U.S. patent application number 16/305644 was filed with the patent office on 2020-04-23 for pharmaceutically acceptable salt as renal outer medullary potassium channel inhibitor.
The applicant listed for this patent is Jiangsu Hengrui Medicine Co., Ltd.. Invention is credited to Jun FENG, Yun LU, Qiyun SHAO, Guaili WU.
Application Number | 20200123135 16/305644 |
Document ID | / |
Family ID | 60577585 |
Filed Date | 2020-04-23 |
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United States Patent
Application |
20200123135 |
Kind Code |
A1 |
LU; Yun ; et al. |
April 23, 2020 |
PHARMACEUTICALLY ACCEPTABLE SALT AS RENAL OUTER MEDULLARY POTASSIUM
CHANNEL INHIBITOR
Abstract
A pharmaceutically acceptable salt as a renal outer medullary
potassium channel (ROMK) inhibitor, specifically an L-tartrate and
a crystal form I and crystal form II thereof, is described. The
pharmaceutically acceptable salt of the ROMK inhibitor,
specifically the L-tartrate, has improved bioavailability and
stability.
Inventors: |
LU; Yun; (Lianyungang,
Jiangsu, CN) ; SHAO; Qiyun; (Lianyungang, Jiangsu,
CN) ; WU; Guaili; (Lianyungang, Jiangsu, CN) ;
FENG; Jun; (Lianyungang, Jiangsu, CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Jiangsu Hengrui Medicine Co., Ltd. |
Lianyungang, Jiangsu |
|
CN |
|
|
Family ID: |
60577585 |
Appl. No.: |
16/305644 |
Filed: |
June 6, 2017 |
PCT Filed: |
June 6, 2017 |
PCT NO: |
PCT/CN2017/087326 |
371 Date: |
November 29, 2018 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 9/04 20180101; A61P
9/12 20180101; C07D 405/14 20130101; A61K 31/4545 20130101 |
International
Class: |
C07D 405/14 20060101
C07D405/14; A61P 9/12 20060101 A61P009/12; A61P 9/04 20060101
A61P009/04 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 7, 2016 |
CN |
201610398233.0 |
Claims
1. A pharmaceutically acceptable salt of
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide, selected from the
group consisting of tartrate, hydrochloride, sulfate,
methanesulfonate, phosphate, citrate, maleate, fumarate, malate,
benzenesulfonate, p-toluenesulfonate and hydrobromide.
2. The salt according to claim 1, wherein the salt is L-tartrate or
malate.
3. A compound of formula (I), ##STR00006##
4. A method for preparing the compound of formula (I) according to
claim 3, comprising reacting
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide with L-tartaric
acid.
5. Crystal form I of the compound of formula (I) according to claim
3, characterized by an X-ray powder diffraction spectrum comprising
diffraction peaks at diffraction angles 2.theta..+-.0.2 of 3.49,
10.22, 12.27, 13.69, 15.46, 16.98, 18.04, 19.45, 23.95, and
29.44.
6. A method for preparing the crystal form I of the compound of
formula (I) according to claim 5, comprising: 1) adding any crystal
form or amorphous form of the compound of formula (I), or adding
L-tartaric acid and
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofura-
n-5-yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide to a solvent to
obtain a mixture and then precipitating a crystal from the mixture,
wherein the solvent is selected from the group consisting of an
alcohol having 3 or less carbon atoms, and a mixed solvent of the
alcohol and water; and 2) filtering the crystal, then washing and
drying the crystal.
7. The method according to claim 6, wherein the solvent in step 1)
is selected from the group consisting of methanol, ethanol, and
isopropanol or an aqueous solution thereof.
8. Crystal form II of the compound of formula (I) according to
claim 3, characterized by an X-ray powder diffraction spectrum
comprising diffraction peaks at diffraction angles 2.theta..+-.0.2
of 3.88, 13.00, 17.25, 25.42, and 27.79.
9. A method for preparing the crystal form II according to claim 8,
comprising the: 1) adding any crystal form or amorphous form of the
compound of formula (I), or adding L-tartaric acid and
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide to a solvent to
obtain a mixture and then precipitating a crystal from the mixture,
wherein the solvent is a mixed solvent of
isopropanol/tetrahydrofuran/water; and 2) filtering the crystal,
then washing and drying the crystal.
10. The salt according to claim 2, wherein a chemical ratio of
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide to malic acid is
1:1, and the salt is a compound of formula (I'); ##STR00007##
11. The salt according to claim 2, wherein a chemical ratio of
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide to malic acid is
1:0.5, and the salt is a compound of formula (I'');
##STR00008##
12. Crystal form 1 of the compound of formula (I') according to
claim 10, characterized by an X-ray powder diffraction spectrum
comprising diffraction peaks at diffraction angles 2.theta..+-.0.2
of 6.85, 9.76, 11.81, 12.48, 13.52, 14.23, 15.29, 17.55, 17.89,
19.65, 20.25, 22.68(3.92), 23.50, 24.27, 27.83, and 28.59.
13. A method for preparing the crystal form 1 according to claim
12, wherein the method is selected from the group consisting of:
(1) reacting
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide with malic acid in a
solvent or a mixed solvent, followed by stirring, precipitation,
filtration and drying to obtain the crystal form 1; wherein the
solvent is an alcohol solvent, and the mixed solvent is a mixed
solvent of an alcohol and water, and (2) adding the compound of
formula (I') to a solvent, followed by pulping, filtration and
drying to obtain the crystal form 1; wherein the solvent is a mixed
solvent of an alcohol and water.
14. Crystal form 2 of the compound of formula (I') according to
claim 10, characterized by an X-ray powder diffraction spectrum
comprising diffraction peaks at diffraction angles 2.theta..+-.0.2
of 9.34, 10.61(8.33), 12.24, 12.78, 13.83, 14.26, 15.52, 17.83,
18.83, 19.86, 20.55, 21.72, 22.31, 22.62, 23.42, 24.78, 25.03,
25.27, 25.78, 26.02, 26.76, 27.99, 30.72, 31.19, 34.41, 36.68,
39.87, and 41.25.
15. A method for preparing the crystal form 2 according to claim
14, wherein the method is selected from the group consisting of:
(1) reacting
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide with malic acid in a
solvent or a mixed solvent, followed by stirring, precipitation,
filtration and drying to obtain the crystal form 2; wherein the
solvent is selected from the group consisting of an alcohol solvent
and a nitrile solvent, and the mixed solvent is selected from the
group consisting of a mixed solvent of a nitrile and water and a
mixed solvent of a nitrile and an ether; and (2) adding the
compound of formula (I') to a solvent or a mixed solvent, followed
by pulping, filtration and drying to obtain the crystal form 2;
wherein the solvent is selected from the group consisting of an
alcohol solvent, a nitrile solvent and a ketone solvent, and the
mixed solvent is selected from the group consisting of a mixed
solvent of a nitrile and water and a mixed solvent of a nitrile and
an ether.
16. Crystal form 3 of the compound of formula (I') according to
claim 10, characterized by an X-ray powder diffraction spectrum
comprising diffraction peaks at diffraction angles 2.theta..+-.0.2
of 9.34, 10.63, 12.79, 14.22, 15.50, 18.86, 19.93, and 25.16.
17. A method for preparing the crystal form 3 according to claim
16, wherein the method is selected from the group consisting of:
(1) reacting
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide with malic acid in a
solvent or a mixed solvent, followed by stirring, precipitation,
filtration and drying to obtain the crystal form 3; wherein the
solvent is selected from the group consisting of an alcohol solvent
and an ketone solvent, and the mixed solvent is selected from the
group consisting of a mixed solvent of an alcohol and water; and
(2) adding the compound of formula (I') to a solvent or a mixed
solvent, followed by pulping, filtration and drying to obtain the
crystal form 3; wherein the solvent is selected from the group
consisting of an alcohol solvent and a ketone solvent, and the
mixed solvent is a mixed solvent of an alcohol and water.
18. Crystal form A of the compound of formula (I'') according to
claim 11, characterized by an X-ray powder diffraction spectrum is
comprising diffraction peaks at diffraction angles 2.theta..+-.0.2
of 9.95, 12.12, 12.68, 13.56, 14.73, 15.32, 16.20, 17.27, 18.74,
20.00, 21.57, 22.82, 23.22, 24.35, 24.63, 25.42, 27.13, 28.15,
30.01, 30.30, 32.92, and 34.97.
19. A method for preparing the crystal form A according to claim
18, wherein the method is selected from the group consisting of:
(1) reacting
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide with malic acid in a
solvent or a mixed solvent, followed by stirring, precipitation,
filtration and drying to obtain the crystal form A; wherein the
solvent is an alcohol solvent, and the mixed solvent is a mixed
solvent of an alcohol and water; and (2) adding the compound of
formula (I'') to a solvent or a mixed solvent, followed by pulping,
filtration and drying to obtain the crystal form A; wherein the
solvent is an alcohol solvent, and the mixed solvent is a mixed
solvent of an alcohol and water.
20. A pharmaceutical composition, comprising the salt according to
claim 1 and a pharmaceutically acceptable carrier.
21.-22. (canceled)
23. A method for the treatment or prevention of a disease related
to renal outer medullary potassium channel (ROMK) inhibition,
comprising administering to a subject in need thereof a
therapeutically effective amount of the pharmaceutical composition
according to claim 20.
24. The method according to claim 23, wherein the disease is
hypertension or heart failure.
Description
CROSS REFERENCE TO RELATED APPLICATION
[0001] This application is a Section 371 of International
Application No. PCT/CN2015/087326, filed Jun. 6, 2017, which was
published in the Chinese language on Dec. 14, 2017 under
International Publication No. WO 2017/211271 A1, which claims
priority under 35 U.S.C. .sctn. 119(b) to Chinese Application No.
201610398233.0, filed Jun. 7, 2016, the disclosures of which are
incorporated herein by reference in their entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to a pharmaceutically
acceptable salt of
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-
-5-yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide.
BACKGROUND OF THE INVENTION
[0003] Increased renal salt reabsorption function can cause a risk
of hypertension. On the contrary, inhibition of renal reabsorption
function can promote the excretion of urine, which results in
diuretic and antihypertensive effects. Common diuretics are
thiazide diuretics, which are first-line antihypertensive drugs in
USA that primarily act on Na.sup.+--Cl.sup.- transporters. The Loop
diuretics are more effective for patients with impaired renal
function, and they play a role through Na.sup.+--K.sup.+-2Cl.sup.-
transport proteins. However, both drugs can cause hypokalemia
(symptoms: weakness, fatigue, muscle cramps, constipation, and
heart rhythm problems, such as arrhythmia), which increases the
risk of morbidity and mortality of cardiovascular diseases.
[0004] Renal Outer Medullary Potassium channel (ROMK) is also known
as the inward-rectifying potassium channel 1.1 (Kir1.1). The ROMK
ion channel, cooperating with the Na.sup.+--K.sup.+-2Cl.sup.-
co-transport protein NKCC2 (responsible for NaCl transport) through
the apical membrane conductance of the renal thick ascending limb
(TAL), can regulate the reabsorption of Na.sup.+. The ROMK was
found to be directly associated with the renal secretory channel.
When the ROMK gene is knocked out in mice, there is a loss of TAL
and CCD 35-pS ion channels as well as a loss of the other K.sup.+
channels in TAL. Batter syndrome is an autosomal recessive disease
characterized by massive loss of salt in the kidneys, hypokalemia,
and low blood pressure. Batter syndrome is mainly caused by
mutations in the ROMK or Na.sup.+--K.sup.+-2Cl.sup.- co-transport
proteins. The difference is that the hypokalemia of the batter
syndrome caused by the mutation of ROMK is much milder compared to
that caused by the mutation of Na.sup.+--K.sup.+-2Cl.sup.-
co-transport proteins. In summary, inhibition of ROMK function can
effectively inhibit the salt reabsorption function of
Na.sup.+--K.sup.+-2Cl.sup.- co-transport proteins and promote the
excretion of urine, thereby resulting in diuretic and
antihypertensive effects, without causing hypokalemia.
[0005] WO2016091042, which was previously filed by the applicant
and is incorporated herein by reference in its entirety, discloses
a series of renal outer medullary potassium channel (ROMK)
inhibitors, comprising a compound represented by the following
formula, with the chemical name of
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide. Compared to other
ROMK inhibitors, this compound is added with a polar group, which
reduces C log P, enhances the hERG selectivity and improves safety,
while maintaining the ROMK inhibitor activity.
##STR00001##
[0006] However, the compound is present in the form of a free base
with poor bioavailability and stability. Therefore, it is necessary
to find a suitable form for the development to solve the problem of
low bioavailability and low stability. Moreover, the crystal
structure of the pharmaceutically active ingredient often affects
the chemical stability of the drug. Different crystallization
conditions and storage conditions can lead to changes in the
crystal structure of the compound, and sometimes accompanying
production of other crystal forms. In general, an amorphous drug
product does not have a regular crystal structure, and often has
other defects, such as poor product stability, finer
crystallization, difficult filtration, easy agglomeration, and poor
liquidity. Therefore, it is necessary to improve the various
properties of the above compound. There is a need to find a new
crystal form with high purity and good chemical stability.
SUMMARY OF THE INVENTION
[0007] The present invention provides a pharmaceutically acceptable
salt, in particular an acid addition salt of
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide. The
bioavailability and stability can be greatly improved after salt
formation.
[0008] In a preferred embodiment, the salt can be selected from the
group consisting of tartrate, hydrochloride, sulfate,
methanesulfonate, phosphate, citrate, maleate, fumarate, malate,
benzenesulfonate, p-toluenesulfonate and hydrobromide, and most
preferably L-tartrate and malate. The L-tartrate of
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide, i.e. the compound
of the following formula (I), can be obtained by reacting
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide with L-tartaric
acid. The hydrochloride, sulfate, methanesulfonate, phosphate,
citrate, maleate, fumarate, malate, benzenesulfonate,
p-toluenesulfonate and hydrobromide of
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-
-5-yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide can be prepared
by using a similar method. Since the solubility of the compound of
formula (II) is low, in order to further increase its solubility,
the applicant has carried out a salt formation study on the
compound of formula (II), wherein the salt formation of the
compound of formula (II) with tartaric acid, hydrochloric acid,
sulfuric acid, methanesulfonic acid, phosphoric acid, citric acid,
maleic acid, fumaric acid, malic acid, benzenesulfonic acid,
p-toluenesulfonic acid, hydrobromic acid or lactic acid has been
investigated. The solubility of the compound has been greatly
improved after salt formation. Whether the salt is easy to form
crystals etc. has also been investigated. It has an important
research significance in the treatment of ROMK related diseases.
During the salt formation process, the applicant has found that the
reaction between lactic acid and the compound of formula (II) does
not result in a salt, the ratio of fumaric acid to the compound of
formula (II) and the crystal form are difficult to control when
forming a fumarate, the sulfate and citrate are obviously
hygroscopic, the crystal forms of phosphate are complicated and
exist simultaneously, which leads to difficulty in separation, and
there are many types of the crystal forms of maleate.
##STR00002##
[0009] A series of crystal products of the compound of formula (I)
have been obtained under different crystallization conditions, and
X-ray diffraction and differential scanning calorimetry (DSC)
measurements have been conducted on the obtained crystal products.
It was found that a stable crystal form, which is referred to as
crystal form I, can be obtained under crystallization conditions of
the present invention. The DSC spectrum of crystal form I of the
present application shows a melting endothermic peak at about
218.42.degree. C. The X-ray powder diffraction spectrum, which is
obtained by using Cu-K.alpha. radiation and represented by
diffraction angle 2.theta. angle, is shown in FIG. 1, in which
there are characteristic peaks at diffraction angle 2.theta. angles
of 3.49, 10.22, 12.27, 13.69, 15.46, 16.98, 18.04, 19.45, 23.95 and
29.44, wherein the error range of 2.theta. angle of each
characteristic peak is .+-.0.2.
[0010] The present invention also provides a method for preparing
crystal form I of the tartrate of
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide, comprising the
following steps of:
[0011] 1) adding L-tartaric acid to an appropriate amount of
solvent and dissolving it under heating, then adding any crystal
form or amorphous form of
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzo-
furan-5-yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide to form a
salt; wherein the solvent is selected from the group consisting of
an alcohol having 3 or less carbon atoms, and a mixed solvent of
the alcohol and water; and
[0012] 2) filtering the crystal, then washing and drying it.
[0013] In a preferred embodiment, the solvent in step 1) is
methanol, ethanol, isopropanol or an aqueous solution of the
alcohol; wherein the single solvent is preferably methanol.
[0014] In an embodiment of the present invention, the preferred
mixed organic solvent is a mixed solvent of ethanol/water, and the
ratio of the two is not particularly limited. In a preferred
embodiment of the present invention, the volume ratio of the two is
9:1.
[0015] Crystal form II of the compound of formula (I) can be
obtained under another crystallization condition, wherein the X-ray
powder diffraction spectrum thereof is obtained by using
Cu-K.alpha. radiation and represented by diffraction angle 2.theta.
angle, in which there are characteristic peaks at diffraction angle
2.theta. angles of 3.88, 13.00, 17.25, 25.42 and 27.79, wherein the
error range of 2.theta. angle of each characteristic peak is
.+-.0.2.
[0016] The method for preparing the crystal form II can comprise
the following steps of:
[0017] 1) adding any crystal form or amorphous form of the compound
of formula (I), or adding L-tartaric acid and
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide to a solvent to
dissolve and precipitate a crystal, wherein the solvent is a mixed
solvent of isopropanol/tetrahydrofuran/water; and
[0018] 2) filtering the crystal, then washing and drying it.
[0019] The recrystallization method is not particularly limited,
and can be carried out by a conventional recrystallization process.
For example, the material, i.e., the compound of formula (I), can
be dissolved in an organic solvent under heating, and then the
solution is cooled slowly to precipitate a crystal. After the
completion of crystallization, the desired crystal can be obtained
via filtering and drying. In particular, the crystal obtained by
filtration is usually dried in a vacuum under reduced pressure at a
heating condition of about 30 to 100.degree. C., preferably 40 to
60.degree. C., to remove the recrystallization solvent.
[0020] The present invention also provides a malate, characterized
in that the chemical ratio of
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide to malic acid is
1:1, and the specific structure thereof is shown in the formula
(I'):
##STR00003##
[0021] The present invention further provides a malate, wherein the
chemical ratio of
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide to malic acid is
1:0.5, and the specific structure thereof is shown in formula
(I''):
##STR00004##
[0022] Preferably, the crystal form 1 of the compound of formula
(I') is characterized in that the X-ray powder diffraction spectrum
thereof is obtained by using Cu-K.alpha. radiation and represented
by diffraction angle 2.theta. angle, in which there are
characteristic peaks at diffraction angle 2.theta. angles of 6.85,
9.76, 11.81, 12.48, 13.52, 14.23, 15.29, 17.55, 17.89, 19.65,
20.25, 22.68(3.92), 23.50, 24.27, 27.83 and 28.59, wherein the
error range of 20 angle of each characteristic peak is .+-.0.2.
[0023] The present invention also provides a method for preparing
the crystal form 1 of the compound of formula (I'), wherein the
method is selected from the following preparation methods:
[0024] (1) reacting the free form of the compound of formula (I')
with malic acid in a solvent or a mixed solvent, followed by
stirring, precipitation, filtration and drying to obtain the
desired crystal form 1; wherein the solvent is an alcohol solvent,
the mixed solvent is a mixed solvent of an alcohol and water, the
alcohol solvent is preferably methanol, and the mixed solvent of an
alcohol and water is preferably a mixed solvent of methanol and
water; and
[0025] (2) adding the compound of formula (I') to a solvent,
followed by pulping, filtration and drying to obtain the desired
crystal form 1; wherein the solvent is a mixed solvent of an
alcohol and water, the alcohol solvent is preferably methanol,
ethanol or isopropanol, and the mixed solvent of an alcohol and
water is preferably a mixed solvent of methanol and water or a
mixed solvent of ethanol and water.
[0026] Preferably, the crystal form 2 of the compound of formula
(I') is characterized in that the X-ray powder diffraction spectrum
thereof is obtained by using Cu-K.alpha. radiation and represented
by diffraction angle 2.theta. angle, in which there are
characteristic peaks at diffraction angle 2.theta. angles of 9.34,
10.61(8.33), 12.24, 12.78, 13.83, 14.26, 15.52, 17.83, 18.83,
19.86, 20.55, 21.72, 22.31, 22.62, 23.42, 24.78, 25.03, 25.27,
25.78, 26.02, 26.76, 27.99, 30.72, 31.19, 34.41, 36.68, 39.87 and
41.25, wherein the error range of 2.theta. angle of each
characteristic peak is .+-.0.2.
[0027] The present invention also provides a method for preparing
the crystal form 2 of the compound of formula (I'), characterized
in that the method is selected from the following preparation
methods:
[0028] (1) reacting the free form of the compound of formula (I')
with malic acid in a solvent or a mixed solvent, followed by
stirring, precipitation, filtration and drying to obtain the
desired crystal form 2; wherein the solvent is selected from the
group consisting of an alcohol solvent and a nitrile solvent, the
mixed solvent is selected from the group consisting of a mixed
solvent of a nitrile and water and a mixed solvent of a nitrile and
an ether, preferably, the alcohol solvent is methanol or ethanol,
the nitrile solvent is acetonitrile, and the ether solvent is
tetrahydrofuran; and
[0029] (2) adding the compound of formula (I') to a solvent or a
mixed solvent, followed by pulping, filtration and drying to obtain
the desired crystal form 2; wherein the solvent is selected from
the group consisting of an alcohol solvent, a nitrile solvent and a
ketone solvent, preferably the solvent is selected from the group
consisting of an alcohol solvent and a nitrile solvent, the mixed
solvent is selected from the group consisting of a mixed solvent of
a nitrile and water and a mixed solvent of a nitrile and an ether,
preferably, the alcohol solvent is methanol or ethanol, the nitrile
solvent is acetonitrile, and the ether solvent is
tetrahydrofuran.
[0030] Preferably, the crystal form 3 of the compound of formula
(I') is characterized in that the X-ray powder diffraction spectrum
thereof is obtained by using Cu-K.alpha. radiation and represented
by diffraction angle 2.theta. angle, in which there are
characteristic peaks at diffraction angle 2.theta. angles of 9.34,
10.63, 12.79, 14.22, 15.50, 18.86, 19.93 and 25.16, wherein the
error range of 2.theta. angle of each characteristic peak is
.+-.0.2.
[0031] The present invention also provides a method for preparing
the crystal form 3 of the compound of formula (I'), characterized
in that the method is selected from the following preparation
methods:
[0032] (1) reacting the free form of the compound of formula (I')
with malic acid in a solvent or a mixed solvent, followed by
stirring, precipitation, filtration and drying to obtain the
desired crystal form 3; wherein the solvent is selected from the
group consisting of an alcohol solvent and a ketone solvent, the
mixed solvent is selected from the group consisting of a mixed
solvent of an alcohol and water, the alcohol solvent is preferably
isopropanol, the ketone solvent is acetone, and the mixed solvent
of an alcohol and water is a mixed solvent of isopropanol and
water; and
[0033] (2) adding the compound of formula (I') to a solvent or a
mixed solvent, followed by pulping, filtration and drying to obtain
the desired crystal form 3; wherein the solvent is selected from
the group consisting of an alcohol solvent and a ketone solvent,
the mixed solvent is a mixed solvent of an alcohol and water, the
alcohol solvent is preferably isopropanol, the ketone solvent is
acetone, and the mixed solvent of an alcohol and water is
preferably a mixed solvent of isopropanol and water.
[0034] Preferably, the crystal form A of the compound of formula
(I'') is characterized in that the X-ray powder diffraction
spectrum thereof is obtained by using Cu-K.alpha. radiation and
represented by diffraction angle 2.theta. angle, in which there are
characteristic peaks at diffraction angle 2.theta. angles of 9.95,
12.12, 12.68, 13.56, 14.73, 15.32, 16.20, 17.27, 18.74, 20.00,
21.57, 22.82, 23.22, 24.35, 24.63, 25.42, 27.13, 28.15, 30.01,
30.30, 32.92 and 34.97, wherein the error range of 2.theta. angle
of each characteristic peak is .+-.0.2.
[0035] The present invention also provides a method for preparing
the crystal form A of the compound of formula (I''), characterized
in that the method is selected from the following preparation
methods:
[0036] (1) reacting the free form of the compound of formula (I'')
with malic acid in a solvent or a mixed solvent, followed by
stirring, precipitation, filtration and drying to obtain the
desired crystal form A; wherein the solvent is an alcohol solvent,
the mixed solvent is a mixed solvent of an alcohol and water, the
alcohol solvent is preferably isopropanol, and the mixed solvent of
an alcohol and water is preferably a mixed solvent of isopropanol
and water; and
[0037] (2) adding the compound of formula (I'') to a solvent or a
mixed solvent, followed by pulping, filtration and drying to obtain
the desired crystal form A; wherein the solvent is an alcohol
solvent, the mixed solvent is a mixed solvent of an alcohol and
water, the alcohol solvent is preferably isopropanol, and the mixed
solvent of an alcohol and water is preferably a mixed solvent of
isopropanol and water.
[0038] The present invention also relates to a pharmaceutical
composition comprising a salt, the crystal forms I and II of the
compound of formula (I), the crystal forms 1, 2 and 3 of the
compound of formula (I'), or the crystal form A of the compound of
formula (I'') and a pharmaceutically acceptable carrier.
[0039] The present invention further relates to use of the salt,
the crystal forms I and II of the compound of formula (I), the
crystal forms 1, 2 and 3 of the compound of formula (I'), the
crystal form A of the compound of formula (I'') or the
pharmaceutical composition in the preparation of a medicament for
the treatment or prevention of a disease related to renal outer
medullary potassium channel (ROMK) inhibition.
[0040] The disease of the present invention is selected from the
group consisting of hypertension and heart failure.
[0041] The research results show that crystal form I of the
compound of formula (I) prepared according to the present invention
is stable under conditions of lighting, high temperature and high
humidity. Crystal form I is also stable under conditions of
grinding, pressure and heating, which meets the medical needs of
production, transportation and storage.
DESCRIPTION OF THE DRAWINGS
[0042] FIG. 1 shows the XRPD (X-ray powder diffraction) spectrum of
crystal form I of the compound of formula (I).
[0043] FIG. 2 shows the DSC spectrum of crystal form I of the
compound of formula (I).
[0044] FIG. 3 shows the XRPD spectrum of crystal form II of the
compound of formula (I).
[0045] FIG. 4 shows the DSC spectrum of crystal form II of the
compound of formula (I).
[0046] FIG. 5 shows the XRPD spectrum of crystal form 1 of the
compound of formula (I').
[0047] FIG. 6 shows the DSC-TGA spectrum of crystal form 1 of the
compound of formula (I').
[0048] FIG. 7 shows the XRPD spectrum of crystal form 2 of the
compound of formula (I').
[0049] FIG. 8 shows the DSC-TGA spectrum of crystal form 2 of the
compound of formula (I').
[0050] FIG. 9 shows the XRPD spectrum of crystal form 3 of the
compound of formula (I').
[0051] FIG. 10 shows the XRPD spectrum of crystal form A of the
compound of formula (I'').
[0052] FIG. 11 shows the DSC-TGA spectrum of crystal form A of the
compound of formula (I'').
DETAILED DESCRIPTION OF THE INVENTION
[0053] The present invention is illustrated by the following
examples in detail. The examples of the present invention are
merely intended to describe the technical solution of the present
invention, and should not be considered as limiting the scope of
the present invention.
[0054] Testing instruments used in the experiments
[0055] 1. DSC spectrum
[0056] Instrument type: Mettler Toledo DSC 1 Staree System
[0057] Purging gas: Nitrogen
[0058] Heating rate: 10.0.degree. C./min
[0059] Temperature range: 40-350.degree. C.
[0060] 2. X-ray diffraction spectrum
[0061] Instrument type: Bruker D8 Focus X-ray powder
diffractometer
[0062] Ray: monochromatic Cu-K.alpha. ray (.lamda.=1.5406)
[0063] Scanning mode: .theta./2.theta., Scanning range:
2-40.degree.
[0064] Voltage: 40 KV, Electric current: 40 mA
Example 1
Synthesis of
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide
##STR00005##
[0065] Step 1
5-Bromo-4-methoxypicolinic acid
[0066] Methyl 5-bromo-4-methoxypicolinate 1a (250 mg, 1.01 mmol)
was dissolved in 10 mL of a mixed solvent of methanol,
tetrahydrofuran and water (V:V:V=3:3:1), sodium hydroxide (100 mg,
2.5 mmol) was added and the mixture was stirred for 2 hours. The
reaction solution was concentrated under reduced pressure, and 10
mL of water was added. The reaction solution was adjusted to pH 2
by adding 2M hydrochloric acid dropwise and extracted with ethyl
acetate (20 mL.times.3). The organic phase was washed with
saturated NaCl solution (15 mL.times.2), dried over anhydrous
sodium sulfate, and filtered. The filtrate was concentrated under
reduced pressure to obtain the crude title compound
5-bromo-4-methoxypicolinic acid 1b (200 mg, a white solid), which
was used in the next step without further purification.
[0067] MS m/z (ESI): [M-1].sup.+=229.9.
Step 2
tert-Butyl
4-(5-bromo-4-methoxypicolinamido)piperidine-1-carboxylate
[0068] Crude 5-bromo-4-methoxypicolinic acid 1b (150 mg, 0.65
mmol), tert-butyl 4-aminopiperidine-1-carboxylate (130 mg, 0.65
mmol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide (190 mg, 1
mmol), 1-hydroxybenzotriazole (20 mg, 0.13 mmol) and triethylamine
(0.15 mL, 1 mmol) were dissolved in 20 mL of N,N-dimethylformamide.
The reaction mixture was warmed up to 50.degree. C. and stirred for
6 hours. The reaction solution was concentrated under reduced
pressure. The resulting residue was purified by thin layer
chromatography with n-hexane and ethyl acetate as the developing
solvents to obtain the title compound tert-butyl
4-(5-bromo-4-methoxypicolinamido)piperidine-1-carboxylate 1c (60
mg, a light yellow oil), yield: 22.4%.
[0069] MS m/z (ESI): [M+1].sup.+=414.1.
Step 3
tert-Butyl
4-(5-cyano-4-methoxypicolinamido)piperidine-1-carboxylate
[0070] tert-Butyl
4-(5-bromo-4-methoxypicolinamido)piperidine-1-carboxylate 1c (60
mg, 0.15 mmol), zinc cyanide (26 mg, 0.22 mmol) and
tetra(phenylphosphine)palladium (18 mg, 0.015 mmol) were dissolved
in 1.5 mL of N,N-dimethylacetamide. The reaction was carried out
under microwave for 40 minutes at 135.degree. C. The reaction
solution was concentrated under reduced pressure. The resulting
residue was purified by thin layer chromatography with n-hexane and
ethyl acetate as the developing solvents to obtain the title
compound tert-butyl
4-(5-cyano-4-methoxypicolinamido)piperidine-1-carboxylate 1d (32
mg, a colorless oil), yield: 61.5%.
[0071] MS m/z (ESI): [M+1].sup.+=361.2.
Step 4
5-Cyano-4-methoxy-N-(piperidin-4-yl)picolinamide
[0072] tert-Butyl
4-(5-cyano-4-methoxypicolinamido)piperidine-1-carboxylate 1d (32
mg, 0.09 mmol) was dissolved in 5 mL of dichloromethane, and 1 mL
of trifluoroacetic acid was added. The reaction mixture was stirred
for 1.5 hours. The reaction solution was concentrated under reduced
pressure, and 15 mL of methanol was added. The reaction solution
was adjusted to pH 8 by adding saturated sodium bicarbonate
solution dropwise, and then concentrated under reduced pressure.
The resulting residue was purified by thin layer chromatography
with dichloromethane and methanol as the developing solvents to
obtain the title compound
5-cyano-4-methoxy-N-(piperidin-4-yl)picolinamide 1e (23 mg, a white
paste), yield: 100%.
[0073] MS m/z (ESI): [M+1].sup.+=261.1.
Step 5
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5-y-
l)ethyl) piperidin-4-yl)-4-methoxypicolinamide
[0074] (R)-4-methyl-5-(oxiran-2-yl)isobenzofuran-1(3H)-one (25 mg,
0.09 mmol, prepared according to the method disclosed in patent
application "WO2010129379") and
5-cyano-4-methoxy-N-(piperidin-4-yl)picolinamide 1e (23 mg, 0.09
mmol) were dissolved in 5 mL of acetonitrile. The reaction mixture
was stirred and heated to reflux for 15 hours. The reaction
solution was concentrated under reduced pressure. The resulting
residue was purified by thin layer chromatography with
dichloromethane and methanol as the developing solvents to obtain
the title compound
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide 1 (4.5 mg, a light
yellow solid), yield: 11.3%.
[0075] MS m/z (ESI): [M+1].sup.+=450.2.
[0076] .sup.1H NMR (400 MHz, d.sub.6-DMSO): .delta.=8.88 (s, 1H),
8.75 (d, 1H), 7.77 (s, 1H), 7.71-7.69 (m, 2H), 5.43-5.40 (m, 2H),
5.35 (s, 1H), 5.08 (s, 1H), 4.09 (s, 3H), 3.78 (s, 1H), 2.95 (s,
3H), 2.38 (s, 1H), 2.27 (s, 3H), 2.25 (s, 2H), 1.72 (s, 4H)
ppm.
Example 2. Preparation of Crystal Form I of the Tartrate
[0077] L-tartaric acid (0.4 g, 2.66 mmol) was added to a 50 ml
reaction flask, 30 ml of methanol was added, and then the mixture
was heated to 70.degree. C. to dissolve.
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide (1.0 g, 2.22 mmol)
(prepared according to the method of Example 1) was added, and the
reaction was carried out at 70.degree. C. for 24 hours. The mixture
was cooled to room temperature, filtered and dried to obtain 1.22 g
of a solid, yield: 91.7%. The X-ray diffraction spectrum of the
crystal sample is shown in FIG. 1. There are characteristic peaks
at about 3.49(25.27), 10.22(8.65), 12.27(7.21), 13.69(6.47),
15.46(5.73), 16.98(5.22), 18.04(4.91), 19.45(4.56), 20.89(4.25),
23.95 (3.71), 25.63(3.47), 26.71(3.34), and 29.44(3.03). The DSC
spectrum is shown in FIG. 2, having a sharp melting endothermic
peak at 218.42.degree. C. The crystal form was defined as crystal
form I.
Example 3. Preparation of Crystal Form II of the Tartrate
[0078]
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofu-
ran-5-yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide (1.0 g, 2.22
mmol) (prepared according to the method of Example 1) and
L-tartaric acid (0.4 g, 2.66 mmol) were added to a 50 ml reaction
flask, 30 ml of isopropanol/tetrahydrofuran/water (V:V:V=20:10:1)
was added, and the mixture was heated to 70.degree. C. for 24
hours. The mixture was cooled to room temperature, filtered and
dried to obtain 1.15 g of a solid, yield: 86.5%. The X-ray
diffraction spectrum of the crystal sample is shown in FIG. 3.
There are characteristic peaks at about 3.88(22.78), 13.00(6.80),
17.25(5.14), 25.42(3.50), and 27.79(3.21). The DSC spectrum is
shown in FIG. 4, having a wide melting endothermic peak at
191.09.degree. C. The crystal form was defined as crystal form
II.
Example 4. Preparation of Crystal Form I of the Tartrate
[0079] L-tartaric acid (0.4 g, 2.66 mmol) was added to a 50 ml
reaction flask, 30 ml of ethanol/water (V:V=9:1) was added, and
then the mixture was heated to 70.degree. C. to dissolve.
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide (1.0 g, 2.22 mmol)
(prepared according to the method of Example 1) was added, and then
the reaction was carried out at 70.degree. C. for 24 hours. The
mixture was cooled to room temperature, filtered and dried to
obtain 1.24 g of a solid, yield: 93.2%. The product was identified
as crystal form I after studying and comparing the X-ray
diffraction and DSC spectra.
Example 5. Preparation of Crystal Form I of the Tartrate
[0080] L-tartaric acid (0.4 g, 2.66 mmol) was added to a 50 ml
reaction flask, 30 ml of ethanol/water (V:V=19:1) was added, and
then the mixture was heated to 70.degree. C. to dissolve.
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide (1.0 g, 2.22 mmol)
(prepared according to the method of Example 1) was added, and then
the reaction was carried out at 70.degree. C. for 24 hours. The
mixture was cooled to room temperature, filtered and dried to
obtain 1.20 g of a solid, yield: 90.2%. The product was identified
as crystal form I after studying and comparing the X-ray
diffraction and DSC spectra.
Example 6. Preparation of the Maleate
[0081]
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofu-
ran-5-yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide (600 mg, 1.32
mmol) was added to 20 mL of ethanol. The mixture did not become
clear after heating to 70.degree. C. and stirring for 1 hour. The
solution became clear rapidly after addition of maleic acid (186
mg, 1.6 mmol). The mixture was stirred at 70.degree. C. for 18
hours, and a small amount of solid was precipitated. The mixture
was cooled naturally to room temperature, and a large amount of
solid was precipitated after 2 hours. The reaction solution was
filtered. The filter cake was rinsed with ethanol (2 mL.times.2),
collected and dried in a vacuum to obtain the title compound (600
mg, a white solid), yield: 79%.
[0082] The .sup.1H-NMR nuclear magnetic data showed that the molar
ratio of the main component to maleic acid in the salt was 1:1;
[0083] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 2.00 (d, 4H)
2.33 (br. s., 3H) 3.17 (br. s., 3H) 3.44 (br. s., 1H) 3.59 (br. s.,
1H) 3.74 (br. s., 1H) 4.11 (s, 4H) 4.37 (br. s., 1H) 5.28-5.52 (m,
3H) 6.02 (s, 2H) 6.35 (br. s., 1H) 7.63-7.89 (m, 3H) 8.93 (s, 1H)
9.11 (br. s., 1H) 9.58 (br. s., 1H).
Example 7. Preparation of the Hydrochloride
[0084]
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofu-
ran-5-yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide (100 mg, 0.22
mmol) was dissolved in 3 mL of ethanol, but it was not fully
dissolved after heating to 70.degree. C. for 30 minutes. 12 M
concentrated hydrochloric acid (0.02 mL, 0.29 mmol) was added
dropwise to the above solution, and there was no significant
change. The solution was heated to 70.degree. C. and stirred for 17
hours, and then a white powder appeared in the solution. The
reaction solution was filtered while it was still hot. The filter
cake was washed with ethanol (2 mL.times.2), collected and dried in
a vacuum to obtain the title compound (85 mg, a white solid),
yield: 79%.
[0085] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.91-2.19 (m,
4H) 2.32-2.40 (m, 3H) 3.07-3.29 (m, 3H) 3.41-3.56 (m, 1H) 3.76 (br.
s., 1H) 3.98-4.20 (m, 4H) 5.37-5.49 (m, 2H) 5.55 (d, 1H) 6.43 (d,
1H) 7.71-7.82 (m, 3H) 8.86-8.93 (m, 1H) 9.08 (d, 1H) 10.27 (br. s.,
1H).
Example 8. Preparation of the Sulfate
[0086]
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofu-
ran-5-yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide (150 mg, 0.33
mmol) was dissolved in a mixed solution of 8 mL of dichloromethane
and 4 mL of isopropanol, but it was not fully dissolved.
Concentrated sulfuric acid (42 mg, 0.43 mmol) was added dropwise to
the above solution, which was stirred at room temperature
(16.degree. C.) for 4 hours, and then 4 mL of methyl tert-butyl
ether were added. The reaction solution was cooled in an ice bath
to precipitate a solid, and then filtered. The filter cake was
collected and dried in a vacuum to obtain the title compound (153
mg, a white solid), yield: 85%.
[0087] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.89-2.18 (m,
4H), 2.30-2.40 (m, 3H), 3.07-3.30 (m, 3H), 3.45 (br. s., 1H), 3.66
(d, 1H), 3.75-3.85 (m, 1H), 3.97-4.19 (m, 3H), 5.31-5.53 (m, 3H),
6.28-6.48 (m, 1H), 7.71-7.83 (m, 3H), 8.83-8.96 (m, 1H), 9.10 (d,
1H), 9.51 (br. s., 1H).
Example 9. Preparation of the Citrate
[0088]
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofu-
ran-5-yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide (161 mg, 0.36
mmol) was dissolved in 5 mL of isopropanol, but it was not fully
dissolved. Citric acid (89 mg, 0.46 mmol) was added to the above
solution. The materials were not fully dissolved after heating to
reflux and stirring for 16 hours, and there were massive and
powdery solids. The mixture was filtered, and the filter cake was
collected and dried in a vacuum. The solid was crushed and mixed
well to obtain the title compound (150 mg, a white solid), yield:
65%.
[0089] The .sup.1H-NMR nuclear magnetic data showed that the molar
ratio of the main component to citric acid in the salt was 1:1.
[0090] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.83 (br. s.,
4H), 2.30 (s, 3H), 2.52-2.65 (m, 3H), 2.67 (br. s., 1H), 2.70-2.82
(m, 2H), 3.21 (br. s., 2H), 3.32 (br. s., 1H), 3.69-3.82 (m, 1H),
3.92 (br. s., 1H), 4.10 (s, 3H), 4.34 (br. s., 1H), 5.19 (d, 1H),
5.35-5.47 (m, 2H), 7.68-7.74 (m, 2H), 7.78 (s, 1H), 8.85 (d, 1H),
8.90 (s, 1H).
Example 10. Preparation of the Phosphate
[0091]
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofu-
ran-5-yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide (100 mg, 0.22
mmol) was dissolved in 3 mL of isopropanol, but it was not fully
dissolved after heating to 70.degree. C. for 30 minutes. Phosphoric
acid (33 mg, 0.29 mmol) was added dropwise to the above solution,
and then a solid was condensed and precipitated in the solution.
The mixture was heated to 70.degree. C. and stirred for 16 hours,
and the solid in the solution was powdery. The mixture was filtered
without cooling, and the filter cake was collected and washed with
isopropanol (2 mL.times.2) to obtain the title compound (110 mg, a
white solid), yield: 91%.
[0092] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.82 (br. s.,
4H), 2.29 (s, 3H), 2.50-2.78 (m, 4H), 3.18 (br. s., 2H), 3.88 (br.
s., 1H), 4.03-4.16 (m, 3H), 5.19 (d, 1H), 5.34-5.46 (m, 2H),
7.68-7.74 (m, 2H), 7.78 (s, 1H), 8.82 (d, 1H), 8.90 (s, 1H).
Example 11. Preparation of the Fumarate
[0093] Method 1.
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide (152 mg, 0.38 mmol)
was dissolved in a mixed solution of 6 mL of dichloromethane and 3
mL of methanol. Fumaric acid (51 mg, 0.44 mmol) was dissolved in
1.5 mL of ethanol, and then added dropwise to the above solution.
The solution was heated to 40.degree. C. and stirred for 18 hours,
then cooled in an ice bath, and no solid was precipitated. 6 mL of
methyl tert-butyl ether was added to the reaction solution, and
still no solid was precipitated. Another 6 mL of methyl tert-butyl
ether was added, leading to a solid precipitate. The mixture was
filtered under reduced pressure to obtain the title compound (165
mg, a white solid), yield: 77%;
[0094] The .sup.1H-NMR nuclear magnetic data showed that the molar
ratio of the main component to fumaric acid in the salt was
1:0.5;
[0095] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.64-1.80 (m,
4H), 2.20-2.36 (m, 6H), 2.41-2.48 (m, 1H), 2.52-2.60 (m, 1H), 3.00
(d, 2H), 3.17 (s, 1H), 3.80 (br. s., 1H), 4.10 (s, 3H), 5.10 (dd,
1H), 5.34-5.45 (m, 2H), 6.59 (s, 1H), 7.65-7.72 (m, 2H), 7.78 (s,
1H), 8.76 (d, 1H), 8.89 (s, 1H).
[0096] Method 2.
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofuran-5--
yl)ethyl) piperidin-4-yl)-4-methoxypicolinamide (100 mg, 0.22 mmol)
and fumaric acid (31 mg, 0.26 mmol) were dissolved in a mixed
solution of 1 mL of dioxane and 0.5 mL of acetonitrile. The
reaction solution became clear after heating to 70.degree. C. and
stirring for 18 hours. A solid was precipitated after the reaction
solution was cooled to room temperature (20.degree. C.). The
mixture was filtered under reduced pressure to obtain the title
compound (70 mg, a white solid), yield: 56%.
[0097] The .sup.1H-NMR nuclear magnetic data showed that the molar
ratio of the main component to fumaric acid in the salt was
1:1.
[0098] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.76 (br. s.,
4H), 2.25-2.38 (m, 6H), 2.54-2.62 (m, 1H), 3.04 (d, 2H) 3.81 (br.
s., 1H), 4.10 (s, 3H), 5.12 (dd, 1H), 5.34-5.47 (m, 2H), 6.60 (s,
2H), 7.64-7.74 (m, 2H), 7.78 (s, 1H), 8.77 (d, 1H), 8.89 (s,
1H).
Example 12. Preparation of the Succinate
[0099]
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofu-
ran-5-yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide (50 mg, 0.11
mmol) and succinic acid (15 mg, 0.13 mmol) were dissolved in 1.5 mL
of methanol. The materials were not fully dissolved after heating
to 60.degree. C. for 4 hours. The mixture was cooled naturally to
room temperature (19.degree. C.) and stirred for 18 hours, and then
filtered. The filter cake was washed with 1 mL of methanol, and
dried to obtain the title compound (50 mg, a white solid), yield:
81%.
Example 13. Preparation of Crystal Form 1 of the Malate
[0100]
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofu-
ran-5-yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide (50 mg, 0.11
mmol) was added to a mixed solution of 1.5 mL of methanol and 0.15
mL of water, and then malic acid (17 mg, 0.13 mmol) was added. The
mixture did not become clear after heating to 50.degree. C. and
stirring for 4 hours. The mixture was cooled naturally to room
temperature and stirred for 16 hours. The reaction solution was
filtered, and the filter cake was rinsed with ethanol (5
mL.times.2), collected and dried in a vacuum to obtain the title
compound (40 mg, a white solid), yield: 67%.
[0101] The sample has characteristic peaks at 6.85(12.9),
9.76(9.06), 11.81(7.49), 12.48(7.09), 13.52(6.54), 14.23(6.22),
15.29(5.79), 15.94(5.56), 17.55(5.05), 17.89(4.96), 19.65(4.51),
20.25(4.38), 21.50(4.13), 22.68(3.92), 23.50(3.78), 24.27(3.66),
25.22(3.53), 26.18(3.40), 27.83(3.20), 28.59(3.12), 34.88(2.57),
and 35.84(2.50). The X-ray powder diffraction spectrum (XRPD
spectrum) thereof is shown in FIG. 5. The DSC-TGA spectrum is shown
in FIG. 6, having a sharp melting endothermic peak at
137.65.degree. C. The crystal form was defined as crystal form
1.
Example 14. Preparation of Crystal Form 2 of the Malate
[0102]
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofu-
ran-5-yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide (50 mg, 0.11
mmol) was added to a mixed solution of 1.5 mL of acetonitrile and
0.15 mL of deionized water, and then malic acid (17 mg, 0.13 mmol)
was added. The mixture became clear after heating to 50.degree. C.
and stirring for 4 hours. The mixture was cooled naturally to room
temperature and stirred for 16 hours to precipitate a solid. The
reaction solution was filtered, and the filter cake was collected
and dried in a vacuum to obtain the title compound (40 mg, a white
solid), yield: 67%.
[0103] The .sup.1H-NMR nuclear magnetic data showed that the molar
ratio of the main component to malic acid in the salt was 1:1.
[0104] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.67-1.91 (m,
4H), 2.29 (s, 3H), 2.39-2.44 (m, 2H), 2.57 (dd, 2H), 2.62-2.77 (m,
2H), 3.10-3.23 (m, 2H), 3.83-3.95 (m, 1H), 4.04-4.17 (m, 4H), 5.18
(dd, 1H), 5.33-5.49 (m, 2H), 7.67-7.74 (m, 2H), 7.78 (s, 1H), 8.83
(d, 1H), 8.90 (s, 1H).
[0105] The sample has characteristic peaks at 7.30(12.00),
9.34(9.46), 10.61(8.33), 12.24(7.23), 12.78(6.92), 13.83(6.40),
14.26(6.21), 15.52(5.71), 16.33(5.42), 17.83(4.97), 18.83(4.71),
19.86(4.47), 20.55(4.32), 21.72(4.09), 22.31(3.98), 22.62(3.93),
23.42(3.80), 24.78(3.59), 25.03(3.55), 25.27(3.52), 25.78(3.45),
26.02(3.42), 26.76(3.33), 27.10(3.29), 27.99(3.19), 30.01(2.98),
30.72(2.91), 31.19(2.87), 32.41(2.76), 33.23(2.69), 33.83(2.65),
34.41(2.60), 35.48(2.53), 36.68(2.45), 38.20(2.35), 38.92(2.31),
39.87(2.26), 41.25(2.19), 41.94(2.15), 42.38(2.13), 44.25(2.05),
and 45.72(1.98). The X-ray powder diffraction spectrum (XRPD
spectrum) thereof is shown in FIG. 7. The DSC-TGA spectrum is shown
in FIG. 8, having a sharp melting endothermic peak at
187.90.degree. C. The crystal form was defined as crystal form
2.
Example 15. Preparation of Crystal Form 3 of the Malate
[0106]
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofu-
ran-5-yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide (50 mg, 0.11
mmol) was added to 1.5 mL of isopropanol, and then malic acid (17
mg, 0.13 mmol) was added. The mixture did not become clear after
heating to 60.degree. C. and stirring for 4 hours. The mixture was
cooled naturally to room temperature and stirred for 16 hours to
precipitate a colloidal solid. The reaction solution was filtered,
and the filter cake was rinsed with isopropanol (1 mL.times.1),
collected and dried in a vacuum to obtain the title compound (50
mg, a white solid), yield: 80%;
[0107] The sample has characteristic peaks at 7.07(12.49),
9.34(9.47), 10.63(8.31), 12.79(9.92), 14.22(6.22), 15.50(5.71),
17.84(4.97), 18.86(4.70), 19.93(4.45), 22.56(3.94), 23.51(3.78),
25.16(3.54), and 30.90(2.89). The X-ray powder diffraction spectrum
(XRPD spectrum) thereof is shown in FIG. 9. The crystal form was
defined as crystal form 3.
Example 16. Preparation of Crystal Form A of the Malate
[0108]
(R)-5-cyano-N-(1-(2-hydroxy-2-(4-methyl-1-oxo-1,3-dihydroisobenzofu-
ran-5-yl)ethyl)piperidin-4-yl)-4-methoxypicolinamide (50 mg, 0.11
mmol) was added to a mixed solution of 1.5 mL of isopropanol and
0.15 mL of water, and then malic acid (17 mg, 0.13 mmol) was added.
The mixture did not become clear after heating to 50.degree. C. and
stirring for 4 hours. The mixture was cooled naturally to room
temperature and stirred for 16 hours. The reaction solution was
filtered, and the filter cake was rinsed with ethanol (5
mL.times.2), collected and dried in a vacuum to obtain the title
compound (40 mg, a white solid), yield: 67%.
[0109] The .sup.1H-NMR nuclear magnetic data showed that the molar
ratio of the main component to malic acid in the salt was
1:0.5.
[0110] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.73 (br. s.,
1H) 1.78 (br. s., 3H) 2.29 (s, 3H) 2.33-2.44 (m, 2H) 2.55 (dd, 2H)
2.60-2.68 (m, 1H) 3.09 (br. s., 2H) 3.86 (br. s., 1H) 4.00-4.16 (m,
4H) 5.14 (d, 1H) 5.33-5.46 (m, 2H) 7.66-7.74 (m, 2H) 7.78 (s, 1H)
8.80 (d, 1H) 8.89 (s, 1H).
[0111] The sample has characteristic peaks at 7.36(12.00),
9.95(8.88), 12.12(7.30), 12.68(6.97), 13.56(6.53), 14.73(6.01),
15.32(5.78), 16.20(5.47), 17.27(5.13), 18.74(4.73), 20.00(4.44),
21.57(4.12), 22.21(4.00), 22.82(3.89), 23.22(3.83), 24.35(3.65),
24.63(3.61), 25.42(3.50), 26.57(3.35), 27.13(3.28), 28.15(3.17),
29.45(3.03), 30.01(2.98), 30.30(2.95), 31.09(2.87), 32.92(2.72),
34.01(2.63), 34.97(2.56), 38.17(2.36), 40.33(2.24), and
43.19(2.09). The X-ray powder diffraction spectrum (XRPD spectrum)
thereof is shown in FIG. 10. The DSC-TGA spectrum is shown in FIG.
11, having a sharp melting endothermic peak at 144.79.degree. C.
The crystal form was defined as crystal form A.
Example 17
[0112] The sample of crystal form I of the compound of formula (I)
prepared in Example 2 and the sample of crystal form II prepared in
Example 3 were spread flat in the air respectively to test the
stability under conditions of lighting (4500 Lux), heating
(40.degree. C., 60.degree. C.), and high humidity (RH 75%, RH 90%).
Samplings were carried out on Day 5 and Day 10. The purity as
detected by HPLC is shown in Table 1.
TABLE-US-00001 TABLE 1 Stability comparison of the samples of
crystal forms I and II of the compound of formula (I) Sample Time
name (day) Lighting 40.degree. C. 60.degree. C. RH 75% RH 90%
Crystal 0 99.67% 99.67% 99.67% 99.67% 99.67% form I 5 99.54% 99.52%
99.55% 99.53% 99.59% 10 99.34% 99.45% 99.39% 99.39% 99.43% Crystal
0 99.47% 99.47% 99.47% 99.47% 99.47% form II 5 99.23% 99.38% 99.22%
99.30% 99.30% 10 98.62% 99.15% 98.87% 99.10% 99.10%
[0113] The results of the stability study showed that the stability
of the sample of crystal form I of the compound of formula (I) was
significantly better than that of the sample of crystal form II
when they were spread flat in the air respectively under conditions
of lighting, high temperature and high humidity.
Example 18
[0114] Crystal form I of the compound of formula (I) prepared
according to the method of Example 2 was ground, heated and
tableted. The results showed that the crystal form was stable. The
detailed experimental data are shown in Table 2 below.
TABLE-US-00002 TABLE 2 Special stability study of crystal form I of
the compound of formula (I) Treatment Crystal Sample Process
Experimental procedure form DSC peak Crystal Grinding 1 g of the
sample of crystal form I Crystal 214.17.degree. C. form I treatment
for of the compound of formula (I) was form I 10 minutes ground for
10 minutes in a mortar under nitrogen atmosphere. Crystal Heating 1
g of the sample of crystal form I Crystal 214.40.degree. C. form I
treatment for of the compound of formula (I) was form I 3 hours at
spread flat and heated at 80.degree. C. for 3 80.degree. C. hours.
Crystal Tableting The sample of crystal form I of the Crystal
214.44.degree. C. form I treatment compound of formula (I) was form
I tableted.
Example 19. Pharmacokinetics Assay of Crystal Form 2 of the Malate
of the Present Invention and Free Form in Rats
[0115] SD rats were used as test animals. The drug concentration in
plasma at different time points was determined by LC/MS/MS method
after intragastrical administration of the crystal form 2 of the
malate of the present invention and free form. The pharmacokinetic
behavior and property of the crystal form 2 of the malate of the
present invention and free form were studied and evaluated in SD
rats.
[0116] Samples: crystal form 2 of the malate of the present
invention and free form (the product of Example 1), prepared
according to the methods of Examples 14 and 1.
[0117] Test animals: 8 healthy SD rats, half male and half female,
were divided in two groups, and purchased from SINO-BRITSH SIPPR/BK
LAB. ANIMAL LTD., CO, with certificate No.: SOCK (Shanghai)
2008-0016.
[0118] Preparation of the test compounds: the test compounds were
added to 0.5% CMC-Na to prepare 0.5 mg/mL homogeneous suspensions
for oral administration by ultrasound.
[0119] Administration: after an overnight fasting, 8 healthy SD
rats, half male and half female, were administered the test
compounds intragastrically at an administration volume of 10
mL/kg.
[0120] Method:
[0121] After an overnight fasting, 8 healthy SD rats, half male and
half female, were administered the test compounds intragastrically.
Blood samples (0.1 mL) were taken by jugular vein puncture before
administration and at 0.5, 1, 2, 4, 6, 8, 12, and 24 hours after
administration, and heparin sodium was used as an anticoagulant.
The blood samples were centrifuged for 10 minutes at 3,500 rpm to
separate the blood plasma, which was stored at -20.degree. C. The
content of the test compound in the plasma of SD rats after
intragastric administration was determined by LC/MS/MS method.
[0122] Experimental Results
TABLE-US-00003 TABLE 3 Results of pharmacokinetics assay in SD rats
(po: 5.0 mg/kg) AUC.sub.last Cl/F.sub.--.sub.obs
Vz/F.sub.--.sub.obs Test sample T.sub.1/2(h) (ng/mL h*) (mL/min/kg)
(mL/kg) Crystal form 2 5.24 13365 6.87 3089 of the malate Free form
3.62 9283 9.82 3019
Among them, T.sub.1/2 refers to half-life, AUC.sub.last refers to
area under curve (0.fwdarw.t), Cl/F refers to clearance, and Vz/F
refers to apparent distribution volume.
Experimental Conclusion
[0123] It can be seen from the results in Table 3 that compared to
the free form, the crystal form 2 of the malate of the present
invention has longer half-life, lower clearance, and higher
exposure dose, indicating that the crystal form 2 of the malate of
the present invention has good pharmacokinetic properties.
* * * * *