the title
compound as a brown solid (20 mg, 24%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 15.75 (s, 1H) 12.99 (s, 1H) 10.03 (t,
J=5.18 Hz, 1H) 8.11 (dd, J=8.08, 1.26 Hz, 1H) 7.69 (td, J=7.64,
1.39 Hz, 1H) 7.59 (td, J=7.71, 1.26 Hz, 1H) 7.23 (dd, J=7.83, 1.01
Hz, 1H) 5.56 (s, 2H) 4.09 (d, J=5.81 Hz, 2H) 2.24 (s, 3H).
Example 29
##STR00040##
N-[(6-(4-Fluorophenyl)-5-hydroxy-3-oxo-2-{[2-(trifluoromethyl)phenyl]methy-
l}-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
29a) Ethyl
6-(4-fluorophenyl)-5-hydroxy-3-oxo-2-{[2-(trifluoromethyl)phenyl]methyl}--
2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride (40 mg, 1.01
mmol) was added to a solution of the compound from example 19c)
(112 mg, 0.40 mmol) in N,N-Dimethylformamide (DMF) at 0.degree. C.
The reaction was brought to room temperature and stirred for 45
minutes. The temperature was then reduced to 0.degree. C. and
2-(trifluoromethyl)-benzyl bromide (96 mg, 0.40 mmol) was added.
The reaction was brought to room temperature and stirred for 3
hours followed by the addition of 1N HCl. The solution was diluted
with EtOAc and H.sub.2O and the layers separated. The aqueous layer
was backextracted with EtOAc several times. The combined organic
layers were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 1-5% MeOH/CH.sub.2Cl.sub.2) to give the title compound
(170 mg, 97%). LCMS (ES.sup.+) m/z 437.0 (MH.sup.+).
29b)
N-[(6-(4-Fluorophenyl)-5-hydroxy-3-oxo-2-{[2-(trifluoromethyl)phenyl-
]methyl}-2,3-dihydro-4-pyridazinyl) carbonyl]glycine. Glycine,
sodium salt (68 mg, 0.70 mmol) was added to a solution of the
compound from example 29a) (153 mg, 0.35 mmol) in 2-methoxyethanol
(1.5 mL) at room temperature. The reaction was heated to reflux and
stirred for 2 h. The reaction was cooled back to room temperature
and H.sub.2O was added. The solution was filtered and 1N HCl was
added to precipitate the product. The solid was filtered and washed
with H.sub.2O, Hexanes and Et.sub.2O to give the title compound as
a pale pink solid (71 mg, 44%). 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 15.75 (s, 1H) 12.99 (s, 1H) 10.03 (t, J=5.18 Hz, 1H)
8.11 (dd, J=8.08, 1.26 Hz, 1H) 7.69 (td, J=7.64, 1.39 Hz, 1H) 7.59
(td, J=7.71, 1.26 Hz, 1H) 7.23 (dd, J=7.83, 1.01 Hz, 1H) 5.56 (s, 2
H) 4.09 (d, J=5.81 Hz, 2H) 2.24 (s, 3H).
Example 30
##STR00041##
N-{[2-[(2-Chlorophenyl)methyl]-6-(3,5-difluorophenyl)-5-hydroxy-3-oxo-2,3--
dihydro-4-pyridazinyl]carbonyl}glycine
30a) Ethyl
3-{2-[1-(3,5-difluorophenyl)-2-(ethyloxy)-2-oxoethylidene]hydrazino}-3-ox-
opropanoate. A solution of diethyl oxylate (2.0 mL, 15.0 mmol) in
THF (30 mL) and Et.sub.2O (30 mmol) was cooled to -78.degree. C.
3,5-difluorophenyl magnesium bromide (0.5 M solution in THF, 36 mL,
18.0 mmol) was dropwise added and the solution stirred under a
nitrogen atmosphere for 1.5 h at -78.degree. C. The reaction was
brought to 0.degree. C. and quenched with 6N HCl. Additional
Et.sub.2O and H.sub.2O were added and the layers separated. The
aqueous phase was backextracted with Et.sub.2O several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The product was purified by column
chromatography (SiO.sub.2, 15-45% EtOAc/Hexanes). The crude oil was
redissolved in EtOH (40 mL). Ethyl-3-hydrazino-3-oxopropionate
(2.63 g, 18.0 mmol) and catalytic AcOH (0.2 mL, 3.00 mmol) were
added along with a few spatula tips of MgSO.sub.4. The reaction was
heated to reflux and stirred overnight. The reaction was then
cooled to room temperature and filtered. The filtrate was
concentrated and azeotroped with Toluene several times. The product
was purified by column chromatography (SiO.sub.2, 10-40%
EtOAc/Hexanes) to give the title compound (1.01 g, 20% over 2
steps). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 10.62-11.95 (m,
1H) 6.85-7.50 (m, 3H) 3.99-4.51 (m, 4H) 3.42-3.86 (m, 2H) 0.95-1.41
(m, 6H).
30b) Ethyl
6-(3,5-difluorophenyl)-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylat-
e. KHMDS (0.80 g, 4.01 mmol) was added in several portions to a
solution of the compound from example 30a) (0.915 g, 2.67 mmol) in
1,4-dioxane (15 mL) at room temperature. The reaction was heated to
reflux and stirred for 1.5 h. The reaction was cooled to room
temperature and diluted with EtOAc and H.sub.2O. 1N HCl was added
to neutralize the solution. The layers were separated and the
aqueous phase was backextracted with EtOAc several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated to give a pale yellow residue. The flask
was cooled and Et.sub.2O was added. The product was filtered to
give the title compound as a white solid (332 mg, 42%). 1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 13.19 (s, 1H) 7.31-7.45 (m, 3H) 4.31
(q, J=7.07 Hz, 2H) 1.29 (t, J=7.07 Hz, 3H).
30c) Ethyl
2-[(2-chlorophenyl)methyl]-6-(3,5-difluorophenyl)-5-hydroxy-3-oxo-2,3-dih-
ydro-4-pyridazinecarboxylate. Sodium hydride (38 mg, 0.95 mmol) was
added to a solution of the compound from example 30b) (113 mg, 0.38
mmol) in N,N-Dimethylformamide (DMF) (1.8 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 45
minutes. The temperature was then reduced to 0.degree. C. and
2-chlorobenzyl bromide (0.05 mL, 0.38 mmol) was added. The reaction
was brought to room temperature and stirred for 3 hours followed by
the addition of 1N HCl. The solution was diluted with EtOAc and
H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 40-65% EtOAc/Hexanes) give the title compound (124 mg,
78%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 7.47-7.55 (m, 1H)
7.28-7.45 (m, 5H) 7.16-7.24 (m, 1H) 5.36 (s, 2H) 4.30 (q, J=7.24
Hz, 2H) 1.28 (t, J=7.07 Hz, 3H).
30d)
N-{[2-[(2-Chlorophenyl)methyl]-6-(3,5-difluorophenyl)-5-hydroxy-3-ox-
o-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt
(48 mg, 0.50 mmol) was added to a solution of the compound from
example 30c) (105 mg, 0.25 mmol) in 2-methoxyethanol (1.5 mL) at
room temperature. The reaction was heated to reflux and stirred for
2 h. The reaction was cooled back to room temperature and H.sub.2O
was added. The solution was filtered and 1N HCl was added to
precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes to give the title compound (81 mg, 72%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 13.03 (s, 1H) 10.13-10.21
(m, 1H) 7.47-7.55 (m, 3H) 7.37-7.44 (m, 1H) 7.34 (ddd, J=14.78,
7.45, 1.77 Hz, 2 H) 7.23-7.29 (m, 1H) 5.47 (s, 2H) 4.13 (d, J=5.56
Hz, 2H).
Example 31
##STR00042##
N-({2-[(2-Chlorophenyl)methyl]-5-hydroxy-6-[4-(methyloxy)phenyl]-3-oxo-2,3-
-dihydro-4-pyridazinyl}carbonyl)glycine
31a) Ethyl[4-(methyloxy)phenyl](oxo)acetate. A solution of diethyl
oxylate (2.0 mL, 15.0 mmol) in THF (40 mL) and Et.sub.2O (40 mmol)
was cooled to -78.degree. C. 4-methoxyphenyl magnesium bromide (0.5
M solution in THF, 36 mL, 18.0 mmol) was dropwise added and the
solution stirred under a nitrogen atmosphere for 1.5 h at
-78.degree. C. The reaction was brought to 0.degree. C. and
quenched with 6N HCl. Additional Et.sub.2O and H.sub.2O were added
and the layers separated. The aqueous phase was backextracted with
Et.sub.2O several times. The combined organic layers were washed
with Brine, dried (MgSO.sub.4), filtered and concentrated. The
product was purified by column chromatography (SiO.sub.2, 10-20%
EtOAc/Hexanes) to give the title compound as a pale yellow oil
(2.68 g, 84%). 1H NMR (400 MHz, CHLOROFORM-d) .delta. ppm 8.02
(ddd, J=9.47, 2.78, 2.40 Hz, 2H) 7.00 (ddd, J=9.47, 2.78, 2.40 Hz,
2H) 4.46 (q, J=7.24 Hz, 2H) 3.92 (s, 3H) 1.44 (t, J=7.07 Hz,
3H).
31b) Ethyl
3-(2-{2-(ethyloxy)-1-[4-(methyloxy)phenyl]-2-oxoethylidene}hydrazino)-3-o-
xopropanoate. Ethyl-3-hydrazino-3-oxopropionate (1.24 g, 8.49 mmol)
and catalytic AcOH (0.1 mL, 1.70 mmol) were added to a solution of
the compound from example 31a) (2.38 g. 11.43 mmol) in EtOH (20
mL). The reaction was heated to reflux and stirred overnight. A few
spatula tips of MgSO.sub.4 were added and the reaction continued to
stir for 3 h. The reaction was then cooled to room temperature and
filtered. The filtrate was concentrated and azeotroped with Toluene
several times. The product was purified by column chromatography
(SiO.sub.2, 15-45% EtOAc/Hexanes) to give the title compound (2.24
g, 78%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 10.48-11.47 (m,
1H) 7.19-7.61 (m, 2H) 6.94-7.12 (m, 2H) 4.02-4.48 (m, 4H) 3.42-3.86
(m, 5H) 1.09-1.36 (m, 6H).
31c) Ethyl
5-hydroxy-6-[4-(methyloxy)phenyl]-3-oxo-2,3-dihydro-4-pyridazinecarboxyla-
te. KHMDS (1.82 g, 9.14 mmol) was added in several portions to a
solution of the compound from example 31b) (2.05 g, 6.10 mmol) in
1,4-dioxane (15 mL) at room temperature. The reaction was heated to
reflux and stirred for 3 h. The reaction was cooled to room
temperature and 1N HCl was added to precipitate the product.
CH.sub.2Cl.sub.2 and H.sub.2O were added and the layers separated.
The aqueous phase was backextracted with CH.sub.2Cl.sub.2 several
times. The combined organic layers were washed with Brine, dried
(MgSO.sub.4), filtered and concentrated. Et.sub.2O was added and
the product filtered to give the title compound as a pale yellow
solid (1.14 g, 64%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
12.99 (s, 1H) 12.62 (br. s., 1H) 7.55-7.65 (m, 2H) 6.93-7.06 (m,
2H) 4.32 (q, J=7.07 Hz, 2H) 3.80 (s, 3H) 1.29 (t, J=7.20 Hz,
3H).
31d) Ethyl
2-[(2-chlorophenyl)methyl]-5-hydroxy-6-[4-(methyloxy)phenyl]-3-oxo-2,3-di-
hydro-4-pyridazinecarboxylate. Sodium hydride (50 mg, 1.25 mmol)
was added to a solution of the compound from example 31c) (145 mg,
0.50 mmol) in N,N-Dimethylformamide (DMF) (2 mL) at 0.degree. C.
The reaction was brought to room temperature and stirred for 45
minutes. The temperature was then reduced to 0.degree. C. and
2-chlorobenzyl bromide (0.06 mL, 0.50 mmol) was added. The reaction
was brought to room temperature and stirred for 2.5 h followed by
the addition of 1N HCl. The solution was diluted with EtOAc and
H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 1-5% MeOH/CH.sub.2Cl.sub.2 then 40-80% EtOAc/Hexanes)
give the title compound (106 mg, 51%). 1H NMR (400 MHz, MeOD)
.delta. ppm 7.69-7.77 (m, 2H) 7.42-7.47 (m, 1H) 7.25-7.34 (m, 2H)
7.18-7.24 (m, 1H) 6.92-7.00 (m, 2H) 5.49 (s, 2H) 4.49 (q, J=7.07
Hz, 2H) 3.84 (s, 3 H) 1.43 (t, J=7.07 Hz, 3H).
31e)
N-({2-[(2-Chlorophenyl)methyl]-5-hydroxy-6-[4-(methyloxy)phenyl]-3-o-
xo-2,3-dihydro-4-pyridazinyl}carbonyl)glycine. Glycine, sodium salt
(40 mg, 0.41 mmol) was added to a solution of the compound from
example 31d) (85 mg, 0.21 mmol) in 2-methoxyethanol (1.5 mL) at
room temperature. The reaction was heated to reflux and stirred for
2 h. The reaction was cooled back to room temperature and H.sub.2O
was added. The solution was filtered and 1N HCl was added to
precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes. The product was purified by precipitation
from CH.sub.2Cl.sub.2/Hexanes to give the title compound (59 mg,
65%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 13.03 (s, 1H)
10.24 (t, J=4.93 Hz, 1H) 7.67-7.77 (m, 2H) 7.51 (dd, J=7.71, 1.64
Hz, 1H) 7.28-7.39 (m, 2H) 7.23 (dd, J=7.45, 1.89 Hz, 1H) 7.03 (ddd,
J=9.41, 2.78, 2.46 Hz, 2H) 5.44 (s, 2H) 4.13 (d, J=5.56 Hz, 2H)
3.80 (s, 3H).
Example 32
##STR00043##
N-({2-[(3,5-Difluorophenyl)methyl]-5-hydroxy-6-[4-(methyloxy)phenyl]-3-oxo-
-2,3-dihydro-4-pyridazinyl}carbonyl)glycine
32a) Ethyl
2-[(3,5-difluorophenyl)methyl]-5-hydroxy-6-[4-(methyloxy)phenyl]-3-oxo-2,-
3-dihydro-4-pyridazinecarboxylate. Sodium hydride (91 mg, 2.28
mmol) was added to a solution of the compound from example 31c)
(265 mg, 0.91 mmol) in N,N-Dimethylformamide (DMF) (3.5 mL) at
0.degree. C. The reaction was brought to room temperature and
stirred for 45 minutes. The temperature was then reduced to
0.degree. C. and 3,5-difluorobenzyl bromide (0.13 mL, 1.00 mmol)
was added. The reaction was brought to room temperature and stirred
for 3.5 h followed by the addition of 1N HCl. The solution was
diluted with EtOAc and H.sub.2O and the layers separated. The
aqueous layer was backextracted with EtOAc several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The product was purified by column
chromatography (SiO.sub.2, 40-65% EtOAc/Hexanes) give the title
compound (247 mg, 65%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
12.65 (br. s., 1H) 7.64 (ddd, J=9.35, 2.78, 2.53 Hz, 2H) 7.18 (tt,
J=9.44, 2.31 Hz, 1H) 6.98-7.08 (m, 4H) 5.26 (s, 2H) 4.31 (q, J=7.24
Hz, 2H) 3.80 (s, 3 H) 1.29 (t, J=7.07 Hz, 3H).
32b)
N-({2-[(3,5-Difluorophenyl)methyl]-5-hydroxy-6-[4-(methyloxy)phenyl]-
-3-oxo-2,3-dihydro-4-pyridazinyl}carbonyl)glycine. Glycine, sodium
salt (112 mg, 1.16 mmol) was added to a solution of the compound
from example 32a) (241 mg, 0.58 mmol) in 2-methoxyethanol (1.5 mL)
at room temperature. The reaction was heated to reflux and stirred
for 2 h. The reaction was cooled back to room temperature and
H.sub.2O was added. The solution was filtered and 1N HCl was added
to precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes to give the title compound (220 mg, 85%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 13.08 (br. s., 1H) 10.23
(t, J=5.43 Hz, 1H) 7.72-7.78 (m, 2H) 7.18 (tt, J=9.38, 2.37 Hz, 1H)
7.01-7.12 (m, 4H) 5.37 (s, 2H) 4.13 (d, J=5.56 Hz, 2H) 3.81 (s,
3H).
Example 33
##STR00044##
N-{[2-[(2-Chlorophenyl)methyl]-5-hydroxy-6-(4-hydroxyphenyl)-3-oxo-2,3-dih-
ydro-4-pyridazinyl]carbonyl}glycine
A solution of the compound from example 31e) in glacial acetic acid
(1.6 mL) was heated to reflux followed by the addition of 48%
aqueous HBr (0.4 mL). After stirring for 6.5 h at reflux, the
reaction was cooled to room temperature and H.sub.2O was added. The
precipitate was filtered to give the title compound as a tan solid
(35 mg, 54%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 16.41 (s,
1H) 13.00 (s, 1H) 10.24 (t, J=5.56 Hz, 1H) 9.85 (s, 1H) 7.62 (d,
J=8.59 Hz, 2H) 7.51 (dd, J=7.71, 1.64 Hz, 1H) 7.26-7.39 (m, 2H)
7.20 (dd, J=7.45, 1.89 Hz, 1H) 6.80-6.87 (m, 2H) 5.43 (s, 2H) 4.13
(d, J=5.56 Hz, 2H).
Example 34
##STR00045##
N-{[2-[(3,5-Difluorophenyl)methyl]-5-hydroxy-6-(4-hydroxyphenyl)-3-oxo-2,3-
-dihydro-4-pyridazinyl]carbonyl}glycine
A solution of the compound from example 32b) in glacial acetic acid
(2.6 mL) was heated to reflux followed by the addition of 48%
aqueous HBr (0.6 mL). After stirring overnight at reflux, the
reaction was cooled to room temperature and H.sub.2O was added. The
precipitate was filtered and washed with H.sub.2O and Hexanes to
give the title compound (70 mg, 78%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 16.37 (s, 1H) 13.00 (s, 1H) 10.23 (t,
J=5.56 Hz, 1 H) 9.86 (s, 1H) 7.65 (d, J=8.59 Hz, 2H) 7.18 (tt,
J=9.44, 2.31 Hz, 1H) 7.07 (ddd, J=14.84, 6.63, 2.27 Hz, 2H) 6.85
(ddd, J=9.22, 2.78, 2.40 Hz, 2H) 5.36 (s, 2H) 4.13 (d, J=5.81 Hz,
2H).
Example 35
##STR00046##
N-[(5-Hydroxy-6-[3-(methyloxy)phenyl]-3-oxo-2-{[2-(trifluoromethyl)phenyl]-
methyl}-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
35a) Ethyl
3-(2-{2-(ethyloxy)-1-[3-(methyloxy)phenyl]-2-oxoethylidene}hydrazino)-3-o-
xopropanoate. A solution of diethyl oxylate (2.0 mL, 15.0 mmol) in
THF (30 mL) and Et.sub.2O (30 mmol) was cooled to -78.degree. C.
3-methoxyphenyl magnesium bromide (1.0 M solution in THF, 18 mL,
18.0 mmol) was dropwise added and the solution stirred under a
nitrogen atmosphere for 2 h at -78.degree. C. The reaction was
brought to 0.degree. C. and quenched with 6N HCl. Additional
Et.sub.2O and H.sub.2O were added and the layers separated. The
aqueous phase was backextracted with Et.sub.2O several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The resulting residue was dissolved in
EtOH (40 mL). Ethyl-3-hydrazino-3-oxopropionate (2.63 g, 18.0 mmol)
and catalytic AcOH (0.2 mL, 3.49 mmol) were added along with a few
spatula tips of MgSO4. The reaction was heated to reflux and
stirred overnight. The reaction was cooled to room temperature and
filtered. The filtrate was concentrated and azeotroped with Toluene
several times. The product was purified by column chromatography
(SiO.sub.2, 15-45% EtOAc/Hexanes) to give the title compound (3.56
g, 70% over 2 steps). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
10.53-11.65 (m, 1H) 7.27-7.49 (m, 1H) 6.67-7.21 (m, 3H) 4.00-4.50
(m, 4H) 3.45-3.85 (m, 5H) 1.08-1.38 (m, 6H).
35b) Ethyl
5-hydroxy-6-[3-(methyloxy)phenyl]-3-oxo-2,3-dihydro-4-pyridazinecarboxyla-
te. KHMDS (2.67 g, 13.38 mmol) was added in several portions to a
solution of the compound from example 35a) (3.00 g, 8.92 mmol) in
1,4-dioxane (20 mL) at room temperature. The reaction was heated to
reflux and stirred for 3.5 h. The reaction was cooled to room
temperature and 1N HCl was added to precipitate the product. The
product was filtered and washed with H.sub.2O and Hexanes to give
the title compound (1.28 g, 49%). LCMS (ES.sup.+) m/z 290.9
(MH.sup.+).
35c) Ethyl
5-hydroxy-6-[3-(methyloxy)phenyl]-3-oxo-2-{[2-(trifluoromethyl)phenyl]met-
hyl}-2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride (103 mg,
2.58 mmol) was added to a solution of the compound from example
35b) (300 mg, 1.03 mmol) in N,N-Dimethylformamide (DMF) (2.5 mL) at
0.degree. C. The reaction was brought to room temperature and
stirred for 45 minutes. The temperature was then reduced to
0.degree. C. and 2-(trifluoromethyl)-benzyl bromide (247 mg, 1.03
mmol) was added. The reaction was brought to room temperature and
stirred for 2.5 hours followed by the addition of 1N HCl. The
solution was diluted with EtOAc and H.sub.2O and the layers
separated. The aqueous layer was backextracted with EtOAc several
times. The combined organic layers were washed with Brine, dried
(MgSO.sub.4), filtered and concentrated. The product was purified
by column chromatography (SiO.sub.2, 35-60% EtOAc/Hexanes) give the
title compound as a white solid (329 mg, 71%).1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 12.71 (br. s., 1H) 7.79 (d, J=7.83 Hz,
1H) 7.65 (t, J=7.45 Hz, 1H) 7.52 (t, J=7.58 Hz, 1H) 7.36 (t, J=7.96
Hz, 1H) 7.15-7.27 (m, 3H) 7.02 (ddd, J=8.27, 2.59, 1.01 Hz, 1H)
5.45 (s, 2H) 4.32 (q, J=7.07 Hz, 2H) 3.76 (s, 3H) 1.29 (t, J=7.20
Hz, 3H).
35d)
N-[(5-Hydroxy-6-[3-(methyloxy)phenyl]-3-oxo-2-{[2-(trifluoromethyl)p-
henyl]methyl}-2,3-dihydro-4-pyridazinyl)carbonyl]glycine. Glycine,
sodium salt (137 mg, 1.41 mmol) was added to a solution of the
compound from example 35c) (316 mg, 0.71 mmol) in 2-methoxyethanol
(2.5 mL) at room temperature. The reaction was heated to reflux and
stirred for 2 h. The reaction was cooled back to room temperature
and H.sub.2O was added. The solution was filtered and 1N HCl was
added to precipitate the product. The solid was filtered and washed
with H.sub.2O and Hexanes to give the title compound (266 mg, 79%).
1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 13.00 (s, 1H) 10.18 (t
J=5.31 Hz, 1H) 7.80 (d, J=7.83 Hz, 1H) 7.64 (t, J=7.45 Hz, 1H) 7.53
(t, J=7.71 Hz, 1H) 7.23-7.45 (m, 4H) 6.99-7.11 (m, 1H) 5.55 (s, 2H)
4.13 (d, J=5.81 Hz, 2H) 3.77 (s, 3H).
Example 36
##STR00047##
N-({2-[(2-Chlorophenyl)methyl]-5-hydroxy-6-[3-(methyloxy)phenyl]-3-oxo-2,3-
-dihydro-4-pyridazinyl}carbonyl)glycine
36a) Ethyl
2-[(2-chlorophenyl)methyl]-5-hydroxy-6-[3-(methyloxy)phenyl]-3-oxo-2,3-di-
hydro-4-pyridazinecarboxylate. Sodium hydride (103 mg, 2.58 mmol)
was added to a solution of the compound from example 35b) (300 mg,
1.03 mmol) in N,N-Dimethylformamide (DMF) (2.5 mL) at 0.degree. C.
The reaction was brought to room temperature and stirred for 45
minutes. The temperature was then reduced to 0.degree. C. and
2-chlorobenzyl bromide (0.13 mL, 1.03 mmol) was added. The reaction
was brought to room temperature and stirred for 2.5 hours followed
by the addition of 1N HCl. The solution was diluted with EtOAc and
H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 40-65% EtOAc/Hexanes) give the title compound as a
white solid (294 mg, 69%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 12.67 (s, 1H) 7.50 (td, J=3.60, 2.15 Hz, 1H) 7.29-7.40 (m, 3H)
7.15-7.27 (m, 3H) 7.02 (ddd, J=8.27, 2.59, 1.01 Hz, 1H) 5.35 (s, 2
H) 4.31 (q, J=7.07 Hz, 2H) 3.76 (s, 3H) 1.29 (t, J=7.07 Hz,
3H).
36b)
N-({2-[(2-Chlorophenyl)methyl]-5-hydroxy-6-[3-(methyloxy)phenyl]-3-o-
xo-2,3-dihydro-4-pyridazinyl}carbonyl)glycine. Glycine, sodium salt
(129 mg, 1.33 mmol) was added to a solution of the compound from
example 36a) (275 mg, 0.66 mmol) in 2-methoxyethanol (2.5 mL) at
room temperature. The reaction was heated to reflux and stirred for
2 h. The reaction was cooled back to room temperature and H.sub.2O
was added. The solution was filtered and 1N HCl was added to
precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes to give the title compound (232 mg, 79%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 13.02 (s, 1H) 10.21 (t,
J=4.29 Hz, 1H) 7.48-7.57 (m, 1H) 7.24-7.42 (m, 6H) 7.02-7.08 (m,
1H) 5.46 (s, 2H) 4.14 (d, J=5.56 Hz, 2H) 3.77 (s, 3H).
Example 37
##STR00048##
N-({2-[(2-Chlorophenyl)methyl]-5-hydroxy-6-[2-(methyloxy)phenyl]-3-oxo-2,3-
-dihydro-4-pyridazinyl}carbonyl)glycine
37a) Ethyl
3-(2-{2-(ethyloxy)-1-[2-(methyloxy)phenyl]-2-oxoethylidene}hydrazino)-3-o-
xopropanoate. A solution of diethyl oxylate (2.7 mL, 20.0 mmol) in
THF (40 mL) and Et.sub.2O (40 mmol) was cooled to -78.degree. C.
2-methoxyphenyl magnesium bromide (1.0 M solution in Et.sub.2O, 24
mL, 24.0 mmol) was dropwise added and the solution stirred under a
nitrogen atmosphere for 2 h at -78.degree. C. The reaction was
brought to 0.degree. C. and quenched with 6N HCl. Additional
Et.sub.2O and H.sub.2O were added and the layers separated. The
aqueous phase was backextracted with Et.sub.2O several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The resulting residue was dissolved in
EtOH (40 mL). Ethyl-3-hydrazino-3-oxopropionate (3.22 g, 22.0 mmol)
and catalytic AcOH (0.2 mL, 3.49 mmol) were added along with a few
spatula tips of MgSO.sub.4. The reaction was heated to reflux and
stirred overnight. The reaction was cooled to room temperature and
filtered. The filtrate was concentrated and azeotroped with Toluene
several times. The product was purified by column chromatography
(SiO.sub.2, 20-45% EtOAc/Hexanes) to give the title compound (3.98
g, 59% over 2 steps). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
10.41-11.75 (m, 1H) 7.31-7.56 (m, 2H) 6.88-7.26 (m, 2H) 3.98-4.37
(m, 4H) 3.23-3.82 (m, 5H) 0.95-1.30 (m, 6H).
37b) Ethyl
5-hydroxy-6-[2-(methyloxy)phenyl]-3-oxo-2,3-dihydro-4-pyridazinecarboxyla-
te. KHMDS (1.78 g, 8.92 mmol) was added in several portions to a
solution of the compound from example 37a) (2.00 g, 5.95 mmol) in
1,4-dioxane (13 mL) at room temperature. The reaction was heated to
reflux and stirred for 1 h. Additional KHMDS (595 mg, 2.98 mmol)
was added and the reaction stirred 3 h. The reaction was cooled to
room temperature and 1N HCl was added to precipitate the product.
The product was purified by column chromatography (SiO.sub.2, 0-4%
MeOH/CH.sub.2Cl.sub.2) then precipitation from
CH.sub.2Cl.sub.2/Hexanes to give the title compound (384 mg, 22%).
1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.94 (s, 1H) 11.96 (s,
1H) 7.40-7.49 (m, 1H) 7.24 (dd, J=7.33, 1.77 Hz, 1H) 7.10 (d,
J=7.83 Hz, 1H) 7.02 (td, J=7.39, 0.88 Hz, 1H) 4.29 (q, J=7.07 Hz,
2H) 3.75 (s, 3H) 1.28 (t, J=7.07 Hz, 3H).
37c) Ethyl
2-[(2-chlorophenyl)methyl]-5-hydroxy-6-[2-(methyloxy)phenyl]-3-oxo-2,3-di-
hydro-4-pyridazinecarboxylate. Sodium hydride (70 mg, 1.75 mmol)
was added to a solution of the compound from example 37b) (203 mg,
0.70 mmol) in N,N-Dimethylformamide (DMF) (2.5 mL) at 0.degree. C.
The reaction was brought to room temperature and stirred for 45
minutes. The temperature was then reduced to 0.degree. C. and
2-chlorobenzyl bromide (0.13 mL, 1.03 mmol) was added. The reaction
was brought to room temperature and stirred for 4 h followed by the
addition of 1N HCl. The solution was diluted with EtOAc and
H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 30-55% EtOAc/Hexanes) give the title compound (79 mg,
27%). LCMS (ES.sup.+) m/z 415.1 (MH.sup.+).
37d)
N-({2-[(2-Chlorophenyl)methyl]-5-hydroxy-6-[2-(methyloxy)phenyl]-3-o-
xo-2,3-dihydro-4-pyridazinyl}carbonyl)glycine. Glycine, sodium salt
(35 mg, 0.36 mmol) was added to a solution of the compound from
example 37c) (75 mg, 0.18 mmol) in 2-methoxyethanol (1.5 mL) at
room temperature. The reaction was heated to reflux and stirred for
1.5 h. The reaction was cooled back to room temperature and
H.sub.2O was added. The solution was filtered and 1N HCl was added
to precipitate the product. The product was filtered then
redissolved in MeOH. The solution was concentrated and Et.sub.2O
added. The product was filtered to give the title compound as an
off-white solid (38 mg, 48%). 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 15.76 (s, 1H) 12.98 (s, 1H) 10.12 (t, J=5.68 Hz, 1H)
7.41-7.56 (m, 2H) 7.30-7.40 (m, 2H) 7.29 (dd, J=7.33, 1.77 Hz, 1H)
7.09-7.22 (m, 2H) 7.04 (td, J=7.45, 0.76 Hz, 1H) 5.42 (s, 2H) 4.11
(d, J=5.81 Hz, 2H) 3.76 (s, 3H).
Example 38
##STR00049##
N-{[2-[(2,6-Dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-d-
ihydro-4-pyridazinyl]carbonyl}glycine
38a) Ethyl
2-[(2,6-dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihy-
dro-4-pyridazinecarboxylate. Sodium hydride (66 mg, 1.66 mmol) was
added to a solution of the compound from example 14a) (150 mg, 0.66
mmol) in N,N-Dimethylformamide (DMF) (2.5 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 30
minutes. The temperature was then reduced to 0.degree. C. and
2,6-dichlorobenzyl bromide (159 mg, 0.66 mmol) was added. The
reaction was brought to room temperature and stirred for 3 h
followed by the addition of 1N HCl. The solution was diluted with
EtOAc and H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 20-50% EtOAc/Hexanes) give the title compound (157 mg,
62%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 7.47-7.55 (m, 2H)
7.36-7.44 (m, 1H) 5.35 (s, 2H) 4.29 (q, J=7.16 Hz, 2H) 3.02 (sept,
J=6.74 Hz, 1H) 1.28 (t, J=7.07 Hz, 3H) 0.89 (d, J=6.82 Hz, 6H).
38b)
N-{[2-[(2,6-Dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-
-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt
(74 mg, 0.76 mmol) was added to a solution of the compound from
example 38a) (146 mg, 0.38 mmol) in 2-methoxyethanol (1.5 mL) at
room temperature. The reaction was heated to reflux and stirred for
2 h. The reaction was cooled back to room temperature and H.sub.2O
was added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and washed with
H.sub.2O and Hexanes to give the title compound (124 mg, 79%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 15.83 (s, 1H) 13.09 (s, 1H)
10.21 (t, J=5.56 Hz, 1H) 7.49-7.57 (m, 2H) 7.36-7.47 (m, 1H) 5.47
(s, 2H) 4.11 (d, J=5.56 Hz, 2H) 3.04 (sept, J=6.78 Hz, 1H) 0.93 (d,
J=6.82 Hz, 6H).
Example 39
##STR00050##
N-{[2-[(4-Fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine
39a) Ethyl
2-[(4-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro--
4-pyridazinecarboxylate. Sodium hydride (66 mg, 1.66 mmol) was
added to a solution of the compound from example 14a) (150 mg, 0.66
mmol) in N,N-Dimethylformamide (DMF) (2.5 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 30
minutes. The temperature was then reduced to 0.degree. C. and
4-fluorobenzyl bromide (0.08 mL, 0.66 mmol) was added. The reaction
was brought to room temperature and stirred for 3 h followed by the
addition of 1N HCl. The solution was diluted with EtOAc and
H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 20-50% EtOAc/Hexanes) give the title compound (106 mg,
48%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.26 (s, 1H) 7.33
(ddd, J=12.00, 5.43, 3.03 Hz, 2H) 7.11-7.22 (m, 2H) 5.12 (s, 2H)
4.26 (q, J=7.24 Hz, 2H) 3.15 (sept, J=6.82 Hz, 1H) 1.25 (t, J=7.07
Hz, 3H) 1.16 (d, J=6.82 Hz, 6H).
39b)
N-{[2-[(4-Fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-
-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt (57
mg, 0.59 mmol) was added to a solution of the compound from example
39a) (98 mg, 0.29 mmol) in 2-methoxyethanol (1.5 mL) at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and washed with
H.sub.2O and Hexanes to give the title compound (86 mg, 82%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 15.87 (s, 1H) 12.98 (s, 1H)
10.18 (t, J=5.18 Hz, 1H) 7.36 (ddd, J=11.87, 5.31, 2.78 Hz, 2H)
7.10-7.24 (m, 2H) 5.23 (s, 2 H) 4.10 (d, J=5.81 Hz, 2H) 3.18 (sept,
J=6.82 Hz, 1H) 1.20 (d, J=6.82 Hz, 6 H).
Example 40
##STR00051##
N-[(5-Hydroxy-6-(1-methylethyl)-3-oxo-2-{[2-(trifluoromethyl)phenyl]methyl-
}-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
40a) Ethyl
5-hydroxy-6-(1-methylethyl)-3-oxo-2-{[2-(trifluoromethyl)phenyl]methyl}-2-
,3-dihydro-4-pyridazinecarboxylate. Sodium hydride (53 mg, 1.33
mmol) was added to a solution of the compound from example 14a)
(120 mg, 0.53 mmol) in N,N-Dimethylformamide (DMF) (2.5 mL) at
0.degree. C. The reaction was brought to room temperature and
stirred for 30 minutes. The temperature was then reduced to
0.degree. C. and 2-(trifluoromethyl)-benzyl bromide (127 mg, 0.53
mmol) was added. The reaction was brought to room temperature and
stirred for 3 h followed by the addition of 1N HCl. The solution
was diluted with EtOAc and H.sub.2O and the layers separated. The
aqueous layer was backextracted with EtOAc several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The product was purified by column
chromatography (SiO.sub.2, 20-50% EtOAc/Hexanes) give the title
compound (144 mg, 71%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
12.40 (s, 1H) 7.77 (d, J=7.58 Hz, 1H) 7.64 (t, J=7.58 Hz, 1H) 7.51
(t, J=7.71 Hz, 1H) 7.10 (d, J=7.58 Hz, 1H) 5.34 (s, 2H) 4.28 (q,
J=7.07 Hz, 2H) 3.16 (sept, J=6.78 Hz, 1H) 1.26 (t, J=7.07 Hz, 3H)
1.10 (d, J=6.82 Hz, 6H).
40b)
N-[(5-Hydroxy-6-(1-methylethyl)-3-oxo-2-{[2-(trifluoromethyl)phenyl]-
methyl}-2,3-dihydro-4-pyridazinyl)carbonyl]glycine. Glycine, sodium
salt (69 mg, 0.71 mmol) was added to a solution of the compound
from example 40a) (137 mg, 0.36 mmol) in 2-methoxyethanol (1.5 mL)
at room temperature. The reaction was heated to reflux and stirred
for 2 h. The reaction was cooled back to room temperature and
H.sub.2O was added. The solution was filtered and 1N HCl was added
to precipitate the product. The product was filtered and washed
with H.sub.2O and Hexanes to give the title compound (136 mg, 91%).
1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 10.19 (t, J=3.92 Hz, 1H)
7.78 (d, J=7.83 Hz, 1H) 7.62 (t, J=7.33 Hz, 1H) 7.51 (t, J=7.58 Hz,
1H) 7.13 (d, J=7.83 Hz, 1H) 5.42 (s, 2H) 4.04 (d, J=5.56 Hz, 2H)
3.19 (sept, J=7.07 Hz, 1H) 1.13 (d, J=6.82 Hz, 6 H).
Example 41
##STR00052##
N-{[2-[(3-Cyanophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydr-
o-4-pyridazinyl]carbonyl}glycine
41a) Ethyl
2-[(3-cyanophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-
-pyridazinecarboxylate. Sodium hydride (53 mg, 1.33 mmol) was added
to a solution of the compound from example 14a) (120 mg, 0.53 mmol)
in N,N-Dimethylformamide (DMF) (2.5 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 30
minutes. The temperature was then reduced to 0.degree. C. and
.alpha.-bromo-m-tolunitrile (104 mg, 0.53 mmol) was added. The
reaction was brought to room temperature and stirred for 3 h
followed by the addition of 1N HCl. The solution was diluted with
EtOAc and H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 30-60% EtOAc/Hexanes) to give the title compound (95
mg, 52%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.35 (s, 1H)
7.78 (dt, J=6.88, 1.74 Hz, 1H) 7.72 (s, 1H) 7.52-7.63 (m, 2H) 5.20
(s, 2H) 4.27 (q, J=7.16 Hz, 2H) 3.16 (sept, J=6.82 Hz, 1H) 1.26 (t,
J=7.07 Hz, 3H) 1.16 (d, J=6.82 Hz, 6H).
41b)
N-{[2-[(3-Cyanophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3--
dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt (54
mg, 0.56 mmol) was added to a solution of the compound from example
41a) (95 mg, 0.28 mmol) in 2-methoxyethanol (1.5 mL) at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and washed with
H.sub.2O and Hexanes to give the title compound (49 mg, 47%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 15.92 (s, 1H) 12.97 (s, 1H)
10.13 (t, J=5.31 Hz, 1H) 7.73-7.82 (m, 2H) 7.52-7.66 (m, 2H) 5.31
(s, 2H) 4.09 (d, J=5.56 Hz, 2H) 3.18 (sept, J=6.95 Hz, 1H) 1.19 (d,
J=6.82 Hz, 6H).
Example 42
##STR00053##
N-({5-Hydroxy-6-(1-methylethyl)-2-[(2-nitrophenyl)methyl]-3-oxo-2,3-dihydr-
o-4-pyridazinyl}carbonyl)glycine
42a) Ethyl
5-hydroxy-6-(1-methylethyl)-2-[(2-nitrophenyl)methyl]-3-oxo-2,3-dihydro-4-
-pyridazinecarboxylate. Sodium hydride (88 mg, 2.21 mmol) was added
to a solution of the compound from example 14a) (200 mg, 0.88 mmol)
in N,N-Dimethylformamide (DMF) (3.5 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 30
minutes. The temperature was then reduced to 0.degree. C. and
2-nitrobenzyl bromide (191 mg, 0.88 mmol) was added. The reaction
was brought to room temperature and stirred for 3 h followed by the
addition of 1N HCl. The solution was diluted with EtOAc and
H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 30-60% EtOAc/Hexanes) to give the title compound (206
mg, 65%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.37 (s, 1H)
8.07 (dd, J=8.21, 1.14 Hz, 1H) 7.72 (td, J=7.58, 1.26 Hz, 1H)
7.54-7.62 (m, 1H) 7.29 (dd, J=7.71, 1.14 Hz, 1H) 5.46 (s, 2H) 4.27
(q, J=7.07 Hz, 2H) 3.13 (sept, J=6.78 Hz, 1H) 1.25 (t, J=7.20 Hz,
3H) 1.09 (d, J=6.82 Hz, 6H).
42b)
N-({5-Hydroxy-6-(1-methylethyl)-2-[(2-nitrophenyl)methyl]-3-oxo-2,3--
dihydro-4-pyridazinyl}carbonyl)glycine. Glycine, sodium salt (107
mg, 1.10 mmol) was added to a solution of the compound from example
42a) (200 mg, 0.55 mmol) in 2-methoxyethanol (1.5 mL) at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and washed with
H.sub.2O and Hexanes to give the title compound (157 mg, 73%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 15.94 (s, 1H) 12.98 (s, 1H)
10.08 (t, J=5.43 Hz, 1H) 8.10 (dd, J=8.08, 1.26 Hz, 1H) 7.72 (td,
J=7.58, 1.26 Hz, 1H) 7.55-7.64 (m, 1H) 7.31 (dd, J=7.58, 1.01 Hz,
1H) 5.57 (s, 2H) 4.10 (d, J=5.56 Hz, 2H) 3.16 (sept, J=6.78 Hz, 1H)
1.13 (d, J=6.82 Hz, 6H).
Example 43
##STR00054##
N-{[2-[(2-Fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine
43a) Ethyl
2-[(2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro--
4-pyridazinecarboxylate. Sodium hydride (53 mg, 1.33 mmol) was
added to a solution of the compound from example 14a) (120 mg, 0.53
mmol) in N,N-Dimethylformamide (DMF) (2.5 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 30
minutes. The temperature was then reduced to 0.degree. C. and
2-fluorobenzyl bromide (0.06 mL, 0.53 mmol) was added. The reaction
was brought to room temperature and stirred for 3 h followed by the
addition of 1N HCl. The solution was diluted with EtOAc and
H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 20-50% EtOAc/Hexanes) give the title compound (110 mg,
62%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.30 (s, 1H)
7.30-7.39 (m, 1H) 7.10-7.26 (m, 3H) 5.20 (s, 2H) 4.27 (q, J=7.07
Hz, 2H) 3.15 (sept, J=6.82 Hz, 1H) 1.26 (t, J=7.07 Hz, 3H) 1.13 (d,
J=6.82 Hz, 6H).
43b)
N-{[2-[(2-Fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-
-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt (64
mg, 0.66 mmol) was added to a solution of the compound from example
43a) (110 mg, 0.33 mmol) in 2-methoxyethanol (1.5 mL) at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and washed with
H.sub.2O and Hexanes to give the title compound (108 mg, 90%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 15.90 (s, 1H) 12.96 (s, 1H)
10.15 (t, J=5.56 Hz, 1H) 7.32-7.41 (m, 1H) 7.13-7.30 (m, 3H) 5.31
(s, 2H) 4.10 (d, J=5.56 Hz, 2H) 3.17 (sept, J=6.86 Hz, 1H) 1.17 (d,
J=6.82 Hz, 6H).
Example 44
##STR00055##
N-{[2-[(2,5-Difluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-d-
ihydro-4-pyridazinyl]carbonyl}glycine
44a) Ethyl
2-[(2,5-difluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihy-
dro-4-pyridazinecarboxylate. Sodium hydride (53 mg, 1.33 mmol) was
added to a solution of the compound from example 14a) (120 mg, 0.53
mmol) in N,N-Dimethylformamide (DMF) (2.5 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 30
minutes. The temperature was then reduced to 0.degree. C. and
2,5-difluorobenzyl bromide (0.07 mL, 0.53 mmol) was added. The
reaction was brought to room temperature and stirred for 3 h
followed by the addition of 1N HCl. The solution was diluted with
EtOAc and H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 15-30% EtOAc/Hexanes) give the title compound (133 mg,
71%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.35 (s, 1H) 7.28
(td, J=9.22, 4.55 Hz, 1H) 7.15-7.25 (m, 1 H) 7.05 (ddd, J=8.84,
5.68, 3.16 Hz, 1H) 5.18 (s, 2H) 4.27 (q, J=7.07 Hz, 2H) 3.15 (sept,
J=6.82 Hz, 1H) 1.26 (t, J=7.07 Hz, 3H) 1.13 (d, J=6.82 Hz, 6H).
44b)
N-{[2-[(2,5-Difluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-
-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt
(69 mg, 0.71 mmol) was added to a solution of the compound from
example 44a) (125 mg, 0.35 mmol) in 2-methoxyethanol (1.5 mL) at
room temperature. The reaction was heated to reflux and stirred for
2 h. The reaction was cooled back to room temperature and H.sub.2O
was added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and washed with
H.sub.2O and Hexanes to give the title compound (112 mg, 84%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 15.92 (s, 1H) 12.97 (s, 1H)
10.12 (t J=5.56 Hz, 1H) 7.29 (td, J=9.22, 4.55 Hz, 1H) 7.17-7.25
(m, 1H) 7.12 (ddd, J=8.91, 5.62, 3.16 Hz, 1H) 5.29 (s, 2H) 4.10 (d,
J=5.81 Hz, 2H) 3.17 (sept, J=6.86 Hz, 1H) 1.16 (d, J=6.82 Hz,
6H).
Example 45
##STR00056##
N-{[2-[(2-Chlorophenyl)methyl]-6-(1,1-dimethylethyl)-5-hydroxy-3-oxo-2,3-d-
ihydro-4-pyridazinyl]carbonyl}glycine
45a) Ethyl 3,3-dimethyl-2-oxobutanoate. DBU (2.1 mL, 14.0 mmol) was
slowly added to a suspension of trimethyl pyruvic acid (60% aqueous
solution, 2.17 g, 10.0 mmol) in MTBE (15 mL). Bromoethane (1.8 mL,
24.0 mmol) was then added. The reaction was heated in the microwave
at 100.degree. C. for 30 minutes. The reaction was cooled and 10%
NaHCO.sub.3 (aq) solution was added. The layers were separated and
the organic layer was washed again with 10% NaHCO.sub.3 (aq). The
aqueous phase was backextraced with Et.sub.2O several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated to give the title compound as a pale
yellow oil (1.44 g, 91%). 1H NMR (400 MHz, CHLOROFORM-d) .delta.
ppm 4.34 (q, J=7.07 Hz, 4 H) 1.38 (t, J=7.20 Hz, 3H) 1.28 (s,
9H).
45b)
Ethyl-2-{[3-(ethyloxy)-3-oxopropanoyl]hydrazono}-3,3-dimethylbutanoa-
te. Ethyl-3-hydrazino-3-oxopropionate (2.38 g, 16.31 mmol) and
catalytic AcOH (0.15 mL, 2.62 mmol) were added to a solution of the
compound from example 45a) (2.15 g. 13.59 mmol) in EtOH (20 mL).
The reaction was heated in the microwave at 150.degree. C. for 30
minutes. The reaction was then cooled to room temperature and
solvent removed under reduced pressure. The product was purified by
column chromatography (SiO.sub.2, 15-30% EtOAc/Hexanes) to give the
title compound (1.66 g, 43%). 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 10.76-11.13 (m, 1H) 4.01-4.38 (m, 4H) 3.39-3.61 (m, 2H)
1.07-1.33 (m, 15H).
45c) Ethyl
6-(1,1-dimethylethyl)-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylate-
. KOtBu (1M solution in t-BuOH, 9.4 mL, 9.38 mmol) was added to the
compound from example 45b) (1.79 g, 6.25 mmol). The reaction was
heated in the microwave at 150.degree. C. for 20 minutes. The
reaction was cooled and H.sub.2O was added followed by 1N HCl to
precipitate the product. The solid was filtered to give the title
compound (1.04 g, 69%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
12.73 (s, 1H) 12.64 (s, 1H) 4.30 (q, J=7.07 Hz, 2H) 1.30 (s, 9H)
1.28 (t, J=7.20 Hz, 3H).
45d) Ethyl
2-[(2-chlorophenyl)methyl]-6-(1,1-dimethylethyl)-5-hydroxy-3-oxo-2,3-dihy-
dro-4-pyridazinecarboxylate. Sodium hydride (42 mg, 1.00 mmol) was
added to a solution of the compound from example 45c) (10 mg, 0.42
mmol) in N,N-Dimethylformamide (DMF) (2 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 30
minutes. The temperature was then reduced to 0.degree. C. and
2-chlorobenzyl bromide (0.05 mL, 0.42 mmol) was added. The reaction
was brought to room temperature and stirred for 3 h followed by the
addition of 1N HCl. The solution was diluted with EtOAc and
H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 20-50% EtOAc/Hexanes) give the title compound (106 mg,
69%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.67 (s, 1H) 7.48
(ddd, J=7.14, 5.75, 3.54 Hz, 1H) 7.28-7.37 (m, 2H) 7.11-7.19 (m,
1H) 5.23 (s, 2H) 4.29 (q, J=7.24 Hz, 2H) 1.23-1.30 (m, 12H).
45e)
N-{[2-[(2-Chlorophenyl)methyl]-6-(1,1-dimethylethyl)-5-hydroxy-3-oxo-
-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt
(49 mg, 0.51 mmol) was added to a solution of the compound from
example 45d) (93 mg, 0.25 mmol) in 2-methoxyethanol (1.5 mL) at
room temperature. The reaction was heated to reflux and stirred for
2 h. The reaction was cooled back to room temperature and H.sub.2O
was added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and washed with
H.sub.2O and Hexanes to give the title compound (79 mg, 81%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 16.43 (s, 1H) 12.99 (s, 1H)
10.26 (t, J=5.56 Hz, 1H) 7.44-7.53 (m, 1H) 7.28-7.39 (m, 2H)
7.18-7.24 (m, 1H) 5.34 (s, 2H) 4.11 (d, J=5.56 Hz, 2H) 1.28 (s,
9H).
Example 46
##STR00057##
N-{[2-[(4-Bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2-
,3-dihydro-4-pyridazinyl]carbonyl}glycine
46a) Ethyl
2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3--
dihydro-4-pyridazinecarboxylate. To a solution of ethyl
5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinecarboxylate
(9.5 g, 42.0 mmol) in N,N-Dimethylformamide (DMF) (250 ml) at
0.degree. C. was added sodium hydride (60% in oil, 2.52 g, 63.0
mmol) in portions. The reaction mixture was stirred at room
temperature for 45 minutes and then cooled back to 0.degree. C. and
4-bromo-2-fluorobenzyl bromide (12.38 g, 46.2 mmol) was added
portionwise. The mixture was stirred at ambient temperature for 2.5
hours then quenched with 1N HCl (10 ml) and diluted with water (30
ml). The aqueous solution was extracted with ethyl acetate
(2.times.100 ml), the organic layers combined and washed with water
(100 ml) and brine (100 ml), dried over Magnesium sulfate, filtered
and solvents removed with rotary evaporation. The crude oil was
purified by flash column chromatography (10-100% ethyl acetate in
hexanes) to provide the crude product (8 g, .about.75% pure, 35%
yield) as a pale yellow solid. The material was purified by reverse
phase HPLC (C18, 75-90% acetonitrile/0.3 M aqueous ammonium
formate) to give the title compound as a white powder. 1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 1.13 (d, J=6.82 Hz, 6H) 1.26 (t,
J=7.20 Hz, 3H) 3.09-3.17 (m, 1H) 4.26 (q, J=7.07 Hz, 2H) 5.16 (s,
2H) 7.21 (t, J=8.08 Hz, 1H) 7.40 (dd, J=8.34, 1.77 Hz, 1H) 7.57
(dd, J=9.73,1.89 Hz, 1H) 12.31 (s, 1H). MS (ES+) m/e 413
[M+H]+.
46b)
N-{[2-[(4-Bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-
-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. To a 500 mL round
bottom was added ethyl
2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1
-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinecarboxylate (9 g, 21.78
mmol) and glycine sodium salt (5.28 g, 54.4 mmol) in
2-methoxyethanol (150 ml) and the mixture was refluxed at
135.degree. C. for 2 hours. The reaction mixture was diluted with
water (50 ml) and acidified with 1N HCl to give a off-white
precipitate that was collected by filtration and washed with water,
hexanes and ether to give
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine (7.20 g, 16.20 mmol,
74.4% yield). The 98% pure material was recrystallized in ethanol
to yield 7.0 g of white crystalline powder. 1H NMR (400 MHz,
DMSO-d.sub.6) d ppm 15.91 (s, 1H), 12.97 (s, 1H), 10.11 (t, J=5.56
Hz, 1H), 7.58 (dd, J=9.60, 2.02 Hz, 1H), 7.40 (dd, J=8.21, 1.64 Hz,
1H), 7.25 (t, J=8.21 Hz, 1 H), 5.26 (s, 2H), 4.10 (d, J=5.56 Hz,
2H), 3.16 (sept, J=6.85, 6.69 Hz, 1H), 1.16 (d, J=6.82 Hz, 6 H). MS
(ES+) m/e 444 [M+H]+.
Example 47
##STR00058##
N-{[2-[(3,4'-Difluoro-4-biphenylyl)methyl]-5-hydroxy-6-(1
-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 ml microwave tube was added
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine (example 46(b), 40 mg,
0.09 mmol), 4-fluorobenzeneboronic acid (15.2 mg, 0.11 mmol),
potassium carbonate (38 mg, 0.272 mmol),
tetrakis(triphenylphosphine)palladium (0) (3 mg, 2.7 .mu.mol),
1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes. The reaction mixture
was diluted with water (5 ml), acidified with 1N HCl (2 ml), and
extracted with ethyl acetate (20 ml). The organic phase was dried
over MgSO4, filtered, and solvents removed under reduced pressure.
The crude residue was purified by HPLC chromatography (ODS silica,
gradient 25-95% acetonitrile/water (0.1% TFA)) to afford the title
compound (23.5 mg, 0.051 mmol, 57% yield) as a white powder. 1H NMR
(400 MHz, DMSO-d.sub.6) d ppm 15.91 (s, 1H), 12.96 (s, 1H), 10.15
(t, J=5.31 Hz, 1H), 7.69-7.80 (m, 2H), 7.55 (dd, J=11.62, 1.77 Hz,
1H), 7.48 (dd, J=8.08, 1.77 Hz, 1H), 7.23-7.38 (m, 3H), 5.34 (s,
2H), 4.10 (d, J=5.56 Hz, 2H), 3.19 (m, 1H), 1.19 (d, J=6.82 Hz,
6H). MS (ES+) m/e 458 [M+H]+.
Example 48
##STR00059##
N-{[2-{[2-Fluoro-4-(4-pyridinyl)phenyl]methyl}-5-hydroxy-6-(1-methylethyl)-
-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 ml microwave tube was added
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine (example 46(b), 40 mg,
0.09 mmol), (4-nitrophenyl)boronic acid (18.1 mg, 0.11 mmol),
potassium carbonate (38 mg, 0.272 mmol),
tetrakis(triphenylphosphine)palladium (0) (3 mg, 2.7 .mu.mol),
1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes. The reaction mixture
was diluted with water (5 ml), acidified with 1N HCl (2 ml), and
extracted with ethyl acetate (20 ml). The organic phase was dried
over MgSO4, filtered, and solvents removed under reduced pressure.
The crude residue was purified by HPLC chromatography (ODS silica,
gradient 15-95% acetonitrile/water (0.1% TFA)) to afford the title
compound (10 mg, 0.023 mmol, 25% yield) as a white powder. 1H NMR
(400 MHz, DMSO-d.sub.6) d ppm 15.93 (s, 1H), 12.99 (s, 1H), 10.13
(t, J=5.43 Hz, 1H), 8.73 (d, J=5.81 Hz, 2H), 7.91 (d, J=5.56 Hz,
2H), 7.81 (dd, J=11.37, 1.52 Hz, 1 H), 7.70 (dd, J=8.08, 1.52 Hz,
1H), 7.43 (t, J=8.08 Hz, 1H), 5.38 (s, 2H), 4.10 (d, J=5.56 Hz, 2
H), 3.19 (m, 1H), 1.19 (d, J=6.82 Hz, 6H). MS (ES+) m/e 441
[M+H]+.
Example 49
##STR00060##
N-[(6-(1,1-Dimethylethyl)-2-{[4-(1,1-dimethylethyl)phenyl]methyl}-5-hydrox-
y-3-oxo-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
49a) Ethyl
6-(1,1-dimethylethyl)-2-{[4-(1,1-dimethylethyl)phenyl]methyl}-5-hydroxy-3-
-oxo-2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride (42 mg,
1.00 mmol) was added to a solution of the compound from example
45c) (100 mg, 0.42 mmol) in N,N-Dimethylformamide (DMF) (2 mL) at
0.degree. C. The reaction was brought to room temperature and
stirred for 30 minutes. The temperature was then reduced to
0.degree. C. and 4-tert-butylbenzyl bromide (0.08 mL, 0.42 mmol)
was added. The reaction was brought to room temperature and stirred
for 3 h followed by the addition of 1N HCl. The solution was
diluted with EtOAc and H.sub.2O and the layers separated. The
aqueous layer was backextracted with EtOAc several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The product was purified by column
chromatography (SiO.sub.2, 20-50% EtOAc/Hexanes) give the title
compound (116 mg, 72%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
12.56 (s, 1H) 7.36 (ddd, J=8.46, 2.15, 2.02 Hz, 2H) 7.23 (d, J=8.59
Hz, 2H) 5.08 (s, 2H) 4.27 (q, J=7.16 Hz, 2H) 1.33 (s, 9H) 1.22-1.29
(m, 12H).
49b) N-[(6-(1,1-Dimethylethyl)-2-{[4-(1,1
-dimethylethyl)phenyl]methyl}-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinyl)c-
arbonyl]glycine. Glycine, sodium salt (54 mg, 0.55 mmol) was added
to a solution of the compound from example 49a) (107 mg, 0.28 mmol)
in 2-methoxyethanol (1.5 mL) at room temperature. The reaction was
heated to reflux and stirred for 2 h. The reaction was cooled back
to room temperature and H.sub.2O was added. The solution was
filtered and 1N HCl was added to precipitate the product. The
product was filtered and washed with H.sub.2O and Hexanes to give
the title compound (68 mg, 59%). 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 16.36 (s, 1H) 12.98 (s, 1H) 10.30 (t, J=5.43 Hz, 1H)
7.37 (ddd, J=8.46, 2.15, 2.02 Hz, 2H) 7.25 (d, J=8.34 Hz, 2H) 5.19
(s, 2H) 4.09 (d, J=5.56 Hz, 2H) 1.35 (s, 9H) 1.25 (s, 9H).
Example 50
##STR00061##
N-{[2-[(3-Fluoro-4-biphenylyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,-
3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 ml microwave tube was added
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine (example 46(b), 40 mg,
0.09 mmol), phenylboronic acid (13.2 mg, 0.11 mmol), potassium
carbonate (38 mg, 0.272 mmol), tetrakis
(triphenylphosphine)palladium (0) (3 mg, 2.7 .mu.mol), 1,4-Dioxane
(1.5 ml) and Water (0.500 ml). The mixture was irradiated at
100.degree. C. for 20 minutes. The reaction mixture was diluted
with water (5 ml), acidified with 1N HCl (2 ml), and extracted with
ethyl acetate (20 ml). The organic phase was dried over MgSO4,
filtered. and solvents removed under reduced pressure. The crude
residue was purified by HPLC chromatography (ODS silica, gradient
15-95% acetonitrile/water (0.1% TFA)) to afford the title compound
(10 mg, 0.023 mmol, 25% yield) as a white powder. 1H NMR (400 MHz,
DMSO-d.sub.6) d ppm 15.91 (s, 1H), 12.97 (br. s., 1H), 10.16 (t,
J=5.56 Hz, 1H), 7.69 (d, J=7.33 Hz, 2H), 7.55 (dd, J=11.62, 1.52
Hz, 1H), 7.51-7.43 (m, 3H), 7.38-7.42 (m, 1H), 7.35 (t, J=7.96 Hz,
1H), 5.34 (s, 2H), 4.10 (d, J=5.81 Hz, 2H), 3.18 (sept, J=6.82 Hz,
1H), 1.19 (d, J=6.82 Hz, 6H). MS (ES+) m/e 440 [M+H]+.
Example 51
##STR00062##
N-{[2-[(3-Fluoro-4'-nitro-4-biphenylyl)methyl]-5-hydroxy-6-(1-methylethyl)-
-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 mL microwave tube was added
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine (example 46(b), 1 g,
2.261 mmol), 4-nitrobenzenboronic acid (0.377 g, 2.261 mmol),
potassium carbonate (0.938 g, 6.78 mmol),
tetrakis(triphenylphosphine)palladium (0) (0.078 g, 0.068 mmol),
1,4-Dioxane (5 ml) and Water (1.667 ml). The mixture was irradiated
at 100.degree. C. for 20 minutes. The reaction mixture was diluted
with water (10 ml) and acidified with 1N HCl to give a off-white
precipitate that was collected by filtration. The mixture of
products was purified by HPLC chromatography (ODS silica, gradient
10-100% acetonitrile/water (0.1% TFA)) to afford the title compound
(450 mg, 0.920 mmol, 40.7% yield). 1H NMR (400 MHz, DMSO-d.sub.6) d
ppm 15.93 (s, 1H), 12.97 (br. s., 1H), 10.14 (t, J=5.68 Hz, 1H),
8.30 (td, J=9.16, 2.65, 2.34 Hz, 2H), 8.01 (td, J=9.16, 2.65, 2.34
Hz, 2H), 7.73 (dd, J=11.37, 1.77 Hz, 1H), 7.63 (dd, J=7.96, 1.89
Hz, 1H), 7.41 (t, J=7.96 Hz, 1H), 5.37 (s, 2H), 4.10 (d, J=5.56 Hz,
2H), 3.19 (sept, J=6.86 Hz, 1H), 1.19 (d, J=6.82 Hz, 6H). MS (ES+)
m/e 485 [M+H]+.
Example 52
##STR00063##
N-{[2-{[3-Fluoro-4'-(trifluoromethyl)-4-biphenylyl]methyl}-5-hydroxy-6-(1--
methylethyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 ml microwave tube was added
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine (example 46(b), 40 mg,
0.09 mmol), (4-trifluoromethyl-phenyl)boronic acid (18.1 mg, 0.11
mmol), potassium carbonate (38 mg, 0.272 mmol), and
tetrakis(triphenylphosphine)palladium (0) (3 mg, 2.7 .mu.mol),
1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes. The reaction mixture
was diluted with water (5 ml), acidified with 1N HCl (2 ml), and
extracted with ethyl acetate (20 ml). The organic phase was dried
over MgSO4, filtered, and solvents removed under reduced pressure.
The crude residue was purified by HPLC chromatography (ODS silica,
gradient 15-95% acetonitrile/water (0.1% TFA)) to afford the title
compound (25 mg, 0.049 mmol, 55% yield) as a white powder. 1H NMR
(400 MHz, DMSO-d.sub.6) d ppm 15.88 (s, 1H), 12.95 (s, 1H), 10.15
(t, J=6.06 Hz, 1H), 7.93 (d, J=8.08 Hz, 2H), 7.83 (d, J=8.59 Hz,
2H), 7.67 (dd, J=11.37, 1.77 Hz, 1H), 7.58 (dd, J=7.96, 1.89 Hz,
1H), 7.39 (t, J=7.96 Hz, 1 H), 5.36 (s, 2 H), 4.09 (d, J=5.56 Hz,
2H), 3.19 (m, 1H), 1.19 (d, J=6.82 Hz, 6H). MS (ES+) m/e 508
[M+H]+.
Example 53
##STR00064##
N-({6-Cyclohexyl-5-hydroxy-2-[(2-methylphenyl)methyl]-3-oxo-2,3-dihydro-4--
pyridazinyl}carbonyl)glycine
53a) Ethyl
3-{2-[1-cyclohexyl-2-(ethyloxy)-2-oxoethylidene]hydrazino}-3-oxopropanoat-
e. A solution of diethyl oxylate (4.1 mL, 30.0 mmol) in THF (50 mL)
was cooled to -78.degree. C. Cyclohexyl magnesium bromide (1.0M
solution in THF, 36 mL, 36.0 mmol) was dropwise added and the
solution stirred under a nitrogen atmosphere for 1.5 h at
-78.degree. C. The reaction was brought to 0.degree. C. and
quenched with 6N HCl. Et.sub.2O and H.sub.2O were added and the
layers separated. The aqueous phase was backextracted with
Et.sub.2O several times. The combined organic layers were washed
with Brine, dried (MgSO.sub.4), filtered and concentrated. The
resulting residue was dissolved in EtOH (20 mL) and partitioned
between two microwave vials. Ethyl-3-hydrazino-3-oxopropionate
(2.41 g, 18.0 mmol) and catalytic AcOH (0.2 mL, 3.50 mmol) were
added to each vial. The reactions were heated in the microwave at
150.degree. C. for 1 h. The reactions were cooled and combined in a
round bottom flask. The solvent was removed under reduced pressure
and the resulting residue was azeotroped with Toluene several
times. The product was purified by column chromatography
(SiO.sub.2, 10-30% EtOAc/Hexanes) to give the title compound (1.93
g, 21% over 2 steps). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
11.44-11.78 (m, 1H) 3.99-4.34 (m, 4H) 3.44-3.67 (m, 2H) 2.52-2.63
(m, 1H) 1.52-1.84 (m, 5H) 1.04-1.37 (m, 11H).
53b) Ethyl
6-cyclohexyl-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylate.
KHMDS (1.24 g, 6.19 mmol) was added in several portions to a
solution of the compound from example 53a) (1.29 g, 4.13 mmol) in
1,4-dioxane at room temperature. The reaction was heated to reflux
and stirred for 3 h. The reaction was cooled to room temperature
and 6N HCl was added to precipitate the product. The solid was
filtered and washed several times with H.sub.2O and Hexanes to give
the title compound (795 mg, 72%). 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 12.64 (s, 1H) 12.29 (s, 1H) 4.28 (q, J=7.07 Hz, 2H)
2.69-2.88 (m, 1H) 1.60-1.88 (m, 5H) 1.29-1.41 (m, 4H) 1.27 (t,
J=7.07 Hz, 3H) 0.97-1.24 (m, 1H).
53c) Ethyl
6-cyclohexyl-5-hydroxy-2-[(2-methylphenyl)methyl]-3-oxo-2,3-dihydro-4-pyr-
idazinecarboxylate. Sodium hydride (38 mg, 0.94 mmol) was added to
a solution of the compound from example 53b) (100 mg, 0.38 mmol) in
N,N-Dimethylformamide (DMF) (1.5 mL) at 0.degree. C. The reaction
was brought to room temperature and stirred for 30 minutes. The
temperature was then reduced to 0.degree. C. and 2-methylbenzyl
bromide (0.05 mL, 0.38 mmol) was added. The reaction was brought to
room temperature and stirred for 3 h followed by the addition of 1N
HCl. The solution was diluted with EtOAc and H.sub.2O and the
layers separated. The aqueous layer was backextracted with EtOAc
several times. The combined organic layers were washed with Brine,
dried (MgSO.sub.4), filtered and concentrated. The product was
purified by column chromatography (SiO.sub.2, 15-30% EtOAc/Hexanes)
to give the title compound (88 mg, 62%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 12.22 (s, 1H) 7.06-7.22 (m, 3H) 7.01 (d,
J=7.07 Hz, 1H) 5.14 (s, 2H) 4.26 (q, J=7.07 Hz, 2H) 2.77-2.91 (m,
1H) 2.37 (s, 3H) 1.55-1.89 (m, 5H) 1.28-1.42 (m, 4H) 1.25 (t,
J=7.20 Hz, 3H) 1.05-1.23 (m, 1H).
53d)
N-({6-Cyclohexyl-5-hydroxy-2-[(2-methylphenyl)methyl]-3-oxo-2,3-dihy-
dro-4-pyridazinyl}carbonyl)glycine. Glycine, sodium salt (45 mg,
0.46 mmol) was added to a solution of the compound from example
53c) (86 mg, 0.23 mmol) in 2-methoxyethanol (1.5 mL) at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and washed with
H.sub.2O and Hexanes to give the title compound (43 mg, 47%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 15.87 (s, 1H) 12.97 (s, 1H)
10.19 (t, J=5.56 Hz, 1H) 7.08-7.25 (m, 3H) 7.02 (d, J=7.07 Hz, 1H)
5.25 (s, 2H) 4.10 (d, J=5.81 Hz, 2H) 2.80-2.94 (m, 1H) 2.38 (s, 3H)
1.63-1.91 (m, 5H) 1.12-1.45 (m, 5H).
Example 54
##STR00065##
N-({6-Cyclohexyl-2-[(2-fluorophenyl)methyl]-5-hydroxy-3-oxo-2,3-dihydro-4--
pyridazinyl}carbonyl)glycine
54a) Ethyl
6-cyclohexyl-2-[(2-fluorophenyl)methyl]-5-hydroxy-3-oxo-2,3-dihydro-4-pyr-
idazinecarboxylate. Sodium hydride (38 mg, 0.94 mmol) was added to
a solution of the compound from example 53b) (100 mg, 0.38 mmol) in
N,N-Dimethylformamide (DMF) (1.5 mL) at 0.degree. C. The reaction
was brought to room temperature and stirred for 30 minutes. The
temperature was then reduced to 0.degree. C. and 2-fluorobenzyl
bromide (0.05 mL, 0.38 mmol) was added. The reaction was brought to
room temperature and stirred for 3 h followed by the addition of 1N
HCl. The solution was diluted with EtOAc and H.sub.2O and the
layers separated. The aqueous layer was backextracted with EtOAc
several times. The combined organic layers were washed with Brine,
dried (MgSO.sub.4), filtered and concentrated. The product was
purified by column chromatography (SiO.sub.2, 15-30% EtOAc/Hexanes)
to give the title compound (64 mg, 45%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 12.27 (s, 1H) 7.31-7.39 (m, 1H) 7.11-7.24
(m, 3H) 5.20 (s, 2H) 4.26 (q, J=7.07 Hz, 2H) 2.78-2.89 (m, 1H)
1.61-1.87 (m, 5H) 1.28-1.40 (m, 4H) 1.25 (t, J=7.20 Hz, 3 H)
1.08-1.22 (m, 1H).
54b)
N-({6-Cyclohexyl-2-[(2-fluorophenyl)methyl]-5-hydroxy-3-oxo-2,3-dihy-
dro-4-pyridazinyl}carbonyl)glycine. Glycine, sodium salt (32 mg,
0.33 mmol) was added to a solution of the compound from example
54a) (61 mg, 0.16 mmol) in 2-methoxyethanol (1.5 mL) at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and washed with
H.sub.2O and Hexanes to give the title compound (30 mg, 46%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 15.90 (s, 1H) 12.96 (s, 1H)
10.14 (t, J=5.68 Hz, 1H) 7.31-7.41 (m, 1H) 7.11-7.29 (m, 3H) 5.30
(s, 2H) 4.09 (d, J=5.56 Hz, 2H) 2.78-2.93 (m, 1H) 1.61-1.91 (m, 5H)
1.08-1.44 (m, 5H).
Example 55
##STR00066##
N-[(6-Cyclohexyl-5-hydroxy-3-oxo-2-{[4-(trifluoromethyl)phenyl]methyl}-2,3-
-dihydro-4-pyridazinyl)carbonyl]glycine
55a) Ethyl
6-cyclohexyl-5-hydroxy-3-oxo-2-{[4-(trifluoromethyl)phenyl]methyl}-2,3-di-
hydro-4-pyridazinecarboxylate. Sodium hydride (38 mg, 0.94 mmol)
was added to a solution of the compound from example 53b) (100 mg,
0.38 mmol) in N,N-Dimethylformamide (DMF) (1.5 mL) at 0.degree. C.
The reaction was brought to room temperature and stirred for 30
minutes. The temperature was then reduced to 0.degree. C. and
4-(trifluoromethyl)-benzyl bromide (90 mg, 0.38 mmol) was added.
The reaction was brought to room temperature and stirred for 3 h
followed by the addition of 1N HCl. The solution was diluted with
EtOAc and H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 15-30% EtOAc/Hexanes) to give the title compound (103
mg, 64%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.30 (s, 1H)
7.72 (d, J=8.08 Hz, 2H) 7.46 (d, J=8.08 Hz, 2 H) 5.23 (s, 2H) 4.26
(q, J=7.07 Hz, 2H) 2.79-2.89 (m, 1H) 1.61-1.90 (m, 5H) 1.29-1.44
(m, 4 H) 1.25 (t, J=7.07 Hz, 3H) 1.12-1.22 (m, 1H).
55b)
N-[(6-Cyclohexyl-5-hydroxy-3-oxo-2-{[4-(trifluoromethyl)phenyl]methy-
l}-2,3-dihydro-4-pyridazinyl)carbonyl]glycine. Glycine, sodium salt
(45 mg, 0.47 mmol) was added to a solution of the compound from
example 55a) (99 mg, 0.23 mmol) in 2-methoxyethanol (1.5 mL) at
room temperature. The reaction was heated to reflux and stirred for
2 h. The reaction was cooled back to room temperature and H.sub.2O
was added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and washed with
H.sub.2O and Hexanes to give the title compound (49 mg, 47%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 10.40 (br. s., 1H) 7.70 (d,
J=8.08 Hz, 2H) 7.46 (d, J=8.08 Hz, 2H) 5.27 (s, 2H) 3.93 (d, J=5.31
Hz, 2H) 2.81-2.97 (m, 1 H) 1.61-1.90 (m, 5H) 1.12-1.45 (m, 5H).
Example 56
##STR00067##
N-({6-cyclohexyl-2-[(3,4-dichlorophenyl)methyl]-5-hydroxy-3-oxo-2,3-dihydr-
o-4-pyridazinyl}carbonyl)glycine
56a) Ethyl
6-cyclohexyl-2-[(3,4-dichlorophenyl)methyl]-5-hydroxy-3-oxo-2,3-dihydro-4-
-pyridazinecarboxylate. Sodium hydride (38 mg, 0.94 mmol) was added
to a solution of the compound from example 53b) (100 mg, 0.38 mmol)
in N,N-Dimethylformamide (DMF) (1.5 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 30
minutes. The temperature was then reduced to 0.degree. C. and
3,4-dichlorobenzyl bromide (0.07 mL, 0.48 mmol) was added. The
reaction was brought to room temperature and stirred for 3 h
followed by the addition of 1N HCl. The solution was diluted with
EtOAc and H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 15-35% EtOAc/Hexanes) to give the title compound (113
mg, 70%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.32 (s, 1H)
7.62 (d, J=8.34 Hz, 1H) 7.54 (d, J=2.02 Hz, 1 H) 7.23 (dd, J=8.34,
2.02 Hz, 1H) 5.14 (s, 2H) 4.26 (q, J=7.07 Hz, 2H) 2.78-2.91 (m, 1H)
1.62-1.87 (m, 5 H) 1.29-1.44 (m, 4H) 1.25 (t, J=7.07 Hz, 3H)
1.13-1.22 (m, 1H).
56b)
N-({6-Cyclohexyl-2-[(3,4-dichlorophenyl)methyl]-5-hydroxy-3-oxo-2,3--
dihydro-4-pyridazinyl}carbonyl)glycine. Glycine, sodium salt (44
mg, 0.45 mmol) was added to a solution of the compound from example
56a) (96 mg, 0.23 mmol) in 2-methoxyethanol (1.5 mL) at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and washed with
H.sub.2O and Hexanes to give the title compound (77 mg, 74%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 15.91 (s, 1H) 12.98 (s, 1H)
10.12 (t, J=5.56 Hz, 1H) 7.62 (d, J=8.08 Hz, 1H) 7.58 (d, J=2.02
Hz, 1H) 7.26 (dd, J=8.34, 2.02 Hz, 1H) 5.25 (s, 2H) 4.09 (d, J=5.81
Hz, 2H) 2.76-2.93 (m, 1H) 1.62-1.93 (m, 5H) 1.10-1.50 (m, 5H).
Example 57
##STR00068##
N-{[2-{[3-Fluoro-4'-(methylthio)-4-biphenylyl]methyl}-5-hydroxy-6-(1-methy-
lethyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 ml microwave tube was added
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine (example 46(b), 75 mg,
0.17 mmol), [4-(methylthio)phenyl]boronic acid (34 mg, 0.20 mmol),
potassium carbonate (70 mg, 0.51 mmol), and
tetrakis(triphenylphosphine)palladium (0) (6 mg, 5 .mu.mol),
1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes. The reaction mixture
was diluted with water (5 ml), acidified with 1N HCl (2 ml), and
extracted with ethyl acetate (20 ml). The organic phase was washed
with brine, dried over MgSO4, filtered, and solvents removed under
reduced pressure. The crude residue was purified by low pressure
reverse phase c18 (ODS silica, gradient 15-95% acetonitrile/water)
to afford the title compound (5 mg, 0.010 mmol, 6% yield) as an off
white powder. 1H NMR (400 MHz, DMSO-d.sub.6) d ppm 15.92 (s, 1H),
12.95 (s, 1H), 10.16 (t, J=5.43 Hz, 1H), 7.65 (d, J=8.34 Hz, 2H),
7.54 (dd, J=11.62, 1.52 Hz, 1H), 7.48 (dd, J=7.96, 1.64 Hz, 1H),
7.30-7.38 (m, 3H), 5.33 (s, 2H), 4.10 (d, J=5.81 Hz, 2 H),
3.31-3.34 (m, 3H), 3.18 (m, 1H), 1.19 (d, J=6.82 Hz, 6H). MS (ES+)
m/e 486 [M+H]+.
Example 58
##STR00069##
N-{[2-{[3-Fluoro-2'-(methyloxy)-4-biphenylyl]methyl}-5-hydroxy-6-(1-methyl-
ethyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 ml microwave tube was added
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine (example 46(b), 75 mg,
0.17 mmol), 2-methoxyphenyl boronic acid (31 mg, 0.20 mmol),
potassium carbonate (70 mg, 0.51 mmol), and
tetrakis(triphenylphosphine)palladium (0) (6 mg, 5 .mu.mol),
1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes. The reaction mixture
was diluted with water (5 ml), acidified with 1N HCl (2 ml), and
extracted with ethyl acetate (20 ml). The organic phase was washed
with brine, dried over MgSO4, filtered, and solvents removed under
reduced pressure. The crude residue was purified HPLC
chromatography (ODS silica, gradient 15-95% acetonitrile/water) to
afford the title compound (25 mg, 0.053 mmol, 31% yield) as a white
powder. 1H NMR (400 MHz, DMSO-d.sub.6) d ppm 15.91 (s, 1H), 12.93
(br. s., 1H), 10.16 (t, J=5.56 Hz, 1H), 7.23-7.41 (m, 5H), 7.12 (d,
J=7.83 Hz, 1H), 7.03 (dt, J=7.39, 0.88 Hz, 1H), 5.33 (s, 2H), 4.10
(d, J=5.81 Hz, 2H), 3.77 (s, 3H), 3.19 (sept, J=6.86 Hz, 1H), 1.20
(d, J=6.82 Hz, 6H). MS (ES+) m/e 470 [M+H]+.
Example 59
##STR00070##
4'-{[5-{[(Carboxymethyl)amino]carbonyl}-4-hydroxy-3-(1-methylethyl)-6-oxo--
1(6H)-pyridazinyl]methyl}-3'-fluoro-4-biphenylcarboxylic acid
To a 5 ml microwave tube was added
N-{[2-[(4-bromo-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine (example 46(b), 75 mg,
0.17 mmol), 4-carboxybenzeneboronic acid (34 mg, 0.20 mmol),
potassium carbonate (70 mg, 0.51 mmol), and
tetrakis(triphenylphosphine)palladium (0) (6 mg, 5 .mu.mol),
1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes. The reaction mixture
was diluted with water (5 ml), acidified with 1N HCl (2 ml), and
extracted with ethyl acetate (20 ml). The organic phase was washed
with brine, dried over MgSO4, filtered, and solvents removed under
reduced pressure. The crude residue was purified by HPLC
chromatography (ODS silica, gradient 15-95% acetonitrile/water
(0.1% TFA)) to afford the title compound (20 mg, 0.041 mmol, 24%
yield) as a white powder. 1H NMR (400 MHz, DMSO-d.sub.6) d ppm
15.93 (s, 1H), 13.03 (br. s., 2H), 10.15 (t, J=5.43 Hz, 1H), 8.01
(d, J=8.34 Hz, 2H), 7.83 (d, J=8.59 Hz, 2H), 7.64 (dd, J=11.37,
1.77 Hz, 1H), 7.57 (dd, J=7.96, 1.64 Hz, 1H), 7.38 (t, J=7.96 Hz,
1H), 5.35 (s, 2H), 4.10 (d, J=5.81 Hz, 2H), 3.19 (sept, J=6.86 Hz,
1H), 1.19 (d, J=6.82 Hz, 6 H). MS (ES+) m/e 484 [M+H]+.
Example 60
##STR00071##
N-{[2-[(2-Bromophenyl)methyl]-6-(1,1-dimethylethyl)-5-hydroxy-3-oxo-2,3-di-
hydro-4-pyridazinyl]carbonyl}glycine
60a) Ethyl
2-[(2-bromophenyl)methyl]-6-(1,1-dimethylethyl)-5-hydroxy-3-oxo-2,3-dihyd-
ro-4-pyridazinecarboxylate. Sodium hydride (37 mg, 0.92 mmol) was
added to a solution of the compound from example 45c) (100 mg, 0.42
mmol) in N,N-Dimethylformamide (DMF) (1.5 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 30
minutes. The temperature was then reduced to 0.degree. C. and
2-bromobenzyl bromide (104 mg, 0.42 mmol) was added. The reaction
was brought to room temperature and stirred for 3 h followed by the
addition of 1N HCl. The solution was diluted with EtOAc and
H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 10-30% EtOAc/Hexanes) to give the title compound (96
mg, 56%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.68 (s, 1H)
7.65 (dd, J=8.08, 1.26 Hz, 1H) 7.36 (td, J=7.52, 1.14 Hz, 1H) 7.25
(td, J=7.64, 1.64 Hz, 1H) 7.11 (dd, J=7.71, 1.64 Hz, 1H) 5.20 (s,
2H) 4.29 (q, J=7.16 Hz, 2 H) 1.25 (s, 9H) 1.23-1.30 (m, 3H).
60b)
N-{[2-[(2-Bromophenyl)methyl]-6-(1,1-dimethylethyl)-5-hydroxy-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt
(44 mg, 0.45 mmol) was added to a solution of the compound from
example 60a) (93 mg, 0.23 mmol) in 2-methoxyethanol (2 mL) at room
temperature. The reaction was heated to reflux and stirred for 2 h.
The reaction was cooled back to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and washed with
H.sub.2O and Hexanes to give the title compound (29 mg, 29%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 16.43 (s, 1H) 12.99 (s, 1H)
10.26 (t, J=5.18 Hz, 1H) 7.67 (dd, J=7.96, 1.14 Hz, 1H) 7.37 (td,
J=7.58, 1.26 Hz, 1H) 7.27 (td, J=7.71, 1.77 Hz, 1H) 7.16 (dd,
J=7.71, 1.64 Hz, 1H) 5.31 (s, 2H) 4.11 (d, J=5.81 Hz, 2H) 1.27 (s,
9H).
Example 61
##STR00072##
N-{[2-[(4-Bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydr-
o-4-pyridazinyl]carbonyl}glycine
To a solution of ethyl
5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinecarboxylate
(example 46a, 1.32 g, 5.83 mmol) in N,N-Dimethylformamide (DMF) (30
ml) at 0.degree. C. was added sodium hydride (0.5 g, 14.58 mmol) in
portions. The reaction mixture was stirred at room temperature for
30 minutes and then cooled back to 0.degree. C. and 4-bromobenzyl
bromide (1.46 g, 5.83 mmol) was added. The mixture was stirred at
ambient temperature for 2 hours then quenched with 1N HCl (10 ml).
The aqueous solution was extracted with ethyl acetate (2.times.50
ml), the organic layers combined, dried over Magnesium sulfate,
filtered and solvents removed with rotary evaporation. The crude
residue was dissolved in 2-methoxyethanol (10 ml), place in a 20 ml
microwave tube and glycine sodium salt (0.75 g, 7.7 mmol) was
added. The mixture was irradiated at 150.degree. C. for 20 minutes,
diluted with water (15 ml) and acidified with 1N HCl to cause a
precipitate. The precipitate was collected by filtration and dried
to give the product as an off white solid (0.750 g, 1.77 mmol,
30.3%) 1H NMR (400 MHz, DMSO-d.sub.6) d ppm 15.89 (s, 1H), 12.97
(s, 1H), 10.16 (t, J=5.31 Hz, 1H), 7.55 (d, J=8.34 Hz, 2H), 7.26
(d, J=8.34 Hz, 2 H), 5.22 (s, 2H), 4.10 (d, J=5.81 Hz, 2H), 3.18
(m, 1H), 1.19 (d, J=6.82 Hz, 6H). MS (ES+) m/e 425 [M+H]+.
Example 62
##STR00073##
N-{[2-[(4'-Fluoro-4-biphenylyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2-
,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 ml microwave tube was added
N-{[2-[(4-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 61, 40 mg, 0.094 mmol),
4-fluorobenzeneboronic acid (14 mg, 0.10 mmol), potassium carbonate
(40 mg, 0.290 mmol), and tetrakis(triphenylphosphine)palladium (0)
(6 mg, 5 .mu.mol) in 1,4-Dioxane (1.5 ml) and Water (0.500 ml). The
mixture was irradiated at 100.degree. C. for 20 minutes, diluted
with water (5 ml), acidified with 1N HCl (2 ml), and extracted with
ethyl acetate (20 ml). The organic phase was washed with brine,
dried over MgSO4, filtered, and solvents removed under reduced
pressure. The crude residue was purified by HPLC chromatography
(ODS silica, gradient 15-95% acetonitrile/water (0.1% TFA)) to
afford the title compound (16.6 mg, 0.038 mmol, 40% yield) as a
white powder. 1H NMR (400 MHz, DMSO-d.sub.6) d ppm 15.88 (s, 1H),
12.99 (br. s., 1H), 10.20 (t, J=5.56 Hz, 1H), 7.65-7.74 (m, 2H),
7.62 (d, J=8.08 Hz, 2H), 7.39 (d, J=8.34 Hz, 2H), 7.23-7.33 (m,
2H), 5.29 (s, 2H), 4.10 (d, J=5.81 Hz, 2H), 3.20 (sept, J=6.78 Hz,
1H), 1.22 (d, J=6.82 Hz, 6H). MS (ES+) m/e 440 [M+H]+.
Example 63
##STR00074##
N-({5-Hydroxy-6-(1-methylethyl)-2-[(4'-nitro-4-biphenylyl)methyl]-3-oxo-2,-
3-dihydro-4-pyridazinyl}carbonyl)glycine
To a 5 ml microwave tube was added
N-{[2-[(4-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 61, 40 mg, 0.094 mmol),
4-nitrobenzeneboronic acid (19 mg, 0.10 mmol), potassium carbonate
(40 mg, 0.290 mmol), and tetrakis(triphenylphosphine)palladium (0)
(6 mg, 5 .mu.mol) in 1,4-Dioxane (1.5 ml) and Water (0.500 ml). The
mixture was irradiated at 100.degree. C. for 20 minutes, diluted
with water (5 ml), acidified with 1N HCl (2 ml), and extracted with
ethyl acetate (20 ml). The organic phase was washed with brine,
dried over MgSO4, filtered, and solvents removed under reduced
pressure. The crude residue was purified by HPLC chromatography
(ODS silica, gradient 15-95% acetonitrile/water (0.1% TFA)) to
afford the title compound (16.8 mg, 0.036 mmol, 38% yield) as a
white powder. 1H NMR (400 MHz, DMSO-d.sub.6) d ppm 15.90 (s, 1H),
12.96 (br. s., 1H), 10.19 (t, J=5.05 Hz, 1H), 8.30 (ddd, J=9.09,
2.53, 2.27 Hz, 2H), 7.95 (ddd, J=9.35, 2.53, 2.27 Hz, 2H), 7.78 (d,
J=8.34 Hz, 2H), 7.45 (d, J=8.34 Hz, 2H), 5.33 (s, 2H), 4.10 (d,
J=5.56 Hz, 2H), 3.20 (qq, J=7.07, 6.87 Hz, 1H), 1.22 (d, J=6.82 Hz,
6H). MS (ES+) m/e 467 [M+H]+.
Example 64
##STR00075##
N-[(5-Hydroxy-6-(1
-methylethyl)-3-oxo-2-{[4'-(trifluoromethyl)-4-biphenylyl]methyl}-2,3-dih-
ydro-4-pyridazinyl)carbonyl]glycine
To a 5 ml microwave tube was added
N-{[2-[(4-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 61, 40 mg, 0.094 mmol),
4-trifluoromethylphenylboronic acid (22 mg, 0.10 mmol), potassium
carbonate (40 mg, 0.290 mmol), and
tetrakis(triphenylphosphine)palladium (0) (6 mg, 5 .mu.mol) in
1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes, diluted with water (5
ml), acidified with 1N HCl (2 ml), and extracted with ethyl acetate
(20 ml). The organic phase was washed with brine, dried over MgSO4,
filtered, and solvents removed under reduced pressure. The crude
residue was purified by HPLC chromatography (ODS silica, gradient
15-95% acetonitrile/water (0.1% TFA)) to afford the title compound
(17.1 mg, 0.035 mmol, 37% yield) as a white powder. 1H NMR (400
MHz, DMSO-d.sub.6) d ppm 15.89 (s, 1H), 12.98 (br. s., 1H), 10.19
(t, J=5.68 Hz, 1H), 7.88 (d, 2H), 7.81 (d, 2H), 7.72 (d, J=8.34 Hz,
2H), 7.43 (d, J=8.34 Hz, 2H), 5.32 (s, 2H), 4.10 (d, J=5.56 Hz,
2H), 3.20 (sept, J=6.78 Hz, 1H), 1.22 (d, J=6.82 Hz, 6H). MS (ES+)
m/e 490 [M+H]+.
Example 65
##STR00076##
N-[(5-Hydroxy-6-(1-methylethyl)-3-oxo-2-{[4-(4-pyridinyl)phenyl]methyl}-2,-
3-dihydro-4-pyridazinyl)carbonyl]glycine
To a 5 ml microwave tube was added
N-{[2-[(4-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 61, 40 mg, 0.094 mmol),
4-pyridinylboronic acid (14 mg, 0.10 mmol), potassium carbonate (40
mg, 0.290 mmol), and tetrakis(triphenylphosphine)palladium (0) (6
mg, 5 .mu.mol) in 1,4-Dioxane (1.5 ml) and Water (0.500 ml). The
mixture was irradiated at 100.degree. C. for 20 minutes, diluted
with water (5 ml), acidified with 1N HCl (2 ml), and extracted with
ethyl acetate (20 ml). The organic phase was washed with brine,
dried over MgSO4, filtered, and solvents removed under reduced
pressure. The crude residue was purified by HPLC chromatography
(ODS silica, gradient 15-95% acetonitrile/water (0.1% TFA)) to
afford the title compound (11.0 mg, 0.026 mmol, 28% yield) as a
white powder. 1H NMR (400 MHz, DMSO-d.sub.6) d ppm 15.91 (s, 1H),
13.01 (s, 1H), 10.17 (t, J=5.56 Hz, 1H), 8.80 (d, J=6.32 Hz, 2H),
8.04 (d, J=6.32 Hz, 2H), 7.91 (d, J=8.34 Hz, 2H), 7.49 (d, J=8.34
Hz, 2H), 5.35 (s, 2H), 4.10 (d, J=5.56 Hz, 2H), 3.20 (m, 1H), 1.22
(d, J=6.82 Hz, 6 H). MS (ES+) m/e 423 [M+H]+.
Example 66
##STR00077##
N-[(5-Hydroxy-6-(1-methylethyl)-2-{[2'-(methyloxy)-4-biphenylyl]methyl}-3--
oxo-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
To a 5 ml microwave tube was added
N-{[2-[(4-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 61, 40 mg, 0.094 mmol),
2-methoxyphenyl boronic acid (17.2 mg, 0.10 mmol), potassium
carbonate (40 mg, 0.290 mmol), and
tetrakis(triphenylphosphine)palladium (0) (6 mg, 5 .mu.mol) in
1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes, diluted with water (5
ml), acidified with 1N HCl (2 ml), and extracted with ethyl acetate
(20 ml). The organic phase was washed with brine, dried over MgSO4,
filtered, and solvents removed under reduced pressure. The crude
residue was purified by HPLC chromatography (ODS silica, gradient
15-95% acetonitrile/water (0.1% TFA)) to afford the title compound
(15.0 mg, 0.033 mmol, 35% yield) as a white powder. 1H NMR (400
MHz, DMSO-d.sub.6) d ppm 15.88 (s, 1H), 12.97 (s, 1H), 10.21 (t,
J=5.94 Hz, 1H), 7.41-7.47 (m, 2H), 7.31-7.37 (m, 3H), 7.26 (dd,
J=7.58, 1.77 Hz, 1H), 7.10 (d, J=7.58 Hz, 1H), 7.01 (dt, J=7.45,
1.01 Hz, 1H), 5.28 (s, 2H), 4.10 (d, J=5.81 Hz, 2H), 3.74 (s, 3H),
3.20 (m, 1H), 1.23 (d, J=6.82 Hz, 6H). MS (ES+) m/e 452 [M+H]+.
Example 67
##STR00078##
4'-{[5-{[(Carboxymethyl)amino]carbonyl}-4-hydroxy-3-(1-methylethyl)-6-oxo--
1(6H)-pyridazinyl]methyl}-4-biphenylcarboxylic acid
To a 5 ml microwave tube was added
N-{[2-[(4-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 61, 40 mg, 0.094 mmol),
4-carboxybenzeneboronic acid (19 mg, 0.10 mmol), potassium
carbonate (40 mg, 0.290 mmol), and
tetrakis(triphenylphosphine)palladium (0) (6 mg, 5 .mu.mol) in
1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes, diluted with water (5
ml), acidified with 1N HCl (2 ml), and extracted with ethyl acetate
(20 ml). The organic phase was washed with brine, dried over MgSO4,
filtered, and solvents removed under reduced pressure. The crude
residue was purified by HPLC chromatography (ODS silica, gradient
15-95% acetonitrile/water (0.1% TFA)) to afford the title compound
(16.0 mg, 0.034 mmol, 36% yield) as a white powder. 1H NMR (400
MHz, DMSO-d.sub.6) d ppm 15.89 (s, 1H), 12.99 (br. s., 1H), 10.19
(t, J=5.68 Hz, 1H), 8.01 (m, 2H), 7.78 (m, 2H), 7.72 (d, J=8.34 Hz,
2H), 7.42 (d, J=8.59 Hz, 2 H), 5.31 (s, 2H), 4.10 (d, J=5.56 Hz,
2H), 3.20 (qq, J=6.85, 6.69 Hz, 1H), 1.22 (d, J=6.82 Hz, 6 H). MS
(ES+) m/e 466 [M+H]+.
Example 68
##STR00079##
N-{[2-(4-Biphenylylmethyl)-6-(1,1-dimethylethyl)-5-hydroxy-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine
68a) Ethyl
2-(4-biphenylylmethyl)-6-(1,1-dimethylethyl)-5-hydroxy-3-oxo-2,3-dihydro--
4-pyridazinecarboxylate. Sodium hydride (34 mg, 0.86 mmol) was
added to a solution of the compound from example 45c) (94 mg, 0.39
mmol) in N,N-Dimethylformamide (DMF) (1.5 mL) at 0.degree. C. The
reaction was brought to room temperature and stirred for 30
minutes. The temperature was then reduced to 0.degree. C. and
4-(bromomethyl)-biphenyl (97 mg, 0.39 mmol) was added. The reaction
was brought to room temperature and stirred for 3 h followed by the
addition of 1N HCl. The solution was diluted with EtOAc and
H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 20-50% EtOAc/Hexanes) to give the title compound (108
mg, 68%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.60 (s, 1H)
7.60-7.68 (m, 4H) 7.31-7.51 (m, 5H) 5.17 (s, 2H) 4.28 (q, J=7.07
Hz, 2H) 1.33 (s, 9H) 1.27 (t, J=7.20 Hz, 3H).
68b) N-{[2-(4-Biphenylylmethyl)-6-(1,1-dimethylethyl)-5-hydroxy-3
-oxo-2,3 -dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium
salt (52 mg, 0.53 mmol) was added to a solution of the compound
from example 68a) (108 mg, 0.27 mmol) in 2-methoxyethanol (2 mL) at
room temperature. The reaction was heated to reflux and stirred for
2 h. The reaction was cooled back to room temperature and H.sub.2O
was added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and washed with
H.sub.2O and Hexanes. The product was purified by recrystallization
from hot AcOH to give the title compound (44 mg, 37%). 1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 16.40 (s, 1H) 12.97 (s, 1H) 10.31
(t, J=5.43 Hz, 1H) 7.59-7.70 (m, 4H) 7.39-7.50 (m, 4H) 7.31-7.39
(m, 1H) 5.28 (s, 2H) 4.11 (d, J=5.56 Hz, 2H) 1.36 (s, 9H).
Example 69
##STR00080##
N-({2-[(2-Chlorophenyl)methyl]-6-[3-(ethyloxy)-5-fluorophenyl]-5-hydroxy-3-
-oxo-2,3-dihydro-4-pyridazinyl}carbonyl)glycine
69a) Ethyl
6-[3-(ethyloxy)-5-fluorophenyl]-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinec-
arboxylate. KHMDS (7.28 g, 36.50 mmol) was added in several
portions to a solution of the compound from example 30a) (5.00 g,
14.60 mmol) in 1,4-dioxane (55 mL) at room temperature. The
reaction was heated to reflux and stirred for 3 h. The reaction was
cooled to room temperature and 1N HCl was added to precipitate the
product. The solid was filtered and washed with H.sub.2O and
Hexanes to give a mixture of the compound from example 30b) and the
title compound. The product was purified by preparative reversed
phase HPLC (80% NH.sub.4OAc (aq) pH 6.8) to give the title compound
(263 mg, 6%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 13.11 (s,
1H) 7.01-7.13 (m, 2H) 6.91 (dt, J=10.93, 2.37 Hz, 1H) 4.31 (q,
J=7.07 Hz, 2H) 4.07 (q, J=7.07 Hz, 2H) 1.34 (t, J=6.95 Hz, 3H) 1.29
(t, J=7.07 Hz, 3H).
69b) Ethyl
2-[(2-chlorophenyl)methyl]-6-[3-(ethyloxy)-5-fluorophenyl]-5-hydroxy-3-ox-
o-2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride (31 mg, 0.78
mmol) was added to a solution of the compound from example 69a)
(100 mg, 0.31 mmol) in N,N-Dimethylformamide (DMF) (1.5 mL) at
0.degree. C. The reaction was brought to room temperature and
stirred for 30 minutes. The temperature was then reduced to
0.degree. C. and 2-chlorobenzyl bromide (0.04 mL, 0.31 mmol) was
added. The reaction was brought to room temperature and stirred for
3 h followed by the addition of 1N HCl. The solution was diluted
with EtOAc and H.sub.2O and the layers separated. The aqueous layer
was backextracted with EtOAc several times. The combined organic
layers were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 25-55% EtOAc/Hexanes) to give the title compound (69
mg, 50%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 7.47-7.54 (m,
1H) 7.28-7.39 (m, 2H) 7.17-7.24 (m, 1H) 7.03-7.10 (m, 2H) 6.91 (dt,
J=11.05, 2.31 Hz, 1H) 5.34 (s, 2H) 4.30 (q, J=7.07 Hz, 2H) 4.04 (q,
J=6.82 Hz, 2H) 1.33 (t, J=6.95 Hz, 3H) 1.28 (t, J=7.20 Hz, 3H).
69c)
N-({2-[(2-Chlorophenyl)methyl]-6-[3-(ethyloxy)-5-fluorophenyl]-5-hyd-
roxy-3-oxo-2,3-dihydro-4-pyridazinyl}carbonyl)glycine. Glycine,
sodium salt (29 mg, 0.30 mmol) was added to a solution of the
compound from example 69b) (66 mg, 0.15 mmol) in 2-methoxyethanol
(1.5 mL) at room temperature. The reaction was heated to reflux and
stirred for 1.5 h. The reaction was cooled back to room temperature
and H.sub.2O was added followed by 1N HCl to precipitate the
product. The product was filtered and washed with H.sub.2O and
Hexanes to give the title compound (61 mg, 86%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 13.02 (s, 1H) 10.19 (t, J=5.18 Hz, 1H)
7.52 (dd, J=7.71, 1.64 Hz, 1H) 7.30-7.40 (m, 2H) 7.24-7.30 (m, 1H)
7.10-7.20 (m, 2H) 6.95 (dt, J=11.05, 2.31 Hz, 1H) 5.46 (s, 2H) 4.14
(d, J=5.56 Hz, 2H) 4.04 (q, J=6.99 Hz, 2H) 1.33 (t, J=6.95 Hz,
3H).
Example 70
##STR00081##
N-{[2-(2-Biphenylylmethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-
-pyridazinyl]carbonyl}glycine
70a) Ethyl
2-(2-biphenylylmethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-py-
ridazinecarboxylate. To a solution of ethyl
5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinecarboxylate
(example 46(a), 125 mg, 0.55 mmol) in N,N-Dimethylformamide (DMF)
(5 ml) at 0.degree. C. was added sodium hydride (55 mg, 0.138 mmol)
in portions. The reaction mixture was stirred at room temperature
for 45 minutes and then cooled back to 0.degree. C. and
2-phenylbenzyl bromide (101 .mu.l, 0.55 mmol) was added. The
mixture was stirred at ambient temperature for 3 hours then
quenched with 1N HCl (3 ml) extracted with ethyl acetate
(2.times.20 ml). The organic layers were combined, dried over
Magnesium sulfate, filtered and solvents removed with rotary
evaporation. The crude oil was purified by flash column
chromatography (10-100% ethyl acetate in hexanes) to provide the
title compound ethyl
2-(2-biphenylylmethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-py-
ridazinecarboxylate (126 mg, 0.32 mmol, 58% yield) as a pale yellow
oil that was used immediately in the next step. MS (ES+) m/e 393
[M+H]+.
70b)
N-{[2-(2-Biphenylylmethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dih-
ydro-4-pyridazinyl]carbonyl}glycine. To a 20 mL microwave tube was
added the product of example 70a) (0.125 g, 0.318 mmol) and Glycine
Sodium Salt (0.046 g, 0.477 mmol) in 2-methoxyethanol (2 ml) and
the mixture was irradiated at 150.degree. C. for 20 minutes. The
reaction mixture was diluted with water (4 ml) and acidified with
1N HCl. The gelatinous aqueous solution was extracted with ethyl
acetate. The organic phase was dried over MgSO4, filtered, and
solvents removed under reduced pressure. The crude residue was
purified by HPLC chromatography (ODS silica, gradient 25-95%
acetonitrile/water (0.1% TFA)) to afford the title compound (84.0
mg, 0.199 mmol, 62% yield) as an off white powder. 1H NMR (400 MHz,
DMSO-d.sub.6) d ppm 15.83 (s, 1H), 12.94 (s, 1 H), 10.09 (t, J=5.68
Hz, 1H), 7.29-7.50 (m, 7H), 7.22-7.28 (m, 1H), 7.10-7.16 (m, 1H),
5.25 (s, 2H), 4.07 (d, J=5.56 Hz, 2H), 3.13 (sept, J=6.78 Hz, 1H),
1.13 (d, J=6.82 Hz, 6H). MS (ES+) m/e 422 [M+H]+.
Example 71
##STR00082##
N-{[2-(3-Biphenylylmethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-
-pyridazinyl]carbonyl}glycine
71a) Ethyl
2-(3-biphenylylmethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-py-
ridazinecarboxylate. To a solution of ethyl
5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinecarboxylate
(example 46(a), 0.125 g, 0.55 mmol) in N,N-Dimethylformamide (DMF)
(5 ml) at 0.degree. C. was added sodium hydride (0.055 g, 0.138
mmol) in portions. The reaction mixture was stirred at room
temperature for 45 minutes and then cooled back to 0.degree. C. and
3-phenylbenzyl bromide (0.137 g, 0.55 mmol) was added. The mixture
was stirred at ambient temperature for 3 hours then quenched with
1N HCl (3 ml) extracted with ethyl acetate (2.times.20 ml). The
organic layers were combined, dried over Magnesium sulfate,
filtered and solvents removed with rotary evaporation. The crude
oil was purified by flash column chromatography (10-100% ethyl
acetate in hexanes) to provide the title compound ethyl
2-(2-biphenylylmethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-py-
ridazinecarboxylate (125 mg, 0.32 mmol, 58% yield) as a pale yellow
solid that was used immediately in the next step. MS (ES+) m/e 393
[M+H]+
71b)
N-{[2-(3-Biphenylylmethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dih-
ydro-4-pyridazinyl]carbonyl}glycine. To a 20 mL microwave tube was
added the product of example 18a (0.125 g, 0.318 mmol) and Glycine
Sodium Salt (0.046 g, 0.477 mmol) in MethoxyEthanol (2 ml) and the
mixture was irradiated at 150.degree. C. for 20 minutes. The
reaction mixture was diluted with water (4 ml), acidified with 1N
HCl, and extracted with ethyl acetate. The organic phase was dried
over MgSO4, filtered, and solvents removed under reduced pressure.
The crude residue was purified by HPLC chromatography (ODS silica,
gradient 25-95% acetonitrile/water (0.1% TFA)) to afford the title
compound (94.0 mg, 0.223 mmol, 70% yield) as an off white powder.
1H NMR (400 MHz, DMSO-d.sub.6) d ppm 15.88 (s, 1H), 12.97 (br. s.,
1H), 10.19 (t, J=5.56 Hz, 1 H), 7.54-7.70 (m, 4H), 7.40-7.51 (m,
3H), 7.37 (tt, J=7.33, 1.26 Hz, 1H), 7.28 (d, J=7.83 Hz, 1H), 5.33
(s, 2H), 4.09 (d, J=5.81 Hz, 2H), 3.19 (sept, J=6.82 Hz, 1H), 1.21
(d, J=6.82 Hz, 6H). MS (ES+) m/e 422 [M+H]+.
Example 72
##STR00083##
N-({6-(1,1-Dimethylethyl)-5-hydroxy-2-[(2-methylphenyl)methyl]-3-oxo-2,3-d-
ihydro-4-pyridazinyl}carbonyl)glycine
72a) Ethyl
6-(1,1-dimethylethyl)-5-hydroxy-2-[(2-methylphenyl)methyl]-3-oxo-2,3-dihy-
dro-4-pyridazinecarboxylate. Sodium hydride (42 mg, 1.05 mmol) was
added to a solution of the compound from example 45c) (100 mg,
0.416 mmol) in N,N-Dimethylformamide (DMF) (1.5 ml) at 0.degree. C.
The reaction was brought to room temperature and stirred for 40
minutes. The temperature was then reduced to 0.degree. C. and
1-(bromomethyl)-2-methylbenzene (0.06 ml, 0.45 mmol) was added. The
reaction was brought to room temperature and stirred for 3 h
followed by addition of 1N HCl. The solution was diluted with
H.sub.2O and EtOAc and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 10-25% EtOAc/Hexanes) to give the title compound (74
mg, 52%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 12.57 (s, 1H)
7.05-7.23 (m, 4H) 5.14 (s, 2H) 4.28 (q, J=7.16 Hz, 2H) 2.36 (s, 3H)
1.23-1.31 (m, 12H).
72b)
N-({6-(1,1-Dimethylethyl)-5-hydroxy-2-[(2-methylphenyl)methyl]-3-oxo-
-2,3-dihydro-4-pyridazinyl}carbonyl)glycine. Glycine, sodium salt
(37 mg, 0.38 mmol) was added to a solution of the compound from
example 72a) (66 mg, 0.19 mmol) in 2-methoxyethanol (1.5 mL) at
room temperature. The reaction was heated to reflux and stirred for
2 h. The reaction was then cooled to room temperature and H.sub.2O
was added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was filtered and purified by
precipitation from CH.sub.2Cl.sub.2/Hexanes to give the title
compound as an off-white solid (54 mg, 75%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 12.99 (s, 1H) 10.31 (t, J=5.43 Hz, 1H)
7.07-7.23 (m, 4H) 5.25 (s, 2H) 4.10 (d, J=5.56 Hz, 2H) 2.37 (s, 3H)
1.31 (s, 9H).
Example 73
##STR00084##
N-[(6-(3,5-Difluorophenyl)-5-hydroxy-3-oxo-2-{[2-(trifluoromethyl)phenyl]m-
ethyl}-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
73a) Ethyl
6-(3,5-difluorophenyl)-5-hydroxy-3-oxo-2-{[2-(trifluoromethyl)phenyl]meth-
yl}-2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride (33.8 mg,
0.844 mmol) was added to a solution of the compound from example
30b) (100 mg, 0.338 mmol) in N,N-Dimethylformamide (DMF) (2 ml) at
0.degree. C. The reaction was brought to room temperature and
stirred for 40 minutes. The temperature was then reduced to
0.degree. C. and 2-(trifluoromethyl)-benzyl bromide (0.05 ml, 0.328
mmol) was added. The reaction was brought to room temperature and
stirred for 3 h followed by the addition of 1N HCl. The reaction
was diluted with H.sub.2O and EtOAc and the layers separated. The
aqueous layer was backextracted with EtOAc several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The product was purified by column
chromatography (SiO.sub.2, 30-70% EtOAc/Hexanes) to give the title
compound (99 mg, 65%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
7.79 (d, J=7.58 Hz, 1H) 7.65 (t, J=7.58 Hz, 1H) 7.53 (t, J=7.58 Hz,
1H) 7.39-7.46 (m, 2H) 7.36 (tt, J=9.25, 2.37 Hz, 1H) 7.23 (d,
J=7.83 Hz, 1 H) 5.45 (s, 2H) 4.30 (q, J=7.07 Hz, 2H) 1.28 (t,
J=7.07 Hz, 3H).
73b)
N-[(6-(3,5-Difluorophenyl)-5-hydroxy-3-oxo-2-{[2-(trifluoromethyl)ph-
enyl]methyl}-2,3-dihydro-4-pyridazinyl)carbonyl]glycine. Glycine,
sodium salt (40 mg, 0.412 mmol) was added to a solution of the
compound from example 73a) (94 mg, 0.207 mmol) in 2-methoxyethanol
(1.5 ml) at room temperature. The reaction was heated to reflux and
stirred for 2 h. The reaction was cooled back to room temperature
and H.sub.2O was added. The solution was filtered and 1N HCl was
added to precipitate the product. The solid was filtered and washed
with H.sub.2O and Hexanes. The product was purified by
recrystallization from hot CH.sub.2Cl.sub.2 to give the title
compound as a white solid (39 mg, 39%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 13.03 (s, 1H) 10.07-10.18 (m, 1H) 7.81
(d, J=7.58 Hz, 1H) 7.64 (t J=7.45 Hz, 1H) 7.47-7.59 (m, 3H) 7.41
(tt, J=9.32, 2.31 Hz, 1 H) 7.29 (d, J=7.83 Hz, 1H) 5.57 (s, 2H)
4.12 (d, J=5.81 Hz, 2H).
Example 74
##STR00085##
N-({6-(3,5-Difluorophenyl)-2-[(3,5-difluorophenyl)methyl]-5-hydroxy-3-oxo--
2,3-dihydro-4-pyridazinyl}carbonyl)glycine
74a) Ethyl
6-(3,5-difluorophenyl)-2-[(3,5-difluorophenyl)methyl]-5-hydroxy-3-oxo-2,3-
-dihydro-4-pyridazinecarboxylate. Sodium hydride (34 mg, 0.850
mmol) was added to a solution of the compound from example 30b)
(100 mg, 0.338 mmol) in N,N-Dimethylformamide (DMF) (2 ml) at
0.degree. C. The reaction was brought to room temperature and
stirred for 40 minutes. The temperature was then reduced to
0.degree. C. and 3,5-difluorobenzyl bromide (0.044 ml, 0.338 mmol)
was added. The reaction was brought to room temperature and stirred
for 3 h followed by addition of 1N HCl. The solution was diluted
with H.sub.2O and EtOAc and the layers separated. The aqueous layer
was backextracted with EtOAc several times. The combined organic
layers were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The crude product was purified by column
chromatography (SiO.sub.2, 30-70% EtOAc/Hexanes) to give the title
compound (98 mg, 69%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
7.42-7.51 (m, 2H) 7.37 (tt, J=9.32, 2.31 Hz, 1H) 7.19 (tt, J=9.47,
2.40 Hz, 1H) 7.01-7.11 (m, J=14.91, 6.57, 2.27 Hz, 2H) 5.28 (s, 2H)
4.29 (q, J=7.07 Hz, 2H) 1.28 (t, J=7.07 Hz, 3H).
74b)
N-({6-(3,5-Difluorophenyl)-2-[(3,5-difluorophenyl)methyl]-5-hydroxy--
3-oxo-2,3-dihydro-4-pyridazinyl}carbonyl)glycine. Glycine, sodium
salt (45 mg, 0.464 mmol) was added to a solution of the compound
from example 74a) (98 mg, 0.232 mmol) in 2-methoxyethanol (1.5 ml)
at room temperature. The reaction was heated to reflux and stirred
for 2 h. The reaction was cooled back to room temperature and
H.sub.2O was added. The solution was filtered and 1N HCl was added
to precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes. The product was purified by recrystallization
from hot CH.sub.2Cl.sub.2 to give the title compound as a white
solid (53 mg, 51%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
13.01 (s, 1H) 10.15 (t, J=5.18 Hz, 1 H) 7.50-7.62 (m, 2H) 7.41 (tt,
J=9.25, 2.37 Hz, 1H) 7.19 (tt, J=9.47, 2.27 Hz, 1 H) 7.05-7.15 (m,
2H) 5.39 (s, 2H) 4.13 (d, J=5.56 Hz, 2H).
Example 75
##STR00086##
N-[(6-(1,1-Dimethylethyl)-5-hydroxy-3-oxo-2-{[2-(trifluoromethyl)phenyl]me-
thyl}-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
75a) Ethyl
6-(1,1-dimethylethyl)-5-hydroxy-3-oxo-2-{[2-(trifluoromethyl)phenyl]methy-
l}-2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride (41.6 mg,
1.041 mmol) was added to a solution of the compound from example
45c) (100 mg, 0.416 mmol) in N,N-Dimethylformamide (DMF) (2 ml) at
0.degree. C. The reaction was brought to room temperature and
stirred for 1 h. The temperature was then reduced to 0.degree. C.
and 2-(trifluoromethyl)-benzyl bromide (99 mg, 0.416 mmol) was
added. The reaction was brought to room temperature and stirred for
3 h followed by addition of 1N HCl. The solution was diluted with
H.sub.2O and EtOAc and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 10-25% EtOAc/Hexanes) to give the title compound as a
colorless oil (129 mg, 78%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 12.72 (s, 1H) 7.77 (d, J=7.83 Hz, 1H) 7.65 (t, J=7.45 Hz, 1 H)
7.52 (t, J=7.58 Hz, 1H) 7.17 (d, J=7.58 Hz, 1H) 5.33 (s, 2H) 4.29
(q, J=7.16 Hz, 2H) 1.20-1.32 (m, 12H).
75b)
N-[(6-(1,1-Dimethylethyl)-5-hydroxy-3-oxo-2-{[2-(trifluoromethyl)phe-
nyl]methyl}-2,3-dihydro-4-pyridazinyl)carbonyl]glycine. Glycine,
sodium salt (61 mg, 0.629 mmol) was added to a solution of the
compound from example 75a) (125 mg, 0.314 mmol) in 2-methoxyethanol
(2 ml) at room temperature. The reaction was heated to reflux and
stirred for 2 h. The reaction was cooled back to room temperature
and H.sub.2O was added. The solution was filtered and 1N HCl was
added to precipitate the product as a tan gum. The gum was filtered
and washed with H.sub.2O and Hexanes. The product was purified by
recrystallization from hot CH.sub.2Cl.sub.2 to give the title
compound as a white solid (103 mg, 77%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 12.98 (s, 1H) 10.23 (t, J=5.31 Hz, 1H)
7.79 (d, J=7.83 Hz, 1H) 7.64 (t, J=7.33 Hz, 1H) 7.53 (t J=7.58 Hz,
1H) 7.21 (d, J=7.83 Hz, 1H) 5.43 (s, 2H) 4.10 (d, J=5.81 Hz, 2H)
1.28 (s, 9 H).
Example 76
##STR00087##
N-{[2-(1,3-Benzothiazol-2-ylmethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3--
dihydro-4-pyridazinyl]carbonyl}glycine
76a) Ethyl
2-(1,3-benzothiazol-2-ylmethyl)-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dih-
ydro-4-pyridazinecarboxylate. Sodium hydride (53 mg, 1.326 mmol)
was added to a solution of the compound from example 14a) (120 mg,
0.530 mmol) in N,N-Dimethylformamide (DMF) (3 ml) at 0.degree. C.
The reaction was brought to room temperature and stirred for 40
minutes. The temperature was then reduced to 0.degree. C. and
2-(bromomethyl)-1,3-benzothiazole (112 mg, 0.491 mmol) was added.
The reaction was brought to room temperature and stirred for 3 h
followed by the addition of 1N HCl. The solution was diluted with
EtOAc and H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 35-60% EtOAc/Hexanes) to give the title compound as an
orange oil (61 mg, 0.163 mmol, 31%). 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 12.48 (s, 1H) 8.05-8.11 (m, 1H) 7.97-8.02 (m, 1H) 7.52
(ddd, J=8.21, 7.20, 1.26 Hz, 1H) 7.44 (td, J=7.58, 1.26 Hz, 1H)
5.59 (s, 2H) 4.27 (q, J=7.07 Hz, 2H) 3.20 (sept, J=6.82 Hz, 1H)
1.26 (t, J=7.20 Hz, 3H) 1.19 (d, J=6.82 Hz, 6H).
76b)
N-{[2-(1,3-Benzothiazol-2-ylmethyl)-5-hydroxy-6-(1-methylethyl)-3-ox-
o-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt
(29 mg, 0.299 mmol) was added to a solution of the compound from
example 76a) (56 mg, 0.150 mmol) in 2-methoxyethanol (2.5 ml) at
room temperature. The reaction was heated to reflux and stirred for
2 h. The reaction was cooled to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl added to precipitate
the product. The solid was filtered and washed with H.sub.2O and
Hexanes. The product was purified by precipitation from
CH.sub.2Cl.sub.2/Hexanes to give the title compound as a pale
yellow solid (41 mg, 68%). 1H NMR (400 MHz, DMSO-d.sub.6) d ppm
1.22 (d, J=6.82 Hz, 6H) 3.14-3.29 (m, 1 H) 4.10 (d, J=5.56 Hz, 2H)
5.70 (s, 2H) 7.41-7.48 (m, 1H) 7.48-7.56 (m, 1H) 7.96-8.04 (m, 1H)
8.06-8.12 (m, 1H) 10.07 (t, J=5.43 Hz, 1H) 12.99 (s, 1H) 16.06 (s,
1H).
Example 77
##STR00088##
N-[(6-(1,1-Dimethylethyl)-2-{[4-fluoro-2-(trifluoromethyl)phenyl]methyl}-5-
-hydroxy-3-oxo-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
77a) Ethyl
6-(1,1-dimethylethyl)-2-{[4-fluoro-2-(trifluoromethyl)phenyl]methyl}-5-hy-
droxy-3-oxo-2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride (46
mg, 1.150 mmol) was added to a solution of the compound from
example 45c) (110 mg, 0.458 mmol) in N,N-Dimethylformamide (DMF)
(2.5 ml) at 0.degree. C. The reaction was brought to room
temperature and stirred for 40 minutes. The temperature was then
reduced to 0.degree. C. and 4-fluoro-2-(trifluoromethyl)-benzyl
bromide (118 mg, 0.458 mmol) was added. The reaction was brought to
room temperature and stirred for 3 h followed by the addition of 1N
HCl. The solution was diluted with EtOAc and H.sub.2O and the
layers separated. The aqueous layer was backextracted with EtOAc
several times. The combined organic layers were washed with Brine,
dried (MgSO.sub.4), filtered and concentrated. The product was
purified by column chromatography (SiO.sub.2, 25-50% EtOAc/Hexanes)
to give the title compound as a colorless oil (136 mg, 71%). 1H NMR
(400 MHz, DMSO-d.sub.6) .delta. ppm 12.71 (s, 1H) 7.68 (dd, J=9.35,
2.78 Hz, 1H) 7.54 (td, J=8.53, 2.65 Hz, 1H) 7.30 (dd, J=8.72, 5.43
Hz, 1H) 5.29 (s, 2H) 4.29 (q, J=7.07 Hz, 2H) 1.27 (t, J=7.07 Hz,
3H) 1.24 (s, 9H).
77b)
N-[(6-(1,1-dimethylethyl)-2-{[4-fluoro-2-(trifluoromethyl)phenyl]met-
hyl}-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazinyl)carbonyl]glycine.
Glycine, sodium salt (27 mg, 0.365 mmol) was added to a solution of
the compound from example 77a) (75 mg, 0.180 mmol) in
2-methoxyethanol (2 ml) at room temperature. The reaction was
heated to reflux and stirred for 2 h. The reaction was then cooled
to room temperature and H.sub.2O added. The solution was filtered
and 1N HCl added to precipitate the product. The solid was filtered
then redissolved in a 1N NaOH solution in MeOH/THF. After stirring
for 1 h at room temperature, the solvent was removed under reduced
pressure and H.sub.2O was added. 1N HCl was added to precipitate
the product, which was filtered as a tan gum. The gum was dissolved
in hot CH.sub.2Cl.sub.2 and the solution concentrated to give the
title compound as an off-white solid (60 mg, 75%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 10.23 (t, J=5.31 Hz, 1H) 7.69 (dd,
J=9.09, 2.78 Hz, 1H) 7.53 (td, J=8.46, 2.78 Hz, 1H) 7.34 (dd,
J=8.59, 5.31 Hz, 1H) 5.39 (s, 2H) 4.09 (d, J=5.81 Hz, 2H) 1.26 (s,
9H).
Example 78
##STR00089##
N-{[2-[(2-Bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydr-
o-4-pyridazinyl]carbonyl}glycine
78a) Ethyl
2-[(2-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-
-pyridazinecarboxylate. To a solution of ethyl
5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinecarboxylate
(example 46(a), 3 g, 13.26 mmol) in N,N-Dimethylformamide (DMF) (50
ml) at 0.degree. C. was added sodium hydride (0.796 g, 19.89 mmol)
in portions. The reaction mixture was stirred at room temperature
for 45 minutes and then cooled back to 0.degree. C. and
2-bromobenzylbromide (3.31 g, 13.26) was added portionwise. The
mixture was stirred at ambient temperature for 2.5 hours then
quenched with 1N HCl (10 ml) and diluted with water (30 ml). The
aqueous solution was extracted with ethyl acetate (2.times.100 ml),
the organic layers combined and washed with water (100 ml) and
brine (100 ml), dried over Magnesium sulfate, filtered and solvents
removed with rotary evaporation. The crude solid was triturated
with ether and filtered. The solid material was 100% starting
material (1 g). The filtrate was purified by flash column
chromatography (10-100% ethyl acetate in hexanes) to provide the
title compound (ethyl
2-[(2-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-
-pyridazinecarboxylate (1.3 g, 2.96 mmol, 22.32% yield) as a pale
yellow solid. 1H NMR (400 MHz, DMSO-d.sub.6) d ppm 12.33 (br. s.,
1H), 7.66 (dd, J=8.08, 1.26 Hz, 1H), 7.35 (td, J=7.52, 1.14 Hz,
1H), 7.25 (td, J=7.58, 1.77 Hz, 1H), 7.03 (dd, J=7.83, 1.52 Hz,
1H), 5.20 (s, 2H), 4.28 (q, J=7.07 Hz, 2H), 3.15 (sept, J=6.82 Hz,
1H), 1.26 (t, J=7.07 Hz, 3H), 1.11 (d, J=6.82 Hz, 6 H). MS (ES+)
m/e 396 [M+H]+.
78b)
N-{[2-[(2-Bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3--
dihydro-4-pyridazinyl]carbonyl}glycine. To a 20 mL microwave tube
was added ethyl
2-[(2-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-
-pyridazinecarboxylate (1.2 g, 3.04 mmol) and Glycine Sodium Salt
(0.589 g, 6.07 mmol) in 2-methoxyethanol (8 ml) and the mixture was
irradiated at 150.degree. C. for 20 minutes. The reaction mixture
was diluted with water (10 ml) and acidified with 1N HCl to give a
off-white precipitate that was collected by filtration and washed
with water, hexanes and ether to give
N-{[2-[(2-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2-
,3-dihydro-4-pyridazinyl]carbonyl}glycine (1.06 g, 2.499 mmol, 82%
yield). 1H NMR (400 MHz, DMSO-d.sub.6) d ppm 15.94 (s, 1H), 12.99
(s, 1H), 10.15 (t, J=5.43 Hz, 1H), 7.67 (dd, J=7.83, 1.26 Hz, 1H),
7.35 (td, J=7.52, 1.14 Hz, 1H), 7.26 (td, J=7.58, 1.77 Hz, 1 H),
7.08 (dd, J=7.71, 1.64 Hz, 1H), 5.31 (s, 2H), 4.10 (d, J=5.81 Hz,
2H), 3.17 (sept, J=6.82 Hz, 1H), 1.14 (d, J=6.82 Hz, 6H). MS (ES+)
m/e 425 [M+H]+.
Example 79
##STR00090##
N-{[2-[(3-Bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydr-
o-4-pyridazinyl]carbonyl}glycine
79a) Ethyl
2-[(3-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-
-pyridazinecarboxylate. To a solution of ethyl
5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinecarboxylate
(example 46(a), 3 g, 13.26 mmol) in N,N-Dimethylformamide (DMF) (50
ml) at 0.degree. C. was added sodium hydride (0.796 g, 19.89 mmol)
in portions. The reaction mixture was stirred at room temperature
for 45 minutes and then cooled back to 0.degree. C. and
3-bromobenzyl bromide (3.31 g, 13.26 mmol) was added portionwise.
The mixture was stirred at ambient temperature for 2.5 hours then
quenched with 1N HCl (10 ml) and diluted with water (30 ml). The
aqueous solution was extracted with ethyl acetate (2.times.100 ml),
the organic layers combined and washed with water (100 ml) and
brine (100 ml), dried over Magnesium sulfate, filtered and solvents
removed with rotary evaporation. The crude oil was purified by
flash column chromatography (10-100% ethyl acetate in hexanes) to
provide the title compound (ethyl
2-[(3-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-
-pyridazinecarboxylate (890 mg, 1.801 mmol, 13.58% yield) as a pale
yellow solid. The product isolated was a 5:1 mixture of the desired
mono alkylated to the bis-alkylated(4-O-benzyl) (LCMS=564). 1H NMR
(400 MHz, DMSO-d.sub.6) d ppm 12.33 (s, 1H), 7.46-7.53 (m, 2H),
7.24-7.35 (m, 2H), 5.14 (s, 2 H), 4.26 (q, J=7.07 Hz, 2H), 3.17
(sept, J=6.78 Hz, 1H), 1.26 (t, J=7.20 Hz, 3H), 1.16 (d, J=6.82 Hz,
6H). MS (ES+) m/e 396 [M+H]+.
79b)
N-{[2-[(3-Bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3--
dihydro-4-pyridazinyl]carbonyl}glycine. To a 20 mL microwave tube
was added ethyl
2-[(3-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-
-pyridazinecarboxylate (700 mg, 1.771 mmol) and Glycine Sodium Salt
(172 mg, 1.771 mmol) in MethoxyEthanol (8 ml) and the mixture was
irradiated at 150.degree. C. for 20 minutes. The reaction mixture
was diluted with water (10 ml) and acidified with 1N HCl to give a
off-white precipitate that was collected by filtration and washed
with water, hexanes and ether to give
N-{[2-[(3-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2-
,3-dihydro-4-pyridazinyl]carbonyl}glycine (270 mg, 0.636 mmol,
35.9% yield). 1H NMR (400 MHz, DMSO-d.sub.6) d ppm 15.91 (s, 1H),
12.98 (s, 1H), 10.15 (t, J=5.43 Hz, 1H), 7.53 (t, J=1.52 Hz, 1H),
7.50 (dt, J=7.58, 1.77 Hz, 1H), 7.32 (t, J=7.71 Hz, 1 H), 7.28
(ddd, J=7.71, 1.39, 1.26 Hz, 1H), 5.25 (s, 2H), 4.10 (d, J=5.56 Hz,
2H), 3.19 (sept, J=6.78 Hz, 1H), 1.20 (d, J=7.07 Hz, 6H). MS (ES+)
m/e 425 [M+H]+.
Example 80
##STR00091##
N-[(6-[3-(Ethyloxy)-5-fluorophenyl]-5-hydroxy-3-oxo-2-{[2-(trifluoromethyl-
)phenyl]methyl}-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
80a) Ethyl
6-[3-(ethyloxy)-5-fluorophenyl]-5-hydroxy-3-oxo-2-{[2-(trifluoromethyl)ph-
enyl]methyl}-2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride
(24 mg, 0.600 mmol) was added to a solution of the compound from
example 69a) (78 mg, 0.242 mmol) in N,N-Dimethylformamide (DMF) (2
ml) at 0.degree. C. The reaction was brought to room temperature
and stirred for 40 minutes. The temperature was then reduced to
0.degree. C. and 2-(trifluoromethyl)-benzyl bromide (58 mg, 0.243
mmol) was added. The reaction was brought to room temperature and
stirred for 3 h followed by addition of 1N HCl. The reaction was
diluted with H.sub.2O and EtOAc and the layers separated. The
aqueous phase was backextracted with EtOAc several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The product was purified by column
chromatography (SiO.sub.2, 25-50% EtOAc/Hexanes) to give the title
compound (104 mg, 89%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
7.79 (d, J=7.58 Hz, 1H) 7.65 (t, J=7.45 Hz, 1H) 7.53 (t, J=7.71 Hz,
1H) 7.22 (d, J=7.83 Hz, 1H) 7.03-7.10 (m, 2H) 6.91 (dt, J=10.86,
2.27 Hz, 1H) 5.44 (s, 2H) 4.30 (q, J=7.16 Hz, 2H) 4.04 (q, J=6.99
Hz, 2H) 1.32 (t, J=6.95 Hz, 3H) 1.28 (t, J=7.07 Hz, 3H).
80b)
N-[(6-[3-(Ethyloxy)-5-fluorophenyl]-5-hydroxy-3-oxo-2-{[2-(trifluoro-
methyl)phenyl]methyl}-2,3-dihydro-4-pyridazinyl)carbonyl]glycine.
Glycine, sodium salt (41 mg, 0.422 mmol) was added to a solution of
the compound from example 80a) (101 mg, 0.210 mmol) in
2-methoxyethanol (2 ml) at room temperature. The reaction was
heated to reflux and stirred for 2 h. The reaction was cooled back
to room temperature and H.sub.2O was added followed by 1N HCl to
precipitate the product. The solid was filtered and washed with
H.sub.2O and Hexanes. The product was purified by recrystallization
from hot EtOH to give the title compound as a white solid (77 mg,
72%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 13.01 (s, 1H)
10.15 (t, J=5.18 Hz, 1H) 7.81 (d, J=7.58 Hz, 1H) 7.64 (t, J=7.45
Hz, 1H) 7.54 (t, J=7.58 Hz, 1H) 7.29 (d, J=7.83 Hz, 1H) 7.12-7.19
(m, 2H) 6.95 (dt, J=10.93, 2.37 Hz, 1H) 5.56 (s, 2H) 4.13 (d,
J=5.81 Hz, 2H) 4.05 (q, J=6.91 Hz, 2H) 1.33 (t, J=6.95 Hz, 3H).
Example 81
##STR00092##
N-{[5-Hydroxy-6-(1
-methylethyl)-2-({4-[6-(4-methyl-1-piperazinyl)-3-pyridinyl]phenyl}methyl-
)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 mL microwave tube was added
N-{[2-[(4-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 61, 31 mg, 0.073 mmol),
1-methyl-4-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridinyl]p-
iperazine (22.16 mg, 0.073 mmol), potassium carbonate (30.3 mg,
0.219 mmol), and tetrakis(triphenylphosphine)palladium (0) (2.53
mg, 2.192 .mu.mol) in 1,4-Dioxane (1.5 ml) and Water (0.500 ml).
The mixture was irradiated at 100.degree. C. for 20 minutes. The
reaction mixture was diluted with water (4 ml) and acidified with
1N HCl (1 ml) then filtered to remove any residue followed by
purification by HPLC chromatography (ODS silica, gradient 10-75%
acetonitrile/water (0.1% TFA)) to afford the title compound
N-{[5-hydroxy-6-(1-methylethyl)-2-({4-[6-(4-methyl-1-piperazinyl)-3-pyrid-
inyl]phenyl}methyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
(23 mg, 0.036 mmol, 49.7% yield) as a white powder tfa salt. 1H NMR
(400 MHz, DMSO-d.sub.6) d ppm 15.87 (s, 1H), 10.19 (t, J=5.68 Hz,
1H), 9.94 (s, 1 H), 8.48 (d, J=2.53 Hz, 1H), 7.94 (dd, J=9.09, 2.53
Hz, 1H), 7.62 (d, J=8.34 Hz, 2H), 7.38 (d, J=8.34 Hz, 2 H), 7.06
(d, J=9.09 Hz, 1H), 5.28 (s, 2H), 4.47 (d, J=13.39 Hz, 2H), 4.10
(d, J=5.56 Hz, 2H), 3.53 (dd, J=10.23, 6.44 Hz, 2H), 2.99-3.26 (m,
5H), 2.85 (s, 3H), 1.22 (d, J=6.82 Hz, 6H). MS (ES+) m/e 521
[M+H]+.
Example 82
##STR00093##
N-({2-[(3,5-Difluorophenyl)methyl]-6-[3-(ethyloxy)-5-fluorophenyl]-5-hydro-
xy-3-oxo-2,3-dihydro-4-pyridazinyl}carbonyl)glycine
82a) Ethyl
2-[(3,5-difluorophenyl)methyl]-6-[3-(ethyloxy)-5-fluorophenyl]-5-hydroxy--
3-oxo-2,3-dihydro-4-pyridazinecarboxylate. Sodium hydride (23 mg,
0.575 mmol) was added to a solution of the compound from example
69a) (75 mg, 0.233 mmol) in N,N-Dimethylformamide (DMF) (2 ml) at
0.degree. C. The reaction was brought to room temperature and
stirred for 40 minutes. The temperature was then reduced to
0.degree. C. and 3,5-difluorobenzylbromide (0.030 ml, 0.232 mmol)
was added. After stirring for 3 h at room temperature, H.sub.2O was
added followed by 1N HCl to precipitate the product. The solid was
filtered and purified by column chromatography (SiO.sub.2, 25-50%
EtOAc/Hexanes) to give the title compound as a white solid (66 mg,
63%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 7.19 (tt, J=9.44,
2.31 Hz, 1H) 7.00-7.13 (m, 4H) 6.93 (dt, J=11.12, 2.27 Hz, 1H) 5.27
(s, 2H) 4.30 (q, J=7.07 Hz, 2H) 4.07 (q, J=6.99 Hz, 2H) 1.34 (t,
J=7.07 Hz, 3H) 1.28 (t, J=7.20 Hz, 3H).
82b)
N-({2-[(3,5-Difluorophenyl)methyl]-6-[3-(ethyloxy)-5-fluorophenyl]-5-
-hydroxy-3-oxo-2,3-dihydro-4-pyridazinyl}carbonyl)glycine. Glycine,
sodium salt (27 mg, 0.278 mmol) was added to a solution of the
compound from example 82a) (63 mg, 0.141 mmol) in 2-methoxyethanol
(1.5 ml) at room temperature. The reaction was heated to reflux and
stirred for 2 h. The reaction was cooled back to room temperature
and H.sub.2O was added followed by 1N HCl to precipitate the
product. The solid was filtered and washed with H.sub.2O and
Hexanes. The product was purified by recrystallization from hot
EtOH to give the title compound as a white solid (15 mg, 22%). 1H
NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 13.01 (s, 1H) 10.16 (t,
J=5.31 Hz, 1H) 7.15-7.24 (m, 3H) 7.05-7.14 (m, 2H) 6.97 (dt,
J=11.05, 2.31 Hz, 1H) 5.39 (s, 2H) 4.13 (d, J=5.81 Hz, 2H) 4.08 (q,
J=7.07 Hz, 2H) 1.34 (t, J=6.95 Hz, 3H).
Example 83
##STR00094##
N-{[5-Hydroxy-6-(1 -methylethyl)-3-oxo-2-({4-[2-(1
-piperazinyl)-4-pyridinyl]phenyl}methyl)-2,3-dihydro-4-pyridazinyl]carbon-
yl}glycine
To a 5 mL microwave tube was added
N-{[2-[(4-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 61, 31 mg, 0.073 mmol),
1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridinyl]piperazine
(21.13 mg, 0.073 mmol), potassium carbonate (30.3 mg, 0.219 mmol),
and tetrakis(triphenylphosphine)palladium (0) (2.53 mg, 2.192
.mu.mol) in 1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture
was irradiated at 100.degree. C. for 20 minutes. The reaction
mixture was diluted with water (10 ml) and acidified with 1N HCl.
The mixture was then extracted with ethyl acetate (2.times.50 mL)
and the organic layers combined, dried over magnesium sulfate,
filtered, and solvents removed by rotary evaporation. The mixture
of products, by tlc, was purified by HPLC chromatography (ODS
silica, gradient 10-100% acetonitrile/water (0.1% TFA)). However,
the product was not there but found in the aqueous extract, 100%
clean, which was evaporated to afford the title compound
N-{[5-hydroxy-6-(1-methylethyl)-3-oxo-2-({4-[2-(1-piperazinyl)-4-pyridiny-
l]phenyl}methyl)-2,3-dihydro-4-pyridazinyl]carbonyl}glycine (26 mg,
0.049 mmol, 66.7% yield) as a tfa salt. 1H NMR (400 MHz,
DMSO-d.sub.6) d ppm 15.89 (s, 1H), 10.18 (t, J=5.81 Hz, 1H), 8.80
(br. s., 2H), 8.21 (d, J=5.56 Hz, 1H), 7.78 (d, J=8.34 Hz, 2H),
7.43 (d, J=8.34 Hz, 2H), 7.23 (s, 1H), 7.09 (dd, J=5.31, 1.01 Hz,
1H), 5.32 (s, 2H), 4.11 (d, J=5.81 Hz, 2H), 3.76-3.87 (m, 4H),
3.13-3.29 (m, 5H), 1.22 (d, J=7.07 Hz, 6H). MS (ES+) m/e 507
[M+H]+.
Example 84
##STR00095##
N-{[2-{[4-(2,6-Difluoro-4-pyridinyl)phenyl]methyl}-5-hydroxy-6-(1-methylet-
hyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 mL microwave tube was added
N-{[2-[(4-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 61, 31 mg, 0.073 mmol),
2,6-difluoropyridine-4-boronic acid (11.61 mg, 0.073 mmol),
potassium carbonate (30.3 mg, 0.219 mmol), and
tetrakis(triphenylphosphine)palladium (0) (2.53 mg, 2.192 .mu.mol)
in 1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes. The reaction mixture
was diluted with water (4 ml), acidified with 1N HCl (1 ml), and
diluted with methanol (2 ml) then filtered to remove any residue
followed by purification by HPLC chromatography (ODS silica,
gradient 10-75% acetonitrile/water (0.1% TFA)) to afford the title
compound
N-{[2-{[4-(2,6-difluoro-4-pyridinyl)phenyl]methyl}-5-hydroxy-6-(1-methyle-
thyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine (15 mg,
0.033 mmol, 44.8% yield) as a white powder. 1H NMR (400 MHz,
DMSO-d.sub.6) d ppm 15.91 (s, 1H), 12.98 (s, 1H), 10.17 (t, J=5.56
Hz, 1H), 7.89 (d, J=8.34 Hz, 2H), 7.57 (s, 2H), 7.45 (d, J=8.34 Hz,
2H), 5.33 (s, 2H), 4.10 (d, J=5.56 Hz, 2H), 3.20 (sept, J=6.82 Hz,
1H), 1.21 (d, J=6.82 Hz, 6H). MS (ES+) m/e 459 [M+H]+.
Example 85
##STR00096##
N-{[5-Hydroxy-6-(1-methylethyl)-2-({4-[2-(methyloxy)-4-pyridinyl]phenyl}me-
thyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 20 mL microwave tube was added
N-{[2-[(4-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 61, 31 mg, 0.073 mmol),
[2-(methyloxy)-4-pyridinyl]boronic acid (11.18 mg, 0.073 mmol),
potassium carbonate (30.3 mg, 0.219 mmol), and
tetrakis(triphenylphosphine)palladium (0) (2.53 mg, 2.192 .mu.mol)
in 1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes. The reaction mixture
was diluted with water (4 ml) and acidified with 1N HCl (1 ml) then
filtered to remove any residue followed by purification by HPLC
chromatography (ODS silica, gradient 10-75% acetonitrile/water
(0.1% TFA)) to afford the title compound
N-{[5-hydroxy-6-(1-methylethyl)-2-({4-[2-(methyloxy)-4-pyridinyl-
]phenyl}methyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
(15 mg, 0.031 mmol, 43.1% yield). 1H NMR (400 MHz, DMSO-d.sub.6) d
ppm 15.89 (s, 1H), 12.96 (br. s., 1H), 10.18 (t, J=5.56 Hz, 1H),
8.22 (d, J=5.31 Hz, 1H), 7.76 (d, J=8.34 Hz, 2H), 7.42 (d, J=8.34
Hz, 2H), 7.30 (dd, J=5.43, 1.39 Hz, 1H), 7.10 (s, 1H), 5.31 (s,
2H), 4.10 (d, J=5.81 Hz, 2H), 3.89 (s, 3H), 3.19 (sept, J=6.78 Hz,
1H), 1.21 (d, J=6.82 Hz, 6H). MS (ES+) m/e 453 [M+H]+.
Example 86
##STR00097##
N-{[2-[(4'-Fluoro-2-biphenylyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2-
,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 ml microwave tube was added
N-{[2-[(2-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 78(b), 75 mg, 0.177
mmol), 4-fluorobenzeneboronic acid (24.74 mg, 0.177 mmol),
potassium carbonate (73.3 mg, 0.530 mmol), and
tetrakis(triphenylphosphine)palladium (0) (6.13 mg, 5.30 .mu.mol)
in 1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes. The reaction mixture
was diluted with water (5 ml), acidified with 1N HCl (2 ml), and
extracted with ethyl acetate (20 ml). The organic phase was dried
over MgSO4, filtered, and solvents removed under reduced pressure.
The crude residue was purified by HPLC chromatography (ODS silica,
gradient 10-75% acetonitrile/water (0.1% TFA)) to afford the title
compound
N-{[2-[(4'-fluoro-2-biphenylyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine (42 mg, 0.095 mmol,
53.5% yield) as a white powder. 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 15.83 (s, 1H), 12.95 (s, 1H), 10.08 (t, J=5.68 Hz, 1
H), 7.44 (ddd, J=11.94, 5.37, 2.91 Hz, 2H), 7.31-7.38 (m, 2H),
7.22-7.30 (m, 3H), 7.11-7.19 (m, 1H), 5.24 (s, 2H), 4.07 (d, J=5.81
Hz, 2H), 3.13 (sept, J=6.82 Hz, 1H), 1.12 (d, J=6.82 Hz, 6H). MS
(ES+) m/e 440 [M+H]+.
Example 87
##STR00098##
N-{[2-[(2,4-Difluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-d-
ihydro-4-pyridazinyl]carbonyl}glycine
87a) Ethyl
2-[(2,4-difluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihy-
dro-4-pyridazinecarboxylate. Sodium hydride (46 mg, 1.150 mmol) was
added to a solution of the compound from example 14a) (105 mg,
0.464 mmol) in N,N-Dimethylformamide (DMF) (3 ml) at 0.degree. C.
The reaction was brought to room temperature and stirred for 40
minutes. The temperature was then reduced to 0.degree. C. and
2,4-difluorobenzylbromide (0.06 ml, 0.468 mmol) was added. The
reaction was brought to room temperature and stirred for 3 h
followed by the addition of 1N HCl. The solution was diluted with
EtOAc and H.sub.2O and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 15-35% EtOAc/Hexanes) to give the title compound as a
white solid (132 mg, 81%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 12.30 (s, 1H) 7.33 (td, J=8.59, 6.82 Hz, 1H) 7.26 (ddd,
J=10.29, 9.54, 2.65 Hz, 1H) 7.03-7.11 (m, J=8.56, 8.56, 2.59, 1.01
Hz, 1H) 5.16 (s, 2 H) 4.26 (q, J=7.16 Hz, 2H) 3.14 (qq, J=6.95,
6.80 Hz, 1H) 1.26 (t, J=7.20 Hz, 3H) 1.12 (d, J=6.82 Hz, 6H).
87b)
N-{[2-[(2,4-Difluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-
-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt
(71 mg, 0.732 mmol) was added to a solution of the compound from
example 87a) (128 mg, 0.363 mmol) in 2-methoxyethanol (2 ml) at
room temperature. The reaction was heated to reflux and stirred for
2 h. The reaction was then cooled to room temperature and H.sub.2O
added. The solution was filtered and 1N HCl was added to
precipitate the product. The product was purified by
recrystallization from hot EtOH to give the title compound as a
white solid (61 mg, 44%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 15.90 (s, 1H) 12.97 (s, 1H) 10.14 (t, J=5.56 Hz, 1H) 7.37 (td,
J=8.53, 6.69 Hz, 1H) 7.27 (ddd, J=10.42, 9.41, 2.65 Hz, 1H)
7.02-7.12 (m, J=8.53, 8.53, 2.53, 0.88 Hz, 1H) 5.27 (s, 2H) 4.09
(d, J=5.81 Hz, 2 H) 3.17 (m, 1H) 1.16 (d, J=7.07 Hz, 6H).
Example 88
##STR00099##
N-{[2-[(3,4-Dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-d-
ihydro-4-pyridazinyl]carbonyl}glycine
88a) Ethyl
2-[(3,4-dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihy-
dro-4-pyridazinecarboxylate. Sodium hydride (46 mg, 1.150 mmol) was
added to a solution of the compound from example 14a) (105 mg,
0.464 mmol) in N,N-Dimethylformamide (DMF) (3 ml) at 0.degree. C.
The reaction was brought to room temperature and stirred for 40
minutes. The temperature was then reduced to 0.degree. C. and
3,4-dichlorobenzyl bromide (0.07 ml, 0.481 mmol) was added. The
reaction was brought to room temperature and stirred for 3 h
followed by the addition of 1N HCl. The reaction was diluted with
H.sub.2O and EtOAc and the layers separated. The aqueous phase was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 15-35% EtOAc/Hexanes) to give the title compound as a
white solid (136 mg, 76%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 12.35 (s, 1H) 7.62 (d, J=8.34 Hz, 1H) 7.54 (d, J=2.02 Hz, 1 H)
7.24 (dd, J=8.34, 2.02 Hz, 1H) 5.14 (s, 2H) 4.26 (q, J=7.07 Hz, 2H)
3.16 (sept, J=6.82 Hz, 1 H) 1.26 (t, J=7.07 Hz, 3H) 1.16 (d, J=6.82
Hz, 6H).
88b)
N-{[2-[(3,4-Dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-
-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt
(68.0 mg, 0.701 mmol) was added to a solution of the compound from
example 88a) (135 mg, 0.350 mmol) in 2-methoxyethanol (2.5 ml) at
room temperature. The reaction was heated to reflux and stirred for
2 h. The reaction was then cooled to room temperature and H.sub.2O
added. The solution was filtered and 1N HCl added to precipitate
the product. The product was filtered then purified by
recrystallization from hot EtOH to give the title compound as a
pale pink solid (81 mg, 56%). 1H NMR (400 MHz, DMSO-d.sub.6)
.delta. ppm 15.92 (s, 1H) 12.98 (s, 1H) 10.13 (t, J=5.31 Hz, 1H)
7.62 (d, J=8.08 Hz, 1H) 7.59 (d, J=1.77 Hz, 1H) 7.27 (dd, J=8.34,
2.02 Hz, 1H) 5.25 (s, 2H) 4.09 (d, J=5.56 Hz, 2H) 3.18 (sept,
J=6.82 Hz, 1H) 1.19 (d, J=6.82 Hz, 6H).
Example 89
##STR00100##
N-({5-Hydroxy-6-(1-methylethyl)-2-[(4'-nitro-2-biphenylyl)methyl]-3-oxo-2,-
3-dihydro-4-pyridazinyl}carbonyl)glycine
To a 5 ml microwave tube was added
N-{[2-[(2-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 78(b), 75 mg, 0.177
mmol), (4-nitrophenyl)boronic acid (29.5 mg, 0.177 mmol), potassium
carbonate (73.3 mg, 0.530 mmol), and
tetrakis(triphenylphosphine)palladium (0) (6.13 mg, 5.30 .mu.mol)
in 1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes. The reaction mixture
was diluted with water (4 ml), acidified with 1N HCl (1 ml), and
diluted with methanol (2 ml) then filtered to remove any residue
followed by purification by HPLC chromatography (ODS silica,
gradient 10-75% acetonitrile/water (0.1% TFA)) to afford the title
compound
N-({5-hydroxy-6-(1-methylethyl)-2-[(4'-nitro-2-biphenylyl)methyl]-3-oxo-2-
,3-dihydro-4-pyridazinyl}carbonyl)glycine (41 mg, 0.087 mmol, 49.2%
yield) as a white powder. 1H NMR (400 MHz, DMSO-d.sub.6) d ppm
10.04 (t, J=5.43 Hz, 1H), 8.27 (ddd, J=9.03, 2.40, 2.21 Hz, 2 H),
7.69 (ddd, J=9.03, 2.40, 2.21 Hz, 2H), 7.37-7.48 (m, 2H), 7.20-7.34
(m, 2H), 5.27 (s, 2H), 4.03 (d, J=5.81 Hz, 2H), 3.10 (sept, J=6.86
Hz, 1H), 1.09 (d, J=6.82 Hz, 6 H). MS (ES+) m/e 467 [M+H]+.
Example 90
##STR00101##
N-[(5-Hydroxy-6-(1-methylethyl)-3-oxo-2-{[4'-(trifluoromethyl)-2-biphenyly-
l]methyl}-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
To a 5 ml microwave tube was added
N-{[2-[(2-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 78(b), 75 mg, 0.177
mmol), 4-trifluoromethylphenylboronic Acid (33.6 mg, 0.177 mmol),
potassium carbonate (73.3 mg, 0.530 mmol), and
tetrakis(triphenylphosphine)palladium (0) (6.13 mg, 5.30 mol) in
1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes. The reaction mixture
was diluted with water (5 ml), acidified with 1N HCl (2 ml), and
extracted with ethyl acetate (20 ml). The organic phase was dried
over MgSO4, filtered, and solvents removed under reduced pressure.
The crude residue was purified by HPLC chromatography (ODS silica,
gradient 10-75% acetonitrile/water (0.1% TFA)) to afford the title
compound N-[(5-hydroxy-6-(1
-methylethyl)-3-oxo-2-{[4'-(trifluoromethyl)-2-biphenylyl]methyl}-2,3-dih-
ydro-4-pyridazinyl)carbonyl]glycine (44 mg, 0.081 mmol, 45.8%
yield) as a white powder. 1H NMR (400 MHz, DMSO-d.sub.6) d ppm
15.81 (s, 1H), 12.99 (s, 1H), 10.05 (t, J=5.56 Hz, 1H), 7.77 (d,
J=8.08 Hz, 2H), 7.61 (d, J=8.08 Hz, 2H), 7.35-7.49 (m, 2H),
7.21-7.35 (m, 2H), 5.27 (s, 2H), 4.06 (d, J=5.56 Hz, 2H), 3.09
(sept, J=6.82 Hz, 1H), 1.09 (d, J=6.82 Hz, 6 H). MS (ES+) m/e 490
[M+H]+.
Example 91
##STR00102##
N-{[2-[(2,3-Dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-d-
ihydro-4-pyridazinyl]carbonyl}glycine
91a) Ethyl
2-[(2,3-dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihy-
dro-4-pyridazinecarboxylate. Sodium hydride (44 mg, 1.100 mmol) was
added to a solution of the compound from example 14a) (100 mg,
0.442 mmol) in N,N-Dimethylformamide (DMF) (3 ml) at 0.degree. C.
The reaction was brought to room temperature and stirred for 40
minutes. The temperature was then reduced to 0.degree. C. and
2,3-dichlorobenzyl bromide (106 mg, 0.442 mmol) was added. The
reaction was brought to room temperature and stirred for 3 h.
H.sub.2O was added followed by 1N HCl to precipitate the product.
The product was filtered and purified by column chromatography
(SiO.sub.2, 15-40% EtOAc/Hexanes) to give the title compound as a
white solid (130 mg, 76%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 12.38 (s, 1H) 7.60 (dd, J=8.08, 1.52 Hz, 1H) 7.35 (t, J=7.83
Hz, 1H) 7.06 (dd, J=7.71, 1.39 Hz, 1H) 5.27 (s, 2H) 4.27 (q, J=7.24
Hz, 2H) 3.15 (sept, J=6.82 Hz, 1H) 1.26 (t, J=7.20 Hz, 3H) 1.11 (d,
J=6.82 Hz, 6H).
91b)
N-{[2-[(2,3-Dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-
-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt
(35 mg, 0.361 mmol) was added to a solution of the compound from
example 91a) (90 mg, 0.234 mmol) in Ethanol (3 ml) at room
temperature. The reaction was heated in the microwave at
150.degree. C. for 20 minutes. The reaction was cooled and H.sub.2O
was added followed by 1N HCl to precipitate the product. The
product was filtered and washed several times with H.sub.2O and
Hexanes to give the title compound as a white solid (72 mg, 74%).
1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 15.96 (s, 1H) 12.98 (s,
1H) 10.11 (t, J=5.43 Hz, 1 H) 7.61 (dd, J=8.08, 1.52 Hz, 1H) 7.34
(t, J=7.83 Hz, 1H) 7.12 (dd, J=7.83, 1.26 Hz, 1H) 5.37 (s, 2H) 4.10
(d, J=5.81 Hz, 2H) 3.17 (m, 1H) 1.14 (d, J=6.82 Hz, 6H).
Example 92
##STR00103##
2'-{[5-{[(carboxymethyl)amino]
carbonyl}-4-hydroxy-3-(1-methylethyl)-6-oxo-1(6H)-pyridazinyl]methyl}-4-b-
iphenylcarboxylic acid
To a 5 ml microwave tube was added
N-{[2-[(2-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 78(b), 75 mg, 0.177
mmol), 4-carboxybenzeneboronic acid (29.3 mg, 0.177 mmol),
potassium carbonate (73.3 mg, 0.530 mmol), and
tetrakis(triphenylphosphine)palladium (0) (6.13 mg, 5.30 .mu.mol)
in 1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes. The reaction mixture
was diluted with water (5 ml), acidified with 1N HCl (2 ml), and
extracted with ethyl acetate (20 ml). The organic phase was dried
over MgSO4, filtered, and solvents removed under reduced pressure.
The crude residue was purified by HPLC chromatography (ODS silica,
gradient 10-75% acetonitrile/water (0.1% TFA)) to afford the title
compound
2'-{[5-{[(carboxymethyl)amino]carbonyl}-4-hydroxy-3-(1-methylethyl)-6-oxo-
-1(6H)-pyridazinyl]methyl}-4-biphenylcarboxylic acid (non-preferred
name) (30 mg, 0.064 mmol, 36.1% yield) as a white powder. 1H NMR
(400 MHz, DMSO-d.sub.6) d ppm 15.82 (s, 1H), 12.99 (br. s., 2 H),
10.06 (t, J=5.68 Hz, 1H), 7.99 (d, J=8.34 Hz, 2H), 7.52 (d, J=8.34
Hz, 2 H), 7.38 (dt, J=4.36, 2.24 Hz, 2H), 7.25-7.32 (m, 1H),
7.17-7.24 (m, 1H), 5.26 (s, 2H), 4.06 (d, J=5.56 Hz, 2H), 3.11
(sept, J=6.82 Hz, 1H), 1.10 (d, J=6.82 Hz, 6H). MS (ES+) m/e 466
[M+H]+.
Example 93
##STR00104##
N-[(5-Hydroxy-6-(1-methylethyl)-2-{[2'-(methyloxy)-2-biphenylyl]methyl}-3--
oxo-2,3-dihydro-4-pyridazinyl)carbonyl]glycine
To a 5 ml microwave tube was added
N-{[2-[(2-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 78(b), 75 mg, 0.177
mmol), 2-methoxyphenylboronic acid (26.9 mg, 0.177 mmol), potassium
carbonate (73.3 mg, 0.530 mmol), and
tetrakis(triphenylphosphine)palladium (0) (6.13 mg, 5.30 .mu.mol)
in 1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture was
irradiated at 100.degree. C. for 20 minutes. The reaction mixture
was diluted with water (5 ml), acidified with 1N HCl (2 ml), and
extracted with ethyl acetate (20 ml). The organic phase was dried
over MgSO4, filtered, and solvents removed under reduced pressure.
The crude residue was purified by HPLC chromatography (ODS silica,
gradient 10-75% acetonitrile/water (0.1% TFA)) to afford the title
compound
N-[(5-hydroxy-6-(1-methylethyl)-2-{[2'-(methyloxy)-2-biphenylyl]methyl}-3-
-oxo-2,3-dihydro-4-pyridazinyl)carbonyl]glycine (40 mg, 0.088 mmol,
49.6% yield) as a white powder. 1H NMR (400 MHz, DMSO-d.sub.6) d
ppm 15.80 (s, 1H), 12.99 (br. s., 1H), 10.09 (t, J=4.93 Hz, 1 H),
7.26-7.39 (m, 3H), 7.04-7.19 (m, 4H), 6.98 (td, J=7.45, 0.76 Hz,
1H), 5.00-5.16 (m, rotomers, 2H), 4.06 (d, J=5.81 Hz, 2H), 3.74 (s,
3H), 3.08 (m, 1H), 1.03-1.13 (m, rotomers, 6H). MS (ES+) m/e 452
[M+H]+.
Example 94
##STR00105##
N-{[2-[(4-Chloro-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo--
2,3-dihydro-4-pyridazinyl]carbonyl}glycine
94a) Ethyl
2-[(4-chloro-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-
-dihydro-4-pyridazinecarboxylate. Sodium hydride (44 mg, 1.100
mmol) was added to a solution of the compound from example 14a)
(100 mg, 0.442 mmol) in N,N-Dimethylformamide (DMF) (3 ml) at
0.degree. C. The reaction was brought to room temperature and
stirred for 40 minutes. The temperature was then reduced to
0.degree. C. and 4-chloro-2-fluorobenzyl bromide (99 mg, 0.442
mmol) was added. The reaction was brought to room temperature and
stirred for 3 h followed by the addition of 1N HCl. H.sub.2O and
EtOAc were added and the layers separated. The aqueous layer was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 15-35% EtOAc/Hexanes) to give the title compound as a
white solid (124 mg, 76%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 12.32 (s, 1H) 7.45 (dd, J=9.73, 1.14 Hz, 1H) 7.24-7.31 (m, 2 H)
5.17 (s, 2H) 4.26 (q, J=7.24 Hz, 2H) 3.14 (sept, J=6.82 Hz, 1H)
1.26 (t, J=7.07 Hz, 3H) 1.12 (d, J=6.82 Hz, 6H).
94b)
N-{[2-[(4-Chloro-2-fluorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)--
3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium
salt (49 mg, 0.505 mmol) was added to a solution of the compound
from example 94a) (94 mg, 0.255 mmol) in Ethanol (3 ml) at room
temperature. The reaction was heated in the microwave at
150.degree. C. for 20 minutes. The reaction was cooled and H.sub.2O
added. The solution was filtered and 1N HCl added to precipitate
the product. The product was filtered and purified by
recrystallization from hot EtOH to give the title compound as a
white solid (48 mg, 47%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 15.92 (s, 1H) 12.97 (s, 1H) 10.12 (t, J=5.56 Hz, 1H) 7.47 (dd,
J=9.98, 1.89 Hz, 1H) 7.22-7.36 (m, 2H) 5.28 (s, 2H) 4.10 (d, J=5.56
Hz, 2H) 3.16 (qq, J=6.85, 6.69 Hz, 1H) 1.16 (d, J=6.82 Hz, 6H).
Example 95
##STR00106##
N-{[2-[(2,5-Dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-d-
ihydro-4-pyridazinyl]carbonyl}glycine
95a) Ethyl
2-[(2,5-dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihy-
dro-4-pyridazinecarboxylate. Sodium hydride (44 mg, 1.100 mmol) was
added to a solution of the compound from example 14a) (100 mg,
0.442 mmol) in N,N-Dimethylformamide (DMF) (3 ml) at 0.degree. C.
The reaction was brought to room temperature and stirred for 40
minutes. The temperature was then reduced to 0.degree. C. and
2,5-dichlorobenzyl bromide (106 mg, 0.442 mmol) was added. The
reaction was brought to room temperature and stirred for 3 h
followed by the addition of 1N HCl. The reaction was diluted with
H.sub.2O and EtOAc and the layers separated. The aqueous phase was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 15-30% EtOAc/Hexanes) to give the title compound as a
white solid (122 mg, 0.317 mmol, 72%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 12.40 (s, 1H) 7.51-7.55 (m, 1H) 7.43 (dd,
J=8.59, 2.53 Hz, 1H) 7.22 (d, J=2.53 Hz, 1H) 5.22 (s, 2H) 4.28 (q,
J=7.07 Hz, 2H) 3.15 (qq, J=7.07, 6.82 Hz, 1H) 1.27 (t, J=7.07 Hz,
3H) 1.11 (d, J=6.82 Hz, 6H).
95b)
N-{[2-[(2,5-Dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-
-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt
(41 mg, 0.422 mmol) was added to a solution of the compound from
example 95a) (82 mg, 0.213 mmol) in Ethanol (2 ml) at room
temperature. The reaction was heated in the microwave at
150.degree. C. for 15 minutes. The reaction was cooled and H.sub.2O
added. The solution was filtered and 1N HCl added to precipitate
the product. The product was filtered and purified by
recrystallization from hot EtOH to give the title compound as a
white solid (17 mg, 19%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 15.95 (s, 1H) 12.97 (s, 1H) 10.11 (t, J=5.56 Hz, 1H) 7.54 (d,
J=8.59 Hz, 1H) 7.44 (dd, J=8.59, 2.53 Hz, 1H) 7.32 (d, J=2.53 Hz,
1H) 5.32 (s, 2H) 4.10 (d, J=5.56 Hz, 2H) 3.17 (sept, J=6.82 Hz, 1H)
1.14 (d, J=6.82 Hz, 6H).
Example 96
##STR00107##
N-{[2-[(2,4-Dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-d-
ihydro-4-pyridazinyl]carbonyl}glycine
96a) Ethyl
2-[(2,4-dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihy-
dro-4-pyridazinecarboxylate. Sodium hydride (50 mg, 1.250 mmol) was
added to a solution of the compound from example 14a) (113 mg,
0.499 mmol) in N,N-Dimethylformamide (DMF) (3 ml) at 0.degree. C.
The reaction was brought to room temperature and stirred for 40
minutes. The temperature was then reduced to 0.degree. C. and
2,4-dichlorobenzyl chloride (0.07 ml, 0.501 mmol) was added. The
reaction was brought to room temperature and stirred for 3 h
followed by the addition of 1N HCl. The reaction was diluted with
H.sub.2O and EtOAc and the layers separated. The aqueous phase was
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 15-30% EtOAc/Hexanes) to give the title compound as a
white solid (90 mg, 47%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 12.36 (s, 1H) 7.66 (d, J=2.02 Hz, 1H) 7.42 (dd, J=8.34, 2.27
Hz, 1H) 7.16 (d, J=8.34 Hz, 1H) 5.21 (s, 2H) 4.27 (q, J=7.24 Hz,
2H) 3.14 (sept, J=6.78 Hz, 1 H) 1.26 (t, J=7.07 Hz, 3H) 1.11 (d,
J=6.82 Hz, 6H).
96b)
N-{[2-[(2,4-Dichlorophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-
-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. Glycine, sodium salt
(44 mg, 0.453 mmol) was added to a solution of the compound from
example 96a) (88 mg, 0.228 mmol) in 2-methoxyethanol (2 ml) at room
temperature. The reaction was heated to refluxed for 2 h. The
reaction was then cooled to room temperature and H.sub.2O added.
The solution was filtered and 1N HCl added to precipitate the
product. The product was filtered and washed several times with
H.sub.2O and Hexanes to give the title compound as an off-white
solid (43 mg, 45%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
15.94 (s, 1H) 12.98 (s, 1H) 10.12 (t, J=5.56 Hz, 1H) 7.68 (d,
J=2.02 Hz, 1H) 7.41 (dd, J=8.34, 2.27 Hz, 1H) 7.21 (d, J=8.34 Hz,
1H) 5.32 (s, 2H) 4.10 (d, J=5.81 Hz, 2H) 3.16 (sept, J=6.78 Hz, 1
H) 1.14 (d, J=6.82 Hz, 6H).
Example 97
##STR00108##
N-{[2-[(4'-Amino-2-biphenylyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,-
3-dihydro-4-pyridazinyl]carbonyl}glycine
To a Parr shaker flask was added
N-({5-hydroxy-6-(1-methylethyl)-2-[(4'-nitro-2-biphenylyl)methyl]-3-oxo-2-
,3-dihydro-4-pyridazinyl}carbonyl)glycine (example 89, 65 mg, 0.139
mmol) and palladium on carbon (7.41 mg, 0.070 mmol) in Methanol (10
ml) and Ethyl acetate (2.500 ml) under a nitrogen atmosphere. The
mixture was placed on a Parr shaker under 50 psi Hydrogen for 1
hour. The reaction mixture was filtered to remove the palladium on
carbon followed by removal of the solvent by rotary evaporation and
purification by HPLC chromatography (ODS silica, gradient 10-85%
acetonitrile/water (0.1% TFA)) to afford the title compound
N-{[2-[(4'-amino-2-biphenylyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2-
,3-dihydro-4-pyridazinyl]carbonyl}glycine (29 mg, 0.063 mmol, 45.3%
yield) as a white powder. 1H NMR (400 MHz, DMSO-d.sub.6) d ppm
15.86 (s, 1H), 10.11 (t, J=5.56 Hz, 1H), 7.19-7.36 (m, 5 H), 7.05
(d, J=7.07 Hz, 1H), 6.93 (d, J=7.33 Hz, 2H), 5.25 (s, 2H), 4.08 (d,
J=5.56 Hz, 2H), 3.16 (sept, J=6.82 Hz, 1H), 1.15 (d, J=6.82 Hz,
6H). MS (ES+) m/e 437 [M+H]+.
Example 98
##STR00109##
N-{[2-Cyclohexyl-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyridazin-
yl]carbonyl}glycine
98.sub.a) Ethyl-2-(cyclohexylhydrazono)-3-methylbutanoate. Ethyl
3-methyl-2-oxobutanoate (1 ml, 6.78 mmol) and DBU (1.23 ml, 8.16
mmol) were added to a solution of cyclohexylhydrazine hydrochloride
(1.23 g, 8.16 mmol) in Ethanol (10 ml) at room temperature. The
reaction was heated in the microwave at 150.degree. C. for 45 min.
The reaction was cooled and diluted with H.sub.2O and EtOAc. The
layers were separated and the aqueous layer backextracted with
EtOAc several times. The combined organic layers were washed with
1N HCl (2.times.) and Brine. The solution was dried (MgSO.sub.4),
filtered, and concentrated. The product was purified by column
chromatography (SiO.sub.2, 5-20% EtOAc/Hexanes) to give the title
compound as a colorless oil (1.21 g, 74%). 1H NMR (400 MHz,
DMSO-d.sub.6) d ppm 9.94 (d, J=5.05 Hz, 1H) 4.15 (q, J=7.24 Hz, 2H)
3.16-3.31 (m, J=13.89, 9.35, 4.80 Hz, 1H) 2.82 (sept, J=6.78 Hz,
1H) 1.78-1.92 (m, 2H) 1.46-1.73 (m, 3H) 1.26-1.38 (m, 4H) 1.24 (t,
J=7.07 Hz, 3H) 1.08-1.21 (m, 1H) 1.02 (d, J=6.82 Hz, 6H).
98b)
Ethyl-2-{cyclohexyl[3-(ethyloxy)-3-oxopropanoyl]hydrazono}-3-methylb-
utanoate. DBU (0.35 ml, 2.322 mmol) was added to a solution of the
compound from example 98a) (0.51 g, 2.122 mmol) in Tetrahydrofuran
(THF) (1 ml) at 0.degree. C. The reaction was brought to room
temperature and stirred for 10 minutes. The temperature was then
reduced to 0.degree. C. and ethyl malonyl chloride (0.32 ml, 2.374
mmol) was added. The reaction was brought to room temperature and
stirred for 2.5 h. 1N HCl was added and the solution diluted with
H.sub.2O and EtOAc. The layers were separated and aqueous phase
backextracted with EtOAc several times. The combined organic layers
were washed with Brine, dried (MgSO.sub.4), filtered and
concentrated. The product was purified by column chromatography
(SiO.sub.2, 10-25% EtOAc/Hexanes) to give a mixture of the title
compound and the cyclized product 55c) (126 mg, 13%). LCMS
(ES.sup.+) m/z 355.2, 309.2 (MH+).
98c) Ethyl
2-cyclohexyl-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyridazineca-
rboxylate. DBU (0.06 mL, 0.398 mmol) was added to a solution of the
compound from example 98b) (92 mg, 0.260 mmol) in 1,4-dioxane (1.5
ml) at room temperature. The reaction was heated in the microwave
at 130.degree. C. for 20 minutes. The reaction was cooled and
H.sub.2O and EtOAc were added. The layers were separated and the
aqueous phase backextracted with EtOAc several times. The combined
organic layers were washed with Brine, dried (MgSO.sub.4), filtered
and concentrated. The product was purified by column chromatography
(SiO.sub.2, 10-20% EtOAc/Hexanes) to give the title compound as a
colorless oil (36 mg, 45%). 1H NMR (400 MHz, DMSO-d.sub.6) d ppm
12.06 (s, 1H) 4.66 (tt, J=11.37, 4.04 Hz, 1H) 4.26 (q, J=7.07 Hz,
2H) 3.15 (sept, J=6.78 Hz, 1H) 1.51-1.89 (m, 7H) 1.29-1.44 (m, 2H)
1.26 (t, J=7.07 Hz, 3H) 1.16 (d, J=6.82 Hz, 6H) 1.08-1.22 (m,
1H).
98d) N-{[2-Cyclohexyl-5-hydroxy-6-(1
-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine.
Glycine, sodium salt (19 mg, 0.196 mmol) was added to a solution of
the compound from example 98c) (30 mg, 0.097 mmol) in
2-methoxyethanol (1 ml) at room temperature. The reaction was
heated to reflux and stirred for 2 h. The reaction was cooled and
diluted with H.sub.2O. The solution was filtered and 1N HCl added
to precipitate the product. The product was filtered and washed
with H.sub.2O and Hexanes to give the title compound as a white
solid (24 mg, 73%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm
15.77 (s, 1H) 13.01 (s, 1H) 10.35 (t, J=4.80 Hz, 1H) 4.67-4.83 (m,
1H) 4.11 (d, J=5.56 Hz, 2H) 3.18 (sept, J=6.86 Hz, 1H) 1.55-1.91
(m, 7H) 1.31-1.48 (m, 2H) 1.20 (d, J=6.82 Hz, 6H) 1.14-1.25 (m, 1
H).
Example 99
##STR00110##
N-{[2-Cyclohexyl-6-(1,1-dimethylethyl)-5-hydroxy-3-oxo-2,3-dihydro-4-pyrid-
azinyl]carbonyl}glycine
99a) Ethyl-2-(cyclohexylhydrazono)-3,3-dimethylbutanoate. To a
solution of trimethylpyruvic acid (0.6 g, 2.77 mmol) in MTBE (4.0
ml) was added DBU (0.58 ml, 3.85 mmol) followed by bromoethane (0.5
ml, 6.70 mmol) at room temperature. The reaction was heated in the
microwave at 120.degree. C. for 20 minutes. The reaction was cooled
and 10% NaHCO.sub.3 (aq) added. The layers were separated and the
organic layer was washed again with 10% NaHCO.sub.3 (aq). The
aqueous layer backextracted with Et.sub.2O several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The resulting oil was redissolved in
Ethanol (5.0 mL) and cyclohexylhydrazine hydrochloride (0.500 g,
3.32 mmol) and DBU (0.500 ml, 3.32 mmol) were added. The reaction
was heated in the microwave at 150.degree. C. for 25 minutes. The
reaction was cooled to room temperature and 1N HCl added. The
solution was diluted with H.sub.2O and EtOAc and the layers
separated. The aqueous layer was backextracted with EtOAc several
times. The combined organic layers were washed with Brine, dried
(MgSO.sub.4), filtered and concentrated. The product was purified
by column chromatography (SiO.sub.2, 5-15% EtOAc/Hexanes) to give
the title compound as a colorless oil (0.403 g, 57%). 1H NMR (400
MHz, DMSO-d.sub.6) d ppm 9.32 (d, J=4.80 Hz, 1H) 4.20 (q, J=7.07
Hz, 2H) 3.08-3.26 (m, 1H) 1.79-1.94 (m, 2H) 1.48-1.74 (m, 3H) 1.26
(t, J=7.07 Hz, 3H) 1.22-1.37 (m, 5H) 1.15 (s, 9H).
99b)
Ethyl-2-{cyclohexyl[3-(ethyloxy)-3-oxopropanoyl]hydrazono}-3,3-dimet-
hylbutanoate. DBU (0.21 ml, 1.393 mmol) was added to a solution of
the compound from example 99a) (0.323 g, 1.270 mmol) in
Tetrahydrofuran (THF) (1 ml) at 0.degree. C. The reaction was
brought to room temperature and stirred for 10 minutes. The
temperature was then reduced to 0.degree. C. and ethyl malonyl
chloride (0.19 ml, 1.410 mmol) was added. The reaction stirred for
2.5 h at room temperature followed by the addition of 1N HCl. The
solution was diluted with H.sub.2O and EtOAc and the layers
separated. The aqueous phase was backextracted with EtOAc several
times. The combined organic layers were washed with Brine, dried
(MgSO.sub.4), filtered and concentrated. The product was purified
by column chromatography (SiO.sub.2, 5-25% EtOAc/Hexanes) to give a
mixture of the title compound and the cyclized product 56c) (197
mg, 42%). LCMS (ES+) m/z 369.0, 323.0 (MH.sup.+).
99c) Ethyl
2-cyclohexyl-6-(1,1-dimethylethyl)-5-hydroxy-3-oxo-2,3-dihydro-4-pyridazi-
necarboxylate. DBU (0.11 ml, 0.730 mmol) was added to a solution of
the compound from example 99b) (173 mg, 0.470 mmol) in 1,4-dioxane
(2.5 ml) at room temperature. The reaction was heated to reflux and
stirred for 2 h. The reaction was cooled back to room temperature
and H.sub.2O was added. The solution was filtered and 1N HCl was
added to precipitate the product as a tan gum. The gum was filtered
and washed with H.sub.2O and Hexanes. The product was purified by
recrystallization from hot CH.sub.2Cl.sub.2 to give the title
compound as a white solid (103 mg, 77%). 1H NMR (400 MHz,
DMSO-d.sub.6) d ppm 12.39 (s, 1H) 4.65 (tt, J=11.49, 3.79 Hz, 1H)
4.27 (q, J=7.07 Hz, 2H) 1.52-1.87 (m, 7H) 1.33-1.44 (m, 2H) 1.32
(s, 9 H) 1.26 (t, J=7.07 Hz, 3H) 1.06-1.21 (m, 1H).
99d)
N-{[2-Cyclohexyl-6-(1,1-dimethylethyl)-5-hydroxy-3-oxo-2,3-dihydro-4-
-pyridazinyl]carbonyl}glycine. Glycine, sodium salt (58 mg, 0.598
mmol) was added to a solution of the compound from example 99c) (97
mg, 0.301 mmol) in 2-methoxyethanol (1.5 mL) at room temperature.
The reaction was heated to reflux and stirred for 1.5 h. The
reaction was then cooled to room temperature and H.sub.2O was
added. The solution was filtered and 1N HCl added to precipitate
the product. The product was filtered and redissolved in
CH.sub.2Cl.sub.2. The solution was concentrated under reduced
pressure and Hexanes added. The product was filtered to give the
title compound as an off-white solid (79 mg, 75%). 1H NMR (400 MHz,
DMSO-d.sub.6) .delta. ppm 16.27 (s, 1 H) 13.01 (s, 1H) 10.45 (t,
J=5.43 Hz, 1H) 4.66-4.83 (m, 1H) 4.12 (d, J=5.56 Hz, 2H) 1.57-1.89
(m, 7H) 1.36-1.52 (m, 2H) 1.34 (s, 9H) 1.05-1.30 (m, 1H).
Example 100
##STR00111##
N-{[5-Hydroxy-6-(1-methylethyl)-2-({3-[6-(4-methyl-1-piperazinyl)-3-pyridi-
nyl]phenyl}methyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 mL microwave tube was added
N-{[2-[(3-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 79, 31 mg, 0.073 mmol),
1-methyl-4-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridinyl]p-
iperazine (22.16 mg, 0.073 mmol), potassium carbonate (303 mg,
2.192 mmol), and tetrakis(triphenylphosphine)palladium (0) (2.53
mg, 2.192 .mu.mol) in 1,4-Dioxane (1.5 ml) and Water (0.500 ml).
The mixture was irradiated at 100.degree. C. for 20 minutes. The
reaction mixture was diluted with water (4 ml) and acidified with
1N HCl (1 ml) then filtered to remove any residue followed by
purification by HPLC chromatography (ODS silica, gradient 10-75%
acetonitrile/water (0.1% TFA)) to afford the title compound
N-{[5-hydroxy-6-(1
-methylethyl)-2-({3-[6-(4-methyl-1-piperazinyl)-3-pyridinyl]phenyl}methyl-
)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine (32 mg, 0.048
mmol, 65.7% yield) as a white powder. 1H NMR (400 MHz,
DMSO-d.sub.6) d ppm 15.88 (s, 1H), 12.98 (br. s., 1H), 10.18 (t,
J=5.56 Hz, 1 H), 9.71 (br. s., 1H), 8.46 (d, J=2.27 Hz, 1H), 7.91
(dd, J=8.84, 2.53 Hz, 1H), 7.52-7.61 (m, 2H), 7.42 (t, J=8.08 Hz,
1H), 7.23 (d, J=7.58 Hz, 1H), 7.07 (d, J=9.09 Hz, 1H), 5.32 (s,
2H), 4.48 (d, J=12.63 Hz, 2H), 4.09 (d, J=5.56 Hz, 2H), 3.52 (d,
J=10.36 Hz, 2H), 3.00-3.26 (m, 5H), 2.86 (d, J=4.04 Hz, 3H), 1.21
(d, J=6.82 Hz, 6H). MS (ES+) m/e 521 [M+H]+.
Example 101
##STR00112##
N-{[5-Hydroxy-6-(1-methylethyl)-3-oxo-2-({3-[2-(1-piperazinyl)-4-pyridinyl-
]phenyl}methyl)-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 mL microwave tube was added
N-{[2-[(3-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 79, 31 mg, 0.073 mmol),
1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridinyl]piperazine
(21.13 mg, 0.073 mmol), potassium carbonate (30.3 mg, 0.219 mmol),
and tetrakis(triphenylphosphine)palladium (0) (2.53 mg, 2.192
.mu.mol) in 1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture
was irradiated at 100.degree. C. for 20 minutes. The reaction
mixture was diluted with water (3 ml) and acidified with 1N HCl.
The mixture was then filtered to remove residual salts and purified
by HPLC chromatography (ODS silica, gradient 10-75%
acetonitrile/water (0.1% TFA)). The title compound,
N-{[5-hydroxy-6-(1-methylethyl)-3-oxo-2-({3-[2-(1-piperazinyl)-4-pyridiny-
l]phenyl}methyl)-2,3-dihydro-4-pyridazinyl]carbonyl}glycine (22 mg,
0.034 mmol, 46.2% yield), was obtained as a white solid tfa salt.
1H NMR (400 MHz, DMSO-d.sub.6) d ppm 15.88 (s, 1 H), 12.90 (br. s.,
1H), 10.17 (t, J=5.81 Hz, 1H), 8.75 (br. s., 2H), 8.22 (d, J=5.56
Hz, 1H), 7.72 (s, 1H), 7.70 (d, J=8.59 Hz, 1H), 7.48 (t, J=7.71 Hz,
1H), 7.35 (d, J=7.58 Hz, 1H), 7.14 (s, 1H), 7.01 (dd, J=5.31, 1.01
Hz, 1H), 5.34 (s, 2H), 4.09 (d, J=5.56 Hz, 2H), 3.75-3.84 (m, 4H),
3.13-3.27 (m, 5H), 1.21 (d, J=6.82 Hz, 6H). MS (ES+) m/e 507
[M+H]+.
Example 102
##STR00113##
N-{[5-Hydroxy-6-(1-methylethyl)-3-oxo-2-({2-[2-(1-piperazinyl)-4-pyridinyl-
]phenyl}methyl)-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
To a 5 mL microwave tube was added
N-{[2-[(2-bromophenyl)methyl]-5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihyd-
ro-4-pyridazinyl]carbonyl}glycine (example 78(b), 75 mg, 0.177
mmol),
1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridinyl]piperazine
(51.1 mg, 0.177 mmol), potassium carbonate (73.3 mg, 0.530 mmol),
and tetrakis(triphenylphosphine)palladium (0) (6.13 mg, 5.30
.mu.mol) in 1,4-Dioxane (1.5 ml) and Water (0.500 ml). The mixture
was irradiated at 100.degree. C. for 20 minutes. The reaction
mixture was diluted with water (3 ml) and 1N HCl (1 mL). The
mixture was then filtered to remove residual salts and purified by
HPLC chromatography (ODS silica, gradient 10-75% acetonitrile/water
(0.1% TFA)). The title compound,
N-{[5-hydroxy-6-(1-methylethyl)-3-oxo-2-({2-[2-(1
-piperazinyl)-4-pyridinyl]phenyl}methyl)-2,3-dihydro-4-pyridazinyl]carbon-
yl}glycine (52 mg. 0.083 mmol, 47.0% yield), was obtained as a
white solid (tfa salt) after trituration with ether. 1H NMR (400
MHz, DMSO-d.sub.6) d ppm 15.83 (s, 1 H), 13.02 (br. s., 1H), 10.07
(t, J=5.43 Hz, 1H), 8.79 (br. s., 2H), 8.19 (d, J=5.05 Hz, 1H),
7.39 (dd, J=5.68, 3.41 Hz, 2H), 7.24-7.31 (m, 1H), 7.21 (dd,
J=5.43, 3.66 Hz, 1H), 6.93 (s, 1H), 6.78 (d, J=5.05 Hz, 1H), 5.30
(s, 2H), 4.08 (d, J=5.56 Hz, 2H), 3.69-3.85 (m, 4H), 3.20 (br. s.,
4H), 3.12 (m, 1H), 1.12 (d, J=6.82 Hz, 6H). MS (ES+) m/e 507
[M+H]+.
Example 103
##STR00114##
2,2'-{{1-[(2-Chlorophenyl)methyl]-4-hydroxy-6-oxo-1,6-dihydropyridazine-3,-
5-diyl}bis[oxomethanediyl)imino]}diacetic acid
103a) Diethyl
4-hydroxy-6-oxo-1,6-dihydro-3,5-pyridazinedicarboxylate. Diethyl
ketomalonate (5 ml, 32.8 mmol) and catalytic AcOH (0.45 ml, 7.86
mmol) were added to a solution of ethyl-3-hydrazino-3-oxopropionate
(5.75 g, 39.3 mmol) in Ethanol (50 ml) at room temperature. The
reaction was heated to reflux and stirred for 3.5 h. The reaction
was then cooled to room temperature and solvent removed under
reduced pressure. The residue was redissolved in EtOAc and washed
with 1N HCl (2.times.). The aqueous phase was backextracted with
EtOAc several times. The combined organic layers were washed with
Brine, dried (MgSO.sub.4), filtered and concentrated to give a
yellow oil. The oil was dissolved in Ethanol (60 ml) and DBU (7.4
ml, 49.1 mmol) was added at room temperature. The reaction was
heated to reflux and stirred for 2 h. The reaction was cooled to
and H.sub.2O was added followed by 6N HCl to precipitate the
product. The suspension was cooled to 0.degree. C. and allowed to
set for 15 min. The solid was filtered and washed several times
with H.sub.2O and Hexanes. The product was purified by
recrystallization from hot EtOH to give the title compound as a
brown solid (1.73 g, 21%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta.
ppm 13.53 (s, 1H) 4.34 (q, J=7.24 Hz, 2H) 4.27 (q, J=7.07 Hz, 2H)
1.31 (t, J=7.20 Hz, 3H) 1.26 (t, J=7.20 Hz, 3H).
103b) Diethyl
1-[(2-chlorophenyl)methyl]-4-hydroxy-6-oxo-1,6-dihydro-3,5-pyridazinedica-
rboxylate. Sodium hydride (273 mg, 6.83 mmol) was added to a
solution of the compound from example 103a) (700 mg, 2.73 mmol) in
N,N-Dimethylformamide (DMF) (10 ml) at 0.degree. C. The reaction
was brought to room temperature and stirred for 30 minutes. The
temperature was then reduced to 0.degree. C. and 2-chlorobenzyl
bromide (0.36 ml, 2.77 mmol) was added. The reaction was brought to
room temperature and stirred for 3.5 h followed by the addition of
1N HCl. H.sub.2O and EtOAc were added and the layers separated. The
aqueous phase was backextracted with EtOAc several times. The
combined organic layers were washed with Brine, dried (MgSO.sub.4),
filtered and concentrated. The product was purified by column
chromatography (SiO.sub.2, 0-5% MeOH/CH.sub.2Cl.sub.2) to give the
title compound as a light yellow solid (681 mg, 66%). 1H NMR (400
MHz, DMSO-d.sub.6) .delta. ppm 7.48-7.53 (m, 1H) 7.24-7.42 (m, 2H)
7.03-7.15 (m, 1H) 5.34 (s, 2H) 4.33 (q, J=7.07 Hz, 2H) 4.27 (q,
J=7.07 Hz, 2H) 1.28 (t, J=7.07 Hz, 3H) 1.26 (t, J=7.07 Hz, 3H).
103c)
2,2'-{{1-[(2-Chlorophenyl)methyl]-4-hydroxy-6-oxo-1,6-dihydropyrida-
zine-3,5-diyl}bis[oxomethanediyl)imino]}diacetic acid. Glycine,
sodium salt (175 mg, 1.799 mmol) was added to a solution of the
compound from example 103b) (571 mg, 1.500 mmol) in Ethanol (10 ml)
at room temperature. The reaction was heated in the microwave at
120.degree. C. for 25 minutes. The reaction was cooled and solvent
removed under reduced pressure. H.sub.2O was added followed by 1N
HCl to precipitate the product. The solid was filtered then
redissolved in DMSO. The product was purified by HPLC (10-85%
ACN/H.sub.2O) to give the title compound as a white solid (13 mg,
2%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 16.26 (s, 1H) 12.99
(s, 1H) 12.75 (s, 1H) 10.00 (t, J=5.43 Hz, 1H) 8.85 (s, 1H) 7.51
(dd, J=7.71, 1.39 Hz, 1H) 7.27-7.39 (m, 2H) 7.10-7.16 (m, 1H) 5.40
(s, 2H) 4.11 (d, J=5.56 Hz, 2H) 3.95 (d, J=5.81 Hz, 2H).
Example 104
##STR00115##
N-{[5-Hydroxy-6-(1-methylethyl)-3-oxo-2-(2,2,2-trifluoroethyl)-2,3-dihydro-
-4-pyridazinyl]carbonyl}glycine
104a) Ethyl-3-methyl-2-[(2,2,2-trifluoroethyl)hydrazono]butanoate.
To a microwave tube was added ethyl-3-methyl-2-oxobutyrate (2.212
g, 15.34 mmol) and 2,2,2-trifluoroethylhydrazine (2.5 g, 15.34
mmol) in Ethanol (10 ml) and Acetic Acid (0.5 ml). The reaction was
irridatiated at 150.degree. C. for 20 minutes. The reaction mixture
was partitioned between water (15 ml) and ethyl acetate (25 ml) the
organic layer separated. After another extraction the organic
layers were combined, dried over MgSO4, filtered then evaporated
down to give a clear oil. The crude oil was purified by flash
column chromatography (5%-25% EtOAc:Hexanes) to give the product as
a clear oil
(ethyl-3-methyl-2-[(2,2,2-trifluoroethyl)hydrazono]butanoate (2.55
g, 10.51 mmol, 68.5% yield). 1H NMR (400 MHz, CHLOROFORM-d) d ppm
10.07 (s, 1H), 4.26 (q, J=7.07 Hz, 2H), 3.92 (dq, J=8.88, 5.18 Hz,
2H), 2.91 (sept, J=6.78 Hz, 1H), 1.35 (t, J=7.20 Hz, 3H), 1.10 (d,
J=6.82 Hz, 6H). MS (ES+) m/e 241 [M+H]+.
104b)
Ethyl-5-hydroxy-6-(1-methylethyl)-3-oxo-2-(2,2,2-trifluoroethyl)-2,-
3-dihydro-4-pyridazinecarboxylate. To a microwave tube was added
ethyl-3-methyl-2-[(2,2,2-trifluoroethyl)hydrazono]butanoate (2.5 g,
10.41 mmol) and Ethyl Malonyl Chloride (1.539 ml, 11.45 mmol) in
1,4-Dioxane (15 ml). The reaction was irridatiated at 150.degree.
C. for 20 minutes. The crude reaction mixture was evaporated down
to give a yellow oil. The crude oil was resuspended in 1,4-Dioxane
(20 ml) and DBU (1.726 ml, 11.45 mmol) was added. The solution was
irridatiated at 150.degree. C. for 20 minutes, diluted with water
(5 ml) and acidified with 6N HCl. The crude oil was purified by
flash column chromatography (5%-25% EtOAc: Hexanes). The desired
product was inseparable but present by LCMS and the material
carried on to the next step. ethyl 5-hydroxy-6-(1
-methylethyl)-3-oxo-2-(2,2,2-trifluoroethyl)-2,3-dihydro-4-pyridazinecarb-
oxylate (50 mg, 0.162 mmol, 1.559% yield). MS (ES+) m/e 309
[M+H]+.
104c)
N-{[5-Hydroxy-6-(1-methylethyl)-3-oxo-2-(2,2,2-trifluoroethyl)-2,3--
dihydro-4-pyridazinyl]carbonyl}glycine. To a 20 mL microwave tube
was added ethyl
5-hydroxy-6-(1-methylethyl)-3-oxo-2-(2,2,2-trifluoroethyl)-2,3-dihydro-4--
pyridazinecarboxylate (35 mg, 0.114 mmol) and Glycine Sodium Salt
(27.5 mg, 0.284 mmol) in 2-methoxyethanol (2 ml) and the mixture
was irradiated at 150.degree. C. for 20 minutes. The reaction
mixture was diluted with water (10 ml) and acidified with 1N HCl to
give a off-white precipitate that was collected by filtration and
washed with water, hexanes and ether to give
N-{[5-hydroxy-6-(1-methylethyl)-3-oxo-2-(2,2,2-trifluoroethyl)-2,-
3-dihydro-4-pyridazinyl]carbonyl}glycine (21 mg, 0.062 mmol, 54.3%
yield). 1H NMR (400 MHz, DMSO-d.sub.6) d ppm 16.25 (br. s., 1H),
13.01 (br. s., 1H), 10.01 (t, J=5.68 Hz, 1H), 4.95 (q, J=9.01 Hz,
2H), 4.12 (d, J=5.56 Hz, 2H), 3.12-3.25 (m, 1 H), 1.19 (d, J=6.82
Hz, 6H). MS (ES+) m/e 338 [M+H]+.
Example 105
##STR00116##
N-({2-[(2-Chlorophenyl)methyl]-6-[(ethyloxy)carbonyl]-5-hydroxy-3-oxo-2,3--
dihydro-4-pyridazinyl}carbonyl)glycine
Glycine, sodium salt (175 mg, 1.799 mmol) was added to a solution
of the compound from example 103b) (571 mg, 1.500 mmol) in Ethanol
(10 ml) at room temperature. The reaction was heated in the
microwave at 120.degree. C. for 25 minutes. The reaction was cooled
and solvent removed under reduced pressure. H.sub.2O was added
followed by 1N HCl to precipitate the product. The solid was
filtered then redissolved in DMSO. The product was purified by HPLC
(10-85% ACN/H.sub.2O) to give the title compound as a white solid
(75 mg, 12%). 1H NMR (400 MHz, DMSO-d.sub.6) .delta. ppm 13.01 (s,
1H) 9.96 (t, J=5.05 Hz, 1H) 7.51 (dd, J=7.71, 1.39 Hz, 1H)
7.27-7.40 (m, 2H) 7.17 (dd, J=7.58, 1.52 Hz, 1H) 5.41 (s, 2H) 4.32
(q, J=7.07 Hz, 2H) 4.10 (d, J=5.81 Hz, 2H) 1.28 (t, J=7.20 Hz,
3H).
Example 106
##STR00117##
N-{[2-{[2-Fluoro-4-(trifluoromethyl)phenyl]methyl}-5-hydroxy-6-(1-methylet-
hyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine
106a) Ethyl
2-{[2-fluoro-4-(trifluoromethyl)phenyl]methyl}-5-hydroxy-6-(1-methylethyl-
)-3-oxo-2,3-dihydro-4-pyridazinecarboxylate. To a solution of ethyl
5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinecarboxylate
(example 14(a), 0.5 g, 2.210 mmol) in N,N-Dimethylformamide (DMF)
(50 ml) at 0.degree. C. was added sodium hydride (0.133 g, 3.32
mmol) in portions. The reaction mixture was stirred at room
temperature for 45 minutes and then cooled back to 0.degree. C. and
1-(bromomethyl)-2-fluoro-4-(trifluoromethyl)benzene (0.568 g, 2.210
mmol) was added portionwise. The mixture was stirred at ambient
temperature for 2.5 hours then quenched with 1N HCl (2 ml) and
diluted with water (10 ml). The aqueous solution was extracted with
ethyl acetate (2.times.30 ml), the organic layers combined and
washed with water (100 ml) and brine (100 ml), dried over Magnesium
sulfate, filtered and solvents removed with rotary evaporation. The
crude oil was purified by flash column chromatography (5-100% ethyl
acetate in hexanes) to provide the title compound (ethyl
2-{[2-fluoro-4-(trifluoromethyl)phenyl]methyl}-5-hydroxy-6-(1-methylethyl-
)-3-oxo-2,3-dihydro-4-pyridazinecarboxylate (335 mg, 0.749 mmol,
33.9% yield) as a clear oil. 1H NMR (400 MHz, DMSO-d.sub.6) dppm
12.36 (s, 1H), 7.67-7.73 (m, 1H), 7.58 (d, J=8.84 Hz, 1H), 7.45 (t,
J=7.58 Hz, 1H), 5.27 (s, 2H), 4.27 (q, J=7.07 Hz, 2H), 3.15 (sept,
J=6.78 Hz, 1H), 1.26 (t, J=7.07 Hz, 3H), 1.13 (d, J=6.82 Hz, 6H).
MS (ES+) m/e 403 [M+H]+.
106b)
N-{[2-{[2-Fluoro-4-(trifluoromethyl)phenyl]methyl}-5-hydroxy-6-(1
-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine. To
a 5 mL microwave tube was added ethyl
2-{[2-fluoro-4-(trifluoromethyl)phenyl]methyl}-5-hydroxy-6-(1
-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinecarboxylate (275 mg,
0.684 mmol) and Glycine Sodium Salt (166 mg, 1.709 mmol) in
2-methoxyethanol (2 ml) and the mixture was irradiated at
150.degree. C. for 20 minutes. The crude reaction mixture turned a
violet blue. The mixture was diluted with water (10 ml) and
acidified with 1N HCl. The aqueous solution was then extracted with
ethyl acetate (40 mL), organics dried over MgSO4, filtered and
solvents removed by evaporation. The crude blue oil was purified by
HPLC chromatography (ODS silica, gradient 25-95% acetonitrile/water
(0.1% TFA)) to afford the title compound,
N-{[2-{[2-fluoro-4-(trifluoromethyl)phenyl]methyl}-5-hydroxy-6-(1-methyle-
thyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}glycine (45 mg,
0.103 mmol, 15.11% yield), as a white solid. 1H NMR (400 MHz,
DMSO-d.sub.6) d ppm 15.94 (s, 1H), 12.97 (br. s., 1H), 10.09 (t,
J=5.18 Hz, 1H), 7.72 (d, J=10.11 Hz, 1H), 7.54-7.62 (m, 1H), 7.49
(t, J=7.45 Hz, 1H), 5.38 (s, 2H), 4.09 (d, J=5.56 Hz, 2H), 3.17
(sept, J=6.82 Hz, 1H), 1.17 (d, J=6.82 Hz, 6 H). MS (ES+) m/e 432
[M+H]+.
Example 107
##STR00118##
N-{[5-Hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}g-
lycine
To a 100 mL round bottom flask was added ethyl
5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinecarboxylate
(example 14(a), 55 mg, 0.243 mmol) and Glycine Sodium Salt (59.0
mg, 0.608 mmol) in 2-methoxyethanol (10 ml) and the mixture was
refluxed at 130.degree. C. for 2 hours. The mixture was diluted
with water (10 ml) and acidified with 1N HCl. The aqueous solution
was then extracted with ethyl acetate (40 mL), organics dried over
MgSO4, filtered and solvents removed by evaporation. The crude oil
was purified by HPLC chromatography (ODS silica, gradient 25-95%
acetonitrile/water (0.1% TFA)) to afford the title compound,
N-{[5-hydroxy-6-(1-methylethyl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}-
glycine (45 mg, 0.175 mmol, 71.8% yield), as a white solid. 1H NMR
(400 MHz, DMSO-d.sub.6) d ppm 15.74 (br. s., 1H), 13.07 (s, 1H),
10.19 (t, J=5.68 Hz, 1H), 4.11 (d, J=5.81 Hz, 2H), 3.14 (sept,
J=6.82 Hz, 1H), 1.17 (d, J=6.82 Hz, 6H). MS (ES+) m/e 256
[M+H]+.
Example 108
##STR00119##
N-{[5-Hydroxy-6-(1H-indol-3-yl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}g-
lycine
To a microwave tube was added ethyl 1H-indol-3-yl(oxo) acetate (1
g, 4.60 mmol) and Ethyl Malonyl hydrazine (0.807 g, 5.52 mmol) in
Ethanol (5 ml) and Acetic Acid (0.5 ml). The reaction was
irridatiated at 150.degree. C. for 20 minutes. The crude reaction
mixture was evaporated down to give a yellow oil. The crude oil was
resuspended in 1,4-Dioxane (20 ml) and DBU (1.041 ml, 6.91 mmol)
was added. The solution was irridatiated at 150.degree. C. for 20
minutes, diluted with water (5 ml) and acidified with 6N HCl to
cause a precipitate. The orange precipitate was collected by
filtration and dried. The precipitate was determined to contain the
desired intermediate ester by LCMS and carried on into the next
reaction without purification. To a 5 mL microwave tube, a mixture
of the crude intermediate ester (50 mg, 0.167 mmol), Glycine Sodium
Salt (40.5 mg, 0.418 mmol) and 2-methoxyethanol (2 ml) was refluxed
at 150.degree. C. for 20 minutes. The mixture was diluted with
water (10 ml) and acidified with 1N HCl. The aqueous solution was
then extracted with ethyl acetate (40 mL), organics dried over
MgSO4, filtered and solvents removed by evaporation. The crude oil
was purified by HPLC chromatography (ODS silica, gradient 10-85%
acetonitrile/water (0.1% TFA)) to afford the title compound,
N-{[5-hydroxy-6-(1H-indol-3-yl)-3-oxo-2,3-dihydro-4-pyridazinyl]carbonyl}-
glycine (11 mg, 0.032 mmol, 19.05% yield), as a yellow solid. 1H
NMR (400 MHz, DMSO-d.sub.6) d ppm 13.23 (s, 1H), 12.99 (br. s.,
1H), 11.47-11.77 (m, 1H), 10.22-10.55 (m, 1H), 8.02-8.37 (m, 2H),
7.48 (d, J=8.08 Hz, 1H), 7.17-7.24 (m, 1H), 7.09-7.17 (m, 1H), 4.16
(d, J=5.56 Hz, 2H). MS (ES+) m/e 329 [M+H]+.
Example 109
##STR00120##
N-{[5-Hydroxy-3-oxo-6-(2-thienyl)-2,3-dihydro-4-pyridazinyl]carbonyl}glyci-
ne
109a) Ethyl 3-{(2E)-2-[2-(ethyloxy)-2-oxo-1-(2-thienyl)
ethylidene]hydrazino}-3-oxopropanoate. To a microwave tube was
added ethyl oxo(2-thienyl)acetate (2 g, 10.86 mmol) and Ethyl
Malonyl hydrazine (1.904 g, 13.03 mmol) in Ethanol (5 ml) and
Acetic Acid (0.5 ml). The reaction was irridatiated at 150.degree.
C. for 20 minutes. The crude reaction mixture was evaporated down
to give a yellow oil. The crude oil was purified by flash column
chromatography (5%-25% EtOAc: hexanes) to give the product as a
clear oil, ethyl
3-{(2E)-2-[2-(ethyloxy)-2-oxo-1-(2-thienyl)ethylidene]hydrazino}-3-oxopro-
panoate (0.6 g, 1.921 mmol, 17.69% yield). 1H NMR (400 MHz,
DMSO-d.sub.6) d ppm 11.52 (s, 1H), 7.67 (dd, J=5.05, 1.01 Hz, 1H),
7.39 (dd, J=3.79, 1.01 Hz, 1H), 7.11 (dd, 1H), 4.41 (q, J=7.24 Hz,
2H), 4.11 (q, J=7.24 Hz, 2 H), 3.69 (s, 2H), 1.33 (t, J=7.07 Hz,
3H), 1.19 (t, J=7.07 Hz, 3H). MS (ES+) m/e 313 [M+H]+.
109b) Ethyl
5-hydroxy-3-oxo-6-(2-thienyl)-2,3-dihydro-4-pyridazinecarboxylate.
To a microwave tube was added ethyl
3-{(2E)-2-[2-(ethyloxy)-2-oxo-1-(2-thienyl)ethylidene]hydrazino}-3-oxopro-
panoate (600 mg, 1.921 mmol) and potassium carbonate (133 mg, 0.960
mmol) in Ethanol (2 ml). The reaction was irridatiated at
150.degree. C. for 20 minutes. The crude reaction mixture was
diluted with water (10 ml) and acidified with 1N HCl which gave a
precipitate. The precipitate was filtered and dried to give the
product, ethyl
5-hydroxy-3-oxo-6-(2-thienyl)-2,3-dihydro-4-pyridazinecarboxylate
(450 mg, 1.690 mmol, 88% yield) as an off white powder. 1H NMR (400
MHz, DMSO-d.sub.6) d ppm 13.02 (s, 1H), 7.83 (dd, J=3.66, 1.14 Hz,
1H), 7.65 (dd, J=5.05, 1.26 Hz, 1H), 7.15 (dd, J=5.05, 3.79 Hz,
1H), 4.31 (q, J=7.07 Hz, 2H), 1.29 (t, J=7.07 Hz, 3H). MS (ES+) m/e
267 [M+H]+.
109c)
N-{[5-Hydroxy-3-oxo-6-(2-thienyl)-2,3-dihydro-4-pyridazinyl]carbony-
l}glycine. To a 5 mL microwave tube was added ethyl
5-hydroxy-3-oxo-6-(2-thienyl)-2,3-dihydro-4-pyridazinecarboxylate
(60 mg, 0.225 mmol) and Glycine Sodium Salt (54.7 mg, 0.563 mmol)
in 2-methoxyethanol (2 ml) and the mixture was irradiated at
150.degree. C. for 20 minutes. The mixture was diluted with water
(10 ml) and acidified with 1N HCl. The precipitate was collected by
filtration and recrystallized in ethanol to afford the title
compound,
N-{[5-hydroxy-3-oxo-6-(2-thienyl)-2,3-dihydro-4-pyridazinyl]carbonyl}glyc-
ine (30 mg, 0.101 mmol, 44.6% yield), as an off white solid. 1H NMR
(400 MHz, DMSO-d.sub.6) d ppm 13.41 (s, 1H), 13.00 (s, 1H), 10.25
(t, J=5.81 Hz, 1H), 7.79-7.97 (m, 1H), 7.70 (d, J=4.55 Hz, 1H),
7.18 (dd, J=5.05, 3.79 Hz, 1H), 4.16 (d, J=5.81 Hz, 2H). MS (ES+)
m/e 296 [M+H]+.
BIOLOGICAL BACKGROUND
The following references set out information about the target
enzymes, HIF prolyl hydroxylases, and methods and materials for
measuring inhibition of same by small molecules. M. Hirsila, P.
Koivunen, V. Gunzler, K. I. Kivirikko, and J. Myllyharju
"Characterization of the Human Prolyl 4-Hydroxylases That Modify
the Hypoxia-inducible Factor" J. Biol. Chem., 2003, 278,
30772-30780. C. Willam, L. G. Nicholls, P. J. Ratcliffe, C. W.
Pugh, P. H. Maxwell "The prolyl hydroxylase enzymes that act as
oxygen sensors regulating destruction of hypoxia-inducible factor
.alpha." Advan. Enzyme Regul., 2004, 44, 75-92 M. S. Wiesener, J.
S. Jurgensen, C. Rosenberger, C. K. Scholze, J. H. Horstrup, C.
Warnecke, S. Mandriota, I. Bechmann, U. A. Frei, C. W. Pugh, P. J.
Ratcliffe, S. Bachmann, P. H. Maxwell, and K.-U. Eckardt
"Widespread hypoxia-inducible expression of HIF-2.alpha. in
distinct cell populations of different organs" FASEB J, 2003, 17,
271-273. S. J. Klaus, C. J. Molineaux, T. B. Neff, V.
Guenzler-Pukall, I. Lansetmo Parobok, T. W. Seeley, R. C.
Stephenson "Use of hypoxia-inducible factor .alpha. (HIF.alpha.)
stabilizers for enhancing erythropoiesis" PCT Int. Appl. (2004), WO
2004108121 A1 C. Warnecke, Z. Zaborowska, J. Kurreck, V. A.
Erdmann, U. Frei, M. Wiesener, and K.-U. Eckardt "Differentiating
the functional role of hypoxia-inducible factor (HIF)-1.alpha. and
HIF-2.alpha. (EPAS-1) by the use of RNA interference:
erythropoietin is a HIF-2.alpha. target gene in Hep3B and Kelly
cells" FASEB J., 2004, 18, 1462-1464. For the expression of EGLN3
see: R. K. Bruick and S. L. McKnight "A Conserved Family of
Prolyl-4-Hydroxylases That Modify HIF" Science, 2001, 294,
1337-1340. For the expression of HIF2.alpha.-CODD see: a) P.
Jaakkola, D. R. Mole, Y.-M. Tian, M. I. Wilson, J. Gielbert, S. J.
Gaskell, A. von Kriegsheim, H. F. Hebestreit, M. Mukherji, C. J.
Schofield, P. H. Maxwell, C. W. Pugh, P, J. Ratcliffe "Targeting of
HIF-.alpha. to the von Hippel-Lindau Ubiquitylation Complex by
O.sub.2-Regulated Prolyl Hydroxylation" Science, 2001, 292,
468-472. b) M. Ivan, K. Kondo, H. Yang, W. Kim, J. Valiando, M.
Ohh, A. Salic, J. M. Asara, W. S. Lane, W. G. Kaelin Jr.
"HIF.alpha. Targeted for VHL-Mediated Destruction by Proline
Hydroxylation: Implications for O.sub.2 Sensing" Science, 2001,
292, 464-468. For the expression of VHL, elongin b and elongin c
see: A. Pause, S. Lee, R. A. Worrell, D. Y. T. Chen, W. H. Burgess,
W. M. Linehan, R. D. Klausner "The von Hippel-Lindau
tumor-suppressor gene product forms a stable complex with human
CUL-2, a member of the Cde53 family of proteins" Proc. Natl. Acad.
Sci. USA, 1997, 94, 2156-2161. Biological Assay(s) EGLN3 Assay
Materials:
His-MBP-EGLN3 (6HisMBPAttB1EGLN3(1-239)) was expressed in E. Coli
and purified from an amylase affinity column. Biotin-VBC
[6HisSumoCysVHL(2-213), 6HisSumoElonginB(1-118), and
6HisSumoElonginC(1-112)] and His-GB1-HIF2.alpha.-CODD
(6HisGB1tevHIF2A (467-572)) were expressed from E. Coli.
Method:
Cy5-labelled HIF2.alpha. CODD, and a biotin-labeled VBC complex
were used to determine EGLN3 inhibition. EGLN3 hydroxylation of the
Cy5CODD substrate results in its recognition by the biotin-VBC.
Addition of a Europium/streptavidin (Eu/SA) chelate results in
proximity of Eu to Cy5 in the product, allowing for detection by
energy transfer. A ratio of Cy5 to Eu emission (LANCE Ratio) is the
ultimate readout, as this normalized parameter has significantly
less variance than the Cy5 emission alone.
Then 50 nL of inhibitors in DMSO (or DMSO controls) were stamped
into a 384-well low volume Corning NBS plate, followed by addition
of2.5 .mu.L of enzyme [50 mL buffer (50 mM HEPES/50 mM KCl)+1 mL of
a 10 mg/mL BSA in buffer+6.25 .mu.L of a 10 mg/mL FeCl.sub.2
solution in water+100 .mu.L of a 200 mM solution of ascorbic acid
in water+15.63 .mu.L EGLN3] or control [50 mL buffer+1 mL of a 10
mg/mL BSA in buffer+6.25 .mu.L of a 10 mg/mL FeCl.sub.2 solution in
water+100 .mu.L of a 200 mM solution of ascorbic acid in water].
Following a 3 minutes incubation, 2.5 .mu.L of substrate [50 mL
Buffer+68.6 .mu.L biotin-VBC+70.4 .mu.L Eu (at 710 .mu.g/mL
stock)+91.6 .mu.L Cy5CODD+50 .mu.L of a 20 mM solution of
2-oxoglutaric acid in water+0.3 mM CHAPS] was added and incubated
for 30 minutes. The plate was loaded into a PerkinElmer Viewlux for
imaging. For dose response experiments, normalized data were fit by
ABASE/XC50 using the equation y=a+(b-a)/(1+(10^x/10^c)^d), where a
is the minimum % activity, b is the maximum % activity, c is the
pIC.sub.50, and d is the Hill slope.
The IC.sub.50 for exemplified compounds in the EGLN3 assay ranged
from approximately 1-3200 nanomolar. This range represents the data
accumulated as of the time of the filing of this initial
application. Later testing may show variations in IC.sub.50 data
due to variations in reagents, conditions and variations in the
method(s) used from those given herein above. So this range is to
be viewed as illustrative, and not a absolute set of numbers.
Measure Epo Protein Produced by Hep3B Cell Line Using ELISA
Method.
Hep3B cells obtained from the American Type Culture Collection
(ATCC) are seeded at 2.times.10^4 cells/well in Dulbecco's Modified
Eagle Medium (DMEM)+10% FBS in 96-well plates. Cells are incubated
at 37 deg C./5% CO2/90% humidity (standard cell culture incubation
conditions). After overnight adherence, medium is removed and
replaced with DMEM without serum containing test compound or DMSO
negative control. Following 48 hours incubation, cell culture
medium is collected and assayed by ELISA to quantitate Epo
protein.
Measure Epo Protein Produced by Hep3B Cell Line Using AlphaLISA
Method.
Hep3B cells obtained from the American Type Culture Collection
(ATCC) are seeded at 2.times.10^4 cells/well in Dulbecco's Modified
Eagle Medium (DMEM)+10% FBS in 96-well plates. Cells are incubated
at 37 deg C./5% CO2/90% humidity (standard cell culture incubation
conditions). After overnight adherence, medium is removed and
replaced with DMEM with 2% serum containing test compound or DMSO
negative control. Following 48 hours incubation, cell culture
medium is collected and either frozen for later assay or assayed
immediately by AlphaLISA to quantitate Epo protein.
The EC.sub.50 for exemplar compounds in the Hep3B ELISA and
AlphaLISA assay ranged from approximately 0.4-100 micromolar,
except example 26, using the reagents and under the conditions
outlined herein above. Example 26 has demonstrated an EC.sub.50 in
the Hep3B ELISA assay of greater than 100 micromolar, the maximum
concentration tested. This range represents the data accumulated as
of the time of the filing of this initial application. Later
testing may show variations in EC.sub.50 data due to variations in
reagents, conditions and variations in the method(s) used from
those given herein above. So this range is to be viewed as
illustrative, and not a absolute set of numbers.
These compound are believed to be useful in therapy as defined
above and to not have unacceptable or untoward effects when used in
compliance with a permitted therapeutic regime.
The foregoing examples and assay have been set forth to illustrate
the invention, not limit it. What is reserved to the inventors is
to be determined by reference to the claims.
* * * * *