U.S. patent application number 14/306261 was filed with the patent office on 2014-10-02 for novel urea compounds.
The applicant listed for this patent is Eli Lilly and Company. Invention is credited to Fucheng QU.
Application Number | 20140296171 14/306261 |
Document ID | / |
Family ID | 49485785 |
Filed Date | 2014-10-02 |
United States Patent
Application |
20140296171 |
Kind Code |
A1 |
QU; Fucheng |
October 2, 2014 |
NOVEL UREA COMPOUNDS
Abstract
The present invention provides a compound of Formula I:
##STR00001## or a pharmaceutically acceptable salt thereof.
Inventors: |
QU; Fucheng; (Carmel,
IN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Eli Lilly and Company |
Indianapolis |
IN |
US |
|
|
Family ID: |
49485785 |
Appl. No.: |
14/306261 |
Filed: |
June 17, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14035997 |
Sep 25, 2013 |
8785404 |
|
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14306261 |
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61710123 |
Oct 5, 2012 |
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Current U.S.
Class: |
514/27 |
Current CPC
Class: |
A61K 31/70 20130101;
C07H 17/02 20130101; C07D 487/10 20130101; A61K 31/155 20130101;
A61P 3/10 20180101; A61K 31/706 20130101 |
Class at
Publication: |
514/27 |
International
Class: |
A61K 31/706 20060101
A61K031/706; A61K 31/155 20060101 A61K031/155 |
Claims
1-4. (canceled)
5. A method of treating diabetes in a patient comprising
administering to a patient in need of such treatment an effective
amount of a compound of the formula: ##STR00022## or a
pharmaceutically acceptable salt thereof, in combination with an
effective amount of metformin
6. A method of treating diabetes in a patient comprising
administering to a patient in need of such treatment an effective
amount of a compound of the formula: ##STR00023## in combination
with an effective amount of metformin.
Description
[0001] The present invention relates to novel urea compounds, to
pharmaceutical compositions comprising the compounds, to methods of
using the compounds to treat physiological disorders, and to
intermediates and processes useful in the synthesis of the
compounds.
[0002] The present invention is in the field of treatment of
diabetes and other diseases and disorders associated with
hyperglycemia. Diabetes is a group of diseases that is
characterized by high levels of blood glucose. It affects
approximately 25 million people in the United States and is also
the 7.sup.th leading cause of death in U.S. according to the 2011
National Diabetes Fact Sheet (U.S. Department of Health and Human
Services, Centers for Disease Control and Prevention).
Sodium-coupled glucose cotransporters (SGLT's) are one of the
transporters known to be responsible for the absorption of
carbohydrates, such as glucose. More specifically, SGLT1 is
responsible for the transport of glucose across the brush border
membrane of the small intestine Inhibition of SGLT1 may result in
reduced absorption of glucose in the small intestine, thus
providing a useful approach to treating diabetes.
[0003] U.S. Pat. No. 7,655,632 discloses certain pyrazole
derivatives with human SGLT1 inhibitory activity which are further
disclosed as useful for the prevention or treatment of a disease
associated with hyperglycemia, such as diabetes. In addition, WO
2011/039338 discloses certain pyrazole derivatives with SGLT1/SGLT2
inhibitor activity which are further disclosed as being useful for
treatment of bone diseases, such as osteoporosis.
[0004] There is a need for alternative drugs and treatment for
diabetes. The present invention provides novel inhibitors of SGLT1
which may be suitable for the treatment of diabetes.
[0005] Accordingly, the present invention provides a compound of
Formula I:
##STR00002##
or a pharmaceutically acceptable salt thereof.
[0006] The present invention also provides a method of treating
diabetes in a patient comprising administering to a patient in need
of such treatment an effective amount of a compound of Formula I,
or a pharmaceutically acceptable salt thereof. The present
invention further provides a method of treating type 1 diabetes in
a patient comprising administering to a patient in need of such
treatment an effective amount of a compound of Formula I, or a
pharmaceutically acceptable salt thereof. In addition, the present
invention provides a method of treating type 2 diabetes in a
patient comprising administering to a patient in need of such
treatment an effective amount of a compound of Formula I, or a
pharmaceutically acceptable salt thereof. The present invention
also provides a method of treating impaired glucose tolerance
(IGT), impaired fasting glucose (IFG), or metabolic syndrome in a
patient comprising administering to a patient in need of such
treatment an effective amount of a compound of Formula I, or a
pharmaceutically acceptable salt thereof.
[0007] Furthermore, this invention provides a compound of Formula I
or a pharmaceutically acceptable salt thereof for use in therapy,
in particular for the treatment of diabetes. In addition, this
invention provides a compound of Formula I or a pharmaceutically
acceptable salt thereof for use in the treatment of type 1
diabetes. In addition, this invention provides a compound of
Formula I or a pharmaceutically acceptable salt thereof for use in
the treatment of type 2 diabetes. This invention also provides the
use of a compound of Formula I, or a pharmaceutically acceptable
salt thereof, for the manufacture of a medicament for the treatment
of diabetes. Furthermore, this invention provides the use of a
compound of Formula I, or a pharmaceutically acceptable salt
thereof, for the manufacture of a medicament for the treatment of
type 1 diabetes. This invention also provides the use of a compound
of Formula I, or a pharmaceutically acceptable salt thereof, for
the manufacture of a medicament for the treatment of type 2
diabetes. The invention also provides the use of a compound of
Formula I, or a pharmaceutically acceptable salt thereof, for the
manufacture of a medicament for the treatment of IGT, IFG, or
metabolic syndrome.
[0008] The invention further provides a pharmaceutical composition
comprising a compound of Formula I, or a pharmaceutically
acceptable salt thereof, in combination with one or more
pharmaceutically acceptable carriers, diluents, or excipients. This
invention also encompasses novel intermediates and processes for
the synthesis of the compound of Formula I.
[0009] As used herein, the terms "treating" or "to treat" includes
prohibiting, restraining, slowing, stopping, or reversing the
progression or severity of an existing symptom or disorder.
[0010] As used herein, the term "patient" refers to a mammal, such
as a mouse, guinea pig, rat, dog, or human. It is understood that
the preferred patient is a human.
[0011] As used herein, the term "effective amount" refers to the
amount or dose of compound of the invention, or a pharmaceutically
acceptable salt thereof which, upon single or multiple dose
administration to the patient, provides the desired effect in the
patient under diagnosis or treatment.
[0012] An effective amount can be readily determined by the
attending diagnostician, as one skilled in the art, by the use of
known techniques and by observing results obtained under analogous
circumstances. In determining the effective amount for a patient, a
number of factors are considered by the attending diagnostician,
including, but not limited to: the species of mammal; its size,
age, and general health; the specific disease or disorder involved;
the degree of or involvement or the severity of the disease or
disorder; the response of the individual patient; the particular
compound administered; the mode of administration; the
bioavailability characteristics of the preparation administered;
the dose regimen selected; the use of concomitant medication; and
other relevant circumstances.
[0013] The compounds of Formula I are generally effective over a
wide dosage range. For example, dosages per day normally fall
within the range of about 0.01 to about 30 mg/kg of body weight. In
some instances dosage levels below the lower limit of the aforesaid
range may be more than adequate, while in other cases still larger
doses may be employed without causing any harmful side effect, and
therefore the above dosage range is not intended to limit the scope
of the invention in any way.
[0014] The compounds of the invention are preferably formulated as
pharmaceutical compositions administered by any route which makes
the compound bioavailable. Most preferably, such compositions are
for oral administration. Such pharmaceutical compositions and
processes for preparing same are well known in the art. (See, e.g.,
Remington: The Science and Practice of Pharmacy (D. B. Troy,
Editor, 21st Edition., Lippincott, Williams & Wilkins,
2006).
[0015] In a further aspect of the invention, the present compounds
are administered in combination with one or more therapeutic
agents, such as antidiabetic agents. Administration in combination
includes simultaneous or sequential administration. In addition,
simultaneous administration of the combination can be as a single
combination dose or separate doses of each therapeutic agent.
Examples of antidiabetic agents include metformin; a DPPIV
inhibitor, such as sitagliptin or linagliptin; a sulfonylurea, such
as glimepiride; a thiazolidinedione, such as pioglitazone; a basal
insulin, such as glargine; a rapid acting insulin, such as HUMALOG
or NOVOLOG; A GLP-1 agonist, such as exenatide or liraglutide; an
SGLT2 inhibitor, such as dapagliflozin or empagliflozin; a glucagon
receptor antagonist, such as LY2409021; and the like.
[0016] Compounds of Formula I are prepared as illustrated in the
preparations, examples, and schemes below. The reagents and
starting materials are readily available to one of ordinary skill
in the art. All substituents, unless otherwise specified are as
previously defined. It is understood that these schemes,
preparations, and examples are not intended to be limiting to the
scope of the invention in any way.
[0017] Examples of resolutions include selective crystallization
techniques or chiral chromatography. (See, e.g. J. Jacques, et al.,
"Enantiomers, Racemates, and Resolutions", John Wiley and Sons,
Inc., 1981, and E. L. Eliel and S. H. Wilen," Stereochemistry of
Organic Compounds", Wiley-Interscience, 1994). It should be further
clear to one of ordinary skill in the art that separation and
isolation, by chromatography, chiral chromatography or selective
crystallization, of individual diastereomers or geometric isomers
of Formula I or individual diastereomers or geometric isomers of
intermediates leading to Formula I, can occur at any convenient
point in the synthesis.
[0018] As used herein, ".delta."refers to part per million
down-field from tetramethylsilane; "min" refers to minute or
minutes; "THF" refers to tetrahydrofuran; "MeOH" refers to methanol
or methyl alcohol; "HPLC" refers to high-performance liquid
chromatography; The term "Ac" refers to an acetyl substituent of
the following structure:
##STR00003##
The term "BOC" refers to a t-butyloxycarbonyl protecting group.
[0019] Pharmaceutically acceptable salts and common methodology for
preparing them are well known in the art. See, e.g., Gould, P. L.,
"Salt selection for basic drugs," International Journal of
Pharmaceutics, 33: 201-217 (1986); Bastin et al. "Salt Selection
and Optimization Procedures for Pharmaceutical New Chemical
Entities," Organic Process Research and Development, 4: 427-435
(2000); and S. M. Berge, et al., "Pharmaceutical Salts," Journal of
Pharmaceutical Sciences, Vol. 66, No. 1, January 1977. One skilled
in the art of synthesis will appreciate that the compounds of
Formula I as amines are organic bases, and that they are readily
converted to and isolated as pharmaceutically acceptable salts,
such as tartrate or HCl salts, using techniques and conditions well
known to one of ordinary skill in the art.
##STR00004## ##STR00005##
Preparation 1
(4-bromo-2-methyl-phenyl)methanol
##STR00006##
[0021] Scheme 1, step A: Add borane-tetrahydrofuran complex (0.2
mol, 200 mL, 1.0 M solution) to a solution of
4-bromo-2-methylbenzoic acid (39 g, 0.18 mol) in tetrahydrofuran
(200 mL). After 18 hours at room temperature, remove the solvent
under reduced pressure to provide a solid. Purify by flash
chromatography to yield the title compound as a white solid (32.9
g, 0.16 mol). .sup.1H NMR (CDCl3): .delta. 1.55 (s, 1H), 2.28 (s,
3H), 4.61 (s, 2H), 7.18-7.29 (m, 3H).
Alternative Synthesis of (4-bromo-2-methyl-phenyl)methanol
[0022] Scheme 1, step A: Borane-dimethyl sulfide complex (2M in
THF; 1150 mL, 2.3 mol) is added over 1.5 hours to a solution of
4-bromo-2-methylbenzoic acid (250 g, 1.16 mol) in anhydrous
tetrahydrofuran (1500 mL) at 0.degree. C. The reaction is allowed
to warm slowly to ambient temperature and stirred overnight. The
solution is cooled to -10.degree. C. and water (500 mL) is added
very slowly. Further water (5000 mL) is added and the mixture is
extracted with ethyl acetate (2.times.5000 mL). The combined
organic layers are washed with saturated aqueous NaCl solution
(5000 mL) and dried over Na.sub.2SO.sub.4. Filtration and
concentration under reduced pressure provides the title compound
(226 g, 97% yield).
Preparation 2
4-bromo-1-chloromethyl-2-methyl-benzene
##STR00007##
[0024] Scheme 1, step B: Add thionyl chloride (14.31 mL, 0.2 mol)
to a solution of (4-bromo-2-methyl-phenyl)methanol (32.9 g, 0.16
mol) in dichloromethane (200 mL) and dimethylformamide (0.025 mol,
2.0 mL) at 0.degree. C. After 1 hour at room temperature, pour the
mixture into ice-water (100 g), extract with dichloromethane (300
mL), wash extract with 5% aq. sodium bicarbonate (30 mL) and brine
(200 mL), dry over sodium sulfate, filter, and concentrate under
reduced pressure to provide the crude title compound as a white
solid (35.0 g, 0.16 mol). The material is used in the next step
without further purification. .sup.1H NMR (CDCl.sub.3): .delta.
2.38 (s, 3H), 4.52 (s, 2H), 7.13-7.35 (m, 3H).
Alternative synthesis of
4-bromo-1-(chloromethyl)-2-methyl-benzene
[0025] Scheme 1, step B: Methanesulfonyl chloride (171 mL, 2.11
mol) is added over 30 minutes to a mixture of
(4-bromo-2-methyl-phenyl)methanol (250 g, 1.24 mol) and
triethylamine (304 mL; 2.11 mol) in dichloromethane (2500 mL)
cooled in ice/water. The mixture is allowed to warm to ambient
temperature and is stirred for 16 hours. Water (5000 mL) is added
and the product is extracted with dichloromethane (2.times.7000
mL). The combined organic layers are washed with saturated aqueous
NaCl solution (5000 mL) and dried over Na.sub.2SO.sub.4. Filtration
and concentration under reduced pressure provides a residue which
is passed through a silica pad (eluting with hexane and ethyl
acetate) to provide the title compound (234 g; 86% yield).
Preparation 3
4-[(4-bromo-2-methyl-phenyl)methyl]-5-isopropyl-1H-pyrazol-3-ol
##STR00008##
[0027] Scheme 1, step C: Add sodium hydride (8.29 g, 0.21 mol, 60%
dispersion in oil) to a solution of methyl 4-methyl-3-oxovalerate
(27.1 mL, 0.19 mol) in tetrahydrofuran at 0.degree. C. After 30 min
at room temperature, add a solution of
4-bromo-1-chloromethyl-2-methyl-benzene (35.0 g, 0.16 mol) in
tetrahydrofuran (50 mL). Heat the resulting mixture at 70.degree.
C. overnight (18 hours). Add 1.0 M HCl (20 mL) to quench the
reaction. Extract with ethyl acetate (200 mL), wash the extract
with water (200 mL) and brine (200 mL), dry over Na.sub.2SO.sub.4,
filter, and concentrate under reduced pressure. Dissolve the
resulting residue in toluene (200 mL) and add hydrazine monohydrate
(23.3 mL, 0.48 mol). Heat the mixture at 120.degree. C. for 2 hours
with a Dean-Stark apparatus to remove water. Cool and remove the
solvent under the reduced pressure, dissolve the residue with
dichloromethane (50 mL) and methanol (50 mL). Pour this solution
slowly into a beaker with water (250 mL). Collect the resulting
precipitated product by vacuum filtration. Dry in vacuo in an oven
overnight at 40.degree. C. to yield the title compound as a solid
(48.0 g, 0.16 mol). MS (m/z): 311.0 (M+1), 309.0 (M-1).
Alternative Synthesis of
4-[(4-bromo-2-methyl-phenyl)methyl]-5-isopropyl-1H-pyrazol-3-ol
[0028] Scheme 1, step C: A solution of
4-bromo-1-(chloromethyl)-2-methyl-benzene (500 g, 2.27 mol) in
acetonitrile (2500 mL) is prepared. Potassium carbonate (941 g,
6.81 mol) and potassium iodide (450 g, 2.72 mol) are added and the
mixture is stirred for 2 hours. Methyl 4-methyl-3-oxovalerate (340
mL; 2.39 mmol) is added. The resulting mixture is stirred at
ambient temperature for 16 hours. Hydrochloric acid (2N; 8000 mL)
is added to give pH 3. The solution is extracted with ethyl acetate
(2.times.7000 mL), the organic phase is washed with brine (5000
mL), and dried over Na.sub.2SO.sub.4. The mixture is filtered and
concentrated under reduced pressure. The residue is dissolved in
toluene (2500 mL) and hydrazine monohydrate (340 mL, 6.81 mol) is
added. The resulting mixture is heated at 110.degree. C. and water
is removed using a Dean-Stark apparatus. After 2 hours the mixture
is cooled to 90.degree. C. and additional hydrazine monohydrate
(340 mL, 6.81 mol) is added, and the mixture is heated at
110.degree. C. for 2 hours. The mixture is cooled and concentrated
under reduced pressure. The residue is stirred with water (2500 mL)
for 1 hour and the resulting solid is filtered, then triturated in
hexane, and filtered to provide the title compound (460 g; 65%
yield). Mass spectrum (m/z): 309/311 (M+1).
Preparation 4
4-(4-bromo-2-methylbenzyl)-5-(propan-2-yl)-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-beta-D-glucopyranoside
##STR00009##
[0030] Scheme 1, step D: To a 1 L flask, add
4-[(4-bromo-2-methyl-phenyl)methyl]-5-isopropyl-1H-pyrazol-3-ol (24
g, 77.6 mmol), 2,3,4,6-tetra-O-acetyl-alpha-D-glucopyranosyl
bromide (50.4 g, 116 mmol), benzyltributylammonium chloride (5 g,
15.5 mmol), dichloromethane (250 mL), potassium carbonate (32 g,
323 mmol), and water (120 mL). Stir the reaction mixture overnight
at room temperature. Extract with dichloromethane (450 mL). Wash
the extract with water (300 mL) and brine (500 mL). Dry the organic
phase over sodium sulfate, filter, and concentrate under reduced
pressure. Purify the resulting residue by flash chromatography
(silica gel, gradient ethyl acetate/dichloromethane from 10-70%
over 20 min, 330 g column) to provide the title compound (36.5 g,
57 mmol). MS (m/z): 638.5 (M+1), 636.5 (M-1).
Alternative Synthesis of
4-(4-bromo-2-methylbenzyl)-5-(propan-2-yl)-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-beta-D-glucopyranoside
[0031] Scheme 1, step D:
4-[(4-Bromo-2-methyl-phenyl)methyl]-5-isopropyl-1H-pyrazol-3-ol
(110 g, 356 mmol), 2,3,4,6-tetra-O-acetyl-alpha-D-glucopyranosyl
bromide (204.8 g, 498 mmol), benzyltributylammonium chloride (17.2
g, 53.4 mmol), potassium carbonate (122.9 g, 889 mmol),
dichloromethane (1100 mL), and water (330 mL) are combined and the
mixture is stirred at 50.degree. C. temperature for 16 hours.
Further 2,3,4,6-tetra-O-acetyl-alpha-D-glucopyranosyl bromide
(29.26 g, 71.2 mmol) is added and the mixture stirred at 55.degree.
C. for 3 hours. The mixture is cooled to ambient temperature and
stirred overnight. Further
2,3,4,6-tetra-O-acetyl-alpha-D-glucopyranosyl bromide (29.26 g,
71.15 mmoles) is added and the mixture is heated to reflux for 1.5
hours. The mixture is cooled to ambient temperature and water (2000
mL) is added. The phases are separated and the aqueous is extracted
with dichloromethane (500 mL). The combined organic layers are
washed with water (2000 mL) and saturated aqueous NaCl solution
(2000 mL). The solution is dried over MgSO.sub.4 and filtered. The
filtrate is concentrated under reduced to provide a solution of
approximately 500 mL. This solution is poured onto a silica column
and is purified by flash chromatography (5 Kg silica), eluting
first with 100% dichloromethane, then with 40% ethyl
acetate/dichloromethane, to provide the title compound (175 g, 77%
yield). Mass spectrum (m/z): 639/641 (M+1).
Preparation 5
4-{4-[(1E)-4-hydroxybut-1-en-1-yl]-2-methylbenzyl}-5-(propan-2-yl)-1H-pyra-
zol-3-yl 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranoside
##STR00010##
[0033] Scheme 1, step E: Add 3-buten-1-ol (6.1 mL, 70 mmol) to a
solution of
4-(4-bromo-2-methylbenzyl)-5-(propan-2-yl)-1H-pyrazol-3-yl2,3,4,6-tetr-
a-O-acetyl-beta-D-glucopyranoside (15 g, 23.5 mmol) in acetonitrile
(200 mL) and triethylamine (50 mL). Degas the solution with
nitrogen over 10 minutes. Add tri-o-tolylphosphine (1.43 g, 4.7
mmol) and palladium acetate (526 mg, 2.35 mmol). After refluxing at
90.degree. C. for 2 hours, cool and concentrate to remove the
solvent under reduced pressure. Purify the resulting residue by
flash chromatography (silica gel, gradient ethyl acetate/hexanes
from 20-80% over 20 min, 330 g column) to provide the title
compound (7.5 g, 11.9 mmol). MS (m/z): 631.2 (M+1), 629.2
(M-1).
Preparation 6
(3E)-4-[3-methyl-4-({5-(propan-2-yl)-3-[(2,3,4,6-tetra-O-acetyl-beta-D-glu-
copyranosyl)oxy]-1H-pyrazol-4-yl}methyl)phenyl]but-3-en-1-yl
methanesulfonate
##STR00011##
[0035] Scheme 1, step F: Add methanesulfonyl chloride (1.35 g, 11.8
mmol) to a solution of
4-{4-[(1E)-4-hydroxybut-1-en-1-yl]-2-methylbenzyl}-5-(propan-2-yl)-1H-pyr-
azol-3-yl 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranoside (1.5 g,
2.38 mmol) and triethylamine (5 g, 49.2 mmol) in dichloromethane
(50 mL) at 0.degree. C. After 30 minutes at room temperature,
extract with dichloromethane (80 mL), and wash with water (80 mL)
and brine (40 mL). Dry organic phase over sodium sulfate, filter
and concentrate under reduced pressure. Purify the resulting
residue by flash chromatography (silica gel, gradient ethyl
acetate/dichloromethane from 10-65% over 20 min, 120 g column) to
yield the title compound (5.4 g, 7.62 mmol). MS (m/z): 708.5(M+1),
706.5 (M-1).
##STR00012##
Preparation 7
tert-butyl
4-but-3-ynyl-4,9-diazaspiro[5.5]undecane-9-carboxylate
##STR00013##
[0037] Scheme 2, step A: A mixture of tert-butyl
4,9-diazaspiro[5.5]undecane-9-carboxylate hydrochloride (170.00 g,
584.53 mmoles), cesium carbonate (476.13 g, 1.46 moles), and
4-bromobutyne (93.28 g, 701.43 mmoles) in acetonitrile (1.70 L) is
heated to 60.degree. C. and stirred at this temperature overnight.
Further 4-bromobutyne (46.64 g, 350.72 mmoles) is added and the
mixture is stirred at 60.degree. C. overnight. The mixture is
cooled and filtered. The filtrate is concentrated under reduced
pressure and the residue dissolved in ethyl acetate (500 mL),
washed with water (1 L), and saturated aqueous sodium chloride (1
L), then dried over MgSO.sub.4, and filtered. The filtrate is
concentrated under reduced pressure and the residue is purified by
silica column chromatography eluting with 20 to 70% ethyl acetate
in dichloromethane to provide the title compound (129 g, 72%
yield). Mass spectrum (m/z): 307.25 (M+1).
Preparation 8
tert-butyl 4-[(E)-4-(4,4,5,
5-tetramethyl-1,3,2-dioxaborolan-2-yl)but-3-enyl]-4,9-diazaspiro[5.5]unde-
cane-9-carboxylate
##STR00014##
[0039] Scheme 2, step B: Tert-butyl
8-but-3-ynyl-3,8-diazaspiro[5.5]undecane-3-carboxylate (129.00 g,
420.95 mmoles), triethylamine (5.87 mL, 42.10 mmoles), and
zirconocene chloride (10.86 g, 42.10 mmoles) are combined. To this
is added 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (64.13 mL 442.00
mmoles) via syringe. The reaction is then heated to 65.degree. C.
and stirred at this temperature for 24 hours. The heat is turned
off and the reaction allowed to stir at ambient temperature for a
further 24 hours. Dichloromethane (500 mL) is added and the
resulting solution is filtered through a 10 cm pad of silica
eluting with 20% ethyl acetate in dichloromethane (3.times.500 mL).
The filtrate is concentrated under reduced pressure to provide the
title compound (163.5 g; Yield 89%). Mass spectrum (m/z): 435.35
(M+1).
##STR00015## ##STR00016##
Preparation 9
tert-butyl
2-{(3E)-4-[3-methyl-4-({5-(propan-2-yl)-3-[(2,3,4,6-tetra-O-ace-
tyl-beta-D-glucopyranosyl)oxy]-1H-pyrazol-4-yl}methyl)phenyl]but-3-en-1-yl-
}-2,9-diazaspiro[5.5]undecane-9-carboxylate
##STR00017##
[0041] Scheme 3, step A: Heat a mixture of
(3E)-4-[3-methyl-4-({5-(propan-2-yl)-3-[(2,3,4,6-tetra-O-acetyl-beta-D-gl-
ucopyranosyl)oxy]-1H-pyrazol-4-yl}methyl)phenyl]but-3-en-1-yl
methanesulfonate (1.0 g, 1.41 mmol), tert-butyl
2,9-diazaspiro[5.5]undecane-9-carboxylate hydrochloride (451 mg,
1.55 mmol), and diisopropylethylamine (730 mg, 5.64 mmol) in
acetonitrile (4 mL) at 80.degree. C. overnight. Remove the solvent
under the reduced pressure. Purify the resulting residue by flash
chromatography (silica gel, gradient ethyl acetate/dichloromethane
from 25-85% over 15 min, then with methanol/dichloromethane from
1-3% over 10 min, 40 g column) to provide the title compound (630
mg, 0.73 mmol). MS (m/z): 867.2, 868.4 (M+1), 865.2, 866.4
(M-1).
Alternative Synthesis of tert-butyl
2-{(3E)-4-[3-methyl-4-({5-(propan-2-yl)-3-[(2,3,4,6-tetra-O-acetyl-beta-D-
-glucopyranosyl)oxy]-1H-pyrazol-4-yl}methyl)phenyl]but-3-en-1-yl}-2,9-diaz-
aspiro[5.5]undecane-9-carboxylate
[0042] Scheme 3, step A: A flask is charged with
4-(4-bromo-2-methylbenzyl)-5-(propan-2-yl)-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyl-beta-D-glucopyranoside (35.00 g, 54.73
mmoles), tert-butyl
8-[(E)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)but-3-enyl]-3,8-dia-
zaspiro[5.5]undecane-3-carboxylate (29.72 g, 68.41 mmoles),
potassium carbonate (22.69 g, 164.19 mmoles), tetrahydrofuran
(350.00 mL), and water (70.00 mL). The resulting solution is
degassed. Pd(OAc).sub.2 (245.75 mg, 1.09 mmoles) and
2-dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl (1.04
g, 2.19 mmoles) are then added and the solution is degassed again.
The mixture is heated to reflux and stirred overnight. The mixture
is cooled to room temperature and concentrated under reduced
pressure. Water (400 mL) is added to the residue and the mixture
extracted with ethyl acetate (2.times.500 mL then 200 mL). The
combined organic extracts are washed with brine, dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure
to provide the title compound (50.0 g .about.about 80% purity).
Mass spectrum (m/z): 867 (M+1). The compound can be purified by
flash column chromatography (silica gel) eluting with ethyl
acetate.
Preparation 10
4-{4-[(1E)-4-(2,9-diazaspiro[5.5]undec-2-yl)but-1-en-1-yl]-2-methylbenzyl}-
-5-(propan-2-yl)-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyyl-beta-D-glucopyranoside dihydrochloride
##STR00018##
[0044] Scheme 3, step B: Add hydrogen chloride (4.0 M solution in
1,4-dioxane, 1.5 mL, 5.8 mmol) to a solution of tert-butyl
2-{(3E)-4-[3-methyl-4-({5-(propan-2-yl)-3-[(2,3,4,6-tetra-O-acetyl-beta-D-
-glucopyranosyl)oxy]-1H-pyrazol-4-yl}methyl)phenyl]but-3-en-1yl}-2,9-diaza-
spiro[5.5]undecane-9-carboxylate (500 mg, 0.58 mmol) in
dichloromethane (20 mL). After 2 hours at room temperature, remove
the solvent under reduced pressure to provide the title compound as
solid (480 mg, 0.57 mmol). MS (m/z): 767.4 (M+1).
Preparation 11
tert-butyl
[2-(2-{(3E)-4-[3-methyl-4-({5-(propan-2-yl)-3-[(2,3,4,6-tetra-O-
-acetyl-beta-D-glucopyranosyl)oxy]-1H-pyrazol-4-yl}methyl)phenyl]but-3-en--
1-yl}-2,9-diazaspiro[5.5]undec-9-yl)-2-oxoethyl]carbamate
##STR00019##
[0046] Scheme 3, step C: Add
O-(7-azabenzotriazol-1-yl)-N,N,N',N'tetramethyluronium
hexafluorophosphate (HATU, 543 mg, 1.43 mmol) to a solution of
4-{4-[(1E)-4-(2,9-diazaspiro[5.5]undec-2-yl)but-1-en-1-yl]-2-methylbenzyl-
}-5-(propan-2-yl)-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyyl-beta-D-glucopyranoside dihydrochloride (1.0
g, 1.19 mmol), N-alpha-T-Boc-glycine (317 mg, 1.79 mmol), and
diisopropylethylamine (769 mg, 5.95 mmol) in dichloromethane (10
mL). Stir the mixture at room temperature for 2.0 hours. Remove the
solvent under the reduced pressure. Purify the resulting residue by
flash chromatography (silica gel, gradient ethyl
acetate/dichloromethane from 30-85% over 20 mM, 40 g column) to
provide the title compound (1.0 g, 1.08 mmol). MS (m/z): 924.4
(M+1), 922.4 (M-1).
Alternative synthesis of tert-butyl
[2-(2-{(3E)-4-[3-methyl-4-({5-(propan-2-yl)-3-[(2,3,4,6-tetra-O-acetyl-be-
ta-D-glucopyranosyl)oxy]-1H-pyrazol-4-yl}methyl)phenyl]but-3-en-1-yl}-2,9--
diazaspiro[5.5]undec-9-yl)-2-oxoethyl]carbamate
[0047] Scheme 3, step C: Note that the starting material lot used
for this transformation is about 64% pure (measured using HPLC UV
absorbance). A flask is charged with tert-butyl
2-{(3E)-4-[3-methyl-4-({5-(propan-2-yl)-3-[(2,3,4,6-tetra-O-acetyl-beta-D-
-glucopyranosyl)oxy]-1H-pyrazol-4-yl}methyl)phenyl]but-3-en-1-yl}-2,9-diaz-
aspiro[5.5]undecane-9-carboxylate (from preparation 9; 6.70 g, 7.73
mmoles) dichloromethane (40.00 mL) and hydrogen chloride (4M in
dioxane; 9.66 mL, 38.64 mmoles). The mixture is stirred at ambient
temperature for 3 hours. The solvent is removed under reduced
pressure to provide
4-[4-{(1E)-4-(2,9-diazaspiro[5.5]undec-2-yl)but-1-en-1-yl]-2-methylbenzyl-
}-5-(propan-2-yl)-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyyl-beta-D-glucopyranoside dihydrochloride. To
this is added dichloromethane (32.50 mL) and then triethylamine
(5.39 mL, 38.70 mmoles). This mixture is stirred to give a milky
suspension. A second flask is charged with N-alpha-T-BOC-glycine
(1.63 g, 9.29 mmoles), then dichloromethane (32.50 mL), and then
1,1'-carbonyldiimidazole (1.57 g, 9.67 mmoles). The mixture is
stirred for 15 minutes. The glycine mixture is then added to the
amine mixture, and the reaction is stirred rapidly for 15 minutes.
Saturated aqueous NaHCO.sub.3 (25 mL) is added and the mixture
stirred for 5 minutes. The phases are separated and the aqueous
phase is extracted with dichloromethane (25 mL). The organic phases
are dried over Na.sub.2SO.sub.4, filtered, and concentrated. The
residue is purified by flash column chromatography eluting with 40
to 100% ethyl acetate in iso-hexane, then further purified by
reverse phase (C18) flash column chromatography eluting with 10 mM
ammonium bicarbonate in water and acetonitrile to provide the title
compound (3 g, 42% yield). Mass spectrum (m/z): 924.4 (M+1).
Preparation 12
4-(4-{(1E)-4-[9-(aminoacetyl)-2,9-diazaspiro[5.5]undec-2-yl]but-1-en-1-yl]-
-2-methylbenzyl}-5-(propan-2-yl)-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyyl-beta-D-glucopyranoside dihydrochloride
##STR00020##
[0049] Scheme 3, step D: Add hydrogen chloride (4.0 M solution in
1,4-dioxane; 1.35 mL, 5.4 mmol) to a solution of tert-butyl
[2-(2-{(3E)-4-{3-methyl-4-({5-(propan-2-yl)-3-[(2,3,4,6-tetra-O-acetyl-be-
ta-D-glucopyranosyl)oxy]-1H-pyrazol-4-yl}methyl)phenyl]but-3-en-1-yl}-2,9--
diazaspiro[5.5]undec-9-yl)-2-oxoethyl]carbamate (1.0 g, 1.08 mmol)
in dichloromethane (10 mL). After 2 hours at room temperature,
concentrate to remove the solvent under reduced pressure to provide
the title compound as a solid (950 mg, 1.06 mmol). MS (m/z): 824.4
(M+1).
Alternative synthesis of
4-(4-{(1E)-4-[9-(aminoacetyl)-2,9-diazaspiro[5.5]undec-2-yl]but-1-en-1-yl-
]-2-methylbenzyl}-5-(propan-2-yl)-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyyl-beta-D-glucopyranoside dihydrochloride
[0050] Scheme 3, step D: A mixture of tert-butyl
[2-(2-{(3E)-4-[3-methyl-4-({5-(propan-2-yl)-3-[(2,3,4,6-tetra-O-acetyl-be-
ta-D-glucopyranosyl)oxy]-1H-pyrazol-4-yl}methyl)phenyl]but-3-en-1-yl}-2,9--
diazaspiro[5.5]undec-9-yl)-2-oxoethyl]carbamate (12.10 g, 13.09
mmoles), dichloromethane (121.00 mL), and hydrogen chloride (4M in
1,4-dioxane; 16.37 mL, 65.47 mmoles) is stirred at ambient
temperature for 4 hours. The mixture is concentrated under reduced
pressure to provide the title compound (11.70 g; 99.6% yield). Mass
spectrum (m/z): 824 (M+1 for free base).
EXAMPLE 1
1-(2-{2-[(3E)-4-(4-{[3-(beta-D-glucopyranosyloxy)-5-(propan-2-yl)-1H-pyraz-
ol-4-yl]methyl}-3-methylphenyl)but-3-en-1-yl]-2,9-diazaspiro[5.5]undec-9-y-
l}-2-oxoethyl)-3-(2-methylpropyl)urea
##STR00021##
[0052] Scheme 3, step E: Add isobutyl isocyanate (77 mg, 0.78 mmol)
to a solution of
4-(4-{(1E)-4-[9-(aminoacetyl)-2,9-diazaspiro[5.5]undec-2-yl]but-1-en-1-yl-
]-2-methylbenzyl}-5-(propan-2-yl)-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyyl-beta-D-glucopyranoside dihydrochloride (350
mg, 0.39 mmol) and diisopropylethylamine (302 mg, 2.34 mmol) in
dichloromethane (1.0 mL). After 30 min at room temperature,
concentrate to dryness under the reduced pressure. Treat the
residue with 2.0 M NH.sub.3/methanol (1.0 mL) for 3 hours at room
temperature, remove the solvent under the reduced pressure. Purify
the resulting residue by preparative HPLC method: high pH, 29% B
for 4 min, 29-44% B for 5 min@85 mL/min using a 30.times.75 mm, 5
um C18XBridge ODB column, solvent A--H.sub.2O w
NH.sub.4HCO.sub.3@pH 10, solvent B--acetonitrile to yield the title
compound as solid (134 mg, 0.18 mmol). MS (m/z): 755.2 (M+1), 753.2
(M-1).
[0053] .sup.1H NMR (400.31 MHz, CD.sub.3OD): 7.10 (d, J=1.3 Hz,
1H), 7.04 (dd,J=1.3, 8.0 Hz, 1H), 6.87 (d, J=8.0 Hz, 1H), 6.36 (d,
J=15.6 Hz, 1H), 6.16 (dt, J=15.6, 6.5 Hz, 1H), 5.02 (m, 1H), 3.96
(d, J=16.4 Hz, 1H), 3.93 (d, J=16.4 Hz, 1H), 3.80 (d, J=12.4 Hz
1H), 3.71 (d, J=17.0 Hz, 1H), 3.68 (d, J=17.0 Hz, 1H), 3.64 (m,
1H), 3.60-3.43 (m, 2H), 3.41-3.31 (m, 6H), 2.93 (d, J=6.8 Hz, 2H),
2.80 (m, 1H), 2.47-2.22 (m, 8H), 2.30 (s, 3H), 1.71 (m, 1H),
1.66-1.32 (m, 8H), 1.11 (d, J=7.0 Hz, 3H), 1.1 (d, J=7.0 Hz, 3H),
0.89 (d, J=6.7 Hz, 6H).
Alternative Synthesis of
1-(2-{2-[(3E)-4-(4-{[3-(beta-D-glucopyranosyloxy)-5-(propan-2-yl)-1H-pyra-
zol-4-yl]methyl}-3-methylphenyl)but-3-en-1-yl]-2,9-diazaspiro[5.5]undec-9--
yl}-2-oxoethyl)-3-(2-methylpropyl)urea
[0054] Scheme 3, step E: A mixture of
4-(4-{(1E)-4-[9-(aminoacetyl)-2,9-diazaspiro[5.5]undec-2-yl]but-1-en-1-yl-
]-2-methylbenzyl}-5-(propan-2-yl)-1H-pyrazol-3-yl
2,3,4,6-tetra-O-acetyyl-beta-D-glucopyranoside dihydrochloride
(11.70 g; 13.04 mmoles) in dichloromethane (33 mL) and
diisopropylethylamine (13.65 mL; 78.27 mmoles) is stirred for 10
minutes to dissolve. The mixture is cooled to 5.degree. C. and
isobutyl isocyanate (2.59 g, 26.09 mmoles) is added dropwise over 5
minutes. When the addition is complete the cooling bath was
removed. After 15 minutes, the mixture is concentrated under
reduced pressure. The residue is treated with ammonia (2M in
methanol; 35.00 mL, 70.00 mmoles) and stirred at ambient
temperature for 1.5 hours. Further ammonia (2M in methanol; 35.00
mL, 70.00 mmoles) is added and the reaction is stirred overnight.
The mixture is warmed to 40.degree. C. and ammonia (2M in methanol;
35.00 mL, 70.00 mmoles) is added. The mixture is stirred for 160
minutes, and then concentrated. The residue is purified by reverse
phase (C18) flash column chromatography eluting with 20:80 to 80:20
10 mM ammonium bicarbonate in water:acetonitrile to give the title
compound (6.686 g, 68% yield). Mass spectrum (m/z): 755.5
(M+1).
Sodium-Dependent Glucose Transporter 1 (SGLT1) and SGLT2 Assays
[0055] The cDNA encoding human SGLT1 (slc5a1, NM.sub.--000343),
human SGLT2 (slc5a2, NM.sub.--003041) and mouse SGLT1 (slc5a1,
NM.sub.--019810.4) are purchased from Openbiosystems, Invitrogen
and Openbiosystems, respectively. The cDNA is cloned into pcDNA3.1+
for mammalian expression and is stably transfected into Chinese
hamster ovary (CHO)-K1 cells using standard mammalian transfection
procedures. An SGLT-expressing sub-clone of each over-expressing
cell line is selected based on resistance to neomycin (Geneticin,
Invitrogen) and activity in the
.sup.14C-.alpha.-methyl-D-glucopyranoside (.sup.14C-AMG) uptake
assay (see below). Stable SGLT-expressing cells are maintained
using standard cell culture techniques.
[0056] The SGLT activity is measured as sodium-dependent
.sup.14C-AMG uptake in the above cell lines described as follows.
One hundred .mu.L of culture medium containing 30,000 cells are
seeded to each well of a 96-well BioCoat poly-D-lysine plate
(Becton Dickson) and cultured at 37.degree. C. overnight. The
culture medium is aspirated and cells are washed twice with 200
.mu.L of Reaction Buffer (140 mM NaCl, 2 mM KCl, 1 mM CaCl.sub.2,
MgCl.sub.2, and 14 mM N-2-hydroethylpiperrazine-N'-2-ethanesulfonic
acid (Hepes), pH 7.5). The excess buffer is tapped out onto paper
towels. Thirty-five .mu.L of Reaction Buffer are added to each
well. Five .mu.L of a 10% dimethylsufoxide (DMSO) in Reaction
Buffer containing varying concentrations of test compound or no
compound as a control, is dispensed into the each well. The
reaction is initiated by adding 10 .mu.L of .sup.14C-AMG in
Reaction Buffer to make a final concentration of 4 .mu.M. The plate
is incubated at 37.degree. C. for 125 minutes. The reaction is
terminated by aspirating off Reaction Buffer and then washed three
times with 200 .mu.L of ice cold Reaction Buffer. Manual aspiration
is applied to ensure the complete removal of Reaction Buffer. Ten
.mu.L of 0.1 N NaOH is added to each well and then 100 .mu.L of
Supermix scintillation cocktail (PerkinElmer) is added. After
mixing, the scintillation signal in the plate is counted in a
MicroBeta (PerkinElmer). A ten-dose response curve is fitted to an
empirical four-parameter model using ActivityBase (ID Business
Solution) to determine the inhibitor concentration at half-maximal
inhibition (IC.sub.50).
TABLE-US-00001 TABLE 1 In vitro potency of Example 1 against SGLT1
and SGLT2 Test Human SGLT1 Human SGLT2 Mouse SGLT1 Compound
IC.sub.50, nM IC.sub.50, nM IC.sub.50, nM Example 1 17.3 .+-. 16.2
(n = 5) 171 .+-. 26 (n = 5) 7.3 .+-. 2.9 (n = 5)
[0057] The data in table 1 demonstrate that the compound of Example
1 inhibits human and mouse SGLT1 in vitro, and is more potent at
human and mouse SGLT1 than at human SGLT2 in vitro.
Glucose Lowering Effects in Oral Glucose Tolerance Test (OGTT)
[0058] The test compound is formulated by adding a vehicle of 1%
hydroxyethylcellulose, 0.25% Tween.RTM. 80 w/antifoam 0.05% to
preweighed test compound to make a lmg/ml solution. The mixture is
probe sonicated for approximately 1 minute. A stir bar is added,
and the resulting suspension is stirred continuously throughout
dosing.
[0059] Single housed C57B1/6 mice are weighed and body weights used
to determine study groups (n=5), within a working range of 26-30 g.
After grouping, all mice are fasted overnight by removing access to
food, late afternoon before test day. At the same time, two groups
of mice are orally gavaged with 10 ml/kg test compound preparation
or vehicle. Animals are dosed thirty seconds apart. These mice are
used to demonstrate the compound's effects in an OGTT, 18 hours
later. The following morning, the remaining mice are weighed and
orally gavaged using the same protocol as the previous day.
[0060] At five, eight and eighteen hours after each respective
compound treatment is started, a baseline blood sample is taken for
measuring glucose (from the first animal, via tail snip). The
animal is then immediately given an oral dose of 50% dextrose
(Hospira.RTM.) at 3 g/kg. Blood samples are taken for glucose,
exactly thirty seconds apart, by tail vein so that blood is
collected in each animal at 20, 40, 60 and 120 minutes after the
dextrose dose.
TABLE-US-00002 TABLE 2 Glucose lowering effects in OGTT. Oral
Glucose Tolerance Test Results Mean .+-. SE Example 1 Example 1
Example 1 Vehicle 10 mg/kg Vehicle 10 mg/kg Vehicle 10 mg/kg @ 5
hrs @ 5 hr @ 8 hrs @ 8 hrs @ 18 hrs @ 18 hrs post Dose post Dose
post Dose post Dose post Dose post Dose Glucose (mg/dl) 0 Minute
74.5 .+-. 1.54 75.9 .+-. 3.21 75.0 .+-. 6.48 75.5 .+-. 4.42 69.7
.+-. 3.42 78.7 .+-. 8.81 20 Minute 232.3 .+-. 13.51 124.7 .+-.
7.66** 296.9 .+-. 26.4 142.4 .+-. 7 4** 282.3 .+-. 12.8 196.0 .+-.
16.6** 40 Minute 189.5 .+-. 12.4 139.2 .+-. 7.69** 240.0 .+-. 15.1
168.1 .+-. 8.37** 241.6 .+-. 17.9 216.1 .+-. 15.0 60 Minute 176.5
.+-. 14.8 133.2 .+-. 6.24** 183.1 .+-. 12.9 171.2 .+-. 10.1 163.3
.+-. 9.77 188.7 .+-. 13.8 120 Minute 104.1 .+-. 8.57 108.7 .+-.
4.93 109.7 .+-. 3.89 113.6 .+-. 4.25 102.5 .+-. 3.57 110.3 .+-.
5.74 Baseline 6475 .+-. 168 2815 .+-. 194** 8819 .+-. 668 4147 .+-.
456** 8626 .+-. 565 6194 .+-. 406* Adjusted AUC Glucose 232.3 .+-.
13.5 140.2 .+-. 7.48** 296.9 .+-. 26.4 175.1 .+-. 9 3** 283.2 .+-.
13.1 216.1 .+-. 15.0* Cmax Time (minutes) Glucose .sup. 20 .+-. 0.0
56 .+-. 16 .sup. 20 .+-. 0.0 52 .+-. 4.9** .sup. 24 .+-. 4.0 .sup.
44 .+-. 4.0* Tmax 1 way ANOVA/Dunnett's *p < 0.05, **p < 0.01
compared to vehicle
[0061] As shown in table 1, the compound of example 1 delivers a
decrease in the glucose excursion when an oral bolus of 50%
dextrose (Hospira.RTM.) is given to a normal glycemic C57B1/6 mouse
five, eight or eighteen hours after administration. Example 1 also
demonstrates a dose dependent decrease in baseline adjusted glucose
area under the curve (AUC) during all three OGTTs. In addition,
example 1 decreases the average maximum concentration of plasma
glucose (Cmax) during all three OGTTs while increasing the average
time that it takes for glucose to reach maximum concentration
(Tmax).
* * * * *