U.S. patent application number 15/723785 was filed with the patent office on 2018-01-25 for carboxylic acid compound, method for preparation thereof, and use thereof.
The applicant listed for this patent is INVENTISBIO SHANGHAI LTD.. Invention is credited to Yueheng Jiang.
Application Number | 20180022736 15/723785 |
Document ID | / |
Family ID | 55303876 |
Filed Date | 2018-01-25 |
United States Patent
Application |
20180022736 |
Kind Code |
A1 |
Jiang; Yueheng |
January 25, 2018 |
CARBOXYLIC ACID COMPOUND, METHOD FOR PREPARATION THEREOF, AND USE
THEREOF
Abstract
The present invention relates to the technical field of
medicine, and specifically relates to the carboxylic acid compound
represented by the chemical formula I or chemical formula II, and a
pharmaceutically acceptable salt, a prodrug, and a solvate thereof,
and a method for preparation thereof, as well as a pharmaceutical
composition containing the described substances, and a use thereof.
##STR00001##
Inventors: |
Jiang; Yueheng; (Shanghai,
CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
INVENTISBIO SHANGHAI LTD. |
Shanghai |
|
CN |
|
|
Family ID: |
55303876 |
Appl. No.: |
15/723785 |
Filed: |
October 3, 2017 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
15501387 |
Feb 2, 2017 |
9809580 |
|
|
PCT/CN2015/086605 |
Aug 11, 2015 |
|
|
|
15723785 |
|
|
|
|
Current U.S.
Class: |
514/307 |
Current CPC
Class: |
C07B 2200/05 20130101;
C07C 323/63 20130101; C07D 405/10 20130101; A61P 17/06 20180101;
C07C 319/20 20130101; A61P 9/10 20180101; A61P 9/12 20180101; A61K
31/4725 20130101; A61P 19/02 20180101; A61P 19/06 20180101; C07D
213/57 20130101; A61K 31/277 20130101; C07C 2601/02 20170501; A61P
43/00 20180101; A61P 5/20 20180101; A61P 9/00 20180101; A61P 35/00
20180101; C07D 213/68 20130101; C07D 213/70 20130101; C07D 213/74
20130101; A61K 31/4418 20130101; A61P 5/18 20180101; A61P 39/02
20180101; A61P 13/12 20180101; C07B 59/002 20130101; A61P 13/04
20180101; C07C 323/62 20130101; C07D 401/04 20130101 |
International
Class: |
C07D 405/10 20060101
C07D405/10; C07D 401/04 20060101 C07D401/04; C07D 213/57 20060101
C07D213/57; C07D 213/74 20060101 C07D213/74; C07D 213/70 20060101
C07D213/70; C07B 59/00 20060101 C07B059/00; C07C 323/63 20060101
C07C323/63; C07D 213/68 20060101 C07D213/68 |
Foreign Application Data
Date |
Code |
Application Number |
Aug 13, 2014 |
CN |
201410398333.4 |
Claims
1-16. (canceled)
17. A compound selected from the group consisting of ##STR00078##
##STR00079## or a pharmaceutically acceptable salt thereof.
18. The compound of claim 17, which is ##STR00080## or a
pharmaceutically acceptable salt thereof.
19. The compound of claim 17, which is ##STR00081## or a
pharmaceutically acceptable salt thereof.
20. The compound of claim 17, which is ##STR00082## or a
pharmaceutically acceptable salt thereof.
21. The compound of claim 17, which is ##STR00083## or a
pharmaceutically acceptable salt thereof.
22. The compound of claim 17, which is ##STR00084## or a
pharmaceutically acceptable salt thereof.
23. The compound of claim 17, which is ##STR00085## or a
pharmaceutically acceptable salt thereof.
24. The compound of claim 17, which is ##STR00086## or a
pharmaceutically acceptable salt thereof.
25. The compound of claim 17, which is ##STR00087## or a
pharmaceutically acceptable salt thereof.
26. The compound of claim 17, which is ##STR00088## or a
pharmaceutically acceptable salt thereof.
27. A pharmaceutical composition comprising the carboxylic acid
compound according to claim 17, or a pharmaceutically acceptable
salt thereof, and a pharmaceutically acceptable carrier.
28. A method of treating gout in a human, comprising administering
to the human an effective amount of a compound selected from the
group consisting of ##STR00089## ##STR00090## or a pharmaceutically
acceptable salt thereof.
29. The method of claim 28, comprising administering to the human
an effective amount of compound ##STR00091## or a pharmaceutically
acceptable salt thereof.
30. The method of claim 28, comprising administering to the human
an effective amount of compound ##STR00092## or a pharmaceutically
acceptable salt thereof.
31. The method of claim 28, comprising administering to the human
an effective amount of compound ##STR00093## or a pharmaceutically
acceptable salt thereof.
32. The method of claim 28, comprising administering to the human
an effective amount of compound ##STR00094## or a pharmaceutically
acceptable salt thereof.
33. The method of claim 28, comprising administering to the human
an effective amount of compound ##STR00095## or a pharmaceutically
acceptable salt thereof.
34. The method of claim 28, comprising administering to the human
an effective amount of compound ##STR00096## or a pharmaceutically
acceptable salt thereof.
35. The method of claim 28, comprising administering to the human
an effective amount of compound ##STR00097## or a pharmaceutically
acceptable salt thereof.
36. The method of claim 28, comprising administering to the human
an effective amount of compound ##STR00098## or a pharmaceutically
acceptable salt thereof.
37. The method of claim 28, comprising administering to the human
an effective amount of compound ##STR00099## or a pharmaceutically
acceptable salt thereof.
38. A method of reducing the blood uric acid level in a human,
comprising administering to the human an effective amount of a
compound selected from the group consisting of ##STR00100##
##STR00101## or a pharmaceutically acceptable salt thereof.
39. A method of treating hyperuricemia in a human, comprising
administering to the human an effective amount of a compound
selected from the group consisting of ##STR00102## ##STR00103## or
a pharmaceutically acceptable salt thereof.
Description
TECHNICAL FIELD
[0001] The invention relates to pharmaceutical technical field,
specifically, to carboxylic acid compounds and pharmaceutically
acceptable salts, prodrugs, and solvates thereof and preparation
methods thereof, and to pharmaceutical compositions comprising the
same and uses thereof.
BACKGROUND TECHNIQUE
[0002] Uric acid is the final metabolite of diet and purine in
human body. In vivo environment (pH 7.4, 37 degrees), uric acid is
present in blood mainly in the form of sodium salt of uric acid,
the serum uric acid value of normal people is generally lower than
6 mg/dL. When uric acid in serum exceeds 7 mg/dL (Shi, et al.,
Nature 2003, 425: 516-523), sodium salt of uric acid will
crystallize out and precipitate on joints and other parts of the
body, and result in disorders such as gout, urinary stones, kidney
stones, etc. Patients with gout are often accompanied with other
complications, including hypertension, diabetes, hyperlipidemia,
dyslipidemia, atherosclerosis, obesity, metabolic disease,
nephropathy, cardiovascular disease, and respiratory disease, etc.
(Rock, Et al., Nature Reviews Rheumatology 2013, 9: 13-23). In
2002, Japanese scientists Endou group reported that anion transport
channel protein URAT1 is a major protein responsible for
reabsorption of uric acid in kidney, they also found that the blood
uric acid in people with URAT1 gene mutation (causing the synthesis
of such protein being interrupted, inducing nonfunctional proteins)
is only one-tenth of that in normal people (Enomoto et. al., Nature
2002 417: 447-452). These findings in human genetics demonstrate
that URAT1 anion transport protein in kidney plays very important
role in concentration of uric acid in blood, and indicates that
URAT1 is a very good and specific target of a drug for reducing
blood uric acid.
[0003] The main objective in the treatment of gout and its
complications caused by higher level of blood uric acid is to
reduce blood uric acid to lower than 6 mg/dL, the main methods are
as follows: 1) to inhibit the generation of uric acid, such as
allopurinol, febuxostat, which are drugs for inhibiting Xanthine
oxidase; 2) to inhibit the reabsorption of uric acid, such as
benzbromarone and probenecid, and lesinurad which is currently in
clinical research, all of which are drugs for inhibiting kidney
URAT1 anion transport channel protein.
[0004] In addition to URAT1, there are other cation transport
channel proteins in kidney, such as Glut9 and OAT1 etc., which are
also found to be able to reabsorb uric acid back to blood from
renal tubules. Kidney is a major excretion pathway of uric acid in
human body (70%), intestinal system (via ABCG2 etc.,) is
responsible for excreting approximate 30% of uric acid (Sakurai,
et. al., Current Opinion in Nephology and Hypertension 2013, 22:
545-550).
[0005] Human urate anion transporter 1, hURAT1, a member of anion
transporter family, is located at luminal surface side of
epithelial cells of renal proximal convoluted tubules, mainly
participates in the reabsorption of uric acid in renal proximal
convoluted tubules. URAT1 accomplishes reabsorption of uric acid
and excretion of small amount of uric acid by exchanging univalent
anions within cells with uric acid in lumens. Anion transport
channel proteins located in renal proximal convoluted tubules also
comprise anion transport channel protein OAT4, which has 42% of
similarity with URAT1 (amino acids of protein). Therefore,
generally, a potent URAT1 inhibitor will also inhibit OAT4 and some
other anion transport channel proteins.
[0006] At present, all the clinical drugs for reducing blood uric
acid have some side effects, for example, allopurinol will cause
life-threatening hypersensitivity in some populations, febuxostat
has cardiovascular side effects, and benzbromarone has liver
toxicity and has been taken back by Sanofi from some markets.
Therefore, it is urgent to search for novel, efficient and
low-toxic drugs for reducing blood uric acid, and this will have
great clinical significance and application prospects.
[0007] Thioacetate compounds have been reported in the prior art,
e.g., a class of phenylthioacetate compounds were reported in
CN102939279A, a class of thioacetate compounds were reported in
CN103068801A, wherein thioacetate compounds in CN103068801A are
obtained from the compounds in CN102939279A by essentially
replacing carbons of benzene groups in skeletons of the compounds
in CN102939279A with 1 to 4 N atoms.
[0008] Since categories of drugs for gout treatment are very
limited in market, it is important to develop anti-gout drugs with
high efficiency and low toxicity.
CONTENTS OF THE INVENTION
[0009] According to one aspect of the present invention, one
objective of the present invention is to design and synthesize a
carboxylic acid compound, and pharmaceutically acceptable salts,
prodrugs, and solvates thereof.
[0010] According to another aspect of the present invention,
another objective of the present invention is to provide a
preparation method of the carboxylic acid compound, and
pharmaceutically acceptable salts, prodrugs, and solvates
thereof.
[0011] According to another aspect of the present invention,
another objective of the present invention is to provide a use of
the carboxylic acid compound, and pharmaceutically acceptable
salts, prodrugs, and solvates thereof in the preparation of a drug
for promoting the excretion of uric acid with URAT1 as a
target.
[0012] According to another aspect of the present invention,
another objective of the present invention is to provide a
pharmaceutical composition comprising one or more selected from the
carboxylic acid compound, and pharmaceutically acceptable salts,
prodrugs, and solvates thereof.
[0013] The carboxylic acid compound according to the present
invention is represented by the following Chemical Formula I:
##STR00002##
[0014] Wherein,
X is C or N;
[0015] Y, W and Z are each independently C or N; A is S, N,
SO.sub.2, O or absent; Q is substituted or unsubstituted C1-6
straight-chain or branched-chain alkylene, substituted or
unsubstituted C3-6 cycloalkylene, substituted or unsubstituted
C6-12 arylene, wherein substituent is --CD.sub.3, C1-6 alkyl, C3-6
cycloalkyl, C3-6 cycloalkylene or halogen; M is H, Na, K, Ca or
C1-4 alkyl; R.sup.1, R.sup.2 and R.sup.3 are each independently
hydrogen, halogen or absent; R.sup.a and R.sup.b are each
independently hydrogen, C1-6 alkyl or bond to each other to form a
substituted or unsubstituted C6-10 aromatic ring structure, wherein
the substituent in the substituted C6-10 aromatic ring structure is
halogen, C1-3 alkyl or C1-3 alkoxy; R.sup.c is --CN, carboxyl,
hydroxyl-substituted or unsubstituted C1-6 alkyl,
hydroxyl-substituted or unsubstituted C3-6 cycloalkyl,
hydroxyl-substituted or unsubstituted 3- to 6-membered
heterocycloalkyl containing 1 to 3 heteroatoms selected from O, S
and N.
[0016] Preferably,
X is C or N;
[0017] Y, W and Z are each independently C or N; A is S, N,
SO.sub.2, O or absent; Q is substituted or unsubstituted C1-3
straight-chain or branched-chain alkylene, substituted or
unsubstituted C3-5 cycloalkylene, phenyl, wherein substituent is
--CD.sub.3, C1-3 alkyl, C3-5 cycloalkyl, C3-5 cycloalkylene or
halogen selected from fluorine, chlorine, bromine and iodine; M is
H, Na, K, Ca or C1-4 alkyl; R.sup.1, R.sup.2 and R.sup.3 are each
independently hydrogen, halogen or absent; R.sup.a and R.sup.b are
each independently hydrogen, C1-3 alkyl or bond to each other to
form a substituted or unsubstituted benzene ring structure, wherein
the substituent in the substituted benzene ring structure is
halogen, C1-3 alkyl or C1-3 alkoxy; R.sup.c is --CN, carboxyl,
hydroxyl-substituted or unsubstituted C1-3 alkyl,
hydroxyl-substituted or unsubstituted C3-5 cycloalkyl,
hydroxyl-substituted or unsubstituted 3- to 5-membered
heterocycloalkyl containing 1 to 3 heteroatoms selected from O, S
and N.
[0018] More preferably,
X is C or N;
[0019] Y, W and Z are each independently C or N; A is S, N,
SO.sub.2, O or absent; Q is substituted or unsubstituted C1-3
straight-chain or branched-chain alkylene,
##STR00003##
(cis or trans),
##STR00004##
(cis or trans),
##STR00005##
phenyl, wherein substituent is methyl, ethyl, propyl, --CD.sub.3,
C3-5 cycloalkyl, C3-5 cycloalkylene or fluorine;
M is H;
[0020] R.sup.1, R.sup.2 and R.sup.3 are each independently
hydrogen, halogen or absent; R.sup.a and R.sup.b are each
independently hydrogen, or bond to each other to form a benzene
ring; R.sup.c is --CN, carboxyl, methyl, ethyl, propyl,
hydroxymethyl, hydroxyethyl, hydroxypropyl, cyclopropyl,
cyclobutyl, hydroxyl-substituted cyclopropyl, hydroxyl-substituted
cyclobutyl, oxiranyl, oxetanyl, hydroxyl-substituted oxiranyl or
hydroxyl-substituted oxetanyl.
[0021] Further preferably,
X is C or N;
[0022] Y, W and Z are each independently C or N;
A is S;
[0023] Q is substituted or unsubstituted ethylene, propylene,
isopropylidene,
##STR00006##
(cis or trans),
##STR00007##
(cis or trans),
##STR00008##
phenyl, wherein substituent is methyl, ethyl, propyl, --CD.sub.3,
cyclopropyl, cyclobutyl, cyclopentyl, cyclopropylidene,
cyclobutylidene, cyclopentylidene or fluorine;
M is H;
[0024] R.sup.1, R.sup.2 and R.sup.3 are each independently
hydrogen, halogen or absent; R.sup.a and R.sup.b are each
independently hydrogen, or bond to each other to form a benzene
ring; R.sup.c is --CN, carboxyl, methyl, ethyl, propyl,
hydroxymethyl, hydroxyethyl, hydroxypropyl, cyclopropyl,
cyclobutyl, hydroxyl-substituted cyclopropyl, hydroxyl-substituted
cyclobutyl, oxiranyl, oxetanyl, hydroxyl-substituted oxiranyl or
hydroxyl-substituted oxetanyl.
[0025] In one embodiment, the carboxylic acid compound according to
the present invention is represented by the following chemical
formula II:
##STR00009##
[0026] Wherein,
X is C or N;
[0027] Y, W and Z are each independently C or N; A is S, N,
SO.sub.2, O or absent; Q is substituted or unsubstituted C1-6
straight-chain or branched-chain alkylene, substituted or
unsubstituted C3-6 cycloalkylene, substituted or unsubstituted
C6-12 arylene, wherein substituent is --CD.sub.3, C1-6 alkyl, C3-6
cycloalkyl, C3-6 cycloalkylene or halogen; M is H, Na, K, Ca or
C1-4 alkyl; R.sup.1, R.sup.2 and R.sup.3 are each independently
hydrogen, halogen or absent; R.sup.a and R.sup.b are each
independently hydrogen, C1-6 alkyl or bond to each other to form a
substituted or unsubstituted C6-10 aromatic ring structure, wherein
the substituent in the substituted C6-10 aromatic ring structure is
halogen, C1-3 alkyl or C1-3 alkoxy; R.sup.c is --CN, carboxyl,
hydroxyl-substituted or unsubstituted C1-6 alkyl,
hydroxyl-substituted or unsubstituted C3-6 cycloalkyl,
hydroxyl-substituted or unsubstituted 3- to 6-membered
heterocycloalkyl containing 1 to 3 heteroatoms selected from O, S
and N.
[0028] Preferably,
X is C or N;
[0029] Y, W and Z are each independently C or N; A is S, N,
SO.sub.2, O or absent; Q is substituted or unsubstituted C1-3
straight-chain or branched-chain alkylene, substituted or
unsubstituted C3-5 cycloalkylene, phenyl, wherein substituent is
--CD.sub.3, C1-3 alkyl, C3-5 cycloalkyl, C3-5 cycloalkylene or
halogen selected from fluorine, chlorine, bromine and iodine; M is
H, Na, K, Ca or C1-4 alkyl; R.sup.1, R.sup.2 and R.sup.3 are each
independently hydrogen, halogen or absent; R.sup.a and R.sup.b are
each independently hydrogen, C1-3 alkyl or bond to each other to
form a substituted or unsubstituted benzene ring structure, wherein
the substituent in the substituted benzene ring structure is
halogen, C1-3 alkyl or C1-3 alkoxy; R.sup.c is --CN, carboxyl,
hydroxyl-substituted or unsubstituted C1-3 alkyl,
hydroxyl-substituted or unsubstituted C3-5 cycloalkyl,
hydroxyl-substituted or unsubstituted 3- to 5-membered
heterocycloalkyl containing 1 to 3 heteroatoms selected from O, S
and N.
[0030] More preferably,
X is C or N;
[0031] Y, W and Z are each independently C or N; A is S, N,
SO.sub.2, O or absent; Q is substituted or unsubstituted C1-3
straight-chain or branched-chain alkylene,
##STR00010##
(cis or trans),
##STR00011##
(cis or trans),
##STR00012##
phenyl, wherein substituent is methyl, ethyl, propyl, --CD.sub.3,
C3-5 cycloalkyl, C3-5 cycloalkylene or fluorine;
M is H;
[0032] R.sup.1, R.sup.2 and R.sup.3 are each independently
hydrogen, halogen or absent; R.sup.a and R.sup.b are each
independently hydrogen, or bond to each other to form a benzene
ring; R.sup.c is --CN, carboxyl, methyl, ethyl, propyl,
hydroxymethyl, hydroxyethyl, hydroxypropyl, cyclopropyl,
cyclobutyl, hydroxyl-substituted cyclopropyl, hydroxyl-substituted
cyclobutyl, oxiranyl, oxetanyl, hydroxyl-substituted oxiranyl or
hydroxyl-substituted oxetanyl.
[0033] Further preferably,
X is C or N;
[0034] Y, W and Z are each independently C or N;
A is S;
[0035] Q is substituted or unsubstituted ethylene, propylene,
isopropylidene,
##STR00013##
(cis or trans),
##STR00014##
(cis or trans),
##STR00015##
phenyl, wherein substituent is methyl, ethyl, propyl, --CD.sub.3,
cyclopropyl, cyclobutyl, cyclopentyl, cyclopropylidene,
cyclobutylidene, cyclopentylidene or fluorine;
M is H;
[0036] R.sup.1, R.sup.2 and R.sup.3 are each independently
hydrogen, halogen or absent; R.sup.a and R.sup.b are each
independently hydrogen, or bond to each other to form a benzene
ring; R.sup.c is --CN, carboxyl, methyl, ethyl, propyl,
hydroxymethyl, hydroxyethyl, hydroxypropyl, cyclopropyl,
cyclobutyl, hydroxyl-substituted cyclopropyl, hydroxyl-substituted
cyclobutyl, oxiranyl, oxetanyl, hydroxyl-substituted oxiranyl or
hydroxyl-substituted oxetanyl.
[0037] According to another embodiment of the present invention,
the carboxylic acid compound according to the present invention is
selected from the following specific compounds 1 to 41:
##STR00016## ##STR00017## ##STR00018## ##STR00019## ##STR00020##
##STR00021## ##STR00022## ##STR00023##
[0038] In the present invention, the pharmaceutically acceptable
salts of the compounds in the present invention are not
particularly limited, as long as they are pharmaceutically
acceptable, examples include, but are not limited to, ammonium
salts, alkali metal salts and alkaline-earth metal salts, such as
ammonium salts, sodium salts, potassium salts, calcium salts and
the like.
[0039] The present invention also includes isotopically-labeled
compounds of the present invention, these isotopically-labeled
compounds are identical to those recited herein, except that one or
more atoms have an atomic mass or mass number different from the
atomic mass or mass number usually found in nature. Examples of
isotopes that can be incorporated into compounds of the invention
include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine
and chloride, such as .sup.2H, .sup.3H, .sup.13C, .sup.14C,
.sup.15N, .sup.18O, .sup.17O, .sup.18F and .sup.36Cl.
[0040] Certain isotopically-labeled compounds of the present
invention (for example, compounds labeled with .sup.3H and
.sup.14C) can be used in compounds and/or substrate tissue
distribution assays. Tritiated (i.e., .sup.3H) and carbon-14 (i.e.,
.sup.14C) isotopes are particularly preferred for their ease of
preparation and detection. Further, substitution with heavy
isotopes such as deuterium (i.e., .sup.2H) may produce certain
therapeutic advantages resulting from greater metabolic stability
(for example, extension of half-life in vivo or reduction of dose
requirements), and thus being preferably used in certain
conditions. The isotopically-labeled compounds of the present
invention may generally be prepared by replacing non-isotope
labeled agents with isotopically-labeled agents, by following the
procedures similar to those disclosed in the flow routes and/or
examples below.
[0041] In the present invention, the prodrugs of the compounds of
the present invention are not particularly limited, as long as they
can be metabolized in vivo into the compounds of the present
invention, examples include, but are not limited to, esters etc.,
such as methyl ester, ethyl ester and the like.
[0042] Further objective of the present invention is to provide a
preparation process of the carboxylic acid compound, and
pharmaceutically acceptable salts, prodrugs, and solvates thereof,
the process comprises:
Reaction Route 1:
##STR00024##
[0044] Step 1: The starting reactant 1-1 is dissolved in dioxane,
then potassium acetate, bis(pinacolato)diboron (B.sub.2(pin).sub.2)
and palladium catalyst
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium are added
thereto, the mixture is heated and reacted until the reaction is
completed. The reaction solution is cooled, quenched by addition of
ice water, extracted with ethyl acetate, then the organic phase is
combined and washed with saturated brine. The organic phase is
dried over sodium sulfate, then rotatory evaporated and purified by
column chromatography, to give compound (1-2).
[0045] Step 2: 3-bromo-4-chloropyridine or 2-bromo-1-chlorobenzene
is dissolved in dimethyl formamide and water, compound (1-2)
obtained in step 1, sodium carbonate, palladium catalyst
[1,1'-bis(diphenylphosphino) ferrocene]dichloropalladium are added
thereto, then the resulting mixture is heated and reacted. The
reaction solution is cooled, quenched in ice water, extracted with
ethyl acetate, washed with saturated brine, and dried over
anhydrous sodium sulfate, then rotatory evaporated and purified by
column chromatography, to give compound (1-3).
[0046] Step 3: To compound (1-3) obtained in step 2 dissolved in
dimethyl formamide, is added sodium sulfide, the resulting mixture
is heated and reacted, then cooled down to room temperature, after
that, anhydrous potassium carbonate and a reactant
##STR00025##
(which is determined by the structure of the final products) are
added thereto, the reaction is carried out at higher temperature
until the reaction is completed. The reaction solution is cooled,
quenched in ice water and extracted with ethyl acetate, then the
organic phase is dried over anhydrous sodium sulfate, filtered and
rotatory evaporated, to give compound (1-4). The crude product is
directly subject to the next step.
[0047] Step 4: The reaction is carried out overnight between
compound (1-4) obtained in step 3 and lithium hydroxide in
tetrahydrofuran and water at room temperature. Tetrahydrofuran is
removed by concentration, and aqueous phase is extracted with
dichloromethane and collected. Aqueous phase is then adjusted to pH
4-5 using 2N of hydrochloric acid regulating system, and extracted
with dichloromethane. The organic phase is combined, dried and
rotatory evaporated, to give the final compound of chemical formula
(1-5).
Reaction Route 2:
##STR00026##
[0049] The reaction is carried out overnight between compound (1-3)
and a reactant
##STR00027##
(which is determined by the structure of the final product) in
phenol at higher temperature. Then, the reaction solution is cooled
down to room temperature, added with ether and filtered. The filter
cake is purified by preparative reverse phase chromatography to
give the final product (2-1).
Reaction Route 3:
##STR00028##
[0051] Step 1: A reactant
##STR00029##
which is determined by the structure of final product,
triphenylphosphine and diethyl azodicarboxylate are sequentially
added to 3-bromopyridin-4-ol or 2-bromophenol (3-1) dissolved in
tetrahydrofuran under the protection of nitrogen at 0.degree. C.,
then the mixture is warmed to room temperature and reacted. The
reaction solution is directly concentrated and then purified by
preparative silica gel plate to give compound (3-2).
[0052] Step 2: The resulting compound (3-2), aqueous solution of
sodium carbonate, compound (1-2) and tetrakis(triphenylphosphine)
palladium(0) are added to dioxane, heated to 80.degree. C. and
reacted for 12 hours. Then the reaction solution is cooled down to
room temperature, the reaction solution is added with ethyl
acetate, and washed with water and brine. The organic phase is
dried, filtered, concentrated and purified by preparative silica
gel plate, to give compound (3-3).
[0053] Step 3: Compound (3-3), lithium hydroxide or sodium
hydroxide are added to tetrahydrofuran/water, and the mixture was
reacted at room temperature for hours. Then the pH of the reaction
solution is adjusted with concentrated hydrochloric acid, and the
reaction solution is added with ethyl acetate, washed with water
and brine. The organic phase is dried, filtered, concentrated and
purified by preparative silica gel plate, to give compound
(3-4).
Reaction Route 4:
##STR00030##
[0055] Step 1: The starting reactant 4-1 is dissolved in dioxane,
then potassium acetate, bis(pinacolato)diboron (B.sub.2(pin).sub.2)
and palladium catalyst
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium are added
thereto, then the mixture is heated and reacted until the reaction
is completed. The reaction solution is cooled, quenched by addition
of ice water and extracted with ethyl acetate, then the organic
phase is combined and washed with saturated brine. The organic
phase is dried over sodium sulfate, then rotatory evaporated and
purified by column chromatography, to give compound (4-2).
[0056] Step 2: 3-bromo-4-chloropyridine or 2-bromo-1-chlorobenzene
is dissolved in dimethyl formamide and water, compound (4-2)
obtained in step 1, sodium carbonate, palladium catalyst
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium are added
thereto at the same time, then the resulting mixture is heated and
reacted. The reaction solution is cooled, quenched in ice water,
extracted with ethyl acetate, washed with saturated brine, dried
over sodium sulfate, then rotatory evaporated and purified by
column chromatography, to give compound (4-3).
[0057] Step 3: To compound (4-3) obtained in step 2 dissolved in
dimethyl formamide, is added sodium sulfide, the resulting mixture
is heated and reacted, then cooled down to room temperature, after
that, anhydrous potassium carbonate and a reactant
##STR00031##
(which is determined by the structure of the final product) are
added thereto, and the reaction is carried out at higher
temperature until the reaction is completed. The reaction solution
is cooled, quenched in ice water and extracted with ethyl acetate,
then the organic phase is dried over sodium sulfate, filtered and
rotatory evaporated, to give compound (4-4). The crude product is
directly subject to the next step.
[0058] Step 4: The reaction is carried out overnight between
compound (4-4) obtained in step 3 and lithium hydroxide in
tetrahydrofuran and water at room temperature. Tetrahydrofuran is
removed by concentration, and aqueous phase is extracted with
dichloromethane and collected. Then the aqueous phase is adjusted
to pH 4-5 using 2N of hydrochloric acid regulating system, and
extracted with dichloromethane. The organic phase is combined,
dried, then rotatory evaporated, to give the final compound of
chemical formula (4-5).
Reaction Route 5:
##STR00032##
[0060] Step 1: 3-bromo-4-methylpyridine is dissolved in
tetrahydrofuran, the solution is cooled and then lithium
diisopropylamide (LDA) is added thereto and reacted, then the
reactant
##STR00033##
which is determined by the structure of the final product, is added
dropwise thereto and the reaction is continued. Thereafter, the
reaction is quenched with saturated sodium bicarbonate solution,
and then the reaction solution is added with ethyl acetate, washed
with water and brine. The organic phase is dried, filtered,
concentrated and purified by preparative silica gel plate, to give
compound (5-1).
[0061] Step 2: Compound (5-1), aqueous solution of sodium
carbonate, compound (1-2),
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium are added
to dimethyl formamide, heated until they reacted. The reaction
solution is added with ethyl acetate, washed with water and brine.
The organic phase is dried, filtered, concentrated and purified by
preparative silica gel plate, to give compound (5-2).
[0062] Step 3: Compound (5-2) and lithium hydroxide are added to
tetrahydrofuran/water and reacted at room temperature. The pH of
reaction solution is adjusted with dilute hydrochloric acid (1 M),
and then the reaction solution is added with ethyl acetate, washed
with water and brine. The organic phase is dried, filtered,
concentrated and purified by preparative silica gel plate, to give
compound (5-3).
Reaction Route 6:
##STR00034##
[0064] Step 1: Compound (1-3), anhydrous potassium carbonate and
reactant
##STR00035##
(or reactant
##STR00036##
are added to dimethyl formamide, stirred at room temperature, then
stirred at higher temperature. The mixture is then cooled to room
temperature, added with water and ethyl acetate, after that, the
organic layer is washed with saturated brine, dried over sodium
sulfate, and the solvent is distilled off under reduced pressure,
to obtain a crude brown oil which is purified by column
chromatography to yield compound (6-1-1) and compound (6-2-1)
respectively.
[0065] Step 2: Compound (6-1-1) and compound (6-2-1) are dissolved
in tetrahydrofuran respectively, and are slowly added by dropwise
to suspension of sodium hydride in dimethyl formamide at 0.degree.
C. while stirring, a solution of Iodomethane-d3 in dimethyl
formamide is further added thereto at 0.degree. C., then the
resulting mixture is stirred overnight at room temperature. The
reaction is quenched by addition water, its pH is adjusted with 1 N
hydrochloric acid, then the solvent is distilled off under reduced
pressure, and the remaining oily substance is purified by
preparative HPLC, to give compound (6-1-2) and compound (6-2-2)
respectively.
[0066] Step 3: Compound (6-1-2) and lithium hydroxide are added to
tetrahydrofuran/water (3 mL/1 mL), and reacted at room temperature.
The reaction solution is adjusted to pH 4 with dilute hydrochloric
acid (1 M), then the reaction solution is added with ethyl acetate
and washed with brine. The organic phase is dried, filtered,
concentrated and preparatively purified, to give compound
(6-1-3).
[0067] According to another aspect of the invention, further
provided herein is a use of the carboxylic acid compound, and
pharmaceutically acceptable salts, prodrugs, and solvates thereof
in the preparation of a drug that promotes the excretion of uric
acid, preferably, a drug that promotes the excretion of uric acid
with URAT1 as a target.
[0068] According to another aspect of the invention, further
provided herein is a pharmaceutical composition comprising one or
more selected from the carboxylic acid compound, and
pharmaceutically acceptable salts, prodrugs, and solvates thereof,
and optionally a pharmaceutically acceptable carrier.
[0069] According to another aspect of the invention, further
provided herein is a use of the carboxylic acid compound, and
pharmaceutically acceptable salts, prodrugs, and solvates thereof
or pharmaceutical composition thereof in the preparation of a drug
for the treatment or prevention of a disease or disorder caused by
abnormal organ or tissue levels of uric acid in an individual.
[0070] Wherein, the disease or disorder caused by abnormal organ or
tissue levels of uric acid in an individual includes: gout, gouty
arthritis, a recurrent gout attack, hyperuricemia, joint
inflammation, arthritis, urolithiasis, kidney disease, kidney
stones, kidney failure, hypertension, cardiovascular disease,
coronary heart disease, Lesch-Nyhan syndrome, Kelley-Seegmiller
syndrome, plumbism, hyperparathyroidism, psoriasis and
sarcoidosis.
[0071] Preferably, the disease or disorder is hyperuricemia in
human and animals, or gout in human and animals.
[0072] According to another aspect of the invention, further
provided herein is a use of the carboxylic acid compound, and
pharmaceutically acceptable salts, prodrugs, and solvates thereof
or the pharmaceutical composition thereof in the preparation of a
drug for lowering blood levels of uric acid in human and
animals.
[0073] According to another aspect of the invention, a use of the
carboxylic acid compound, and pharmaceutically acceptable salts,
prodrugs, and solvates thereof or the pharmaceutical composition
thereof in the preparation of a drug for lowering blood levels of
uric acid in human and animals.
[0074] Further provided herein is a combination of the carboxylic
acid compound, and pharmaceutically acceptable salts, prodrugs, and
solvates thereof or a pharmaceutical composition thereof with a
second agent effective for the treatment of gout.
[0075] Wherein, the second agent is a xanthine oxidase inhibitor, a
xanthine dehydrogenase inhibitor, a xanthine oxidoreductase
inhibitor, or a combination thereof, preferably, allopurinol,
febuxostat or a combination thereof.
[0076] The pharmaceutical composition or drugs provided herein can
be in various forms, such as tablet, capsule, powder, syrup,
solution, suspension and aerosol, etc., and can be present in
suitable solid or liquid carriers or diluents and in disinfected
instruments suitable for injection or infusion.
[0077] Various formulations of the pharmaceutical compositions or
drugs of the present invention can be prepared according to
conventional preparation processes in the pharmaceutical field.
Unit dosage of these formulations contains 0.05 mg-200 mg compounds
of formula (I) or (II), preferably, unit dosage of these
formulations contains 0.1 mg-100 mg compounds of formula (I) or
(II).
[0078] The compounds and pharmaceutical compositions of the present
invention can be clinically used for mammals, including human and
animals, and can be administered via oral, nasal, dermal,
pulmonary, or gastrointestinal etc. administration routes, Oral
route is the most preferable. The optimal preferred daily dosage is
0.001-10 mg/kg body weight, administered at one time, or 0.001-10
mg/kg body weight administered at divided doses. No matter what
kinds of administration manners, the optimal dosage for an
individual should be determined by specific treatment. Generally,
the most suitable dosage can be obtained by gradually increasing
dosages from a smaller dosage.
[0079] In the present invention, term "effective amount" may refer
to an effective amount for dose and period of time required to
achieve predicted effects. The effective amount may vary due to
some factors, such as categories of diseases or syndromes of
diseases during treatment, construction of a specific targeted
organ being administered, body size of a patient or severity of
diseases or syndromes. Without excessive experiments, those with
common knowledge in the art can determine an effective amount for a
specific compound by experience.
Technical Effects
[0080] The research on anti-uric acid activity demonstrates that
the compounds of the present invention have superior activities in
inhibiting the reabsorption of uric acid, and can be used as novel
drugs for efficiently decreasing blood uric acid level, especially
as URAT1 inhibitors.
SPECIFIC EMBODIMENTS
[0081] The present invention will be explained with reference to
the specific examples below. It should be understood that, these
examples are merely used for illustrating the present invention but
not for limiting the scope of the invention. The experimental
methods in the following examples, when detailed conditions are not
specified, are carried out according to conventional conditions, or
according to conditions provided or constructed by manufacturers.
Unless defined or illustrated otherwise, all professional and
scientific terms used herein have the same meaning as commonly
known by a person skilled in the art. Additionally, any methods and
materials similar or equivalent to the recorded contents can be
used in the methods of the present invention.
[0082] In the synthetic processes of the following examples,
starting materials were obtained from commercial sources, such as
from Alfa Aesar (China) Chem Co. Ltd., Accela ChemBio Co. Ltd,
PharmaBlock Sciences (Nanjing), Inc., Dalian Ally Chem Co. Ltd.,
Tianjin Fuchen chemical reagent factory, Beijing Jingqiu chemical
product Co. Ltd., Zhangjiagang Aimate Chem Co. Ltd, Sinopharm
chemical reagent Shaanxi Co., Ltd, etc.
EXAMPLES
Example 1: Synthesis of Compound 1
##STR00037##
[0083] Step 1: Synthesis of
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1-naphthonitrile
(1-a)
[0084] In a three-necked flask (100 mL), potassium acetate (3.9 g,
39.8 mmol), bis(pinacolato)diboron (B.sub.2(pin).sub.2) (4.0 g,
15.75 mmol) and palladium catalyst
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (0.54 g,
0.66 mmol) were added to 4-bromo-1-naphthonitrile (3.0 g, 15.75
mmol) solution in dioxane (40 mL) under the protection of N.sub.2,
the mixture was heated to 90.degree. C. and reacted for 3 hours
until the reaction was completed. The reaction solution was cooled,
quenched by addition of 100 mL ice water and extracted with ethyl
acetate (100 mL, 3 times), then the organic phase was combined and
washed with saturated brine (100 mL, 3 times). The organic phase
was dried over sodium sulfate, then rotatory evaporated and
purified by column chromatography (petroleum ether/ethyl
acetate=20:1-10:1), to give an off-white solid product
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1-naphthonitrile
(1-a).
Step 2: Synthesis of 4-(4-chloropyridin-3-yl)-1-naphthonitrile
(1-b)
[0085] In a three-necked flask (100 mL),
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1-naphthonitrile
(2.4 g, 8.6 mmol) obtained in step 1, sodium carbonate (2.8 g,
26.42 mmol), palladium catalyst
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (0.35 g,
0.43 mmol) were added at the same time to 3-bromo-4-chloropyridine
(1.6 g, 8.31 mmol) solution in dimethyl formamide (40 mL) and water
(4.8 mL) under the protection of N.sub.2, then the mixture was
heated to 130.degree. C. and reacted for 5 hours. The reaction
solution was cooled, quenched by addition of 100 mL ice water,
extracted with ethyl acetate (100 mL, 3 times), washed with
saturated brine (100 mL, 3 times) and dried over sodium sulfate,
then rotatory evaporated and purified by column chromatography
(petroleum ether/ethyl acetate=10:1.about.petroleum ether/ethyl
acetate/dichloromethane=1:1:1), to give an off-white solid product
4-(4-chloropyridin-3-yl)-1-naphthonitrile (1-b).
Step 3: Synthesis of methyl 1-(((3-(4-cyanonaphthalen-1-yl)
pyridin-4-yl) thio)methyl)cyclopropanecarboxylate (1-c)
[0086] In a three-necked flask (100 mL), under the protection of
N.sub.2, 4-(4-chloropyridin-3-yl)-1-naphthonitrile (200 mg, 0.70
mmol) obtained in step 2 was dissolved in dimethyl formamide (20
mL), sodium sulfide (355 mg, 4.50 mmol) was added thereto, then the
mixture was heated to 130.degree. C. and reacted for about 1 hour,
after being cooled down to room temperature, anhydrous potassium
carbonate (523 mg, 3.70 mmol) and methyl
1-(bromomethyl)cyclopropanecarboxylate (440 mg, 2.20 mmol) are
sequentially added thereto, and then the resulting mixture was
heated to 130.degree. C. and further reacted for about 1.1 hour
until the reaction was completed. The reaction solution was cooled,
added with 100 mL ice water to quench the reaction and extracted
with ethyl acetate (100 mL, 3 times), then the organic phase was
dried over anhydrous sodium sulfate, filtered and rotatory
evaporated, to give 450 mg of yellow oily product methyl
1-(((3-(4-cyanonaphthalen-1-yl) pyridin-4-yl)thio)methyl)
cyclopropanecarboxylate (1-c). The crude product was directly
subject to the next step.
Step 4: Synthesis of
1-(((3-(4-cyanonaphthalen-1-yl)pyridin-4-yl)thio)
methyl)cyclopropanecarboxylic acid (compound 1)
[0087] Methyl 1-(((3-(4-cyanonaphthalen-1-yl)
pyridin-4-yl)thio)methyl) cyclopropanecarboxylate (450 mg, 1.20
mmol) obtained in step 3, lithium hydroxide (90 mg, 3.70 mmol),
tetrahydrofuran (30 mL) and water (10 mL) were added into a
three-necked flask (100 mL) under the protection of N.sub.2, and
the mixture was reacted overnight at room temperature. The
resulting mixture was concentrated to remove tetrahydrofuran,
aqueous phase was extracted 3 times with dichloromethane (50 mL),
and the aqueous phase was collected. The aqueous phase was adjusted
to pH=4-5 with 2N of hydrochloric acid regulating system, and then
extracted with dichloromethane (100 mL, 3 times). The organic phase
was combined, dried with sodium sulfate, and then rotatory
evaporated. The crude product was prepared by high pressure to
yield compound 1, a white solid product.
[0088] LC-MS (ES, m/z): 361 [M+H].sup.+. H-NMR (300 MHz,
d.sub.6-DMSO, ppm): .delta. 0.86-0.94 (m, 2H), 1.08-1.14 (m, 2H),
3.38 (s, 2H), 7.54-7.57 (d, J=8.1 Hz, 1H), 7.65-7.91 (m, 4H),
8.23-8.26 (d, J=8.4 Hz, 1H), 8.31-8.33 (d, J=7.5 Hz, 1H), 8.49 (s,
1H), 8.69-8.71 (d, J=6 Hz, 1H).
Example 2: Synthesis of Compound 2
##STR00038##
[0090] Compound 2 was synthesized by a method similar to that in
Example 1, except that methyl
1-(bromomethyl)cyclopropanecarboxylate was replaced with the
corresponding compound in step 3.
[0091] LC-MS (ES, m/z): 335 [M+H].sup.+. H-NMR (300 MHz,
CD.sub.3OD, ppm): .delta.2.61-2.65 (m, 2H), 3.23-3.28 (m, 2H),
7.53-7.66 (m, 4H), 7.78-7.83 (m, 1H), 8.11-8.13 (d, J=7.2 Hz, 1H),
8.29-8.32 (d, J=8.4 Hz, 2H), 8.56 (m, 1H).
Example 3: Synthesis of Compound 3
##STR00039##
[0093] Compound 3 was synthesized by a method similar to that in
Example 1, except that methyl
1-(bromomethyl)cyclopropanecarboxylate was replaced with the
corresponding compound in step 3.
[0094] LC-MS (ES, m/z): 349 [M+H].sup.+. H-NMR (300 MHz,
CDCl.sub.3, ppm): .delta.1.69-1.79 (m, 2H), 2.24-2.29 (m, 2H),
2.95-3.00 (m, 2H), 7.47-7.50 (d, J=8.4 Hz, 1H), 7.57-7.70 (m, 3H),
7.82-7.88 (m, 1H), 8.12-8.31 (m, 3H), 8.58-8.60 (d, J=5.4 Hz, 1H),
12.15 (br, 1H).
Example 4: Synthesis of Compound 18
##STR00040##
[0095] Step 1: Synthesis of 2,2-dimethylthiirane (18-a)
[0096] Potassium thiocyanate (9.7 g, 0.1 mol) was dissolved in
water (10 mL), and 2,2-dimethyloxirane (7.2 g, 0.1 mol) was added
thereto at room temperature. After the mixture was reacted for 4
hours, the supernatant was added dropwise to the aqueous solution
(5 mL) of potassium thiocyanate (5 g, 0.05 mol), and the reaction
was lasted for another 16 hours. The reaction solution was added
with ether (50 mL) and water (30 mL), and the organic phase was
further washed with brine (10 mL). Then the organic phase was
dried, filtered and concentrated to yield a yellow oily
product.
Step 2: Synthesis of 4-mercapto-4-methylpentanenitrile (18-b)
[0097] In a three-necked flask (100 mL), n-butyllithium (2.5 M in
hexane, 3.6 mL, 9 mmol) was dissolved in tetrahydrofuran (10 mL)
under the protection of nitrogen, the solution was cooled to
-78.degree. C., then acetonitrile (378 mg, 9 mmol) was added
thereto, the mixture was reacted for 0.5 hour, to which was further
added dropwise a solution of 18-a (800 mg, 9 mmol) in
tetrahydrofuran (20 mL), the resulting mixture was heated to room
temperature and reacted for another 4 hours. The reaction was
quenched with 1 N hydrochloric acid (9 mL) at 0.degree. C., then
the reaction solution was added with ethyl acetate (50 mL), then
washed with brine (10 mL). The organic phase was dried, filtered
and concentrated to yield a yellow oily product.
Step 3: Synthesis of
4-(4-((4-cyano-2-methylbutan-2-yl)thio)pyridin-3-yl)-1-naphthonitrile
(18-c)
[0098] In a single-necked flask (100 mL), 18-b (600 mg, 4.65 mmol),
anhydrous potassium carbonate (641 mg, 4.65 mmol) and 1-b (300 mg,
1.13 mmol) were added to dimethyl formamide (15 mL), the mixture
was heated to 130.degree. C. and reacted for 2 hours. The reaction
solution was cooled, added with ethyl acetate (50 mL), washed with
water (30 mL) and brine (30 mL). The organic phase was dried,
filtered, concentrated, and purified by silica gel column
(petroleum ether/ethyl acetate=1:1) to yield a white solid.
Step 4: Synthesis of
4-((3-(4-cyanonaphthalen-1-yl)pyridin-4-yl)thio)-4-methylpentanoic
acid (18)
[0099] 18-c (190 mg, 0.53 mmol) and 1 M of aqueous solution of
sodium hydroxide (2.1 mL, 2.1 mmol,) were added to
tetrahydrofuran/methanol (2 mL/8 mL) in a single-necked flask (50
mL), and the mixture was reacted at 65.degree. C. for 36 hours. The
reaction solution was concentrated, adjusted to pH=4 with 1N
hydrochloric acid, then it was added with ethyl acetate (50 mL) and
washed with brine (50 mL). The organic phase was dried, filtered,
concentrated and purified by preparative reverse phase
chromatography to yield a white solid product.
[0100] LC-MS (ES, m/z): 377 [M+H].sup.+; H-NMR (400 MHz, DMSO-d6,
ppm): .delta. 12.00 (s, 1H), 8.62 (d, J=5.2 Hz, 1H), 8.44 (s, 1H),
8.18-8.23 (m, 2H), 7.81 (m, 1H), 7.72 (d, J=5.2 Hz, 1H), 7.65 (m,
1H), 7.55 (d, J=7.6 Hz, 1H), 7.39 (d, J=8.4 Hz, 1H), 2.02 (m, 2H),
1.72 (m, 2H), 1.13 (s, 6H).
Example 5: Synthesis of Compound 4
##STR00041##
[0101] Step 1: Synthesis of methyl
2-1-(((3-(4-cyanonaphthalen-1-yl) pyridin-4-yl)thio)methyl)
cyclopropyl)acetate (4-a)
[0102] In a three-necked flask (100 mL), under the protection of
N.sub.2, anhydrous potassium carbonate (523 mg, 3.70 mmol) and
methyl 2-(1-(mercaptomethyl) cyclopropyl)acetate (300 mg, 1.90
mmol) were sequentially added to
4-(4-chloropyridin-3-yl)-1-naphthonitrile (200 mg, 0.70 mmol)
solution in dimethyl formamide (20 mL), the mixture was heated to
130.degree. C. and reacted for about 2 hour until the reaction was
completed. The reaction solution was cooled, added with 100 mL ice
water to quench the reaction, and extracted with ethyl acetate (100
mL, 3 times), then organic phase was reversely washed with
saturated brine (100 mL, 4 times). The organic phase was dried over
anhydrous sodium sulfate, and then rotatory evaporated to yield a
yellow oily product. The crude product was directly subject to the
next step.
Step 2: Synthesis of
2-(1-(((3-(4-cyanonaphthalen-1-yl)pyridin-4-yl)
thio)methyl)cyclopropyl)acetic acid (compound 4)
[0103] Methyl 2-(1-(((3-(4-cyanonaphthalen-1-yl)
pyridin-4-yl)thio)methyl) cyclopropyl)acetate (417 mg, 1.07 mmol)
obtained in step 1, lithium hydroxide (78 mg, 3.26 mmol),
tetrahydrofuran (30 mL) and water (10 mL) were added into a
three-necked flask (100 mL) under the protection of N.sub.2, and
the mixture was reacted overnight at room temperature. Then, the
resulting solution was concentrated to remove tetrahydrofuran,
aqueous phase was extracted 3 times with dichloromethane (50 mL),
and the aqueous phase was collected. After that, the aqueous phase
was adjusted to pH=4-5 with 2N of hydrochloric acid regulating
system, and then extracted with dichloromethane (100 mL, 3 times).
The organic phase was combined, dried over sodium sulfate, rotatory
evaporated, prepared by high pressure, and then rotatory
evaporated. The resultant was lyophilized to yield a white
solid.
[0104] LC-MS (ES, m/z): 375 [M+H].sup.+. H-NMR (300 MHz,
CDCl.sub.3, ppm): .delta. 0.43 (m, 4H), 2.06-2.18 (m, 2H),
3.13-3.22 (m, 2H), 7.48-7.70 (m, 4H), 7.83-7.88 (m, 1H), 8.22-8.30
(m, 3H), 8.54-8.56 (d, J=5.4 Hz, 1H), 12.23 (br, 1H).
Example 6: Synthesis of Compound 5
##STR00042##
[0106] Compound 5 was synthesized by a method similar to that in
Example 5, except that methyl
2-(1-(mercaptomethyl)cyclopropyl)acetate was replaced with the
corresponding compound in step 1.
[0107] LC-MS (ES, m/z): 375 [M+H].sup.+. H-NMR (300 MHz,
CDCl.sub.3, ppm): .delta. 1.92-2.01 (m, 2H), 2.34-2.47 (m, 3H),
2.97-3.03 (m, 3H), 7.37-59 (m, 3H), 7.72-7.77 (t, J=7.8 Hz, 1H),
7.99-8.01 (d, J=7.2 Hz, 1H), 8.33-8.36 (d, J=8.4 Hz, 1H), 8.57-8.59
(d, J=5.4 Hz, 2H), 8.61-8.66 (m, 1H).
Example 7: Synthesis of Compound 6
##STR00043##
[0109] Compound 6 was synthesized by a method similar to that in
Example 5, except that methyl
2-(1-(mercaptomethyl)cyclopropyl)acetate was replaced with the
corresponding compound in step 1.
[0110] LC-MS (ES, m/z): 375 [M+H].sup.+. H-NMR (300 MHz,
CDCl.sub.3, ppm): .delta. 1.92-2.01 (m, 2H), 2.36-2.2.44 (m, 2H),
2.65-2.70 (m, 1H), 2.99-3.09 (m, 3H), 7.30-7.59 (m, 4H), 7.71-7.77
(m, 1H), 7.99-8.01 (d, J=7.2 Hz, 1H), 8.33-8.34 (m, 2H), 8.58-8.60
(m, 1H).
Example 8: Synthesis of Compound 12
##STR00044##
[0112] Compound 12 was synthesized by a method similar to that in
Example 5, except that methyl
2-(1-(mercaptomethyl)cyclopropyl)acetate was replaced with the
corresponding compound in step 1.
[0113] LC-MS (ES, m/z): 385 [M+H].sup.+. H-NMR (400 MHz,
d.sub.6-DMSO, ppm): .delta. 8.60 (d, J=6.4 Hz, 1H), 8.31 (s, 1H),
8.25 (d, J=7.6 Hz, 1H), 8.20 (d, J=8.4 Hz, 1H), 7.84-7.80 (m, 1H),
7.67-7.63 (m, 1H), 7.59 (d, J=7.2 Hz, 1H), 7.50 (d, J=6.4 Hz, 1H),
7.46 (d, J=8.4 Hz, 1H), 3.09 (t, J=8.0 Hz, 2H), 2.33-2.29 (m,
2H).
Example 9: Synthesis of Compound 13
##STR00045##
[0115] Compound 13 was synthesized by a method similar to that in
Example 5, except that methyl
2-(1-(mercaptomethyl)cyclopropyl)acetate was replaced with the
corresponding compound in step 1.
[0116] LC-MS (ES, m/z): 383 [M+H].sup.+. H-NMR (300 MHz,
d.sub.6-DMSO, ppm): .delta. 7.19-7.21 (d, J=5.7 Hz, 1H), 7.45-7.56
(m, 3H), 7.67-7.72 (m, 3H), 7.83-7.90 (m, 2H), 8.22-8.30 (m, 2H),
8.60-8.62 (d, J=6.3 Hz, 2H).
Example 10: Synthesis of Compound 14
##STR00046##
[0118] Compound 14 was synthesized by a method similar to that in
Example 5, except that methyl
2-(1-(mercaptomethyl)cyclopropyl)acetate was replaced with the
corresponding compound in step 1.
[0119] LC-MS (ES, m/z): 383 [M+H].sup.+. H-NMR (300 MHz,
d.sub.6-DMSO, ppm): .delta. 7.02-7.04 (d, J=5.7 Hz, 1H), 7.58-7.78
(m, 5H), 7.87-7.91 (m, 2H), 8.01-8.03 (d, J=7.8 Hz, 1H), 8.24-8.27
(d, J=8.4 Hz, 1H), 8.31-8.34 (d, J=7.5 Hz, 1H), 8.54-8.57 (m,
2H).
Example 11: Synthesis of Compound 15
##STR00047##
[0121] Compound 15 was synthesized by a method similar to that in
Example 5, except that methyl
2-(1-(mercaptomethyl)cyclopropyl)acetate was replaced with the
corresponding compound in step 1.
[0122] LC-MS (ES, m/z): 383 [M+H].sup.+. H-NMR (300 MHz,
CD.sub.30OD, ppm): .delta. 7.31-7.33 (d, J=6.3 Hz, 1H), 7.62-7.65
(d, J=8.1 Hz, 2H), 7.72-7.79 (m, 3H), 7.86-7.91 (m, 1H), 8.12-8.15
(d, J=8.4 Hz, 2H), 8.19-8.22 (d, J=7.5 Hz, 1H), 8.35-8.37 (d, J=8.4
Hz, 1H), 8.55-8.57 (d, J=6.3 Hz, 1H), 8.63 (s, 1H).
Example 12: Synthesis of Compound 20
##STR00048##
[0123] Step 1: Synthesis of 4-(4-chloropyridin-3-yl)benzonitrile
(20-b)
[0124] 3-bromo-4-chloropyridine (573 mg, 3 mmol), aqueous solution
of sodium carbonate (6 mL, 12 mmol, 2 M), 4-cyanophenylboronic acid
(441 mg, 3 mmol) and tetrakis(triphenylphosphine)palladium (0) (173
mg, 0.15 mmol) were added to dioxane (18 mL) in a single-necked
flask (50 mL), and then purged with nitrogen 3 times, the mixture
was heated to 80.degree. C. and reacted for 5 hours. The reaction
solution was cooled, added with ethyl acetate (100 mL), and washed
with water (100 mL) and brine (100 mL). The organic phase was
dried, filtered, concentrated, and purified by preparative silica
gel plate (ethyl acetate/petroleum ether: 1/4) to yield a yellow
solid product.
Step 2: Synthesis of methyl
2-(1-(((3-(4-cyanophenyl)pyridin-4-yl)thio)
methyl)cyclopropyl)acetate (20-c)
[0125] Methyl 2-(1-(mercaptomethyl)cyclopropyl)acetate (840 mg,
5.25 mmol), potassium carbonate (1.45 g, 10.5 mmol) and
4-(4-chloropyridin-3-yl) benzonitrile (450 mg, 2.1 mmol) were
dissolved in dimethyl formamide (20 mL) in a single-necked flask
(50 mL), the mixture was heated to 130.degree. C. and reacted for
0.5 hour. The reaction solution was cooled, added with ethyl
acetate (100 ml), and washed with water (100 ml) and brine
(100.times.3 mL). The organic phase was dried, filtered,
concentrated, and purified by preparative silica gel plate (ethyl
acetate/petroleum ether: 1/2) to yield a yellow oily product.
Step 3: Synthesis of
2-(1-(((3-(4-cyanophenyl)pyridin-4-yl)thio)methyl)
cyclopropyl)acetic acid (20)
[0126] Methyl
2-(1-(((3-(4-cyanophenyl)pyridin-4-yl)thio)methyl)cyclopropyl)
acetate (67 mg, 0.2 mmol) and aqueous solution of sodium hydroxide
(0.5 mL, 0.5 mmol, 1 M) were added to methanol (3 mL) in a
single-necked flask (50 mL), and the mixture was reacted at room
temperature for 5 hours. The reaction solution was adjusted to pH=3
with concentrated hydrochloric acid, concentrated and purified by
preparative reverse-phase chromatography to yield a white solid
product.
[0127] LC-MS (ES, m/z): 325 [M+H].sup.+; H-NMR (400 MHz,
CDCl.sub.3, ppm): .delta. 8.42 (s, 1H), 8.24 (s, 1H), 7.75 (d,
J=8.4 Hz, 2H), 7.53 (d, J=8.4 Hz, 2H), 7.35-7.33 (m, 1H), 3.19 (s,
2H), 2.38 (s, 2H), 0.62-0.60 (m, 4H).
Example 13: Synthesis of Compound 17
##STR00049##
[0128] Step 1: Synthesis of methyl
1-(hydroxymethyl)cyclobutanecarboxylate (17-b)
[0129] 1-(hydroxymethyl)cyclobutanecarboxylic acid (390 mg, 3 mmol)
was dissolved in methanol (20 mL) in a single-necked flask (50 mL),
thionyl chloride (1.7 g, 15 mmol) was added thereto at 0.degree.
C., then the mixture was stirred at 65.degree. C. for 4 hours. The
reaction solution was cooled, concentrated, and extracted by
addition of ethyl acetate (50 mL), and washed with aqueous solution
of sodium bicarbonate (20 mL) and brine (10 mL). The organic phase
was dried, filtered and concentrated to yield a yellow oily
product.
Step 2: Synthesis of methyl 1-(((methylsulfonyl)oxy)methyl)
cyclobutanecarboxylate (17-c)
[0130] 17-b (124 mg, 0.86 mmol) was dissolved in dichloromethane (6
mL) in a single-necked flask (50 mL), N,N-diisopropylethylamine
(332 mg, 2.6 mmol) and methanesulfonyl chloride (137 mg, 1.2 mmol)
were added thereto at 0.degree. C., then the mixture was stirred at
room temperature for 4 hours. The reaction solution was washed with
water (20 mL) and aqueous solution of sodium bicarbonate (15 mL).
The organic phase was dried, filtered and concentrated to yield a
yellow oily product.
Step 3: Synthesis of methyl
1-(((3-(4-cyanonaphthalen-1-yl)pyridin-4-yl)
thio)methyl)cyclobutanecarboxylate (17-d)
[0131] 1-b (400 mg, 1.5 mmol) was dissolved in
N,N-dimethylformamide (10 mL) in a single-necked flask (50 mL),
sodium sulfide (234 mg, 4.5 mmol) was added thereto, and the
mixture was reacted at 130.degree. C. for 2 hours. The reaction
solution was cooled and added with water (30 mL), after that, the
reaction solution was adjusted to pH=4 with 1 N aqueous solution of
hydrochloric acid, added with ethyl acetate (50 mL), then organic
phase was washed with water (30 mL) and brine (30 mL). The organic
phase was dried, filtered, and concentrated to yield a yellow solid
product.
[0132] The above yellow solid product (100 mg, 0.38 mmol),
potassium carbonate (210 mg, 1.52 mmol) and 17-c (200 mg, 0.9 mmol)
were added to N,N-dimethylformamide (10 mL) in a single-necked
flask (50 mL), and the mixture was reacted at 60.degree. C. for 2
hours. The reaction solution was cooled, added with ethyl acetate
(50 mL), and washed with water (30 mL) and brine (30 mL). The
organic phase was dried, filtered, concentrated and purified by
preparative silica gel plate (ethyl acetate/petroleum ether: 1/1)
to yield a yellow solid product.
Step 4: Synthesis of
1-(((3-(4-cyanonaphthalen-1-yl)pyridin-4-yl)thio)
methyl)cyclobutanecarboxylic acid (17)
[0133] 17-d (64 mg, 0.165 mmol) and 1 M of aqueous solution of
lithium hydroxide (0.82 mL, 0.82 mmol) were added to
tetrahydrofuran (4 mL) in a single-necked flask (50 mL), and the
mixture was reacted at room temperature for 36 hours. The reaction
solution was adjusted to pH=4 with 1 N of hydrochloric acid, then
added with ethyl acetate (50 mL), and washed with water (20 mL).
The organic phase was dried, filtered, concentrated and purified by
preparative silica gel plate (ethyl acetate/petroleum ether: 2/1)
to yield a white solid product.
[0134] LC-MS (ES, m/z): 375 [M+H].sup.+. H-NMR (400 MHz,
DMSO-d.sub.6, ppm): .delta. 12.42 (s, 1H), 8.56 (d, J=5.2 Hz, 1H),
8.18-8.26 (m, 3H), 7.83 (m, 1H), 7.63 (m, 2H), 7.55 (d, J=7.2 Hz,
1H), 7.44 (d, J=8.8 Hz, 1H), 3.42 (s, 2H), 2.20 (m, 2H), 1.86 (m,
4H).
Example 14: Synthesis of Compound 7
##STR00050##
[0136] Compound 7 was synthesized by a method similar to that in
Example 13, except that 1-(hydroxymethyl)cyclobutanecarboxylic acid
was replaced with the corresponding compound in step 1.
[0137] LC-MS (ES, m/z): 361 [M+H].sup.+. H-NMR (300 MHz,
CD.sub.3OD, ppm): .delta. 2.20-2.29 (m, 2H), 2.85-2.95 (m, 2H),
3.20-3.23 (m, 1H), 4.21-4.25 (m, 1H), 7.59-7.71 (m, 4H), 7.82-7.87
(t, J=7.5 Hz, 1H), 8.16-8.18 (d, J=7.2 Hz, 1H), 8.33-8.35 (d, J=8.4
Hz, 1H), 8.47 (m, 1H), 8.64 (m, 1H).
Example 15: Synthesis of Compound 16
##STR00051##
[0139] Compound 16 was synthesized by a method similar to that in
Example 13, except that 1-(hydroxymethyl)cyclobutanecarboxylic acid
was replaced with the corresponding compound in step 1.
[0140] LC-MS (ES, m/z): 389 [M+H].sup.+. H-NMR (400 MHz, DMSO-d6,
ppm): .delta. 8.55 (br s, 1H), 8.18-8.24 (m, 3H), 7.83 (br s, 1H),
7.55-7.59 (m, 3H), 7.44 (br s, 1H), 3.24 (br s, 2H), 1.87 (br s,
2H), 1.54-1.50 (m, 6H).
Example 16: Synthesis of Compound 11
##STR00052##
[0141] Step 1: Synthesis of (1-((ethoxycarbonyl)methyl)cyclopropyl)
methyl methanesulfonate (11-b)
[0142] Ethyl 2-(1-(hydroxymethyl)cyclopropyl)acetate (288 mg, 2
mmol) and triethylamine (404 mg, 4 mmol) were dissolved in
dichloromethane (3 mL) in a single-necked flask (50 mL),
methanesulfonyl chloride (342 mg, 3 mmol) was added thereto under
ice water bath condition, then the mixture was held at room
temperature and reacted for 3 hours. Then, the reaction solution
was added with ethyl acetate (50 mL), and washed with water (50 mL)
and brine (50 mL). The organic phase was dried, filtered, and
concentrated to yield a yellow oily product.
Step 2: Synthesis of ethyl
2-(1-((2-bromophenylthio)methyl)cyclopropyl) acetate (11-c)
[0143] (1-((ethoxycarbonyl)methyl)cyclopropyl)methyl
methanesulfonate (340 mg, 1.5 mmol), anhydrous potassium carbonate
(242 mg, 1.75 mmol) and 2-bromobenzenethiol (235 mg, 1.25 mmol)
were dissolved in dimethyl formamide (25 mL) in a single-necked
flask (50 mL), and the mixture was reacted at room temperature for
12 hours. Then, the reaction solution was added with ethyl acetate
(100 ml), and washed 3 times with water (50 mL) and brine (50 mL).
The organic phase was dried, filtered, concentrated, and purified
by preparative silica gel plate (ethyl acetate/petroleum ether:
1/7) to yield a yellow oily product.
Step 3: Synthesis of ethyl
2-(1-((2-(4-cyanophenyl)phenylthio)methyl) cyclopropyl)acetate
(11-d)
[0144] Ethyl 2-(1-((2-bromophenylthio)methyl)cyclopropyl)acetate
(65 mg, 0.2 mmol), aqueous solution of sodium carbonate (0.4 mL,
0.8 mmol, 2 M), 4-cyanophenylboronic acid (30 mg, 0.2 mmol) and
tetrakis(triphenylphosphine)palladium (0) (23 mg, 0.02 mmol) were
added to dioxane (2 mL) in a single-necked flask (50 mL), and then
purged with nitrogen 3 times, the mixture was heated to 80.degree.
C. and reacted for 4 hours. Then, the reaction solution was added
with ethyl acetate (50 ml), washed with water (50 mL) and brine (50
mL). The organic phase was dried, filtered, concentrated, and
purified by preparative silica gel plate (ethyl acetate/petroleum
ether: 1/5) to yield a yellow oily product.
Step 4: Synthesis of 2-(1-((2-(4-cyanophenyl)phenylthio)methyl)
cyclopropyl)acetic acid (compound 11)
[0145] Ethyl
2-(1-((2-(4-cyanophenyl)phenylthio)methyl)cyclopropyl)acetate (50
mg, 0.14 mmol) and aqueous solution of sodium hydroxide (1 mL, 1
mmol, 1 M) were added to methanol (3 mL) in a single-necked flask
(50 mL), and the mixture was reacted at room temperature for 16
hours. Then, the reaction solution was adjusted to pH=3 with
concentrated hydrochloric acid, added with ethyl acetate (50 mL),
and washed with water (50 mL) and brine (50 mL). The organic phase
was dried, filtered, concentrated, and purified by preparative
silica gel plate (ethyl acetate/petroleum ether: 2/3) to yield a
colorless oily product.
[0146] LCMS (ES, m/z): 324 [M+H].sup.+. H-NMR (400 MHz, CDCl.sub.3,
ppm): .delta. 7.72 (d, J=8.0 Hz, 2H), 7.56 (d, J=8.0 Hz, 2H),
7.48-7.46 (m, 1H), 7.33-7.24 (m, 2H), 7.20-7.18 (m, 1H), 2.89 (s,
2H), 2.32 (s, 2H), 0.50-0.42 (m, 4H).
Example 17: Synthesis of Compound 21
##STR00053##
[0148] Compound 21 was synthesized by a method similar to that in
Example 16, except that 2-bromobenzenethiol was replaced with the
corresponding compound in step 2.
[0149] LC-MS (ES, m/z): 342 [M+H].sup.+; H-NMR: (400 MHz,
CDCl.sub.3, ppm): .delta. 7.73-7.70 (m, 2H), 7.55-7.50 (m, 3H),
7.06-6.95 (m, 2H), 2.79 (s, 2H), 2.28 (s, 2H), 0.45-0.37 (m,
4H).
Example 18: Synthesis of Compound 23
##STR00054##
[0150] Step 1: Synthesis of ethyl
2-(1-((2-(4-cyanophenyl)sulfonyl)methyl) cyclopropyl)acetate
(23-a)
[0151] Ethyl
2-(1-((4-cyanophenyl)phenylthio)methyl)cyclopropyl)acetate (90 mg,
0.25 mmol), and potassium monopersulfate triple salt (473 mg, 0.75
mmol) were dissolved in methanol/water (5 mL, 4/1) in a
single-necked flask (50 mL), then the mixture was reacted at room
temperature for 24 hours. The reaction solution was added with
ethyl acetate (100 mL), washed with water (50 mL) and brine
(50.times.3 mL). The organic phase was dried, filtered,
concentrated, and purified by silica gel plate (ethyl
acetate/petroleum ether: 1/2) to yield a colorless oily
product.
Step 2: Synthesis of 2-(1-((2-(4-cyanophenyl)sulfonyl)methyl)
cyclopropyl)acetic acid (23)
[0152] Ethyl
2-(1-((2-(4-cyanobiphenyl)sulfonyl)methyl)cyclopropyl)acetate (64
mg, 0.17 mmol), and aqueous solution of lithium hydroxide (0.4 mL,
0.4 mmol, 1 M) were added to methanol (3 mL) in a single-necked
flask (50 mL), then the mixture was reacted at room temperature for
24 hours. The reaction solution was adjusted to pH=3 with
concentrated hydrochloric acid and purified by preparative
reverse-phase chromatography to yield a white solid product.
[0153] LC-MS (ES, m/z): 354 [M-H].sup.-; H-NMR: (400 MHz,
DMSO-d.sub.6, ppm): .delta. 8.06 (d, J=7.6 Hz, 1H), 7.86 (d, J=8.0
Hz, 2H), 7.80-7.77 (m, 1H), 7.73-7.69 (m, 1H), 7.53 (d, J=8.4 Hz,
2H), 7.38 (d, J=7.2 Hz, 1H), 3.13 (s, 2H), 2.17 (s, 2H), 0.35-0.33
(m, 4H).
Example 19: Synthesis of
2-(3-(1-cyanonaphthalen-4-yl)pyridin-4-ylamino)acetic acid
(compound 24)
##STR00055##
[0155] 4-(4-chloropyridin-3-yl)naphtha-1-carbonitrile (53 mg, 0.2
mmol), 2-glycine (37 mg, 0.5 mmol) and phenol (113 mg, 1.2 mmol)
were added in turn into a pipe sealing reaction bottle, the mixture
was heated to 120 degrees and reacted overnight. Then, the reaction
solution was cooled to room temperature, added with ether and
filtered. Filter cake was purified by preparative reverse-phase
chromatography to yield a white solid product.
[0156] LCMS (ES, m/z): 304 [M+H].sup.+. H-NMR: (400 MHz,
CD.sub.3OD, ppm): .delta. 8.33-8.31 (m, 2H), 8.19-8.15 (m, 2H),
7.84-7.76 (m, 2H), 7.70-7.67 (m, 2H), 7.08 (d, J=7.2 Hz, 1H), 4.06
(s, 2H).
Example 20: Synthesis of Compound 25
##STR00056##
[0158] Compound 25 was synthesized by a method similar to that in
Example 19, except that glycine was replaced with the corresponding
compound in the step.
[0159] LC-MS (ES, m/z): 332 [M+H].sup.+; H-NMR (400 MHz,
CD.sub.3OD, ppm): .delta. 8.19-8.13 (m, 4H), 7.85-7.67 (m, 4H),
7.17 (d, J=7.2 Hz, 1H), 1.53 (s, 3H), 1.52 (s, 3H).
Example 21: Synthesis of Compound 26
##STR00057##
[0161] Compound 26 was synthesized by a method similar to that in
Example 19, except that glycine was replaced with the corresponding
compound in the step.
[0162] LC-MS (ES, m/z): 318 [M+H].sup.+; H-NMR: (400 MHz,
DMSO-d.sub.6, ppm): .delta. 8.23-8.17 (m, 3H), 7.90 (s, 1H),
7.83-7.79 (m, 1H), 7.65-7.62 (m, 1H), 7.57-7.52 (m, 2H), 6.75 (d,
J=6.4 Hz, 1H), 5.46 (s, 1H), 3.25-3.23 (m, 2H), 2.29-2.28 (m,
2H).
Example 22: Synthesis of Compound 27
##STR00058##
[0164] Compound 27 was synthesized by a method similar to that in
Example 19, except that glycine was replaced with the corresponding
compound in the step.
[0165] LC-MS (ES, m/z): 332 [M+H].sup.+; H-NMR: (400 MHz,
DMSO-d.sub.6, ppm): .delta. 8.20-8.17 (m, 3H), 7.88 (s, 1H),
7.83-7.79 (m, 1H), 7.65-7.62 (m, 1H), 7.57-7.53 (m, 2H), 6.73 (d,
J=6.0 Hz, 1H), 5.54 (s, 1H), 3.01 (d, J=6.4 Hz, 2H), 2.10 (d, J=7.2
Hz, 2H), 1.60-1.56 (m, 2H).
Example 23: Synthesis of Compound 28
##STR00059##
[0167] 4-(4-chloropyridin-3-yl)benzonitrile (67 mg, 0.3 mmol),
glycine (56 mg, 0.75 mmol) and phenol (169 mg, 1.8 mmol) were added
in turn into a pipe sealing reaction bottle, the mixture was heated
to 120 degrees and reacted overnight. The reaction solution was
cooled to room temperature, added with ether and filtered. Filter
cake was purified by preparative reverse-phase chromatography to
yield a white solid product.
[0168] LCMS (ES, m/z): 254 [M+H].sup.+; H-NMR: (400 MHz,
CD.sub.3OD, ppm): .delta. 8.47 (br s, 2H), 8.15 (d, J=6.8 Hz, 1H),
8.00 (s, 1H), 7.90 (d, J=8.0 Hz, 1H), 7.71 (d, J=8.0 Hz, 1H), 6.80
(d, J=6.8 Hz, 1H), 3.81 (s, 2H).
Example 24: Synthesis of Compound 29
##STR00060##
[0169] Step 1: Synthesis of ethyl
2-(1-(((3-bromopyridin-4-yl)oxy)methyl) cyclopropyl)acetate
(29-a)
[0170] 3-bromopyridin-4-ol (500 mg, 2.9 mmol) was dissolved in
tetrahydrofuran (10 mL) in a three-necked flask (100 mL), ethyl
2-(1-hydroxymethyl)cyclopropyl)acetate (428 mg, 2.9 mmol),
triphenylphosphine (909 mg, 3.5 mmol) and diethylazodicarboxylate
(609 mg, 3.5 mmol) were added thereto in turn at 0.degree. C. under
the protection of nitrogen, and then the mixture was heated to room
temperature and allowed to carry out Mitsunobu Reaction for 16
hours. The reaction solution was directly concentrated, and
purified by preparative silica gel plate (ethyl acetate/petroleum
ether: 2/1) to yield a white solid.
Step 2: Synthesis of ethyl
2-(1-(((3-(4-cyanophenyl)pyridin-4-yl)oxy)
methyl)cyclopropyl)acetate(29-b)
[0171] Ethyl
2-(1-(((3-bromopyridin-4-yl)oxy)methyl)cyclopropyl)acetate (29-a)
(90 mg, 0.29 mmol), aqueous solution of sodium carbonate (1 mL, 2
mmol, 2 M), 4-cyanophenylboronic acid (43 mg, 0.29 mmol) and
tetrakis(triphenylphosphine)palladium (0) (33 mg, 0.03 mmol) were
added to dioxane (3 mL) in a single-necked flask (50 mL), purged
with nitrogen 3 times, and then the mixture was heated to
80.degree. C. and reacted for 12 hours. The reaction solution was
cooled to room temperature, added with ethyl acetate (50 mL), and
washed with water (50 mL) and brine (50 mL). The organic phase was
dried, filtered, concentrated, and purified by preparative silica
gel plate (dichloromethane/methanol: 20/1) to yield a yellow oily
product.
Step 3: Synthesis of
2-(1-(((3-(4-cyanophenyl)pyridin-4-yl)oxy)methyl)
cyclopropyl)acetic acid (29)
[0172] Ethyl
2-(1-(((3-(4-cyanophenyl)pyridin-4-yl)oxy)methyl)cyclopropyl)
acetate (60 mg, 0.18 mmol) and lithium hydroxide (41 mg, 0.97 mmol)
were added to tetrahydrofuran/water (3 mL/1 mL) in a single-necked
flask (50 mL), and the mixture was reacted at room temperature for
16 hours. The reaction solution was adjusted to pH=4 with
concentrated hydrochloric acid, added with ethyl acetate (50 mL),
and washed with water (50 mL) and brine (50 mL). The organic phase
was dried, filtered, concentrated, and purified by preparative
silica gel plate (dichloromethane/methanol: 20/1) to yield a white
solid product.
[0173] LC-MS (ES, m/z): 309 [M+H].sup.+. H-NMR: (400 MHz,
CDCl.sub.3, ppm): .delta. 8.05 (d, J=2.0 Hz, 1H), 7.83-7.75 (m,
5H), 6.57-6.55 (m, 1H), 4.07 (s, 2H), 2.22 (s, 2H), 0.84-0.82 (m,
2H), 0.73-0.72 (m, 2H).
Example 25: Synthesis of Compound 30
##STR00061##
[0175] Compound 30 was synthesized by a method similar to that in
Example 24, except that (4-cyanophenyl)boronic acid was replaced
with the corresponding compound in step 2.
[0176] LC-MS (ES, m/z): 359 [M+H].sup.+; H-NMR (400 MHz,
CD.sub.3OD, ppm): .delta. 8.22 (d, J=8.4 Hz, 1H), 8.05 (d, J=7.6
Hz, 1H), 8.02-8.00 (m, 2H), 7.82 (d, J=8.4 Hz, 1H), 7.75 (m, 1H),
7.62 (m, 1H), 7.54 (d, J=7.6 Hz, 1H), 6.61 (d, J=7.2 Hz, 1H),
4.12-4.09 (m, 2H), 2.25 (s, 2H), 0.81 (m, 2H), 0.71 (m, 2H).
Example 26: Synthesis of Compound 31
##STR00062##
[0177] Step 1: Synthesis of methyl
2-(3-bromopyridin-4-yloxy)-2-methylpropanoate (31-a)
[0178] Methyl 2-bromo-2-methylpropanoate (724 mg, 4 mmol),
potassium carbonate (828 mg, 6 mmol) and 3-bromo-4-hydroxypyridine
(348 mg, 2 mmol) were dissolved in dimethyl formamide (20 mL) in a
single-necked flask (50 mL), then the mixture was reacted at
60.degree. C. for 12 hours. The reaction solution was cooled to
room temperature, added with ethyl acetate (100 mL), and washed
with water (50 mL) and brine (50.times.3 mL). The organic phase was
dried, filtered, concentrated, and purified by silica gel column
(dichloromethane/methanol: 50/1-20/1) to yield a colorless oily
product.
Step 2: Synthesis of
2-(3-(4-cyanophenyl)pyridin-4-yloxy)-2-methylpropanoic acid
(31)
[0179] Methyl 2-(3-bromopyridin-4-yloxy)-2-methylpropanoate (109
mg, 0.4 mmol), aqueous solution of sodium carbonate (0.8 mL, 1.6
mmol, 2 M), 4-cyanophenylboronic acid (59 mg, 0.4 mmol) and
tetrakis(triphenylphosphine)palladium (0) (46 mg, 0.04 mmol) were
added to dioxane (2.4 mL) in a single-necked flask (50 mL), purged
with nitrogen 3 times, and then the mixture was heated to
80.degree. C. and reacted for 12 hours. The reaction solution was
cooled to room temperature, adjusted to pH=4, added with ethyl
acetate (100 mL), and washed with water (100 mL) and brine (100
mL). The organic phase was dried, filtered, concentrated, and
purified by preparative reverse-phase chromatography to yield a
white solid product.
[0180] LC-MS (ES, m/z): 283 [M+H].sup.+; H-NMR (400 MHz,
CD.sub.3OD, ppm): .delta. 8.35-8.30 (m, 2H), 7.82-7.77 (m, 4H),
6.98-6.97 (m, 1H), 1.59 (s, 6H).
Example 27: Synthesis of Compound 10
##STR00063##
[0181] Step 1: Synthesis of
1-(4-chloropyridin-3-yl)isoquinoline-4-carbonitrile (10-a)
[0182] In a three-necked flask (100 mL), under the protection of
nitrogen, 1-chloroisoquinoline-4-carbonitrile (450 mg, 2.39 mmol)
was dissolved in toluene (30 mL) and water (3 mL), and
4-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
(687 mg, 2.87 mmol), sodium carbonate (761 mg, 7.18 mmol) and
palladium catalyst tetrakis(triphenylphosphine)palladium (0) (138
mg, 0.12 mmol) were added thereto, then the mixture was heated to
95.degree. C. and reacted for 2 hours. The reaction solution was
cooled, and the reaction was quenched by addition of 50 mL of ice
water, then the reaction solution was extracted with ethyl acetate
(100 mL, 3 times), reversely washed with saturated brine (100 mL, 3
times), dried over anhydrous sodium sulfate, rotary evaporated, and
purified by column chromatography (petroleum ether/ethyl
acetate=10:1-petroleum ether/ethyl acetate/dichloromethane=1:1:1),
to yield an off-white solid product.
Step 2: Synthesis of methyl
2-((3-(4-cyanoisoquinolin-1-yl)pyridin-4-yl)
thio)-2-methylpropanoate (10-b)
[0183] In a three-necked flask (100 mL), under the protection of
nitrogen, 1-(4-chloropyridin-3-yl)isoquinoline-4-carbonitrile (110
mg, 0.41 mmol) obtained in step 2 was dissolved in dimethyl
formamide (20 mL), the solution was then added with sodium sulfide
(194 mg, 2.49 mmol), the mixture was heated to 130.degree. C. and
reacted for about 1.5 hours, then cooled to room temperature,
followed by addition of anhydrous potassium carbonate (286 mg, 2.07
mmol) and then methyl 1-(bromomethyl)cyclopropanecarboxylate (224
mg, 1.24 mmol), the mixture was then heated to 130.degree. C. and
allowed to further react for about 1.5 hours until the reaction was
completed. The reaction solution was cooled and poured into 50 mL
of ice water to quench the reaction, then the reaction solution was
extracted with ethyl acetate (50 mL, 3 times), the organic phase
was dried over anhydrous sodium sulfate, rotatory evaporated, and
purified through column chromatography (petroleum ether/ethyl
acetate=10:1.about.petroleum ether/ethyl
acetate/dichloromethane=1:1:1), to yield a yellow oily product.
Step 3: Synthesis of
2-((3-(4-cyanoisoquinolin-1-yl)pyridin-4-yl)thio)-2-methylpropanoic
acid (compound 10)
[0184] methyl
2-((3-(4-cyanoisoquinolin-1-yl)pyridin-4-yl)thio)-2-methylpropanoate
(60 mg, 0.17 mmol) obtained in step 3, lithium hydroxide (12 mg,
0.50 mmol), tetrahydrofuran (24 mL) and water (8 mL) were added
into a three-necked flask (100 mL) under the protection of N.sub.2,
the mixture was reacted at room temperature overnight.
Tetrahydrofuran was removed by concentration; aqueous phase was
extracted 3 times with dichloromethane (50 mL) and collected. Then
the aqueous phase was adjusted to pH=4-5 with 2 N of hydrochloric
acid regulating system and extracted with dichloromethane (100 mL,
3 times); the organic phase was combined, then dried with sodium
sulfate, and rotatory evaporated. The crude product was subject to
high pressure to yield a white solid product.
[0185] LC-MS (ES, m/z): 350 [M+H].sup.+; H-NMR (300 MHz,
CD.sub.3OD, ppm): .delta. 1.55 (s, 6H), 7.83-7.95 (m, 3H),
8.09-8.14 (m, 1H), 8.33-8.36 (d, J=8.4 Hz, 1H), 8.67-8.74 (m, 2H),
9.09 (s, 1H).
Example 28: Synthesis of Compound 8
##STR00064##
[0186] Step 1: Synthesis of
1-(4-chloropyridin-3-yl)-1H-indole-3-carbonitrile (8-a)
[0187] In a three-necked flask (100 mL), under the protection of
N.sub.2, copper acetate (1800 mg, 9.9 mmol) and pyridine (1200 mg,
15.1 mmol) were sequentially added to 1H-indole-3-carbonitrile (700
mg, 4.9 mmol) and
4-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
(1300 mg, 5.4 mmol) which were dissolved in dimethyl formamide (50
mL), then the mixture was stirred at room temperature.
4-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
(700 mg, 3.0 mmol) was replenished every 4 hours, 5 times in total.
The reaction solution was poured into 100 mL of ice water to make
the reaction quenched, then the reaction solution was extracted
with ethyl acetate (150 mL, 3 times) and washed with saturated
brine (150 mL, 3 times). The organic phase was dried over anhydrous
sodium sulfate, rotatory evaporated and subject to high pressure to
yield a white solid.
Step 2: Synthesis of methyl
2-((3-(3-cyano-1H-indol-1-yl)pyridin-4-yl) thio)-2-methylpropanoate
(8-b)
[0188] In a three-necked flask (100 mL), under the protection of
N.sub.2, 1-(4-chloropyridin-3-yl)-1H-indole-3-carbonitrile (100 mg,
0.39 mmol) obtained in step 1 was dissolved in dimethyl formamide
(50 mL), then sodium sulfide (185 mg, 2.37 mmol) was added thereto,
the mixture was heated to 130.degree. C. and reacted for about 1
hour, then cooled to room temperature, followed by addition of
anhydrous potassium carbonate (273 mg, 1.98 mmol) and then methyl
1-(bromomethyl)cyclopropanecarboxylate (213 mg, 1.18 mmol), the
mixture was heated to 130.degree. C. and allowed to further react
for about 1.5 hour until the reaction was completed. The reaction
solution was cooled and poured into 100 mL ice water, thus the
reaction was quenched. The reaction solution was extracted with
ethyl acetate (150 mL, 3 times), the organic phase was dried over
anhydrous sodium sulfate, rotatory evaporated and subject to high
pressure to yield a light yellow solid.
Step 3: Synthesis of
2-((3-(3-cyano-1H-indol-1-yl)pyridin-4-yl)thio)-2-methylpropanoic
acid (compound 8)
[0189] Methyl
2-((3-(3-cyano-1H-indol-1-yl)pyridin-4-yl)thio)-2-methylpropanoate
(50 mg, 0.10 mmol) obtained in step 2, lithium hydroxide (11 mg,
0.40 mmol), tetrahydrofuran (24 mL) and water (8 mL) were added
into a three-necked flask (50 mL) under the protection of N.sub.2,
and reacted at room temperature overnight. Tetrahydrofuran was
removed by concentration; aqueous phase was extracted 3 times with
dichloromethane (50 mL) and collected. Then the aqueous phase was
adjusted to pH=4-5 with 2N of hydrochloric acid regulating system
and extracted with dichloromethane (100 mL, 3 times); the organic
phase was combined, dried over sodium sulfate and then rotatory
evaporated. The crude product was subject to high pressure to yield
a light yellow solid.
[0190] LC-MS (ES, m/z): 338 [M+H].sup.+; H-NMR (300 MHz,
CD.sub.3OD, ppm):.delta.1.54 (s, 6H), 7.13-16 (m, 1H), 7.36-43 (m,
2H), 7.76-7.82 (m, 2H), 8.13 (s, 1H), 8.63-8.68 (m, 2H).
Example 29: Synthesis of Compound 9
##STR00065##
[0192] Compound 9 was synthesized by a method similar to that in
Example 28, except that 1H-indole-3-carbonitrile was replaced with
the corresponding compound in step 1.
[0193] LC-MS (ES, m/z): 354 [M+H].sup.+; H-NMR (300 MHz,
CD.sub.3OD, ppm): .delta. 1.12-1.14 (m, 4H), 1.51 (s, 6H),
2.32-2.41 (m, 1H), 7.16-7.19 (d, J=8.4 Hz, 1H), 7.24-7.28 (m, 1H),
7.43-7.48 (m, 1H), 7.685 (m, 1H), 7.88-7.91 (d, J=8.1 Hz, 1H), 8.50
(br s, 2H).
Example 30: Synthesis of Compound 22
##STR00066##
[0194] Step 1: Synthesis of methyl
2-(1-((3-bromopyridin-4-ylthio)methyl) cyclopropyl)acetate
(22-a)
[0195] In a single-necked flask (50 mL), methyl
2-(1-(mercaptomethyl) cyclopropyl)acetate (2 g, 12.5 mmol),
potassium carbonate (3.45 g, 25 mmol), and 3-bromo-4-chloropyridine
(955 mg, 5 mmol) were dissolved in dimethyl formamide (30 mL), the
mixture was heated to 130.degree. C. and reacted for 2 hours. The
reaction solution was cooled to room temperature, added with ethyl
acetate (100 ml), and washed with water (100 mL) and brine (100 mL,
3 times). The organic phase was dried, filtered, concentrated and
purified by silica gel column (ethyl acetate/petroleum ether:
1/10-1/6) to yield a yellow solid product.
Step 2: Synthesis of methyl
2-(1-((3-(4-(hydroxymethyl)phenylpyridin-4-yl)thio)methyl)cyclopropyl)ace-
tate (22-b)
[0196] Methyl
2-(1-((3-bromopyridin-4-ylthio)methyl)cyclopropyl)acetate (160 mg,
0.5 mmol), aqueous solution of sodium carbonate (1 mL, 2 mmol, 2
M), 4-hydroxymethylphenylboric acid (76 mg, 0.5 mmol) and
tetrakis(triphenylphosphine)palladium(0) (60 mg, 0.05 mmol) were
added to dioxane (3 mL) in a single-necked flask (50 mL), purged
with nitrogen 3 times, and then the mixture was heated to
80.degree. C. and reacted for 12 hours. The reaction solution was
cooled to room temperature, added with ethyl acetate (100 ml), and
washed with water (100 mL) and brine (100 mL). The organic phase
was dried, filtered, concentrated and purified by preparative
silica gel plate (ethyl acetate/petroleum ether: 1/1) to yield a
yellow solid product.
Step 3: Synthesis of
2-(1-((3-(4-(hydroxymethyl)phenyl)pyridin-4-ylthio)
methyl)cyclopropyl)acetic acid (22)
[0197] In a single-necked flask (50 mL), methyl
2-(1-(((3-(4-(hydroxymethyl)
phenyl)pyridin-4-yl)thio)methyl)cyclopropyl)acetate (140 mg, 0.4
mmol) and aqueous solution of aluminum hydroxide (0.8 mL, 0.8 mmol,
1 M) were added to methanol (3 mL), then the mixture was reacted at
room temperature for 2 hours. The reaction solution was adjusted to
pH=3 with concentrated hydrochloric acid, concentrated and purified
by preparative silica gel plate (dichloromethane/methanol: 10/1) to
yield a white solid product.
[0198] LC-MS (ES, m/z): 330 [M+H].sup.+; H-NMR (400 MHz,
DMSO-d.sub.6, ppm): .delta. 8.35 (d, J=6.4 Hz, 1H), 8.19 (s, 1H),
7.42-7.33 (m, 5H), 5.25 (s, 1H), 4.50 (d, J=6.4 Hz, 2H), 3.12 (s,
2H), 2.24 (s, 2H), 0.52-0.50 (m, 4H).
Example 31: Synthesis of Compound 36
##STR00067##
[0199] Step 1: Synthesis of 3-(4-bromophenyl)oxetan-3-ol (36-a)
[0200] In a three-necked flask (50 mL), under the protection of
nitrogen, 1,4-dibromobenzene (600 mg, 2.55 mmol) was dissolved in
tetrahydrofuran (15 mL), then the solution was cooled to 78.degree.
C. and added with n-butyllithium (1.05 mL, 2.55 mmol, 2.5 M in
hexane). The mixture was reacted for 0.5 hour, then added dropwise
with oxetan-3-one (153 mg, 2.55 mmol), and the resulting mixture
was further reacted for 3 hours. The reaction was quenched with
saturated aqueous solution of ammonium chloride (20 mL), the
reaction solution was added with ethyl acetate (50 ml), then washed
with brine (10 mL). The organic phase was dried, filtered,
concentrated and purified by preparative silica gel plate
(petroleum ether/ethyl acetate: 3/1) to yield a white solid.
Step 2: Synthesis of
3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)oxetan-3-ol
(36-b)
[0201] 36-a (300 mg, 1.2 mmol), potassium acetate (323 mg, 3.3
mmol), bis(pinacolato)diboron (B.sub.2(pin).sub.2) (420 mg, 1.6
mmol), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium
(100 mg, 0.12 mmol) were added to 1,4-dioxane (15 mL) in a
single-necked flask (100 mL), purged with nitrogen 3 times, and
then the mixture was heated to 90.degree. C. and reacted for 16
hours. The reaction solution was cooled, added with ethyl acetate
(60 ml), and washed with water (40 mL). The organic phase was
dried, filtered, concentrated and purified by silica gel column
(petroleum ether/ethyl acetate: 3/1) to yield a white solid.
Step 3: Synthesis of methyl
2-(1-(((3-(4-(3-hydroxyoxetan-3-yl)phenyl)
pyridin-4-yl)thio)methyl)cyclopropyl)acetate (36-c)
[0202] 36-b (150 mg, 0.54 mmol), aqueous solution of sodium
carbonate (2.2 mL, 2.2 mmol, 1 M), 22-a (170 mg, 0.54 mmol), and
tetrakis(triphenylphosphine)palladium(0) (62 mg, 0.054 mmol) were
added to dioxane (8 mL) in a single-necked flask (100 mL), and then
purged with nitrogen 3 times. The mixture was heated to 80.degree.
C. and reacted for 16 hours. The reaction solution was cooled,
added with ethyl acetate (50 mL), and washed with water (40 mL).
The organic phase was dried, filtered, concentrated and purified by
preparative silica gel plate (petroleum ether/ethyl acetate: 1/1)
to yield a white solid.
Step 4: Synthesis of
2-(1-(((3-(4-(3-hydroxyoxetan-3-yl)phenyl)pyridin-4-yl)thio)methyl)cyclop-
ropyl)acetic acid (36)
[0203] Experimental process: In a single-necked flask (50 mL), 36-c
(70 mg, 0.18 mmol) and 1 M of aqueous solution of lithium hydroxide
(1 mL, 1 mmol) were added to tetrahydrofuran (3 mL), and the
mixture was reacted at room temperature for 16 hours. The reaction
solution was adjusted to pH=4 with 1 N of hydrochloric acid, added
with chloroform-d/isopropanol (30 mL/10 mL) and washed with brine
(50 mL). The organic phase was dried, filtered, concentrated and
purified by preparative reverse-phase chromatography to yield a
white solid product.
[0204] LC-MS (ES, m/z): 372 [M+H].sup.+; H-NMR (400 MHz, DMSO-d6,
ppm): .delta. 8.35 (d, J=5.6 Hz, 1H), 8.20 (s, 1H), 7.68 (m, 2H),
7.43 (m, 2H), 7.35 (d, J=5.2 Hz, 1H), 6.41 (br s, 1H), 4.77 (d,
J=6.4 Hz, 2H), 4.72 (d, J=6.8 Hz, 2H), 3.15 (s, 2H), 2.22 (s, 2H),
0.49 (m, 4H).
Example 32: Synthesis of Compound 35
##STR00068##
[0205] Step 1: Synthesis of
3-(4-(4-chloropyridin-3-yl)phenyl)oxetan-3-ol (35-a)
[0206] 36-b (200 mg, 0.72 mmol), aqueous solution of sodium
carbonate (1.4 mL, 2.8 mmol, 2 M), 3-bromo-4-chloropyridine (139
mg, 0.72 mmol), and tetrakis(triphenylphosphine)palladium(0) (83
mg, 0.072 mmol) were added to dioxane (6 mL) in a single-necked
flask (50 mL), purged with nitrogen 3 times, and then the mixture
was heated to 80.degree. C. and reacted for 16 hours. The reaction
solution was added with ethyl acetate (50 ml) and washed with water
(40 mL). The organic phase was dried, filtered, concentrated and
purified by preparative silica gel plate (ethyl acetate/petroleum
ether: 1/1) to yield a white solid product.
Step 2: Synthesis of
2-((3-(4-(3-hydroxyoxetan-3-yl)phenyl)pyridin-4-yl)thio)-2-methylpropanoi-
c acid (35)
[0207] In a single-necked flask (50 mL), 35-a (140 mg, 0.54 mmol)
and sodium sulfide (125 mg, 1.6 mmol) were added to dimethyl
formamide (6 mL), the mixture was heated to 130.degree. C. and
reacted for 1 hour. After being cooled, the reaction solution was
added with anhydrous potassium carbonate (220 mg, 1.6 mmol), methyl
2-bromoisobutyrate (289 mg, 1.6 mmol), and the resulting solution
was reacted for another 1 hour at 130.degree. C. The reaction
solution was added with ether (50 mL) and water (50 mL), aqueous
phase was adjusted to pH=4 with dilute hydrochloric acid (1 M),
then extracted by addition of chloroform-d/isopropanol (30 mL/10
mL), and washed with brine (20 mL). The organic phase was dried,
filtered, concentrated and preparatively purified to yield a white
solid product.
[0208] LC-MS (ES, m/z): 346 [M+H].sup.+; H-NMR (400 MHz, DMSO-d6,
ppm): .delta. 8.40 (d, J=5.6 Hz, 1H), 8.29 (s, 1H), 7.66 (m, 2H),
7.38 (m, 3H), 6.42 (br s, 1H), 4.77 (d, J=6.4 Hz, 2H), 4.71 (d,
J=6.8 Hz, 2H), 1.44 (s, 6H).
Example 33: Synthesis of Compound 34
##STR00069##
[0209] Step 1: Synthesis of methyl 4-(4-chloropyridin-3-yl)benzoate
(34-a)
[0210] 3-bromo-4-chloropyridine (764 mg, 4 mmol), aqueous solution
of sodium carbonate (8 mL, 16 mmol, 2 M),
4-(methoxycarbonyl)phenylboronic acid (860 mg, 4 mmol), potassium
acetate(392 mg, 4 mmol), and
tetrakis(triphenylphosphine)palladium(0) (164 mg, 0.2 mmol) were
added to dioxane (24 mL) in a single-necked flask (100 mL), purged
with nitrogen 3 times, then the mixture was heated to 80.degree. C.
and reacted for 12 hours. The reaction solution was added with
ethyl acetate (100 mL), and washed with water (100 mL) and brine
(100 mL). The organic phase was dried, filtered, concentrated and
purified by silica gel column (ethyl acetate/petroleum ether:
1/8-1/4) to yield a white solid product.
Step 2: Synthesis of
1-(4-(4-chloropyridin-3-yl)phenyl)cyclopropanol (34-b)
[0211] In a three-necked flask (50 mL), ethylmagnesium bromide (in
ether (1 M), 6.8 mL, 6.8 mmol) was slowly added dropwise to the
solution of 34-a (838 mg, 3.4 mmol) and titanium tetraisopropoxide
(0.85 g, 3.4 mmol) in toluene (30 mL) under room temperature
condition within 30 min, then the mixture was reacted for 1 hour.
The reaction solution was further added with titanium
tetraisopropoxide (0.85 g, 3.4 mmol) and ethylmagnesium bromide (1
M, in Et.sub.2O, 6.8 mL, 6.8 mmol) and reacted for another 0.5
hour. The reaction was quenched with water, extracted with ethyl
acetate (100 mL) and washed with water (100 mL) and brine (100 mL).
The organic phase was dried, filtered, concentrated and purified by
preparative reverse-phase chromatography to yield a white solid
product.
Step 3: Synthesis of
3-(4-(1-((tert-butyldimethylsilyl)oxy)cyclopropyl)
phenyl)-4-chloropyridine (34-c)
[0212] In a single-necked flask (50 mL), under ice bath condition,
the solution of 34-b (150 mg, 0.6 mmol) and imidazole (204 mg, 3
mmol) in dimethyl formamide (5 mL) was added with
tert-butyldimethylsilyl chloride (453 mg, 3 mmol), then the mixture
was reacted at room temperature for 2 hours. The reaction solution
was added with ethyl acetate (50 mL), and washed with water (50 mL)
and brine (50 mL). The organic phase was dried, filtered,
concentrated and purified by silica gel plate (ethyl
acetate/petroleum ether: 1/4) to yield a white solid product.
Step 4: Synthesis of methyl
2-(1-(((3-(4-(1-((tert-butyldimethylsilyl)oxy)
cyclopropyl)phenyl)pyridin-4-yl)thio)methyl)cyclopropyl)acetate
(34-d)
[0213] In a single-necked flask (50 mL), 34-c (70 mg, 0.2 mmol),
anhydrous potassium carbonate (138 mg, 1 mmol), and methyl
2-(1-(mercaptomethyl) cyclopropyl)acetate (80 mg, 0.5 mmol) were
dissolved in dimethyl formamide (3 mL), the mixture was heated to
130.degree. C. and reacted for 0.5 hour. The reaction solution was
added with ethyl acetate (100 mL), and washed with water (100 mL)
and brine (100.times.3 mL). The organic phase was dried, filtered,
concentrated and purified by preparative silica gel plate (ethyl
acetate/petroleum ether: 1/3) to yield a yellow oily product.
Step 5: Synthesis of 2-(1-(((3-(4-(1-(tert-butyldimethylsilyl)oxy)
cyclopropyl)phenyl)pyridin-4-yl)thio)methyl)cyclopropyl)acetic acid
(34-e)
[0214] In a single-necked flask (50 mL), 34-d (120 mg, 0.25 mmol)
and aqueous solution of lithium hydroxide (0.75 mL, 0.75 mmol, 1 M)
were added to methanol (3 mL), and reacted for 2 hours at room
temperature. The reaction solution was adjusted to pH=3 with
concentrated hydrochloric acid, and concentrated to dryness.
Step 6: Synthesis of
2-(1-(((3-(4-(1-hydroxycyclopropyl)phenyl)pyridin-4-yl)thio)methyl)cyclop-
ropyl)acetic acid (34)
[0215] In a single-necked flask (50 mL), 34-e (obtained in the last
step, 0.25 mmol) and tetrabutylammonium fluoride (TBAF) (0.5 mL,
0.5 mmol, 1 M) were added to tetrahydrofuran (3 mL), and reacted at
room temperature for 0.5 hour. The reaction solution was added with
ethyl acetate (50 mL) and washed with water (50 mL) and brine
(50.times.3 mL). The organic phase was dried, filtered,
concentrated and purified by preparative reverse-phase
chromatography to yield a white solid product.
[0216] LC-MS (ES, m/z): 356 [M+H].sup.+; H-NMR (400 MHz,
CD.sub.3OD, ppm): .delta. 8.48 (d, J=6.4 Hz, 1H), 8.35 (s, 1H),
7.93 (d, J=6.8 Hz, 1H), 7.48-7.42 (m, 4H), 3.38 (s, 2H), 2.36 (s,
2H), 1.28-1.25 (m, 2H), 1.12-1.09 (m, 2H), 0.66-0.65 (m, 4H).
Example 34: Synthesis of Compound 33
##STR00070##
[0217] Step 1: Synthesis of
4-chloro-3-(4-(1-((tetrahydro-2H-pyran-2-yl)oxy)
cyclopropyl)phenyl)pyridine (33-a)
[0218] In a single-necked flask (50 mL), the solution of 34-b (50
mg, 0.2 mmol) and p-toluene sulfonic acid (PTSA) (7 mg, 0.04 mmol)
in dichloromethane (5 mL) was added with dihydropyran (DHP) (33 mg,
0.4 mmol), and the mixture was reacted at room temperature for 12
hours. The reaction solution was added with ethyl acetate (50 ml),
and then washed with water (50 mL) and brine (50 mL). The organic
phase was dried, filtered, concentrated and purified by silica gel
plate (ethyl acetate/petroleum ether: 1/4) to yield a colorless
oily product.
Step 2: Synthesis of methyl
2-methyl-2-((3-(4-(1-((tetrahydro-2H-pyran-2-yl)oxy)cyclopropyl)phenyl)py-
ridin-4-yl)thio)propanoate (33-b)
[0219] In a single-necked flask (50 mL), 33-a (37 mg, 0.11 mmol)
and sodium sulfide (26 mg, 0.33 mmol) were dissolved in dimethyl
formamide (4 mL), the mixture was heated to 130.degree. C. and
reacted for 1 hour. After being cooled, the mixture was added with
anhydrous potassium carbonate (76 mg, 0.55 mmol) and methyl
2-bromoisobutyrate (60 mg, 0.33 mmol), then heated to 50.degree. C.
and the resulting mixture was allowed to further react for 1 hour.
The reaction solution was added with ethyl acetate (50 mL), and
washed with water (50 mL) and brine (50 mL). The organic phase was
dried, filtered, concentrated and purified by preparative silica
gel plate (ethyl acetate/petroleum ether: 1/3) to yield a yellow
oily substance.
Step 3: Synthesis of
2-methyl-2-((3-(4-(1-((tetrahydro-2H-pyran-2-yl)
oxy)cyclopropyl)phenyl)pyridin-4-yl)thio)propanoic acid (33-c)
[0220] In a single-necked flask (50 mL), 33-b (35 mg, 0.08 mmol)
and the aqueous solution of lithium hydroxide (0.24 mL, 0.24 mmol,
1 M) were added to methanol (3 mL), and reacted for 6 hours at room
temperature. The reaction solution was adjusted to pH=5 with
concentrated hydrochloric acid, and then concentrated to yield a
yellow oily substance.
Step 4: Synthesis of
2-((3-(4-(1-hydroxycyclopropyl)phenyl)pyridin-4-yl)
thio)-2-methylpropanoic acid (33)
[0221] In a single-necked flask (50 mL), 33 (25 mg, 0.06 mmol) and
p-toluene sulfonic acid (2 mg, 0.01 mmol) were added to methanol (3
mL), and reacted for 1 hour at room temperature. The reaction
solution was added with ethyl acetate (50 ml), and washed with
water (50 mL) and brine (50.times.3 mL). The organic phase was
dried, filtered, concentrated and purified by preparative silica
gel plate (dichloromethane/methanol: 8/1) to yield a light yellow
solid product.
[0222] LC-MS (ES, m/z): 330 [M+H].sup.+; H-NMR (400 MHz, DMSO-d6,
ppm): .delta. 13.17 (s, 1H), 8.41 (d, J=6.4 Hz, 1H), 8.29 (s, 1H),
7.33-7.25 (m, 5H), 5.99 (s, 1H), 1.44 (s, 6H), 1.13-1.12 (m, 2H),
0.99-0.98 (m, 2H).
Example 35: Synthesis of Compound 37
##STR00071##
[0223] Step 1: Synthesis of 4'-chloro-[2,3'-bipyridine]-5-nitrile
(37-a)
[0224]
4-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
(400 mg, 1.64 mmol), the aqueous solution of sodium carbonate (3.3
mL, 6.6 mmol, 2 M), 6-bromopyridin-3-carbonitrile (300 mg, 1.64
mmol) and tetrakis(triphenylphosphine)palladium (0) (180 mg, 0.16
mmol) were added to dioxane (10 mL) in a single-necked flask (50
mL), and purged with nitrogen 3 times, and then the mixture was
heated to 80.degree. C. and reacted for 5 hours. The reaction
solution was added with ethyl acetate (50 mL), and washed with
water (50 mL) and brine (50 mL). The organic phase was dried,
filtered, concentrated and purified by preparative silica gel plate
(ethyl acetate/petroleum ether: 1/3) to yield a yellow solid
product.
Step 2: Synthesis of methyl
2-((5-cyano-[2,3'-bipyridine]-4'-yl)thio)-2-methylpropanoate
(37-b)
[0225] In a single-necked flask (50 mL), 37-a (108 mg, 0.5 mmol)
and sodium sulfide (117 mg, 1.5 mmol) were dissolved in dimethyl
formamide (10 mL), the mixture was heated to 130.degree. C. and
reacted for 1 hour. After being cooled, the mixture was further
added with anhydrous potassium carbonate (207 mg, 1.5 mmol) and
methyl 2-bromoisobutyrate (272 mg, 1.5 mmol), and was allowed to
further react for 1 hour at 130.degree. C. The reaction solution
was added with ethyl acetate (50 mL), and washed with water (50 mL)
and brine (50 mL). The organic phase was dried, filtered,
concentrated and purified by preparative silica gel plate (ethyl
acetate/petroleum ether: 1/1) to yield a yellow oily substance.
Step 3: Synthesis of
2-((5-cyano-[2,3'-bipyridine]-4'-yl)thio)-2-methylpropanoic acid
(37)
[0226] In a single-necked flask (50 mL), 37-b (60 mg, 0.19 mmol)
and lithium hydroxide (41 mg, 0.97 mmol) were added to
tetrahydrofuran/water (3 mL/1 mL) and reacted at room temperature
for 6 hours. The reaction solution was adjusted to pH=4 with dilute
hydrochloric acid (1 M), added with ethyl acetate (50 mL) and
washed with water (50 mL) and brine (50 mL). The organic phase was
dried, filtered, concentrated and preparatively purified to yield a
white solid product.
[0227] LC-MS (ES, m/z): 300 [M+H].sup.+; H-NMR: (400 MHz,
CD.sub.3OD, ppm): .delta. 9.02 (d, J=2 Hz, 1H), 8.59 (s, 1H), 8.48
(d, J=5.2 Hz, 1H), 8.29-8.26 (m, 1H), 7.90-7.88 (m, 1H), 7.62 (d,
J=5.6 Hz, 1H), 1.50 (s, 6H).
Example 36: Synthesis of Compound 19
##STR00072##
[0229] Compound 19 was synthesized by a method similar to that in
Example 35, except that 6-bromopyridin-3-carbonitrile was replaced
with the corresponding compound in step 1.
[0230] LC-MS (ES, m/z): 314 [M+H].sup.+; H-NMR (400 MHz,
CD.sub.3OD, ppm): .delta. 8.81 (d, J=1.6 Hz, 1H), 8.49 (d, J=6.4
Hz, 1H), 8.31 (s, 1H), 8.18 (d, J=1.2 Hz, 1H), 7.61 (d, J=6.4 Hz,
1H), 2.22 (s, 3H), 1.53 (s, 6H).
Example 37: Synthesis of Compound 38
##STR00073##
[0231] Step 1: Synthesis of methyl
2-(1-(((5-cyano-[2,3'-bipyridine]-4'-yl)thio)methyl)cyclopropyl)propionat-
e (38-a)
[0232] In a single-necked flask (50 mL), 37-a (100 mg, 0.46 mmol)
and methyl 2-(1-(mercaptomethyl)cyclopropyl)propionate (160 mg,
0.92 mmol) were dissolved in dimethyl formamide (2 mL), anhydrous
potassium carbonate (256 mg, 1.84 mmol) was added thereto, and the
reaction was carried out at 130.degree. C. for 0.5 hour. The
reaction solution was added with ethyl acetate (50 mL), and washed
with water (50 mL) and brine (50 mL). The organic phase was dried,
filtered, concentrated and purified by preparative silica gel plate
(ethyl acetate/petroleum ether: 1/1) to yield a white solid.
Step 2: Synthesis of
2-(1-(((5-cyano-2,3'-bipyridin-4'-yl)thio)methyl)
cyclopropyl)acetic acid (38)
[0233] In a single-necked flask (50 mL), 38-a (90 mg, 0.26 mmol)
and lithium hydroxide (41 mg, 0.97 mmol) were added to
tetrahydrofuran/water (3 mL/1 mL) and reacted at 0.degree. C. for 6
hours. The reaction solution was adjusted to pH=4 with dilute
hydrochloric acid (1 M), added with ethyl acetate (50 mL), and
washed with water (50 mL) and brine (50 mL). The organic phase was
dried, filtered, concentrated and preparatively purified to yield a
white solid product.
[0234] LC-MS (ES, m/z): 326 [M+H].sup.+; H-NMR: (400 MHz,
CD.sub.3OD, ppm): 9.11 (d, J=1.6 Hz, 1H), 8.73 (s, 1H), 8.56 (d,
J=6.0 Hz, 1H), 8.38 (dd, J=2.0 Hz, J=8.0 Hz, 1H), 8.02-8.04 (m,
2H), 3.44 (s, 2H), 2.39 (s, 2H), 0.67 (m, 4H).
Example 38: Synthesis of Compound 39
##STR00074##
[0235] Step 1: Synthesis of
4'-(((1-(carboxymethyl)cyclopropyl)methyl)thio)-[2,3'-bipyridine]-5-carbo-
xylic acid (39)
[0236] In a single-necked flask (50 mL), 38-a (40 mg, 0.12 mmol)
and lithium hydroxide (15 mg, 0.36 mmol) were added to
tetrahydrofuran/water (3 mL/1 mL) and reacted for 16 hours. The
reaction solution was adjusted to pH=4 with dilute hydrochloric
acid (1 M), added with ethyl acetate (50 mL), and washed with water
(50 mL) and brine (50 mL). The organic phase was dried, filtered,
concentrated and preparatively purified to yield a white solid
product.
[0237] LC-MS (ES, m/z): 345.0 [M+H].sup.+; H-NMR: (400 MHz,
CD.sub.3OD, ppm): 9.30 (d, J=1.2 Hz, 1H), 8.72 (s, 1H), 8.55 (m,
2H), 8.04 (d, J=6.4 Hz, 1H), 7.95 (d, J=8.4 Hz, 1H), 3.45 (s, 2H),
2.39 (s, 2H), 0.67 (m, 4H).
Example 39: Synthesis of Compound 32
##STR00075##
[0238] Step 1: Synthesis of ethyl 3-(3-bromopyridin-4-yl)propanoate
(32-a)
[0239] In a single-necked flask (100 mL), under the protection of
nitrogen, 3-bromo-4-methylpyridine (500 mg, 2.9 mmol) was dissolved
in the tetrahydrofuran (10 mL), the solution was then cooled to
-78.degree. C., and added with homemade lithium diisopropylamide
(LDA) (3.5 mL, 3.5 mmol), after reacted for 1 hour, ethyl
2-bromoacetate (1.22 g, 7.3 mmol) was further added dropwise
thereto and reacted for another 2 hours. The reaction was quenched
with saturated sodium bicarbonate solution, the reaction solution
was added with ethyl acetate (50 mL), and washed with water (50 mL)
and brine (50 mL). The organic phase was dried, filtered,
concentrated and purified by preparative silica gel plate
(petroleum ether/ethyl acetate: 2/1) to yield a yellow oily
substance.
Step 2: Synthesis of ethyl
3-(3-(4-cyanonaphthalen-1-yl)pyridin-4-yl) propanoate (32-b)
[0240] 32-a (100 mg, 0.39 mmol), the aqueous solution of sodium
carbonate (0.8 mL, 1.6 mmol, 2 M),
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-cyano-naphthalene
(108 mg, 0.39 mmol), and [1,1'-bis(diphenylphosphino)
ferrocene]dichloropalladium (29 mg, 0.04 mmol) were added to
dimethyl formamide (3 mL) in a single-necked flask (50 mL), purged
with nitrogen 3 times, and then the mixture was heated to
130.degree. C. and reacted for 5 hours. The reaction solution was
added with ethyl acetate (50 mL), and washed with water (50 mL) and
brine (50 mL). The organic phase was dried, filtered, concentrated
and purified by preparative silica gel plate (petroleum ether/ethyl
acetate: 1/1) to yield a light yellow solid.
Step 3: Synthesis of 3-(3-(4-cyanonaphthalen-1-yl)pyridin-4-yl)
propanoic acid (32)
[0241] In a single-necked flask (50 mL), 32-b (30 mg, 0.1 mmol) and
lithium hydroxide (41 mg, 0.97 mmol) were added to
tetrahydrofuran/water (3 mL/1 mL), and the mixture was reacted at
room temperature for 16 hours. The reaction solution was adjusted
to pH=4 with dilute hydrochloric acid (1M), added with ethyl
acetate (50 mL), and then washed with water (50 mL) and brine (50
mL). The organic phase was dried, filtered, concentrated, and
preparatively purified to yield a white solid product.
[0242] LC-MS (ES, m/z): 301 [M-H].sup.-; H-NMR: (400 MHz,
CD.sub.3OD, ppm): .delta. 8.60 (d, J=5.2 Hz, 1H), 8.36 (s, 1H),
8.32-8.30 (m, 1H), 8.15-8.13 (m, 1H), 7.83-7.79 (m, 1H), 7.69-7.65
(m, 1H), 7.63-7.59 (m, 2H), 7.51-7.48 (m, 1H), 2.73-2.56 (m, 2H),
2.45-2.40 (m, 2H).
Example 40: Synthesis of Compound 40
##STR00076##
[0243] Step 1: Synthesis of ethyl 2-(acetylthio)acetate (40-a)
[0244] In a three-necked flask (250 mL), the mixture of ethyl
2-bromoacetate (4.17 g, 0.025 mol) and potassium thioacetate (5.7
g, 0.05 mol) was dissolved in dimethyl formamide (100 mL), and the
resulting solution was stirred at room temperature overnight. The
reaction solution was added with water and ethyl acetate, organic
layer was washed with saturated brine and dried over sodium
sulfate, the solvent was removed by reduced pressure evaporation,
thereby a brown oily substance was obtained and directly subject to
the next step for reaction.
Step 2: Synthesis of ethyl
2-((3-(4-cyanonaphthalen-1-yl)pyridin-4-yl) thio)acetate (40-b)
[0245] In a single-necked flask (100 mL), 40-a (1.46 g, 9 mmol),
anhydrous potassium carbonate (1.24 g, 9 mmol) and 1-b (0.795 g, 3
mmol) were added to dimethyl formamide (20 mL), the mixture was
stirred at room temperature for 1 hour, then heated to 130.degree.
C., and stirred with heating for 1 hour. After being cooled to room
temperature, the reaction solution was added with water and ethyl
acetate, organic layer was washed with saturated brine, dried over
sodium sulfate, then the solvent was removed by reduced pressure
evaporation, to give a crude brown oil, which was purified by
column chromatography (petroleum ether/ethyl acetate=1:1) to yield
an oil.
[0246] H-NMR: (400 MHz, CDCl.sub.3, ppm): .delta. 8.60 (d, J=5.6
Hz, 1H), 8.35-8.33 (m, 2H), 7.99 (d, J=7.2 Hz, 1H), 7.75-7.72 (m,
1H), 7.58-7.55 (M, 2H), 7.49 (d, J=7.2 Hz, 1H), 7.36-7.34 (m, 1H),
4.20-4.16 (m, 2H), 3.63 (s, 2H), 1.26-1.24 (m, 3H).
Step 3: Synthesis of
di(methyl-d3)-2-((3-(4-cyanonaphthalen-1-yl)pyridin-4-yl)thio)acetic
acid (40)
[0247] In a three-necked flask (50 mL), 40-b (110 mg, 0.3 mmol)
dissolved in tetrahydrofuran (1 mL) was slowly added dropwise into
a suspension of sodium hydride (60%, 28 mg, 0.69 mmol) in dimethyl
formamide (1 mL) at 0.degree. C., after being stirred for 10 min,
the mixture was further added dropwise with a solution of
iodomethane-d3 (136 mg, 0.94 mmol) in dimethyl formamide (1 mL) at
0.degree. C., then the mixture was stirred at room temperature
overnight. After being quenched with water, the reaction solution
was adjusted to pH=4 with 1 N of hydrochloric acid, then the
solvent was removed by reduced pressure evaporation, and the
remaining oil was purified by preparative HPLC to yield a light
yellow solid.
[0248] LC-MS (ES, m/z): 355 [M+H].sup.+; H-NMR: (400 MHz,
CD.sub.3OD, ppm): .delta. 8.52-8.50 (m, 1H), 8.30-8.27 (m, 2H),
8.12-8.09 (m, 1H), 7.82-7.77 (m, 1H), 7.68-7.60 (m, 2H), 7.55-7.48
(m, 2H).
Example 41: Synthesis of Compound 41
##STR00077##
[0249] Step 1: Synthesis of ethyl
2-((3-(4-cyanonaphthalen-1-yl)pyridin-4-yl) thio)propanoate
(41-a)
[0250] In a single-necked flask (50 mL), 1-b (264 mg, 1 mmol) and
sodium sulfide (234 mg, 3 mmol) were added to dimethyl formamide
(10 mL), the mixture was heated to 130.degree. C. and reacted for 1
hour. After being cooled, the mixture was further added with
hydrous potassium carbonate (414 mg, 3 mmol) and ethyl
2-bromoacetate (716 mg, 4 mmol), then heated to 130.degree. C. and
was allowed to further react for 1 hour. After being cooled, the
reaction solution was added with ethyl acetate (50 mL), and washed
with water (50 mL) and brine (50 mL). The organic phase was dried,
filtered, concentrated and purified by preparative silica gel plate
(ethyl acetate/petroleum ether: 1/2) to yield a white solid
product.
[0251] H-NMR: (400 MHz, d6-DMSO, ppm): .delta. 8.61 (dd, J=2.0 Hz,
J=5.2 Hz, 1H), 8.34 (d, J=5.6 Hz, 1H), 8.33-8.25 (m, 1H), 8.22 (d,
J=8.4 Hz, 1H), 7.85-7.84 (m, 1H), 7.66-7.63 (m, 1H), 7.63 (d, J=6.4
Hz, 1H), 7.58 (dd, J=3.2 Hz, J=7.6 Hz, 1H), 7.44-7.41 (m, 1H),
4.44-4.39 (m, 1H), 4.07-4.02 (m, 2H), 1.32-1.30 (m, 3H), 1.10-1.06
(m, 3H).
Step 2: Synthesis of ethyl
2-methyl-d3-2-((3-(4-cyanonaphthalen-1-yl)
pyridin-4-yl)thio)propanoate (41-b)
[0252] In a three-necked flask (100 mL), under the protection of
nitrogen, sodium hydride (16 mg, in oil (60%), 0.4 mmol) was added
to dimethyl formamide (5 mL), the mixture was then cooled to
0.degree. C., and a solution of 41-a (120 mg, 0.33 mmol) in
tetrahydrofuran (2.5 mL) was added thereto. The mixture was reacted
at 0.degree. C. for 0.5 hour, the reaction solution was further
added dropwise with a solution of iodomethane-d3 (58 mg, 0.4 mmol)
in dimethyl formamide (1.5 mL), then heated to room temperature and
reacted for another 16 hours. The reaction solution was added with
ethyl acetate (50 mL), and washed with water (20 mL) and brine (10
mL). The organic phase was dried, filtered, concentrated and
purified by preparative silica gel plate (ethyl acetate/petroleum
ether: 1/2) to yield a white solid product.
[0253] H-NMR: (400 MHz, d6-DMSO, ppm): .delta. 8.64 (d, J=5.2 Hz,
1H), 8.41 (s, 1H), 8.28 (d, J=7.2 Hz, 1H), 8.22 (d, J=8.4 Hz, 1H),
7.83 (t, J=7.6 Hz, 1H), 7.69 (t, J=7.6 Hz, 1H), 7.57 (d, J=7.2 Hz,
1H), 7.42-7.38 (m, 2H), 4.08 (q, J=7.2 Hz, 2H), 1.37 (d, J=3.2 Hz,
3H), 1.10 (t, J=7.2 Hz, 3H).
Step 3: Synthesis of
2-methyl-d3-2-((3-(4-cyanonaphthalen-1-yl)pyridin-4-yl)thio)
propanoic acid (41)
[0254] In a single-necked flask (50 mL), 41-b (60 mg, 0.16 mmol)
and lithium hydroxide (41 mg, 0.97 mmol) were added to
tetrahydrofuran/water (3 mL/1 mL) and reacted at room temperature
for 12 hours. The reaction solution was adjusted to pH=4 with
dilute hydrochloric acid (1 M), added with ethyl acetate (30 mL),
and washed with brine (20 mL). The organic phase was dried,
filtered, concentrated and preparatively purified to yield a white
solid product.
[0255] LC-MS (ES, m/z): 352 [M-H].sup.-; H-NMR: (400 MHz, d6-DMSO,
ppm): .delta. 13.18 (br, 1H), 8.61 (d, J=5.2 Hz, 1H), 8.36 (s, 1H),
8.26 (d, J=7.2 Hz, 1H), 8.21 (d, J=8.4 Hz, 1H), 7.83 (t, J=7.6 Hz,
1H), 7.66 (t, J=7.6 Hz, 1H), 7.57 (d, J=7.2 Hz, 1H), 77.51 (d,
J=5.2 Hz, 1H), 7.41 (d, J=8.4 Hz, 1H), 1.37 (d, J=10.8 Hz, 3H).
Experimental Example 1: Evaluation of Bioactivity of Chemicals for
Inhibiting Absorption of Uric Acid Using URAT1 Cell Model
[0256] Human kidney embryonic cells HEK-293T were grown in a petri
dish (diameter=10 cm) containing DMEM and 10% of bovine fetal serum
culture solution, and incubated in an 5% of carbon
dioxide-containing incubator at 37.degree. C. Plasmids carrying
human URAT1 were transfected to HEK-293T cells using TranslT-293
(Mirus Bio LLC). After 72 hours, the petri dish containing HEK-293T
cells transfected with URAT1 was removed from the incubator and the
cells were inoculated on Poly-D-Lysine Coated 96-well Plates at a
density of 60,000 cells per well. After the cells on the 96-well
plates were grown overnight (at least 12 hours) in an incubator at
37 degrees, these cells were gently rinsed 3 times with warm and no
chloride ions-containing HBSS buffer (125 mM sodium gluconate, 4.8
mM potassium gluconate, 1.3 mM calcium gluconate, 1.2 mM
monopotassium phosphate, 1.2 mM magnesium sulfate, 5.6 mM glucose,
25 mM HEPES, pH 7.4). 50 microliter of HBSS buffer (not containing
chloride ions) containing 0.2 microcurie of .sup.14C-uric acid and
compounds of the present application or benzbromarone, and vector
was added in each well, then the cell plates were put back to the
incubator at 37 degrees. After 5 min, the buffer was removed from
cell wells, added with 100 microliter of ice-cold and no chloride
ions-containing HBSS buffer to gently rinse cells within wells so
as to stop them from absorbing .sup.14C-uric acid, the rinsing was
repeated 3 times in the same manner. 150 microliter of cell lysate
(100 mM of NaOH) was added in each well. Cell plate was placed on a
vibrating plate and vibrated for 10 min at a speed of 600 rpm such
that the cells were completely lysed. The cell plate was put in a
centrifuge and spun for 5 min at a speed of 1000 rpm, then 45
microliter of supernatant was sucked out from each well and
transferred to 96-well plate (Isoplate-96 Microplate from
PerkinElmer). In the new 96-well plate, 150 microliter of Ultima
Gold XR scintillation solution was added in each well. The 96-well
plate was vibrated for 10 min at a speed of 600 rpm on a vibrating
plate. Finally, the 96-well plate was put in a MicroBeta Trilux
Counter from PerkinElmer and was read, then IC.sub.50 values were
calculated, and the results are shown in Table 1 below. Wherein
[0257] I represents that IC.sub.50 value is in the range of less
than or equal to 100 nM; [0258] II represents that IC.sub.50 value
is in the range of less than or equal to 1000 nM and more than 100
nM; and [0259] III represents that IC.sub.50 value is more than
1000 nM.
TABLE-US-00001 [0259] TABLE 1 Compound URAT1 IC.sub.50 No. Activity
level 1 I 2 II 3 I 4 I 5 II 6 III 7 II 8 III 9 III 10 III 11 I 12
II 13 II 14 II 15 II 16 I 17 I 18 I 19 III 20 I 21 I 22 II 23 III
24 III 25 III 26 III 27 III 28 III 29 III 30 III 31 III 32 III 33
II 34 II 35 II 36 III 37 III 38 III 39 III 40 I 41 I benzbromarone
II
[0260] From the experimental data listed in the above Table 1, it
can be seen that, compared with existing compound benzbromarone,
the compounds of the present invention have lower or similar
IC.sub.50 values, thereby it is demonstrated that the compounds of
the present invention have higher activities of inhibiting the
reabsorption of uric acid, and can be used as novel and efficient
drugs for reducing blood uric acid.
[0261] The examples and embodiments disclosed herein are merely for
the illustrative purpose, and various amendments and modifications
made by a person skilled in the art will be included in the spirit
and scope of the present application and are within the scope of
the appended claims.
* * * * *