U.S. patent application number 16/716433 was filed with the patent office on 2020-04-16 for crystalline forms of a prolyl hydroxylase inhibitor.
The applicant listed for this patent is FibroGen, Inc.. Invention is credited to Michael P. Arend, Michael John Martinelli, Jung Min Park, Michael D. Thompson, Claudia Witschi, David A. Yeowell.
Application Number | 20200115344 16/716433 |
Document ID | / |
Family ID | 48857025 |
Filed Date | 2020-04-16 |
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United States Patent
Application |
20200115344 |
Kind Code |
A1 |
Witschi; Claudia ; et
al. |
April 16, 2020 |
CRYSTALLINE FORMS OF A PROLYL HYDROXYLASE INHIBITOR
Abstract
The present disclosure relates to crystalline solid forms of
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid, the process of preparing the forms, and pharmaceutical
compositions and methods of use thereof.
Inventors: |
Witschi; Claudia; (San
Francisco, CA) ; Park; Jung Min; (San Francisco,
CA) ; Thompson; Michael D.; (Redwood City, CA)
; Martinelli; Michael John; (San Francisco, CA) ;
Yeowell; David A.; (Chapel Hill, NC) ; Arend; Michael
P.; (Foster City, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
FibroGen, Inc. |
San Francisco |
CA |
US |
|
|
Family ID: |
48857025 |
Appl. No.: |
16/716433 |
Filed: |
December 16, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16158242 |
Oct 11, 2018 |
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16716433 |
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|
15462688 |
Mar 17, 2017 |
10118897 |
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16158242 |
|
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|
14805345 |
Jul 21, 2015 |
9617218 |
|
|
15462688 |
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13942443 |
Jul 15, 2013 |
9115085 |
|
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14805345 |
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61832566 |
Jun 7, 2013 |
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61768297 |
Feb 22, 2013 |
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61672191 |
Jul 16, 2012 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/47 20130101;
A61P 13/12 20180101; A61P 33/00 20180101; C07B 2200/13 20130101;
A61P 1/16 20180101; A61P 25/28 20180101; A61P 11/00 20180101; C07D
217/26 20130101; A61P 17/00 20180101; A61P 17/02 20180101; A61P
25/00 20180101; A61P 25/08 20180101; A61P 1/00 20180101; A61P 1/04
20180101; A61P 31/18 20180101; A61P 9/00 20180101; A61P 9/04
20180101; A61P 7/06 20180101; A61P 3/10 20180101; A61P 27/02
20180101; A61P 43/00 20180101; A61P 31/04 20180101; A61P 35/00
20180101; A61P 9/10 20180101; A61P 9/12 20180101; A61P 39/00
20180101; A61P 29/00 20180101 |
International
Class: |
C07D 217/26 20060101
C07D217/26 |
Claims
1. A process for preparing crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid (Compound A), comprising contacting Compound A, or a salt
thereof, with acetic acid for a time sufficient to provide a
crystalline Compound A wherein at least about 95% of the
crystalline Compound A is Form A.
2. The process of claim 1, wherein the process further comprises,
following said contacting step, isolating the crystalline Compound
A Form A.
3. The process of claim 2, wherein the isolating comprises the
steps of filtering, washing and drying the crystalline Compound A
Form A.
4. The process of claim 1, wherein the contacting comprises adding
the acetic acid to Compound A at a temperature of from about
10.degree. C. to about 90.degree. C.
5. The process of claim 4, wherein the acetic acid is added to the
Compound A as an aqueous solution, slowly with stirring.
6. The process of claim 1, wherein prior to contacting Compound A
with the acetic acid, an aqueous solution of sodium hydroxide is
added slowly to a stirred suspension of Compound A in water at
temperature of from about 10.degree. C. to about 90.degree. C.
7. The process of claim 1, wherein the Compound A Form A is
characterized by a differential scanning calorimetry (DSC) curve
that comprises an endotherm at about 223.degree. C.
8. The process of claim 7, wherein the DSC curve is substantially
as shown in FIG. 2.
9. The process of claim 1, wherein the Compound A Form A is
characterized by an X-ray powder diffractogram comprising a peak at
8.5.degree.2.theta..+-.0.2.degree.2.theta..
10. The process of claim 1, wherein the Compound A Form A is
characterized by an X-ray powder diffractogram comprising a peak at
16.2.degree.2.theta..+-.0.2.degree.2.theta..
11. The process of claim 1, wherein the Compound A Form A is
characterized by an X-ray powder diffractogram comprising a peak at
27.4.degree.2.theta..+-.0.2.degree.2.theta..
12. The process of claim 1, wherein the Compound A Form A is
characterized by an X-ray powder diffractogram comprising peaks at
8.5, 16.2, and 27.4.degree.2.theta..+-.0.2.degree.2.theta..
13. The process of claim 12, wherein the X-ray powder diffractogram
further comprises peaks at 12.8, 21.6, and
22.9.degree.2.theta..+-.0.2.degree.2.theta..
14. The process of claim 1, wherein the Compound A Form A is
characterized by an X-ray powder diffractogram substantially as
shown in FIG. 1.
15. The process of claim 1, wherein at least about 99% of the
crystalline Compound A produced by the process is Compound A Form
A.
16. The process of claim 1, wherein at least about 99.5% of the
crystalline Compound A produced by the process is Compound A Form
A.
17. The process of claim 1, wherein at least about 99.9% of the
crystalline Compound A produced by the process is Compound A Form
A.
18. The process of claim 1, wherein at least about 99.99% of the
crystalline Compound A produced by the process is Compound A Form
A.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 16/158,242, filed Oct. 11, 2018, which is a continuation of
U.S. application Ser. No. 15/462,688, filed Mar. 17, 2017, now U.S.
Pat. No. 10,118,897, which is a continuation of U.S. application
Ser. No. 14/805,345, filed Jul. 21, 2015, now U.S. Pat. No.
9,617,218, which is a continuation of U.S. application Ser. No.
13/942,443, filed Jul. 15, 2013, now U.S. Pat. No. 9,115,085, which
application claims the benefit under 35 U.S.C. .sctn. 119(e) to
U.S. Provisional Application No. 61/672,191, filed Jul. 16, 2012,
U.S. Provisional Application No. 61/768,297, filed Feb. 22, 2013,
and U.S. Provisional Application No. 61/832,566, filed Jun. 7,
2013, all of which are hereby incorporated by reference in their
entireties.
FIELD
[0002] The present disclosure relates to crystalline solid forms of
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid, the process of preparing the forms, and pharmaceutical
compositions and methods of use thereof.
STATE OF THE ART
[0003] A compound can exist in one or more crystalline forms.
Crystalline forms of a drug substance can have different physical
properties, including melting point, solubility, dissolution rate,
optical and mechanical properties, vapor pressure, hygroscopicity,
particle shape, density, and flowability. These properties can have
a direct effect on the ability to process and/or manufacture a
compound as a drug product. Crystalline forms can also exhibit
different stabilities and bioavailability. The most stable
crystalline form of a drug product is often chosen during drug
development based on the minimal potential for conversion to
another crystalline form and on its greater chemical stability. To
ensure the quality, safety, and efficacy of a drug product, it is
important to choose a crystalline form that is stable, is
manufactured reproducibly, and has favorable physicochemical
properties.
[0004]
[(4-Hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acet-
ic acid (hereinafter, Compound A) is a potent inhibitor of hypoxia
inducible factor (HIF) prolyl hydroxylase, as described in U.S.
Pat. No. 7,323,475. HIF prolyl hydroxylase inhibitors are useful
for increasing the stability and/or activity of HIF, and useful
for, inter alia, treating and preventing disorders associated with
HIF, including anemia, ischemia, and hypoxia.
SUMMARY
[0005] The present disclosure fulfills these needs and others by
providing crystalline forms of Compound A, salts, and solvates. The
present disclosure also provides an amorphous form of Compound A.
The present disclosure also provides pharmaceutical compositions
comprising amorphous or one or more crystalline forms of Compound
A. The disclosure also provides processes for making the amorphous
and crystalline solid forms and methods for using them to treat,
and prevent HIF-associated disorders including conditions involving
anemia, ischemia, and hypoxia.
[0006] Thus, one embodiment provided is crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid (Compound A Form A) characterized by an X-ray powder
diffractogram comprising the following peaks: 8.5, 16.2, and
27.4.degree.2.theta..+-.0.2.degree.2.theta..
[0007] Another embodiment provided is crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid hemihydrate (Compound A Form B) characterized by an X-ray
powder diffractogram comprising the following peaks: 4.2, 8.3, and
16.6.degree.2.theta..+-.0.2.degree.2.theta..
[0008] Yet another embodiment provided is crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid hexafluoropropan-2-ol solvate (Compound A Form C)
characterized by an X-ray powder diffractogram comprising the
following peaks: 4.5, 13.7, and
16.4.degree.2.theta..+-.0.2.degree.2.theta..
[0009] Yet another embodiment provided is crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid DMSO:water solvate (Compound A Form D) characterized by an
X-ray powder diffractogram comprising the following peaks: 8.4,
8.5, and 16.8.degree.2.theta..+-.0.2.degree.2.theta..
[0010] Yet another embodiment provided is crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid sodium salt (Compound A sodium salt) characterized by an X-ray
powder diffractogram comprising the following peaks: 5.3, 16.0, and
21.6.degree.2.theta..+-.0.2.degree.2.theta..
[0011] Yet another embodiment provided is crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid L-arginine salt (Compound A L-arginine salt) characterized by
an X-ray powder diffractogram comprising the following peaks: 20.8,
21.8, and 25.4.degree.2.theta..+-.0.2.degree.2.theta..
[0012] Yet another embodiment provided is crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid L-lysine salt (Compound A L-lysine salt) characterized by an
X-ray powder diffractogram comprising the following peaks: 19.8,
20.7, and 21.2.degree.2.theta..+-.0.2.degree.2.theta..
[0013] Yet another embodiment provided is crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid ethanolamine salt (Compound A ethanolamine salt) characterized
by an X-ray powder diffractogram comprising the following peaks:
21.8, 22.7, and 27.1.degree.2.theta..+-.0.2.degree.2.theta..
[0014] Yet another embodiment provided is crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid diethanolamine salt (Compound A diethanolamine salt)
characterized by an X-ray powder diffractogram comprising the
following peaks: 16.9, 23.7, and
25.0.degree.2.theta..+-.0.2.degree.2.theta..
[0015] Yet another embodiment provided is crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid tromethamine salt (Compound A tromethamine salt) characterized
by an X-ray powder diffractogram comprising the following peaks:
10.1, 14.2, and 21.1.degree.2.theta..+-.0.2.degree.2.theta..
[0016] Yet another embodiment provided is amorphous
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid (amorphous Compound A).
[0017] Yet another embodiment provided is substantially amorphous
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid potassium salt (Compound A potassium salt).
[0018] Still another embodiment provided is directed to a
pharmaceutical composition comprising a crystalline or amorphous
form of Compound A, or a salt thereof, and a pharmaceutically
acceptable excipient.
[0019] Additionally, the disclosure provides in one embodiment a
method for treating, pretreating, or delaying onset or progression
of a condition mediated at least in part by hypoxia inducible
factor (HIF). The method comprises administering to a patient in
need thereof a therapeutically effective amount of a compound
selected from the group consisting of: Compound A Form A, Compound
A Form B, Compound A Form C, Compound A Form D, Compound A sodium
salt, Compound A L-arginine salt, Compound A L-lysine salt,
Compound A ethanolamine salt, Compound A diethanolamine salt,
Compound A tromethamine salt, amorphous Compound A, and Compound A
potassium salt, as described generally above.
[0020] Also provided is a method for treating, pretreating, or
delaying onset or progression of a condition mediated at least in
part by erythropoietin (EPO), comprising administering to a patient
in need thereof, a therapeutically effective amount of a compound
selected from the group consisting of: Compound A Form A, Compound
A Form B, Compound A Form C, Compound A Form D, Compound A sodium
salt, Compound A L-arginine salt, Compound A L-lysine salt,
Compound A ethanolamine salt, Compound A diethanolamine salt,
Compound A tromethamine salt, amorphous Compound A, and Compound A
potassium salt, as described generally above.
[0021] Also provided is a method for treating, pretreating, or
delaying onset or progression of anemia, comprising administering
to a patient in need thereof, a therapeutically effective amount of
a compound selected from the group consisting of: Compound A Form
A, Compound A Form B, Compound A Form C, Compound A Form D,
Compound A sodium salt, Compound A L-arginine salt, Compound A
L-lysine salt, Compound A ethanolamine salt, Compound A
diethanolamine salt, Compound A tromethamine salt, amorphous
Compound A, and Compound A potassium salt, as described generally
above.
[0022] Also provided is a method of inhibiting the activity of a
HIF hydroxylase enzyme, the method comprising bringing into contact
the HIF hydroxylase enzyme and a therapeutically effective amount
of a compound selected from the group consisting of: Compound A
Form A, Compound A Form B, Compound A Form C, Compound A Form D,
Compound A sodium salt, Compound A L-arginine salt, Compound A
L-lysine salt, Compound A ethanolamine salt, Compound A
diethanolamine salt, Compound A tromethamine salt, amorphous
Compound A, and Compound A potassium salt, as described generally
above.
BRIEF DESCRIPTION OF THE DRAWINGS
[0023] FIG. 1 is an X-ray powder diffraction pattern of Compound A
Form A.
[0024] FIG. 2 is a differential scanning calorimetry (DSC) curve of
Compound A Form A.
[0025] FIG. 3 is an X-ray powder diffraction pattern of Compound A
Form B (bottom) plotted with an X-ray powder diffraction pattern of
Compound A Form A (top).
[0026] FIG. 4 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A Form B.
[0027] FIG. 5 is an X-ray powder diffraction pattern of Compound A
Form C (bottom) plotted with an X-ray powder diffraction pattern of
Compound A Form A (top).
[0028] FIG. 6 is a differential scanning calorimetry (DSC) curve
(top) and a thermogravimetric analysis (TGA) (bottom) of Compound A
Form C.
[0029] FIG. 7 is an X-ray powder diffraction pattern of Compound A
Form D.
[0030] FIG. 8 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A Form D.
[0031] FIG. 9 is an X-ray powder diffraction pattern of Compound A
sodium salt as isolated (bottom) and at 40.degree. C./75% RH
(top).
[0032] FIG. 10 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A sodium salt.
[0033] FIG. 11 is an X-ray powder diffraction pattern of Compound A
L-arginine salt as isolated (bottom) and at 40.degree. C./75% RH
(top).
[0034] FIG. 12 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A L-arginine salt.
[0035] FIG. 13 is an X-ray powder diffraction pattern of Compound A
L-lysine salt as isolated (bottom) and at 40.degree. C./75% RH
(top).
[0036] FIG. 14 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A L-lysine salt.
[0037] FIG. 15 is an X-ray powder diffraction pattern of Compound A
Form A (bottom), Compound A ethanolamine salt pattern 1 as isolated
(second to bottom), Compound A ethanolamine salt pattern 3 at
40.degree. C./75% RH (middle), Compound A ethanolamine salt pattern
2 as isolated (second to top), and Compound A ethanolamine salt
pattern 2 at 40.degree. C./75% RH (top).
[0038] FIG. 16 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A ethanolamine salt.
[0039] FIG. 17 is an X-ray powder diffraction pattern of Compound A
Form A (bottom), Compound A diethanolamine salt pattern 1 from
acetone (second to bottom), Compound A diethanolamine salt pattern
1 from THF (second to top), and Compound A diethanolamine salt at
40.degree. C./75% RH (pattern 2, top).
[0040] FIG. 18 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A diethanolamine salt.
[0041] FIG. 19 is an X-ray powder diffraction pattern of Compound A
Form A (bottom), and Compound A tromethamine salt as isolated
(middle) and at 40.degree. C./75% RH (top).
[0042] FIG. 20 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A tromethamine salt.
[0043] FIG. 21 is an X-ray powder diffraction pattern of Compound A
potassium salt as isolated (bottom) and at 40.degree. C./75% RH
(top).
[0044] FIG. 22 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A potassium salt.
[0045] FIG. 23 is an X-ray powder diffraction pattern of amorphous
Compound A.
[0046] FIG. 24 is the thermogravimetric analysis (TGA) of Compound
A Form A.
[0047] FIG. 25 is an X-ray powder diffraction pattern of Compound A
Form A (bottom), and Compound A hydrochloric acid salt as isolated
(middle) and at 40.degree. C./75% RH (top).
[0048] FIG. 26 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A hydrochloric acid salt.
[0049] FIG. 27 is an X-ray powder diffraction pattern of Compound A
Form A (bottom), and Compound A sulfuric acid salt as isolated
(middle) and at 40.degree. C./75% RH (top).
[0050] FIG. 28 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A sulfuric acid salt.
[0051] FIG. 29 is an X-ray powder diffraction pattern of Compound A
Form A (bottom), Compound A methanesulfonic acid salt pattern 1 as
isolated (second to bottom) and at 40.degree. C./75% RH (middle),
and Compound A methanesulfonic acid salt pattern 2 as isolated
(second to top) and at 40.degree. C./75% RH (top).
[0052] FIG. 30 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A methanesulfonic acid salt.
[0053] FIG. 31 is an X-ray powder diffraction pattern of Compound A
Form A (bottom), Compound A bis triethylamine salt as isolated
(middle) and Compound A bis triethylamine salt at 40.degree. C./75%
RH (top).
[0054] FIG. 32 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A bis triethylamine salt.
[0055] FIG. 33 is an X-ray powder diffraction pattern of Compound A
Form A (bottom), and Compound A hemi calcium salt (second crop) at
40.degree. C./75% RH (top).
[0056] FIG. 34 is a thermogravimetric analysis (TGA) (top) and a
differential scanning calorimetry (DSC) curve (bottom) of Compound
A hemi calcium salt.
[0057] FIG. 35 is an X-ray powder diffraction pattern of Compound A
Form A (bottom), and Compound A hemi magnesium salt (second crop)
at 40.degree. C./75% RH (top).
[0058] FIG. 36 is a differential scanning calorimetry (DSC) curve
of Compound A hemi magnesium salt.
[0059] FIG. 37 is the molecular configuration of Compound A Form
A.
DETAILED DESCRIPTION
[0060] The compound
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid (Compound A) is a potent inhibitor of hypoxia inducible factor
(HIF) prolyl hydroxylase and has the following formula:
##STR00001##
[0061] The present disclosure provides crystalline forms of
Compound A, salts of Compound A, and solvates of Compound A. The
present disclosure also provides an amorphous form of Compound A.
The present disclosure also provides pharmaceutical compositions
comprising amorphous or crystalline forms of Compound A. The
disclosure also provides processes for making the amorphous and
crystalline solid forms and methods for using them to treat, and
prevent HIF-associated disorders including conditions involving
anemia, ischemia, and hypoxia.
[0062] Prior to discussing in further detail, the following terms
will be defined.
1. Definitions
[0063] As used herein, the following terms have the following
meanings.
[0064] The singular forms "a," "an," and "the" and the like include
plural referents unless the context clearly dictates otherwise.
Thus, for example, reference to "a compound" includes both a single
compound and a plurality of different compounds.
[0065] The term "about" when used before a numerical designation,
e.g., temperature, time, amount, and concentration, including a
range, indicates approximations which may vary by .+-.10%, .+-.5%
or .+-.1%.
[0066] The term "solvate" refers to a complex formed by the
combining of Compound A and a solvent.
[0067] The terms "substantially amorphous" and "mostly amorphous"
refer to amorphous Compound A where a small amount of crystalline
Compound A may be present. In some embodiments, the amount of
crystalline Compound A is less than about 10%, or less than about
5%, or less than about 2%, or less than about 1%, or less than
about 0.2%, or less than about 0.1%.
[0068] "Administration" refers to introducing an agent into a
patient. A therapeutic amount can be administered, which can be
determined by the treating physician or the like. An oral route of
administration is preferred for the crystalline forms of Compound A
described herein. The related terms and phrases "administering" and
"administration of", when used in connection with a compound or
pharmaceutical composition (and grammatical equivalents) refer both
to direct administration, which may be administration to a patient
by a medical professional or by self-administration by the patient,
and/or to indirect administration, which may be the act of
prescribing a drug. For example, a physician who instructs a
patient to self-administer a drug and/or provides a patient with a
prescription for a drug is administering the drug to the patient.
In any event, administration entails delivery of the drug to the
patient.
[0069] "Excipient" as used herein means an inert or inactive
substance used in the production of pharmaceutical products,
including without limitation any substance used as a binder,
disintegrant, coating, compression/encapsulation aid, cream or
lotion, lubricant, parenteral, sweetener or flavoring,
suspending/gelling agent, or wet granulation agent. Binders
include, e.g., carbopol, povidone, xanthan gum, etc.; coatings
include, e.g., cellulose acetate phthalate, ethylcellulose, gellan
gum, maltodextrin, etc.; compression/encapsulation aids include,
e.g., calcium carbonate, dextrose, fructose, honey, lactose
(anhydrate or monohydrate; optionally in combination with
aspartame, cellulose, or microcrystalline cellulose), starch,
sucrose, etc.; disintegrants include, e.g., croscarmellose sodium,
gellan gum, sodium starch glycolate, etc.; creams and lotions
include, e.g., maltodextrin, carrageenans, etc.; lubricants
include, e.g., magnesium stearate, stearic acid, sodium stearyl
fumarate, etc.; materials for chewable tablets include, e.g.,
dextrose, fructose dc, lactose (monohydrate, optionally in
combination with aspartame or cellulose), etc.; parenterals
include, e.g., mannitol, povidone, etc.; plasticizers include,
e.g., dibutyl sebacate, polyvinylacetate phthalate, etc.;
suspending/gelling agents include, e.g., carrageenan, sodium starch
glycolate, xanthan gum, etc.; sweeteners include, e.g., aspartame,
dextrose, fructose, sorbitol, sucrose, etc.; and wet granulation
agents include, e.g., calcium carbonate, maltodextrin,
microcrystalline cellulose, etc.
[0070] "Therapeutically effective amount" or "therapeutic amount"
refers to an amount of a drug or an agent that when administered to
a patient suffering from a condition, will have the intended
therapeutic effect, e.g., alleviation, amelioration, palliation or
elimination of one or more manifestations of the condition in the
patient. The therapeutically effective amount will vary depending
upon the subject and the condition being treated, the weight and
age of the subject, the severity of the condition, the particular
composition or excipient chosen, the dosing regimen to be followed,
timing of administration, the manner of administration and the
like, all of which can be determined readily by one of ordinary
skill in the art. The full therapeutic effect does not necessarily
occur by administration of one dose, and may occur only after
administration of a series of doses. Thus, a therapeutically
effective amount may be administered in one or more
administrations. For example, and without limitation, a
therapeutically effective amount of an agent, in the context of
treating anemia, refers to an amount of the agent that alleviates,
ameliorates, palliates, or eliminates one or more symptoms of
anemia in the patient.
[0071] "Treatment", "treating", and "treat" are defined as acting
upon a disease, disorder, or condition with an agent to reduce or
ameliorate the harmful or any other undesired effects of the
disease, disorder, or condition and/or its symptoms. Treatment, as
used herein, covers the treatment of a human patient, and includes:
(a) reducing the risk of occurrence of the condition in a patient
determined to be predisposed to the disease but not yet diagnosed
as having the condition, (b) impeding the development of the
condition, and/or (c) relieving the condition, i.e., causing
regression of the condition and/or relieving one or more symptoms
of the condition.
[0072] An "XRPD pattern" is an x-y graph with diffraction angle
(i.e., .degree. 2.theta.) on the x-axis and intensity on the
y-axis. The peaks within this pattern may be used to characterize a
crystalline solid form. As with any data measurement, there is
variability in XRPD data. The data are often represented solely by
the diffraction angle of the peaks rather than including the
intensity of the peaks because peak intensity can be particularly
sensitive to sample preparation (for example, particle size,
moisture content, solvent content, and preferred orientation
effects influence the sensitivity), so samples of the same material
prepared under different conditions may yield slightly different
patterns; this variability is usually greater than the variability
in diffraction angles. Diffraction angle variability may also be
sensitive to sample preparation. Other sources of variability come
from instrument parameters and processing of the raw X-ray data:
different X-ray instruments operate using different parameters and
these may lead to slightly different XRPD patterns from the same
solid form, and similarly different software packages process X-ray
data differently and this also leads to variability. These and
other sources of variability are known to those of ordinary skill
in the pharmaceutical arts. Due to such sources of variability, it
is usual to assign a variability of .+-.0.2.degree.2.theta. to
diffraction angles in XRPD patterns.
2. Solid Forms of Compound A
[0073] As described generally above, the present disclosure
provides solid forms of
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-a-
cetic acid (Compound A).
[0074] Compound A Form A is characterized by its X-ray powder
diffractogram that comprises peaks at 8.5, 16.2, and
27.4.degree.2.theta..+-.0.2.degree.2.theta.. The diffractogram
comprises additional peaks at 12.8, 21.6, and
22.9.degree.2.theta..+-.0.2.degree.2.theta.. Form A also is
characterized by its full X-ray powder diffractogram as
substantially shown in FIG. 1.
[0075] In some embodiments, Form A is characterized by its
differential scanning calorimetry (DSC) curve that comprises an
endotherm at about 223.degree. C. Form A also is characterized by
its full DSC curve as substantially as shown in FIG. 2.
[0076] Compound A Form B is characterized by its X-ray powder
diffractogram that comprises peaks at 4.2, 8.3, and
16.6.degree.2.theta..+-.0.2.degree.2.theta.. The diffractogram
comprises additional peaks at 12.5, 14.1, and
17.4.degree.2.theta..+-.0.2.degree.2.theta.. Form B also is
characterized by its full X-ray powder diffractogram as
substantially shown in FIG. 3.
[0077] In some embodiments, Form B is characterized by its
differential scanning calorimetry (DSC) curve that comprises an
endotherm at about 222.degree. C. Form B also is characterized by
its full DSC curve as substantially as shown in FIG. 4.
[0078] Compound A Form C is characterized by its X-ray powder
diffractogram that comprises peaks at 4.5, 13.7, and
16.4.degree.2.theta..+-.0.2.degree.2.theta.. The diffractogram
comprises additional peaks at 15.4, 15.5, and
20.6.degree.2.theta..+-.0.2.degree.2.theta.. Form C also is
characterized by its full X-ray powder diffractogram as
substantially shown in FIG. 5.
[0079] In some embodiments, Form C is characterized by its
differential scanning calorimetry (DSC) curve that comprises an
endotherm at about 222.degree. C. Form C also is characterized by
its full DSC curve as substantially as shown in FIG. 6.
[0080] Compound A Form D is characterized by its X-ray powder
diffractogram that comprises peaks at 8.4, 8.5, and
16.8.degree.2.theta..+-.0.2.degree.2.theta.. The diffractogram
comprises additional peaks at 4.2, 12.6, and
28.4.degree.2.theta..+-.0.2.degree.2.theta.. Form D also is
characterized by its full X-ray powder diffractogram as
substantially shown in FIG. 7.
[0081] In some embodiments, Form D is characterized by its
differential scanning calorimetry (DSC) curve that comprises an
endotherm at about 222.degree. C. Form D also is characterized by
its full DSC curve as substantially as shown in FIG. 8.
[0082] Compound A sodium salt is characterized by its X-ray powder
diffractogram that comprises peaks at 5.3, 16.0, and
21.6.degree.2.theta..+-.0.2.degree.2.theta.. The diffractogram
comprises additional peaks at 18.7, 19.2, and
24.0.degree.2.theta..+-.0.2.degree.2.theta.. Compound A sodium salt
also is characterized by its full X-ray powder diffractogram as
substantially shown in FIG. 9.
[0083] In some embodiments, Compound A sodium salt is characterized
by its differential scanning calorimetry (DSC) curve that comprises
an endotherm at about 314.degree. C. Compound A sodium salt also is
characterized by its full DSC curve as substantially as shown in
FIG. 10.
[0084] Compound A L-arginine salt is characterized by its X-ray
powder diffractogram that comprises peaks at 20.8, 21.8, and
25.4.degree.2.theta..+-.0.2.degree.2.theta.. The diffractogram
comprises additional peaks at 22.7, 23.4, and
26.4.degree.2.theta..+-.0.2.degree.2.theta.. Compound A L-arginine
salt also is characterized by its full X-ray powder diffractogram
as substantially shown in FIG. 11.
[0085] In some embodiments, Compound A L-arginine salt is
characterized by its differential scanning calorimetry (DSC) curve
that comprises an endotherm at about 210.degree. C. Compound A
L-arginine salt also is characterized by its full DSC curve as
substantially as shown in FIG. 12.
[0086] Compound A L-lysine salt is characterized by its X-ray
powder diffractogram that comprises peaks at 19.8, 20.7, and
21.2.degree.2.theta..+-.0.2.degree.2.theta.. The diffractogram
comprises additional peaks at 10.2, 16.9, and
18.4.degree.2.theta..+-.0.2.degree.2.theta.. Compound A L-lysine
salt also is characterized by its full X-ray powder diffractogram
as substantially shown in FIG. 13.
[0087] In some embodiments, Compound A L-lysine salt is
characterized by its differential scanning calorimetry (DSC) curve
that comprises an endotherm at about 237.degree. C. Compound A
L-lysine salt also is characterized by its full DSC curve as
substantially as shown in FIG. 14.
[0088] Compound A ethanolamine salt is characterized by its X-ray
powder diffractogram that comprises peaks at 21.8, 22.7, and
27.1.degree.2.theta..+-.0.2.degree.2.theta.. The diffractogram
comprises additional peaks at 21.1, 26.2, and
26.6.degree.2.theta..+-.0.2.degree.2.theta.. Compound A
ethanolamine salt also is characterized by its full X-ray powder
diffractogram as substantially shown in FIG. 15.
[0089] In some embodiments, Compound A ethanolamine salt is
characterized by its differential scanning calorimetry (DSC) curve
that comprises an endotherm at about 171.degree. C. Compound A
ethanolamine salt also is characterized by its full DSC curve as
substantially as shown in FIG. 16.
[0090] Compound A diethanolamine salt is characterized by its X-ray
powder diffractogram that comprises peaks at 16.9, 23.7, and
25.0.degree.2.theta..+-.0.2.degree.2.theta.. The diffractogram
comprises additional peaks at 19.6, 22.6, and
26.0.degree.2.theta..+-.0.2.degree.2.theta.. Compound A
diethanolamine salt also is characterized by its full X-ray powder
diffractogram as substantially shown in FIG. 17.
[0091] In some embodiments, Compound A diethanolamine salt is
characterized by its differential scanning calorimetry (DSC) curve
that comprises an endotherm at about 150.degree. C. Compound A
diethanolamine salt also is characterized by its full DSC curve as
substantially as shown in FIG. 18.
[0092] Compound A tromethamine salt is characterized by its X-ray
powder diffractogram that comprises peaks at 10.1, 14.2, and
21.1.degree.2.theta..+-.0.2.degree.2.theta.. The diffractogram
comprises additional peaks at 20.1, 25.7, and
28.4.degree.2.theta..+-.0.2.degree.2.theta.. Compound A
tromethamine salt also is characterized by its full X-ray powder
diffractogram as substantially shown in FIG. 19.
[0093] In some embodiments, Compound A tromethamine salt is
characterized by its differential scanning calorimetry (DSC) curve
that comprises an endotherm at about 176.degree. C. Compound A
tromethamine salt also is characterized by its full DSC curve as
substantially as shown in FIG. 20.
[0094] Also provided is amorphous
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid (amorphous Compound A) and substantially amorphous
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid potassium salt (Compound A potassium salt). The substantially
amorphous Compound A potassium salt has been characterized by a
differential scanning calorimetry (DSC) curve that comprises an
endotherm at about 291.degree. C. (FIG. 22).
[0095] As described in the Examples below, Form A is the most
stable crystalline form among Form B, C, and D of Compound A.
3. Pharmaceutical Compositions, Formulations and Routes of
Administration
[0096] In one aspect, the present disclosure is directed to a
pharmaceutical composition comprising one or more crystalline forms
of
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid (Compound A) having the following structure:
##STR00002##
or a salt thereof, and at least one pharmaceutically acceptable
excipient.
[0097] In one embodiment, the pharmaceutical composition comprises
a compound selected from the group consisting of: Compound A Form
A, Compound A Form B, Compound A Form C, Compound A Form D,
Compound A sodium salt, Compound A L-arginine salt, Compound A
L-lysine salt, Compound A ethanolamine salt, Compound A
diethanolamine salt, Compound A tromethamine salt, amorphous
Compound A, and Compound A potassium salt, as described generally
above, and at least one pharmaceutically acceptable excipient.
[0098] In one embodiment, the pharmaceutical composition comprises
Compound A in Form A. In one embodiment, the pharmaceutical
composition comprises Compound A wherein at least about 85% of
Compound A is in Form A. In one embodiment, the pharmaceutical
composition comprises Compound A wherein at least about 90% of
Compound A is in Form A. In one embodiment, the pharmaceutical
composition comprises Compound A wherein at least about 95% of
Compound A is in Form A. In one embodiment, the pharmaceutical
composition comprises Compound A wherein at least about 99% of
Compound A is in Form A. In one embodiment, the pharmaceutical
composition comprises Compound A wherein at least about 99.5% of
Compound A is in Form A. In one embodiment, the pharmaceutical
composition comprises Compound A wherein at least about 99.9% of
Compound A is in Form A. In one embodiment, the pharmaceutical
composition comprises Compound A wherein at least about 99.99% of
Compound A is in Form A.
[0099] In one embodiment, the pharmaceutical composition further
comprises an additional therapeutic agent selected from the group
consisting of vitamin B12, folic acid, ferrous sulfate, recombinant
human erythropoietin, and an erythropoiesis stimulating agent
(ESA). In another embodiment, the pharmaceutical composition is
formulated for oral delivery. In another embodiment, the
pharmaceutical composition is formulated as a tablet or a
capsule.
[0100] The crystalline forms of the present disclosure can be
delivered directly or in pharmaceutical compositions along with
suitable excipients, as is well known in the art. Various
treatments embodied herein can comprise administration of an
effective amount of a crystalline form of the disclosure to a
subject in need, e.g., a subject having or at risk for anemia due
to, e.g., chronic renal failure, diabetes, cancer, AIDS, radiation
therapy, chemotherapy, kidney dialysis, or surgery. In one
embodiment, the subject is a mammalian subject, and in one
embodiment, the subject is a human subject.
[0101] An effective amount of a crystalline form can readily be
determined by routine experimentation, as can the most effective
and convenient route of administration and the most appropriate
formulation. In one embodiment, the dosage may be from 0.05 mg/kg
to about 700 mg/kg per day. Typically, the dosage may be from about
0.1 mg/kg to about 500 mg/kg; from about 0.5 mg/kg to about 250
mg/kg; from about 1 mg/kg to about 100 mg/kg; from about 1 mg/kg to
about 10 mg/kg; from about 1 mg/kg to about 5 mg/kg; or from about
1 mg/kg to about 2 mg/kg. For example, the dosage may be about 1.0
mg/kg; about 1.2 mg/kg; about 1.5 mg/kg; about 2.0 mg/kg; or about
2.5 mg/kg. Various formulations and drug delivery systems are
available in the art (see, e.g., Gennaro, A. R., ed. (1995)
Remington's Pharmaceutical Sciences).
[0102] Suitable routes of administration may, for example, include
oral, rectal, transmucosal, nasal, or intestinal administration and
parenteral delivery, including intramuscular, subcutaneous,
intramedullary injections, as well as intrathecal, direct
intraventricular, intravenous, intraperitoneal, intranasal, or
intraocular injections. The crystalline form or composition thereof
may be administered in a local rather than a systemic manner. For
example, a crystalline form or composition thereof can be delivered
via injection or in a targeted drug delivery system, such as a
depot or sustained release formulation. In one embodiment, the
route of administration is oral.
[0103] The pharmaceutical compositions of the present disclosure
may be manufactured by any of the methods well-known in the art,
such as by conventional mixing, dissolving, granulating,
dragee-making, levigating, emulsifying, encapsulating, entrapping,
or lyophilizing processes. As noted above, the compositions can
include one or more pharmaceutically acceptable excipients that
facilitate processing of active molecules into preparations for
pharmaceutical use.
[0104] Proper formulation is dependent upon the route of
administration chosen. For injection, for example, the composition
may be formulated in aqueous solutions, preferably in
physiologically compatible buffers such as Hanks' solution,
Ringer's solution, or physiological saline buffer. For transmucosal
or nasal administration, penetrants appropriate to the barrier to
be permeated are used in the formulation. Such penetrants are
generally known in the art. In a preferred embodiment of the
present disclosure, the present crystalline forms are prepared in a
formulation intended for oral administration. For oral
administration, it can be formulated readily by combining the
crystalline forms with pharmaceutically acceptable excipients well
known in the art. Such excipients enable the crystalline forms of
the disclosure to be formulated as tablets, pills, dragees,
capsules, liquids, gels, syrups, slurries, suspensions and the
like, for oral ingestion by a subject. The crystalline forms may
also be formulated in rectal compositions such as suppositories or
retention enemas, e.g., containing conventional suppository bases
such as cocoa butter or other glycerides.
[0105] Pharmaceutical preparations for oral use can be obtained
using solid excipients, optionally grinding a resulting mixture,
and processing the mixture of granules, after adding suitable
auxiliaries, if desired, to obtain tablets or dragee cores.
Suitable excipients are, for example, fillers such as sugars,
including lactose, sucrose, mannitol, or sorbitol; cellulose
preparations such as, for example, maize starch, wheat starch, rice
starch, potato starch, gelatin, gum tragacanth, methyl cellulose,
hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose,
microcrystalline cellulose and/or polyvinylpyrrolidone (PVP or
povidone). If desired, disintegrating agents may be added, such as
the cross-linked polyvinyl pyrrolidone, agar, croscarmellose sodium
or alginic acid or a salt thereof such as sodium alginate. Also,
wetting agents such as sodium dodecyl sulfate or lubricants such as
magnesium stearate may be included.
[0106] Dragee cores are provided with suitable coatings. For this
purpose, concentrated sugar solutions may be used, which may
optionally contain gum arabic, talc, polyvinyl pyrrolidone,
carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer
solutions, and suitable organic solvents or solvent mixtures.
Dyestuffs or pigments may be added to the tablets or dragee
coatings for identification or to characterize different
combinations of active doses.
[0107] Pharmaceutical preparations for oral administration include
push-fit capsules made of gelatin, as well as soft, sealed capsules
made of gelatin and a plasticizer, such as glycerol or sorbitol.
The push-fit capsules can contain the active ingredients in
admixture with filler such as lactose, binders such as starches,
and/or lubricants such as talc or magnesium stearate and,
optionally, stabilizers. In soft capsules, the crystalline forms
may be dissolved or suspended in suitable liquids, such as fatty
oils, liquid paraffin, or liquid polyethylene glycols. In addition,
stabilizers may be added. All formulations for oral administration
should be in dosages suitable for such administration.
[0108] In one embodiment, the crystalline forms described herein
can be administered transdermally, such as through a skin patch, or
topically. In one aspect, the transdermal or topical formulations
can additionally comprise one or multiple penetration enhancers or
other effectors, including agents that enhance migration of the
delivered compound. Transdermal or topical administration could be
preferred, for example, in situations in which location specific
delivery is desired.
[0109] For administration by inhalation, the crystalline forms for
use according to the present disclosure are conveniently delivered
in the form of an aerosol spray presentation from pressurized packs
or a nebulizer, with the use of a suitable propellant, e.g.,
dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane, carbon dioxide, or any other suitable
gas. In the case of a pressurized aerosol, the appropriate dosage
unit may be determined by providing a valve to deliver a metered
amount. Capsules and cartridges of, for example, gelatin, for use
in an inhaler or insufflator may be formulated. These typically
contain a powder mix of the crystalline form and a suitable powder
base such as lactose or starch.
[0110] Compositions formulated for parenteral administration by
injection, e.g., by bolus injection or continuous infusion can be
presented in unit dosage form, e.g., in ampoules or in multi-dose
containers, with an added preservative. The compositions may take
such forms as suspensions, solutions, or emulsions in oily or
aqueous vehicles, and may contain formulatory agents such as
suspending, stabilizing and/or dispersing agents. Formulations for
parenteral administration include aqueous solutions or other
compositions in water-soluble form.
[0111] Suspensions of the crystalline forms may also be prepared as
appropriate oily injection suspensions. Suitable lipophilic
solvents or vehicles include fatty oils such as sesame oil and
synthetic fatty acid esters, such as ethyl oleate or triglycerides,
or liposomes. Aqueous injection suspensions may contain substances
that increase the viscosity of the suspension, such as sodium
carboxymethyl cellulose, sorbitol, or dextran. Optionally, the
suspension may also contain suitable stabilizers or agents that
increase the solubility of the crystalline forms to allow for the
preparation of highly concentrated solutions. Alternatively, the
active ingredient may be in powder form for constitution with a
suitable vehicle, e.g., sterile pyrogen-free water, before use.
[0112] As mentioned above, the compositions of the present
disclosure may also be formulated as a depot preparation. Such long
acting formulations may be administered by implantation (for
example, subcutaneously or intramuscularly) or by intramuscular
injection. Thus, for example, the present crystalline forms may be
formulated with suitable polymeric or hydrophobic materials (for
example as an emulsion in an acceptable oil) or ion exchange
resins, or as sparingly soluble derivatives, for example, as a
sparingly soluble salt.
[0113] For any composition used in the various treatments embodied
herein, a therapeutically effective dose can be estimated initially
using a variety of techniques well known in the art. For example,
in a cell culture assay, a dose can be formulated in animal models
to achieve a circulating concentration range that includes the
IC.sub.50 as determined in cell culture. Dosage ranges appropriate
for human subjects can be determined, for example, using data
obtained from cell culture assays and non-human animal studies. In
one embodiment, the dosage may be from 0.05 mg/kg to about 700
mg/kg administered periodically. The dosage may be administered
once a day, every other day, one, two or three times a week, or at
other appropriate intervals as can be readily determined by
competent medical practitioners. Typically the dosage is
administered 2 or 3 times a week. Typically, the dosage may be from
about 0.1 mg/kg to about 500 mg/kg; from about 0.5 mg/kg to about
250 mg/kg; from about 1 mg/kg to about 100 mg/kg; from about 1
mg/kg to about 10 mg/kg; from about 1 mg/kg to about 5 mg/kg; or
from about 1 mg/kg to about 2 mg/kg. For example, the dosage may be
about 1.0 mg/kg; about 1.2 mg/kg; about 1.5 mg/kg; about 2.0 mg/kg;
or about 2.5 mg/kg.
[0114] A therapeutically effective dose of a compound refers to
that amount of the compound that results in amelioration of
symptoms or a prolongation of survival in a subject. Toxicity and
therapeutic efficacy of such molecules can be determined by
standard pharmaceutical procedures in cell cultures or experimental
animals, e.g., by determining the LD.sub.50 (the dose lethal to 50%
of the population) and the ED.sub.50 (the dose therapeutically
effective in 50% of the population). The dose ratio of toxic to
therapeutic effects is the therapeutic index, which can be
expressed as the ratio LD.sub.50/ED.sub.50. Compounds that exhibit
high therapeutic indices are preferred.
[0115] Dosages preferably fall within a range of circulating
concentrations that includes the ED.sub.50 with little or no
toxicity. Dosages may vary within this range depending upon the
dosage form employed and the route of administration utilized. The
exact formulation, route of administration, and dosage should be
chosen, according to methods known in the art, in view of the
specifics of a subject's condition.
[0116] Dosage amount and interval may be adjusted individually to
provide plasma levels of the active moiety that are sufficient to
modulate a desired parameter, e.g., endogenous erythropoietin
plasma levels, i.e. minimal effective concentration (MEC). The MEC
will vary for each compound but can be estimated from, for example,
in vitro data. Dosages necessary to achieve the MEC will depend on
individual characteristics and route of administration. Compounds
or compositions thereof should be administered using a regimen
which maintains plasma levels above the MEC for about 10-90% of the
duration of treatment, preferably about 30-90% of the duration of
treatment, and most preferably between 50-90%. In cases of local
administration or selective uptake, the effective local
concentration of the drug may not be related to plasma
concentration. Alternatively, modulation of a desired parameter,
e.g., stimulation of endogenous erythropoietin, may be achieved by
1) administering a loading dose followed by a maintenance dose, 2)
administering an induction dose to rapidly achieve the desired
parameter, e.g., erythropoietin levels, within a target range,
followed by a lower maintenance dose to maintain, e.g., hematocrit,
within a desired target range, or 3) repeated intermittent
dosing.
[0117] The amount of compound or composition administered will, of
course, be dependent on a variety of factors, including the sex,
age, and weight of the subject being treated, the severity of the
affliction, the manner of administration, and the judgment of the
prescribing physician.
[0118] The present compositions may, if desired, be presented in a
pack or dispenser device containing one or more unit dosage forms
containing the active ingredient. Such a pack or device may, for
example, comprise metal or plastic foil, such as a blister pack.
The pack or dispenser device may be accompanied by instructions for
administration. Compositions comprising a crystalline form of the
disclosure formulated in a compatible pharmaceutical excipient may
also be prepared, placed in an appropriate container, and labeled
for treatment of an indicated condition. Suitable conditions
indicated on the label may include treatment of conditions,
disorders, or diseases in which anemia is a major indication.
4. Method of Use
[0119] One aspect of the disclosure provides for use of one or more
of a crystalline or amorphous form of
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid (Compound A), or a composition comprising one or more
crystalline or amorphous forms of Compound A or a solvate or salt
thereof, for the manufacture of a medicament for use in treating
various conditions or disorders as described herein. It also
provides methods of using the crystalline or amorphous form, or
composition or medicament thereof, to treat, pretreat, or delay
progression or onset of various conditions or disorders as
described herein. In one embodiment, the crystalline form of
Compound A used in the method is Form A. In one embodiment, the
crystalline form of Compound A used in the method is Form B, Form C
or Form D. As used in this section "Method of Use" and the
corresponding method claims, "compound" refers to a crystalline or
amorphous form of Compound A, or a solvate or salt thereof.
[0120] In one embodiment, at least about 85% of the compound used
in the method is Compound A Form A. In one embodiment, at least
about 90% of the compound used in the method is Compound A Form A.
In one embodiment, at least about 95% of the compound used in the
method is Compound A Form A. In one embodiment, at least about 99%
of the compound used in the method is Compound A Form A. In one
embodiment, at least about 99.5% of the compound used in the method
is Compound A Form A. In one embodiment, at least about 99.9% of
the compound used in the method is Compound A Form A. In one
embodiment, at least about 99.99% of the compound used in the
method is Compound A Form A.
[0121] The medicaments or compositions can be used to modulate the
stability and/or activity of HIF, and thereby activate
HIF-regulated gene expression. The crystalline or amorphous form,
or composition or medicament thereof, can be used in methods to
treat, pretreat, or delay progression or onset of conditions
associated with HIF including, but not limited to, anemic,
ischemic, and hypoxic conditions. In various embodiments, the
crystalline or amorphous form, or composition or medicament
thereof, is administered immediately following a condition
producing acute ischemia, e.g., myocardial infarction, pulmonary
embolism, intestinal infarction, ischemic stroke, and renal
ischemic-reperfusion injury. In another embodiment, the crystalline
or amorphous form, or composition or medicament thereof, is
administered to a patient diagnosed with a condition associated
with the development of chronic ischemia, e.g., cardiac cirrhosis,
macular degeneration, pulmonary embolism, acute respiratory
failure, neonatal respiratory distress syndrome, and congestive
heart failure. In yet another embodiment, the crystalline or
amorphous form, or composition or medicament thereof, is
administered immediately after a trauma or injury. In other
embodiments, the crystalline or amorphous form, or composition or
medicament thereof, can be administered to a subject based on
predisposing conditions, e.g., hypertension, diabetes, occlusive
arterial disease, chronic venous insufficiency, Raynaud's disease,
chronic skin ulcers, cirrhosis, congestive heart failure, and
systemic sclerosis. In still other embodiments, the crystalline or
amorphous form, or composition or medicament thereof, may be
administered to pretreat a subject to decrease or prevent the
development of tissue damage associated with ischemia or
hypoxia.
[0122] The crystalline or amorphous form, or compositions or
medicaments thereof, can also be used to increase endogenous
erythropoietin (EPO). The crystalline or amorphous form, or
composition or medicament thereof, can be administered to prevent,
pretreat, or treat EPO-associated conditions, including, e.g.,
conditions associated with anemia and neurological disorders.
Conditions associated with anemia include disorders such as acute
or chronic kidney disease, diabetes, cancer, ulcers, infection with
virus, e.g., HIV, bacteria, or parasites; inflammation, etc. Anemic
conditions can further include those associated with procedures or
treatments including, e.g., radiation therapy, chemotherapy,
dialysis, and surgery. Disorders associated with anemia
additionally include abnormal hemoglobin and/or erythrocytes, such
as found in disorders such as microcytic anemia, hypochromic
anemia, aplastic anemia, etc.
[0123] The disclosure is also directed to use of a crystalline or
amorphous form, or composition or medicament thereof, to treat,
pretreat, or delay onset of a condition associated with a disorder
selected from the group consisting of anemic disorders;
neurological disorders and/or injuries including cases of stroke,
trauma, epilepsy, and neurodegenerative disease; cardiac ischemia
including, but not limited to, myocardial infarction and congestive
heart failure; liver ischemia including, but not limited to,
cardiac cirrhosis; renal ischemia including, but not limited to,
acute kidney failure and chronic kidney failure; peripheral
vascular disorders, ulcers, burns, and chronic wounds; pulmonary
embolism; and ischemic-reperfusion injury.
[0124] The disclosure is also directed to a method of inhibiting
the activity of at least one hydroxylase enzyme which modifies the
alpha subunit of hypoxia inducible factor. The HIF hydroxylase
enzyme may be a prolyl hydroxylase including, but not limited to,
the group consisting of EGLN1, EGLN2, and EGLN3 (also known as
PHD2, PHD1 and PHD3, respectively), described by Taylor (2001, Gene
275:125-132), and characterized by Aravind and Koonin (2001, Genome
Biol 2:RESEARCH0007), Epstein et al. (2001, Cell 107:43-54), and
Bruick and McKnight (2001, Science 294:1337-1340). The method
comprises contacting the enzyme with an inhibiting effective amount
of one or more crystalline or amorphous form of Compound A. In some
embodiments, the HIF hydroxylase enzyme is an asparaginyl
hydroxylase or a prolyl hydroxylase. In other embodiments, the HIF
hydroxylase enzyme is a factor inhibiting HIF, human EGLN1, EGLN2,
or EGLN3.
[0125] While this disclosure has been described in conjunction with
specific embodiments and examples, it will be apparent to a person
of ordinary skill in the art, having regard to that skill and this
disclosure, that equivalents of the specifically disclosed
materials and methods will also be applicable to this disclosure;
and such equivalents are intended to be included within the
following claims.
Examples
[0126] Unless otherwise stated, the following abbreviations used
throughout the specification have the following definitions:
TABLE-US-00001 .degree. C. Degree Celsius Ac Acetyl ca. About d
Doublet dd Doublet of doublets DMA Dimethylamine DMEM Eagle's
minimal essential medium DMF Dimethylformamide DMSO
Dimethylsulfoxide DSC Differential scanning calorimetry EDTA
Ethylenediaminetetraacetic acid EtOAc Ethyl Acetate eq. Equivalents
FBS Fetal bovine serum g Gram Gly Glycine h Hour HEPES
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid HPLC High
performance liquid chromatography IPA Isopropyl alcohol iPrOAc
Isopropylacetate J Joules J Coupling constant kg Kilogram kV
Kilovolts m Multiplet M Molar M+ Mass peak mA Milliamps Me Methyl
MEC Minimal effective concentration MeCN Acetonitrile MEK Methyl
ethyl ketone mg Milligram MHz Megahertz MIBK Methyl iso-butyl
ketone min Minute mIU Milliintemational units mL Milliliter mm
Millimeter mM Millimolar mol Mole MS Mass spectroscopy NMR Nuclear
magnetic resonance PBS Phosphate buffer system Ph Phenyl RH
Relative humidity rpm Revolutions per minute s Singlet s Second TEA
Triethylamine TGA Thermogravimetric analysis THF Tetrahydrofuran Ts
Tosyl vol Volume w weight XRPD X-ray powder diffraction .delta.
Chemical shift .mu.L Microliter .mu.M Micromolar
X-Ray Powder Diffraction (XRPD)
[0127] X-Ray Powder Diffraction patterns were collected on a Bruker
AXS C2 GADDS diffractometer using Cu K.alpha. radiation (40 kV, 40
mA), automated XYZ stage, laser video microscope for auto-sample
positioning and a HiStar 2-dimensional area detector. X-ray optics
consists of a single Gobel multilayer mirror coupled with a pinhole
collimator of 0.3 mm. A weekly performance check is carried out
using a certified standard NIST 1976 Corundum (flat plate).
[0128] The beam divergence, i.e. the effective size of the X-ray
beam on the sample, was approximately 4 mm. A .theta.-.theta.
continuous scan mode was employed with a sample-detector distance
of 20 cm which gives an effective 2.theta. range of
3.2.degree.-29.7.degree.. Typically the sample would be exposed to
the X-ray beam for 120 seconds. The software used for data
collection was GADDS for WNT 4.1.16 and the data were analyzed and
presented using Diffrac Plus EVA v11.0.0.2 or v13.0.0.2.
[0129] Alternatively, X-Ray Powder Diffraction patterns were
collected on a Bruker D8 diffractometer using Cu K.alpha. radiation
(40 kV, 40 mA), .theta.-2.theta. goniometer, and divergence of V4
and receiving slits, a Ge monochromator and a Lynxeye detector. The
instrument is performance checked using a certified Corundum
standard (NIST 1976). The software used for data collection was
Diffrac Plus XRD Commander v2.5.0 and the data were analyzed and
presented using Diffrac Plus EVA v11.0.0.2 or v13.0.0.2.
[0130] Samples were run under ambient conditions as flat plate
specimens using powder as received. The sample was gently packed
into a cavity cut into polished, zero-background (510) silicon
wafer. The sample was rotated in its own plane during analysis. The
details of the data collection are: [0131] Angular range: 2 to
42.degree.2.theta. [0132] Step size: 0.05.degree.2.theta. [0133]
Collection time: 0.5 s/step [0134] Analysis duration: 7 minutes
Differential Scanning Calorimetry (DSC)
[0135] DSC was were collected on a TA Instruments Q2000 equipped
with a 50 position autosampler. The calibration for thermal
capacity was carried out using sapphire and the calibration for
energy and temperature was carried out using certified indium.
Typically 0.5-3 mg of each sample, in a pin-holed aluminum pan, was
heated at 10.degree. C./min from 25.degree. C. to 300.degree. C. A
purge of dry nitrogen at 50 ml/min was maintained over the sample.
Modulated temperature DSC was carried out using an underlying
heating rate of 2.degree. C./min and temperature modulation
parameters of .+-.0.318.degree. C. (amplitude) every 60 seconds
(period). The instrument control software was Advantage for Q
Series v2.8.0.392 and Thermal Advantage v4.8.3 and the data were
analyzed using Universal Analysis v4.4A.
[0136] Alternatively, the DSC data was collected on a Mettler DSC
823e equipped with a 34 position auto-sampler. The instrument was
calibrated for energy and temperature using certified indium.
Typically 0.5-3 mg of each sample, in a pin-holed aluminum pan, was
heated at 10.degree. C./min from 25.degree. C. to 300.degree. C. or
25.degree. C. to 320.degree. C. A nitrogen purge at 50 ml/min was
maintained over the sample. The instrument control and data
analysis software was STARe v9.20.
Thermo-Gravimetric Analysis (TGA)
[0137] TGA data were collected on a Mettler TGNSDT A 851 e equipped
with a 34 position autosampler. The instrument was temperature
calibrated using certified indium. Typically, 1-30 mg of each
sample was loaded onto a pre-weighed aluminum crucible and was
heated at 10.degree. C./min from ambient temperature to 350.degree.
C. A nitrogen purge at 50 ml/min was maintained over the sample.
The instrument control and data analysis software was STARe
v9.20.
Example 1. Preparation of Compound A Form A
Methods
[0138] The crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid (Compound A Form A) was prepared via the following
methods.
Method I
[0139] The crystalline Compound A Form A (see Example 1, Method I)
was used in this method. 15 mg of the crystalline material was used
with each solvent added in increments until a clear solution had
been obtained or until 50 volumes (750 .mu.L) of solvent had been
added. Samples were sonicated for 5 seconds after each solvent
addition. When insoluble, the slurries were stirred at 500 rpm
cycling between 25.degree. C. and 50.degree. C. (4 h at each
temperature) for a period of from 16 hours to six days. Any
resulting solutions were then allowed to evaporate at room
temperature. The solids obtained from this experiment were analyzed
by XRPD.
[0140] Each of the following solvents used in the above described
Method I provided Form A: acetic acid, acetone, acetophenone,
benzonitrile, benzyl alcohol, butyronitrile, chlorobenzene,
cyclohexanone, 1,2-dichlorobenzene, 1,2-dichloroethane,
dimethoxyethane, dimethylacetamide, DMSO, 1,4-dioxane, ethylene
glycol, EtOAc, formamide, hexafluorobenzene, hexane, IPA, IPA:10%
water, iPrOAc, MeCN, MEK, MIBK, nitromethane, perfluorohexane,
propionitrile, sulfolane, t-butyl methyl ether, t-butanol,
tetraline, THF, and toluene.
[0141] Using Method I, hexafluoropropan-2-ol, methanol and ethanol
did not provide Form A.
Method II
[0142] The crystalline Compound A Form A (see Example 1, Method
VIII) was used in this method. 15 mg of the crystalline material
was used with 30 volumes (450 .mu.L) of solvent with the exception
of DMSO and DMA where 5 volumes were used. Slurries were sonicated
for 5 seconds. The slurries were stirred at 500 rpm at 5.degree. C.
for a period of six days. Any resulting solutions were then allowed
to evaporate at room temperature. The solids obtained were analyzed
by XRPD.
[0143] Each of the following solvents used in the above described
Method II provided Form A: benzonitrile, sulfolane, formamide,
tetraline, acetophenone, benzyl alcohol, ethylene glycol,
1,2-dichlorobenzene, chlorobenzene, cyclohexanone, butyronitrile,
acetic acid, nitromethane, propionitrile, dimethoxyethane,
1,2-dichloroethane, hexafluorobenzene, t-butanol, hexane, and
perfluorohexane.
[0144] Using Method II, hexafluoropropan-2-ol did not provide Form
A.
Method III
[0145] The crystalline Compound A Form A (see Example 1, Method
VIII) was used in this method. Method III is substantially as
described in Method II, above, with the exception that the slurries
were stirred at 500 rpm at 50.degree. C. for a period of six days.
The solids obtained were analyzed by XRPD.
[0146] Each of the following solvents used in the above described
Method III provided Form A: benzonitrile, sulfolane, formamide,
tetraline, acetophenone, benzyl alcohol, ethylene glycol,
1,2-dichlorobenzene, chlorobenzene, cyclohexanone, butyronitrile,
acetic acid, nitromethane, propionitrile, dimethoxyethane,
1,2-dichloroethane, hexafluorobenzene, t-butanol, hexane, and
perfluorohexane.
[0147] Using Method III, dimethylacetamide, t-butyl methyl ether,
and hexafluoropropan-2-ol did not provide Form A.
Method IV
[0148] The crystalline Compound A Form B (see Example 2) was used
in this method. 15 mg of the crystalline material was used with 30
volumes (450 .mu.L) solvent with the exception of DMSO and DMA
where 5 volumes was used. Slurries were sonicated for 5 seconds.
The slurries were stirred at 500 rpm, cycling between 25.degree. C.
and 50.degree. C. (4 h at each temperature) for six days. Any
resulting solutions were then left to evaporate quickly at room
temperature. The solids were analyzed by XRPD.
[0149] Each of the following solvents used in the above described
Method IV provided Form A: benzonitrile, sulfolane, formamide,
tetraline, acetophenone, benzyl alcohol, ethylene glycol,
1,2-dichlorobenzene, chlorobenzene, cyclohexanone, butyronitrile,
acetic acid, t-butyl methyl ether, nitromethane, propionitrile,
dimethoxyethane, 1,2-dichloroethane, hexafluorobenzene, t-butanol,
hexane, perfluorohexane, and hexafluoropropan-2-ol.
Method V
[0150] The crystalline Compound A Form A (see Example 1, Method
VIII) was used in this method. 20 mg of the crystalline material
was dissolved in THF (410 .mu.L) before the addition of 10 volumes
(200 .mu.L) of solvent with the exception of DMSO and DMA where 5
volumes was used. The slurries were stirred at 500 rpm cycling
between 25.degree. C. and 50.degree. C. (4 h at each temperature)
for 48 hours. Any solutions which were obtained after the heat/cool
cycles were allowed to evaporate at room temperature. The solids
obtained were analyzed by XRPD.
[0151] Each of the following solvents used in the above described
Method V provided Form A. Benzonitrile, sulfolane, formamide,
tetraline, acetophenone, benzyl alcohol, ethylene glycol, DMSO,
1,2-dichlorobenzene, chlorobenzene, cyclohexanone, butyronitrile,
acetic acid, t-butyl methyl ether, propionitrile, dimethoxyethane,
1,2-dichloroethane, hexafluorobenzene, t-butanol, and hexane.
[0152] Using Method V, nitromethane, hexafluoropropan-2-ol, and
perfluorohexane did not provide Form A.
Method VI
[0153] The crystalline Compound A Form A (30 mg, see Example 1,
Method VIII) was dissolved in 10 mL of acetone. This solution was
subject to fast solvent evaporation on a rota-evaporator
(40.degree. C., 35-50 Torr). 12.85 mg of the resulting material was
used with 10 volumes (128.5 .mu.L) of solvent with the exception of
DMSO and DMA where 5 volumes was used. Slurries were sonicated for
5 seconds. The slurries were stirred at 500 rpm between 25.degree.
C. and 50.degree. C. (8 h cycles) for a period of six days. Any
resulting solutions were then allowed to evaporate at room
temperature. Solids obtained were analyzed by XRPD.
[0154] Each of the following solvents used in the above described
Method VI provided Form A: benzonitrile, sulfolane, formamide,
tetraline, acetophenone, benzyl alcohol, ethylene glycol, DMSO,
1,2-dichlorobenzene, chlorobenzene, butyronitrile, acetic acid,
t-butyl methyl ether, nitromethane, propionitrile, dimethoxyethane,
1,2-dichloroethane, hexafluorobenzene, t-butanol, and hexane.
[0155] Using Method VI, cyclohexanone, hexafluoropropan-2-ol, and
perfluorohexane did not provide Form A.
Method VII
[0156] The crystalline Compound A Form A (see Example 1, Method
VIII) was used in this method. 30 mg was suspended in 7 volumes of
solvent (10% aqueous). Slurries were sonicated for 5 seconds. The
slurries were stirred at 500 rpm cycling between 25.degree. C. and
50.degree. C. (8 h cycles) for a period of four days. The solids
obtained were analyzed by XRPD.
[0157] Each of the following solvents used in the above described
Method VII provided Form A: acetone, acetonitrile, ethanol,
methanol, 2-methyl-THF, and IPA.
Method VIII
[0158] An aqueous solution of sodium hydroxide was added slowly to
a stirred suspension of Compound A in water at temperature range
(10.degree. C. to 90.degree. C.). A solution of acetic acid in
water was then slowly charged at temperature range (10.degree. C.
to 90.degree. C.) and the mixture was stirred. The solid was
filtered, washed with water, and dried under vacuum to constant
weight. Compound A Form A was obtained as white to light yellow
crystalline solid.
Data
[0159] The XRPD pattern for Compound A Form A is shown in FIG. 1
and peaks and their related intensities in the XRPD pattern are
shown in Table 1 below.
TABLE-US-00002 TABLE 1 Peaks in the XRPD Pattern for Compound A
Form A Peak Position (.degree.2.theta.) Relative Intensity (%) 8.5
100 10.1 3.5 11.4 9.2 12.8 20.6 14.5 3.2 15.9 13.4 16.2 45.5 16.9
18.5 17.1 11.5 17.5 19.0 19.0 12.5 19.9 7.7 20.2 2.8 21.6 31.9 21.8
16.0 22.0 11.9 22.2 17.2 22.6 17.4 22.9 36.4 23.6 4.7 23.8 6.5 24.1
3.4 24.7 11.0 25.2 3.0 25.6 9.8 25.8 16.5 27.4 60.6 28.2 7.7 28.4
3.7 29.1 7.6 29.2 5.8 29.6 5.3 30.0 2.7 30.4 2.3 31.3 2.8 31.9 5.9
32.0 6.1 32.8 3.0 33.4 15.6 33.6 16.1 34.1 5.2 34.6 2.8 35.1 4.3
35.2 4.2 35.3 3.2 35.7 4.0 36.5 2.2 36.6 2.2 36.9 2.4 37.0 2.4 37.3
4.0 37.4 2.8 37.7 2.3 37.8 2.3 38.2 2.9 38.5 3.2 38.9 2.4 39.3 2.4
40.8 2.8 41.5 4.9
[0160] The results of the differential scanning calorimetry and
thermogravimetric analyses of Compound A Form A are presented in
FIGS. 2 and 24, respectively. The thermogravimetric analysis shows
negligible weight loss of approximately 0.4% between 25.degree. C.
and 225.degree. C., followed by a steady loss of weight above
225.degree. C., suggesting sublimation or decomposition of the
material at these temperatures (FIG. 24). The differential scanning
calorimetry analysis of Compound A Form A showed a very shallow
exotherm in the range from about 80-190.degree. C., followed by a
sharp endotherm at about 224.3.degree. C. (peak maximum). The sharp
endotherm corresponded to the melt of the material, as determined
by hotstage microscopy.
[0161] The hotstage microscopy of Compound A Form A showed little
change of the material below its melting point. Some changes in
birefringence were noted in the range of about 150-200.degree. C.
The sample melted within the temperature range of about
218.5-222.4.degree. C.
[0162] The moisture sorption data for Compound A Form A showed
negligible weight gain, approximately 0.2% gained from between 5%
to 95% relative humidity, which was lost on desorption. The small
moisture uptake of Compound A Form A is indicative of a kinetically
non-hygroscopic material.
Example 2. Preparation of Compound A Form B
Method
[0163] Crystalline Compound A Form B was provided by lyophilization
of Form A in a 1,4-dioxane:water (2:1) mixture. 20 mg of the
crystalline Compound A Form B was dissolved in 20 volumes
1,4-dioxane before addition of 20 volumes cosolvent. The solvent
systems were left to evaporate at room temperature, under the fume
hood. The solids obtained from this experiment were analysed by
XRPD.
[0164] Each of the following cosolvents used in the above described
method provided Form B. 1,4-Dioxane:water (1:1), 1,4-dioxane:water
(1:1), 1,4-dioxane:methanol (1:1), 1,4-dioxane:ethanol,
1,4-dioxane:acetone (1:1), 1,4-dioxane:THF (1:1), and
1,4-dioxane:heptane (1:1).
Data
[0165] The XRPD pattern for Compound A Form B is shown in FIG. 3
and peaks and their related intensities in the XRPD pattern are
shown in Table 2 below. The crystalline pattern changed after the
sample was stored at 25.degree. C./96% RH for twelve days,
reverting to Form A.
TABLE-US-00003 TABLE 2 Peaks in the XRPD Pattern for Compound A
Form B Peak Position (.degree.2.theta.) Relative Intensity (%) 4.2
53.9 8.3 100 9.8 1.3 10.5 1.1 11.5 1.0 12.5 12.9 12.7 8.3 12.9 1.3
14.1 13.7 15.8 3.9 16.6 76.3 17.4 10.4 19.2 3.7 20.9 8.2 21.0 3.7
21.8 3.7 22.7 3.0 22.9 4.6 24.9 2.7 25.0 4.9 25.9 1.5 27.5 1.5 28.4
2.1 28.8 1.7 29.3 9.0 30.0 1.3 30.8 1.4 31.6 1.2 33.5 6.9 33.6 9.9
35.2 1.5 37.0 1.2 37.9 4.3 41.6 0.8
[0166] No residual solvent was observed, other than water. A weight
loss of 2.8% w/w between room temperature and 90.degree. C. in the
TGA thermogram suggested the presence of 0.5 equivalent of water
(theoretical 2.5% w/w) (FIG. 4). The DSC thermogram showed an
endothermic event associated to the weight loss (FIG. 4). A sharp
endothermic event occurs at 222.3.degree. C. (-127.8 J/g), which
matches the melt of Form A.
[0167] High resolution XRPD data was collected over a month. After
a full month at ambient temperature (32 days), the sample (Form B)
almost completely reverted to anhydrous Form A.
Example 3. Preparation of Compound A Form C
Method
[0168] Crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid hexafluoropropan-2-ol solvate (Compound A Form C) was prepared
following the procedure described in Methods I, II, III and VI of
Example 1 using hexafluoropropan-2-ol as the solvent.
Data
[0169] The XRPD pattern for Compound A Form C is shown in FIG. 5
and peaks and their related intensities in the XRPD pattern are
shown in Table 3 below.
TABLE-US-00004 TABLE 3 Peaks in the XRPD Pattern for Compound A
Form C Peak Position (.degree.2.theta.) Relative Intensity (%) 4.5
100 8.5 1.6 9.1 7.9 10.2 1.9 11.2 2.5 11.8 1.2 12.5 1.2 13.7 37
15.4 9.2 15.5 8.3 15.6 3 16.4 11.5 16.9 2.5 17.7 3.3 18.3 2.7 18.7
1.2 19.0 2.3 19.8 2.5 20.6 8 21.9 3.5 22.4 1.5 22.5 2.3 22.9 6.3
23.3 3.1 23.8 3.4 24.6 2 25.2 2.3 25.4 3.1 25.7 1.9 26.0 1.5 27.5
1.2 27.9 2.3 28.4 2.6 29.1 1.4 29.4 1 29.8 1 30.2 1 30.8 2.1 31.6
1.2 31.7 1.5 32.2 1.3 32.3 1.1 33.1 0.9 34.1 1.4 34.2 1.5 35.1 0.9
35.5 0.8 37.0 0.9 37.8 1.1 38.7 1 40.2 0.7 40.9 1.1 41.8 1.2
[0170] Residual solvent was observed by proton NMR and was assigned
to the hexafluoropropan-2-ol. Thermal analysis was also carried out
on this sample (FIG. 6). A weight loss of 7.8% w/w between room
temperature and 130.degree. C. in the TGA thermogram suggested the
presence of 1/6 equivalent of hexafluoropropan-2-ol (theoretical
7.36% w/w) in the sample. The DSC thermogram showed an endothermic
event associated to the weight loss followed by a small exothermic
event (ca. 130.degree. C.) (FIG. 6). A sharp endothermic event
occurs at 222.2.degree. C. (-17.9 J/g), which matches the melt of
Form A. In conclusion, the material isolated from
hexafluoropropan-2-ol is a meta-stable solvate under ambient
conditions and converts to Form A.
Example 4. Preparation of Compound A Form D
Method
[0171] Crystalline
[(4-hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid DMSO:water solvate (Compound A Form D) was prepared from slow
evaporation from THF/DMSO (20 volumes THF/5 volumes DMSO) of either
mostly amorphous Compound A or Form A.
Data
[0172] The XRPD pattern for Compound A Form D is shown in FIG. 8
and peaks and their related intensities in the XRPD pattern are
shown in Table 4 below. The sample (Form D) converted to Form A
when left drying at ambient temperature.
TABLE-US-00005 TABLE 4 Peaks in the XRPD Pattern for Compound A
Form D Peak Position (.degree.2.theta.) Relative Intensity (%) 4.2
32.7 4.3 29.4 8.4 65 8.5 65 12.6 30.9 15.8 20.6 16.8 100 18.9 5.5
19.3 19.1 20.7 6.2 21.7 3.8 22.1 23 22.6 4.6 23.1 11.7 24.0 8.7
24.5 3.1 25.3 12.1 25.4 10.3 26.8 4 27.1 4.4 27.3 3.9 28.4 43 31.2
7.7 32.0 3.4 33.3 4.4 33.4 3 34.0 6.2 35.8 3.7 38.3 7.6 39.1 3.6
40.9 2.5 41.6 3.6
[0173] Residual solvent was observed by proton NMR and was assigned
to the DMSO. The TGA thermogram and DSC thermogram are shown in
FIG. 8. The TGA shows a first weight loss between 40-150.degree. C.
of 18.5% (combination of water and DMSO) and a second weight loss
between 170-220.degree. C. (possible DMSO). Two broad endotherms at
37.6.degree. C. and 90.4.degree. C. were possible due to loss of
water and DMSO. A small endotherm at 222.0.degree. C. was observed
due to melt of Form A.
Example 5. Preparation of Salts of Compound A
Method
[0174] The crystalline Compound A Form A was used to prepare the
following salts. Compound A Form A (50 mg per experiment) was
dissolved in acetone or THF (50 vol, 2.1 ml) at 50.degree. C. The
solutions were treated with 1.1 mol eq. of the corresponding
counter ion (for example, 1.0 M aqueous solution of sodium
hydroxide, potassium hydroxide or hydrochloric acid). The
temperature was maintained at 50.degree. C. for 20 min then cooled
to 0.degree. C. at 0.1.degree. C./min with stirring. After 20 h at
0.degree. C., the solids were filtered, air dried for 10 min and
analyzed by the appropriate techniques.
Data
Compound A Sodium Salt
[0175] The XRPD pattern for Compound A sodium salt is shown in FIG.
9 and peaks and their related intensities in the XRPD pattern are
shown in Table 5 below.
TABLE-US-00006 TABLE 5 Peaks in the XRPD Pattern for Compound A
Sodium Salt Peak Position (.degree.2.theta.) Relative Intensity (%)
5.3 22.0 8.1 4.4 11.1 10.9 13.5 9.3 15.1 5.2 16.0 100.0 17.0 9.6
18.7 20.0 19.2 13.3 21.6 30.7 22.9 7.7 24.0 12.6 25.3 7.5 26.2 11.0
28.9 10.1
[0176] The stoichiometry (ionic Compound A:counter ion) was
determined to be 1:1 by Ion Chromatography (Metrohm 761 Compact IC,
IC Net software v2.3). The TGA thermogram and DSC thermogram are
shown in FIG. 10. The TGA thermogram shows a weight loss between
40-90.degree. C. of 11.5%. The DSC thermogram shows a broad
endotherm at 64.1.degree. C., followed by two exotherms at
150.5.degree. C. and 190.3.degree. C. and a sharp melt at
313.6.degree. C. Purity was determined to be about 99.6%.
Compound A Potassium Salt
[0177] The stoichiometry (ionic Compound A:counter ion) was
determined to be 1:1 by Ion Chromatography (Metrohm 761 Compact IC,
IC Net software v2.3). The XRPD pattern for the Compound A
potassium salt is shown in FIG. 21. As seen in the Figure, the
potassium salt is substantially amorphous. Thermal analysis showed
a possible loss of water followed by a recrystallization event to
produce non-solvated, crystalline form with a melt at 291.degree.
C. (FIG. 22).
Compound a L-Arginine Salt
[0178] The XRPD pattern for Compound A L-arginine salt is shown in
FIG. 11 and peaks and their related intensities in the XRPD pattern
are shown in Table 6 below.
TABLE-US-00007 TABLE 6 Peaks in the XRPD Pattern for Compound A
L-arginine salt Peak Position (.degree.20) Relative Intensity (%)
10.8 24.5 11.4 13.1 12.0 15.1 15.8 31.7 17.1 18.2 18.0 20.9 19.4
41.9 20.8 100.0 21.8 59.7 22.7 53.7 23.4 56.8 24.4 37.5 25.4 64.4
26.4 53.7 26.8 48.8 27.5 52.3 28.5 40.4 29.7 30.5
[0179] The stoichiometry (ionic Compound A:counter ion) was
determined to be about 1:1 by NMR. The TGA thermogram and DSC
thermogram are shown in FIG. 12. The TGA thermogram shows a weight
loss between 40-100.degree. C. of 4.5%. The DSC thermogram shows
two broad endotherms at 79.6.degree. C. and 143.3.degree. C., an
exotherm at 172.5.degree. C., followed by an endotherm at
210.1.degree. C. Purity was determined to be about 99.5%.
Compound a L-Lysine Salt
[0180] The XRPD pattern for Compound A L-lysine salt is shown in
FIG. 13 and peaks and their related intensities in the XRPD pattern
are shown in Table 7 below.
TABLE-US-00008 TABLE 7 Peaks in the XRPD Pattern for Compound A
L-lysine salt Peak Position (.degree.2.theta.) Relative Intensity
(%) 6.8 11.5 9.5 36.4 9.9 57.4 10.2 71.5 13.1 13.9 13.8 18.4 14.4
44.8 14.6 41.6 16.9 78.8 17.3 55.4 18.4 65.1 19.0 50.1 19.8 92.4
20.7 100.0 21.2 95.1 22.1 53.7 23.6 58.0 25.5 59.8 25.0 64.7 26.1
63.9 27.4 54.3 28.4 53.4 28.6 53.8 28.8 52.3 30.0 41.7 30.6
38.2
[0181] The stoichiometry (ionic Compound A:counter ion) was
determined to be about 1:1 by NMR. The TGA thermogram and DSC
thermogram are shown in FIG. 14. The TGA thermogram shows a weight
loss between 235-270.degree. C. of 12.1%. The DSC thermogram shows
a sharp melt at 230.7.degree. C. and a broad endotherm at
237.1.degree. C. Purity was determined to be about 99.6%.
Compound a Ethanolamine Salt
[0182] The XRPD patterns for Compound A ethanolamine salt is shown
in FIG. 15 and peaks and their related intensities in the XRPD
patterns are shown in Tables 8, 9 and 10 below. Pattern 1 was
observed from acetone, pattern 2 was observed from THF, and Pattern
3 was observed at 40.degree. C./75% RH.
TABLE-US-00009 TABLE 8 Peaks in the XRPD Pattern for Compound A
ethanolamine salt (Pattern 1) Peak Position (.degree.2.theta.)
Relative Intensity (%) 3.8 15.6 4.6 6.3 5.1 8.3 7.7 4.8 10.9 32.8
12.5 10.0 15.0 28.2 15.5 23.5 17.9 14.0 18.6 20.1 21.1 45.2 21.8
54.9 22.7 77.7 24.4 30.3 26.2 53.6 26.6 47.6 27.1 100.0
TABLE-US-00010 TABLE 9 Peaks in the XRPD Pattern for Compound A
ethanolamine salt (Pattern 2) Peak Position (.degree.2.theta.)
Relative Intensity (%) 12.5 25.5 13.1 22.4 14.9 12.8 15.9 37.4 16.7
29.3 17.1 68 0 17.8 19.4 18.5 79.6 20.2 42.4 21.4 73.3 22.8 100.0
23.8 80.2 24.7 34.6 25.7 57.9 26.8 28.9 27.4 19.0 28.0 32.7 29.5
41.4 30.8 19.6
TABLE-US-00011 TABLE 10 Peaks in the XRPD Pattern for Compound A
ethanolamine salt (Pattern 3) Peak Position (.degree.2.theta.)
Relative Intensity (%) 7.2 18.1 10.3 15.2 10.8 39.7 13.4 8.1 14.1
37.3 16.2 52.9 16.9 37.5 18.1 17.7 21.3 100.0 21.7 64.8 22.3 21.7
22.9 42.3 23.1 39.6 23.7 36.1 25.3 29.9 26.2 23.3 26.9 50.5 27.8
75.0 29.0 22.7 18.5 11.8
[0183] For both THF and acetone, the stoichiometry (ionic Compound
A:counter ion) was determined to be about 1:1 by NMR. The TGA
thermogram and DSC thermogram for the Compound A ethanolamine salt
from acetone is shown in FIG. 16. The TGA thermogram shows a weight
loss between 155-250.degree. C. of 10.1% (0.8 equivalents of
ethanolamine). The DSC thermogram shows a sharp melt at
171.4.degree. C. and a broad endotherm at 186.0.degree. C. Purity
was determined to be about 99.0%. For the Compound A ethanolamine
salt from THF, the TGA thermogram showed a weight loss between
155-250.degree. C. of 10.1% (0.8 equivalents of ethanolamine), and
the DSC thermogram showed a sharp melt at 172.4.degree. C. and a
broad endotherm at 185.5.degree. C. Purity was determined to be
about 99.1%.
Compound a Diethanolamine Salt
[0184] The XRPD patterns for Compound A diethanolamine salt is
shown in FIG. 17 and peaks and their related intensities in the
XRPD patterns are shown in Tables 11 and 12 below. Pattern 1 was
observed from acetone and Pattern 2 was observed at 40.degree.
C./75% RH.
TABLE-US-00012 TABLE 11 Peaks in the XRPD Pattern for Compound A
diethanolamine salt (Pattern 1) Peak Position (.degree.2.theta.)
Relative Intensity (%) 6.6 6.6 11.2 12.5 11.8 21.6 13.0 9.5 14.5
13.6 15.6 22.9 16.9 100.0 19.6 37.5 20.5 27.7 21.4 23.1 22.6 37.3
23.7 42.9 25.0 46.9 26.0 36.5 27.1 35.3 28.3 20.8 29.4 17.1 30.6
13.2
TABLE-US-00013 TABLE 12 Peaks in the XRPD Pattern for Compound A
diethanolamine salt (Pattern 2) Peak Position (.degree.2.theta.)
Relative Intensity (%) 5.9 9.4 8.9 5.9 11.1 23.9 11.5 13.9 11.9
12.7 13.9 7.0 14.9 27.7 16.0 100.0 18.3 20.0 20.0 11.9 20.9 32.5
21.5 20.5 22.3 26.4 23.3 54.6 24.0 17.1 24.8 33.5 25.6 22.4 27.6
27.7 29.0 20.1
[0185] The stoichiometry (ionic Compound A:counter ion) was
determined to be about 1:1 by NMR. The TGA thermogram and DSC
thermogram are shown in FIG. 18. The TGA thermogram shows a weight
loss between 155-250.degree. C. of 12.6%. The DSC thermogram shows
a sharp melt at 150.2.degree. C. and a broad endotherm at
172.2.degree. C. Purity was determined to be about 99.7%.
Compound a Tromethamine Salt
[0186] The XRPD pattern for Compound A tromethamine salt is shown
in FIG. 19 and peaks and their related intensities in the XRPD
pattern is shown in Table 13 below.
TABLE-US-00014 TABLE 13 Peaks in the XRPD Pattern for Compound A
tromethamine salt Peak Position (.degree.2.theta.) Relative
Intensity (%) 9.4 4.3 10.1 100.0 11.8 8.5 13.4 15.3 14.2 53.0 15.0
5.1 16.9 20.3 19.2 12.9 20.1 23.5 21.1 69.2 22.3 21.4 23.4 14.7
24.3 13.6 25.1 10.5 25.7 21.5 26.3 16.9 28.4 22.9 29.2 12.0 30.0
17.1
[0187] The stoichiometry (ionic Compound A:counter ion) was
determined to be about 1:1 by NMR. The TGA thermogram and DSC
thermogram are shown in FIG. 20. The TGA thermogram shows a weight
loss between 180-260.degree. C. of 11.0%. The DSC thermogram shows
a sharp melt at 176.5.degree. C. and a broad endotherm at
182.6.degree. C. Purity was determined to be about 99.7%.
Compound a Hydrochloric Acid Salt
[0188] The XRPD pattern for Compound A hydrochloric acid salt is
shown in FIG. 25 and peaks and their related intensities in the
XRPD pattern is shown in Table 14 below.
TABLE-US-00015 TABLE 14 Peaks in the XRPD Pattern for Compound A
hydrochloric acid salt Peak Position (.degree.2.theta.) Relative
Intensity (%) 8.2 6.4 10.2 18.3 10.7 13.6 12.5 10.9 13.6 36.1 19.0
27.1 19.8 21.1 20.3 30.6 20.9 42.7 21.3 17.4 22.5 68.5 24.1 100.0
27.6 36.4 29.0 17.9 25.6 21.5
[0189] The stoichiometry (ionic Compound A:counter ion) was
determined to be about 1:1 by Ion Chromatography. The TGA
thermogram and DSC thermogram are shown in FIG. 26. The TGA
thermogram shows a weight loss between 100-170.degree. C. of 6.5%
and a second weight loss between 185-210.degree. C. of 3.4%. The
DSC thermogram shows two small endotherms at 154.3 and
201.6.degree. C., and a sharp melt at 223.0.degree. C. Purity was
determined to be about 99.1%.
Compound a Sulfuric Acid Salt
[0190] The XRPD pattern for Compound A sulfuric acid salt is shown
in FIG. 27 to be a mixture of the salt and Form A. The peaks and
their related intensities in the XRPD pattern is shown in Table 15
below.
TABLE-US-00016 TABLE 15 Peaks in the XRPD Pattern for Compound A
sulfuric acid salt Peak Position (.degree.2.theta.) Relative
Intensity (%) 6.1 48.8 8.5 36.2 15.4 15.0 16.1 82.4 17.1 32.4 17.4
75.9 19.8 82.9 22.9 100.0 23.7 33.4 24.7 99.5 26.1 73.0 27.3 70.9
28.1 29.8 28.8 35.1 29.6 35.1
[0191] The TGA thermogram and DSC thermogram are shown in FIG. 28.
The TGA thermogram shows a weight loss between 10-110.degree. C. of
6.5% and a second weight loss between 180-280.degree. C. of 27.4%.
The DSC thermogram shows three small endotherms at 31.8, 55.7 and
91.0 associated with the first weight loss, and a large, broad
endotherm at 201.4.degree. C. due to decomposition.
Compound a Methanesulfonic Acid Salt
[0192] The XRPD patterns for Compound A methanesulfonic acid salt
is shown in FIG. 29 and the peaks and their related intensities in
the XRPD patterns are shown in Tables 16 and 17 below. Pattern 1
was observed from acetone, and pattern 2 was observed from THF.
Pattern 1 reverted to Form A at 40.degree. C./75% RH and pattern 2
reverted to a mixture of Form A and Pattern 1 at 40.degree. C./75%
RH.
TABLE-US-00017 TABLE 16 Peaks in the XRPD Pattern for Compound A
methanesulfonic acid salt (Pattern 1) Peak Position
(.degree.2.theta.) Relative Intensity (%) 4.9 100.0 9.8 21.0 12.9
24.9 14.9 25.8 17.0 43.9 19.4 49.7 21.5 25.3 23.7 23.4 24.8
21.5
TABLE-US-00018 TABLE 17 Peaks in the XRPD Pattern for Compound A
methanesulfonic acid salt (Pattern 2) Peak Position
(.degree.2.theta.) Relative Intensity (%) 4.6 25.8 6.9 18.4 7.1
12.7 9.7 8.3 12.2 26.6 11.6 8.8 13.6 15.5 14.0 20.1 17.7 31.7 18.7
100.0 20.5 93.8 22.1 44.8 23.1 24.7 23.7 30.7 24.6 54.3 25.6 30.3
26.4 21.6 27.0 36.7 28.4 17.2 29.3 25.4 30.5 14.1
[0193] For both THF and acetone, the stoichiometry (ionic Compound
A:counter ion) was determined to be about 1:1 by NMR. The TGA
thermogram and DSC thermogram for the Compound A methanesulfonic
acid salt from THF is shown in FIG. 30. The TGA thermogram shows a
weight loss between 40-100.degree. C. of 2.1%. The DSC thermogram
shows a small endotherm at 153.5.degree. C. and a sharp endotherm
at 166.9.degree. C. Purity was determined to be about 99.3%. For
the Compound A methanesulfonic acid salt from acetone, the TGA
thermogram showed no weight loss until sample degradation at about
180.degree. C., and the DSC thermogram showed an endotherm at
144.5.degree. C. due to sample melt. Purity was determined to be
about 98.9%.
Example 6. Preparation of Amorphous Compound a
[0194] Amorphous Compound A was obtained as follows. Crystalline
Compound A Form A (500 mg) was dissolved in THF (1.5 mL) at room
temperature. The solution was filtered to remove any residual
crystalline material. The solvent was removed by fast evaporation
in the rotary evaporator. An aliquot of the obtained solid was
examined by XRPD. Alternatively, amorphous Compound A was obtained
by lyophilization of Compound A Form A from a mixture of
1,4-dioxane:water (2:1 v/v) or by packing and sealing Compound A
into a capillary tube, melting the sample on a Kofler hotbench at
about 240.degree. C. for one minute and cooling at ambient
temperature. An XRPD of amorphous Compound A is shown in FIG.
23.
Example 7. Preparation of Bis TEA Salt of Compound a
Method
[0195] The crystalline Compound A Form A (50 mg) was dissolved in
acetone (50 vol) at 50.degree. C. The solution was treated with 2.1
mol eq. of triethylamine (TEA). The temperature was maintained at
50.degree. C. for 20 min then cooled to 0.degree. C. at 0.1.degree.
C./min with stirring. After 72 at 0.degree. C., the solids were
filtered, air dried for 5 min and analysed by XRPD. The solutions
were set for slow evaporation at ambient conditions.
Data
[0196] The XRPD pattern for Compound A bis TEA salt is shown in
FIG. 31 and peaks and their related intensities in the XRPD pattern
are shown in Table 18 below.
TABLE-US-00019 TABLE 18 Peaks in the XRPD Pattern for Compound A
bis triethylamine salt Peak Position (.degree.2.theta.) Relative
Intensity (%) 5.5 4.2 9.2 5.0 10.1 29.4 11.7 13.2 13.3 8.0 13.7
17.3 14.5 42.5 14.7 52.1 15.2 9.3 15.7 14.3 16.3 16.0 17.0 36.9
18.4 11.9 19.4 31.5 20.4 21.7 20.7 27.3 22.6 100.0 22.9 23.9 23.9
29.9 24.8 14.6 25.5 38.5 26.3 16.7 27.2 37.1 28.1 62.2
[0197] The TGA thermogram and DSC thermogram are shown in FIG. 32.
The TEA was isolated as a bis salt, however the salt started to
dissociate to Form A after one week storage at 40.degree. C./75%
RH.
Example 8. Preparation of Hemi Calcium and Magnesium Salts
Method
[0198] The calcium and magnesium salts of Compound A were prepared
by ion exchange from the sodium salt. Compound A (450 mg) was
dissolved in acetone at 50.degree. C. (50 vol, 22.5 ml). The
solution was treated with 1.1 mol eq. of sodium hydroxide (1 M
solution in water). A suspension was formed on addition then it was
cooled to 0.degree. C. at 0.1.degree. C./min. After 48 h at
0.degree. C., the solid was filtered, air dried for 10 min and
analysed by XRPD. The sodium salt (50 mg) was dissolved in MeOH (20
vol, 1 ml) at room temperature. The solution was heated to
50.degree. C. and then treated with the corresponding counter ion
(1 M solutions in MeOH). The mixtures were cooled to 0.degree. C.
at 0.1.degree. C./min. After 24 h at 0.degree. C., the solids were
filtered, air dried for 5 min and analyzed by XRPD (first crop).
The liquors were kept and set for slow evaporation at ambient
conditions to provide a second crop. The materials isolated from
the Ca.sup.+2 and Mg.sup.+2 hemi salt experiments crystallized
after one week incubation at 40.degree. C./75% RH.
Data
[0199] The XRPD patterns for the Compound A hemi calcium and
magnesium salts are shown in FIG. 33 and FIG. 35, respectively. The
peaks and their related intensities in the XRPD patterns are shown
in Tables 19 and 20 below.
TABLE-US-00020 TABLE 19 Peaks in the XRPD Pattern for Compound A
hemi calcium salt Peak Position (.degree.2.theta.) Relative
Intensity (%) 5.7 8.8 6.6 10.5 8.6 10.2 9.7 8.1 10.9 24.0 11.5 63.5
12.9 36 4 13.4 20.5 14.4 32.8 14.6 34.0 15.3 20.8 17.0 27.6 17.7
19.6 20.0 29.0 21.6 31.0 23.1 51.9 23.8 39.2 24.4 52.4 25.9 100.0
27.0 88.1 28.1 40.3 29.3 25.7
TABLE-US-00021 TABLE 20 Peaks in the XRPD Pattern for Compound A
hemi magnesium salt Peak Position (.degree.2.theta.) Relative
Intensity (%) 5.0 12.7 7.1 37.2 7.5 24.9 8.0 30.0 10.0 30.2 11.4
35.4 11.9 37.5 12.6 33.9 '12.9 37.5 15.9 45.4 17.8 50.4 18.9 44.4
20.1 67.6 21.0 59.0 21.7 67.9 22.8 100.0
[0200] The DSC thermograms for the Compound A hemi calcium and
magnesium salts are shown in FIG. 34 and FIG. 36, respectively.
Example 9. Single Crystal of Compound A Form A
[0201] Single crystals were grown from acetone with sufficient
quality for structure determination by single crystal X-ray
diffraction.
[0202] The structure solution was obtained by direct methods,
full-matrix least-squares refinement on F.sup.2 with weighting
w.sup.-1=.sigma.2(F.sub.o.sup.2)+(0.0697 P).sup.2+(0.3149 P), where
P=(F.sub.o.sup.2+2F.sub.c.sup.2)/3, anisotropic displacement
parameters, empirical absorption correction using spherical
harmonics, implemented in SCALE3 ABSPACK scaling algorithm. Final
wR.sup.2={.SIGMA.[w(F.sub.o.sup.2-2F.sub.c.sup.2).sup.2]/.SIGMA.[w(F.sub.-
o.sup.2).sup.2].sup.1/2}=0.1376 for all data, conventional
R.sub.1=0.0467 on F values of 2496 reflections with
F.sub.o>4.sigma.(F.sub.o), S=1.045 for all data and 248
parameters. Final .DELTA./.sigma.(max) 0.000,
.DELTA./.sigma.(mean), 0.000. Final difference map between +0.211
and -0.318 e .ANG..sup.-3.
[0203] FIG. 37 shows a view of a molecule of Compound A Form A from
the crystal structure showing the numbering scheme employed.
Anisotropic atomic displacement ellipsoids for the non-hydrogen
atoms are shown at the 50% probability level. Hydrogen atoms are
displayed with an arbitrarily small radius.
TABLE-US-00022 TABLE 21 Samples Submitted for Single Crystal X-Ray
Diffraction Studies Molecular Formula
C.sub.19H.sub.16N.sub.2O.sub.5 Molecular Weight 352.34 Crystal
system Triclinic Space group P-1 a 8.5208(13) .ANG., .alpha.
98.415(11).degree., b 9.2233(13) .ANG., .beta. 108.788(12).degree.,
c 11.1859(14) .ANG. .gamma. 102.841(12).degree. V 788.50(19)
.ANG..sup.3 Z 2 D.sub.c 1.484 g cm.sup.-1 .mu. 0.909 mm.sup.-1
Source, .lamda. Cu-K.alpha., 1.54178 .ANG. F(000) 368 T 100(2) K
crystal Colorless prism, 0.11 .times. 0.05 .times. 0.02 mm data
truncated to 0.80 .ANG. .theta..sub.max 77.18.degree. Completeness
98.2% Reflections 12441 Unique reflections 3282 R.sub.int
0.0406
Example 10. Preparation of Compound A
a) 5-Phenoxyphthalide
##STR00003##
[0205] A reactor was charged with DMF (68 Kg), and stirring was
initiated. The reactor was then charged with phenol (51 Kg),
acetylacetone (8 Kg), 5-bromophthalide (85 Kg), copper bromide (9
Kg), and potassium carbonate (77 Kg). The mixture was heated above
85.degree. C. and maintained until reaction completion and then
cooled. Water was added. Solid was filtered and washed with water.
Solid was dissolved in dichloromethane, and washed with aqueous HCl
and then with water. Solvent was removed under pressure and
methanol was added. The mixture was stirred and filtered. Solid was
washed with methanol and dried in an oven giving 5-phenoxyphthalide
(Yield: 72%, HPLC: 99.6%).
b) 2-Chloromethyl-4-phenoxybenzoic acid methyl ester
##STR00004##
[0207] A reactor was charged with toluene (24 Kg), and stirring was
initiated. The reactor was then charged with 5-phenoxyphthalide (56
Kg), thionyl chloride (41 Kg), trimethyl borate (1 Kg),
dichlorotriphenylphosphorane (2.5 Kg), and potassium carbonate (77
Kg). The mixture was heated to reflux until reaction completion and
solvent was removed leaving 2-chloromethyl-4-phenoxybenzoyl
chloride. Methanol was charged and the mixture was heated above
50.degree. C. until reaction completion. Solvent was removed and
replaced with DMF. This solution of the product methyl
2-chloromethyl-4-phenoxybenzoic acid methyl ester in DMF was used
directly in the next step (HPLC: 85%).
c) 4-Hydroxy-7-phenoxyisoquinoline-3-carboxylic acid methyl ester
(1a)
##STR00005##
[0209] A reactor was charged with a solution of
2-chloromethyl-4-phenoxybenzoic acid methyl ester (.about.68 Kg) in
DMF, and stirring was initiated. The reactor was then charged with
p-toluenesulfonylglycine methyl ester (66 Kg), potassium carbonate
(60 Kg), and sodium iodide (4 Kg). The mixture was heated to at
least 50.degree. C. until reaction completion. The mixture was
cooled. Sodium methoxide in methanol was charged and the mixture
was stirred until reaction completion. Acetic acid and water were
added, and the mixture was stirred, filtered and washed with water.
Solid was purified by acetone trituration and dried in an oven
giving 1a (Yield from step b): 58%; HPLC: 99.4%). .sup.1H NMR (200
MHz, DMSO-d6) .delta. 11.60 (s, 1H), 8.74 (s, 1H), 8.32 (d, J=9.0
Hz, 1H), 7.60 (dd, J=2.3 & 9.0 Hz, 1H), 7.49 (m, 3H), 7.24 (m,
3H), 3.96 (s, 3H); MS-(+)-ion M+1=296.09
d) Methyl
1-((dimethylamino)methyl)-4-hydroxy-7-phenoxyisoquinoline-3-carb-
oxylate (1b)
##STR00006##
[0211] A flask was charged with 1a (29.5 g) and acetic acid (44.3
g.+-.5%), and then stirred. Bis-dimethylaminomethane (12.8 g.+-.2%)
was slowly added. The mixture was heated to 55.+-.5.degree. C. and
maintained until reaction completion. The reaction product was
evaluated by MS, HPLC and .sup.1H NMR. .sup.1H NMR (200 MHz,
DMSO-d6) .delta. 11.7 (s, 1H), 8.38 (d, J=9.0 Hz, 1H), 7.61 (dd,
J=9.0, 2.7 Hz, 1H), 7.49 (m, 3H), 7.21 (m, 3H), 5.34 (s, 2H), 3.97
(s, 3H), 1.98 (s, 3H); MS-(+)-ion M+1=368.12.
e) Methyl
1-((acetoxy)methyl)-4-hydroxy-7-phenoxyisoquinoline-3-carboxylat- e
(1c)
##STR00007##
[0213] The solution of 1b from a) above was cooled below 25.degree.
C., at which time acetic anhydride (28.6 g.+-.3.5%) was added to
maintain temperature below 50.degree. C. The resulting mixture was
heated to 100.+-.5.degree. C. until reaction completion.
[0214] The solution of 1c and 1d from above was cooled to less than
65.+-.5.degree. C. Water (250 mL) was slowly added. The mixture was
then cooled to below 20.+-.5.degree. C. and filtered. The wet cake
was washed with water (3.times.50 mL) and added to a new flask.
Dichloromethane (90 mL) and water (30 mL) were added, and the
resulting mixture was stirred. The dichloromethane layer was
separated and evaluated by HPLC.
[0215] The organic layer was added to a flask and cooled
5.+-.5.degree. C. Morpholine was added and the mixture was stirred
until reaction completion. Solvent was replaced with
acetone/methanol mixture. After cooling, compound 1c precipitated
and was filtered, washed and dried in an oven (Yield: 81%, HPLC:
>99.7%). .sup.1H NMR (200 MHz, DMSO-d6) .delta. 11.6 (S, 1H),
8.31 (d, J=9.0 Hz, 1H), 7.87 (d, J=2.3 Hz, 1H), 7.49 (m, 3H), 7.24
(m, 3H), 3.95 (s, 3H), 3.68 (s, 2H), 2.08 (s, 6H); MS-(+)-ion
M+1=357.17.
f) Methyl 4-hydroxy-1-methyl-7-phenoxyisoquinoline-3-carboxylate
(1e)
##STR00008##
[0217] A reactor was charged with 1c (16.0 g), Pd/C (2.08 g),
anhydrous Na.sub.2CO.sub.3 (2.56 g) and ethyl acetate (120 mL). The
flask was vacuum-purged with nitrogen (3.times.) and vacuum-purged
with hydrogen (3.times.). The flask was then pressurized with
hydrogen and stirred at about 60.degree. C. until completion of
reaction. The flask was cooled to 20-25.degree. C., the pressure
released to ambient, the head space purged with nitrogen three
times and mixture was filtered. The filtrate was concentrated.
Methanol was added. The mixture was stirred and then cooled.
Product precipitated and was filtered and dried in an oven (Yield:
90%, HPLC: 99.7%).
g)
[(4-Hydroxy-1-methyl-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic
acid (Compound A)
##STR00009##
[0219] A pressure flask was charged with 1e (30.92 g), glycine
(22.52 g), methanol (155 mL), sodium methoxide solution (64.81 g)
and sealed (as an alternative, sodium glycinate was used in place
of glycine and sodium methoxide). The reaction was heated to about
110.degree. C. until reaction was complete. The mixture was cooled,
filtered, washed with methanol, dried under vacuum, dissolved in
water and washed with ethyl acetate. The ethyl acetate was removed
and to the resulting aqueous layer an acetic acid (18.0 g) solution
was added. The suspension was stirred at room temperature,
filtered, and the solid washed with water (3.times.30 mL), cold
acetone (5-10.degree. C., 2.times.20 mL), and dried under vacuum to
obtain Compound A (Yield: 86.1%, HPLC: 99.8%).
Example 11. Biological Testing
[0220] The solid forms provided herein can be used for inhibiting
HIF hydroxylase activity, thereby increasing the stability and/or
activity of hypoxia inducible factor (HIF), and can be used to
treat and prevent HIF-associated conditions and disorders (see,
e.g., U.S. Pat. No. 7,323,475, U.S. Patent Application Publication
No. 2007/0004627, U.S. Patent Application Publication No.
2006/0276477, and U.S. Patent Application Publication No.
2007/0259960, incorporated by reference herein).
[0221] The biological activity of the solid forms provided herein
may be assessed using any conventionally known method. In
particular embodiments, cells derived from animal tissues,
preferably human tissues, capable of expressing erythropoietin when
stimulated by compounds of the invention are cultured for the in
vitro production of endogenous proteins. Cells contemplated for use
in such methods include, but are not limited to, cells derived from
hepatic, hematopoietic, renal, and neural tissues.
[0222] Cell culture techniques are generally available in the art
and include any method that maintains cell viability and
facilitates expression of endogenous proteins. Cells are typically
cultured in a growth medium optimized for cell growth, viability,
and protein production. Cells may be in suspension or attached to a
substrate, and medium may be supplied in batch feed or continuous
flow-through regimens. Compounds of the invention are added to the
culture medium at levels that stimulate erythropoietin production
without compromising cell viability. Erythropoietin produced by the
cells is secreted into the culture medium. The medium is then
collected and the erythropoietin is purified using methods known to
those of skill in the art. (See, e.g., Lai et al. (1987) U.S. Pat.
No. 4,667,016; and Egrie (1985) U.S. Pat. No. 4,558,006.)
[0223] Suitable assay methods are well known in the art. The
following are presented only as examples and are not intended to be
limiting.
Cell-Based HIF.alpha. Stabilization Assay
[0224] Human cells (e.g., Hep3B cells from hepatocellular tissue)
derived from various tissues were separately seeded into 35 mm
culture dishes, and grown at 37.degree. C., 20% O.sub.2, 5%
CO.sub.2 in standard culture medium, e.g., DMEM (Dulbecco's
modification of Eagle's medium), 10% FBS (fetal bovine serum). When
cell layers reach confluence, the media was replaced with OPTI-MEM
media (Invitrogen Life Technologies, Carlsbad Calif.), and cell
layers were incubated for approximately 24 hours in 20% O.sub.2, 5%
CO.sub.2 at 37.degree. C. Compound A or 0.013% DMSO (dimethyl
sulfoxide) was then added to existing medium and incubation was
continued overnight.
[0225] Following incubation, the media was removed, centrifuged,
and stored for analysis (see Cell-based VEGF and EPO assays below).
The cells were washed two times in cold phosphate buffered saline
(PBS) and then lysed in 1 mL of 10 mM Tris (pH 7.4), 1 mM EDTA, 150
mM NaCl, 0.5% IGEPAL (Sigma-Aldrich, St. Louis Mo.), and a protease
inhibitor mix (Roche Molecular Biochemicals) for 15 minutes on ice.
Cell lysates were centrifuged at 3,000.times.g for 5 minutes at
4.degree. C., and the cytosolic fractions (supernatant) were
collected. The nuclei (pellet) were resuspended and lysed in 100
.mu.L of 20 mM HEPES (pH 7.2), 400 mM NaCl, 1 mM EDTA, 1 mM
dithiothreitol, and a protease mix (Roche Molecular Biochemicals),
centrifuged at 13,000.times.g for 5 minutes at 4.degree. C., and
the nuclear protein fractions (supernatant) were collected.
[0226] The Nuclear protein fractions collected were analyzed for
HIF-1a using a QUANTIKINE immunoassay (R&D Systems, Inc.,
Minneapolis Minn.) according to the manufacturer's
instructions.
Cell-Based EPO Assay
[0227] Hep3B cells (human hepatocellular carcinoma cells from ATCC,
cat # HB-8064) were plated at 25,000 cells per well 96 well plates.
The next day, the cells were washed once with DMEM (Cellgro, cat
#10-013-CM)+0.5% fetal bovine serum (Cellgro, cat #35-010-CV) and
incubated with various concentrations of compound or vehicle
control (0.15% DMSO) in DMEM+0.5% fetal bovine serum for 72 hours.
Cell free culture supernatants were generated by transfer to a
conical bottom 96 well plate and centrifugation for 5 minutes at
2000 rpm. The supernatant was quantitated for EPO using a human EPO
ELISA kit (R&D Systems, cat # DEP 00).
[0228] The EPO values for the compounds reported herein (e.g.,
Table 22) are the measured value for cells plus compound minus the
value for the vehicle control for the same cell preparation. The
EPO values for the vehicle control for the cell preparations varied
from 0-12.5 mIU/mL.
HIF-PH Assay
[0229] Ketoglutaric acid .alpha.-[1-.sup.14C]-sodium salt,
alpha-ketoglutaric acid sodium salt, and HPLC purified peptide were
obtained from commercial sources, e.g., Perkin-Elmer (Wellesley
MA), Sigma-Aldrich, and SynPep Corp. (Dublin Calif.), respectively.
Peptides for use in the assay were fragments of HIF.alpha. as
described above or as disclosed in International Publication WO
2005/118836, incorporated by reference herein. For example, a HIF
peptide for use in the HIF-PH assay was
[methoxycoumarin]-DLDLEALAPYIPADDDFQL-amide. HIF-PH, e.g., HIF-PH2
(also known as EGLN1 or PHD2), was expressed in, e.g., insect Hi5
cells, and partially purified, e.g., through a SP ion exchange
chromatography column. Enzyme activity was determined by capturing
.sup.14CO.sub.2 using an assay described by Kivirikko and Myllyla
(1982, Methods Enzymol. 82:245-304). Assay reactions contained 50
mM HEPES (pH 7.4), 100 .mu.M .alpha.-ketoglutaric acid sodium salt,
0.30 .mu.Ci/mL .alpha.-ketoglutaric acid
.alpha.-[1-.sup.14C]-sodium salt, 40 .mu.M FeSO.sub.4, 1 mM
ascorbate, 1541.8 units/mL Catalase, with or without 50 .mu.M
peptide substrate and various concentrations of compound of the
invention. Reactions were initiated by addition of HIF-PH
enzyme.
[0230] The peptide-dependent percent turnover was calculated by
subtracting percent turnover in the absence of peptide from percent
turnover in the presence of substrate peptide. Percent inhibition
and IC.sub.50 were calculated using peptide-dependent percent
turnover at given inhibitor concentrations. Calculation of
IC.sub.50 values for each inhibitor was conducted using GraFit
software (Erithacus Software Ltd., Surrey UK). The results are
summarized in Table 22.
[0231] Table 21 below was intended to demonstrate the
pharmacological utility of Compound A. By inhibiting HIF prolyl
hydroxylase enzymes (for example PHD2, also known as EGLN1),
Compound A stabilizes HIF.alpha., which then combines with
HIF.beta. to form an active transcription factor that increases
expression of numerous genes involved in response to hypoxic and
ischemic conditions, including erythropoietin (EPO). Therefore
Compound A can be used for the prevention, pretreatment, or
treatment of conditions associated with HIF and or EPO including
anemic, ischemic and hypoxic conditions.
TABLE-US-00023 TABLE 22 IC.sub.50 Cell EPO* PHD2 (.mu.M) (mIU/mL)
Compound A Form A 2.1 182 *Cell EPO measured at 30 .mu.M compound
in DMSO compared to DMSO only control
* * * * *