U.S. patent application number 13/742633 was filed with the patent office on 2013-05-16 for bicyclo-substituted pyrazolon azo derivatives, preparation process and pharmaceutical use thereof.
This patent application is currently assigned to SHANGHAI HENGRUI PHARMACEUTICAL CO. LTD.. The applicant listed for this patent is JIANGSU HENGRUI MEDICINE CO. LTD., SHANGHAI HENGRUI PHARMACEUTICAL CO. LTD.. Invention is credited to Yiqian Chen, Hongbo Fei, Hejun Lu, Peng Cho Tang, Li Wang, Shenglan Wang, Hao Zheng.
Application Number | 20130123507 13/742633 |
Document ID | / |
Family ID | 40878665 |
Filed Date | 2013-05-16 |
United States Patent
Application |
20130123507 |
Kind Code |
A1 |
Tang; Peng Cho ; et
al. |
May 16, 2013 |
BICYCLO-SUBSTITUTED PYRAZOLON AZO DERIVATIVES, PREPARATION PROCESS
AND PHARMACEUTICAL USE THEREOF
Abstract
The bicyclo-substituted pyrazolon-azo derivatives of formula (I)
or pharmaceutical acceptable salts, hydrates or solvates thereof,
methods for their preparation, pharmaceutical compositions
containing the same and their use as a therapeutic agent,
especially as thrombopoietin (TPO) mimetics and their use as
agonists of thrombopoietin receptor are disclosed. The definition
of substituents in formula (I) are the same as defined in the
description. ##STR00001##
Inventors: |
Tang; Peng Cho; (Shanghai,
CN) ; Lu; Hejun; (Shanghai, CN) ; Zheng;
Hao; (Shanghai, CN) ; Chen; Yiqian; (Shanghai,
CN) ; Fei; Hongbo; (Shanghai, CN) ; Wang;
Shenglan; (Shanghai, CN) ; Wang; Li;
(Shanghai, CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
JIANGSU HENGRUI MEDICINE CO. LTD.;
LTD.; SHANGHAI HENGRUI PHARMACEUTICAL CO. |
Lianyungang
Shanghai |
|
CN
CN |
|
|
Assignee: |
SHANGHAI HENGRUI PHARMACEUTICAL CO.
LTD.
Shanghai
CN
JIANGSU HENGRUI MEDICINE CO. LTD.
Lianyungang
CN
|
Family ID: |
40878665 |
Appl. No.: |
13/742633 |
Filed: |
January 16, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
12812119 |
Jul 8, 2010 |
8367710 |
|
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13742633 |
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Current U.S.
Class: |
548/201 ;
548/312.4; 548/364.1; 548/364.4; 548/365.7; 548/367.4 |
Current CPC
Class: |
A61P 7/00 20180101; A61P
43/00 20180101; C07D 403/12 20130101; C07D 231/22 20130101; A61P
7/04 20180101; A61P 7/06 20180101; C07D 417/12 20130101; C07D
405/12 20130101; C07D 409/12 20130101; C07D 405/10 20130101; C07D
231/38 20130101; C07D 231/08 20130101; C07D 405/04 20130101 |
Class at
Publication: |
548/201 ;
548/367.4; 548/364.1; 548/364.4; 548/365.7; 548/312.4 |
International
Class: |
C07D 417/12 20060101
C07D417/12; C07D 409/12 20060101 C07D409/12; C07D 405/10 20060101
C07D405/10; C07D 231/08 20060101 C07D231/08; C07D 403/12 20060101
C07D403/12 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 10, 2008 |
CN |
CN 200810000346.6 |
Jan 4, 2009 |
CN |
PCT/CN2009/000001 |
Claims
1-13. (canceled)
14. A compound of formula (IA) ##STR00148## wherein: A is selected
from the group consisting of carbon and oxygen; R.sup.5, R.sup.6,
and R.sup.7 are each independently selected from the group
consisting of hydrogen, alkyl, alkoxy, halogen, hydroxyl, amino,
nitro, cyano, carboxylic acid, and carboxylic ester; R.sup.8,
R.sup.9, R.sup.10, and R.sup.11 are each independently selected
from the group consisting of hydrogen and alkyl, wherein R.sup.8,
R.sup.9, R.sup.10, and R.sup.11 can substitute any open valence of
the ring to which they attach; and n is 0, 1, or 2.
15. The compound according to claim 14, wherein R.sup.5, R.sup.6,
and R.sup.7 are hydrogen.
16. The compound according to claim 14, wherein R.sup.8, R.sup.9,
R.sup.10, and R.sup.11 are each independently hydrogen, methyl, and
ethyl.
17. The compound according to claim 14, which is selected from the
group consisting of: ##STR00149##
18. A process for preparing the compound of claim 14, comprising:
##STR00150## converting an amino substituted benzocycle of formula
(IA_a), in the presence of a carbonyl compound, to the
corresponding compound of formula (IA).
19. The process according to claim 18, wherein the converting step
comprises converting the amino substituted benzocycle of formula
(IA_a) to the corresponding diazo substituted benzocycle;
##STR00151## reducing the diazo substituted benzocycle to obtain a
hydrazine of formula (IA_b); and coupling the hydrazine of formula
(IA_b) with the carbonyl compound to obtain the compound of formula
(IA).
Description
FIELD
[0001] This disclosure relates to novel bicyclo-substituted
pyrazolon-azo derivatives represented by formula (I), methods for
their preparation, pharmaceutical compositions containing the same,
and their use as a therapeutic agent, particularly as
thrombopoietin (TPO) mimetics and their use as agonists of the
thrombopoietin receptor.
BACKGROUND
[0002] Thrombopoietin (TPO), also called megakaryocyte growth and
development factor (MGDF), thrombocytopoiesis stimulating factor
(TSF), c-myeloproliferative leukemia ligand (c-Mpl), mpl ligand, or
megapoietin, is a glycoprotein that has been shown to be involved
in the production of platelets. See Wendling, F., et. al.,
Biotherapy 10(4): 269-77 (1998); Kuter D. I. et al., The
Oncologist, 1: 98-106 (1996); Metcalf, Nature 369: 519-520
(1994).
[0003] Under certain circumstances, the activity of TPO results
from the binding of TPO with the TPO receptor (also called MPL).
The TPO receptor has been cloned and its amino acid sequence has
been described. See Vigon et al., Proc. Nat. Acad. Sci., 89:
5640-5644 (1992).
[0004] TPO is a 332-amino acid glycosylated polypeptide that plays
a key role in the regulation of megakaryocytopoiesis, and in the
process in which platelets are produced by bone marrow
megakaryocytes. See Kuter et al., Proc. Natl. Acad. Sci. USA 91:
11104-11108 (1994); Barley et al., Cell 77:1117-1124 (1994);
Kaushansky et al., Nature 369:568-571 (1994); Wendling et al.,
Nature 369: 571-574 (1994); and Sauvage et al., Nature 369: 533-538
(1994). TPO is produced in the liver but functions mainly in the
bone marrow, where it stimulates the differentiation of stem cells
into megakaryocyte progenitors, and stimulates megakaryocyte
proliferation, polyploidization and, ultimately, enters the
platelet circulation in the body. TPO is also a primary regulator
in situations involving thrombocytopenia and in a number of studies
that include increasing platelet counts, platelet size and isotope
incorporation into platelets of recipient animals. See, Metcalf
Nature 369: 519-520 (1994). Specifically, TPO is considered to
affect megakaryocytopoiesis by several ways: (1) it causes increase
in megakaryocyte size and number; (2) it increases DNA contents,
the forms of polyploidy, and the number of megakaryocytes; (3) it
increases megakaryocyte endomitosis; (4) it increases the number of
mature megakaryocytes; (5) it increases the percentage of precursor
cells, the number of small acetylcholinesterase positive cells, the
number of bone marrow cells.
[0005] Platelets are necessary for blood clotting. When platelet
numbers are very low, a patient is at risk of death from
catastrophic hemorrhage. Thus, TPO has been used for both the
diagnosis and the treatment of various hematological disorders, for
example, diseases primarily caused by platelet defects. Likewise,
TPO may be useful for the treatment of thrombocytopenic conditions,
especially those derived from chemotherapy, radiation therapy, or
bone marrow transplantation for the treatment of cancer or
lymphoma.
[0006] Because the slow recovery of platelet levels in patients
suffering from thrombocytopenia is a serious problem, it would be
desirable to provide a compound for the treatment of
thrombocytopenia by acting as a TPO mimetic. A few years ago, the
development of TPO peptide mimetics was reported (WO96/40750,
WO98/25965). These peptides were designed to bind and activate the
TPO receptor (TPO-R) but have no sequence homology to the natural
TPO. In recent years, a number of active small-molecule TPO
mimetics have been reported, including 1,4-benzodiazepin-2-ones
(JP11001477), metal complexes derived from Schiff base ligands
(WO99/11262), cyclic polyamine derivatives (WO00/28987),
thiazol-2-yl-benzamides (WO01/07423, WO01/53267), azo-aryl
derivatives (WO00/35446, WO01/17349), 2-aryl-naphthimidazoles
(WO01/39773, WO01/53267), and semicarbazone derivatives
(WO01/34585). In cell-based systems, all of these molecules can
activate signal transduction pathways that are dependent on the
presence of the TPO receptor on the cell membrane. Certain types of
compounds can directly act on the TPO receptor itself. It has been
reported that certain substituted thiosemicarbazone derivatives are
actually effective agonists of the TPO receptor. Some of the most
preferred compounds of this series were found to stimulate the
proliferation and differentiation of TPO-responsive human cell
lines and TPO in human bone marrow cultures that has a
concentration below 100 nM.
[0007] Several patents assigned on their face to GlaxoSmithKline
describe a thrombopoietin analog, eltrombopag (WO00/189457,
WO01/089457, WO2006/064957), which shows good activity.
SUMMARY
[0008] The present disclosure describes compounds that are TPO
receptor agonists and TPO mimetics.
[0009] The present disclosure is directed to compounds of
bicyclo-substituted pyrazolon-azo derivatives of formula (I) and
tautomers, enantiomers, diastereomers, racemics, pharmaceutically
acceptable salts, hydrates or solvates, as well as their
metabolites, metabolic precursors or prodrugs thereof.
##STR00002##
[0010] Wherein:
[0011] A is selected from the group consisting of carbon and
oxygen;
[0012] R is selected from the group consisting of hydrogen and
alkyl;
[0013] R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are each independently
selected from the group consisting of hydrogen, alkyl, alkoxy,
halogen, aryl and heteroaryl, wherein the aryl or heteroaryl is
each optionally substituted with one or more groups selected from
the group consisting of alkyl, halogen, hydroxyl, tetrazolyl,
imidazolyl, dihydroimidazolyl, carboxylic acid and carboxylic
ester;
[0014] R.sup.5, R.sup.6 and R.sup.7 are each independently selected
from the group consisting of hydrogen, alkyl, alkoxy, halogen,
hydroxyl, amino, nitro, cyano, carboxylic acid and carboxylic
ester;
[0015] R.sup.8, R.sup.9, R.sup.10 and R.sup.11 are each
independently selected from the group consisting of hydrogen and
alkyl; and
[0016] n is 0, 1 or 2.
[0017] In some preferred embodiments of present disclosure:
[0018] A is selected from the group consisting of carbon and
oxygen;
[0019] R is selected from the group consisting of hydrogen and
alkyl;
[0020] R.sup.1 is selected from the group consisting of aryl and
heteroaryl, wherein the aryl or heteroaryl is each optionally
substituted with one or more groups selected from the group
consisting of alkyl, halogen, hydroxyl, tetrazolyl, imidazolyl,
dihydroimidazolyl, carboxylic acid and carboxylic ester;
[0021] R.sup.2, R.sup.3 and R.sup.4 are each independently selected
from the group consisting of hydrogen, alkyl, alkoxy and
halogen;
[0022] R.sup.5, R.sup.6 and R.sup.7 are each independently selected
from the group consisting of hydrogen, alkyl, alkoxy, halogen,
hydroxyl, amino, nitro, cyano, carboxylic acid and carboxylic
ester;
[0023] R.sup.8, R.sup.9, R.sup.10 and R.sup.11 are each
independently selected from the group consisting of hydrogen and
alkyl; and
[0024] n is 0, 1 or 2.
[0025] The compounds of formula (I) of the present disclosure
preferably include, but are not limited to:
TABLE-US-00001 Example No. Structure Name 1 ##STR00003##
2'-Hydroxy-3'-[N'-(1-indan-5-yl-3-
methyl-5-oxo-1,5-dihydro-pyrazol- 4-ylidene)-hydrazino]-biphenyl-3-
carboxylic acid 2 ##STR00004## 5'-Fluoro-2'-hydroxy-3'-[N'-(1-
indan-5-yl-3-methyl-5-oxo-1,5- dihydro-pyrazol-4-ylidene)-
hydrazino]-biphenyl-3- carboxylic acid 3 ##STR00005##
2'-Hydroxy-3'-{N'-[3-methyl-5- oxo-1-(5,6,7,8-tetrahydro-
naphthalen-2-yl)-1,5-dihydro- pyrazol-4-ylidene]-hydrazino}-
biphenyl-3-carboxylic acid 4 ##STR00006##
5'-Fluoro-2'-hydroxy-3'-{N'-[3- methyl-5-oxo-1-(5,6,7,8-
tetrahydro-naphthalen-2-yl)-1,5- dihydro-pyrazol-4-ylidene]-
hydrazino}-biphenyl-3- carboxylic acid 5 ##STR00007##
3'-[N'-(1-Bicyclo[4.2.0]octa- 1(6),2,4-trien-3-yl-3-methyl-5-
oxo-1,5-dihydro-pyrazol-4-ylidene)-
hydrazino]-2'-hydroxy-biphenyl-3- carboxylic acid 6 ##STR00008##
5-(2-Hydroxy-3-{N'-[3-methyl-5- oxo-1-(5,6,7,8-tetrahydro-
naphthalen-2-yl)-1,5-dihydro- pyrazol-4-ylidene]-hydrazino}-
phenyl)-1,2,4-trimethyl-1H- pyrrole-3-carboxylic acid ethyl ester 7
##STR00009## 3'-{N'-[1-(2,3-Dihydro- benzofuran-5-yl)-3-methyl-5-
oxo-1,5-dihydro-pyrazol-4- ylidene]-hydrazino}-2'-
hydroxy-biphenyl-3-carboxylic acid 8 ##STR00010##
2-(3,3-Dimethyl-indan-5-yl)-4-{[2- hydroxy-3'-(1H-tetrazol-5-yl)-
biphenyl-3-yl]-hydrazono}-5- methyl-2,4-dihydro-pyrazol-3-one 9
##STR00011## 5-{2-Hydroxy-3-[N'-(1-indan-5-yl-
3-methyl-5-oxo-1,5-dihydro- pyrazol-4-ylidene)-hydrazino]-
phenyl}-furan-2-carboxylic acid 10 ##STR00012##
4-{[2-Hydroxy-3'-(1H-tetrazol-5- yl)-biphenyl-3-yl]-hydrazono}-5-
methyl-2-(5,6,7,8-tetrahydro- naphthalen-2-yl)-2,4-dihydro-
pyrazol-3-one 11 ##STR00013## 2'-Hydroxy-5'-methyl-3'-{N'-[3-
methyl-5-oxo-1-(5,6,7,8- tetrahydro-naphthalen-2-yl)-1,5-
dihydro-pyrazol-4-ylidene]- hydrazino}-biphenyl-3- carboxylic acid
12 ##STR00014## 5-(3-{N'-[1-(3,3-Dimethyl-indan-5-
yl)-3-methyl-5-oxo-1,5-dihydro- pyrazol-4-ylidene]-hydrazino}-2-
hydroxy-phenyl)-thiophene-2- carboxylic acid 13 ##STR00015##
3'-{N'-[1-(2,3-Dihydro- benzofuran-5-yl)-3-methyl-5-
oxo-1,5-dihydro-pyrazol-4- ylidene]-hydrazino}-2'-
hydroxy-5'-methyl-biphenyl-3- carboxylic acid 14 ##STR00016##
3'-{N'-[1-(2,3-Dihydro-benzofuran-
5-yl)-3-methyl-5-oxo-1,5-dihydro- pyrazol-4-ylidene]-hydrazino}-5'-
fluoro-2'-hydroxy-biphenyl-3- carboxylic acid 15 ##STR00017##
5-(2-Hydroxy-3-{N'-[3-methyl-5- oxo-1-(5,6,7,8-tetrahydro-
naphthalen-2-yl)-1,5-dihydro- pyrazol-4-ylidene]-hydrazino}-
phenyl)-furan-2-carboxylic acid 16 ##STR00018##
3'-{N'-[1-(3,3-Dimethyl-indan-5- yl)-3-methyl-5-oxo-1,5-dihydro-
pyrazol-4-ylidene]-hydrazino}-2'- hydroxy-5'-methyl-biphenyl-3-
carboxyiic acid 17 ##STR00019## 3'-{N'-[1-(3,3-Dimethyl-indan-5-
yl)-3-methyl-5-oxo-1,5-dihydro- pyrazol-4-ylidene]-hydrazino}-5'-
fluoro-2'-hydroxy-biphenyl-3- carboxylic acid 18 ##STR00020##
5-{2-Hydroxy-3-[N'-(1-indan-5-yl- 3-methyl-5-oxo-1,5-dihydro-
pyrazol-4-ylidene)-hydrazino]- phenyl}-thiophene-2-carboxylic acid
19 ##STR00021## 2'-Hydroxy-3'-{N'-[3-methyl-5-
oxo-1-(1,1,3,3-tetramethyl-indan-5-
yl)-1,5-dihydro-pyrazol-4-ylidene]- hydrazino}-biphenyl-3-
carboxylic acid 20 ##STR00022## 2'-Hydroxy-5'-methyl-3'-{N'-[3-
methyl-5-oxo-1-(1,1,3,3- tetramethyl-indan-5-yl)-1,5-
dihydro-pyrazol-4-ylidene]- hydrazino}-biphenyl-3- carboxylic acid
21 ##STR00023## 3'-[N'-(1-Bicyclo[4.2.0]octa-
1(6),2,4-trien-3-yl-3-methyl-5- oxo-1,5-dihydro-pyrazol-4-
ylidene)-hydrazino]-2'-hydroxy- 5'-methyl-biphenyl-3- carboxylic
acid 22 ##STR00024## 5-{3-[N'-(1-Bicyclo[4.2.0]octa-
1(6),2,4-trien-3-yl-3-methyl-5-oxo- 1,5-dihydro-pyrazol-4-ylidene)-
hydrazino]-2-hydroxy-phenyl}- furan-2-carboxylic acid 23
##STR00025## 2-Bicyclo[4.2.0]octa-1(6),2,4-trien-
3-yl-4-{[2-hydroxy-3'-(1H-tetrazol-
5-yl)-biphenyl-3-yl]-hydrazono}-5- methyl-2,4-dihydro-pyrazol-3-one
24 ##STR00026## 5-{2-Hydroxy-3-[N'-(1-indan-5-yl-
3-methyl-5-oxo-1,5-dihydro- pyrazol-4-ylidene)-hydrazino]-5-
methyl-phenyl}-thiophene-2- carboxylic acid 25 ##STR00027##
5-(2-Hydroxy-3-{N'-[3-methyl-5- oxo-1-(1,1,3,3-tetramethyl-indan-5-
yl)-1,5-dihydro-pyrazol-4-ylidene]- hydrazino}-phenyl)-furan-2-
carboxylic acid 26 ##STR00028## 3'-[N'-(1-BicycIo[4.2.0]octa-
1(6),2,4-trien-3-yl-3-methyl-5- oxo-1,5-dihydro-pyrazol-4-ylidene)-
hydrazino]-5'-fluoro-2'-hydroxy- biphenyl-3-carboxylic acid 27
##STR00029## 4-{[2-Hydroxy-3'-(1H-tetrazol-5-
yl)-biphenyl-3-yl]-hydrazono}-2- indan-5-yl-5-methyl-2,4-dihydro-
pyrazol-3-one 28 ##STR00030## 4-(2-Hydroxy-3-{N'-[3-methyl-5-
oxo-1-(5,6,7,8-tetrahydro- naphthalen-2-yl)-1,5-dihydro-
pyrazol-4-ylidene]-hydrazino}- phenyl)-furan-2-carboxylic acid 29
##STR00031## 2'-Hydroxy-3'-[N'-(1-indan-5-yl-3-
methyl-5-oxo-1,5-dihydro-pyrazol- 4-ylidene)-hydrazino]-5'-methyl-
biphenyl-3-carboxylic acid 30 ##STR00032##
3'-{N'-[1-(3,3-Dimethyl-indan-5- yl)-3-methyl-5-oxo-1,5-dihydro-
pyrazol-4-ylidene]-hydrazino}- 2'-hydroxy-biphenyl-3- carboxylic
acid 31 ##STR00033## 4-{2-Hydroxy-3-[N'-(1-indan-5-yl-
3-methyl-5-oxo-1,5-dihydro- pyrazol-4-ylidene)-hydrazino]-
phenyl}-furan-2-carboxylic acid 32 ##STR00034##
4-{[4'-(4,5-Dihydro-1H-imidazol- 2-yl)-2-hydroxy-biphenyl-3-yl]-
hydrazono}-2-indan-5-yl-5-methyl- 2,4-dihydro-pyrazol-3-one 33
##STR00035## 5-(2-Hydroxy-5-methyl-3-{N'-[3-
methyl-5-oxo-1-(5,6,7,8- tetrahydro-naphthalen-2-yl)-1,5-
dihydro-pyrazol-4-ylidene]- hydrazino}-phenyl)-thiophene-
2-carboxylic acid 34 ##STR00036##
5-(3-{N'-[1-(3,3-Dimethyl-indan-5- yl)-3-methyl-5-oxo-1,5-dihydro-
pyrazol-4-ylidene]-hydrazino}-2- hydroxy-5-methyl-phenyl)-
thiophene-2-carboxylic acid 35 ##STR00037##
5-{3-[N'-(1-Bicyclo[4.2.0]octa- 1(6),2,4-trien-3-yl-3-methyl-5-oxo-
1,5-dihydro-pyrazol-4-ylidene)- hydrazino]-2-hydroxy-5-methyl-
phenyl}-thiophene-2-carboxylic acid 36 ##STR00038##
5-(2-Hydroxy-3-{N'-[3-methyl-5- oxo-1-(5,6,7,8-tetrahydro-
naphthalen-2-yl)-1,5-dihydro- pyrazol-4-ylidene]-hydrazino}-
phenyl)-thiophene-2-carboxylic acid 37 ##STR00039##
5-{3-[N'-(1-Bicyclo[4.2.0]octa- 1(6),2,4-trien-3-yl-3-methyl-5-
oxo-1,5-dihydro-pyrazol-4- ylidene)-hydrazino]-2-hydroxy-
phenyl}-thiophene-2-carboxylic acid 38 ##STR00040##
2'-Hydroxy-3'-{N'-[3-methyl-1-(3- methyl-indan-5-yl)-5-oxo-1,5-
dihydro-pyrazol-4-ylidene]- hydrazino}-biphenyl-3-carboxylic acid
39 ##STR00041## 2'-Hydroxy-5'-methyl-3'-{N'-[3-
methyl-1-(3-methyl-indan-5-yl)-5- oxo-1,5-dihydro-pyrazol-4-
ylidene]-hydrazino}-biphenyl-3- carboxylic acid 40 ##STR00042##
5-(2-Hydroxy-3-{N'-[3-methyl-1- (3-methyl-indan-5-yl)-5-oxo-1,5-
dihydro-pyrazol-4-ylidene]- hydrazino}-phenyl)-thiophene-2-
carboxylic acid 41 ##STR00043## 3'-{N'-[1-(3-Ethyl-indan-5-yl)-3-
methyl-5-oxo-1,5-dihydro-pyrazol- 4-ylidene]-hydrazino}-2'-hydroxy-
biphenyl-3-carboxylic acid 42 ##STR00044##
3'-{N'-[1-(3-Ethyl-indan-5-yl)-3- methyl-5-oxo-1,5-dihydro-pyrazol-
4-ylidene]-hydrazino}-2'-hydroxy- 5'-methyl-biphenyl-3-carboxylic
acid 43 ##STR00045## 5-(3-{N'-[1-(3,3-Dimethyl-indan-5-
yl)-3-methyl-5-oxo-1,5-dihydro- pyrazol-4-ylidene]-hydrazino}-2-
hydroxy-phenyl)-furan-2- carboxylic acid 44 ##STR00046##
5-(2-Hydroxy-3-{N'-[3-methyl-1- (3-methyl-indan-5-yl)-5-oxo-1,5-
dihydro-pyrazol-4-ylidene]- hydrazino}-phenyl)-furan-2- carboxylic
acid 45 ##STR00047## 5-(3-{N'-[1-(2,2-Dimethyl-indan-5-
yl)-3-methyl-5-oxo-1,5-dihydro- pyrazol-4-ylidene]-hydrazino}-2-
hydroxy-phenyl)-furan-2- carboxylic acid 46 ##STR00048##
5-{2-Hydroxy-3-[N'-(1-indan-5-yl- 3-methyl-5-oxo-1,5-dihydro-
pyrazol-4-ylidene)-hydrazino]-5- methyl-phenyl}-furan-2-carboxylic
acid 47 ##STR00049## 2-{2-Hydroxy-3-[N'-(1-indan-5-yl-
3-methyl-5-oxo-1,5-dihydro- pyrazol-4-ylidene)-hydrazino]-
phenyl}-5-methyl-thiazole-4- carboxylic acid 48 ##STR00050##
5-{2-Hydroxy-3-[N'-(1-indan-5-yl- 3-methyl-5-oxo-1,5-dihydro-
pyrazol-4-ylidene)-hydrazino]- phenyl}-1,2,4-trimethyl-1H-
pyrrole-3-carboxylic acid ethyl ester 49 ##STR00051##
5-(2-Hydroxy-3-{N'-[3-methyl-5- oxo-1-(1,1,3,3-tetramethyl-indan-5-
yl)-1,5-dihydro-pyrazol-4-ylidene]- hydrazino}-phenyl)-thiophene-2-
carboxylic acid 50 ##STR00052## 5-(2-Hydroxy-5-methyl-3-{N'-[3-
methyl-5-oxo-1-(5,6,7,8- tetrahydro-naphthalen-2-yl)-1,5-
dihydro-pyrazol-4-ylidene]- hydrazino}-phenyl)-furan-2- carboxylic
acid 51 ##STR00053## 4-(2-Hydroxy-3-{N'-[3-methyl-5-
oxo-1-(5,6,7,8-tetrahydro- naphthalen-2-yl)-1,5-dihydro-
pyrazol-4-ylidene]-hydrazino}- phenyl)-thiophene-2-carboxylic acid
52 ##STR00054## 4-{2-Hydroxy-3-[N'-(1-indan-5-yl-
3-methyl-5-oxo-1,5-dihydro- pyrazol-4-ylidene)-hydrazino]-
phenyl}-thiophene-2-carboxylic acid 53 ##STR00055##
5-(2-Hydroxy-3-{N'-[3-methyl-5- oxo-1-(5,6,7,8-tetrahydro-
naphthalen-2-yl)-1,5-dihydro- pyrazol-4-ylidene]-hydrazino}-
phenyl)-2-methyl-furan-3- carboxylic acid 54 ##STR00056##
5-(2-Hydroxy-3-{N'-[3-methyl-5- oxo-1-(5,6,7,8-tetrahydro-
naphthalen-2-yl)-1,5-dihydro- pyrazol-4-ylidene]-hydrazino}-
phenyl)-furan-2-carboxylic acid methyl ester 55 ##STR00057##
5-{2-Hydroxy-3-[N'-(1-indan-5-yl- 3-methyl-5-oxo-1,5-dihydro-
pyrazol-4-ylidene)-hydrazino]- phenyl}-furan-2-carboxylic acid
methyl ester 56 ##STR00058## 3'-{N'-[1-(2,2-Dimethyl-indan-5-
yl)-3-methyl-5-oxo-1,5-dihydro- pyrazol-4-ylidene]-hydrazino}-2'-
hydroxy-biphenyl-3-carboxylic acid 57 ##STR00059##
4-(2-Hydroxy-3-{N'-[3-methyl-5- oxo-1-(5,6,7,8-tetrahydro-
naphthalen-2-yl)-1,5-dihydro- pyrazol-4-ylidene]-hydrazino}-
phenyl)-1H-pyrrole-2-carboxylic acid 58 ##STR00060##
4-{2-Hydroxy-3-[N'-(1-indan-5-yl- 3-methvl-5-oxo-1,5-dihydro-
pyrazol-4-ylidene)-hydrazino]- phenyl}-1H-pyrrole-2-carboxylic
acid
or pharmaceutical acceptable salts, hydrates or solvates
thereof.
[0026] The present disclosure further provides the compounds of
formula (IA), as intermediates in the synthesis of the compounds
having formula (I):
##STR00061##
[0027] wherein:
[0028] A is selected from the group consisting of carbon and
oxygen;
[0029] R.sup.5, R.sup.6 and R.sup.7 are each independently selected
from the group consisting of hydrogen, alkyl, alkoxy, halogen,
hydroxyl, amino, nitro, cyano, carboxylic acid and carboxylic
ester;
[0030] R.sup.8, R.sup.9, R.sup.10 and R.sup.11 are each
independently selected from the group consisting of hydrogen and
alkyl; and
[0031] n is 0, 1 or 2.
[0032] The compounds of formula (IA) of the present disclosure
preferably include, but are not limited to:
TABLE-US-00002 No. Structure Name 1 ##STR00062##
2-Bicyclo[4.2.0]octa- 1(6),2,4-trien-3-yl-5- methyl-2,4-dihydro-
pyrazol-3-one 2 ##STR00063## 2-Indan-5-yl-5-methyl-
2,4-dihydro-pyrazol- 3-one 3 ##STR00064## 5-Methyl-2-(5,6,7,8-
tetrahydro-naphthalen- 2-yl)-2,4-dihydro- pyrazol-3-one 4
##STR00065## 2-(2,3-Dihydro- benzofuran-5-yl)-5-
methyl-2,4-dihydro- pyrazol-3-one 5 ##STR00066##
5-Methyl-2-(3-methyl- indan-5-yl)-2,4- dihydro-pyrazol- 3-one 6
##STR00067## 2-(3-Ethyl-indan-5-yl)- 5-methyl-2,4-
dihydro-pyrazol-3- one 7 ##STR00068## 2-(3,3-Dimethyl-indan-
5-yl)-5-methyl-2,4- dihydro-pyrazol- 3-one 8 ##STR00069##
2-(2,2-Dimethyl-indan- 5-yl)-5-methyl-2,4- dihydro-pyrazol- 3-one 9
##STR00070## 5-Methyl-2-(1,1,3,3- tetramethyl-indan-5-
yl)-2,4-dihydro- pyrazol-3-one
[0033] In another embodiment, the present disclosure provides a
process for the preparation of the compounds of formula (IA),
comprising the following steps of =
##STR00071##
[0034] reacting an amino substituted benzocycle and sodium nitrite
in an acidic solution via a diazo reaction; reducing the resulting
intermediate by tin dichloride to obtain a hydrazine; heating the
hydrazine and an electrophilic carbonyl compound, such as ethyl
acetoacetate, in an suitable solvent, such as acetic acid, ethanol
and the like, via a coupling reaction to obtain the compound of
formula (IA).
[0035] In a further embodiment, the present disclosure provides a
process for the preparation of the compounds of formula (I),
comprising the following steps of:
##STR00072##
[0036] reacting a substituted aniline compound and sodium nitrite
in a suitable acid, such as nitric acid, sulfuric acid,
hydrochloric acid, via a diazo-reaction; reacting the resulting
intermediate and the compound of formula (IA) in a suitable base,
such as sodium bicarbonate, potassium hydrogen carbonate, via a
coupling reaction to obtain the compound of formula (I).
[0037] The present disclosure relates to a use of the compounds of
formula (I) and formula (IA) in the preparation of TPO receptor
agonists.
[0038] The present disclosure relates to a use of the compounds of
formula (I) and formula (IA) in the preparation of a medicament for
the treatment of thrombocytopenia. Further, the said medicament can
be co-administered with a therapeutically effective amount of one
or more drugs selected from the group consisting of a colony
stimulating factor, a cytokine, a chemokine, an interleukin or
cytokine receptor agonist or antagonist, a soluble receptor, a
receptor agonist or antagonist antibody, or one or more peptides or
small molecule compounds which have the same mechanism of the said
drugs.
[0039] The present disclosure relates to a pharmaceutical
composition comprising a therapeutically effective amount of the
compounds of formula (I) and formula (IA), or pharmaceutically
acceptable salts, hydrates or solvates thereof. Further, the said
composition can be co-administered with a therapeutically effective
amount of one or more drugs selected from the group consisting of a
colony stimulating factor, a cytokine, a chemokine, an interleukin
or cytokine receptor agonist. The present disclosure also relates
to a use of the said pharmaceutical composition in the preparation
of a medicament for the treatment of thrombocytopenia.
[0040] The term "co-administering" means either simultaneous
administration or separate sequential administration of the
compounds of the present disclosure.
[0041] The present disclosure relates to a process for the
preparation of a pharmaceutical composition comprising a
pharmaceutically acceptable carrier or diluent and a
therapeutically effective amount of the compounds of formula (I)
and formula (IA), as well as their pharmaceutically acceptable
salts, hydrates or solvates, wherein the process comprises
combining the compounds of formula (I) and formula (IA) with
carriers and diluents.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0042] Unless otherwise stated, the following terms used in the
specification and claims have the meanings defined as below.
[0043] The term "alkyl" refers to a saturated aliphatic radical
including straight chain or branched chain hydrocarbon radical
having 1 to 20 carbon atoms. The alkyl group is preferably an alkyl
having 1 to 10 carbon atoms, such as methyl, ethyl, propyl,
2-propyl, n-butyl, iso-butyl, tert-butyl or pentyl and the like.
The alkyl group is more preferably a lower alkyl having 1 to 4
carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl,
iso-butyl or tert-butyl and the like. The alkyl group may be
substituted or unsubstituted. When substituted, the substituent is
preferably one or more groups independently selected from the group
consisting of alkyl, alkoxy, halogen, hydroxyl, amino, nitro,
cyano, aryl, heteroaryl, carboxylic acid or carboxylic ester.
[0044] The term "aryl" refers to a radical having at least one
aromatic ring, i.e. having a conjugated pi-electron system,
including all-carbon cyclic aryl, heteroaryl and biaryl. The aryl
group may be substituted or unsubstituted. When substituted, the
substituent is preferably one or more groups independently selected
from the group consisting of alkyl, alkoxy, halogen, hydroxyl,
amino, nitro, cyano, aryl, heteroaryl, carboxylic acid or
carboxylic ester.
[0045] The term "heteroaryl" refers to an aryl having 1 to 4
heteroatoms selected from the group consisting of N, O, and S as
ring atoms, the remaining ring atoms being C. The said ring may be
a 5- or 6-membered ring. Examples of heteroaryl groups include
furyl, thienyl, pyridyl, pyrrolyl, N-alkyl pyrrolyl, pyrimidinyl,
pyrazinyl, imidazolyl, tetrazolyl, and the like. The heteroaryl
group may be substituted or unsubstituted. When substituted, the
substituent is preferably one or more groups independently selected
from the group consisting of alkyl, alkoxy, halogen, hydroxyl,
amino, nitro, cyano, aryl, heteroaryl, carboxylic acid or
carboxylic ester.
[0046] The term "hydroxy" refers to an --OH radical.
[0047] The term "alkoxyl" refers to both an --O-(alkyl) and an
--O-(unsubstituted cycloalkyl) radical, Representative examples
include, but are not limited to, methoxy, ethoxy, propoxy, butoxy,
cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and
the like. The alkoxyl group may be substituted or unsubstituted.
When substituted, the substituent is preferably one or more groups
independently selected from the group consisting of alkyl, alkoxy,
halogen, hydroxyl, amino, nitro, cyano, aryl, heteroaryl,
carboxylic acid or carboxylic ester.
[0048] The term "halogen" refers to fluoro, chloro, bromo or
iodo.
[0049] The term "amino" refers to a --NH.sub.2 radical.
[0050] The term "cyano" refers to a --CN radical.
[0051] The term "nitro" refers to a --NO.sub.2 radical.
[0052] The term "carboxylic acid" refers to a (alkyl) C(.dbd.O)OH
radical.
[0053] The term "carboxylic ester" refers to a (alkyl) C(.dbd.O)O
(alkyl).
[0054] The term "pharmaceutical composition" refers to a mixture of
one or more of the compounds described herein, or
physiologically/pharmaceutically acceptable salts or prodrugs
thereof, with other chemical components such as
physiologically/pharmaceutically acceptable carriers and
excipients. The purpose of a pharmaceutical composition is to
facilitate administration of a compound to an organism.
Synthesis Methods of the Disclosed Compounds
##STR00073##
[0056] reacting an amino substituted benzocycle and sodium nitrite
in an acidic solution via a diazo-reaction; reducing the resulting
intermediate by tin dichloride to obtain a hydrazine; heating the
hydrazine and an electrophilic carbonyl compound, such as ethyl
acetoacetate, in a suitable solvent, such as acetic acid, ethanol
and the like, via a coupling reaction to obtain the compound of
formula (IA).
##STR00074##
[0057] nitrating a substituted 2-bromophenol by sodium nitrate to
obtain a nitrophenol, which is converted to a hydroxyl protected
nitrophenol via a alkylation reaction at a suitable temperature, in
the presence of a haloalkyl, such as methyl iodide; reacting the
hydroxyl protected nitrophenol and a substituted arylboronic acid
via a Suzuki coupling reaction in the presence of the catalyst
tetrakis(triphenylphosphine)palladium; or reacting the hydroxyl
protected nitrophenol and a boronic acid derivative to obtain an
arylboronic acid compound, which is reacted with a halogenated
compound R.sup.1X via a Suzuki coupling reaction to obtain a
R.sup.1 substituted aryl compound; reducing the R.sup.1 substituted
aryl compound by palladium on carbon under hydrogen atmosphere to
obtain arylaniline; removing the alkyl protecting group in the
presence of hydrobromic acid to obtain the unprotected aniline; or
removing the alkyl protecting group of the substituted aryl
compound in the presence of hydrobromic acid to obtain a nitro
compound, which is reduced by palladium on carbon under hydrogen
atmosphere to obtain the unprotected arylaniline.
[0058] Reacting the substituted aniline compound and sodium nitrite
in a suitable acid, such as nitric acid, sulfuric acid,
hydrochloric acid, via a diazo-reaction; reacting the resulting
intermediate and the compound of formula (IA) in a suitable base,
such as sodium bicarbonate, potassium hydrogen carbonate, via a
coupling reaction to obtain the compound of formula (I).
[0059] The present disclosure is further described by the following
examples which are not intended to limit the scope of the
disclosure.
EXAMPLES
[0060] The structures of all compounds were identified by nuclear
magnetic resonance (.sup.1H NMR) or mass spectrometry (MS). .sup.1H
NMR chemical shifts were recorded on ppm (10.sup.-6). .sup.1H NMR
was performed on a Bruker AVANCE-400 spectrometer. The appropriate
solvents were deuterated-methanol (CD.sub.3OD),
deuterated-chloroform (CDCl.sub.3) and deuterated-dimethyl
sulfoxide (DMSO-d.sub.6) with tetramethylsilane (TMS) as the
internal standard and chemical shifts were recorded on ppm
(10.sup.-6).
[0061] MS was determined by a FINNIGAN LCQ Ad (ESI) mass
spectrometer (Thermo, Model: Finnigan LCQ advantage MAX).
[0062] IC.sub.50 was determined by a NovoStar ELIASA (BMG Co.
German).
[0063] The type of thin-layer silica, gel was Yantai Huanghai
HSGF254 or Qingdao GF254 silica gel plate.
[0064] Column chromatography studies generally used Yantai Huanghai
200.about.300 mesh silica gel as carrier.
[0065] HPLC was determined by an Agilent 1200DAD high pressure
liquid chromatography spectrometer (Sunfire C18 150.times.4.6 mm
chromatographic column) and a Waters 2695-2996 high pressure liquid
chromatography spectrometer (Gimini C18 150.times.4.6 mm
chromatographic column).
[0066] Hydrogenation reactions under pressure were performed with a
Pau 3916EKX hydrogenation spectrometer and a QL hydrogen generator.
Microwave reactions were performed with a CEM Discover-S 908860
microwave reactor.
[0067] Unless otherwise stated, the following reactions were placed
under nitrogen atmosphere.
[0068] The term "nitrogen atmosphere" refers to that a reaction
flask is equipped with an about 1 L nitrogen balloon.
[0069] The term "hydrogen atmosphere" refers to that a reaction
flask is equipped with an about 1 L hydrogen balloon.
[0070] Unless otherwise stated, the solution used in following
reaction refers to an aqueous solution.
[0071] The term "TLC" refers to thin layer chromatography.
Example 1
2'-Hydroxy-3'-N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-yliden-
e)-hydrazino-biphenyl-3-carboxylic acid
##STR00075## ##STR00076##
[0072] Step 1
2-Bromo-6-nitro-phenol
[0073] A solution of 60 mL of concentrated sulfuric acid diluted
with 186 mL of water was cooled to room temperature. Sodium nitrate
(79.2 g, 0.932 mol) was added to the solution. 2-Bromo-phenol 1a
(60 mL, 0.516 mmol) was added dropwise at such a rate that the
reaction temperature was kept below 25.degree. C. The reaction
mixture was reacted at room temperature for 2 hours and monitored
by thin layer chromatography (TLC) until the disappearance of the
starting materials. The precipitate was dissolved in 320 mL of
ethyl acetate. The mixture was washed with saturated brine, dried
over anhydrous magnesium sulfate, filtered and concentrated under
reduced pressure. The resulting residue was purified by silica gel
column chromatography to obtain the title compound
2-bromo-6-nitro-phenol 1b (48.2 g, yield 42.8%) as a yellow
solid.
[0074] MS m/z (ESI): 218 [M+1]
[0075] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 6.88.about.7.02
(m, 1H), 7.89.about.7.91 (d, J=8 Hz, 1H), 8.12.about.8.15 (m, 1H),
11.18 (s, 1H)
Step 2
1-Bromo-2-methoxy-3-nitro-benzene
[0076] 2-Bromo-6-nitro-phenol 1b (46.55 g, 0.214 mol) was dissolved
in 500 mL of acetone followed by addition of potassium carbonate
(35.36 g, 0.256 mol) and iodomethane (20.1 mL, 0.32 mol). The
reaction mixture was heated to reflux at 70.degree. C. for 40
hours. The reaction was monitored by TLC until the disappearance of
the starting materials. The reaction mixture was concentrated under
reduced pressure and diluted with 1300 mL of ethyl acetate and 500
mL of water. The aqueous layer was extracted with ethyl acetate
(300 mL.times.2). The combined organic extracts were washed with 4
N hydrochloric acid and saturated aqueous sodium bicarbonate and
then dried over anhydrous magnesium sulfate, filtered and
concentrated under reduced pressure. The resulting residue was
purified by silica gel column chromatography to obtain the title
compound 1-bromo-2-methoxy-3-nitro-benzene 1c (44.59 g, yield
90.0%) as a brown solid.
[0077] MS m/z (ESI): 234 [M+1]
Step 3
2'-Methoxy-3'-nitro-biphenyl-3-carboxylic acid
[0078] 1-Bromo-2-methoxy-3-nitro-benzene 1c (23.25 g, 0.10 mol),
3-carboxyphenylboronic acid (19.5 g, 0.117 mol) and
tetrakis(triphenylphosphine)palladium (8.86 g, 7.7 mol) were
dissolved in a solvent mixture of 100 mL of 2 N aqueous sodium
carbonate and 500 mL of 1,4-dioxane. The reaction mixture was
heated to reflux at 105.degree. C. for 43 hours. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was concentrated under reduced pressure and then 300 mL
of 6 N hydrochloric acid and 400 mL of ethyl acetate were added.
The aqueous layer was extracted with ethyl acetate (200
mL.times.2). The combined organic extracts were dried over
anhydrous magnesium sulfate, filtered and concentrated under
reduced pressure to obtain the title compound
2'-methoxy-3'-nitro-biphenyl-3-carboxylic acid 1d (53.93 g) as a
light yellow solid.
[0079] MS m/z (ESI): 272 [M-1]
[0080] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 3.44.about.3.46
(d, J=8 Hz, 3H), 7.42.about.7.46 (m, 1H), 7.63.about.7.67 (m, 1H),
7.21.about.7.75 (m, 1H), 7.82.about.7.84 (m, 1H), 7.90.about.7.92
(m, 1H), 8.01.about.8.03 (d, J=8 Hz, 1H), 8.11 (s, 1H)
Step 4
2'-Methoxy-3'-amino-biphenyl-3-carboxylic acid
[0081] 2'-Methoxy-3'-nitro-biphenyl-3-carboxylic acid 1d (0.48 g,
1.74 mmol) was dissolved in 60 mL of ethanol followed by addition
of 0.5 g of palladium on carbon and ammonium formate (1.1 g, 17.4
mmol). The reaction mixture was heated to reflux at 80.degree. C.
for 20 minutes. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filtrate was concentrated under reduced pressure and dried
to obtain the title compound
2'-methoxy-3'-amino-biphenyl-3-carboxylic acid 1e (0.42 g, yield
93.3%) as a white solid.
[0082] MS m/z (ESI): 242 [M-1]
Step 5
3'-Amino-2'-hydroxy-biphenyl-3-carboxylic acid hydrobromide
[0083] 2'-Methoxy-3'-amino-biphenyl-3-carboxylic acid 1e (2.5 g,
10.3 mmol) was dissolved in 100 mL of hydrobromic acid (40%). The
reaction mixture was heated to reflux at 120.degree. C. overnight,
and the reaction was monitored by TLC until the disappearance of
the starting materials. The mixture was concentrated under reduced
pressure and the resulting residue was purified by silica gel
column chromatography to obtain the title compound
3'-amino-2'-hydroxy-biphenyl-3-carboxylic acid hydrobromide 1f (2.4
g, 88.8%) as a khaki solid.
[0084] MS m/z (ESI): 230 [M+1]
[0085] [Reference: WO01/89457]
Step 6
Indan-5-yl-hydrazine
[0086] Indan-5-ylamine 1g (3.59 g, 27.0 mmol) was dissolved in 20
mL of concentrated hydrochloric acid upon cooling by an ice-water
bath and the mixture was stirred for 10 minutes. 10 mL of aqueous
sodium nitrite (1.86 g, 27.0 mmol) was added dropwise and the
mixture was stirred for another 15 minutes and used in the
following reaction.
[0087] Upon cooling by an ice-salt bath, stannous chloride
dihydrate (24.4 g, 108.0 mmol) was dissolved in 10 mL of
concentrated hydrochloric acid followed by addition of above
mentioned spare mixture. The reaction mixture was warmed up to room
temperature and reacted for 1.5 hours. Then the mixture was
adjusted to pH 9 with 40% aqueous sodium hydroxide upon cooling by
an ice-water bath. The mixture was extracted with 400 mL of ethyl
acetate and the combined organic extracts were concentrated under
reduced pressure and dried to obtain the title compound
indan-5-yl-hydrazine 1h (2.05 g, yield 51.3%) as a rufous
solid.
[0088] MS m/z (ESI): 149 [M+1]
Step 7
2-Indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one
[0089] Indan-5-yl-hydrazine 1h (2.05 g, 13.8 mmol) was dissolved in
50 mL of acetic acid followed by addition of ethyl acetoacetate
(1.76 mL, 13.8 mmol). The reaction mixture was heated at
100.degree. C. overnight and the reaction was monitored by TLC
until the disappearance of the starting materials. The mixture was
concentrated under reduced pressure and the resulting residue was
purified by silica gel column chromatography to obtain the title
compound 2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (1.84
g, yield 62.3%) as a yellow solid.
[0090] MS m/z (ESI): 215 [M+1]
[0091] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.69 (s, 1H),
7.60 (d, J=8.0 Hz, 1H), 7.24 (d, J=8 Hz, 1H), 3.44 (s, 2H),
2.90.about.2.97 (m, 4H), 3.21 (s, 3H), 2.07.about.2.14 (m, 2H)
Step 8
2'-Hydroxy-3'-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylide-
ne)-hydrazino]-biphenyl-3-carboxylic acid
[0092] Upon cooling by an ice-salt bath,
3'-amino-2'-hydroxy-biphenyl-3-carboxylic acid hydrobromide 1f (267
mg, 1.16 mmol) was dissolved in 10 mL of hydrochloric acid (1 N)
followed by addition of 10 mL of aqueous sodium nitrite (88 mg,
1.28 mmol) and 2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i
(249 mg, 1.16 mmol). The mixture was adjusted to pH 8 with
saturated aqueous sodium bicarbonate, followed by addition of 10 mL
of ethanol. The reaction mixture was warmed up to room temperature
overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered,
dried and recrystallized from methanol to obtain the title compound
2'-hydroxy-3'-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylid-
ene)-hydrazino]-biphenyl-3-carboxylic acid 1 (60 mg, yield 11.4%)
as a yellow solid.
[0093] MS m/z (ESI): 453 [M-1]
[0094] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.03.about.2.10
(m, 2H), 2.34 (s, 3H), 2.86.about.2.93 (m, 4H), 7.13.about.7.17 (m,
2H), 7.28.about.7.30 (d, J=8.1 Hz, 1H), 7.60.about.7.82 (m, 5H),
7.96.about.7.98 (d, J=8.1 Hz, 1H), 8.13 (s, 1H), 9.66 (s, 1H),
13.03 (s, 1H), 13.76 (s, 1H)
Example 2
5'-Fluoro-2'-hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazo-
l-4-ylidene)-hydrazino]-biphenyl-3-carboxylic acid
##STR00077## ##STR00078##
[0095] Step 1
2-Bromo-4-fluoro-6-nitro-phenol
[0096] 2-Bromo-4-fluoro-phenol 2a (8.0 g, 41.9 mmol) was dissolved
in 10 mL of sulfuric acid (50%) followed by addition of a solution
of sodium nitrate (7.1 g, 83.5 mmol) in 24 mL of sulfuric acid
(25%), upon cooling by an ice-water bath. The reaction mixture was
reacted at room temperature for 1.5 hours and the reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was diluted with 50 mL of water and extracted with
ethyl acetate (50 mL.times.2). The combined organic extracts were
washed with water and saturated aqueous sodium bicarbonate and
dried over anhydrous magnesium sulfate, filtered and concentrated
under reduced pressure to obtain the title compound
2-bromo-4-fluoro-6-nitro-phenol 2b (8.0 g, yield 80.8%) as a red
solid, which was directly used in the next step.
Step 2
1-Bromo-5-fluoro-2-methoxy-3-nitro-benzene
[0097] 2-Bromo-4-fluoro-6-nitro-phenol 2b (24.7 g, 104.7 mmol) and
potassium carbonate (17.34 g, 125.6 mmol) were dissolved in 300 mL
of acetone followed by addition of iodomethane (9.8 mL, 157.1
mmol). The reaction mixture was heated to reflux at 80.degree. C.
for 22 hours. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was
concentrated under reduced pressure and diluted with 200 mL of
ethyl acetate and 200 mL of water. The aqueous layer was extracted
with ethyl acetate (100 mL.times.2). The combined organic extracts
were washed with 4 N hydrochloric acid and saturated aqueous sodium
bicarbonate and dried over anhydrous magnesium sulfate, filtered
and concentrated under reduced pressure. The resulting residue was
purified by silica gel column chromatography to obtain the title
compound 1-bromo-5-fluoro-2-methoxy-3-nitro-benzene 2c (16.18 g,
yield 61.8%) as a white solid.
[0098] MS m/z (ESI): 252 [M+1]
[0099] .sup.1H NMR (CDCl.sub.3): .delta. 3.99 (s, 3H), 7.81 (d,
J=8.0 Hz, 1H), 7.28 (q, J=8.0 Hz, 4.0 Hz, 1H), 7.89 (q, J=8.0 Hz,
4.0 Hz, 1H)
Step 3
5'-Fluoro-2'-methoxy-3'-nitro-biphenyl-3-carboxylic acid
[0100] 1-Bromo-5-fluoro-2-methoxy-3-nitro-benzene 2c (16.18 g, 64.7
mmol), 3-carboxyphenylboronic acid (13.88 g, 77.7 mmol) and
tetrakis(triphenylphosphine)palladium (3.73 g, 3.2 mmol) were
dissolved in the solvent mixture of 65 mL of aqueous sodium
carbonate (2 N) and 300 mL of 1,4-dioxane. The reaction mixture was
heated to reflux at 120.degree. C. for 24 hours. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was concentrated under reduced pressure and diluted
with 150 mL of hydrochloric acid (6 N) and 200 mL of ethyl acetate.
The aqueous layer was extracted with ethyl acetate (100
mL.times.2). The combined organic extracts were dried over
anhydrous magnesium sulfate, filtered and concentrated under
reduced pressure to obtain the title compound
5'-fluoro-2'-methoxy-3'-nitro-biphenyl-3-carboxylic acid 2d (7.86
g, yield 41.7%) as a yellow solid.
[0101] MS m/z (ESI): 290 [M-1]
Step 4
[0102] 3'-Nitro-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic acid
[0103] 5'-Fluoro-2'-methoxy-3'-nitro-biphenyl-3-carboxylic acid 2d
(2.91 g, 10.0 mmol) was dissolved in 10 mL of hydrobromic acid
(40%). The reaction mixture was heated to reflux at 120.degree. C.
overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was
concentrated under reduced pressure and the resulting residue was
purified by silica gel column chromatography to obtain the title
compound 3'-nitro-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic acid
2e (2.38 g, yield 85.7%) as a yellow solid, which was directly used
in the next step.
[0104] MS m/z (ESI): 277 [M-1]
Step 5
3'-Amino-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic acid
[0105] 3'-Nitro-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic acid
hydrobromide 2e (417 mg, 1.5 mmol) was dissolved in 60 mL of
ethanol followed by addition of 0.5 g of palladium on carbon and
ammonium formate (0.95 g, 1.5 mmol). The reaction mixture was
heated to reflux at 80.degree. C. for 20 minutes. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered and the filtrate was concentrated under
reduced pressure and dried to obtain the title compound
3'-amino-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic acid 2f (339
mg, yield 91.5%) as a purple solid.
[0106] MS m/z (ESI): 246 [M-1]
Step 6
5'-Fluoro-2'-hydroxy-3'-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyraz-
ol-4-ylidene)-hydrazino]-biphenyl-3-carboxylic acid
[0107] Upon cooling by an ice-water bath,
3'-amino-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic acid 2f (296
mg, 1.20 mmol) was dissolved in 10 mL of hydrochloric acid (1 N)
followed by addition of 10 mL of aqueous sodium nitrite (91 mg,
1.32 mmol) and 2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i
(257 mg, 1.20 mmol). The mixture was adjusted to pH 8 with
saturated aqueous sodium bicarbonate, followed by addition of 10 mL
of ethanol. The mixture was warmed up to room temperature
overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered,
dried and recrystallized from methanol to obtain the title compound
5'-fluoro-2'-hydroxy-3'-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dih-
ydro-pyrazol-4-ylidene)-hydrazino]-biphenyl-3-carboxylic acid 2 (87
mg, yield 14.1%) as a red solid.
[0108] MS m/z (ESI): 471 [M-1]
[0109] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.02 (m, 2H),
2.34 (s, 3H), 2.87 (m, 4H), 7.03 (dd, J.sub.1=9.2 Hz, J.sub.2=2.8
Hz, 1H), 7.28 (d, J=8.0 Hz, 1H), 7.48 (m, 1H), 7.61 (m, 2H), 7.76
(s, 1H), 7.83 (d, J=7.6 Hz, 1H), 7.98 (d, J=8.0 Hz, 1H), 8.17 (s,
1H), 9.59 (s, 1H), 13.03 (s, 1H), 13.62 (s, 1H)
Example 3
2'-Hydroxy-3'-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)-1-
,5-dihydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic
acid
##STR00079##
[0110] Step 1
(5,6,7,8-Tetrahydro-naphthalen-2-yl)-hydrazine
[0111] 5,6,7,8-Tetra-hydro-naphthalen-2-ylamine 3g (3.68 g, 25.0
mmol) was dissolved in 20 mL of concentrated hydrochloric acid and
the mixture was stirred for minutes upon cooling by an ice-water
bath. 10 mL of aqueous sodium nitrite (1.72 g, 25.0 mmol) was added
dropwise and the mixture was stirred for another 15 minutes and
used in the following reaction.
[0112] Upon cooling by an ice-salt bath, stannous chloride
dihydrate (22.6 g, 100 mmol) was dissolved in 10 mL of concentrated
hydrochloric acid followed by addition of above mentioned spare
mixture. The reaction mixture was warmed up to room temperature and
reacted for 1.5 hours. Then the mixture was adjusted to pH 9 with
40% aqueous sodium hydroxide. The mixture was extracted with 400 mL
of ethyl acetate and the combined organic extracts were
concentrated under reduced pressure and dried to obtain the title
compound (5,6,7,8-tetrahydro-naphthalen-2-yl)-hydrazine 3h (2.19 g,
yield 53.7%) as a yellow oil.
[0113] MS m/z (ESI): 163 [M+1]
Step 2
5-Methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
[0114] (5,6,7,8-Tetrahydro-naphthalen-2-yl)-hydrazine 3h (2.0 g,
12.3 mmol) was dissolved in 50 mL of acetic acid followed by
addition of ethyl acetoacetate (1.57 mL, 12.3 mmol). The reaction
mixture was heated to 100.degree. C. overnight. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
to obtain the title compound
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
3i (1.58 g, yield 56.2%) as a colourless oil.
[0115] MS m/z (ESI): 457 [2M+1]
[0116] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.54.about.7.58
(m, 2H), 7.08.about.7.10 (d, J=8 Hz, 1H), 3.43 (s, 2H), .delta.
2.77.about.2.81 (m, 4H), 2.21 (s, 3H), 1.80.about.1.83 (m, 4H).
Step 3
2'-Hydroxy-3'-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)-1-
,5-dihydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic
acid
[0117] Upon cooling by an ice-water bath,
3'-amino-2'-hydroxy-biphenyl-3-carboxylic acid hydrobromide 1f (250
mg, 1.09 mmol) was dissolved in 10 mL of hydrochloric acid (1 N)
followed by addition of 10 mL of aqueous sodium nitrite (82 mg, 1.2
mmol) and
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
3i (249 mg, 1.09 mmol). Then the mixture was adjusted to pH 8 with
saturated aqueous sodium bicarbonate, followed by addition of 10 mL
of ethanol. The reaction was warmed up to room temperature
overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered,
dried and recrystallized from methanol to obtain the title compound
2'-hydroxy-3'-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic
acid 3 (59 mg, yield 11.6%) as a yellow solid.
[0118] MS m/z (ESI): 467 [M-1]
[0119] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.75 (m, 4H),
2.33 (s, 3H), 2.70 (m, 4H), 7.13 (m, 3H), 7.36 (m, 1H), 7.60 (m,
2H), 7.71 (m, 1H), 7.75 (m, 1H), 7.97 (d, J=7.6 Hz, 1H), 8.14 (s,
1H), 9.66 (s, 1H), 13.03 (br, 1H), 13.76 (s, 1H)
Example 4
5'-Fluoro-2'-hydroxy-3'-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthal-
en-2-yl)-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic
acid
##STR00080##
[0121] Upon cooling by an ice-water bath,
3'-amino-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic acid 2f (250
mg, 1.01 mmol) was dissolved in 10 mL of hydrochloric acid (1 N)
followed by addition of 10 mL of aqueous sodium nitrite (77 mg,
1.12 mmol) and
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
4i (230 mg, 1.01 mmol). The mixture was adjusted to pH 8 with
saturated aqueous sodium bicarbonate, followed by addition of 10 mL
of ethanol. The reaction was warmed up to room temperature
overnight and monitored by TLC until the disappearance of the
starting materials. The mixture was filtered, dried and
recrystallized from methanol to obtain the title compound
5'-fluoro-2'-hydroxy-3'-{N-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-
-naphthalen-2-yl)-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-car-
boxylic acid 4 (64 mg, yield 13.1%) as a red solid.
[0122] MS m/z (ESI): 485 [M-1]
[0123] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.74 (m, 4H),
2.33 (s, 1H), 2.73 (m, 4H), 7.02 (dd, J.sub.1=9.2 Hz, J.sub.2=2.0
Hz, 1H), 7.11 (d, d=8.0 Hz, 1H), 7.47 (m, 1H), 7.63 (m, 3H), 7.82
(d, J=7.6 Hz, 1H), 7.98 (d, J=7.6 Hz, 1H), 8.18 (s, 1H), 9.58 (s,
1H), 13.05 (s, 1H), 13.62 (s, 1H)
Example 5
3'-[N'-(1-Bicyclo[4.2.0]octa-1(6),2,4-trien-3-yl-3-methyl-5-oxo-5-dihydro--
pyrazol-4-ylidene)-hydrazino]-2'-hydroxy-biphenyl-3-carboxylic
acid
##STR00081##
[0124] Step 1
3''-Bromo-bicyclo[4.2.0]octa-1(6),2,4-triene
[0125] Bicyclo[4.2.0]octa-1(6),2,4-triene 5a (7.9 g, 76 mmol) was
dissolved in 80 mL of water at room temperature. Upon cooling by an
ice-water, 3.9 mL of bromine was added dropwise. Upon completion of
the addition, the ice-water bath was removed and the reaction
mixture was warmed up to room temperature and stirred overnight.
The reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was diluted with 50 mL of n-hexane
and sodium sulfite (3 g, 23.8 mmol) was added. Upon completion of
the addition, the mixture was stirred at room temperature for 30
minutes. Then the separated organic layer was dried over anhydrous
sodium sulfate, filtered to remove the drying agent and
concentrated under reduced pressure to obtain the title compound
3-bromo-bicyclo[4.2.0]octa-1(6),2,4-triene 5b (13.53 g) as a
colourless oil, which was directly used in the next step.
[0126] MS m/z (ESI): 181.8 [M-1]
Step 2
N-(bicyclo[4.2.0]octa-1(6),2,4-trienyl-3-yl)-N'-(tert-butoxycarbonyl-hydra-
zino)-tert-butyl-carbonate
[0127] 3-Bromo-bicyclo[4.2.0]octa-1(6),2,4-triene 5b (13.5 g, 73.8
mmol) was dissolved in 100 mL of dry tetrahydrofuran. The mixture
was cooled to -78.degree. C. in a dry ice-ethanol bath and then
n-butyllithium (66 mL, 165 mmol) was added. A solution of
di-tert-butyl azodicarboxylate (20.1 g, 87.4 mmol) in 80 mL of dry
tetrahydrofuran was added dropwise under stirring at the same
temperature. Upon completion of the addition, the dry ice-ethanol
bath was removed and the mixture was warmed up to room temperature
and stirred overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The reaction was quenched
with 100 mL of water and the layers were separated. The aqueous
layer was extracted with ethyl acetate (100 mL.times.2). The
combined organic extracts were washed with saturated brine (150
mL.times.1), dried over anhydrous sodium sulfate and filtered to
remove the drying agent. The filtrate was purified by silica gel
column chromatography to obtain the title compound
N-(bicyclo[4.2.0]octa-1(6),2,4-trienyl-3-yl)-N'-(tert-butoxycarbonyl-hydr-
azino)-tert-butyl-carbonate 5c (4.07 g, 16.5%) as a yellow oil.
Step 3
2-Bicyclo[4.2.0]octa-1(6),2,4-trienyl-3-yl-5-methyl-2,4-dihydro-pyrazol-3--
one
[0128]
N-(bicyclo[4.2.0]octa-1(6),2,4-trienyl-3-yl)-N'-(tert-butoxycarbony-
l-hydrazino)-tert-butyl-carbonate 5c (4.0 g, 12 mmol) was dissolved
in 30 mL of acetic acid followed by addition of 30 mL of
trifluoroacetic acid and the mixture was stirred at room
temperature for 30 minutes followed by addition of 3-oxo-butanoic
acid methyl ester (1.6 mL, 15 mmol). The reaction mixture was
stirred at 100.degree. C. for 1.5 hours upon warming by an oil
bath. The reaction was monitored by TLC until the disappearance of
the starting materials. The reaction solvent was evaporated under
reduced pressure to dryness. Then 100 mL of water, 60 mL of ethyl
acetate and sodium carbonate (3 g) were added in batch. Upon
completion of the addition, the layers were separated. The aqueous
layer was extracted with ethyl acetate (40 mL.times.2). The
combined organic extracts were washed with saturated brine (100
mL.times.1), dried over anhydrous sodium sulfate, filtered to
remove the drying agent and concentrated under reduced pressure.
The residue was purified by silica gel column chromatography to
obtain the title compound
2-bicyclo[4.2.0]octa-1(6),2,4-trienyl-3-yl-5-methyl-2,4-dihydro-pyrazol-3-
-one 5d (910 mg, 37.9%) as a yellow solid.
[0129] MS m/z (ESI): 201.2 [M+1]
Step 4
[0130]
3'-[N'-(1-Bicyclo[4.2.0]octa-1(6),2,4-trien-3-yl-3-methyl-5-oxo-1,5-
-dihydro-pyrazol-4-ylidene)-hydrazino]-2'-hydroxy-biphenyl-3-carboxylic
acid
[0131] 3'-Amino-2'-hydroxy-biphenyl-3-carboxylic acid hydrobromide
1f (258 mg, 0.83 mmol) was dissolved in 10 mL of hydrochloric acid
(1 N), followed by addition of 10 mL of aqueous sodium nitrite (63
mg, 0.92 mmol) and
2-bicyclo[4.2.0]octa-1(6),2,4-trienyl-3-yl-5-methyl-2,4-dihydro-
-pyrazol-3-one 5d (150 mg, 0.75 mmol) upon cooling by an ice-water
bath. The mixture was adjusted to pH 8 with saturated aqueous
sodium bicarbonate, followed by addition of 10 mL of ethanol. The
reaction mixture was warmed up to room temperature overnight and
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered and the filter cake was dissolved in 30 mL
of water and adjusted pH about 3.about.4 with concentrated
hydrochloric acid. The mixture was then filtered and the filter
cake was washed with dichloromethane (8 mL). The residue was dried
to obtain the title compound
3'-[N'-(1-bicyclo[4.2.0]octa-1(6),2,4-trien-3-yl-3-methyl-5-oxo-1,5-dihyd-
ro-pyrazol-4-ylidene)-hydrazino]-2'-hydroxy-biphenyl-3-carboxylic
acid 5 (198 mg, 60%) as a red solid.
[0132] MS m/z (ESI): 439.5 [M-1]
[0133] .sup.1: NMR (400 MHz, DMSO-d.sub.6): .delta. 2.33 (s, 3H),
3.16 (m, 4H), 7.14 (m, 3H), 7.64 (m, 2H), 7.79 (m, 2H), 7.80 (m,
1H), 7.98 (m, 1H), 8.18 (s, 1H), 9.61 (s, 1H), 12.93 (br, 1H),
13.75 (br, 1H)
Example 6
5-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-1,2,4-trimethyl-1H-pyrro-
le-3-carboxylic acid ethyl ester
##STR00082## ##STR00083##
[0134] Step 1
2-(2-Methoxy-3-nitro-phenyl)-4,4,5,5-tetramethyl-[1,3,2]dioxaborolane
[0135] 1-Bromo-2-methoxy-3-nitro-benzene 1c (67 g, 0.289 mol),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi-1,3,2-dioxaborolane (110 g,
0.433 mol), tetrakis(triphenylphosphine)palladium (11.80 g, 14.44
mmol) and potassium acetate (71 g, 0.724 mol) were dissolved in 600
mL of ethylene glycol dimethyl ether. The mixture was heated to
reflux for 17 hours. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was
concentrated under reduced pressure and the residue was purified by
silica gel column chromatography to obtain the title compound
2-(2-methoxy-3-nitro-phenyl)-4,4,5,5-tetramethyl-[1,3,2]dioxaborolane
6a (50.5 g, 61.9%) as a yellow crystal.
Step 2
3,5-Dimethyl-1H-pyrrole-2,4-dicarboxylic acid 4-ethyl ester
[0136] 3,5-Dimethyl-1H-pyrrole-2,4-dicarboxylic acid 2-tert-butyl
ester 4-ethyl ester 6b (5.34 g, 20 mmol) was dissolved in
trifluoroacetic acid (7.4 mL, 100 mmol) and the mixture was stirred
for 2 hours. The reaction was monitored by TLC until the
disappearance of the starting materials. Then 40 mL of water was
added. The mixture was filtered and the filter cake was washed with
dichloromethane and dried to obtain the title compound
3,5-dimethyl-1H-pyrrole-2,4-dicarboxylic acid 4-ethyl ester 6c
(3.65 g, yield 86.5%) as a pink solid.
[0137] MS m/z (ESI): 209.8 [M-1]
Step 3
5-Iodo-2,4-dimethyl-1-pyrrole-3-carboxylic acid ethyl ester
[0138] 3,5-dimethyl-1H-pyrrole-2,4-dicarboxylic acid 4-ethyl ester
6c (3.65 g, 17.3 mmol) was dissolved in the solvent mixture of 100
mL of dichloromethane and mL of water followed by addition of
potassium iodide (11.5 g, 69.2 mmol) and iodine (4.39 g, 17.3
mmol). Upon completion of the addition, the reaction mixture was
heated to reflux for 2 hours. The reaction was monitored by TLC
until the disappearance of the starting materials. The mixture was
cooled to room temperature and then 20 mL of water and 10 mL of
sodium thiosulfate (2 M) were added. The mixture was extracted with
dichloromethane (30 mL.times.3). The combined organic extracts were
washed with saturated brine, dried over anhydrous magnesium
sulfate, filtered to remove the drying agent and concentrated under
reduced pressure to obtain the title compound
5-iodo-2,4-dimethyl-1H-pyrrole-3-carboxylic acid ethyl ester 6d
(4.1 g, yield 80.8%) as an orange solid.
Step 4
5-Iodo-1,2,4-trimethyl-1H-pyrrole-3-carboxylic acid ethyl ester
[0139] 5-Iodo-2,4-dimethyl-1H-pyrrole-3-carboxylic acid ethyl ester
6d (4.1 g, 13.99 mmol) was dissolved in 80 mL of tetrahydrofuran
followed by addition of 4-methyl-benzenesulfonic acid methyl ester
(2.73 g, 14.69 mmol) and sodium tert-butoxide (2.02 g, 20.99 mmol).
Upon completion of the addition, the reaction mixture was stirred
for 0.5 hours and monitored by TLC until the disappearance of the
starting materials. The mixture was filtered and the filter cake
was washed with tetrahydrofuran. The filtrate was concentrated
under reduced pressure to obtain the title compound
5-iodo-1,2,4-trimethyl-1H-pyrrole-3-carboxylic acid ethyl ester 6e
(3.8 g, yield 88.6%) as a grey solid.
[0140] MS m/z (ESI): 308.1 [M+1]
[0141] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 4.287.about.4.340
(q, 2H), 3.561 (s, 3H), 2.618 (s, 3H), 2.888 (s, 3H), 1.369-1.405
(t, 3H)
Step 5
5-(3-Nitro-2-methoxy-phenyl)-1,2,4-trimethyl-1H-pyrrole-3-carboxylic
acid ethyl ester
[0142] 5-Iodo-1,2,4-trimethyl-1H-pyrrole-3-carboxylic acid ethyl
ester 6e (2.88 g, 9.38 mmol) was dissolved in 25 mL of 1,4-dioxane
followed by addition of
2-(2-methoxy-3-nitro-phenyl)-4,4,5,5-tetramethyl-[1,3,2]dioxaborolane
6a (3.6 g, 10.3 mmol), tetrakis(triphenylphosphine)palladium (270
mg, 0.234 mmol), sodium carbonate (1.99 g, 18.77 mmol) and 10 mL of
water. Upon completion of the addition, the reaction mixture was
heated to reflux for 3 hours. The reaction was monitored by TLC
until the disappearance of the starting materials. The mixture was
cooled to room temperature followed by addition of 30 mL of water
and extracted with ethyl acetate (30 mL.times.3). The combined
organic extracts were washed with saturated brine, dried over
anhydrous magnesium sulfate, filtered to remove the drying agent
and concentrated under reduced pressure to obtain the title
compound
5-(3-nitro-2-methoxy-phenyl)-1,2,4-trimethyl-1H-pyrrole-3-carbox-
ylic acid ethyl ester 6f (1.25 g, yield 40.4%) as a yellow oil.
[0143] MS m/z (ESI): 333.2 [M+1]
[0144] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.817.about.7.841
(m, 1H), 7.457.about.7.476 (m, 1H), 7.284.about.7.324 (m, 1H),
4.322.about.4.375 (q, 2H), 3.521 (s, 3H), 3.316 (s, 3H), 2.625 (s,
3H), 2.177 (s, 3H), 1.398.about.1.434 (t, 3H)
Step 6
5-(3-Amino-2-methoxy-phenyl)-1,2,4-trimethyl-1H-pyrrole-3-carboxylic
acid ethyl ester
[0145]
5-(3-Nitro-2-methoxy-phenyl)-1,2,4-trimethyl-1H-pyrrole-3-carboxyli-
c acid ethyl ester 6f (300 mg, 0.9 mmol) was dissolved in 5 mL of
ethyl acetate followed by addition of formamide (227 mg, 1.61 mmol)
and 60 mg of palladium on carbon. The reaction mixture was heated
to reflux for 1 hour. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was washed with ethyl acetate. The filtrate was
concentrated under reduced pressure to obtain the title compound
5-(3-amino-2-methoxy-phenyl)-1,2,4-trimethyl-1H-pyrrole-3-carboxylic
acid ethyl ester 6g (234 mg, yield 86%) as a white solid.
[0146] MS m/z (ESI): 303.4 [M+1]
[0147] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 6.968.about.7.006
(m, 1H), 6.817.about.6.840 (m, 1H), 6.595.about.6.618 (m, 1H),
4.311.about.4.364 (q, 2H), 3.381 (s, 3H), 3.315 (s, 3H), 2.615 (s,
3H), 2.181 (s, 3H), 1.391.about.1.426 (t, 3H)
Step 7
5-(3-Amino-2-hydroxy-phenyl)-1,2,4-trimethyl-1H-pyrrole-3-carboxylic
acid ethyl ester hydrobromide
[0148]
5-(3-Amino-2-methoxy-phenyl)-1,2,4-trimethyl-1H-pyrrole-3-carboxyli-
c acid ethyl ester 6g (210 mg, 0.69 mmol) was dissolved in 5 mL of
dichloromethane followed by addition of boron tribromide (1.39 mL,
2.78 mmol). The reaction mixture was reacted at room temperature
for 0.5 hours and monitored by TLC until the disappearance of the
starting materials. The reaction was quenched with methanol, and
the mixture was concentrated under reduced pressure followed by
addition of 50 mL of ethyl acetate and 15 mL of saturated aqueous
sodium bicarbonate. The mixture was mixed well and the layers were
separated. The organic layer was washed with saturated brine, dried
over anhydrous magnesium sulfate, filtered to remove the drying
agent and concentrated under reduced pressure to obtain the title
compound
5-(3-amino-2-hydroxy-phenyl)-1,2,4-trimethyl-1H-pyrrole-3-carboxylic
acid ethyl ester hydrobromide 6h (165 mg, yield 82.5%) as a white
solid.
[0149] MS m/z (ESI): 289.3 [M+1]
[0150] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.
7.313.about.7.396 (m, 1H), 7.098.about.7.117 (m, 1H),
6.993.about.7.032 (m, 1H), 4.173.about.4.227 (q, 2H), 3.221 (s,
3H), 1.979 (s, 3H), 1.242.about.1.295 (t, 3H)
Step 8
5-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-1,2,4-trimethyl-1H-pyrro-
le-3-carboxylic acid ethyl ester
[0151]
5-(3-Amino-2-hydroxy-phenyl)-1,2,4-trimethyl-1H-pyrrole-3-carboxyli-
c acid ethyl ester hydrobromide 6h (140 mg, 0.51 mmol) was
dissolved in 1.76 mL of hydrochloric acid (1 N) followed by
addition of 1 mL of aqueous sodium nitrite (39 mg, 0.56 mmol) and
the mixture was stirred for 20 minutes upon cooling by an ice-water
bath.
5-Methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
3i (105 mg, 0.46 mmol) was then added. The mixture was adjusted to
pH 8 with saturated aqueous sodium bicarbonate, followed by
addition of 1 mL of ethanol. The reaction was warmed up to room
temperature overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was dried and then dissolved in
dichloromethane. Then the mixture was washed with saturated brine
and the organic layer was concentrated under reduced pressure. The
residue was purified by silica gel column chromatography to obtain
the title compound
5-(2-hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)-
-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-1,2,4-trimethyl-1H-pyrr-
ole-3-carboxylic acid ethyl ester 6 (120 mg, 50.8%) as a red
solid.
[0152] MS m/z (ESI): 514.1 [M+1]
[0153] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.84 (m, 4H),
2.14 (s, 3H), 2.41 (s, 3H), 2.57 (s, 3H), 2.80 (m, 4H), 3.34 (s,
3H), 3.87 (s, 3H), 6.51 (m, 1H), 7.00 (d, J=7.6 Hz, 1H), 7.11 (m,
2H), 7.71 (m, 2H), 13.82 (br, 1H)
Example 7
3'-{N'-[1-(2,3-Dihydro-benzofuran-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-
-4-ylidene]-hydrazino}-2'-hydroxy-biphenyl-3-carboxylic acid
##STR00084##
[0154] Step 1
[0155] Bis(2,2,2-trichloroethyl)
1-(2,3-dihydrobenzofuran-5-yl)hydrazine-1,2-dicarboxylate
2,3-Dihydro-benzofuran 7a (0.6 mL, 5.32 mmol),
bis(2,2,2-trichloroethyl) hydrazine-1,2-dicarboxylate (1.96 g, 5.15
mmol) and zinc chloride (920 mg, 6.76 mmol) were dissolved in 40 mL
of dichloromethane. The reaction was reacted overnight at room
temperature and monitored by TLC until the disappearance of the
starting materials. Then the mixture was purified by silica gel
column chromatography to obtain the title compound
bis(2,2,2-trichloroethyl)
1-(2,3-dihydrobenzofuran-5-yl)hydrazine-1,2-dicarboxylate 7b (2.5
g, yield 96.6%) as a white solid.
Step 2
2-(2,3-Dihydro-benzofuran-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one
[0156] Bis(2,2,2-trichloroethyl)
1-(2,3-dihydrobenzofuran-5-yl)hydrazine-1,2-dicarboxylate 7b (2.9
g, 5.8 mmol) was dissolved in 50 mL of ethanol and 5 mL of methanol
followed by addition of zinc powder (10.8 g, 166 mmol) and aqueous
ammonium acetate (15 mL, 1 mol/L) and the mixture was reacted at
room temperature for 1 hour. Then ethyl acetoacetate (0.75 mL, 5.9
mmol) was added dropwise. The reaction mixture was heated to reflux
for 2 hours. The reaction was monitored by TLC until the
disappearance of the starting materials. The reaction was cooled to
room temperature and filtered. The filtrate was concentrated under
reduced pressure and the residue was purified by silica gel column
chromatography to obtain the title compound
2-(2,3-dihydro-benzofuran-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one
7c (706 mg, yield 56.5%) as a yellow solid,
[0157] MS m/z (ESI): 217 [M+1]
[0158] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.63 (s, 1H),
7.51 (d, J=1.2 Hz, 1H), 6.78 (d, J=8.8 Hz, 1H), 4.57 (t, J=8.8 Hz,
2H), 3.39 (s, 2H), 3.22 (t, J=8.4 Hz, 2H), 2.16 (s, 3H)
Step 3
3'-{N'-[1-(2,3-Dihydro-benzofuran-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-
-4-ylidene]-hydrazino}-2'-hydroxy-biphenyl-3-carboxylic acid
[0159] 3'-Amino-2'-hydroxy-biphenyl-3-carboxylic acid hydrobromide
1f (155 mg, 0.5 mmol) was dissolved in 1.7 mL of hydrochloric acid
(1 N) followed by dropwise addition of 0.6 mL of aqueous sodium
nitrite (36 mg, 0.53 mmol) and the mixture was stirred for 10
minutes upon cooling by an ice-water bath. Then
2-(2,3-dihydro-benzofuran-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one
7c (97 mg, 0.45 mmol) was added. The mixture was adjusted to pH 7
by batch addition of saturated aqueous sodium bicarbonate (630 mg,
7.5 mmol). The reaction mixture was warmed up to room temperature
overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was adjusted
to pH<5 with concentrated hydrochloric acid. The mixture was
filtered and dried to obtain the title compound
3'-{N'-[1-(2,3-dihydro-benzofuran-5-yl)-3-methyl-5-oxo-1,5-dihyd-
ro-pyrazol-4-ylidene]-hydrazino}-2'-hydroxy-biphenyl-3-carboxylic
acid 7 (131 mg, yield 63.9%) as a brown solid.
[0160] MS m/z (ESI): 455 [M-1]
[0161] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.33 (s, 3H),
3.24 (t, J=8.4 Hz, 2H), 4.56 (t, J=8.4 Hz, 2H), 6.82 (d, J=8.4 Hz,
1H), 7.16 (m, 2H), 7.61 (m, 2H), 7.72 (m, 2H), 7.80 (d, J=8.0 Hz,
1H), 7.96 (d, J=8.0 Hz, 1H), 8.13 (d, J=1.6 Hz, 1H), 9.64 (s, 1H),
13.01 (s, 1H), 13.75 (s, 1H)
Example 8
2-(3,3-Dimethyl-indan-5-yl)-4-{[2-hydroxy-3'-(1H-tetrazol-5-yl-biphenyl-3--
yl]hydrazono}-5-methyl-2,4-dihydro-pyrazol-3-one
##STR00085## ##STR00086##
[0162] Step 1
5-(3-Bromo-phenyl)-1H-tetrazole
[0163] 3-Bromo-benzonitrile 8a (18.2 g, 0.1 mol) and ammonium
chloride (5.9 g, 0.11 mol) were dissolved in 80 mL of
N,N'-dimethylformamide followed by addition of sodium azide (7.16
g, 0.11 mol) under argon atmosphere. The reaction mixture was
heated to 100.degree. C. and reacted overnight. Then the mixture
was cooled to 60.degree. C. and concentrated under reduced pressure
to remove N,N'-dimethylformamide. The residue was diluted with 100
mL of water and 4 mL of concentrated hydrochloric acid and stirred
for 1 hour. The mixture was filtered and dried to obtain the title
compound 5-(3-bromo-phenyl)-1H-tetrazole 8b (23 g) as a white
solid.
Step 2
5-(2'-Methoxy-biphenyl-3-yl)-1H-tetrazole
[0164] 5-(3-Bromo-phenyl)-1H-tetrazole 8b (20 g, 89 mmol) and
2-methoxybenzeneboronic acid (14.2 g, 93.3 mmol) were dissolved in
530 mL of 1,4-dioxane followed by addition of
tetrakis(tripaphenylphosphine)palladium (1.84 g) and sodium
carbonate (18.9 g, 178 mmol) under argon atmosphere. The reaction
mixture was heated to reflux overnight. The reaction was monitored
by TLC until the disappearance of the starting materials. The
mixture was concentrated under reduced pressure to remove
1,4-dioxane and then hydrochloric acid (200 mL, 6 mol/L) was added.
The mixture was cooled for 2 hours and the layers were separated.
The organic layer was collected and concentrated. The residue was
dissolved in 500 mL of ethyl acetate and washed with 250 mL of
water. The mixture was dried over anhydrous sodium sulfate and
concentrated under reduced pressure. The residue was dissolved in
25 mL of ethyl acetate and standing overnight. The mixture was
filtered to obtain the title compound
5-(2'-methoxy-biphenyl-3-yl)-1H-tetrazole 8c (15 g, yield 68.2%) as
a light yellow solid.
Step 3
3'-(1H-Tetrazol-5-yl)-biphenyl-2-ol
[0165] 5-(2'-Methoxy-biphenyl-3-yl)-1H-tetrazole 8c (15.5 g, 61.5
mol) was dissolved in 195 mL of acetic acid followed by addition of
195 mL of hydrobromic acid under argon atmosphere. The reaction
mixture was heated to reflux for 5 hours. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was cooled to room temperature overnight. The mixture
was filtered and the filter cake was dissolved in 500 mL of ethyl
acetate. The mixture was washed with water (500 mL.times.2), dried
over anhydrous sodium sulfate, filtered and concentrated under
reduced pressure. The residue was recrystallized from 100 mL of
ethyl acetate to obtain the title compound
3'-(1H-tetrazol-5-yl)-biphenyl-2-ol 8d (12 g, yield 82.8%) as a
white solid.
Step 4
3-Nitro-3'-(1H-tetrazol-5-yl)-biphenyl-2-ol
[0166] 3'-(1H-Tetrazol-5-yl)-biphenyl-2-ol 8d (3.5 g, 14.7 mmol)
was dissolved in 145 mL of ethanol under argon atmosphere. Fuming
nitric acid (0.565 mL, 13.2 mmol) was added dropwise at 35.degree.
C. After the reaction mixture was reacted at room temperature for 1
hour, 150 mL of water was added. After standing overnight, the
mixture was filtered. The filter cake was washed with 100 mL of
water and dissolved in 500 mL of ethyl acetate and 250 mL of water.
The separated organic layer was washed with saturated brine and
dried over anhydrous magnesium sulfate. The mixture was purified by
silica gel column chromatography to obtain the title compound
3-nitro-3'-(1H-tetrazol-5-yl)-biphenyl-2-ol 8e (1 g, yield 27.0%)
as a yellow solid.
[0167] MS m/z (ESI): 282 [M-1]
Step 5
3-Amino-3'-(1H-tetrazol-5-yl)-biphenyl-2-ol hydrochloride
[0168] 3-Nitro-3'-(1H-tetrazol-5-yl)-biphenyl-2-ol 8e (2.5 g, 8.83
mmol) was dissolved in 118 mL of ethanol and 78.6 mL of water
followed by addition of aqueous sodium hydroxide (2.95 mL., 3
mol/L) and 313 mg of palladium on carbon. The mixture was
hydrogenated for 3 hours in a hydrogenator under 3 atm. of
hydrogen. The reaction was monitored by TLC until the disappearance
of the starting materials. The mixture was filtered and then
hydrochloric acid (60 mL, 3 mol/L) was added to the filtrate. The
filtrate was concentrated under reduced pressure. The residue was
diluted with a small amount of water and filtered. The filter cake
was washed with water and n-hexane and dried to obtain the title
compound 3-amino-3'-(1H-tetrazol-5-yl)-biphenyl-2-ol hydrochloride
8f (2.33 g) as a brown solid.
[0169] MS m/z (ESI): 252 [M+1]
Step 6
Di-tert-butyl
1-(3,3-dimethyl-2,3-dihydro-1H-inden-5-yl)hydrazine-1,2-dicarboxylate
[0170] 6-Bromo-1,1-dimethyl-indan (WO2005066115) 8g (4.32 g, 19.27
mmol) was dissolved in 40 mL of tetrahydrofuran and then
butyllithium (15.67 mL, 1.6 mol/L, 25.05 mmol) was added dropwise
at -78.degree. C. After the reaction mixture was reacted for 40
minutes, a solution of di-tert-butyl azodicarboxylate (5.32 g,
23.12 mmol) in 30 mL of tetrahydrofuran was then added. The
reaction mixture was reacted for another 3 hours at -78.degree. C.
The reaction was monitored by TLC until the disappearance of the
starting materials and quenched with 5 mL of methanol. The mixture
was warmed up to room temperature and filtered by silica gel. The
filtrate was concentrated under reduced pressure and the residue
was purified by silica gel column chromatography to obtain the
title compound di-tert-butyl
1-(3,3-dimethyl-2,3-dihydro-1H-inden-5-yl)hydrazine-1,2-dicarboxylate
8h (2.70 g, yield 37.2%) as a yellow solid.
Step 7
2-(3,3-Dimethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one
[0171] Di-tert-butyl
1-(3,3-dimethyl-2,3-dihydro-1H-inden-5-yl)hydrazine-1,2-dicarboxylate
8h (2.70 g, 7.18 mmol) was dissolved in 100 mL of acetic acid
followed by addition of 20 mL of trifluoroacetic acid. After the
mixture was reacted at room temperature for 2 hours, ethyl
acetoacetate (0.98 g, 7.54 mmol) was added. Then the mixture was
heated to 100.degree. C. and reacted for 2 hours. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was cooled to room temperature and concentrated under
reduced pressure to remove acetic acid. The reaction mixture was
neutralized by saturated aqueous sodium bicarbonate. Then the
mixture was extracted with ethyl acetate. The combined organic
extracts were washed with saturated brine, dried over anhydrous
sodium sulfate, filtered and concentrated under reduced pressure.
The resulting residue was purified by silica gel column
chromatography to obtain the title compound
2-(3,3-dimethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 8i
(1.0 g, yield 47.7%) as a light brown solid,
[0172] MS m/z (ESI): 243 [M+1]
Step 8
2-(3,3-Dimethyl-indan-5-yl)-4-{[2-hydroxy-3'-(1H-tetrazol-5-yl)-biphenyl-3-
-yl]-hydrazono}-5-methyl-2,4-dihydro-pyrazol-3-one
[0173] 3-Amino-3'-(1H-tetrazol-5-yl)-biphenyl-2-ol hydrochloride 8f
(290 mg, 1.0 mmol) was dissolved in hydrochloric acid (3.4 mL, 1
mol/L) followed by dropwise addition of 1.2 mL of aqueous sodium
nitrite (73 mg, 1.05 mmol) upon cooling by an ice-water bath. After
the mixture was reacted at room temperature for 10 minutes,
2-(3,3-dimethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 8i
(218 mg, 0.9 mmol), sodium bicarbonate (1.26 g, 15 mmol) and 4.4 mL
of ethanol were added successively. Then the reaction mixture was
reacted for 10 hours. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was washed with 20 mL of water and then
dissolved in 20 mL of water. Upon cooling by an ice-water bath, the
mixture was adjusted to pH<5 with concentrated hydrochloric
acid. The mixture was filtered and dried to obtain the title
compound
2-(3,3-dimethyl-indan-5-yl)-4-{[2-hydroxy-3'-(1H-tetrazol-5-yl)-biphenyl--
3-yl]-hydrazono}-5-methyl-2,4-dihydro-pyrazol-3-one 8 (336 mg,
yield 73.8%) as a yellow solid.
[0174] MS m/z (ESI): 505 [M-1]
[0175] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.24 (m, 6H),
1.92 (t, J=7.2 Hz, 2H), 2.36 (s, 3H), 2.87 (t, J=7.2 Hz, 2H), 7.21
(m, 3H), 7.73 (m, 5H), 8.08 (d, J=7.6 Hz, 1H), 8.25 (s, 1H), 9.77
(s, 1H), 13.80 (s, 1H)
Example 9
5-{2-Hydroxy-3-[N'-1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylide-
ne)-hydrazino]-phenyl}-furan-2-carboxylic acid
##STR00087## ##STR00088##
[0176] Step 1
5-(2-Methoxy-3-nitro-phenyl)furan-2-carboxylic acid
[0177]
2-(2-Methoxy-3-nitro-phenyl)-4,4,5,5-tetramethyl-[1,3,2]dioxaborola-
ne 6a (10 g, 35.85 mmol), 5-bromofuran-2-carboxylic acid (5.47 g,
28.66 mmol), tetrakis(triphenylphosphine)palladium (2.07 g, 1.79
mmol) and sodium carbonate (7.60 g, 71.66 mmol) were dissolved in
the solvent mixture of 200 mL of 1,4-dioxane and 30 mL of water.
The reaction mixture was heated to reflux for 2.5 hours. The
reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was filtered and the filtrate was
concentrated under reduced pressure. The residue was diluted with
150 mL of water and adjusted to pH 3 with 1 N hydrochloric acid.
Then the mixture was filtered and the filter cake was washed with
50 mL of the solvent mixture of n-hexane/ethyl acetate (V/V=1:1).
The residue was dried to obtain the title compound
5-(2-methoxy-3-nitro-phenyl)furan-2-carboxylic acid 9a (4.23 g,
yield 56.1%) as a grey solid.
[0178] MS m/z (ESI): 262 [M-1]
Step 2
5-(3-Amino-2-methoxy-phenyl)-furan-2-carboxylic acid
[0179] 5-(2-methoxy-3-nitro-phenyl)furan-2-carboxylic acid 9a (4.23
g, 16.09 mmol) was dissolved in 125 mL of ethyl acetate followed by
addition of 423 mg of palladium on carbon and ammonium formate
(4.054 g, 64.35 mmol). The reaction mixture was heated to reflux
for 3.5 hours. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was
concentrated under reduced pressure and the resulting residue was
purified by silica gel column chromatography to obtain the title
compound 5-(3-amino-2-methoxy-phenyl)-furan-2-carboxylic acid 9b
(2.79 g, yield 74.4%) as a light green solid.
[0180] MS m/z (ESI): 232 [M-1]
Step 3
5-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid
hydrobromide
[0181] 5-(3-Amino-2-methoxy-phenyl)-furan-2-carboxylic acid 9b
(2.79 g, 11.97 mmol) was dissolved in 25 mL of dichloromethane
followed by dropwise addition of boron tribromide (23.9 mL, 2.0
mol/L). The reaction mixture was reacted at room temperature for 1
hour. The reaction was monitored by TLC until the disappearance of
the starting materials. The mixture was concentrated under reduced
pressure after mL of methanol was added. The residue was diluted
with 100 mL of ethyl acetate and stirred for 1 hour. Then the
mixture was filtered and the filter cake was dried to obtain the
title compound 5-(3-amino-2-hydroxy-phenyl)-furan-2-carboxylic acid
hydrobromide 9c (1.24 g, yield 47.2%) as a yellow solid.
[0182] MS m/z (ESI): 218 [M-1]
Step 4
[0183] 5-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid
hydrobromide 9c (300 mg, 1.0 mmol) was dissolved in hydrochloric
acid (3.4 mL, 1 mol/L) followed by dropwise addition of 1.2 mL of
aqueous sodium nitrite (73 mg, 1.05 mmol) upon cooling by an
ice-water bath. After the mixture was reacted for 10 minutes,
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (193 mg, 0.9
mmol), sodium bicarbonate (1.26 g, 15 mmol) and 4.4 mL of ethanol
were added successively. The mixture was reacted at room
temperature for 24 hours. The reaction was monitored by TLC until
the disappearance of the starting materials. The mixture was
filtered and the filter cake was washed with 20 mL of water and
then dissolved in 20 mL of water. Upon cooling by an ice-water
bath, the mixture was adjusted to pH<5 with concentrated
hydrochloric acid, filtered and dried to obtain the title compound
5-{2-hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-yli-
dene)-hydrazino]-phenyl}-furan-2-carboxylic acid 9 (287 mg, yield
71.8%) as a yellow solid.
[0184] MS m/z (ESI): 443 [M-1]
[0185] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.03 (m, 2H),
2.32 (s, 3H), 2.89 (m, 4H), 7.15 (m, 1H), 7.22 (t, J=8.0 Hz, 1H),
7.29 (d, J=8.0 Hz, 1H), 7.36 (d, J=3.6 Hz, 1H), 7.57 (m, 1H), 7.70
(m, 2H), 7.78 (s, 1H), 9.97 (s, 1H), 13.73 (s, 1H)
Example 10
4-{[2-Hydroxy-3'-(1H-tetrazol-5-yl)-biphenyl-3-yl]-hydrazono}-5-methyl-2-(-
5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
##STR00089##
[0187] 3-Amino-3'-(1f'-tetrazol-5-yl)-biphenyl-2-ol hydrochloride
8f (340 mg, 1.34 mmol) was dissolved in 3 mL of 1 N hydrochloric
acid followed by dropwise addition of 3 mL of aqueous sodium
nitrite (98 mg, 1.41 mmol) upon cooling by an ice-water bath. After
the mixture was reacted for 10 minutes,
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyra-
zol-3-one 3i (276 mg, 1.21 mmol), sodium bicarbonate (1.69 g, 20
mmol) and 3 mL of ethanol were added successively. The reaction
mixture was reacted at room temperature for 18 hours. The reaction
was monitored by TLC until the disappearance of the starting
materials. The mixture was filtered and the filter cake was washed
with 20 mL of water and then dissolved in 20 mL of water. Upon
cooling by an ice-water bath, the mixture was adjusted to pH
3.about.4 with concentrated hydrochloric acid. Then the mixture was
filtered and dried to obtain the title compound
4-{[2-hydroxy-3'-(1H-tetrazol-5-yl)-biphenyl-3-yl]-hydrazono}-5-methyl-2--
(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one 10
(208 mg, 31.6%) as a yellow solid.
[0188] MS m/z (ESI): 491 [M-1]
[0189] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 8.19 (1H, s),
7.99 (1H, s), 7.69 (2H, t, J=8.8), 7.49 (21, d, J=7.6), 7.15 (3H,
m), 2.75 (4H, m), 2.39 (3H, s), 1.75 (4H, m)
Example 11
2'-Hydroxy-5'-methyl-3'-{N'-3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthale-
n-2-yl)-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic
acid
##STR00090## ##STR00091##
[0190] Step 1
2-Bromo-4-methyl-6-nitro-phenol
[0191] Sodium nitrate (28 g, 0.33 mmol) was dissolved in the
solvent mixture of 70 mL of concentrated sulfuric acid and 210 mL
of water at -5.degree. C., followed by slowly dropwise addition of
2-bromo-4-methyl-phenol 11a (30.8 g, 0.165 mol). The reaction
mixture was reacted for 2 hours upon cooling by an ice-water bath.
The mixture was warmed up to room temperature and reacted for
another 1 hour. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was extracted
with 200 mL of ethyl acetate. The combined organic extracts were
washed with water (100 mL.times.5), dried over anhydrous sodium
sulfate, filtered and concentrated under reduced pressure. The
residue was purified by silica gel column chromatography to obtain
the title compound 2-bromo-4-methyl-6-nitro-phenol 11b (22.24 g,
yield 58.1%) as a yellow solid.
[0192] .sup.1H NMR (400 MHz, DMSO-d.sub.6): 6, 2.29 (s, 3H), 7.81
(m, 2H), 10.76 (s, 1H)
Step 2
1-Bromo-2-methoxy-5-methyl-3-nitro-benzene
[0193] 2-Bromo-4-methyl-6-nitro-phenol 11b (22.24 g, 95.9 mmol) was
dissolved in 150 mL of acetone, followed by addition of potassium
carbonate (15.9 g, 115 mmol) and iodomethane (13.7 mL, 220.6 mmol).
The reaction mixture was heated to reflux overnight. The reaction
was monitored by TLC until the disappearance of the starting
materials. The mixture was filtered and the filtrate was
concentrated under reduced pressure. The residue was diluted with
100 mL of ethyl acetate and filtered. The mixture was concentrated
under reduced pressure to obtain the title compound
1-bromo-2-methoxy-5-methyl-3-nitro-benzene 11c (23.1 g, yield
97.9%) as an orange solid.
Step 3
2'-Methoxy-5'-methyl-3-nitro-biphenyl-3-carboxylic acid
[0194] 1-Bromo-2-methoxy-5-methyl-3-nitro-benzene 11c (15.0 g, 61
mmol) and 3-carboxyphenylboronic acid (11.6 g, 70.1 mmol) were
dissolved in 200 mL of 1,4-dioxane followed by addition of
tetrakis(triphenylphosphine)palladium (2.8 g, 2.44 mmol) and 61 mL
of aqueous sodium carbonate (12.9 g, 122 mmol). The reaction
mixture was heated to reflux overnight. The reaction was monitored
by TLC until the disappearance of the starting materials. The
mixture was concentrated under reduced pressure and the residue was
diluted with 500 mL of water. The mixture was washed with the
solvent mixture of 150 mL of n-hexane and 150 mL of ethyl acetate
followed by ethyl acetate (300 mL.times.2). The aqueous layer was
adjusted to pH 1.about.2 with concentrated hydrochloric acid. The
mixture was filtered and the filter cake was dried to obtain the
title compound 2'-methoxy-5'-methyl-3-nitro-biphenyl-3-carboxylic
acid 11d (15.4 g, 88.1%) as a yellow solid.
[0195] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.40 (s, 3H),
3.42 (s, 3H), 7.58 (s, 1H), 7.58.about.7.75 (min, 1H), 7.75 (d,
J=1.6 Hz, 1H), 7.82.about.7.84 (m, 1H), 8.02 (d, J=8 Hz, 1H), 8.11
(d, J=1.6 Hz, 1H), 13.12 (s, 1H)
Step 4
2'-Hydroxy-5'-methyl-3'-nitro-biphenyl-3-carboxylic acid
[0196] 2'-Methoxy-5'-methyl-3'-nitro-biphenyl-3-carboxylic acid 11d
(11.2 g, 39.0 mmol) was dissolved in hydrobromic acid (250 mL,
40%). The reaction mixture was heated to reflux overnight. The
reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was cooled to room temperature and
filtered. The filter cake was washed with water and n-hexane and
dried to obtain the title compound
2'-hydroxy-5'-methyl-3'-nitro-biphenyl-3-carboxylic acid 11e (9.15
g, yield 85.9%) as a yellow solid.
[0197] MS m/z (ESI): 272 [M-1]
[0198] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.36 (s, 3H),
7.60 (m, 2H), 7.78 (d, J=8 Hz, 1H), 7.88 (d, J=1.2 Hz, 1H), 7.97
(d, J=8 Hz, 1H), 8.11 (s, 1H), 10.44 (s, 1H), 13.06 (s, 1H)
Step 5
3'-Amino-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic acid
hydrochloride
[0199] 2'-Hydroxy-5'-methyl-3'-nitro-biphenyl-3-carboxylic acid 11e
(9.15 g, 33.5 mmol) was dissolved in 200 mL of ethyl acetate
followed by addition of 2 g of palladium on carbon and ammonium
formate (8.45 g, 134 mmol). The reaction mixture was heated to
reflux for 30 minutes. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filtrate was acidified by hydrochloric acid. The mixture
was filtered and dried to obtain the title compound
3'-amino-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic acid
hydrochloride 11f (6.65 g, yield 71.0%) as a white solid.
[0200] MS m/z (ESI): 242 [M-1]
[0201] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.29 (s, 3H),
7.09 (d, J=1.6 Hz, 1H), 7.14 (d, J=1.6 Hz, 1H), 7.59 (t, J=8 Hz,
1H), 7.74 (dd, J.sub.1=6.4 Hz, J.sub.2=1.6 Hz, 1H), 7.94 (dd,
J.sub.1=6.4 Hz, J.sub.2=1.6 Hz, 1H), 8.07 (s, 1H)
Step 6
2'-Hydroxy-5'-methyl-3'-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthal-
en-2-yl)-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic
acid
[0202] 3'-Amino-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic acid
hydrochloride 11f (272 mg, 0.97 mmol) was dissolved in hydrochloric
acid (3.3 mL, 1 mol/L) followed by dropwise addition of 1.3 mL of
aqueous sodium nitrite (74 mg, 1.07 mmol) upon cooling by an
ice-water bath. After the mixture was reacted for 20 minutes,
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
3i (200 mg, 0.88 mmol), sodium bicarbonate (1.22 g, 14.6 mmol) and
2.1 mL of ethanol were added successively. The reaction mixture was
reacted at room temperature for 4 hours. The reaction was monitored
by TLC until the disappearance of the starting materials. The
mixture was filtered and the filter cake was washed with 20 mL of
water and then dissolved in 20 mL of water. Upon cooling by an
ice-water bath, the mixture was adjusted to pH<5 with
concentrated hydrochloric acid, filtered and dried to obtain the
title compound
2'-hydroxy-5'-methyl-3'-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphtha-
len-2-yl)-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic
acid 11 (170 mg, yield 40.2%) as a red solid.
[0203] MS m/z (ESI): 481 [M-1]
[0204] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.75 (m, 4H),
2.30 (s, 3H), 2.34 (s, 3H), 2.74 (m, 4H), 7.00 (d, J=1.2 Hz, 1H),
7.11 (d, J=8.0 Hz, 1H), 7.53 (s, 1H), 7.62 (m, 3H), 7.80 (d, J=7.6
Hz, 1H), 7.95 (d, J=7.6 Hz, 1H), 8.14 (s, 1H), 9.39 (s, 1H), 13.03
(s, 1H), 13.77 (s, 1H)
Example 12
5-(3-{N'-[1-(3,3-Dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-
-ylidene]-hydrazino}-2-hydroxy-phenyl)-thiophene-2-carboxylic
acid
##STR00092## ##STR00093##
[0205] Step 1
5-(2-Methoxy-3-nitro-phenyl)thiophene-2-carboxylic acid
[0206]
2-(2-Methoxy-3-nitro-phenyl)-4,4,5,5-tetramethyl-[1,3,2]dioxaborola-
ne 6a (10 g, 35.85 mmol), 5-bromothiophene-2-carboxylic acid (6.68
g, 32.2 mmol), tetrakis(triphenylphosphine)palladium (2.07 g, 1.79
mmol) and sodium carbonate (7.59 g, 71.6 mmol) were dissolved in
the solvent mixture of 200 mL of 1,4-dioxane and 30 mL of water.
The reaction mixture was heated to reflux for 1 hour. The reaction
was monitored by TLC until the disappearance of the starting
materials. The mixture was filtered and the filtrate was
concentrated under reduced pressure. The residue was diluted with
150 mL of water and adjusted to pH 3 with 1 N hydrochloric acid.
The mixture was filtered and the filter cake was washed with 50 mL
of the solvent mixture of n-hexane/ethyl acetate (V:V=1/1) and
dried to obtain the title compound
5-(2-methoxy-3-nitro-phenyl)thiophene-2-carboxylic acid 12a (7.7 g,
yield 77%) as a light yellow solid.
[0207] MS m/z (ESI): 277.9 [M-1]
Step 2
5-(3-Amino-2-methoxy-phenyl)-thiophene-2-carboxylic acid
[0208] 5-(2-methoxy-3-nitro-phenyl)thiophene-2-carboxylic acid 12a
(7.7 g, 27.6 mmol) was dissolved in 300 mL of ethyl acetate
followed by addition of 500 mg of palladium on carbon and ammonium
formate (6.96 g, 110 mmol). The reaction mixture was heated to
reflux for 4 hours. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was
concentrated under reduced pressure and the resulting residue was
purified by silica gel column chromatography to obtain the title
compound 5-(3-amino-2-methoxy-phenyl)-thiophene-2-carboxylic acid
12b (6.2 g, yield 90.1%) as a grey solid.
[0209] MS m/z (ESI): 248 [M-1]
Step 3
5-(3-Amino-2-hydroxy-phenyl)-thiophene-2-carboxylic acid
hydrobromide
[0210] 5-(3-Amino-2-methoxy-phenyl)-thiophene-2-carboxylic acid 12b
(2.2 g, 8.83 mmol) was dissolved in 20 mL of dichloromethane
followed by dropwise addition of boron tribromide (35 mL, 35.32
mmol/L). The reaction mixture was reacted at room temperature for
1.5 hours. The reaction was monitored by TLC until the
disappearance of the starting materials. Then 5 mL of methanol was
added and the mixture was concentrated under reduced pressure. The
residue was diluted with 100 mL of ethyl acetate and stirred for
1.5 hours. The mixture was filtered and the filter cake was dried
to obtain the title compound
5-(3-amino-2-hydroxy-phenyl)-thiophene-2-carboxylic acid
hydrobromide 12c (1.2 g, yield 57.1%) as a grey solid.
[0211] MS m/z (ESI): 234 [M-1]
Step 4
5-(3-{N'-[1-(3,3-Dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-
-ylidene]-hydrazino}-2-hydroxy-phenyl)-thiophene-2-carboxylic
acid
[0212] 5-(3-Amino-2-hydroxy-phenyl)-thiophene-2-carboxylic acid 12c
(171 mg, 0.62 mmol) was dissolved in 3 mL of hydrochloric acid (1
N) upon cooling by an ice-water bath, followed by dropwise addition
of 1 mL of aqueous sodium nitrite (47 mg, 0.68 mmol). After the
mixture was stirred for 20 minutes,
2-(3,3-dimethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 8i
(150 mg, 0.62 mmol) was added. The mixture was adjusted to pH
8.about.9 by batch addition of aqueous sodium bicarbonate (781 mg,
9.3 mmol). The generated bubbles were quenched with 2 mL of ethanol
and the mixture was warmed up to room temperature and stirred
overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was dissolved in 20 mL of water. The mixture
was adjusted to pH 3.about.4 with concentrated hydrochloric acid,
filtered and dried. The crude product was purified by HPLC to
obtain the title compound
5-(3-{N'-[1-(3,3-dimethyl-indan-5-yl)-3-methyl-1-oxo-1,5-dihydro-pyrazol--
4-ylidene]-hydrazino}-2-hydroxy-phenyl)-thiophene-2-carboxylic acid
12 (48 mg, yield 15.9%) as an orange solid.
[0213] MS m/z (ESI): 487 [M-1]
[0214] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.24 (t, J=8.6,
6H), 1.93 (t, J=7.0, 2H), 2.87 (t, J=7.0, 2H), 7.16 (m, J=6.0, 1H),
7.27 (d, J=4.2, 2H), 7.57 (d, J=8.0, 1H), 7.64 (d, J=4.0, 1H), 7.70
(t, J=8.4, 2H), 7.75 (d, J=4.0, 1H)
Example 13
3'-{N'-[1-(2,3-Dihydro-benzofuran-5-yl)-3-methyl-oxo-1,5-dihydro-pyrazol-4-
-ylidene]-hydrazino}-2'-hydroxy-5-methyl-biphenyl-3-carboxylic
acid
##STR00094##
[0216] 3'-Amino-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic acid
hydrochloride 11f (287 mg, 1.03 mmol) was dissolved in 3.5 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 1.5 mL of aqueous sodium nitrite (78 mg,
1.13 mmol). After the mixture was stirred for 20 minutes,
2-(2,3-dihydro-benzofuran-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one
7c (200 mg, 0.93 mmol) was added. The mixture was adjusted to pH
8.about.9 by batch addition of aqueous sodium bicarbonate (1.298 g,
15.45 mmol). The generated bubbles were quenched with 2 mL of
ethanol. The mixture was warmed up to room temperature and reacted
overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and then the filter cake was dissolved in 20 mL of water. After
mixing well, the mixture was adjusted to pH 3.about.4 with
concentrated hydrochloric acid, filtered and dried. The residue was
washed with 10 mL of the solvent mixture of
dichloromethane/methanol (V:V=1:1), and then the crude product was
purified by HPLC to obtain the title compound
3'-{N''-[1-(2,3-dihydro-benzofuran-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyraz-
ol-4-ylidene]-hydrazino}-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic
acid 13 (100 mg, yield 23.0%) as a red solid.
[0217] MS m/z (ESI): 469 [M-1]
[0218] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.34 (s, 3H),
2.36 (s, 3H), 3.24 (t, J=8.8 Hz, 2H), 4.57 (t, J=8.8 Hz, 2H), 6.82
(d, J=8.8 Hz, 1H), 7.00 (s, 1H), 7.54 (s, 1H), 7.61 (m, 2H), 7.74
(s, 1H), 7.80 (d, J=7.6 Hz, 1H), 7.95 (d, J=7.6 Hz, 1H), 8.13 (s,
1H), 9.38 (s, 1H), 13.02 (s, 1H), 13.76 (s, 1H)
Example 14
3'-{N'-[1-(2,3-Dihydro-benzofuran-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-
-4-ylidene]-hydrazino}-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic
acid
##STR00095##
[0220] 3'-Amino-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic acid
hydrochloride 2f (219 mg, 0.722 mmol) was dissolved in 3 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 1 mL of aqueous sodium nitrite (59 mg, 0.85
mmol). After the mixture was stirred for 20 minutes,
2-(2,3-dihydro-benzofuran-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one
7c (150 mg, 0.69 mmol) was added. The mixture was adjusted to pH
8.about.9 by batch addition of aqueous sodium bicarbonate (1.007 g,
11.57 mmol). The generated bubbles were quenched with 2 mL of
ethanol. The reaction mixture was warmed up to room temperature and
reacted overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was dissolved in 15 mL of water. After mixing
well, the mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid, filtered and dried. The crude product was
purified by HPLC to obtain the title compound
3'-{N'-[1-(2,3-dihydro-benzofuran-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazo-
l-4-ylidene]-hydrazino}-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic
acid 14 (65 mg, yield 20.0%) as a red solid.
[0221] MS m/z (ESI): 473 [M-1]
[0222] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.33 (s, 3H),
3.23 (t, J=8.8 Hz, 2H), 4.56 (t, J=8.8 Hz, 2H), 6.83 (d, J=8.4 Hz,
1H), 7.03 (m, 1H), 7.48 (m, 1H), 7.60 (m, 2H), 7.65 (s, 1H), 7.83
(d, J=8.0 Hz, 1H), 7.98 (d, J=8.0 Hz, 1H), 8.17 (s, 1H), 9.57 (s,
1H), 13.07 (s, 1H), 13.62 (s, 1H)
Example 15
5-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-furan-2-carboxylic
acid
##STR00096##
[0224] 5-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid
hydrobromide 9c (292 mg, 0.975 mmol) was dissolved in 3.3 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 1.3 mL of aqueous sodium nitrite (74 mg,
1.07 mmol). After the mixture was stirred for 20 minutes,
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
3i (200 mg, 0.88 mmol) was added. The mixture was adjusted to pH
8.about.9 by batch addition of aqueous sodium bicarbonate (1.226 g,
14.6 mmol). The generated bubbles were quenched with 2 mL of
ethanol. The reaction mixture was warmed up to room temperature and
reacted overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was dissolved in 20 ml, of water. After mixing
well, the mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid, filtered and dried. The crude product was
purified by HPLC to obtain the title compound
5-(2-hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)-
-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-furan-2-carboxylic
acid 15 (160 mg, yield 39.8%) as a red solid.
[0225] MS m/z (ESI): 457 [M-1]
[0226] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.76 (m, 4H),
2.33 (s, 3H), 2.75 (m, 4H), 7.13 (m, 2H), 7.22 (t, J=8.0 Hz, 1H),
7.37 (d, J=3.2 Hz, 1H), 7.56 (d, J=7.6 Hz, 1H), 7.63 (m, 2H), 7.71
(d, J=8.4 Hz, 1H)
Example 16
3'-{N'-[1-(3,3-Dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-y-
lidene]-hydrazino}-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic
acid
##STR00097##
[0228] 3'-Amino-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic acid
hydrochloride 11f (257 mg, 0.92 mmol) was dissolved in 3.1 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 1.2 mL of aqueous sodium nitrite (70 mg,
1.01 mmol). After the mixture was stirred for 20 minutes,
2-(3,3-dimethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 8i
(200 mg, 0.83 mmol) was added. The mixture was adjusted to pH
8.about.9 by batch addition of aqueous sodium bicarbonate (1.157 g,
13.8 mmol). The generated bubbles were quenched with 2 mL of
ethanol. The reaction mixture was warmed up to room temperature and
reacted overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was dissolved in 30 mL of water. After mixing
well, the mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid, filtered and dried. The crude product was
purified by HPLC to obtain the title compound
3'-{N'-[1-(3,3-dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4--
ylidene]-hydrazino}-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic acid
16 (160 mg, yield 39.0%) as an orange solid.
[0229] MS m/z (ESI): 495 [M-1]
[0230] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.25 (s, 6H),
1.92 (t=7.2 Hz, 2H), 2.35 (s, 3H), 2.86 (t, J=7.6 Hz, 2H), 7.00 (s,
1H), 7.25 (d, J=8.8 Hz, 1H), 7.54 (s, 1H), 7.61 (t, J=8.0 Hz, 1H),
7.69 (m, 2H), 7.80 (d, J=7.6 Hz, 1H), 7.95 (d, J=7.6 Hz, 1H), 8.14
(s, 1H), 9.41 (s, 1H), 13.05 (br, 1H), 13.77 (s, 1H)
Example 17
[0231]
3'-{N'-[1-(3,3-Dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyra-
zol-4-ylidene]-hydrazino}-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic
acid
##STR00098##
[0232] 3'-Amino-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic acid
hydrochloride 2f (200 mg, 0.71 mmol) was dissolved in 2.4 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 1 mL of aqueous sodium nitrite (54 mg, 0.78
mmol). After the mixture was stirred for 20 minutes,
2-(3,3-dimethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 8i
(153 mg, 0.64 mmol) was added. The mixture was adjusted to pH
8.about.9 by batch addition of aqueous sodium bicarbonate (889 mg,
10.6 mmol). The generated bubbles were quenched with 2 mL of
ethanol. The reaction mixture was warmed up to room temperature and
reacted overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was dissolved in 20 mL of water. After mixing
well, the mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid, filtered and dried. The crude product was
purified by HPLC to obtain the title compound
3'-{N'-[1-(3,3-dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4--
ylidene]-hydrazino}-5'-fluoro-2-hydroxy-biphenyl-3-carboxylic acid
17 (1.20 mg, yield 38.0%) as a red solid.
[0233] MS m/z (ESI): 499 [M-1]
[0234] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.25 (s, 6H),
1.92 (t, J=7.2 Hz, 2H), 2.34 (s, 3H), 2.86 (t, J=7.2 Hz, 2H), 7.03
(m, 1H), 7.25 (d, J=8.4 Hz, 1H), 7.47 (m, 1H), 7.63 (m, 3H), 7.83
(d, J=7.6 Hz, 1H), 7.98 (d, J=8.0 Hz, 1H), 8.18 (s, 1H), 9.60 (s,
1H), 13.07 (br, 1H), 13.64 (s, 1H)
Example 18
5-{2-Hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylid-
ene)-hydrazino]-phenyl}-thiophene-2-carboxylic acid
##STR00099##
[0236] 5-(3-Amino-2-hydroxy-phenyl)-thiophene-2-carboxylic acid
hydrobromide 12c (380 mg, 1.2 mmol) was dissolved in 3.9 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 1.5 mL of aqueous sodium nitrite (90 mg,
1.32 mmol). After the mixture was stirred for 20 minutes,
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (210 mg, 1 mmol)
was added. The mixture was adjusted to pH 8.about.9 by batch
addition of aqueous sodium bicarbonate (1.51 g, 18 mmol). The
generated bubbles were quenched with 2 mL of ethanol and the
reaction mixture was warmed up to room temperature and reacted
overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was dissolved in 20 mL of 5% aqueous sodium
hydroxide. The layers were separated and then the aqueous layer was
extracted with dichloromethane (50 mL.times.3). The aqueous layer
was adjusted to pH 3.about.4 with concentrated hydrochloric acid,
filtered and dried. The crude product was purified by HPLC to
obtain the title compound
5-{2-hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyra-
zol-4-ylidene)-hydrazino]-phenyl}-thiophene-2-carboxylic acid 18
(15 mg, yield 3.3%) as an orange solid.
[0237] MS m/z (ESI): 459 [M-1]
[0238] .sup.1H NMR (400 MHz, DMSO-d): .delta. 1.36.about.1.78 (m,
2H), 2.33 (s, 3H), 2.86.about.2.94 (m, 4H), 7.17 (t, J=8 Hz, 1H),
7.30 (d, J=8 Hz, 1H), 7.56 (dd, J.sub.1=8 Hz, J.sub.2=0.8 Hz, 1H),
7.56 (d, J=4 Hz, 1H), 7.70 (m, 2H), 7.75 (d, J=4 Hz, 1H), 7.79 (s,
1H)
Example 19
2'-Hydroxy-3'-{N'-[3-methyl-5-oxo-1-(1,1,3,3-tetramethyl-indan-5-yl)-1,5-d-
ihydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic acid
##STR00100## ##STR00101##
[0239] 5-Bromo-3,3-dimethyl-indan-1-one
[0240] 6-Bromo-1,1-dimethyl-indan 8g (4 g, 17.8 mmol) was dissolved
in 40 mL of anhydrous dichloromethane, followed by addition of
chromium oxide (280 mg, 1.8 mmol) and slowly dropwise addition of
tert-butyl hydroperoxide (19 mL, 190 mmol). The reaction system was
sealed and stirred overnight at room temperature after the reaction
solution was crimson and the generated gas was released. The
reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was diluted with 50 mL of water and
extracted with dichloromethane (100 mL.times.3). The combined
organic extracts were dried over anhydrous sodium sulfate, filtered
and concentrated under reduced pressure to obtain the title
compound 5-bromo-3,3-dimethyl-indan-1-one 19a (3.5 g, 82.3%) as a
white solid.
[0241] MS m/z (ESI): 238 [M-1]
Step 2
5-Bromo-1,1,3,3-tetramethyl-indan
[0242] Upon cooling by a dry ice-acetonitrile bath, 40 mL of
dichloromethane was cooled to -40.degree. C. followed by addition
of titanium tetrachloride (2.7 mL, 24.6 mmol) through a syringe.
The mixture was stirred for 20 minutes at the same temperature and
a solution of dimethyl zinc (29.3 mL, 35.1 mmol) in toluene was
added at such a rate that the reaction temperature was kept below
-30.degree. C. Upon completion of the addition, the reaction
mixture was stirred at the same temperature for 30 minutes and then
a solution of 5-bromo-3,3-dimethyl-indan-1-one 19a (2.8 g, 11.7
mmol) in 10 mL of dichloromethane was added. The reaction mixture
was warmed up to room temperature and reacted overnight. The
reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was concentrated under reduced
pressure and then the resulting residue was purified by silica gel
column chromatography to obtain the title compound
5-bromo-1,1,3,3-tetramethyl-indan 19b (1.55 g, 52.3%) as a
colourless oil.
[0243] MS m/z (ESI): 252 [M-1]
Step 3
Di-tert-butyl
1-(1,1,3,3-tetramethyl-2,3-dihydro-1H-inden-5-yl)hydrazine-1,2-dicarboxyl-
ate
[0244] 5-Bromo-1,1,3,3-tetramethyl-indan 19b (1.4 g, 5.53 mmol) was
dissolved in 10 mL of tetrahydrofuran and cooled to -78.degree. C.
in a dry ice-acetone bath. t-Butyllithium (4.4 mL, 11.1 mmol) was
added dropwise at the same temperature and the mixture was stirred
for 40 minutes. A solution of di-tert-butyl azodicarboxylate (1.59
g, 6.92 mmol) in 10 mL of tetrahydrofuran was added dropwise by a
constant-pressure addition funnel. The reaction mixture was stirred
at -78.degree. C. for 3 hours. The reaction was monitored by TLC
until the disappearance of the starting materials. The reaction was
quenched with 5 mL of methanol and was warmed up to room
temperature. The mixture was extracted with ethyl acetate (20
mL.times.3). The combined organic extracts were dried over
anhydrous sodium sulfate, filtered and concentrated under reduced
pressure. The residue was purified by silica gel column
chromatography to obtain the title compound di-tert-butyl
1-(1,1,3,3-tetramethyl-2,3-dihydro-1H-inden-5-yl)hydrazine-1,2-dicarboxyl-
ate 19c (1.326 g, yield 59.3%) as a yellow oil.
[0245] MS m/z (ESI): 403 [M+1]
Step 4
5-Methyl-2-(1,1,3,3-tetramethyl-indan-5-yl)-2,4-dihydro-pyrazol-3-one
[0246] Di-tert-butyl
1-(1,1,3,3-tetramethyl-2,3-dihydro-1H-inden-5-yl)hydrazine-1,2-dicarboxyl-
ate 19c (2.70 g, 7.18 mmol) was dissolved in 10 mL of acetic acid
followed by addition of 13 mL of trifluoroacetic acid. After the
mixture was reacted at room temperature for 30 minutes, ethyl
acetoacetate (502 mg, 3.86 mmol) was added. The reaction mixture
was heated to 100.degree. C. and reacted for 2 hours. The reaction
was monitored by TLC until the disappearance of the starting
materials. The mixture was cooled to room temperature and
concentrated under reduced pressure to remove acetic acid. The
mixture was neutralized by saturated aqueous sodium bicarbonate.
The mixture was extracted with ethyl acetate (20 mL.times.3). The
combined organic extracts were washed with saturated brine, dried
over anhydrous sodium sulfate, filtered and concentrated under
reduced pressure. The resulting residue was purified by silica gel
column chromatography to obtain the title compound
5-methyl-2-(1,1,3,3-tetramethyl-indan-5-yl)-2,4-dihydro-pyrazol-3-one
19d (130 mg, yield 13.6%) as a light yellow oil.
[0247] MS m/z (ESI): 269 [M+1]
Step 5
[0248]
2'-Hydroxy-3'-{N'-[3-methyl-5-oxo-1-(1,1,3,3-tetramethyl-indan-5-yl-
)-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic
acid
[0249] 3'-Amino-2'-hydroxy-biphenyl-3-carboxylic acid hydrobromide
1f (172 mg, 0.56 mmol) was dissolved in 1.9 mL of 1 N hydrochloric
acid upon cooling by an ice-water bath, followed by dropwise
addition of 0.7 mL of aqueous sodium nitrite (42 mg, 0.61 mm ol)
and
5-methyl-2-(1,1,3,3-tetramethyl-indan-5-yl)-2,4-dihydro-pyrazol-3-one
19d (135 mg, 0.5 mmol). The mixture was adjusted to pH 8.about.9 by
batch addition of saturated sodium bicarbonate (700 mg, 8.33 mmol)
followed by addition of 2 mL of ethanol. The reaction mixture was
warmed up to room temperature overnight. The reaction was monitored
by TLC until the disappearance of the starting materials. The
mixture was filtered and the filter cake was dissolved in mL of
water. After mixing well, the mixture was adjusted to pH 3.about.4
with concentrated hydrochloric acid, filtered and dried. The crude
product was purified by HPLC to obtain the title compound
2'-hydroxy-3'-{N'-[3-methyl-5-oxo-1-(1,1,3,3-tetramethyl-indan-5-yl)-1,5--
dihydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic acid 19
(75 mg, yield 29.4%) as a red solid.
[0250] MS m/z (ESI): 509 [M-1]
[0251] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.30 (m, 12H),
1.92 (s, 2H), 2.35 (s, 3H), 7.14 (m, 2H), 7.23 (d, J=8.0 Hz, 1H),
7.62 (m, 2H), 7.72 (m, 2H), 7.80 (d, J=7.6 Hz, 1H), 7.96 (d, J=8.0
Hz, 1H), 8.14 (s, 1H), 9.68 (s, 1H), 13.10 (br, 1H), 13.78 (s,
1H)
Example 20
2'-Hydroxy-5'-methyl-3'-{N'-[3-methyl-5-oxo-1-(1,1,3,3-tetramethyl-indan-5-
-yl)
1,5-dihydro-1-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic
acid
##STR00102##
[0253] 3'-Amino-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic acid
hydrochloride 11f (155 mg, 0.56 mmol) was dissolved in 1.9 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 1.2 mL of aqueous sodium nitrite (42 mg,
0.61 mmol). After the mixture was stirred for 20 minutes,
5-methyl-2-(1,1,3,3-tetramethyl-indan-5-yl)-2,4-dihydro-pyrazol-3-one
19d (135 mg, 0.5 mmol) was added. The mixture was adjusted to pH
8.about.9 by batch addition of aqueous sodium bicarbonate (700 mg,
8.33 mmol). The generated bubbles were quenched with 2 mL of
ethanol. The reaction mixture was warmed up to room temperature and
reacted overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was dissolved in 30 mL of water. After mixing
well, the mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid, filtered and dried. The crude product was
purified by HPLC to obtain the title compound
2'-hydroxy-5'-methyl-3'-{N'-[3-methyl-5-oxo-1-(1,1,3,3-tetramethyl-indan--
5-yl)-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic
acid 20 (50 mg, yield 19.1%) as a red solid.
[0254] MS m/z (ESI): 523 [M-1]
[0255] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.30 (m, 12H),
1.92 (s, 2H), 2.35 (m, 6H), 7.00 (m, 1H), 7.23 (d, J=8.4 Hz, 1H),
7.59 (s, 1H), 7.65 (m, 2H), 7.74 (m, 1H), 7.80 (d, J=8.0 Hz, 1H),
7.95 (d, J=7.6 Hz, 1H), 8.14 (s, 1H), 9.41 (s, 1H), 13.05 (br, 1H),
13.78 (s, 1H)
Example 21
3'-[N'-(1-Bicyclo[4.2.0]octa-1(6),2,4-trien-3-yl-3-methyl-5-oxo-1,5-dihydr-
o-9-pyrazol-4-ylidene)-hydrazino]-2'-hydroxy-5'-methyl-biphenyl-3-carboxyl-
ic acid
##STR00103##
[0257] 3'-Amino-2-hydroxy-5-methyl-biphenyl-3-carboxylic acid
hydrochloride 11f (311 mg, 1.11 mmol) was dissolved in 3.7 mL of 1
N hydrochloric acid upon cooling by an ice-water bath, followed by
dropwise addition of 1.5 mL of aqueous sodium nitrite (84 mg, 1.22
mmol) and
2-bicyclo[4.2.0]octa-1(6),2,4-trienyl-3-yl-5-methyl-2,4-dihydro-pyrazol-3-
-one 5d (200 mg, 1 mmol). The mixture was adjusted to pH 8.about.9
by batch addition of saturated sodium bicarbonate (1.4 g, 16.7
mmol) followed by addition of 2 mL of ethanol. The reaction mixture
was warmed up to room temperature overnight. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered and the filter cake was dissolved in 20
ml, of water. After mixing well, the mixture was adjusted to pH
3.about.4 with concentrated hydrochloric acid, filtered and dried.
The crude product was purified by HPLC to obtain the title compound
3'-[N'-(1-bicyclo[4.2.0]octa-1-(6),2,4-trien-3-yl-3-methyl-5-oxo-1,5-dihy-
dro-pyrazol-4-ylidene)-hydrazino]-2'-hydroxy-5'
methyl-biphenyl-3-carboxylic acid 21 (330 mg, yield 72.7%) as an
orange solid.
[0258] MS m/z (ESI): 453 [M-1]
[0259] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.34 (m, 6H),
3.16 (m, 4H), 7.00 (s, 1H), 7.15 (d, J=8.0 Hz, 1H), 7.54 (s, 1H),
7.62 (m, 2H), 7.71 (d, J=8.0 Hz, 1H), 7.80 (d, J=7.6 Hz, 1H), 7.95
(d, J=7.6 Hz, 1H), 8.13 (s, 1H), 9.40 (s, 1H), 13.02 (s, 1H), 13.75
(s, 1H)
Example 22
5-{3-[N'-(1-Bicylco-[4.2.0]octa-1(6),2,4-trien-3-yl-3-methyl-5-oxo-1,5-dih-
ydro-pyrazol-4-ylidene)-hydrazino]-2-hydroxy-phenyl}-furan-2-carboxylic
acid
##STR00104##
[0261] 5-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid
hydrobromide 9c (333 mg, 1.11 mmol) was dissolved in 3.7 mL of 1 N
hydrochloric acid upon cooling by an ice-water bath, followed by
addition of 1.5 mL of aqueous sodium nitrite (84 mg, 1.22 mmol) and
2-bicyclo[4.2.0]octa-1(6),2,4-trienyl-3-yl-5-methyl-2,4-dihydro-pyrazol-3-
-one 5d (200 mg, 1 mmol). The mixture was adjusted to pH 8.about.9
by batch addition of saturated sodium bicarbonate (1.4 g, 16.7
mmol), followed by addition of 2 mL of ethanol. The reaction
mixture was warmed up to room temperature overnight. The reaction
was monitored by TLC until the disappearance of the starting
materials. The mixture was filtered and the filter cake was
dissolved in 20 mL of water. After mixing well, the mixture was
adjusted to pH 3.about.4 with concentrated hydrochloric acid,
filtered and dried. The crude product was purified by HPLC to
obtain the title compound
5-{3-[N'-(1-bicyclo[4.2.0]octa-1(6),2,4-trien-3-yl-3-methyl-5-oxo-1,5-dih-
ydro-pyrazol-4-ylidene)-hydrazino]-2-hydroxy-phenyl}-furan-2-carboxylic
acid 22 (275 mg, yield 63.9%) as a red solid.
[0262] MS m/z (ESI): 429 [M-1]
[0263] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.31 (s, 3H),
3.15 (m, 4H), 7.15 (m, 2H), 7.20 (t, J=8.0 Hz, 1H), 7.36 (d, J=3.6
Hz, 1H), 7.54 (m, 1H), 7.68 (m, 1H), 7.72 (m, 2H), 9.97 (s, 1H),
13.71 (s, 1H)
Example 23
2-Bicylco[4.2.0]octa-1(6),2,4-trien-3-yl-4-{[2-hydroxy-3'-(1H-tetrazol-5-y-
l)-biphenyl-3-yl]-hydrazono}-5-methyl-2,4-dihydro-pyrazol-3-one
##STR00105##
[0265] 3-Amino-3'-(1H-tetrazol-5-yl)-biphenyl-2-ol hydrochloride 8f
(321 mg, 1.11 mmol) was dissolved in 3.7 mL of 1 N hydrochloric
acid upon cooling by an ice-water bath, followed by dropwise
addition of 1.5 mL of aqueous sodium nitrite (85 mg, 1.22 mmol).
After the mixture was reacted for 20 minutes,
2-bicyclo[4.2.0]octa-1(6),2,4-trienyl-3-yl-5-methyl-2,4-dihydro-pyrazol-3-
-one 5d (200 mg, 1 mmol), sodium bicarbonate (1.69 g, 20 mmol) and
3 mL of ethanol were added successively. The reaction mixture was
warmed up to room temperature and stirred overnight. The reaction
was monitored by TLC until the disappearance of the starting
materials. The mixture was filtered and the filter cake was washed
with 20 mL of water, and then dissolved in 20 mL of water. Upon
cooling by an ice-water bath, the mixture was adjusted to pH
3.about.4 with concentrated hydrochloric acid. The mixture was
filtered and dried to obtain the title compound
2-bicyclo[4.2.0]octa-1(6),2,4-trien-3-yl-4-{[2-hydroxy-3'-(1H-tetrazol-5--
yl)-biphenyl-3-yl]-hydrazono}-5-methyl-2,4-dihydro-pyrazol-3-one 23
(150 mg, yield 32.3%) as a red solid.
[0266] MS m/z (ESI): 463 [M-1]
[0267] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.34 (s, 3H),
3.16 (m, 4H), 7.18 (m, 3H), 7.68 (s, 1H), 7.73 (m, 4H), 8.08 (d,
J=7.6 Hz, 1H), 8.25 (d, J=10 Hz, 1H), 9.71 (br, 1H), 13.77 (br,
1H)
Example 24
5-{2-Hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylid-
ene)-hydrazino]-5-methyl-phenyl}-thiophene-2-carboxylic acid
##STR00106##
[0268] Step 1
2-(2-Methoxy-5-methyl-3-nitro-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborol-
ane
[0269] 3-Bromo-2-ethoxy-5-methyl-nitrobenzene 11c (20 g, 81.3 mmol)
and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi-1,3,2-dioxaborolane
(30.9 g, 112 mmol) were dissolved in 400 mL of dimethyl ether
followed by addition of
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (3.3 g,
4.06 mmol) and potassium acetate (19.9 g, 203 mmol). Upon
completion of the addition, the reaction mixture was heated to
reflux for 3 hours. The mixture was filtered to remove
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) and the
filtrate was concentrated under reduced pressure. The residue was
purified by silica gel column chromatography to obtain the title
compound
2-(2-methoxy-5-methyl-3-nitro-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaboro-
lane 24a (13.1 g, 57.1%) as a yellow oil.
[0270] MS m/z
Step 2
5-(3-Nitro-2-methoxy-5-methyl-phenyl)thiophene-2-carboxylic
acid
[0271]
2-(2-Methoxy-5-methyl-3-nitro-phenyl)-4,4,5,5-tetramethyl-1,3,2-dio-
xaborolane 24a (4.0 g, 14.5 mmol), 5-bromothiophene-2-carboxylic
acid (1.0 g, 4.8 mmol), tetrakis(triphenylphosphine)palladium
(0.276 g, 0.24 mmol) and sodium carbonate (1.01 g, 9.6 mmol) were
dissolved in 30 mL of 1,4-dioxane and 10 mL of water. The reaction
mixture was heated to reflux for 1 hour. The reaction was monitored
by TLC until the disappearance of the starting materials. The
mixture was concentrated under reduced pressure and the residue was
dissolved in water. The mixture was filtered to remove
tetrakis(triphenylphosphine)palladium and the filtrate was
extracted with ethyl acetate. The aqueous layer was acidified to
form precipitates. The precipitates were dissolved in ethyl
acetate, dried over anhydrous magnesium sulfate, filtered to remove
the drying agent and concentrated under reduced pressure to obtain
the title compound
5-(3-nitro-2-methoxy-5-methyl-phenyl)thiophene-2-carboxylic acid
24b (1.03 g, 73.6%) as a yellow solid.
[0272] MS m/z (ESI): 291.7 [M-1]
[0273] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 2.41 (s, 3H),
3.73 (s, 3H), 7.73.about.7.79 (m, 3H), 8.00 (m, 1H), 13.20 (br,
1H)
Step 3
5-(3-Amino-2-methoxy-5-methyl-phenyl)-thiophene-2-carboxylic
acid
[0274] 5-(3-Nitro-2-methoxy-5-methyl-phenyl)thiophene-2-carboxylic
acid 24b (0.29 g, 1 mmol) was dissolved in 30 mL of ethyl acetate
followed by addition of 0.06 g of palladium on carbon and ammonium
formate (0.25 g, 4 mmol). Upon completion of the addition, the
reaction mixture was heated to reflux for 45 minutes. The reaction
was monitored by TLC until the disappearance of the starting
materials. The mixture was filtered to remove palladium on carbon.
The filtrate was concentrated under reduced pressure and dried to
obtain the title compound
5-(3-amino-2-methoxy-5-methyl-phenyl)-thiophene-2-carboxylic acid
24c (0.26 g, yield 99%) as a green solid.
[0275] MS m/z (ESI): 261.7 [M-1]
[0276] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 2.18 (s, 3H),
3.58 (s, 3H), 6.54 (d, J=1.6 Hz, 1H), 6.78 (d, J=1.6 Hz, 1H), 7.53
(d, J=4 Hz, 1H), 7.68 (d, J=4 Hz, 1H)
Step 4
5-(3-Amino-2-hydroxy-5-methyl-phenyl)-thiophene-2-carboxylic acid
hydrobromide
[0277] 5-(3-Amino-2-methoxy-5-methyl-phenyl)-thiophene-2-carboxylic
acid 24c (0.26 g, 1 mmol) was dissolved in 20 mL of dichloromethane
followed by addition of boron tribromide (5 mL, 35.32 mmol/L). The
mixture was reacted at room temperature for 1 hour. The reaction
was monitored by TLC until the disappearance of the starting
materials. The mixture was concentrated under reduced pressure. The
residue was diluted with 30 mL of ethyl acetate and stirred for 0.5
hours. The mixture was filtered and the filter cake was washed with
ethyl acetate and dried to obtain the title compound
5-(3-amino-2-hydroxy-5-methyl-phenyl)-thiophene-2-carboxylic acid
hydrobromide 24d (0.15 g, yield 45.5%) as a grey solid.
[0278] MS m/z (ESI): 247.8 [M-1]
[0279] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 2.29 (s, 3H),
7.02 (d, J=1.6 Hz, 1H), 7.41 (s, 1H), 7.59 (d, J=4 Hz, 1H), 7.73
(d, J=4 Hz, 1H)
Step 5
5-{2-Hydroxy-3-[N-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylide-
ne)-hydrazino]-5-methyl-phenyl}-thiophene-2-carboxylic acid
[0280] 5-(3-Amino-2-hydroxy-5-methyl-phenyl)-thiophene-2-carboxylic
acid hydrobromide 24d (138 mg, 0.42 mmol) was dissolved in 1.5 mL
of 1 N hydrochloric acid upon cooling by an ice-water bath followed
by dropwise addition of 1.5 mL of aqueous sodium nitrite (85 mg,
1.22 mmol). After the mixture was reacted for 20 minutes,
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (81 mg, 0.38
mmol), sodium bicarbonate (527 mg, 6.27 mmol) and 2 mL of ethanol
were added successively. The reaction mixture was reacted overnight
at room temperature. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was washed with 20 mL of water, and then
dissolved in 10 mL of water. Upon cooling by an ice-water bath, the
mixture was adjusted to pH 3.about.4 with concentrated hydrochloric
acid, filtered and dried to obtain the title compound
5-{2-hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-yli-
dene)-hydrazino]-5-methyl-phenyl}-thiophene-2-carboxyli c acid 24
(30 mg, yield 16.7%) as a red solid.
[0281] MS m/z (ESI): 473 [M-1]
[0282] .sup.1H NMR (400 MHz, DMSO-d): .delta. 2.03 (m, 2H), 2.32
(s, 3H), 2.36 (s, 3H), 2.89 (m, 4H), 7.29 (d, J=8.0 Hz, 1H), 7.38
(s, 1H), 7.50 (s, 1H), 7.63 (d, J=4.0 Hz, 1H), 7.68 (d, J=8.0 Hz,
1H), 7.74 (m, 2H), 9.78 (s, 1H), 13.72 (br, 1H)
Example 25
5-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(1,1,3,3-tetramethyl-indan-5-yl)-1,5--
dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl-furan-2-carboxylic
acid
##STR00107##
[0284] 5-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid
hydrobromide 9e (124 mg, 0.41 mmol) was dissolved in 1.4 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 0.5 mL of aqueous sodium nitrite (32 mg,
0.45 mmol). After the mixture was stirred for 20 minutes,
5-methyl-2-(1,1,3,3-tetramethyl-indan-5-yl)-2,4-dihydro-pyrazol-3-one
19d (100 mg, 0.37 mmol) was added. The mixture was adjusted to pH 8
with saturated aqueous sodium bicarbonate. The generated bubbles
were quenched with 2 mL of ethanol. The reaction mixture was warmed
up to room temperature and reacted overnight. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered and the filter cake was dissolved in 10 mL
of water. The mixture was adjusted to pH 3.about.4 with
concentrated hydrochloric acid, filtered and dried. The residue was
purified by silica gel column chromatography to obtain the title
compound
5-(2-hydroxy-3-{N'-[3-methyl-5-oxo-1-(1,1,3,3-tetramethyl-indan-5-yl)-1,5-
-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-furan-2-carboxylic
acid 25 (22 mg, yield 11.9%) as a red solid.
[0285] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.30 (m, 12H),
1.92 (s, 2H), 2.34 (s, 3H), 7.15 (d, J=3.6 Hz, 1H), 7.22 (m, 2H),
7.36 (d, J=3.2 Hz, 1H), 7.56 (dd, J=8.0 Hz, J.sub.2=1.2 Hz, 1H),
7.66 (d, J=2.0 Hz, 1H), 7.75 (m, 2H)
Example 26
3'-[N'-(1-Bicyclo[4.2.0]octa-1(6),2,4-trien-3-yl-3-methyl-5-oxo-1,5-dihydr-
o-pyrazol-4-ylidene)-hydrazino]-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic
acid
##STR00108##
[0286] Step 1
3'-Amino-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic acid
hydrobromide
[0287] 20 mL of hydrobromic acid was added dropwise to
3'-amino-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic acid 2f (500
mg, 1.92 mmol) obtained from Step 5 of Example 2 to form white
precipitates. The reaction solution was heated to reflux overnight,
and then it became clear. The colour of the reaction solution
finally was brown. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was
concentrated under reduced pressure to obtain a brown solid which
was dissolved in 20 mL of ethyl acetate. The mixture was stirred
for 20 minutes and filtered to obtain the title compound
3'-amino-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic acid
hydrobromide 26a (344 mg, yield 54.8%) as a grey solid.
[0288] MS m/z (ESI): 248 [M+1]
Step 2
3'-[N'-(1-Bicyclo[4.2.0]octa-1(6),2,4-trien-3-yl-3-methyl-5-oxo-1,5-dihydr-
o-pyrazol-4-ylidene)-hydrazino]-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic
acid
[0289] 3'-Amino-5'-fluoro-2'-hydroxy-biphenyl-3-carboxylic acid
hydrobromide 26a (328 mg, 1.11 mmol) was dissolved in 3.7 mL of 1 N
hydrochloric acid upon cooling by an ice-water bath, followed by
addition of 1.5 mL of aqueous sodium nitrite (84 mg, 1.22 mmol) and
2-bicyclo[4.2.0]octa-1(6),2,4-trienyl-3-yl-5-methyl-2,4-dihydro-pyrazol-3-
-one 5d (200 mg, 1 mmol). The mixture was adjusted to pH 8.about.9
by batch addition of saturated sodium bicarbonate (1.4 g, 16.7
mmol), followed by addition of 2 mL of ethanol. The reaction
mixture was warmed up to room temperature overnight. The reaction
was monitored by TLC until the disappearance of the starting
materials. The mixture was filtered and then the filter cake was
dissolved in 20 mL of water. After mixing well, the mixture was
adjusted to pH 3.about.4 with concentrated hydrochloric acid,
filtered and dried. The crude product was purified by HPLC to
obtain the title compound
3'-[N'-(1-bicyclo[4.2.0]octa-1(6),2,4-trien-3-yl-3-methyl-5-oxo-1,5-dihyd-
ro-pyrazol-4-ylidene)-hydrazino]-5'-fluoro-2'-hydroxy-biphenyl-3-carboxyli-
c acid 26 (335 mg, yield 73.1%) as a red solid.
[0290] MS m/z (ESI): 457 [M-1]
[0291] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.32 (s, 3H),
3.22 (m, 4H), 7.02 (m, 1H), 7.14 (d, J=8.0 Hz, 1H), 7.46 (m, 1H),
7.61 (min, 2H), 7.69 (m, 1H), 7.82 (d, J=7, 6 Hz, 1H), 7.98 (d,
J=7.6 Hz, 1H), 8.17 (s, 1H), 9.58 (s, 1H), 13.07 (s, 1H), 13.60 (s,
1H)
Example 27
4-{[2-Hydroxy-3'-(1H-tetrazol-5-yl)biphenyl-3-yl]-hydrazono}-2-indan-5-yl--
5-methyl-2,4-dihydro-pyrazol-3-one
##STR00109##
[0293] 3-Amino-3'-(1H-tetrazol-5-yl)-biphenyl-2-ol hydrochloride 8f
(188 mg, 0.74 mmol) was dissolved in 4 mL of 2 N hydrochloric acid
upon cooling by an ice-water bath, followed by dropwise addition of
1 mL of aqueous sodium nitrite (57 mg, 0.82 mmol). After the
mixture was reacted for 30 minutes,
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (159 mg, 0.74
mmol) was added. The mixture was adjusted to pH 8 with saturated
sodium bicarbonate, followed by addition of 0.5 mL of ethanol. The
reaction mixture was reacted overnight at room temperature. The
reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was filtered and the filter cake
was dissolved in aqueous sodium hydroxide (3 N). The mixture was
washed three times with dichloromethane. The layers were separated
and then the aqueous layer was adjusted to pH 3 with 2 N
hydrochloric acid to form a copious amount of precipitates. The
mixture was filtered and the filter cake was purified by silica gel
column chromatography to obtain the title compound
4-{[2-hydroxy-3'-(H-tetrazol-5-yl)-biphenyl-3-yl]-hydrazono}-2-indan-5-yl-
-5-methyl-2,4-dihydro-pyrazol-3-one 27 (67 mg, yield 18.8%) as an
orange solid.
[0294] MS m/z (ESI): 477.2 [M-1]
[0295] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.03 (m, 2H),
2.33 (s, 3H), 2.89 (m, 4H), 7.17 (m, 2H), 7.28 (d, J=8.0 Hz, 1H),
7.71 (m, 5H), 8.08 (d, J=8.0 Hz, 1H), 8.25 (s, 1H), 9.73 (br, 1H),
13.77 (s, 1H)
Example 28
4-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-furan-2-carboxylic
acid
##STR00110## ##STR00111##
[0296] Step 1
4-Bromo-furan-2-carboxylic acid
[0297] A mixture of 4,5-dibromo-furan-2-carboxylic acid 28a (5.5 g,
20.3 mmol) and 18 mL of ammonium hydroxide was added to 63 mL of
water followed by addition of zinc powder (1.46 g, 22.33 mmol).
Upon completion of the addition, the reaction mixture was stirred
at room temperature for 6 hours. The reaction was monitored by TLC
until the disappearance of the starting materials. The mixture was
adjusted to pH 3 with hydrochloric acid (1 N) to form a copious
amount of precipitates. The mixture was filtered and the filter
cake was washed with n-hexane (15 mL.times.4) and dried to obtain
the title compound 4-bromo-furan-2-carboxylic acid 28b (3.2 g,
yield 83.1%) as a white solid.
[0298] MS m/z (ESI): 188.7 [M-1]
Step 2
4-(3-nitro-2-methoxy-phenyl)-furan-2-carboxylic acid
[0299]
2-(2-Methoxy-3-nitro-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolan-
e 6a (4 g, 14.34 mmol), 4-bromo-furan-2-carboxylic acid 28b (2.18
g, 11.47 mmol), tetrakis(triphenylphosphine)palladium (829 mg,
0.717 mmol) and potassium carbonate (3.96 g, 28.68 mmol) were
dissolved in the solvent mixture of 80 mL of 1,4-dioxane and 30 mL
of water. The reaction mixture was heated to reflux for 2.5 hours.
The reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was adjusted to pH 3 with 1 N
hydrochloric acid and then extracted with ethyl acetate (80
mL.times.3). The combined organic extracts were concentrated under
reduced pressure. The residue was purified by silica gel column
chromatography to obtain the title compound
4-(3-nitro-2-methoxy-phenyl)-furan-2-carboxylic acid 28c (3.42 g,
yield 90.7%) as a brown oil.
[0300] MS m/z (ESI): 261.8 [M-1]
Step 3
4-(3-Amino-2-methoxy-phenyl)-furan-2-carboxylic acid
[0301] 4-(3-nitro-2-methoxy-phenyl)-furan-2-carboxylic acid 28c
(500 mg, 1.9 mmol) was dissolved in 15 mL of ethyl acetate followed
by addition of 100 mg of palladium on carbon and ammonium formate
(429 mg, 7.6 mmol). The reaction mixture was heated to reflux for 3
hours. The reaction was monitored by TLC until the disappearance of
the starting materials. The mixture was filtered to remove
palladium on carbon and concentrated under reduced pressure to
obtain the title compound
4-(3-amino-2-methoxy-phenyl)-furan-2-carboxylic acid 28d (325 mg,
yield 73.4%) as a yellow oil.
[0302] MS m/z (ESI): 231.8 [M-1]
Step 4
4-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid
hydrobromide
[0303] 4-(3-Amino-2-methoxy-phenyl)-furan-2-carboxylic acid 28d
(325 mg, 1.4 mmol) was dissolved in 5 mL of dichloromethane
followed by dropwise addition of boron tribromide (2.8 mL, 5.6
mmol). The mixture was reacted at room temperature for 4.5 hours.
The reaction was monitored by TLC until the disappearance of the
starting materials. 5 mL of methanol was added and then the mixture
was concentrated under reduced pressure. The residue was diluted
with 10 mL of ethyl acetate and stirred for 0.5 hours. The mixture
was filtered and the filter cake was dried to obtain the title
compound 4-(3-amino-2-hydroxy-phenyl)-furan-2-carboxylic acid
hydrobromide 28e (174 mg, yield 57.1%) as a grey solid.
[0304] MS m/z (ESI): 217.7 [M-1]
Step 5
4-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-furan-2-carboxylic
acid
[0305] 4-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid 28e
(170 mg, 0.57 mmol) was dissolved in hydrochloric acid (1.9 mL, 1
mol/L) upon cooling by an ice-water bath, followed by dropwise
addition of 0.7 mL of aqueous sodium nitrite (43 mg, 0.63 mmol).
After the mixture was reacted for 20 minutes,
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
3i (116 mg, 0.51 mmol) was added. The mixture was adjusted to pH
8.about.9 with saturated aqueous sodium bicarbonate followed by
addition of 2 mL of ethanol. The mixture was reacted at room
temperature for 24 hours. The reaction was monitored by TLC until
the disappearance of the starting materials. The mixture was
filtered and then 15 mL of water was added to the filter cake. Upon
cooling by an ice-water bath, the mixture was adjusted to pH
2.about.3 with concentrated hydrochloric acid and filtered. The
filter cake was washed with ethyl acetate and dried to obtain the
title compound
4-(2-hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)-
-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-furan-2-carboxylic
acid 28 (13 mg, yield 5.5%) as a red solid.
[0306] MS m/z (ESI): 456.7 [M-1]
[0307] .sup.1H NMR (400 Mhz, DMSO-d.sub.5): .delta. 1.75 (m, 4H),
2.31 (s, 3H), 2.78 (m, 4H), 3.86 (s, 3H), 7.13 (m, 2H), 7.43 (d,
J=7.6 Hz, 1H), 7.62 (m, 2H), 7.78 (s, 1H), 8.43 (s, 1H), 9.68 (br,
1H), 13.73 (br, 1H)
Example 29
2'-Hydroxy-3'-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylide-
ne)-hydrazino]-5'-methyl-biphenyl-3-carboxylic acid
##STR00112##
[0309] 3-Amino-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic acid
hydrochloride 11f (346 mg, 1.07 mmol) was dissolved in hydrochloric
acid (5 mL, 2 mol/L) upon cooling by an ice-water, followed by
addition of 2 mL of aqueous sodium nitrite (81 mg, 1.17 mmol).
After the mixture was reacted for 30 minutes,
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (229 mg, 1.07
mmol) was added. The mixture was adjusted to pH 8 with saturated
aqueous sodium bicarbonate, followed by addition of 5 mL of
ethanol. The mixture was reacted overnight at room temperature. The
reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was filtered and the filter cake
was washed with ethanol. The filtrate was poured into a mixture of
ice and water. The resulting mixture was adjusted to pH 4 with
concentrated hydrochloric acid to form precipitates. The mixture
was filtered and the filter cake was washed three times with ethyl
acetate, and then dried to obtain the title compound
2'-hydroxy-3'-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylid-
ene)-hydrazino]-5'-methyl-biphenyl-3-carboxylic acid 29 (75 mg,
yield 15%) as a red solid.
[0310] MS m/z (ESI): 467.2 [M-1]
[0311] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.01 (m, 2H),
2.35 (m, 6H), 2.88 (m, 4H), 6.97 (s, 1H), 7.30 (s, 1H), 7.70 (m,
5H), 8.02 (s, 1H), 8.15 (s, 1H), 9.42 (br, 1H), 13.03 (br, 1H),
13.77 (s, 1H)
Example 30
3'-{N'-[1-(3,3)-Dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4--
ylidene]-drazino}-2'-hydroxy-biphenyl-3-carboxylic acid
##STR00113##
[0313] 3'-Amino-2'-hydroxy-biphenyl-3-carboxylic acid
hydrobrobromide 1f (310 mg, 1.0 mmol) was dissolved in hydrochloric
acid (3.4 mL, 1 mol/L) upon cooling by an ice-water bath, followed
by dropwise addition of 1.2 mL of aqueous sodium nitrite (73 mg,
1.05 mmol). After the mixture was reacted for 10 minutes,
2-(3,3-dimethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 8i
(218 mg, 0.9 mmol), sodium bicarbonate (1.26 g, 15 mmol) and 4.4 mL
of ethanol were added successively. The reaction mixture was
reacted at room temperature for 10 hours. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered and the filter cake was washed with 20 mL
of water and then dissolved in 20 mL of water. Upon cooling by an
ice-water bath, the mixture was adjusted to pH<5 with
concentrated hydrochloric acid, filtered and dried to obtain the
title compound
3'-{N'-[1-(3,3-dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4--
ylidene]-hydrazino}-2'-hydroxy-biphenyl-3-carboxylic acid 30 (500
mg, yield 94%) as a yellow solid.
[0314] MS m/z (ESI): 509.1 [M-1]
[0315] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.24 (m, 6H),
1.92 (t, J=7.2 Hz, 2H), 2.34 (s, 3H), 2.86 (t, J=7.2 Hz, 2H), 7.16
(m, 2H), 7.25 (d, J=8.8 Hz, 1H), 7.62 (t, J=7.6 Hz, 1H), 7.70 (m,
3H), 7.80 (d, J=7.6 Hz, 1H), 7.96 (d, J=7.2 Hz, 1H), 8.14 (s, 1H),
9.68 (s, 1H)
Example 31
4-{2-Hydroxy-3-[N'-Indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylidene-
)-hydrazino]-phenyl}-furan-2-carboxylic acid
##STR00114##
[0317] 4-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid
hydrobromide 28e (170 mg, 0.57 mmol) was dissolved in hydrochloric
acid (1.9 mL, 1 mol/L) upon cooling by an ice-water bath, followed
by dropwise addition of 0.7 mL of aqueous sodium nitrite (43 mg,
0.63 mmol). After the mixture was reacted for 20 minutes,
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (109 mg, 0.51
mmol) was added. The mixture was adjusted to pH 8.about.9 with
saturated aqueous sodium bicarbonate, followed by addition of 2 mL
of ethanol. The reaction mixture was reacted at room temperature
for 24 hours. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and 15 mL of water was added to the filter cake. Upon cooling by an
ice-water bath, the mixture was adjusted to pH 2.about.3 with
concentrated hydrochloric acid, filtered and the filter cake was
washed with ethyl acetate, and then dried to obtain the title
compound
4-{2-hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-yli-
dene)-hydrazino]-phenyl}-furan-2-carboxylic acid 31 (83 mg, yield
36.7%) as a black solid.
[0318] MS m/z (ESI): 442.8 [M-1]
[0319] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.08 (m, 2H),
2.32 (s, 3H), 2.89 (m, 4H), 7.13 (t, J=8.0 Hz, 1H), 7.28 (d, J=8.4
Hz, 1H), 7.41 (d, J=7.6 Hz, 1H), 7.65 (m, 3H), 7.75 (m, 1H), 8.37
(s, 1H), 9.68 (s, 1H), 13.22 (br, 1H), 13.74 (s, 1H)
Example 32
4-{[4'-(4,5-Dihydro-1H-imidazol-2-yl)-2-hydroxy-biphenyl-3-yl]-hydrazono}--
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one
##STR00115## ##STR00116##
[0320] Step 1
3'-Nitro-2'-methoxy-biphenyl-4-carbaldehyde
[0321] To a solution of 60 mL of 1,4-dioxane and 10 mL of water was
added 4-formylphenylboronic acid (3.0 g, 0.02 mol), followed by
1-bromo-2-methoxy-3-nitro-benzene 1c (4.64 g, 0.02 mol),
tetrakis(triphenylphosphine)palladium (1.15 g, 1 mmol) and sodium
carbonate (4.24 g, 0.04 mol). Upon completion of the addition, the
mixture was heated to reflux for 5 hours. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered and the filter cake was washed with ethyl
acetate. The filtrate was concentrated under reduced pressure and
the residue was purified by silica gel column chromatography to
obtain the title compound
3'-nitro-2'-methoxy-biphenyl-4-carbaldehyde 32a (4.1 g, 80.4%) as a
yellow solid.
Step 2
2-(2'-Methoxy-3'-nitro-biphenyl-4-yl)-4,5-dihydro-1H-imidazole
[0322] To a solution of 3'-nitro-2'-methoxy-biphenyl-4-carbaldehyde
32a (4.0 g, 15.5 mmol) in 40 mL of dichloromethane under stirring,
upon cooling by an ice-water bath, was added 1,2-diaminoethane (981
mg, 16.3 mmol). After the mixture was stirred for another 0.5
hours, 1-bromo-pyrrolidine-2,5-dione (2.91 g, 16.33 mmol) was
added. Upon completion of the addition, the ice-water bath was
removed. The reaction mixture was stirred overnight at room
temperature. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was
concentrated under reduced pressure and the residue was purified by
silica gel column chromatography to obtain the title compound
2-[(2'-methoxy-3'-nitro-biphenyl-4-yl]-4,5-dihydro-1H-imidazole 32b
(4.0 g, 86.9%) as yellow solid.
[0323] MS m/z (ESI): 298.1 [M+1]
[0324] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 8.11 (2H, d,
J=8.4 Hz), 7.89 (1H, dd, J.sub.1=8.0 Hz, J.sub.2=1.6 Hz), 7.87 (2H,
d, J=8.4 Hz), 7.79 (1H, m), 7.49 (1H, t, J=8.0 Hz), 4.04 (4H, m),
3.47 (3H, s)
Step 3
2-(2'-Methoxy-3'-amino-biphenyl-4-yl)-4,5-dihydro-1H-imidazol
[0325] To a solution of
2-[(2'-methoxy-3'-nitro-biphenyl-4-yl]-4,5-dihydro-1H-imidazole 32b
(1.5 g, 5.05 mmol) in 30 mL of methanol under stirring, was added
ammonium formate (1.28 g, 20.2 mmol) followed by palladium on
carbon (200 mg). Upon completion of the addition, the reaction
mixture was heated to reflux for 1 hour. The reaction was monitored
by TLC until the disappearance of the starting materials. The
mixture was cooled to room temperature and filtered. The filtrate
was concentrated under reduced pressure to obtain the title
compound
2-(2'-methoxy-3'-amino-biphenyl-4-yl)-4,5-dihydro-1H-imidazol 32c
(1.1 g, 81.5%) as a yellow solid.
[0326] MS m/z (ESI): 268.2 [M+1]
[0327] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 8.42 (2H, s),
7.96 (2H, d, J=8.0 Hz), 6.89 (1H, t, J=: 7.6 Hz), 6.75 (1H, m),
6.54 (1H, dd, J.sub.1=8.0 Hz, J.sub.2=1.6 Hz), 3.80 (4H, m), 3.47
(3H, s)
Step 4
2-(2'-Hydroxy-3'-amino-biphenyl-4-yl-4,5-dihydro-1H-imidazol
[0328]
2-(2'-Methoxy-3'-amino-biphenyl-4-yl)-4,5-dihydro-1H-imidazol 32c
(1.1 g, 4.1 mmol) was dissolved in 50 mL of dichloromethane at room
temperature, followed by addition of a solution of boron tribromide
in dichloromethane (1 N, 16.5 mL). The reaction mixture was stirred
at room temperature for 4 hours. The reaction was monitored by TLC
until the disappearance of the starting materials and then quenched
with methanol. The mixture was concentrated under reduced pressure.
The residue was dissolved in 10 mL of ethyl acetate and stirred for
30 minutes. The mixture was filtered and the filter cake was washed
with ethyl acetate (5 mL.times.2) and dried to obtained the title
compound
2-(2'-hydroxy-3'-amino-biphenyl-4-yl)-4,5-dihydro-1H-imidazol 32d
(900 mg, 89.6%) as a grey solid.
[0329] MS m/z (ESI): 254.3 [M+1]
[0330] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 7.23 (2H, m),
7.01 (2H, t, J=6.8 Hz), 6.65 (1H, m), 6.62 (1H, dd, J=8.0 Hz,
J.sub.2=1.6 Hz), 6.36 (1H, t, J=7.6 Hz), 4.03 (4H, m)
Step 5
4-{[4'-(4,5-Dihydro-1H-imidazol-2-yl)-2-hydroxy-biphenyl-3-yl]-hydrazono}--
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one
[0331] 2-(2'-Hydroxy-3'-amino-biphenyl-4-yl-4,5-dihydro-1H-imidazol
32d (334 mg, 1.11 mmol) was dissolved in hydrochloric acid (3.7 mm,
1 mol/L) upon cooling by an ice-water bath, followed by dropwise
addition of 1.5 mL of aqueous sodium nitrite (85 mg, 1.22 mmol).
After the mixture was reacted for 20 minutes,
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (214 mg, 1 mmol)
was added. The mixture was adjusted to pH 8 with saturated aqueous
sodium bicarbonate, followed by addition of 5 mL of ethanol. The
reaction mixture was reacted overnight at room temperature. The
reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was filtered and 30 mL of water was
added to the filter cake. After mixing well, the mixture was
adjusted to pH 4 with concentrated hydrochloric acid to form
precipitates. The mixture was filtered and the filter cake was
purified by silica gel column chromatography to obtain the title
compound
4-{[4'-(4,5-dihydro-1H-imidazol-2-yl)-2-hydroxy-biphenyl-3-yl]-hydrazono}-
-2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 32 (145 mg, yield
30.3%) as a red solid.
[0332] MS m/z (ESI): 476.9 [M-1]
[0333] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.02 (m, 2H),
2.34 (s, 3H), 2.87 (m, 4H), 4.00 (s, 4H), 6.88 (m, 1H), 7.18 (m,
2H), 7.49 (m, 1H), 7.90 (m, 5H), 8.18 (s, 1H)
Example 33
5-(2-Hydroxy-5-methyl-3{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthale-
n-2-yl)-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-thiophene-2-carb-
oxylic acid
##STR00117##
[0335] 5-(3-Amino-2-hydroxy-5-methyl-phenyl)-thiophene-2-carboxylic
acid hydrobromide 24d (366 mg, 1.11 mmol) was dissolved in 3.7 mL
of 1 N hydrochloric acid upon cooling by an ice-water bath,
followed by dropwise addition of 1.5 mL of aqueous sodium nitrite
(85 mg, 1.22 mmol). After the mixture was reacted for 20 minutes,
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
3i (228 mg, 1 mmol), sodium bicarbonate (1.4 g, 16.67 mmol) and 2
mL of ethanol were added successively. The reaction mixture was
reacted overnight at room temperature. The reaction was monitored
by TLC until the disappearance of the starting materials. The
mixture was filtered and the filter cake was added to 30 mL of
water. The mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid. The mixture was filtered and the filter cake was
dried and purified by silica gel column chromatography to obtain
the title compound
5-(2-hydroxy-5-methyl-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphtha-
len-2-yl)-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-thiophene-2-ca-
rboxylic acid 33 (88 mg, yield 18.09%) as a red solid.
[0336] MS m/z (ESI): 486.7 [M-1]
[0337] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.75 (min, 4H),
2.31 (s, 3H), 2.36 (s, 3H), 2.73 (m, 4H), 7.12 (d, J=8.4 Hz, 1H),
7.37 (s, 1H), 7.56 (s, 1H), 7.63 (m, 3H), 7.74 (d, J=3.6 Hz, 1H),
9.77 (s, 1H), 13.07 (br, 1H), 13.70 (s, 1H)
Example 34
5-(3-{N'-[1-(3,3-Dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-
-ylidene]-hydrazino}-2-hydroxy-5-methyl-phenyl)-thiophene-2-carboxylic
acid
##STR00118##
[0339] 5-(3-Amino-2-hydroxy-5-methyl-phenyl)-thiophene-2-carboxylic
acid hydrobromide 24d (366 mg, 1.11 mmol) was dissolved in 3.7 mL
of 1 N hydrochloric acid upon cooling by an ice-water bath,
followed by dropwise addition of 1.5 mL of aqueous sodium nitrite
(85 mg, 1.22 mmol). After the mixture was reacted for 20 minutes,
2-(3,3-dimethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 8i
(242 mg, 1 mmol) was added. The mixture was adjusted to pH 8 with
saturated aqueous sodium bicarbonate, followed by addition of 2 mL
of ethanol. Upon completion of the addition, the ice-water bath was
removed. The reaction mixture was reacted overnight at room
temperature. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was added to 30 mL of water. The mixture was
adjusted to pH 3.about.4 with concentrated hydrochloric acid, The
mixture was filtered and the filter cake was dried and purified by
silica gel column chromatography to obtain the title compound
5-(3-{N'-[1-(3,3-dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol--
4-ylidene]-hydrazino}-2-hydroxy-5-methyl-phenyl)-thiophene-2-carboxylic
acid 34 (308 mg, yield 61.4%) as a red solid.
[0340] MS m/z (ESI): 500.8 [M-1]
[0341] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.25 (s, 6H),
1.93 (min, 2H), 2.36 (s, 3H), 2.38 (s, 3H), 2.87 (t, J=7.2 Hz, 2H),
7.27 (d, J=8.8 Hz, 1H), 7.40 (d, J=1.6 Hz, 1H), 7.53 (d, J=1.2 Hz,
1H), 7.65 (m, 1H), 7.71 (m, 2H), 7.74 (d, J=4.0 Hz, 1H), 9.82 (br,
1H), 13.06 (s, 1H), 13.71 (br, 1H)
Example 35
5-{3-[N'-(1-Bicyclo[4.2.0]octa-1(6),2,4-trien-3-yl-3-methyl-5-oxo-1,5-dihy-
dro-pyrazol-4-ylidene)-hydrazino]-2-hydroxy-5-methyl-phenyl}-thiophene-2-c-
arboxylic acid
##STR00119##
[0343] 5-(3-Amino-2-hydroxy-5-methyl-phenyl)-thiophene-2-carboxylic
acid hydrobromide 24d (366 mg, 1.11 mmol) was dissolved in 3.7 mL
of 1 N hydrochloric acid upon cooling by an ice-water bath,
followed by dropwise addition of 1.5 mL of aqueous sodium nitrite
(85 mg, 1.22 mmol). After the mixture was reacted for 20 minutes,
2-bicyclo[4.2.0]octa-1(6),2,4-trienyl-3-yl-5-methyl-2,4-dihydro-pyrazol-3-
-one 5d (200 mg, 1 mmol) was added. The mixture was adjusted to pH
8 with saturated aqueous sodium bicarbonate, followed by addition
of 2 mL of ethanol. Then the ice-water bath was removed and the
reaction mixture was reacted at room temperature for 5 hours. The
reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was filtered and the filter cake
was added to 30 mL of water. The mixture was adjusted to pH
3.about.4 with concentrated hydrochloric acid and filtered. The
filter cake was washed with 6 mL of dichloromethane and dried to
obtain the title compound
5-{3-[N'-(1-bicyclo[4.2.0]octa-1(6),2,4-trien-3-yl-3-methyl-5-oxo-1,5-dih-
ydro-pyrazol-4-ylidene)-hydrazino]-2-hydroxy-5-methyl-phenyl}-thiophene-2--
carboxylic acid 35 (306 mg, yield 66.5%) as a red solid.
[0344] MS m/z (ESI): 459.2 [M-1]
[0345] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.33 (s, 3H),
2.36 (s, 3H), 3.16 (m, 4H), 7.16 (d, J=8.0 Hz, 1H), 7.38 (s, 1H),
7.49 (s, 1H), 7.62 (m, 2H), 7.73 (m, 2H)
Example 36
5-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-thiophene-2-carboxylic
acid
##STR00120##
[0347] 5-(3-Amino-2-hydroxy-phenyl)-thiophene-2-carboxylic acid
hydrobromide 12c (351 mg, 1.11 mmol) was dissolved in 3.7 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 1.5 mL of aqueous sodium nitrite (85 mg,
1.22 mmol). After the mixture was stirred for 20 minutes,
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
3i (228 mg, 1 mmol) was added. The mixture was adjusted to pH 8
with saturated aqueous sodium bicarbonate. The generated bubbles
were quenched with 2 mL of ethanol. The reaction was warmed up to
room temperature and reacted for 3 hours. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered and 30 mL of water was added to the filter
cake. The mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid. The mixture was filtered and the filter cake was
dried and purified by silica gel column chromatography to obtain
the title compound
5-(2-hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)-
-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-thiophene-2-carboxylic
acid 36 (30 mg, yield 6.3%) as a red solid.
[0348] MS m/z (ESI): 472.8 [M-1]
[0349] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.74 (m, 4H),
2.30 (s, 3H), 2.73 (m, 4H), 7.12 (d, J=8.0 Hz, 1H), 7.17 (m, 1H),
7.53 (d, J=7.6 Hz, 1H), 7.62 (m, 3H), 7.67 (d, J=8.0 Hz, 1H), 7.74
(d, J=4.0 Hz, 1H)
Example 37
5-{3-[N'-(1-Bicyclo[4.2.0]octa-1(6),2,4-trien-3-yl-3-methyl-5-oxo-1,5-dihy-
dro-pyrazol-4-ylidene)-hydrazino]-2-hydroxy-phenyl}-thiophene-2-carboxylic
acid
##STR00121##
[0351] 5-(3-Amino-2-hydroxy-phenyl)-thiophene-2-carboxylic acid 12c
(298 mg, 0.94 mmol) was dissolved in 3.1 mL of hydrochloric acid (1
N) upon cooling by an ice-water bath, followed by dropwise addition
of 1.3 ml of aqueous sodium nitrite (72 mg, 1.04 mmol). After the
mixture was reacted for 20 minutes,
2-bicyclo[4.2.0]octa-1(6),2,4-trienyl-3-yl-5-methyl-2,4-dihydro-pyrazol-3-
-one 5d (170 mg, 0.85 mmol) was added. The mixture was adjusted to
pH 8 with saturated aqueous sodium bicarbonate. The generated
bubbles were quenched with 2 mL of ethanol. The mixture was warmed
up to room temperature and reacted for 3 hours. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered and 30 mL of water was added to the filter
cake. The mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid and filtered. The filter cake was dried and
purified by silica gel column chromatography to obtain the title
compound
5-{3-[N'-(1-bicyclo[4.2.0]octa-1(6),2,4-trien-3-yl-3-methyl-5-oxo-1,5-dih-
ydro-pyrazol-4-ylidene)-hydrazino]-2-hydroxy-phenyl}-thiophene-2-carboxyli-
c acid 37 (57 mg, yield 15.01%) as a red solid,
[0352] MS m/z (ESI): 444.5 [M-1]
[0353] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.31 (s, 3H),
3.15 (m, 4H), 7.15 (m, 2H), 7.54 (d, J=7.6 Hz, 1H), 7.63 (m, 2H),
7.67 (d, J=7.6 Hz, 1H), 7.73 (m, 2H)
Example 38
2'-Hydroxy-3'-{N'-[3-methyl-1-(3-methyl-indan-5-yl)-5-oxo-1,5-dihydro-pyra-
zol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic acid
##STR00122## ##STR00123##
[0354] Step 1
6-Bromo-indan-1-one
[0355] To a 250 mL of flask was added 3-(4-bromo-phenyl)-propionic
acid 38a (20.6 g, 90 mmol, ABCR) and dried in vacuo for 20 minutes
followed by addition of 110 mL of anhydrous dichloromethane tinder
nitrogen atmosphere, and then thionyl chloride (20 mL, 276 mmol)
was added. The reaction mixture was heated to reflux overnight. The
mixture was concentrated under reduced pressure to remove most
solvent followed by addition of 100 mL, of dichloromethane, and
then aluminium trichloride (24.5 g, 25.8 mmol) was added. The
generated gas was released. The mixture was warmed up to reflux and
stirred overnight. The mixture was poured into 200 g of ice to form
a copious amount of precipitates and filtered through silica gel.
The filtrate was separated and the aqueous layer was extracted with
30 mL of dichloromethane. The combined organic extracts were dried
over anhydrous magnesium sulfate and concentrated under reduced
pressure to obtain the title compound 6-bromo-indan-1-one 38b
(18.34 g, yield 96.6%) as a yellow solid.
[0356] MS m/z (ESI): 210 [M-1]
Step 2
6-Bromo-1-methylene-indan
[0357] Methyltriphenyl phosphonium bromide (4.56 g, 12.76 mmol) was
dissolved in 25 mL of tetrahydrofuran followed by addition of
potassium tert-butoxide (1.5 g, 13.4 mmol). Upon completion of the
addition, the mixture was stirred at room temperature for 35
minutes and used in the following reaction.
[0358] 6-Bromo-indan-1-one 38b (898 mg, 4.25 mmol) was dissolved in
5 mL of tetrahydrofuran under stirring followed by addition of
above mentioned mixture. The reaction mixture was stirred at room
temperature for 1 hour prior to quenching by addition of 25 mL of
water. The mixture was extracted with dichloromethane (25
mL.times.4). The combined organic extracts were washed with
saturated brine (10 mL.times.2), dried over anhydrous sodium
sulfate, filtered and concentrated under reduced pressure. The
resulting residue was purified by silica gel column chromatography
to obtain the title compound 6-bromo-1-methylene-indan 38c (830 mg,
yield 93.4%) as a yellow oil.
[0359] MS m/z (ESI): 208 [M-1]
Step 3
6-Bromo-1-methyl-indan
[0360] 6-Bromo-1-methylene-indan 38c (4.91 g, 23.5 mmol) was
dissolve in ethyl acetate followed by addition of palladium on
carbon (0.98 g). The reaction mixture was reacted at room
temperature for 4 hours under hydrogen atmosphere. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered and the filtrate was concentrated under
reduced pressure. The residue was purified by silica gel column
chromatography to obtain the title compound 6-bromo-1-methyl-indan
38d (3.11 g, yield 62.7%) as a colourless oil.
[0361] MS m/z (ESI): 209.8 [M-1]
Step 4
Di-tert-butyl
1-(3-methyl-inden-5-yl)hydrazine-1,2-dicarboxylate
[0362] n-Butyllithium (8.6 mL, 13.76 mmol) was added to a
three-neck flask under argon atmosphere. Upon cooling by a dry
ice-acetone bath, 8 mL of tetrahydrofuran, 6-bromo-1-methyl-indan
38d (1.32 g, 6.26 mmol) were added successively under stirring. The
reaction mixture was reacted in the dry ice-acetone bath for 2
hours followed by dropwise addition of a solution of di-tert-butyl
azodicarboxylate (1.87 g, 8.14 mmol) in 10 mL of tetrahydrofuran.
The mixture was stirred for another 30 minutes and the ice-acetone
bath was removed. The mixture was warmed up to room temperature and
stirred for 20 hours. The reaction was quenched by addition of 20
mL of saturated ammonium chloride. The mixture was extracted with
ethyl acetate (25 mL.times.3). The combined organic extracts were
washed with saturated brine (10 mL.times.2), dried over anhydrous
sodium sulfate, filtered and concentrated under reduced pressure.
The residue was purified by silica gel column chromatography to
obtain the title compound di-tert-butyl
1-(3-methyl-inden-5-yl)hydrazine-1,2-dicarboxylate 38e (1.05 g,
yield 46.7%) as a yellow oil.
[0363] MS m/z (ESI): 362.6 [M+1]
Step 5
5-Methyl-2-(3-methyl-indan-5-yl)-2,4-dihydro-pyrazol-3-one
[0364] Di-tert-butyl
1-(3-methyl-inden-5-yl)hydrazine-1,2-dicarboxylate 38e (1.05 g, 2.9
mmol) was dissolved in 16 mL of a solvent mixture of ethanol and
water (V:V=5:3) under stirring, followed by successive addition of
ethyl acetoacetate (0.377 mL, 2.9 mmol) and 1.45 mL of 6 N
hydrochloric acid. The reaction mixture was heated to reflux and
reacted for 1.5 hours under nitrogen atmosphere. The mixture was
cooled to room temperature and the mixture was concentrated under
reduced pressure to remove ethanol. The aqueous layer was extracted
with dichloromethane (15 mL.times.3). The combined organic extracts
were dried over anhydrous sodium sulfate, filtered and concentrated
under reduced pressure. The residue was purified by silica gel
column chromatography to obtain the title compound
5-methyl-2-(3-methyl-indan-5-yl)-2,4-dihydro-pyrazol-3-one 38f (103
mg, yield 15.6%) as a yellow oil.
[0365] MS m/z (ESI): 229.3 [M+1]
Step 6
2'-Hydroxy-3'-{N'-[3-methyl-1-(3-methyl-indan-5-yl)-5-oxo-1,5-dihydro-pyra-
zol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic acid
[0366] 3'-Amino-2'-hydroxy-biphenyl-3-carboxylic acid hydrobromide
1f (344 mg, 1.11 mmol) was dissolved in 3.7 mL of hydrochloric acid
(1 N) upon cooling by an ice-water bath, followed by addition of
1.5 mL of aqueous sodium nitrite (85 mg, 1.22 mmol) and
5-methyl-2-(3-methyl-indan-5-yl)-2,4-dihydro-pyrazol-3-one 38f (228
mg, 1 mmol). The mixture was adjusted to pH 8 with saturated
aqueous sodium bicarbonate, followed by addition of 2 mL of
ethanol. The reaction mixture was warmed up to room temperature
overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and 30 mL of water was added to the filter cake. The mixture was
adjusted to pH 3.about.4 with concentrated hydrochloric acid and
filtered. The filter cake was dried and then 10 mL of a solvent
mixture of dichloromethane/methanol (V:V=50:1) was added. Upon
completion of the addition, the mixture was stirred for 1 hour.
Then the mixture was filtered and the filter cake was washed with
dichloromethane (2 mL.times.3) and dried to obtain the title
compound
2'-hydroxy-3'-{N'-[3-methyl-1-(3-methyl-indan-5-yl)-5-oxo-1,5-dihydro-pyr-
azol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic acid 38 (300 mg,
yield 64.1%) as a yellow solid.
[0367] MS m/z (ESI): 466.9 [M-1]
[0368] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.25 (m, 3H),
1.58 (m, 1H), 2.30 (m, 1H), 2.33 (s, 3H), 2.80 (m, 2H), 3.19 (m,
1H), 7.13 (m, 2H), 7.26 (d, J=8.0 Hz, 1H), 7.62 (t, J=7.6 Hz, 1H),
7.69 (m, 3H), 7.80 (d, J=7.6 Hz, 1H), 7.97 (d, J=7.6 Hz, 1H), 8.43
(s, 1H), 9.69 (br, 1H), 13.06 (s, 1H), 13.76 (s, 1H)
Example 39
2'-Hydroxy-5'-methyl-3'-{N'-[3-methyl-1-(3-methyl-indan-5-yl)-5-oxo-1,5-di-
hydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic acid
##STR00124##
[0370] 3'-Amino-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic acid
hydrochloride 11f (360 mg, 1.11 mmol) was dissolved in 3.7 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 1.5 mL of aqueous sodium nitrite (85 mg,
1.22 mmol). After the mixture was stirred for 20 minutes,
5-methyl-2-(3-methyl-indan-5-yl)-2,4-dihydro-pyrazol-3-one 38f (228
mg, 1.0 mmol) was added. The mixture was adjusted to pH 8 with
saturated aqueous sodium bicarbonate, followed by addition of 2 mL
ethanol. The reaction mixture was warmed up to room temperature and
reacted overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was dissolved in 30 mL of water. After mixing
well, the mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid. The mixture was filtered and the filter cake was
dried followed by addition of 10 mL dichloromethane. The mixture
was stirred for 1 hour and filtered. The filter cake was dried to
obtain the title compound
2'-hydroxy-5'-methyl-3'-{N'-[3-methyl-1-(3-methyl-indan-5-yl)-5-oxo-1,5-d-
ihydro-pyrazol-4-ylidene]-hydrazino}-biphenyl-3-carboxylic acid 39
(240 mg, yield 49.8%) as a red solid.
[0371] MS m/z (ESI): 480.9 [M-1]
[0372] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.29 (m, 3H),
1.56 (m, 1H), 2.28 (m, 1H), 2.34 (s, 3H), 2.36 (s, 3H), 2.80 (m,
2H), 3.18 (m, 1H), 7.00 (s, 1H), 7.27 (d, J=8.0 Hz, 1H), 7.54 (s,
1H), 7.61 (t, J=7.6 Hz, 1H), 7.70 (m, 2H), 7.80 (d, J=8.0 Hz, 1H),
7.95 (d, J=7.6 Hz, 1H), 8.14 (d, J=4.4 Hz, 1H), 9.40 (hr, 1H),
13.03 (s, 1H), 13.76 (s, 1H)
Example 40
5-(2-Hydroxy-3-{N'-[3-methyl-indan-5-yl)-5-oxo-1,5-dihydro-pyrazol-4-ylide-
ne]-hydrazino}-phenyl)-thiophene-2-carboxylic acid
##STR00125##
[0374] 5-(3-Amino-2-hydroxy-phenyl)-thiophene-2-carboxylic acid
hydrobromide 12c (351 mg, 1.11 mmol) was dissolved in 3.7 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 1.5 mL of aqueous sodium nitrite (85 mg,
1.22 mmol). After the mixture was stirred for 20 minutes,
5-methyl-2-(3-methyl-indan-5-yl)-2,4-dihydro-pyrazol-3-one 38f (228
mg, 1 mmol) was added. The mixture was adjusted to pH 8 with
saturated aqueous sodium bicarbonate. Then the generated bubbles
were quenched with 2 mL of ethanol. The reaction mixture was warmed
up to room temperature and reacted for 3 hours. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered and 30 mL of water was added to the filter
cake. The mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid. The mixture was filtered and the filter cake was
dried, and then purified by silica gel column chromatography to
obtain the title compound
5-(2-hydroxy-3-{N'-[3-methyl-1-(3-methyl-indan-5-yl)-5-oxo-1,5-dihydro-py-
razol-4-ylidene]-hydrazino}-phenyl)-thiophene-2-carboxylic acid 40
(90 mg, yield 19.0%) as a red solid.
[0375] MS m/z (ESI): 472.8 [M-1]
[0376] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.26 (m, 3H),
1.58 (m, 1H), 2.30 (m, 1H), 2.32 (s, 3H), 2.82 (m, 2H), 3.18 (m,
1H), 7.16 (t, J=8.0 Hz, 1H), 7.27 (d, J=8.0 Hz, 1H), 7.54 (d, J=7.6
Hz, 1H), 7.70 (m, 5H), 10.09 (br, 1H), 13.71 (br, 1H)
Example 41
3'-{N'-[1-(3-Ethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-yliden-
e]-hydrazino}-2'-hydroxy-biphenyl-3-carboxylic acid
##STR00126##
[0377] Step 1
6-Bromo-1-ethylidene-indan
[0378] (Ethyl)triphenylphosphonium bromide (14.5 g, 39.1 mmol) was
dissolved in 75 mL of tetrahydrofuran followed by addition of
potassium tert-butoxide (5.29 g, 47.3 mmol) at room temperature.
Upon completion of the addition, the reaction mixture was stirred
for 1 hour. A solution of 6-bromo-indan-1-one 38b (3.39 g, 18.6
mmol) in 25 mL of tetrahydrofuran was added to above mixture and
the mixture was stirred for another 1 hour. The reaction was
monitored by TLC until the disappearance of the starting materials.
The reaction was quenched with 150 mL of water and then the mixture
was extracted with dichloromethane (50 mL.times.4). The combined
organic extracts were washed with saturated brine (45 mL.times.2),
dried over anhydrous sodium sulfate, filtered to remove the drying
agent and concentrated under reduced pressure. The residue was
purified by silica gel column chromatography to obtain the title
compound 6-bromo-1-ethylidene-indan 41a (3.07 g, 74%) as a yellow
oil.
[0379] MS m/z (ESI): 221.8 [M-1]
Step 2
6-Bromo-1-ethyl-indan
[0380] 6-Bromo-1-ethylidene-indan 41a (3.07 g, 13.7 mmol) was
dissolved in 80 mL of ethyl acetate followed by addition of
palladium on carbon (0.61 g) at room temperature. The mixture was
hydrogenated for 5 hours in a hydrogenator under 3 atm. of
hydrogen. The reaction was monitored by TLC until the disappearance
of the starting materials. The mixture was filtered to remove
palladium on carbon and the filter cake was washed with ethyl
acetate (10 mL.times.3). The filtrate was concentrated under
reduced pressure and the residue was purified by silica gel column
chromatography to obtain the title compound 6-bromo-1-ethyl-indan
41b (2.75 g, 88.7%) as a light yellow oil.
[0381] MS m/z (ESI): 224 [M-1]
Step 3
Di-tert-butyl
1-(3-ethyl-2,3-dihydro-1H-inden-5-yl)hydrazine-1,2-dicarboxylate
[0382] Upon cooling by a dry ice-acetone bath, a solution of
6-bromo-1-ethyl-indan 41b (2.52 g, 11.2 mmol) in 15 mL of
tetrahydrofuran was added dropwise to a solution of t-butyllithium
in cyclohexane (18.1 mL, 1.3 N) under argon atmosphere. Upon
completion of the addition, the mixture was stirred for 2 hours in
the dry ice-acetone bath. A solution of di-tert-butyl
azodicarboxylate in 15 mL of tetrahydrofuran was added dropwise to
the above mixture at the same temperature. Upon completion of the
addition, the reaction mixture was stirred for another 1.5 hours in
the dry ice-acetone bath. Then the dry ice-ethanol bath was
removed. The mixture was warmed up to room temperature and stirred
for 18 hours. The reaction was monitored by TLC until the
disappearance of the starting materials and quenched by addition of
25 mL of saturated ammonium chloride. The mixture was extracted
with ethyl acetate (30 mL.times.3). The combined organic extracts
were dried over anhydrous sodium sulfate, filtered to remove the
drying agent and concentrated under reduced pressure. The residue
was purified by silica gel column chromatography to obtain the
title compound di-tert-butyl
1-(3-ethyl-2,3-dihydro-1H-inden-5-yl)hydrazine-1,2-dicarboxylate
41c (3.43 g, 81.5%) as a brown oil.
Step 4
2-(3-Ethylindan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one
[0383] Di-tert-butyl
1-(3-ethyl-2,3-dihydro-1H-inden-5-yl)hydrazine-1,2-dicarboxylate
41c (3.43 g, 9.1 mmol) was dissolved in 50 mL of a solvent mixture
of ethanol/water (V:V=3:2) followed by addition of 3-oxo-butanoic
acid ethyl ester (1.18 g, 9.1 mmol) and 4.55 mL of hydrochloric
acid (6 N). Upon completion of the addition, the reaction mixture
was heated to reflux and reacted for 1 hour. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was concentrated under reduced pressure to remove
ethanol and then extracted with dichloromethane (15 mL.times.3).
The combined organic extracts were dried over anhydrous sodium
sulfate, filtered to remove the drying agent and concentrated under
reduced pressure. The residue was purified by silica gel column
chromatography to obtain the title compound
2-(3-ethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 41d
(0.712 g, 39.8%) as a yellow oil.
[0384] MS m/z (ESI): 243.2 [M+1]
Step 5
3'-{N'-[1-(3-Ethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-yliden-
e]-hydrazino}-2'-hydroxy-biphenyl-3-carboxylic acid
[0385] 3'-Amino-2'-hydroxy-biphenyl-3-carboxylic acid hydrobromide
1f (285 mg, 0.92 mmol) was dissolved in 3.1 mL of hydrochloric acid
(1 N) upon cooling by an ice-water bath, followed by dropwise
addition of 1.2 mL of aqueous sodium nitrite (70 mg, 1.01 mmol).
After the mixture was stirred for 20 minutes,
2-(3-ethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 41d (200
mg, 0.83 mmol) was added. The mixture was adjusted to pH 8 with
saturated aqueous sodium bicarbonate followed by addition of 2 mL,
of ethanol. The reaction mixture was warmed up to room temperature
overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was added to 20 mL of water. The mixture was
adjusted to pH 3.about.4 with concentrated hydrochloric acid. The
mixture was filtered and the filter cake was dried to obtain the
title compound
3'-{N'-[1-(3-ethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylide-
ne]-hydrazino}-2'-hydroxy-biphenyl-3-carboxylic acid 41 (145 mg,
yield 36.4%) as a red solid.
[0386] MS m/z (ESI): 480.9 [M-1]
[0387] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 0.95 (m, 3H),
1.43 (m, 1H), 1.65 (m, 1H), 1.83 (m, 1H), 2.23 (m, 1H), 2.34 (s,
3H), 2.92 (m, 2H), 3.05 (m, 1H), 7.15 (s, 2H), 7.27 (d, J=8.0 Hz,
1H), 7.62 (t, J=8.0 Hz, 1H), 7.72 (m, 3H), 7.80 (d, J=7.6 Hz, 1H),
7.97 (d, J=8.0 Hz, 1H), 8.14 (s, 1H), 9.66 (s, 1H), 13.04 (s, 1H),
13.76 (s, 1H)
Example 42
3'-{N'-[1-(3-Ethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-yliden-
e]-hydrazino}-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic acid
##STR00127##
[0389] 3'-Amino-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic acid
hydrochloride 11f (298 mg, 0.92 mmol) was dissolved in 3.1 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 1.2 mL of aqueous sodium nitrite (70 mg,
1.01 mmol). After the mixture was stirred for 20 minutes,
2-(3-ethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 41d (200
mg, 0.83 mmol) was added. The mixture was adjusted to pH 8 with
saturated aqueous sodium bicarbonate followed by addition of 2 mL
of ethanol. The reaction mixture was warmed up to room temperature
overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and 20 mL of water was added to the filter cake. The mixture was
adjusted to pH 3.about.4 with concentrated hydrochloric acid. The
mixture was filtered and the filter cake was dried to obtain the
title compound
3'-{N-[1-(3-ethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-yliden-
e]-hydrazino}-2'-hydroxy-5'-methyl-biphenyl-3-carboxylic acid 42
(290 mg, yield 70.7%) as a red solid.
[0390] MS m/z (ESI): 494.9 [M-1]
[0391] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 0.95 (m, 3H),
1.43 (m, 1H), 1.65 (m, 1H), 1.83 (m, 1H), 2.23 (m, 1H), 2.34 (s,
3H), 2.92 (m, 2H), 3.05 (m, 1H), 6.99 (s, 1H), 7.26 (d, J=8.0 Hz,
1H), 7.53 (s, 1H), 7.60 (t, J=8.0 Hz, 1H), 7.68 (m, 1H), 7.75 (s,
1H), 7.80 (d, J=7.6 Hz, 1H), 7.96 (d, J=7.6 Hz, 1H), 8.14 (s, 1H),
9.39 (s, 1H), 13.03 (s, 1H), 13.76 (s, 1H)
Example 43
5-(3-{N'[1-(3,3-Dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4--
ylidene]-hydrazino}-2-hydroxy-phenyl)-furan-2-carboxylic acid
##STR00128##
[0393] 5-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid
hydrobromide 9c (333 mg, 1.1 mmol) was dissolved in hydrochloric
acid (3.7 mL, 1 mol/L) upon cooling by an ice-water bath, followed
by dropwise addition of 1.5 mL of aqueous sodium nitrite (85 mg,
1.22 mmol). After the mixture was reacted for 20 minutes,
2-(3,3-dimethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 8i
(242 mg, 1.0 mmol), sodium bicarbonate (1.4 g, 16.67 mmol) and 3 mL
of ethanol were added successively. The reaction mixture was
reacted overnight at room temperature. The reaction was monitored
by TLC until the disappearance of the starting materials. The
mixture was filtered and 20 mL of water was added to the filter
cake. The mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid. The mixture was filtered and the filter cake was
dried and purified by silica gel column chromatography to obtain
the title compound
5-(3-({N'-[1-(3,3-dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-
-4-ylidene]-hydrazino}-2-hydroxy-phenyl)-furan-2-carboxylic acid 43
(190 mg, yield 40.3%) as a red solid.
[0394] MS m/z (ESI): 470.9 [M-1]
[0395] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.26 (s, 6H),
1.92 (t, J=7.2 Hz, 2H), 2.33 (s, 3H), 2.86 (t, J=7.2 Hz, 2H), 7.15
(m, 1H), 7.20 (m, 2H), 7.37 (d, J=3.6 Hz, 1H), 7.55 (d, J=6.8 Hz,
1H), 7.71 (m, 3H), 9.99 (br, 1H), 13.15 (br, 1H), 13.74 (br,
1H)
Example 44
5-(2-Hydroxy-3-{N'-[3-methyl-1-(3-methyl-indan-5-yl)-5-oxo-1,5-dihydro-pyr-
azol-4-ylidene]-hydrazino}-phenyl)-furan-2-carboxylic acid
##STR00129##
[0397] 5-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid
hydrobromide 9c (333 mg, 1.1 mmol) was dissolved in hydrochloric
acid (3.7 mL, 1 mol/L) upon cooling by an ice-water bath, followed
by dropwise addition of 1.5 mL of aqueous sodium nitrite (85 mg,
1.22 mmol). After the mixture was reacted for 20 minutes,
5-methyl-2-(3-methyl-indan-5-yl)-2,4-dihydro-pyrazol-3-one 38c (228
mg, 1.0 mmol), sodium bicarbonate (1.4 g, 16.67 mmol) and 3 mL of
ethanol were added successively. The reaction mixture was reacted
overnight at room temperature. The reaction was monitored by TLC
until the disappearance of the starting materials. The mixture was
filtered and 20 mL of water was added to the filter cake. The
mixture was adjusted to pH 3.about.4 with concentrated hydrochloric
acid. The mixture was filtered and the filter cake was dried and
purified by silica gel column chromatography to obtain the title
compound
5-(2-hydroxy-3-{N'-[3-methyl-1-(3-methyl-indan-5-yl)-5-oxo-1,5-dihydro-py-
razol-4-ylidene]-hydrazino}-phenyl)-furan-2-carboxylic acid 44 (110
mg, yield 24.0%) as a red solid.
[0398] MS m/z (ESI): 457.0 [M-1]
[0399] .sup.1H NMR (400 MHz, DMSO-d): .delta. 1.25 (m, 3H), 1.59
(m, 1H), 2.29 (m, 1H), 2.33 (s, 3H), 2.84 (m, 2H), 3.17 (m, 1H),
7.14 (d, J=3.2 Hz, 1H), 7.22 (1, J=8.0 Hz, 1H), 7.28 (d, J=8.0 Hz,
1H), 7.36 (d, J=3.2 Hz, 1H), 7.55 (d, J=8.0 Hz, 1H), 7.71 (m, 3H)
9.96 (br, 1H) 13.75 (br, 1H)
Example 45
5-(3-{N'-[1-(2,2-Dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-
-ylidene]-hydrazino}-2-hydroxy-phenyl)-furan-2-carboxylic acid
##STR00130## ##STR00131##
[0400] Step 1
6-Bromo-2,2-dimethyl-indan-1-one
[0401] 6-Bromo-indan-1-one 38b (6.02 g, 28.5 mmol) and iodomethane
(4.4 mL, 70 mmol) were dissolved in 200 mL of dry tetrahydrofuran.
After the mixture was stirred at room temperature for 15 minutes,
sodium hydride (2.73 g, 68.2 mmol) was added. The reaction mixture
was stirred for another 2 hours. The reaction was monitored by TLC
until the disappearance of the starting materials. The reaction was
quenched with 150 mL of water and then extracted with ethyl acetate
(150 mL.times.2). The combined organic extracts were washed with
saturated brine, dried over anhydrous sodium sulfate, filtered to
remove the drying agent and concentrated under reduced pressure.
The residue was purified by silica gel column chromatography to
obtain the title compound 6-bromo-2,2-dimethyl-indan-1-one 45a
(5.36 g, yield 78.6%) as a brown solid.
[0402] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.918 (d, J=1.6
Hz, 1H), 7.725 (dd, J.sub.1=8 Hz, J.sub.2=2 Hz, 1H), 7.352 (d, J=8
Hz, 1H), 2.979 (s, 2H), 1.273 (s, 6H)
Step 2
5-Bromo-2,2-dimethyl-indan
[0403] 6-Bromo-2,2-dimethyl-indan-4-one 45a (7.23 g, 30.3 mmol) was
dissolved in 150 mL of trifluoroacetic acid followed by addition of
triethylsilane (12.1 mL, 75.6 mmol). The reaction mixture was
stirred overnight at room temperature. The reaction was monitored
by TLC until the disappearance of the starting materials and then
quenched by addition of water. The mixture was concentrated under
reduced pressure to remove trifluoroacetic acid. The mixture was
adjusted to be basic with saturated aqueous sodium bicarbonate and
then extracted with ethyl acetate (50 mL.times.3). The combined
organic extracts were washed with saturated brine, dried over
anhydrous sodium sulfate, filtered to remove the drying agent and
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography to obtain the title compound
5-bromo-2,2-dimethyl-indan 45b (14 g) as a colourless oil, which
was directly used in the next step.
[0404] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.335 (s, 1H),
7.281 (d. J=8 Hz, 1H), 7.063 (s, J=8 Hz, 1H), 2.749 (s, 2H), 2.702
(s, 2H), 1.188 (s, 6H)
Step 3
Di-tert-butyl 1-(2,2-dimethyl-5-yl)hydrazine-1,2-dicarboxylate
[0405] 5-Bromo-2,2-dimethyl-indan 45b (2.4 g, 10.7 mmol) was
dissolved in 20 mL of dry tetrahydrofuran. Upon cooling by a dry
ice-ethanol bath to -78.degree. C., n-butyllithium (12.1 mL, 30.2
mmol) was added dropwise. Upon completion of the addition, the
mixture was stirred for 2 hours. A solution of di-tert-butyl
azodicarboxylate (3.27 g, 14.2 mmol) in 20 mL of dry
tetrahydrofuran was added to the above mixture. The reaction
mixture was reacted at the same temperature for 3 hours. The
reaction was monitored by TLC until the disappearance of the
starting materials and quenched by addition of 50 mL of water. The
mixture was extracted with ethyl acetate (50 mL.times.2). The
combined organic extracts were washed with saturated brine, dried
over anhydrous sodium sulfate, filtered to remove the drying agent
and concentrated under reduced pressure. The residue was purified
by silica gel column chromatography to obtain the title compound
di-tert-butyl
1-(2,2-dimethyl-inden-5-yl)hydrazine-1,2-dicarboxylate 45c (2.68 g,
yield 66.8%) as a yellow oil.
Step 4
2-(2,2-Dimethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one
[0406] Di-tert-butyl
1-(2,2-dimethyl-1H-inden-5-yl)hydrazine-1,2-dicarboxylate
[0407] 45c (3.3 g, 8.78 mmol) was dissolved in 12 mL of acetic acid
followed by addition of 6 mL of trifluoroacetic acid and
3-oxo-butanoic acid methyl ester (1.5 mL, 13.8 mmol). Upon
completion of the addition, the reaction mixture was stirred at
90.degree. C. for 2 hours. The reaction was monitored by TLC until
the disappearance of the starting materials. The mixture was
concentrated under reduced pressure and the residue was diluted
with 30 mL of water and 30 mL of ethyl acetate. After mixing well,
the separated organic layer was washed with saturated brine, dried
over anhydrous sodium sulfate, filtered to remove the drying agent
and concentrated under reduced pressure. The residue was purified
by silica gel column chromatography to obtain the title compound
2-(2,2-dimethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 45d
(464 mg, yield 22.1%) as a yellow oil.
[0408] MS m/z (ESI): 243.3 [M1]
Step 5
5-(3-{N'-[1-(2,2-Dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-
-ylidene]-hydrazino}-2-hydroxy-phenyl)-furan-2-carboxylic acid
[0409] 5-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid
hydrobromide 9c (150 mg, 0.5 mmol) was dissolved in hydrochloric
acid (1.7 mL, 1 mol/L) upon cooling by an ice-water bath, followed
by dropwise addition of 0.6 mL of aqueous sodium nitrite (38 mg,
0.55 mmol). After the mixture was reacted for 20 minutes,
2-(2,2-dimethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 45d
(109 mg, 0.45 mmol), sodium bicarbonate (630 mg, 7.5 mmol) and 1 mL
of ethanol were added successively. The reaction mixture was
reacted overnight at room temperature. The reaction was monitored
by TLC until the disappearance of the starting materials. The
mixture was filtered and 15 mL, of water was added to the filter
cake. The mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid and filtered. The filter cake was washed with
ethyl acetate (1 mL.times.3) and dried to obtain the title compound
5-(3-{N'-[1-(2,2-dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol--
4-ylidene]-hydrazino}-2-hydroxy-phenyl)-furan-2-carboxylic acid 45
(67 mg, yield 31.6%) as a yellow solid.
[0410] MS m/z (ESI): 470.7 [M-1]
[0411] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.13 (s, 6H),
2.32 (s, 3H), 2.70 (m, 4H), 7.14 (d, J=3.6 Hz, 1H), 7.21 (m, 2H),
7.36 (d, J=3.6 Hz, 1H), 7.54 (m, 1H), 7.62 (m, 1H), 7.69 (m,
2H)
Example 46
5-{2-Hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylid-
ene)-hydrazino]-5-methyl-phenyl}-furan-2-carboxylic acid
##STR00132##
[0412] Step 1
5-(3-Nitro-2-methoxy-5-methyl-phenyl)-furan-2-carboxylic acid
[0413] To a solution of the solvent mixture of 30 mL of 1,4-dioxane
and 15 mL of water was added
2-(2-methoxy-5-methyl-3-nitro-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaboro-
lane 24a (3.1 g, 7.5 mmol) followed by 5-bromofuran-2-carboxylic
acid (1.3 g, 6.8 mmol), tetrakis (triphenylphosphine)palladium
(0.43 g, 0.4 mmol) and sodium carbonate (1.6 g, 15.1 mmol). The
reaction mixture was heated to reflux for 1 hour. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was cooled and filtered. The filtrate was washed with
ethyl acetate and concentrated under reduced pressure. The
resulting residue was diluted with 50 mL of water and adjusted to
pH 3 with concentrated hydrochloric acid and filtered. The filter
cake was washed with ethyl acetate and dried to obtain the title
compound 5-(3-nitro-2-methoxy-5-methyl-phenyl)-furan-2-carboxylic
acid 46a (522 mg, 29%) as a yellow solid.
[0414] MS m/z (ESI): 275.7 [M-1]
[0415] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 13.31 (1H, br),
7.90 (1H, s), 7.78 (1H, s), 7.39 (1H, d, J=3.2 Hz), 7.16 (1H, d,
J=3.6 Hz), 3.80 (3H, s), 2.42 (3H, s)
Step 2
5-(3-Amino-2-methoxy-5-methyl-phenyl)-furan-2-carboxylic acid
[0416] 5-(3-Nitro-2-methoxy-5-methyl-phenyl)-furan-2-carboxylic
acid 46a (410 mg, 1.48 mmol) was dissolved in 28 mL of ethyl
acetate followed by addition of 61 mg of palladium on carbon and
ammonium formate (658 mg, 10.44 mmol). Upon completion of the
addition, the reaction mixture was heated to reflux for 3 hours.
The reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was filtered to remove palladium on
carbon. The filtrate was concentrated under reduced pressure and
dried to obtain the title compound
5-(3-amino-2-methoxy-5-methyl-phenyl)-furan-2-carboxylic acid 46b
(356 mg, yield 97.3%) as a white solid.
[0417] MS m/z (ESI): 246.0 [M-1]
[0418] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 7.18 (1H, s),
6.98 (1H, m), 6.80 (1H, s), 6.56 (1H, s), 3.62 (3H, s), 2.20 (3H,
s)
Step 3
5-(3-Amino-2-hydroxy-5-methyl-phenyl)-furan-2-carboxylic acid
hydrobromide
[0419] 5-(3-Amino-2-methoxy-5-methyl-phenyl)-furan-2-carboxylic
acid 46b (248 mg, 1 mmol) was dissolved in 20 mL of dichloromethane
followed by dropwise addition of a solution of boron tribromide in
dichloromethane (3 mL, 1 mmol/L). The reaction mixture was reacted
at room temperature for 2 hours. The reaction was monitored by TLC
until the disappearance of the starting materials. The mixture was
concentrated under reduced pressure. The resulting residue was
washed with ethyl acetate (50 mL.times.3) and dried to obtain the
title compound
5-(3-amino-2-hydroxy-5-methyl-phenyl)-furan-2-carboxylic acid
hydrobromide 46c (172 mg, yield 54.7%) as a white solid.
[0420] MS m/z (ESI): 231.8 [M-1]
[0421] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 7.46 (1H, m),
7.34 (1H, m), 7.10 (2H, m), 2.31 (3H, s)
Step 4
5-{2-Hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylid-
ene)-hydrazino]-5-methyl-phenyl}-furan-2-carboxylic acid
[0422] 5-(3-Amino-2-hydroxy-5-methyl-phenyl)-furan-2-carboxylic
acid hydrobromide 46c (219 mg, 0.70 mmol) was dissolved in 2.3 mL
of 1 N hydrochloric acid upon cooling by an ice-water bath,
followed by dropwise addition of 1.0 mL of aqueous sodium nitrite
(53 mg, 0.77 mmol). After the mixture was reacted for 20 minutes,
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (134 mg, 0.63
mmol), sodium bicarbonate (878 mg, 10.45 mmol) and 2 mL of ethanol
were added successively. The reaction mixture was reacted overnight
at room temperature. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was added to 20 mL of water. The mixture was
adjusted to pH 3.about.4 with concentrated hydrochloric acid upon
cooling by an ice-water bath. The mixture was filtered and the
filter cake was washed with 6 mL of dichloromethane and dried to
obtain the title compound
5-{2-hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-yli-
dene)-hydrazino]-5-methyl-phenyl}-furan-2-carboxylic acid 46 (170
mg, yield 59.2%) as a red solid.
[0423] MS m/z (ESI): 456.8 [M-1]
[0424] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.05 (m, 2H),
2.32 (s, 3H), 2.37 (s, 3H), 2.88 (m, 4H), 7.13 (d, J=3.6 Hz, 1H),
7.28 (d, J=8.0 Hz, 1H), 7.35 (m, 2H), 7.51 (s, 1H), 7.68 (d, J=7.6
Hz, 1H), 7.78 (s, 1H)
Example 47
2-{2-Hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylid-
ene)-hydrazino]-phenyl}-5-methyl-thiazole-4-carboxylic acid
##STR00133## ##STR00134##
[0425] Step 1
2-(2-Methoxy-3-nitro-phenyl)-5-methyl-thiazole-4-carboxylic
acid
[0426]
2-(2-Methoxy-3-nitro-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolan-
e 6a (1.7 g, 6.08 mmol), 2-bromo-5-methyl-thiazole-4 carboxylic
acid (900 mg, 4.05 mmol), tetrakis (triphenylphosphine)palladium
(233 mg, 0.2 mmol) and sodium carbonate (1.29 g, 12.16 mmol) were
dissolved in 30 mL of 1,4-dioxane. The reaction mixture was heated
to reflux for 4 hours. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filtrate was concentrated under reduced pressure. The
residue was diluted with 20 mL of hydrochloric acid (1 N) and 30 mL
of ethyl acetate. The separated organic layer was washed with
saturated brine, dried over anhydrous sodium sulfate, filtered to
remove the drying agent and concentrated under reduced pressure.
The residue was recrystallized from a solvent mixture of ethyl
acetate and hexane to obtain the title compound
2-(2-methoxy-3-nitro-phenyl)-5-methyl-thiazole-4-carboxylic acid
47a (310 mg, yield 26%) as a yellow solid.
[0427] MS m/z (ESI): 292.6 [M-1]
[0428] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 13.45 (br, 1H),
8.58 (dd, J=8.0 Hz, 1H), 8.14 (dd, J=8.0 Hz, 1H), 7.52 (t, J=8.0
Hz, 1H), 3.93 (s, 3H), 2.71 (s, 3H)
Step 2
2-(2-Methoxy-3-amino-phenyl)-5-methyl-thiazole-4-carboxylic
acid
[0429] 2-(2-Methoxy-3-nitro-phenyl)-5-methyl-thiazole-4-carboxylic
acid 47a (300 mg, 1.02 mmol) was dissolved in 15 mL of methanol
followed by addition of 30 mg of palladium on carbon. The reaction
mixture was stirred for 24 hours under hydrogen atmosphere. The
reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was filtered to remove palladium on
carbon and the filtrate was concentrated under reduced pressure to
obtain the title compound
2-(2-methoxy-3-amino-phenyl)-5-methyl-thiazole-4-carboxylic acid
47b (250 mg, yield 92%) as a yellow solid.
[0430] MS m/z (ESI): 262.8 [M-1]
Step 3
2-(2-Hydroxy-3-amino-phenyl)-5-methyl-thiazole-4-carboxylic acid
hydrobromide
[0431] 2-(2-Methoxy-3-amino-phenyl)-5-methyl-thiazole-4-carboxylic
acid 47b (280 mg, 0.94 mmol) was dissolved in 5 mL of hydrogen
bromide. The reaction mixture was stirred overnight at 80.degree.
C. The reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was filtered and the filter cake
was washed with ethyl acetate and dried to obtain the title
compound
2-(2-hydroxy-3-amino-phenyl)-5-methyl-thiazole-4-carboxylic acid
hydrobromide 47c (200 mg, yield 64%) as a yellow solid.
[0432] MS m/z (ESI): 262.7 [M-1]
[0433] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 7.88 (d, J=8.0
Hz, 1H), 7.51 (d, J=8.0 Hz, 1H), 7.09 (t, J=8.0 Hz, 1H), 2.73 (s,
3H)
Step 4
2-{2-Hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylid-
ene)-hydrazino]-phenyl}-5-methyl-thiazole-4-carboxylic acid
[0434] 2-(2-Hydroxy-3-amino-phenyl)-5-methyl-thiazole-4-carboxylic
acid hydrobromide 47c (200 mg, 0.60 mmol) was dissolved in 2 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 0.82 mL of aqueous sodium nitrite (46 mg,
0.66 mmol). After the mixture was stirred for 20 minutes,
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (116 mg, 0.544
mmol) was added. The mixture was adjusted to pH 8.about.9 by batch
addition of saturated aqueous sodium bicarbonate (781 mg, 9.3
mmol). Then the generated bubbles were quenched with 2 mL of
ethanol. The reaction mixture was warmed up to room temperature and
reacted overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was dissolved in 20 mL of water. The mixture
was adjusted to pH 3.about.4 with concentrated hydrochloric acid.
The mixture was filtered and the filter cake was dried and purified
by HPLC to obtain the title compound
2-{2-hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-yli-
dene)-hydrazino]-phenyl}-5-methyl-thiazole-4-carboxylic acid 47
(195 mg, yield 75.9%) as a red solid.
[0435] MS m/z (ESI): 473.7 [M-1]
[0436] .sup.1H NMR (400 MHz, D.sub.2O): .delta. 1.97 (m, 2H), 2.31
(s, 3H), 2.53 (s, 3H), 2.80 (m, 4H), 6.52 (t, J=8.0 Hz, 1H), 7.23
(m, 4H), 7.84 (d, J=8.0 Hz, 1H)
Example 48
5-{2-Hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylid-
ene)-hydrazino]-phenyl}-1,2,4-trimethyl-1H-pyrrole-3-carboxylic
acid ethyl ester
##STR00135##
[0438] 5-(3-Amino-2-hydroxy-phenyl)-1,2,4-trimethyl-H
pyrrole-3-carboxylic acid ethyl ester hydrobromide 6h (180 mg, 0.66
mmol) was dissolved in 2.2 mL of hydrochloric acid (1 N) upon
cooling by an ice-water bath, followed by dropwise addition of 0.8
mL of aqueous sodium nitrite (50 mg, 0.72 mmol). After the mixture
was stirred for 20 minutes,
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (126 mg, 0.59
mmol) was added. The mixture was adjusted to pH 8.about.9 with
saturated aqueous sodium bicarbonate. Then the generated bubbles
were quenched with 2 mL of ethanol. The reaction mixture was warmed
up to room temperature and reacted overnight. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered and the filter cake was dissolved in a
mixture of mL of chloromethane and 20 mL of water. After mixing
well, the mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid. The separated organic layer was washed with
saturated brine, dried over anhydrous sodium sulfate, filtered to
remove the drying agent and concentrated under reduced pressure.
The resulting residue was purified by silica gel column
chromatography to obtain the title compound
5-{2-hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-yli-
dene)-hydrazino]-phenyl}-1,2,4-trimethyl-1H-pyrrole-3-carboxylic
acid ethyl ester 48 (80 mg, yield 25.8%) as a red solid.
[0439] MS m/z (ESI): 514.0 [M+1]
[0440] .sup.1H NMR (400 MHz, DMSO-d): .delta. 1.30 (m, 3H), 2.01
(m, 2H), 2.23 (s, 3H), 2.50 (s, 3H), 2.95 (m, 4H), 3.32 (s, 3H),
4.33 (m, 2H), 6.63 (m, 1H), 7.00 (m, 1H), 7.12 (m, 1H), 7.28 (m,
1H), 7.73 (m, 2H), 13.91 (br, 1H)
Example 49
5-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(1,1,3,3-tetramethyl-indan-5-yl)
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-thiophene-2-carboxylic
acid
##STR00136##
[0442] 5-(3-Amino-2-hydroxy-phenyl)-thiophene-2-carboxylic acid
hydrobromide 12c (260 mg, 0.823 mmol) was dissolved in 2.7 mL of
hydrochloric acid (1 N) upon cooling by an ice-water bath, followed
by dropwise addition of 1.1 mL of aqueous sodium nitrite (62 mg,
0.91 mmol). After the mixture was stirred for 20 minutes,
5-methyl-2-(1,1,3,3-tetramethyl-indan-5-yl)-2,4-dihydro-pyrazol-3-one
19d (200 mg, 0.74 mmol) was added. The mixture was adjusted to pH 8
with saturated aqueous sodium bicarbonate. Then the generated
bubbles were quenched with 2 mL of ethanol. The reaction mixture
was warmed up to room temperature and reacted for 3 hours. The
reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was filtered and 20 mL of water was
added to the filter cake. The mixture was adjusted to pH 3.about.4
with concentrated hydrochloric acid and filtered. The filter cake
was dried and purified by silica gel column chromatography to
obtain the title compound
5-(2-hydroxy-3-{N'-[3-methyl-5-oxo-1-(1,1,3,3-tetramethyl-indan-5-yl)-1,5-
-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-thiophene-2-carboxylic
acid 49 (216 mg, yield 56.5%) as a red solid.
[0443] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.30 (m, 12H),
1.93 (s, 2H), 2.34 (s, 3H), 7.19 (t, J=8.0 Hz, 1H), 7.25 (d, J=8.0
Hz, 1H), 7.57 (d, J=8.0 Hz, 1H), 7.66 (m, 2H), 7.71 (d, J=8.0 Hz,
1H), 7.77 (m, 2H), 10.10 (s, 1H), 13.06 (br, 1H), 13.72 (br,
1H)
Example 50
5-(2-Hydroxy-5-methyl-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthal-
en-2-yl)-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-furan-2-carboxy-
lic acid
##STR00137##
[0445] 5-(3-Amino-2-hydroxy-5-methyl-phenyl)-furan-2-carboxylic
acid hydrobromide 46c (120 mg, 0.38 mmol) was dissolved in 1.3 mL
of 1 N hydrochloric acid upon cooling by an ice-water bath,
followed by dropwise addition of 0.5 mL of aqueous sodium nitrite
(29 mg, 0.42 mmol). After the mixture was reacted for 20 minutes,
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
3i (78 mg, 0.34 mmol) was added. The mixture was adjusted to pH 8
with saturated aqueous sodium bicarbonate followed by addition of 2
mL of ethanol. The reaction mixture was reacted at room temperature
overnight. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was added to 20 mL of water. The mixture was
adjusted to pH 3.about.4 with concentrated hydrochloric acid. The
mixture was filtered and the filter cake was dried and purified by
silica gel column chromatography to obtain the title compound
5-(2-hydroxy-5-methyl-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydr-
o-naphthalen-2-yl)-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-furan-
-2-carboxylic acid 50 (56 mg, yield 34.8%) as a red solid.
[0446] MS m/z (ESI): 470.8 [M-1]
[0447] .sup.1H NMR (400 MHz, DMSO-d): .delta. 1.75 (m, 4H), 2.31
(s, 3H), 2.37 (s, 3H), 2.73 (m, 4H), 7.13 (m, 2H), 7.35 (m, 2H),
7.50 (s, 1H), 7.62 (m, 2H)
Example 51
4-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-thiophene-2-carboxylic
acid
##STR00138##
[0448] Step 1
4-(3-Nitro-2-methoxy-phenyl)-thiophene-2-carboxylic acid
[0449]
2-(2-Methoxy-3-nitro-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolan-
e 6a (0.81 g, 2.9 mol), 4-bromo-thiophene-2-carboxylic acid (0.3 g,
1.45 mmol), tetrakis (triphenylphosphine)palladium (80 mg, 0.073
mmol) and sodium carbonate (0.31 g, 2.9 mmol) were dissolved in a
solvent mixture of 20 mL of 1,4-dioxane and 10 mL of water. The
reaction was heated to reflux for 0.5 hours. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was adjusted to pH 3 with 1 N hydrochloric acid and
extracted with ethyl acetate (20 mL.times.3). The combined organic
extracts were concentrated under reduced pressure and the residue
was purified by silica gel column chromatography to obtain the
title compound 4-(3-nitro-2-methoxy-phenyl)-thiophene-2-carboxylic
acid 51a (0.54 g) as a brown oil, which was directly used in the
next step.
[0450] MS m/z (ESI): 277.6 [M-1]
Step 2
4-(3-Amino-2-methoxy-phenyl)-thiophene-2-carboxylic acid
[0451] 4-(3-Nitro-2-methoxy-phenyl)-thiophene-2-carboxylic acid 51a
(400 mg, 1.45 mmol) was dissolved in 30 mL of ethyl acetate
followed by addition of 100 mg of palladium on carbon and ammonium
formate (360 mg, 5.8 mmol). The mixture was heated to reflux for 3
hours. The reaction was monitored by TLC until the disappearance of
the starting materials. The mixture was filtered to remove
palladium on carbon and concentrated under reduced pressure to
obtain the title compound
4-(3-amino-2-methoxy-phenyl)-thiophene-2-carboxylic acid 51b (410
mg) as a brown oil, which was directly used in the next step,
[0452] MS m/z (ESI): 247.8 [M-1]
Step 3
4-(3-Amino-2-hydroxy-phenyl)-thiophene-2-carboxylic acid
hydrobromide
[0453] 4-(3-Amino-2-methoxy-phenyl)-thiophene-2-carboxylic acid
hydrobrobromide 51b (360 mg, 1.45 mmol) was dissolved in 5 mL of
dichloromethane followed by dropwise addition of boron tribromide
(2.8 mL, 5.6 mmol). The reaction mixture was reacted at room
temperature for 4.5 hours. The reaction was monitored by TLC until
the disappearance of the starting materials and quenched with 5 mL
of methanol. The mixture was concentrated under reduced pressure.
The residue was diluted with 10 mL of ethyl acetate and stirred for
0.5 hours. The mixture was filtered and the filter cake was dried
to obtain the title compound
4-(3-amino-2-hydroxy-phenyl)-thiophene-2-carboxylic acid
hydrobromide 51c (80 mg, yield 17.5%) as a grey solid.
[0454] MS m/z (ESI): 236.1 [M+1]
Step 4
4-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-thiophene-2-carboxylic
acid
[0455] 4-(3-Amino-2-hydroxy-phenyl)-thiophene-2-carboxylic acid
hydrobromide 51c (80 mg, 0.25 mmol) was dissolved in 1 mL of 1 N
hydrochloric acid upon cooling by an ice-water bath, followed by
dropwise addition of 0.3 mL of aqueous sodium nitrite (19 mg, 0.28
mmol). After the mixture was reacted for 20 minutes,
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
3i (52 mg, 0.23 mmol) was added. The mixture was adjusted to pH 8
with saturated aqueous sodium bicarbonate followed by addition of 2
mL of ethanol. The reaction mixture was reacted overnight at room
temperature. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was added to 20 mL of water. The mixture was
adjusted to pH 3.about.4 with concentrated hydrochloric acid. The
mixture was filtered and 4 mL of ethyl acetate was added to the
filter cake. The resulting mixture was stirred for 2 hours and
filtered, the filter cake was dried to obtain the title compound
4-(2-hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)-
-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-thiophene-2-carboxylic
acid 51 (11 mg, yield 10.2%) as a black solid.
[0456] MS m/z (ESI): 472.8 [M-1]
[0457] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.76 (m, 4H),
2.33 (s, 3H), 2.74 (m, 4H), 7.13 (m, 2H), 7.33 (m, 1H), 7.65 (m,
3H), 8.06 (d, J=1.6 Hz, 1H), 8.14 (d, J=1.6 Hz, 1H), 9.68 (s, 1H),
13.75 (s, 1H)
Example 52
4-{2-Hydroxy-3-[N'-(1-indan-5-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylidene-
)-hydrazino]-phenyl}-thiophene-2-carboxylic acid
##STR00139##
[0459] 4-(3-Amino-2-hydroxy-phenyl)-thiophene-2-carboxylic acid
hydrobromide 51c (120 mg, 0.38 mmol) was dissolved in 2.7 mL of 1 N
hydrochloric acid upon cooling by an ice-water bath, followed by
dropwise addition of 0.45 mL of aqueous sodium nitrite (29 mg, 0.42
mmol). After the mixture was reacted for 20 minutes,
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (73 mg, 0.34
mmol) was added. The mixture was adjusted to pH 8 with saturated
aqueous sodium bicarbonate followed by addition of 2 mL of ethanol.
The reaction mixture was reacted overnight at room temperature. The
reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was filtered and the filter cake
was added to 20 mL of water. The mixture was adjusted to pH
3.about.4 with concentrated hydrochloric acid and filtered. Then 5
mL of ethyl acetate was added to the filter cake and the mixture
was stirred for 1 hour. The mixture was filtered and the filter
cake was dried to obtain the title compound
4-{2-hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-yli-
dene)-hydrazino]-phenyl}-thiophene-2-carboxylic acid 52 (45 mg,
yield 28.7%) as a yellow solid.
[0460] MS m/z (ESI): 458.8 [M-1]
[0461] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.05 (m, 2H),
2.32 (s, 3H), 2.87 (m, 4H), 7.13 (t, J=8.0 Hz, 1H), 7.32 (m, 2H),
7.67 (m, 2H), 7.78 (s, 1H), 8.05 (d, J=1.6 Hz, 1H), 8.13 (d, J=1.2
Hz, 1H), 9.68 (s, 1H), 13.79 (s, 1H)
Example 53
5-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-2-methyl-furan-3-carboxy-
lic acid
##STR00140## ##STR00141##
[0462] Step 1
5-Bromo-2-methyl-furan-3-carboxylic acid methyl ester
[0463] 2-Methyl-furan-3-carboxylic acid methyl ester (2.0 g, 14.3
mmol) was dissolved in toluene followed by addition of
2,2'-azobis(2-methylpropionitrile) (10 mg, 0.06 mmol). The mixture
was cooled to 0.degree. C. by an ice-water bath followed by
addition of N-bromosuccinimide (2.8 g, 15.7 mmol). Upon completion
of the addition, the ice-water bath was removed and the reaction
mixture was stirred at room temperature overnight. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered and the filtrate was concentrated under
reduced pressure. The residue was purified by silica gel column
chromatography to obtain the title compound
5-bromo-2-methyl-furan-3-carboxylic acid methyl ester 53a (1.9 g,
yield 61%) as a colourless oil.
Step 2
5-(2-Methoxy-3-nitro-phenyl)-2-methyl-furan-3-carboxylic acid
methyl ester
[0464] 5-Bromo-2-methyl-furan-3-carboxylic acid methyl ester 53a
(0.65 g, 3.0 mmol) and
2-(2-methoxy-3-nitro-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
6a (1.0 g, 3.58 mmol) were dissolved in 1,4-dioxane (15 mL)
followed by addition of tetrakis (triphenylphosphine)palladium (173
mg, 0.15 mmol) and sodium carbonate (636 mg, 6.0 mmol). The
reaction mixture was heated to reflux at 100.degree. C. for 3
hours. The reaction was monitored by TLC until the disappearance of
the starting materials. The mixture was cooled and filtered. The
filtrate was concentrated under reduced pressure and the residue
was purified by silica gel column chromatography to obtain the
title compound
5-(2-methoxy-3-nitro-phenyl)-2-methyl-furan-3-carboxylic acid
methyl ester 53b (659 mg, yield 75%) as a white solid.
Step 3
5-(2-Methoxy-3-nitro-phenyl)-2-methyl-furan-3-carboxylic acid
[0465] 5-(2-Methoxy-3-nitro-phenyl)-2-methyl-furan-3-carboxylic
acid methyl ester 53b (650 mg, 2.23 mmol) was dissolved in
methanol, followed by addition of sodium hydroxide (268 mg, 6.7
mmol). Upon completion of the addition, the reaction mixture was
stirred at 50.degree. C. for 3 hours. The reaction was monitored by
TLC until the disappearance of the starting materials. The mixture
was concentrated under reduced pressure and adjusted to pH
3.about.4 with 1 N hydrochloric acid to form a copious amount of
precipitates. The mixture was filtered and the filter cake was
recrystallized from a solvent mixture of hexane/ethyl acetate
(V:V=5:1) to obtain the title compound
5-(2-methoxy-3-nitro-phenyl)-2-methyl-furan-3-carboxylic acid 53c
(450 mg, yield 84%) as a white solid.
[0466] MS m/z (ESI): 275.7 [M-1]
[0467] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 12.80 (s, 1H),
8.03 (dd, J=8.0 Hz, 1H), 7.87 (dd, J=8.0 Hz, 1H), 7.44 (t, J=8.0
Hz, 1H), 7.13 (s, 1H), 3.83 (s, 3H), 2.64 (s, 3H)
Step 4
5-(2-Methoxy-3-amino-phenyl)-2-methyl-furan-3-carboxylic acid
[0468] 5-(2-Methoxy-3-nitro-phenyl)-2-methyl-furan-3-carboxylic
acid 53c (450 mg, 1.62 mmol) was dissolved in methanol, followed by
addition of 45 mg of palladium on carbon. The reaction mixture was
heated to reflux for 4 hours under hydrogen atmosphere. The
reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was filtered and the filtrate was
concentrated under reduced pressure to obtain the title compound
5-(2-methoxy-3-amino-phenyl)-2-methyl-furan-3-carboxylic acid 53d
(370 mg, yield 92%) as a white solid.
[0469] MS m/z (ESI): 245.8 [M-1]
[0470] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 6.97 (s, 1H),
6.87 (m, 2H), 6.68 (m, 1H), 5.06 (br, 2H), 3.63 (s, 3H), 2.59 (s,
3H)
Step 5
5-(2-Hydroxy-3-amino-phenyl)-2-methyl-furan-3-carboxylic acid
hydrobromide
[0471] 5-(2-Methoxy-3-amino-phenyl)-2-methyl-furan-3-carboxylic
acid 53d (370 mg, 1.5 mmol) was dissolved in dichloromethane. The
mixture was cooled to 0.degree. C. by an ice-water bath followed by
dropwise addition of a solution of boron tribromide in
dichloromethane (1 N, 3.6 mL). Upon completion of the addition, the
reaction mixture was reacted at room temperature for 2 hours. The
reaction was monitored by TLC until the disappearance of the
starting materials and quenched by addition of 0.5 mL of methanol.
The mixture was stirred for 30 minutes and concentrated under
reduced pressure. The resulting residue was diluted with 10 mL of
ethyl acetate and stirred for 30 minutes. The mixture was filtered
and the filter cake was dried to obtain the title compound
5-(2-hydroxy-3-amino-phenyl)-2-methyl-furan-3-carboxylic acid
hydrobromide 53e (240 mg, yield 46%) as a grey solid.
[0472] MS m/z (ESI): 231.7 [M-1]
[0473] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 7.57 (dd, J=8.0,
1H), 7.24 (dd, J=8.0, 1H), 7.05 (t, J==8.0, 1H), 2.61 (s, 3H)
Step 6
5-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-2-methyl-furan-3-carboxy-
lic acid
[0474] 5-(2-Hydroxy-3-amino-phenyl)-2-methyl-furan-3-carboxylic
acid hydrobromide 53e (200 mg, 0.64 mmol) was dissolved in 2.2 mL
of 1 N hydrochloric acid upon cooling by an ice-water bath,
followed by dropwise addition of 0.9 mL of aqueous sodium nitrite
(48 mg, 0.7 mmol). After the mixture was reacted for 20 minutes,
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
3i (131 mg, 0.57 mmol) was added. The mixture was adjusted to pH 8
with saturated aqueous sodium bicarbonate followed by addition of 2
mL ethanol. The reaction mixture was reacted overnight at room
temperature. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was filtered
and the filter cake was added to 20 mL of water. The mixture was
adjusted to pH 3.about.4 with concentrated hydrochloric acid. The
mixture was filtered and 8 mL of ethyl acetate was added to the
filter cake. After stirring for 1 hour, the mixture was filtered
and the filter cake was dried to obtain the title compound
5-(2-hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)-
-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-2-methyl-furan-3-carbox-
ylic acid 53 (200 mg, yield 73.8%) as a red solid.
[0475] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.75 (m, 4H),
2.31 (s, 3H), 2.62 (s, 3H), 2.77 (m, 4H), 7.14 (m, 3H), 7.47 (d,
J=7.6 Hz, 1H), 7.65 (m, 3H), 9.79 (s, 1H), 12.73 (br, 1H), 13.76
(br, 1H)
Example 54
5-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-furan-2-carboxylic
acid methyl ester
##STR00142##
[0476] Step 1
5-(2-Methoxy-3-nitro-phenyl)-furan-2-carboxylic acid methyl
ester
[0477]
2-(2-Methoxy-3-nitro-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolan-
e 6a (3.6 g, 12 mmol), 5-bromofuran-2-carboxylic acid methyl ester
(2.05 g, 10 mmol), tetrakis (triphenylphosphine)palladium (1.55 g,
0.5 mmol) and sodium carbonate (2.12 g, 20 mmol) were dissolved in
1,4-dioxane. The reaction mixture was heated to reflux for 3 hours.
The reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was filtered and the filtrate was
concentrated under reduced pressure. The residue was diluted with
30 mL of water and 50 mL of ethyl acetate. The separated organic
layer was concentrated under reduced pressure and recrystallized
from a solvent mixture of ethyl acetate and n-hexane to obtain the
title compound 5-(2-methoxy-3-nitro-phenyl)-furan-2-carboxylic acid
methyl ester 54a (500 mg, yield 18%) as a yellow solid.
[0478] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 8.23 (dd, J=7.6
Hz, 1H), 7.83 (dd, J=7.6, 1H), 7.35 (m, 2H), 7.16 (d, J=4.0 Hz,
1H), 3.98 (s, 3H), 3.94 (s, 3H)
Step 2
5-(3-Amino-2-methoxy-phenyl)-furan-2-carboxylic acid methyl
ester
[0479] 5-(2-Methoxy-3-nitro-phenyl)-furan-2-carboxylic acid methyl
ester 54a (500 mg, 1.8 mmol) was dissolved in methanol followed by
addition of 50 mg of palladium on carbon. The reaction mixture was
heated to reflux for 4 hours under hydrogen atmosphere. The
reaction was monitored by TLC until the disappearance of the
starting materials. The mixture was concentrated under reduced
pressure. The resulting residue was recrystallized from a solvent
mixture of ethyl acetate/n-hexane (V:V=1:5) and dried to obtain the
title compound 5-(3-amino-2-methoxy-phenyl)-furan-2-carboxylic acid
methyl ester 54b (370 mg, yield 83%) as a white solid.
Step 3
5-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid methyl
ester
[0480] 5-(3-Amino-2-methoxy-phenyl)-furan-2-carboxylic acid methyl
ester 54b (350 mg, 1.42 mmol) was dissolved in dichloromethane
followed by dropwise addition of boron tribromide (3.3 mL, 2.0
mol/L). The reaction mixture was stirred at room temperature for 2
hours. The reaction was monitored by TLC until the disappearance of
the starting materials and quenched with methanol. The mixture was
adjusted to pH 5.about.6 with saturated aqueous sodium bicarbonate
and extracted with ethyl acetate. The organic layers were
concentrated and then the resulting residue was purified by silica
gel column chromatography to obtain the title compound
5-(3-amino-2-hydroxy-phenyl)-furan-2-carboxylic acid methyl ester
54c (170 mg, yield 45.1%) as a grey solid.
[0481] MS m/z (ESI): 232.0 [M-1]
[0482] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 7.32 (d, J=7.6
Hz, 1H), 7.05 (d, J==7.6 Hz, 1H), 6.82 (m, 3H), 3.96 (s, 3H)
Step 4
5-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-furan-2-carboxylic
acid methyl ester
[0483] 5-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid methyl
ester 54c (110 mg, 0.47 mmol) was dissolved in hydrochloric acid
(1.6 mL, 1 mol/L) upon cooling by an ice-water bath, followed by
dropwise addition of 0.6 mL of aqueous sodium nitrite (36 mg, 0.52
mmol). After the mixture was reacted for 10 minutes,
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
3i (97 mg, 0.43 mmol) was added. The mixture was adjusted to pH
8.about.9 with saturated aqueous sodium bicarbonate. The reaction
mixture was reacted at room temperature for 24 hours. The reaction
was monitored by TLC until the disappearance of the starting
materials. The mixture was filtered and the filter cake was
dissolved in 1.5 mL of water. The mixture was adjusted to pH
3.about.4 with concentrated hydrochloric acid upon cooling by an
ice-water bath. The mixture was filtered and the filter cake was
purified by silica gel column chromatography to obtain the title
compound
5-(2-hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)-
-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-furan-2-carboxylic
acid methyl ester 54 (48 mg, yield 23.9%) as a red solid.
[0484] MS m/z (ESI): 470.7 [M-1]
[0485] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 1.76 (m, 4H),
2.31 (s, 3H), 2.73 (m, 4H), 3.86 (s, 3H), 7.12 (m, 1H), 7.17 (m,
1H), 7.22 (t, J=8.0 Hz, 1H), 7.46 (m, 1H), 7.56 (m, 1H), 7.65 (m,
2H), 7.72 (m, 1H), 10.02 (s, 1H), 13.72 (br, 1H)
Example 55
5-{2-Hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylid-
ene)-hydrazino]-phenyl}-furan-2-carboxylic acid methyl ester
##STR00143##
[0487] 5-(3-Amino-2-hydroxy-phenyl)-furan-2-carboxylic acid methyl
ester 54c (110 mg, 0.47 mmol) was dissolved in hydrochloric acid
(1.6 mL, 1 mol/L) upon cooling by an ice-water bath, followed by
dropwise addition of 0.6 mL of aqueous sodium nitrite (36 mg, 0.52
mmol). After the mixture was reacted for 20 minutes,
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (91 mg, 0.43
mmol) was added. The mixture was adjusted to pH 8.about.9 with
saturated aqueous sodium bicarbonate. The reaction mixture was
reacted at room temperature for 24 hours. The reaction was
monitored by TLC until the disappearance of the starting materials
and quenched with ethanol. The mixture was filtered and the filter
cake was dissolved in 15 mL of water. The mixture was adjusted to
pH 3.about.4 with concentrated hydrochloric acid upon cooling by an
ice-water bath. The mixture was filtered and the filter cake was
purified by silica gel column chromatography to obtain the title
compound
5-{2-hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyra-
zol-4-ylidene)-hydrazino]-phenyl}-furan-2-carboxylic acid methyl
ester 55 (137 mg, yield 70.3%) as a red solid.
[0488] MS m/z (ESI): 456.7 [M-1]
[0489] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 2.05 (m, 2H),
2.33 (s, 3H), 2.89 (m, 4H), 3.86 (s, 3H), 7.18 (m, 2H), 7.30 (d,
J=8.4 Hz, 1H), 7.46 (d, J=3.6 Hz, 1H), 7.56 (m, 1H), 7.71 (m, 2H),
7.78 (s, 1H)
Example 56
3'-{N'-[1-(2,2-Dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-y-
lidene]-hydrazino}-2'-hydroxy-biphenyl-3-carboxylic acid
##STR00144##
[0491] 3'-Amino-2'-hydroxy-biphenyl-3-carboxylic acid hydrobromide
1f (150 mg, 0.5 mmol) was dissolved in hydrochloric acid (1.7 mL, 1
mol/L) upon cooling by an ice-water bath, followed by dropwise
addition of 0.6 mL of aqueous sodium nitrite (38 mg, 0.55 mmol).
After the mixture was reacted for 20 minutes,
2-(2,2-dimethyl-indan-5-yl)-5-methyl-2,4-dihydro-pyrazol-3-one 45d
(109 mg, 0.45 mmol) was added. The mixture was adjusted to pH
8.about.9 with saturated aqueous sodium bicarbonate. Then the
generated bubbles were quenched with ethanol. The reaction mixture
was reacted overnight at room temperature. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered and the filter cake was added to 15 mL of
water. The mixture was adjusted to pH 3.about.4 with concentrated
hydrochloric acid and filtered. The filter cake was washed with
ethyl acetate (1 mL.times.3) and then dried to obtain the title
compound
3'-{N'-[1-(2,2-dimethyl-indan-5-yl)-3-methyl-5-oxo-1,5-dihydro-p-
yrazol-4-ylidene]-hydrazino}-2'-hydroxy-biphenyl-3-carboxylic acid
56 (16 mg, yield 7.6%) as a yellow solid.
[0492] MS m/z (ESI): 480.7 [M-1]
[0493] .sup.1H NMR (400 MHz, DMSO-d.sub.6): 1.13 (s, 6H), 2.32 (s,
3H), 2.70 (m, 4H), 7.14 (m, 2H), 7.22 (d, J=8.8 Hz, 1H), 7.61 (t,
J=8.0 Hz, 1H), 7.66 (d, J=7.6 Hz, 1H), 7.70 (m, 2H), 7.80 (d, J=7.6
Hz, 1H), 7.95 (d, J=8.0 Hz, 1H), 8.13 (s, 1H)
Example 57
4-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-1H-pyrrole-2-carboxylic
acid
##STR00145## ##STR00146##
[0494] Step 1
2,2,2-Trichloro-1-(1H-pyrrol-2-yl)-ethanone
[0495] Trichloroacetyl chloride (45 g, 247 mmol) was dissolved in
100 mL of ether followed by dropwise addition of a solution of
1H-Pyrrole (15.4 g, 230 mmol) in 100 mL of ether a 200 mL of
aqueous potassium carbonate (20 g, 145 mol). Upon completion of the
addition, the reaction mixture was stirred at room temperature for
1 hour. The reaction was monitored by TLC until the disappearance
of the starting materials. The layers were separated and the
organic layer was dried over anhydrous magnesium sulfate, filtered
to remove the drying agent and concentrated under reduced pressure.
The residue was purified by silica gel column chromatography to
obtain the title compound
2,2,2-trichloro-1-(1H-pyrrol-2-yl)-ethanone 57b (38 g, yield 778%)
as a white solid.
[0496] MS m/z (ESI): 210.3 [M-1]
Step 2
2,2,2-Trichloro-1-(4-iodo-1H-pyrrol-2-yl)ethanone
[0497] 2,2,2-Trichloro-1-(1H-pyrrol-2-yl)-ethanone 57b (32 g, 151.8
mmol) was dissolved in 250 mL of dichloromethane followed by
dropwise addition of a solution of iodine monochloride (25 g, 153
mmol) in 125 mL of dichloromethane. Upon completion of the
addition, the reaction mixture was stirred at room temperature for
2 hours. The reaction was monitored by TLC until the disappearance
of the starting materials. The mixture was washed with saturated
aqueous sodium carbonate, aqueous sodium thiosulfate (2 M) and
saturated brine successively, dried over anhydrous magnesium
sulfate, filtered to remove the drying agent and concentrated under
reduced pressure. The residue was purified by silica gel column
chromatography to obtain the title compound
2,2,2-trichloro-1-(4-iodo-1H-pyrrol-2-yl)-ethanone 57c (47 g, yield
92%) as a yellow solid.
[0498] MS m/z (ESI): 336.4 [M-1]
Step 3
4-Iodo-1H-pyrrole-2-carboxylic acid methyl ester
[0499] 2,2,2-Trichloro-1-(4-iodo-1H-pyrrol-2-yl)-ethanone 57c (47
g, 136 mmol) was dissolved in 265 mL of methanol followed by
dropwise addition of a solution of sodium methoxide (17.23 g, 163
mmol) in 200 mL of methanol. The reaction mixture was stirred at
room temperature for 1 hour. The reaction was monitored by TLC
until the disappearance of the starting materials. The mixture was
concentrated under reduced pressure. The residue was diluted with
20 mL of water and then extracted with ethyl acetate (30
mL.times.3). The combined organic extracts were washed with
saturated brine, dried over anhydrous sodium sulfate, filtered to
remove the drying agent and concentrated under reduced pressure to
obtain the title compound 4-iodo-1H-pyrrole-2-carboxylic acid
methyl ester 57d (32.2 g, yield 92.5%) as a grey solid.
[0500] MS m/z (ESI): 250.1 [M-1]
Step 4
4-Iodo-1-(toluene-4-sulfonyl)-1H-pyrrole-2-carboxylic acid methyl
ester
[0501] 4-Iodo-1H-pyrrole-2-carboxylic acid methyl ester 57d (25.1
g, 100 mmol) was dissolved in 150 mL of dichloromethane followed by
addition of triethylamine (30.6 mL, 220 mmol),
4-dimethylaminopyridine (1.22 g, 10 mmol) and p-toluenesulfonic
acid (21 g, 110 mmol). The reaction mixture was reacted at
20.degree. C. overnight. The reaction was monitored by TLC until
the disappearance of the starting materials and quenched by
addition of 30 mL of hydrochloric acid (1 N). The mixture was
extracted with dichloromethane (50 mL.times.3). The combined
organic extracts were washed with saturated aqueous sodium
carbonate and saturated brine successively, dried over anhydrous
sodium sulfate, filtered to remove the drying agent and
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography to obtain the title compound
4-iodo-1-(toluene-4-sulfonyl)-1H-pyrrole-2-carboxylic acid methyl
ester 57e (32.5 g, yield 80.2%) as a white solid.
[0502] MS m/z (ESI): 405.8 [M+1]
Step 5
4-(3-Nitro-2-methoxy-phenyl)-1-(p-tolylsulfonyl)-1H-pyrrole-2-carboxylic
acid methyl ester
[0503] To a solution of 15 mL of 1,4-dioxane and 5 mL of water was
added
2-(2-methoxy-3-nitro-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
6a (2.05 g, 5.5 mmol) followed by
4-iodo-1-(toluene-4-sulfonyl)-1H-pyrrole-2-carboxylic acid methyl
ester 57e (2.03 g, 5 mmol), potassium carbonate (1.38 g, 10 mmol)
and tetrakis(triphenylphosphine)palladium (144 mg, 0.125 mmol).
Upon completion of the addition, the reaction mixture was reacted
at 80.degree. C. for 30 minutes under microwave. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was concentrated under reduced pressure and the residue
was diluted with 20 mL of water. After mixing well, the mixture was
extracted with ethyl acetate (20 mL.times.3). The combined organic
extracts were washed with saturated brine, dried over anhydrous
magnesium sulfate, filtered to remove the drying agent and
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography to obtain the title compound
4-(3-nitro-2-methoxy-phenyl)-1-(p-tolylsulfonyl)-1H-pyrrole-2-carboxylic
acid methyl ester 57f (1.04 g, yield 48%) as a grey solid.
[0504] MS m/z (ESI): 431.0 [M+-1]
[0505] .sup.1H NMR (400 MHz, CDCl.sub.3): .delta. 8.225.about.8.231
(m, 1H), 7.979.about.8.00 (m, 2H), 7.710.about.7.765 (m, 1H),
7.452.about.7.457 (m, 1H), 7.389.about.7.409 (m, 2H),
7.271.about.7.311 (m, 2H), 3.839 (s, 3H), 3.829 (s, 2H), 2.488 (s,
3H)
Step 6
4-(3-Nitro-2-methoxy-phenyl)-1H-pyrrole-2-carboxylic acid
[0506]
4-(3-Nitro-2-methoxy-phenyl)-1-(p-tolylsulfonyl)-1H-pyrrole-2-carbo-
xylic acid methyl ester 57f (1.04 g, 2.42 mmol) and lithium
hydroxide monohydrate (1.01 g, 24.19 mmol) were added to a solvent
mixture of 10 mL of N,N-dimethylformamide and 5 mL, of water. The
reaction mixture was reacted at 100.degree. C. for 30 minutes under
microwave. The reaction was monitored by TLC until the
disappearance of the starting materials. The mixture was adjusted
to pH 3 with hydrochloric acid (1 N) to form a lot of precipitates.
The mixture was filtered and the filter cake was dried to obtain
the title compound
4-(3-nitro-2-methoxy-phenyl)-1H-pyrrole-2-carboxylic acid 57g (350
mg, yield 50%) as a yellow solid.
[0507] MS m/z (ESI): 260.8 [M-1]
Step 7
4-(3-Amino-2-methoxy-phenyl)-1H-pyrrole-2-carboxylic acid
[0508] 4-(3-Nitro-2-methoxy-phenyl)-1H-pyrrole-2-carboxylic acid
57g (633 mg, 2.41 mmol) was dissolved in 15 mL of ethyl acetate
followed by addition of 127 mg of palladium on carbon and ammonium
formate (609 mg, 9.66 mmol). Upon completion of the addition, the
reaction mixture was heated to reflux for 2 hours. The reaction was
monitored by TLC until the disappearance of the starting materials.
The mixture was filtered to remove palladium on carbon and the
filtrate was concentrated under reduced pressure. The residue was
purified by silica gel column chromatography to obtain the title
compound 4-(3-amino-2-methoxy-phenyl)-1H-pyrrole-2-carboxylic acid
57h (130 mg, yield 23.2%) as a grey solid.
[0509] MS m/z (ESI): 230.8 [M-1]
[0510] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 11.631 (s, 1H),
7.305 (s, 1H), 7.025 (s, 1H), 6.712.about.6.798 (m, 2H),
6.524.about.6.543 (m, 1H), 3.514 (s, 3H)
Step 8
4-(3-Amino-2-hydroxy-phenyl)-1H-pyrrole-2-carboxylic acid
[0511] To a solution of
4-(3-amino-2-methoxy-phenyl)-1H-pyrrole-2-carboxylic acid 57h (130
mg, 0.56 mmol) in 2 mL of dichloromethane was added boron
tribromide (1.12 mL, 2.24 mmol). The reaction mixture was stirred
at room temperature for 6 hours. The reaction was monitored by TLC
until the disappearance of the starting materials and quenched with
methanol. The mixture was concentrated under reduced pressure and
the residue was purified by silica gel column chromatography to
obtain the title compound
4-(3-amino-2-hydroxy-phenyl)-1H-pyrrole-2-carboxylic acid 57i (140
mg, yield 99%) as a grey solid.
[0512] .sup.1H NMR (400 MHz, CD.sub.3OD): .delta. 7.312 (s, 1H),
7.178 (s, 1H), 7.006.about.7.028 (m, 1H), 6.82.about.6.837 (m,
2H)
Step 9
4-(2-Hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)--
1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-1H-pyrrole-2-carboxylic
acid
[0513] 4-(3-Amino-2-hydroxy-phenyl)-1-pyrrole-2-carboxylic acid 57i
(130 mg, 0.43 mmol) was dissolved in hydrochloric acid (1.5 mL, 1
mol/L) upon cooling by an ice-water bath, followed by dropwise
addition of 0.6 mL of aqueous sodium nitrite (33 mg, 0.47 mmol).
After the mixture was reacted for 10 minutes,
5-methyl-2-(5,6,7,8-tetrahydro-naphthalen-2-yl)-2,4-dihydro-pyrazol-3-one
3i (89 mg, 0.3 mmol) was added. The mixture was adjusted to pH
8.about.9 with saturated aqueous sodium bicarbonate. The reaction
mixture was reacted at room temperature for 24 hours. The reaction
was monitored by TLC until the disappearance of the starting
materials. The mixture was filtered and the filter cake was
dissolved in 15 mL of water. The mixture was adjusted to pH
3.about.4 with concentrated hydrochloric acid upon cooling by an
ice-water bath. The mixture was filtered and the filter cake was
washed with ethyl acetate and dried to obtain the title compound
4-(2-hydroxy-3-{N'-[3-methyl-5-oxo-1-(5,6,7,8-tetrahydro-naphthalen-2-yl)-
-1,5-dihydro-pyrazol-4-ylidene]-hydrazino}-phenyl)-1H-pyrrole-2-carboxylic
acid 57 (38 mg, yield 21.3%) as a grey solid. .sup.1H NMR (400 MHz,
DMSO-d.sub.6): .delta. 11.74 (1H, br), 11.64 (1H, br), 7.60 (1H, d,
J=8.2 Hz), 7.49 (1H, d, J=8.0 Hz), 7.32 (3H, m), 7.05 (3H, m), 2.67
(4H, m), 1.95 (3H, s), 1.13 (4H, m)
Example 58
4-{2-Hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-ylid-
ene)-hydrazino]-phenyl}-1H-pyrrole-2-carboxylic acid
##STR00147##
[0515] 4-(3-Amino-2-hydroxy-phenyl)-1H-pyrrole-2-carboxylic acid
57i (240 mg, 0.8 mmol) was dissolved in hydrochloric acid (3 mL, 1
mol/L) upon cooling by an ice-water bath, followed by dropwise
addition of 1.1 mL of aqueous sodium nitrite (61 mg, 0.88 mmol).
After the mixture was reacted for 20 minutes,
2-indan-5-yl-5-methyl-2,4-dihydro-pyrazol-3-one 1i (154 mg, 0.72
mmol) was added. The mixture was adjusted to pH 8.about.9 with
saturated aqueous sodium bicarbonate. The reaction mixture was
reacted at room temperature for 24 hours. The reaction was
monitored by TLC until the disappearance of the starting materials
and quenched with ethanol. The mixture was filtered and the filter
cake was dissolved in 15 mL of water. The mixture was adjusted to
pH 3.about.4 with concentrated hydrochloric acid upon cooling by an
ice-water bath. The mixture was filtered and the filter cake was
purified by silica gel column chromatography to obtain the title
compound
4-{2-hydroxy-3-[N'-(1-indan-5-yl-3-methyl-5-oxo-1,5-dihydro-pyrazol-4-yli-
dene)-hydrazino]-phenyl}-1H-pyrrole-2-carboxylic acid 58 (101 mg,
yield 28.4%) as a red solid.
[0516] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. 11.83 (1H, br),
11.77 (1H, br), 7.57 (1H, d, J=7.2 Hz), 7.50 (1H, s), 7.37 (4H, m),
7.26 (1H, m), 7.10 (1H, m), 2.87 (4H, m), 2.32 (3H, s), 2.04 (2H,
m)
[0517] Biological Assay
Test Example 1
[0518] pro-proliferation effect of a series of TPO compounds on
BAF3-TPOR cell.
[0519] 1. Material and Reagents.
[0520] a) RPMI Medium 1640, powder, 10*1 L, containing HEPES (Gibco
Catalog No. 23400021).
[0521] b) Fetal Bovine Serum (Gibco Catalog No. 10099-141).
[0522] c) PENICILLIN STREPTOMYCIN SOL (Gibco Catalog No.
15140-122).
[0523] d) Geneticin (G418) (Gibco Catalog No. 11811-098),
[0524] e) Recombinant mouse IL-3 (chemicon Catalog No. IL015).
[0525] f) Human Thrombopoietin R Mab (TPO) (R&D Catalog No.
MAB1016).
[0526] g) DMSO, (AppliChem Catalog No. A3672).
[0527] h) QuikChange.RTM. Multi Site Directed Mutagenesis Kit, 10
Runs (Stratagene ST200515).
[0528] i) Cell Counting Kit-8 (Dojindo, Catalog No. CK04-13)
[0529] j) BaF3 cell (Union cell culture center, Catalog No.
0095)
[0530] k) EX-EGFP-M02 (FulenGen Catalog No. EX-EGFP-M02
Control)
[0531] l) EX-B0010-M02 (FulenGen Catalog No. EX-B0010-M02)
[0532] 2. Operating Process:
[0533] (1) Plasmid constructs: based on the TPOR sequence
information from Entrez (Gene ID: 4325, Refseq: NM.sub.--005373),
dual-site mutation was performed on EX-B0010-M02 plasmid by using
QuikChange.RTM. Multi Site Directed Mutagenesis Kit (Stratagene).
The sequence of primers which contains multi-sites mutation was
designed as follows:
TABLE-US-00003 g491a: 5'-gggaacttcagatcagctgggaggagccg-3'
g491a_antisense: 5'-cggctcctcccagctgatctgaagttccc-3'; c965t:
5'-caggaccatgctagctcccaaggcttcttct-3', c965t_antisense:
5'-agaagaagccttgggagctagcatggtcctg-3'.
[0534] The E. coli.DH5.alpha. competent cell was transformed with
mutated plasmid, and positive colonies were picked up through
ampicillin selection. The mutation result was confirmed by sequence
analysis.
[0535] (2) BAF3-TPOR stable transfected cell line: the following
method was used to construct the BaF3 cell which stably
over-expressed functional human TPO receptor. The successfully
mutated EX-B0010-M02 plasmid (25 .mu.g) which expressed human TPO
receptor and screening gene neomycin was transfected into wild-type
BaF3 cells (1.times.10.sup.7) by electroporation at 250V for 18 ms
using an electric pulse generator (Electro Square Porator ECM830,
BTX Division of Genetronic, Inc. US). The stable transfected cells
BAF3-TPOR were selected with G418 (Gibco, US), then incubated with
RPMI1640 medium plus 10% FBS (Gibco, US), 800 ng/mL G418, 5 ng/mL,
rmIL-3 (Chemicon, US).
[0536] 3. Screening Compounds Assay
[0537] (1) Washing cells by centrifugation: a suitable amount of
cell suspension was centrifuged at 1000 rpm for 5 minutes, and the
supernatant was discarded, 10 mL of cell culture media without
containing IL-3 was added. Then the resulting cell suspension was
centrifuged at 1000 rpm for 5 minutes, and the supernatant was
discarded.
[0538] (2) 1 mL of cell culture media without containing IL-3 was
added to beat upon them to equality, and the number of a suitable
amount of cell suspension was counted after dilution.
[0539] (3) According to the result of the cell counting, a cell
suspension in a concentration of 100,000 cell/mL was prepared.
[0540] (4) 100 .mu.L of cell suspension was transferred to each
well of 96-well culture plate, and there were 3 parallel wells,
that is, there were a blank control group (B), a negative control
group (N), a positive control group of TPO(P) and a test compound
group (S).
[0541] (5) The test compound was dissolved in DMSO to prepare a 10
mM stock solution, and then the solution was diluted with RPMI 1640
medium into a series of test samples at different concentration: 30
.mu.M, 10 .mu.M, 3 .mu.M, 1 .mu.M, 0.3 .mu.M, 0.1 .mu.M, 0.03
.mu.M, 0.01 .mu.M, 0.003 .mu.M, 0.001 .mu.M.
[0542] (6) 10 .mu.L of test compound solution was transferred to
each well respectively; 1 .mu.L of rhTPO (10 .mu.g/mL) was added to
positive control well.
[0543] (7) The plates were incubated in an incubator at 5% CO.sub.2
and 37.degree. C. for 24 hours.
[0544] (8) After incubation, 10 .mu.L of CCK-8 solution was added
to each well and the plates were incubated in an incubator for 24
hours.
[0545] (9) OD value was detected at 450 nm by VICTOR3 (Perkin Elmer
1420-120) plate reader.
[0546] 4. Analytical Calculation
[0547] (1) The proliferation rate was calculated as follows:
Rate=[(S-B)/(P-B)]*100%
[0548] S: OD value of wells which contains test compound.
[0549] B: OD value of blank control wells
[0550] P: OD value of positive control wells
[0551] (2) The EC.sub.50 value was calculated by Origin 7.0
software.
[0552] 5. Results:
[0553] EC.sub.50 of TPO activity of the compounds of the present
disclosure
TABLE-US-00004 Example No. EC.sub.50 (nM) Eltrombopag 299 1 200 3
310 4 283 5 354 7 265 9 100 11 257 13 141 15 21 16 160 20 89 22 15
25 60 28 42 29 124 31 27 37 90 43 32 44 50 45 130 46 56 50 126 51
55 52 43 54 133 55 71 56 60
[0554] Pharmacokinetics Assay
Test Example 1
Pharmacokinetics Assay of the Compounds of the Present
Disclosure
[0555] 1. Purpose
[0556] The compounds of Example 1, Example 15 and Example 29 were
administrated intragastrically to Sprague-Dawley (SD) rats to
determine the drug concentration in plasma at different time
points. The pharmacokinetics of the compounds of the present
disclosure was studied and evaluated in rats.
[0557] 2. Protocol
[0558] 2.1 Samples
Compounds of Example 1, Example 15 and Example 29
[0559] 2.2 Experimental animals
[0560] 24 healthy adult SD rats, male and female in half, were
purchased from SINO-BRITISH SIPPR/BK LAB.ANIMAL LTD., CO, License
number: SCXK (Shanghai) 2003-0002
[0561] 2.3 Instrument
[0562] TSQ Quantum Ultra AM Triple Quadrupole Mass Spectrometer,
Thermo Finnigan Corp., USA;
[0563] Agilent 1200 high performance liquid chromatograph, Agilent
Corp., USA;
[0564] 2.3 Preparation of Test Compounds
[0565] The test compound was diluted with 1% sodium carboxymethyl
cellulose to 0.5 mg/mL (calculated as the free acid form) of
suspension before use.
[0566] 2.5 Administration
[0567] 24 healthy adult SD rats, male and female in half, were
divided into 5 groups. After an overnight fast, the rats were
administered intragastrically at a dose of 5.0 mg/kg (calculated as
the free acid form), at a volume of 10 mL/kg.
[0568] 2.6 Sample Collection
[0569] 24 healthy adult SD rats, male and female in half, were
administered intragastrically at a dose of 5.0 mg/kg after an
overnight fast. Blood samples (0.2 mL) were taken from eye socket
at pre administration and at 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0,
8.0, 11.0, 14.0, 24.0, 36.0 and 48.0 hours post administration,
which were stored in heparinized tubes and centrifuged for 10
minutes at 3,500 rpm. The plasma samples were stored at -20.degree.
C. until analysis.
[0570] 2.7 Analytical Methods
[0571] 50 .mu.L, of rat plasma, obtained at various time points
after administration, 20 .mu.L of internal standard solution and 20
.mu.L of a solvent mixture of methanol and water (80:20, v/v) were
mixed well, and then 150 .mu.L of methanol was added to result in
protein precipitation. Then the mixture was mixed for 1 minute
using a vortexer and centrifuged for 10 minutes at 13,000 rpm. 20
.mu.L of the supernatant was analyzed by LC/MS/MS.
[0572] 2.8 Preparation of Calibration Curve
[0573] 50 .mu.L of blank plasma was spiked with a series of
standard solutions to produce final concentrations of 1.0, 5.0,
25.0, 50.0, 100.0, 250.0, 500.0 ng/mL, followed by addition of 20
.mu.L of internal standard solution, which was treated according to
plasma sample pretreatment protocol. A typical calibration curve
equation was obtained with plasma concentration as the abscissa and
the chromatographic peak area ratio of sample to internal standard
as the ordinate, using weighted least squares method (w=1/x.sup.2)
for linear regression.
[0574] 2.9 Calculation of Pharmacokinetic Parameters
[0575] The compartmental model of pharmacokinetics was fitted for
the test compounds and the major pharmacokinetic parameters were
calculated in which C.sub.max and t.sub.max were the actually
measured values.
[0576] 3. Results of Pharmacokinetic Parameters
[0577] Pharmacokinetic Parameters of the compounds of the present
disclosure were shown as follows:
TABLE-US-00005 Pharmacokinetics Assay (5 mg/Kg) Plasma Mean
Apparent Conc. Curve Area Residence Distribution C.sub.max AUC
Half-Life Time Clearance Volume Number (.mu.g/mL) (.mu.g/mL *h)
t1/2(h) MRT(h) CL/F(l/h/kg) Vz/F(l/kg) Example 1 29.05 .+-. 11.44
131.99 .+-. 46.95 5.39 .+-. 0.94 4.96 .+-. 1.16 0.049 .+-. 0.039
0.35 .+-. 0.18 Example 15 6.63 .+-. 3.78 23.8 .+-. 17.26 3.47 .+-.
0.79 4.09 .+-. 0.99 0.28 .+-. 0.15 1.36 .+-. 0.70 Example 29 12.95
.+-. 5.96 44.78 .+-. 19.45 4.01 .+-. 0.63 4.29 .+-. 0.80 0.14 .+-.
0.088 0.77 .+-. 0.39 Eltrom- 8.47 .+-. 0.95 29.02 .+-. 3.82 6.11
.+-. 1.04 4.63 .+-. 1.03 0.18 .+-. 0.02 1.53 .+-. 0.26 bopag
[0578] The results of the current study indicate that the above
test compounds of the present disclosure are well absorbed after an
intragastric administration to rats.
Sequence CWU 1
1
4129DNAArtificial Sequenceg491a 1gggaacttca gatcagctgg gaggagccg
29229DNAArtificial Sequenceg491a antisense 2cggctcctcc cagctgatct
gaagttccc 29331DNAArtificial Sequencec965t 3caggaccatg ctagctccca
aggcttcttc t 31431DNAArtificial Sequencec965t antisense 4agaagaagcc
ttgggagcta gcatggtcct g 31
* * * * *