U.S. patent application number 13/646130 was filed with the patent office on 2013-04-18 for pharmaceutical compositions of 3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl) cyclopropanecarboxamido)-3-methylpyriodin-2-yl)benzoic acid and administration thereof.
This patent application is currently assigned to VERTEX PHARMACEUTICALS INCORPORATED. The applicant listed for this patent is VERTEX PHARMACEUTICALS INCORPORATED. Invention is credited to Rossitza Gueorguieva Alargova, Irina Nikolaevna Kadiyala, Ritu Rohit Kaushik, Marinus Jacobus Verwijs, Christopher Ryan Young.
Application Number | 20130095181 13/646130 |
Document ID | / |
Family ID | 47075277 |
Filed Date | 2013-04-18 |
United States Patent
Application |
20130095181 |
Kind Code |
A1 |
Verwijs; Marinus Jacobus ;
et al. |
April 18, 2013 |
PHARMACEUTICAL COMPOSITIONS OF
3-(6-(1-(2,2-DIFLUOROBENZO[D][1,3]DIOXOL-5-YL)
CYCLOPROPANECARBOXAMIDO)-3-METHYLPYRIODIN-2-YL)BENZOIC ACID AND
ADMINISTRATION THEREOF
Abstract
A pharmaceutical composition comprising Compound 1,
(3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3-
-methylpyridin-2-yl)benzoic acid), and at least one excipient
selected from: a filler, a diluent, a disintegrant, a surfactant, a
binder, a glidant and a lubricant, the composition being suitable
for oral administration to a patient in need thereof to treat a
CFTR mediated disease such as Cystic Fibrosis. Methods for treating
a patient in need thereof include administering an oral
pharmaceutical formulation of Compound 1 to the patient.
Inventors: |
Verwijs; Marinus Jacobus;
(Framingham, MA) ; Alargova; Rossitza Gueorguieva;
(Brighton, MA) ; Kaushik; Ritu Rohit; (Watertown,
MA) ; Kadiyala; Irina Nikolaevna; (Newton, MA)
; Young; Christopher Ryan; (Waltham, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
VERTEX PHARMACEUTICALS INCORPORATED; |
CAMBRIDGE |
MA |
US |
|
|
Assignee: |
VERTEX PHARMACEUTICALS
INCORPORATED
CAMBRIDGE
MA
|
Family ID: |
47075277 |
Appl. No.: |
13/646130 |
Filed: |
October 5, 2012 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
PCT/US2011/031519 |
Apr 7, 2011 |
|
|
|
13646130 |
|
|
|
|
61366562 |
Jul 22, 2010 |
|
|
|
61321729 |
Apr 7, 2010 |
|
|
|
61321748 |
Apr 7, 2010 |
|
|
|
Current U.S.
Class: |
424/465 ;
424/400; 514/312; 514/338 |
Current CPC
Class: |
A61K 9/141 20130101;
A61K 9/2077 20130101; A61K 9/2866 20130101; C07D 451/02 20130101;
A61P 19/10 20180101; A61P 7/00 20180101; A61K 31/47 20130101; A61K
9/28 20130101; A61P 21/00 20180101; A61P 21/02 20180101; A61P 35/00
20180101; A61P 15/10 20180101; A61P 25/00 20180101; A61P 3/10
20180101; A61P 25/02 20180101; A61P 7/04 20180101; A61K 9/1623
20130101; A61P 43/00 20180101; C07D 405/12 20130101; A61P 13/02
20180101; A61P 25/08 20180101; A61K 45/06 20130101; A61P 7/12
20180101; A61P 25/28 20180101; A61P 11/02 20180101; A61K 9/2009
20130101; A61K 9/1652 20130101; A61K 31/443 20130101; A61P 1/10
20180101; A61P 5/18 20180101; A61P 9/00 20180101; A61P 13/12
20180101; A61P 1/18 20180101; A61P 5/14 20180101; A61P 19/00
20180101; A61K 9/2054 20130101; A61P 19/08 20180101; A61P 27/02
20180101; A61K 9/1682 20130101; A61K 9/2095 20130101; A61P 21/04
20180101; A61P 25/16 20180101; A61K 9/0053 20130101; A61P 1/16
20180101; A61P 15/00 20180101; A61K 9/2027 20130101; C07D 405/14
20130101; A61P 25/14 20180101; A61K 9/2013 20130101; A61P 3/06
20180101; A61P 11/00 20180101; A61P 37/02 20180101; A61P 11/06
20180101; A61K 31/47 20130101; A61K 2300/00 20130101; A61K 31/443
20130101; A61K 2300/00 20130101 |
Class at
Publication: |
424/465 ;
514/338; 424/400; 514/312 |
International
Class: |
A61K 31/443 20060101
A61K031/443; A61K 9/28 20060101 A61K009/28; A61K 45/06 20060101
A61K045/06; A61K 9/14 20060101 A61K009/14 |
Claims
1. A tablet for oral administration comprising: a. Compound 1,
Compound 1 Form I, Compound 1 Form II, and/or Compound 1 HCl Salt
Form A; b. a filler; c. a diluent; d. a disintegrant; e. a
surfactant; f. a lubricant; and g. at least one of a binder and a
glidant.
2. The tablet of claim 1, wherein Compound 1, Compound 1 Form I,
Compound 1 Form II, and/or Compound 1 HCl Salt Form A is present in
the tablet in an amount ranging from about 25 mg to about 250
mg.
3. The tablet of claim 1 or 2, wherein the amount of Compound 1,
Compound 1 Form I, Compound 1 Form II, and/or Compound 1 HCl Salt
Form A in the tablet ranges from about 15 wt % to about 75 wt % by
weight of the tablet, or from about 20 wt % to about 45 wt % by
weight of the tablet, from about 40 wt % to about 60 wt % by weight
of the tablet.
4. (canceled)
5. (canceled)
6. The tablet of claim 1, wherein the filler is selected from
cellulose, modified cellulose, sodium carboxymethyl cellulose,
ethyl cellulose hydroxymethyl cellulose, hydroxypropylcellulose,
cellulose acetate, microcrystalline cellulose, dibasic calcium
phosphate, sucrose, lactose, corn starch, potato starch, or any
combination thereof.
7. The tablet of claim 6, wherein the filler is microcrystalline
cellulose (MCC) and is present in the tablet in an amount ranging
from about 20 wt % to about 50 wt % by weight of the tablet.
8. The tablet of claim 1, wherein the diluent is selected from
lactose, mannitol, sorbitol, cellulose, calcium phosphate, starch,
sugar or any combination thereof.
9. The tablet of claim 8, wherein the diluent is mannitol and is
present in the tablet in an amount ranging from about 1 wt % to
about 30 wt % by weight of the tablet.
10. The tablet of claim 1, wherein the disintegrant is selected
from agar-agar, algins, calcium carbonate, carboxmethylcellulose,
cellulose, hydroxypropylcellulose, low substituted
hydroxypropylcellulose, clays, croscarmellose sodium, crospovidone,
gums, magnesium aluminum silicate, methylcellulose, polacrilin
potassium, sodium alginate, sodium starch glycolate, maize starch,
potato starch, tapioca starch, or any combination thereof.
11. The tablet of claim 10, wherein the disintegrant is
croscarmellose sodium and is present in the tablet at a
concentration of 5 wt % or less by weight of the tablet.
12. The tablet of claim 1, wherein the surfactant is selected from
sodium lauryl sulfate, sodium stearyl fumerate, polyoxyethylene 20
sorbitan mono-oleate, or any combination thereof.
13. The tablet of claim 12, wherein the surfactant is sodium lauryl
sulfate and has a concentration of about 5 wt % or less by weight
of the tablet.
14. The tablet of claim 1, wherein the glidant is selected from
colloidal silicon dioxide, talc, corn starch, or a combination
thereof.
15. The tablet of claim 14, wherein the glidant is colloidal
silicon dioxide and has a concentration of 5 wt % or less by weight
of the tablet.
16. The tablet of claim 1, wherein the binder is selected from
polyvinylpyrrolidone, dibasic calcium phosphate, sucrose, corn
starch, modified cellulose, or any combination thereof.
17. The tablet of claim 16, wherein the binder is
polyvinylpyrrolidone and has a concentration of less than 10 wt %
by weight of the tablet.
18. The tablet of claim 1, wherein the lubricant is selected from
magnesium stearate, calcium stearate, zinc stearate, sodium
stearate, stearic acid, aluminum stearate, leucine, glyceryl
behenate, hydrogenated vegetable oil or any combination
thereof.
19. The tablet of claim 18, wherein the lubricant is magnesium
stearate and has a concentration of less than 5 wt % by weight of
the tablet.
20. The tablet of claim 1, wherein the tablet further comprises a
colorant.
21. A pharmaceutical composition comprising a plurality of
granules, the composition comprising: a. Compound 1, Compound 1
Form I, Compound 1 Form II, and/or Compound 1 HCl Salt Form A in an
amount ranging from about 20 wt % to about 80 wt % by weight of the
composition; b. a filler in an amount ranging from about 20 wt % to
about 50 wt % by weight of the composition of a filler; c. a
disintegrant in an amount ranging from a from about 1 wt % to about
5 wt % by weight of the composition; d. a surfactant in an amount
ranging from about 2 wt % to about 0.3 wt % by weight of the
composition; e. a diluent in an amount ranging from about 1 wt % to
about 30 wt % by weight of the composition; f. a lubricant in an
amount ranging from about 0.3 wt % to about 5 wt % by weight of the
composition; and g. at least one of a binder in an amount from
about 20 wt % to about 45 wt % by weight of the composition or a
glidant in an amount ranging from about 0.05 wt % to about 2 wt %
by weight of the composition.
22. The tablet of claim 1 comprising: a. Compound 1, Compound 1
Form I, Compound 1 Form II, and/or Compound 1 HCl Salt Form A in an
amount ranging from about 25 mg to about 250 mg; b. a filler; c. a
diluent; d. a disintegrant; e. a surfactant; f. a lubricant; and g.
at least one of a binder and a glidant.
23. The tablet of claim 1 of one the following formulations:
TABLE-US-00030 (% w/w) Roller Compaction Granule Blend Compound 1
Form I 30 Microcrystalline cellulose 42.3 Mannitol 21.2
Croscarmellose Sodium 3 Sodium Lauryl Sulfate 1 Colloidal Silica
0.5 Magnesium Stearate 2 Tablet Composition (100 mg dose, 335 mg
image) Roller Compaction Granule Blend 99.5 Magnesium Stearate
0.5
TABLE-US-00031 (% w/w) High Shear Granule Blend Compound 1 Form I
50 Microcrystalline cellulose 30 Mannitol 13 Croscarmellose Sodium
2 Polyvinylpyrrolidone 4 Sodium Lauryl Sulfate 1 Water (removed
during drying) 25-40% solids Tablet Composition (100 mg dose, 205
mg image) High Shear Granule Blend 97.5 Croscarmellose Sodium 2.0
Magnesium Stearate 0.5
TABLE-US-00032 (% w/w) High Shear Granule Blend Compound 1 Form I
60 Microcrystalline cellulose 20 Mannitol 13 Croscarmellose Sodium
2 Polyvinylpyrrolidone 4 Sodium Lauryl Sulfate 1 Water (removed
during drying) 25-40% solids Tablet Composition (100 mg dose, 171
mg image) High Shear Granule Blend 97.5 Croscarmellose Sodium 2.0
Magnesium Stearate 0.5
TABLE-US-00033 (% w/w) High Shear Granule Blend Compound 1 Form I
60 Microcrystalline cellulose 20 Mannitol 13 Croscarmellose Sodium
2 Polyvinylpyrrolidone 4 Sodium Lauryl Sulfate 1 Water (removed
during drying) 25-40% solids Tablet Composition (200 mg dose, 402
mg image) High Shear Granule Blend 83 Microcrystalline cellulose 14
Croscarmellose Sodium 2 Magnesium Stearate 1.0
TABLE-US-00034 mg High Shear Granule Blend Compound 1 Form I 200
Microcrystalline cellulose 66 Mannitol 43 Croscarmellose Sodium 7
Polyvinylpyrrolidone 13 Sodium Lauryl Sulfate 3 Core Tablet
Composition (200 mg dose, 400 mg image) High Shear Granule Blend
332 Microcrystalline cellulose 56 Croscarmellose Sodium 8 Magnesium
Stearate 4 Film Coated Tablet (200 mg dose, 412 mg image) Core
Tablet Composition 400 Film Coat 12 Wax 0.04
TABLE-US-00035 mg High Shear Granule Blend Compound 1 Form I 200
Microcrystalline cellulose 67 Mannitol 45 Croscarmellose Sodium 7
Polyvinylpyrrolidone 10.4 Sodium Lauryl Sulfate 2.6 Core Tablet
Composition (200 mg dose, 400 mg image) High Shear Granule Blend
332 Microcrystalline cellulose 56 Croscarmellose Sodium 8 Magnesium
Stearate 4 Film Coated Tablet (200 mg dose, 412 mg image) Core
Tablet Composition 400 Film Coat 12 Wax 0.04
TABLE-US-00036 (% w/w) High Shear Granule Blend Compound 1 Form I
70 Microcrystalline cellulose 12 Mannitol 11 Croscarmellose Sodium
2 Polyvinylpyrrolidone 4 Sodium Lauryl Sulfate 1 Water (removed
during drying) 25-40% solids Tablet Composition (100 mg dose, 147
mg image) High Shear Granule Blend 97.5 Croscarmellose Sodium 2.0
Magnesium Stearate 0.5
TABLE-US-00037 (% w/w) High Shear Granule Blend Compound 1 Form I
or Form II 61 Microcrystalline cellulose 20.3 Mannitol 13.2
Croscarmellose Sodium 2 Polyvinylpyrrolidone 2.7 Sodium Lauryl
Sulfate 0.7 Tablet Composition (100 mg dose, 197 mg image) High
Shear Granule Blend 83 Microcrystalline cellulose 14 Croscarmellose
Sodium 2 Magnesium Stearate 1
TABLE-US-00038 mg High Shear Granule Blend Compound 1 Form I or
Form II 100 Microcrystalline cellulose 33.3 Mannitol 21.7
Croscarmellose Sodium 3.3 Polyvinylpyrrolidone 4.4 Sodium Lauryl
Sulfate 1.1 Core Tablet Composition (100 mg dose, 197 mg image)
High Shear Granule Blend 163.9 Microcrystalline cellulose 27.6
Croscarmellose Sodium 3.9 Magnesium Stearate 2.0
24. (canceled)
25. (canceled)
26. (canceled)
27. (canceled)
28. (canceled)
29. (canceled)
30. (canceled)
31. (canceled)
32. The pharmaceutical composition of claim 21 which is suitable
for oral administration comprising: a. Compound 1, Compound 1 Form
I, Compound 1 Form II, and/or Compound 1 HCl Salt Form A in an
amount ranging from about 20 wt % to about 80 wt % by weight of the
composition; b. a filler comprising microcrystalline cellulose in
an amount ranging from about 20 wt % to about 50 wt % by weight of
the composition; c. a disintegrant comprising sodium croscarmellose
sodium in an amount ranging from about 1 wt % to about 5 wt % by
weight of the composition; d. a surfactant comprising sodium lauryl
sulfate in an amount ranging from about 2 wt % to about 0.3 wt % by
weight of the composition; e. a diluent comprising mannitol in an
amount ranging from about 1 wt % to about 30 wt % by weight of the
composition; f. a lubricant comprising magnesium stearate in an
amount ranging from about 0.3 wt % to about 5 wt % by weight of the
composition; and g. at least one of: a binder comprising
polyvinylpyrrolidone in an amount ranging from about 0.1 wt % to
about 5 wt % by weight of the composition and a glidant comprising
colloidal silica in an amount ranging from about 0.05 wt % to about
2 wt % by weight of the composition.
33. The pharmaceutical composition of claim 32, further comprising
about 0.4 wt % of colorant by weight of the composition.
34. The pharmaceutical composition of claim 32, wherein the
pharmaceutical composition comprises a plurality of granules,
wherein the plurality of granules have a mean or average particle
diameter ranging from 100 .mu.m to about 2 mm.
35. (canceled)
36. The pharmaceutical composition of claim 32, wherein the
pharmaceutical composition is a tablet comprising a coating.
37. (canceled)
38. The pharmaceutical composition of claim 32, wherein the
pharmaceutical composition further comprises at least one
additional therapeutic agent, wherein the additional therapeutic
agent is a CFTR modulator or a CFTR potentiator.
39. (canceled)
40. (canceled)
41. A dosage unit form which comprises: a. about 30 wt % of
Compound 1, Compound 1 Form I, Compound 1 Form II, and/or Compound
1 HCl Salt Form A by weight of the composition; b. about 42 wt % of
microcrystalline cellulose by weight of the composition; c. about
21 wt % of mannitol by weight of the composition; d. about 3 wt %
of sodium croscarmellose sodium by weight of the composition; e.
about 1 wt % of sodium lauryl sulfate by weight of the composition;
f. about 2.5 wt % of magnesium stearate by weight of the
composition; and g. about 0.5 wt % of colloidal silica by weight of
the composition; a. about 50 wt % of Compound 1, Compound 1 Form I,
Compound 1 Form II, and/or Compound 1 HCl Salt Form A; b. about 30
wt % of microcrystalline cellulose by weight of the composition; c.
about 13 wt % of mannitol by weight of the composition; d. about 2
wt % of sodium croscarmellose sodium by weight of the composition;
e. about 4 wt % of polyvinylpyrrolidone by weight of the
composition; f. about 1 wt % of sodium lauryl sulfate by weight of
the composition; and g. about 0.5 wt % of magnesium stearate by
weight of the composition; a. about 60 wt % of Compound 1, Compound
1 Form I, Compound 1 Form II, and/or Compound 1 HCl Salt Form A; b.
about 20 wt % of microcrystalline cellulose by weight of the
composition; c. about 13 wt % of mannitol by weight of the
composition; d. about 2 wt % of sodium croscarmellose sodium by
weight of the composition; e. about 4 wt % of polyvinylpyrrolidone
by weight of the composition; f. about 1 wt % of sodium lauryl
sulfate by weight of the composition; and g. about 0.5 wt % of
magnesium stearate by weight of the composition; a. about 60 wt %
of Compound 1, Compound 1 Form I, Compound 1 Form II, and/or
Compound 1 HCl Salt Form A; b. about 34 wt % of microcrystalline
cellulose by weight of the composition; c. about 13 wt % of
mannitol by weight of the composition; d. about 4 wt % of sodium
croscarmellose sodium by weight of the composition; e. about 4 wt %
of polyvinylpyrrolidone by weight of the composition; f. about 1 wt
% of sodium lauryl sulfate by weight of the composition; and g.
about 1.5 wt % of magnesium stearate by weight of the composition;
a. about 150 to 250 mg of Compound 1, Compound 1 Form I, Compound 1
Form II, and/or Compound 1 HCl Salt Form A; b. about 40 to 50 mg of
mannitol; c. about 120 to 130 mg of microcrystalline cellulose; d.
about 10 to 20 mg of croscarmellose sodium; e. about 10 to 20 mg of
polyvinylpyrrolidone; f. about 1 to 5 mg of sodium lauryl sulfate;
and g. about 1 to 5 mg of magnesium stearate; a. about 200 mg of
Compound 1, Compound 1 Form I, Compound 1 Form II, and/or Compound
1 HCl Salt Form A; b. about 43 mg of mannitol; c. about 123 mg of
microcrystalline cellulose; d. about 15 mg of croscarmellose
sodium; e. about 13 mg of polyvinylpyrrolidone; f. about 3 mg of
sodium lauryl sulfate; and g. about 4 mg of magnesium stearate; or
a. about 70 wt % of Compound 1, Compound 1 Form I, Compound 1 Form
II, and/or Compound 1 HCl Salt Form A; b. about 12 wt % of
microcrystalline cellulose by weight of the composition; c. about
11 wt % of mannitol by weight of the composition; d. about 2 wt %
of sodium croscarmellose sodium by weight of the composition; e.
about 4 wt % of polyvinylpyrrolidone by weight of the composition;
f. about 1 wt % of sodium lauryl sulfate by weight of the
composition; and g. about 0.5 wt % of magnesium stearate by weight
of the composition.
42. (canceled)
43. (canceled)
44. (canceled)
45. (canceled)
46. (canceled)
47. (canceled)
48. The tablet, pharmaceutical composition, or dosage unit form of
claim 1, 21, or 41, wherein Compound 1, Compound 1 Form I, Compound
1 Form II, and/or Compound 1 HCl Salt Form A has a particle size of
0.1 microns to 50 microns, 0.1 to 20 microns, 0.1 to 10 microns,
1.0 to 5 microns, or a particle size D50 of 2.0 microns.
49. (canceled)
50. (canceled)
51. (canceled)
52. (canceled)
53. A method of producing a pharmaceutical composition comprising
the steps of: combining a therapeutically effective amount of
Compound 1, Compound 1 Form I, Compound 1 Form II, and/or Compound
1 HCl Salt Form A and at least one granulation excipient selected
from the group consisting of: a binder; a glidant; a surfactant; a
lubricant; a disintegrant; a filler, a diluent and combinations
thereof to form an admixture; mixing the admixture; and compacting
the admixture to form the pharmaceutical composition.
54. The method of claim 53, wherein the pharmaceutical composition
comprises a plurality of granules.
55. The method of claim 53, wherein compacting the admixture
comprises compacting the admixture in a roller compactor forming
compressed sheets of admixture; and milling the sheets of admixture
to form a plurality of granules.
56. The method of claim 54, further comprising compressing the
plurality of granules with at least one pharmaceutical acceptable
excipient to form a tablet, wherein the at least one pharmaceutical
acceptable excipient is selected from the group consisting of
magnesium stearate, croscarmellose sodium and combinations
thereof.
57. (canceled)
58. The method according to claim 54, wherein the plurality of
granules are compressed to produce a tablet having a hardness of at
least 5 kP.
59. The method of claim 53, wherein the step of compacting the
admixture to form the pharmaceutical composition further comprises
drying the admixture.
60. The method of claim 53, wherein mixing the admixture comprises
mixing the admixture until the admixture is substantially
homogenous.
61. The method of claim 53, wherein a plurality of granules are
formed by combining Compound 1, Compound 1 Form I, Compound 1 Form
II, and/or Compound 1 HCl Salt Form A with a granulation fluid
comprising a surfactant and a binder.
62. The method of claim 61, wherein the surfactant is sodium lauryl
sulfate.
63. A method of administering a tablet comprising orally
administering to a patient at least once per day a tablet
comprising: a. about 25 to 200 mg of Compound 1, Compound 1 Form I,
Compound 1 Form II, and/or Compound 1 HCl Salt Form A; b. a filler;
c. a diluent; d. a disintegrant; e. a surfactant; f. at least one
of a binder and a glidant; and g. a lubricant.
64. The method of claim 63, wherein the tablet comprises about 25
mg, about 50 mg, about 75 mg, about 100 mg, about 150 mg, or about
200 mg of Compound 1, Compound 1 Form I, Compound 1 Form II, and/or
Compound 1 HCl Salt Form A.
65. (canceled)
66. (canceled)
67. (canceled)
68. (canceled)
69. The method of claim 63, wherein the tablet is orally
administered to the patient twice per day.
70. The method of claim 69, wherein the tablet comprises about 25
mg about 50 mg, about 75 mg, about 100 mg, about 150 mg, or about
200 mg of Compound 1, Compound 1 Form I, Compound 1 Form II, and/or
Compound 1 HCl Salt Form A.
71. (canceled)
72. (canceled)
73. (canceled)
74. (canceled)
75. (canceled)
76. The method of claim 63, wherein the tablet is orally
administered to the patient once every 12 hours.
77. The method of claim 76, wherein the tablet comprises about 25
mg about 50 mg, about 75 mg, about 100 mg, about 150 mg, or about
200 mg of Compound 1, Compound 1 Form I, Compound 1 Form II, and/or
Compound 1 HCl Salt Form A.
78. (canceled)
79. (canceled)
80. (canceled)
81. (canceled)
82. (canceled)
83. A method of treating or lessening the severity of a disease in
a subject comprising administering to the subject a tablet,
pharmaceutical composition, or dosage unit form of Compound 1,
Compound 1 Form I, Compound 1 Form II, and/or Compound 1 HCl Salt
Form A, wherein the disease is selected from cystic fibrosis,
asthma, smoke induced COPD, chronic bronchitis, rhinosinusitis,
constipation, pancreatitis, pancreatic insufficiency, male
infertility caused by congenital bilateral absence of the vas
deferens (CBAVD), mild pulmonary disease, idiopathic pancreatitis,
allergic bronchopulmonary aspergillosis (ABPA), liver disease,
hereditary emphysema, hereditary hemochromatosis,
coagulation-fibrinolysis deficiencies, protein C deficiency, Type 1
hereditary angioedema, lipid processing deficiencies, familial
hypercholesterolemia, Type 1 chylomicronemia, abetalipoproteinemia,
lysosomal storage diseases, I-cell disease/pseudo-Hurler,
mucopolysaccharidoses, Sandhof/Tay-Sachs, Crigler-Najjar type II,
polyendocrinopathy/hyperinsulemia, Diabetes mellitus, Laron
dwarfism, myleoperoxidase deficiency, primary hypoparathyroidism,
melanoma, glycanosis CDG type 1, congenital hyperthyroidism,
osteogenesis imperfecta, hereditary hypofibrinogenemia, ACT
deficiency, Diabetes insipidus (DI), neurophyseal DI, neprogenic
DI, Charcot-Marie Tooth syndrome, Perlizaeus-Merzbacher disease,
neurodegenerative diseases, Alzheimer's disease, Parkinson's
disease, amyotrophic lateral sclerosis, progressive supranuclear
plasy, Pick's disease, several polyglutamine neurological
disorders, Huntington's, spinocerebullar ataxia type I, spinal and
bulbar muscular atrophy, dentatorubal pallidoluysian, myotonic
dystrophy, spongiform encephalopathies, hereditary
Creutzfeldt-Jakob disease (due to prion protein processing defect),
Fabry disease, Straussler-Scheinker syndrome, COPD, dry-eye
disease, Sjogren's disease, Osteoporosis, Osteopenia, Gorham's
Syndrome, chloride channelopathies, myotonia congenita (Thomson and
Becker forms), Bartter's syndrome type III, Dent's disease,
hyperekplexia, epilepsy, hyperekplexia, lysosomal storage disease,
Angelman syndrome, Primary Ciliary Dyskinesia (PCD), inherited
disorders of the structure and/or function of cilia, PCD with situs
inversus (also known as Kartagener syndrome), PCD without situs
inversus, or ciliary aplasia.
84. The method of claim 83, wherein the disease is disease is
cystic fibrosis, emphysema, COPD, or osteoporosis.
85. The method of claim 83, wherein the disease is cystic
fibrosis.
86. The method of claim 83, wherein said patient has cystic
fibrosis transmembrane receptor (CFTR) with a .DELTA.F508 mutation,
a R117H mutation, or a G551D mutation.
87. (canceled)
88. (canceled)
89. The method of claim 83, wherein the method comprises
administering an additional therapeutic agent.
90. A kit comprising a tablet, pharmaceutical composition, or
dosage unit form of Compound 1, Compound 1 Form I, Compound 1 Form
II, and/or Compound 1 HCl Salt Form A and instructions for use
thereof.
91. The kit of claim 90, wherein Compound 1 is Compound 1 Form I
and is present in the tablet, pharmaceutical composition, or dosage
unit in an amount ranging from 25 mg to about 200 mg.
92. The pharmaceutical composition of claim 38, wherein the CFTR
modulator is
N-(5-hydroxy-2,4-ditert-butyl-phenyl)-4-oxo-1H-quinoline-3-carboxamide.
93. The method of claim 89 wherein the additional therapeutic agent
is
N-(5-hydroxy-2,4-ditert-butyl-phenyl)-4-oxo-1H-quinoline-3-carboxamide.
Description
TECHNICAL FIELD OF THE INVENTION
[0001] The invention relates to pharmaceutical compositions
comprising
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid (Compound 1), methods for
manufacturing such compositions and methods for administering
pharmaceutical compositions comprising same.
BACKGROUND
[0002] CFTR is a cAMP/ATP-mediated anion channel that is expressed
in a variety of cells types, including absorptive and secretory
epithelia cells, where it regulates anion flux across the membrane,
as well as the activity of other ion channels and proteins. In
epithelia cells, normal functioning of CFTR is critical for the
maintenance of electrolyte transport throughout the body, including
respiratory and digestive tissue. CFTR is composed of approximately
1480 amino acids that encode a protein made up of a tandem repeat
of transmembrane domains, each containing six transmembrane helices
and a nucleotide binding domain. The two transmembrane domains are
linked by a large, polar, regulatory (R)-domain with multiple
phosphorylation sites that regulate channel activity and cellular
trafficking.
[0003] The gene encoding CFTR has been identified and sequenced
(See Gregory, R. J. et al. (1990) Nature 347:382-386; Rich, D. P.
et al. (1990) Nature 347:358-362), (Riordan, J. R. et al. (1989)
Science 245:1066-1073). A defect in this gene causes mutations in
CFTR resulting in cystic fibrosis ("CF"), the most common fatal
genetic disease in humans. Cystic fibrosis affects approximately
one in every 2,500 infants in the United States. Within the general
United States population, up to 10 million people carry a single
copy of the defective gene without apparent ill effects. In
contrast, individuals with two copies of the CF associated gene
suffer from the debilitating and fatal effects of CF, including
chronic lung disease.
[0004] In patients with cystic fibrosis, mutations in CFTR
endogenously expressed in respiratory epithelia lead to reduced
apical anion secretion causing an imbalance in ion and fluid
transport. The resulting decrease in anion transport contributes to
enhance mucus accumulation in the lung and the accompanying
microbial infections that ultimately cause death in CF patients. In
addition to respiratory disease, CF patients typically suffer from
gastrointestinal problems and pancreatic insufficiency that, if
left untreated, results in death. In addition, the majority of
males with cystic fibrosis are infertile and fertility is decreased
among females with cystic fibrosis. In contrast to the severe
effects of two copies of the CF associated gene, individuals with a
single copy of the CF associated gene exhibit increased resistance
to cholera and to dehydration resulting from diarrhea--perhaps
explaining the relatively high frequency of the CF gene within the
population.
[0005] Sequence analysis of the CFTR gene of CF chromosomes has
revealed a variety of disease-causing mutations (Cutting, G. R. et
al. (1990) Nature 346:366-369; Dean, M. et al. (1990) Cell
61:863:870; and Kerem, B-S. et al. (1989) Science 245:1073-1080;
Kerem, B-S et al. (1990) Proc. Natl. Acad. Sci. USA 87:8447-8451).
To date, greater than 1000 disease-causing mutations in the CF gene
have been identified as reported by the scientific and medical
literature. The most prevalent mutation is a deletion of
phenylalanine at position 508 of the CFTR amino acid sequence, and
is commonly referred to as .DELTA.F508-CFTR. This mutation occurs
in approximately 70 percent of the cases of cystic fibrosis and is
associated with a severe disease. Other mutations include the R117H
and G551D.
[0006] The deletion of residue 508 in .DELTA.F508-CFTR prevents the
nascent protein from folding correctly. This results in the
inability of the mutant protein to exit the ER, and traffic to the
plasma membrane. As a result, the number of channels present in the
membrane is far less than observed in cells expressing wild-type
CFTR. In addition to impaired trafficking, the mutation results in
defective channel gating. Together, the reduced number of channels
in the membrane and the defective gating lead to reduced anion
transport across epithelia leading to defective ion and fluid
transport. (Quinton, P. M. (1990), FASEB J. 4: 2709-2727). Studies
have shown, however, that the reduced numbers of .DELTA.F508-CFTR
in the membrane are functional, albeit less than wild-type CFTR.
(Dalemans et al. (1991), Nature Lond. 354: 526-528; Denning et al.,
supra; Pasyk and Foskett (1995), J. Cell. Biochem. 270: 12347-50).
In addition to .DELTA.F508-CFTR, other disease causing mutations in
CFTR that result in defective trafficking, synthesis, and/or
channel gating could be up- or down-regulated to alter anion
secretion and modify disease progression and/or severity.
[0007] Although CFTR transports a variety of molecules in addition
to anions, it is clear that this role (the transport of anions)
represents one element in an important mechanism of transporting
ions and water across the epithelium. The other elements include
the epithelial Na.sup.+ channel, ENaC, Na.sup.+/2Cl.sup.-/K.sup.+
co-transporter, Na.sup.+--K.sup.+-ATPase pump and the basolateral
membrane K.sup.+ channels, that are responsible for the uptake of
chloride into the cell.
[0008] These elements work together to achieve directional
transport across the epithelium via their selective expression and
localization within the cell. Chloride absorption takes place by
the coordinated activity of ENaC and CFTR present on the apical
membrane and the Na.sup.+--K.sup.+-ATPase pump and Cl-- channels
expressed on the basolateral surface of the cell. Secondary active
transport of chloride from the luminal side leads to the
accumulation of intracellular chloride, which can then passively
leave the cell via Cl.sup.- channels, resulting in a vectorial
transport. Arrangement of Na.sup.+/2Cl.sup.-/K.sup.+
co-transporter, Na.sup.+--K.sup.+-ATPase pump and the basolateral
membrane K.sup.+ channels on the basolateral surface and CFTR on
the luminal side coordinate the secretion of chloride via CFTR on
the luminal side. Because water is probably never actively
transported itself, its flow across epithelia depends on tiny
transepithelial osmotic gradients generated by the bulk flow of
sodium and chloride.
[0009] As discussed above, it is believed that the deletion of
residue 508 in .DELTA.F508-CFTR prevents the nascent protein from
folding correctly, resulting in the inability of this mutant
protein to exit the ER, and traffic to the plasma membrane. As a
result, insufficient amounts of the mature protein are present at
the plasma membrane and chloride transport within epithelial
tissues is significantly reduced. In fact, this cellular phenomenon
of defective endoplasmic reticulum (ER) processing of ATP-binding
cassette (ABC) transporters by the ER machinery, has been shown to
be the underlying basis not only for CF disease, but for a wide
range of other isolated and inherited diseases. The two ways that
the ER machinery can malfunction is either by loss of coupling to
ER export of the proteins leading to degradation, or by the ER
accumulation of these defective/misfolded proteins [Aridor M, et
al., Nature Med., 5(7), pp 745-751 (1999); Shastry, B. S., et al.,
Neurochem. International, 43, pp 1-7 (2003); Rutishauser, J., et
al., Swiss Med Wkly, 132, pp 211-222 (2002); Morello, J P et al.,
TIPS, 21, pp. 466-469 (2000); Bross P., et al., Human Mut., 14, pp.
186-198 (1999)].
[0010]
3-(6-(1-(2,2-Difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxami-
do)-3-methylpyridin-2-yl)benzoic acid in salt form is disclosed in
International PCT Publication WO 2007056341 as a modulator of CFTR
activity and thus as a useful treatment for CFTR-mediated diseases
such as cystic fibrosis. Form I of
3-(6-(1-(2,2-Difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid, which is a substantially
crystalline and salt-free form known as Compound 1 Form I, is
disclosed in U.S. patent application Ser. No. 12/327,902, filed
Dec. 4, 2008. Form II and HCl salt Form A of
3-(6(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarbox-
amido)-3-methylpyridin-2-yl)benzoic acid, Compound 1 Form II and
Compound 1 HCl salt Form A, respectively, are disclosed in U.S.
Provisional Patent Application 61/321,729, filed Apr. 7, 2010. All
applications are incorporated in their entirety by reference
herein. A need remains, however, for pharmaceutical compositions
comprising Compound 1 Form I, Form II, or HCl salt Form A that are
readily prepared and that are suitable for use as therapeutics.
SUMMARY
[0011] The invention relates to pharmaceutical compositions,
pharmaceutical preparations, and solid dosage forms comprising
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid (Compound 1) which has the
structure below:
##STR00001##
[0012] In one aspect, the invention provides a pharmaceutical
composition comprising:
[0013] a. Compound 1;
[0014] b. a filler;
[0015] c. a disintegrant;
[0016] d. a surfactant;
[0017] e. a diluent;
[0018] f. a lubricant; and
[0019] g. at least one of a glidant and a binder.
[0020] In other embodiments, Compound 1 is in substantially one of
its crystalline solid forms. In one embodiment, Compound 1 is in
substantially crystalline Form I (Compound 1 Form I). In one
embodiment, Compound 1 is in substantially crystalline Form II
(Compound 1 Form II). In one embodiment, Compound 1 is in
substantially crystalline HCl salt form (Compound 1 HCl Salt Form
A). It is understood that the term "Compound 1," as used
throughout, includes, amongst other forms, including
non-crystalline forms, the following solid state forms: Compound 1
Form I, Compound 1 Form II, and/or Compound 1 HCl Salt Form A.
[0021] In some embodiments, the pharmaceutical composition
comprises 25 mg to 400 mg. In some embodiments, the pharmaceutical
composition comprises 25 mg of Compound 1. In some embodiments, the
pharmaceutical composition comprises 50 mg of Compound 1. In some
embodiments, the pharmaceutical composition comprises 100 mg of
Compound 1. In some embodiments, the pharmaceutical composition
comprises 125 mg of Compound 1. In some embodiments, the
pharmaceutical composition comprises 150 mg of Compound 1. In some
embodiments, the pharmaceutical composition comprises 200 mg of
Compound 1. In some embodiments, the pharmaceutical composition
comprises 250 mg of Compound 1. In some embodiments, the
pharmaceutical composition comprises 400 mg of Compound 1.
[0022] In one aspect, the invention provides a pharmaceutical
composition comprising the following components:
TABLE-US-00001 (% w/w) Roller Compaction Granule Blend Compound 1
20-40 Microcrystalline cellulose 30-50 Mannitol 10-30
Croscarmellose Sodium 1-5 Sodium Lauryl Sulfate 0.1-2 Colloidal
Silica 0.1-1 Magnesium Stearate 1-3 Tablet Composition (100 mg
dose) Roller Compaction Granule Blend 99-99.9 Magnesium Stearate
0.1-1
[0023] In another embodiment, the invention provides a
pharmaceutical composition comprising the following components:
TABLE-US-00002 (% w/w) Roller Compaction Granule Blend Compound 1
Form I 30 Microcrystalline cellulose 42.3 Mannitol 21.2
Croscarmellose Sodium 3 Sodium Lauryl Sulfate 1 Colloidal Silica
0.5 Magnesium Stearate 2 Tablet Composition (100 mg dose, 335 mg
image) Roller Compaction Granule Blend 99.5 Magnesium Stearate
0.5
[0024] In another aspect, the invention provides a pharmaceutical
composition comprising the following components:
TABLE-US-00003 (% w/w) High Shear Granule Blend Compound 1 Form I
40-80 Microcrystalline cellulose 20-40 Mannitol 10-15
Croscarmellose Sodium 1-5 Polyvinylpyrrolidone 1-10 Sodium Lauryl
Sulfate 0.1-2 Water (removed during drying) 25-40% solids Tablet
Composition (100 mg dose) High Shear Granule Blend 95-99
Croscarmellose Sodium 1-4 Magnesium Stearate 0.1-1
[0025] In another embodiment, the invention provides a
pharmaceutical composition comprising the following components:
TABLE-US-00004 (% w/w) High Shear Granule Blend Compound 1 Form I
50 Microcrystalline cellulose 30 Mannitol 13 Croscarmellose Sodium
2 Polyvinylpyrrolidone 4 Sodium Lauryl Sulfate 1 Water (removed
during drying) 25-40% solids Tablet Composition (100 mg dose, 205
mg image) High Shear Granule Blend 97.5 Croscarmellose Sodium 2.0
Magnesium Stearate 0.5
[0026] In another embodiment, the invention provides a
pharmaceutical composition comprising the following components:
TABLE-US-00005 (% w/w) High Shear Granule Blend Compound 1 Form I
60 Microcrystalline cellulose 20 Mannitol 13 Croscarmellose Sodium
2 Polyvinylpyrrolidone 4 Sodium Lauryl Sulfate 1 Water (removed
during drying) 25-40% solids Tablet Composition (100 mg dose, 171
mg image) High Shear Granule Blend 97.5 Croscarmellose Sodium 2.0
Magnesium Stearate 0.5
[0027] In another embodiment, the invention provides a
pharmaceutical composition comprising the following components:
TABLE-US-00006 (% w/w) High Shear Granule Blend Compound 1 Form I
60 Microcrystalline cellulose 20 Mannitol 13 Croscarmellose Sodium
2 Polyvinylpyrrolidone 4 Sodium Lauryl Sulfate 1 Water (removed
during drying) 25-40% solids Tablet Composition (200 mg dose, 402
mg image) High Shear Granule Blend 83 Microcrystalline cellulose 14
Croscarmellose Sodium 2 Magnesium Stearate 1
[0028] In another embodiment, the invention provides a
pharmaceutical composition comprising the following components:
TABLE-US-00007 mg High Shear Granule Blend Compound 1 Form I 200
Microcrystalline cellulose 66 Mannitol 43 Croscarmellose Sodium 7
Polyvinylpyrrolidone 13 Sodium Lauryl Sulfate 3 Core Tablet
Composition (200 mg dose, 400 mg image) High Shear Granule Blend
332 Microcrystalline cellulose 56 Croscarmellose Sodium 8 Magnesium
Stearate 4 Film Coated Tablet (200 mg dose, 412 mg image) Core
Tablet Composition 400 Film Coat 12 Wax 0.04
[0029] In another embodiment, the invention provides a
pharmaceutical composition comprising the following components:
TABLE-US-00008 mg High Shear Granule Blend Compound 1 Form I 200
Microcrystalline cellulose 67 Mannitol 45 Croscarmellose Sodium 7
Polyvinylpyrrolidone 10.4 Sodium Lauryl Sulfate 2.6 Core Tablet
Composition (200 mg dose, 400 mg image) High Shear Granule Blend
332 Microcrystalline cellulose 56 Croscarmellose Sodium 8 Magnesium
Stearate 4 Film Coated Tablet (200 mg dose, 412 mg image) Core
Tablet Composition 400 Film Coat 12 Wax 0.04
[0030] In another embodiment, the invention provides a
pharmaceutical composition comprising the following components:
TABLE-US-00009 (% w/w) High Shear Granule Blend Compound 1 Form I
70 Microcrystalline cellulose 12 Mannitol 11 Croscarmellose Sodium
2 Polyvinylpyrrolidone 4 Sodium Lauryl Sulfate 1 Water (removed
during drying) 25-40% solids Tablet Composition (100 mg dose, 147
mg image) High Shear Granule Blend 97.5 Croscarmellose Sodium 2.0
Magnesium Stearate 0.5
[0031] In another embodiment, the invention provides a
pharmaceutical composition comprising the following components:
TABLE-US-00010 (% w/w) High Shear Granule Blend Compound 1 Form I
or Form II 61 Microcrystalline cellulose 20.3 Mannitol 13.2
Croscarmellose Sodium 2 Polyvinylpyrrolidone 2.7 Sodium Lauryl
Sulfate 0.7 Tablet Composition (100 mg dose, 197 mg image) High
Shear Granule Blend 83 Microcrystalline cellulose 14 Croscarmellose
Sodium 2 Magnesium Stearate 1
[0032] In another embodiment, the invention provides a
pharmaceutical composition comprising the following components:
TABLE-US-00011 mg High Shear Granule Blend Compound 1 Form I or
Form II 100 Microcrystalline cellulose 33.3 Mannitol 21.7
Croscarmellose Sodium 3.3 Polyvinylpyrrolidone 4.4 Sodium Lauryl
Sulfate 1.1 Core Tablet Composition (100 mg dose, 197 mg image)
High Shear Granule Blend 163.9 Microcrystalline cellulose 27.6
Croscarmellose Sodium 3.9 Magnesium Stearate 2.0
[0033] In another aspect, the invention provides a pharmaceutical
composition in the form of a tablet that comprises Compound 1, and
one or more pharmaceutically acceptable excipients, for example, a
filler, a disintegrant, a surfactant, a diluent, a binder, a
glidant, and a lubricant and any combination thereof, where the
tablet has a dissolution of at least about 50% in about 30 minutes.
In another embodiment, the dissolution rate is at least about 75%
in about 30 minutes. In another embodiment, the dissolution rate is
at least about 90% in about 30 minutes.
[0034] In another aspect, the invention provides a pharmaceutical
composition consisting of a tablet that comprises a powder blend or
granules comprising Compound 1; and, one or more pharmaceutically
acceptable excipients, for example, a filler, a disintegrant, a
surfactant, a diluent, a binder, a glidant, and a lubricant,
wherein the tablet has a hardness of at least about 5 kP (kP=kilo
Ponds; 1 kP==.about.9.8 N). In another embodiment, the tablet has a
target friability of less than 1.0% after 400 revolutions. In
another aspect, the invention provides a pharmaceutical composition
consisting of a tablet that comprises a powder blend or granules
comprising Compound 1 Form II, Compound 1; and, one or more
pharmaceutically acceptable excipients, for example, a filler, a
disintegrant, a surfactant, a diluent, a binder, a glidant, and a
lubricant, wherein the tablet has a hardness of at least about 5 kP
(kP=kilo Ponds; 1 kP=.about.9.8 N). In another embodiment, the
tablet has a target friability of less than 1.0% after 400
revolutions.
[0035] In another aspect, the invention provides a pharmaceutical
composition as described herein further comprising an additional
therapeutic agent. In some embodiments, the additional therapeutic
agent is
N-(5-hydroxy-2,4-ditert-butyl-phenyl)-4-oxo-1H-quinoline-3-carboxamide-
.
[0036] In another aspect, the invention provides a method of
treating a CFTR mediated disease in a mammal comprising
administering to the mammal an effective amount of a pharmaceutical
composition as described herein. In some embodiments, the CFTR
mediated disease is cystic fibrosis, emphysema, COPD, or
osteoporosis. In other embodiments, the CFTR mediated disease is
cystic fibrosis. This method may further comprise administering an
additional therapeutic agent, wherein in some embodiments, the
additional therapeutic agent is
N-(5-hydroxy-2,4-ditert-butyl-phenyl)-4-oxo-1H-quinoline-3-carboxamide.
[0037] In another aspect, the invention provides a process for
making the pharmaceutical compositions described herein by a roller
compaction process comprising the steps of screening and weighing
Compound 1 and excipients; blending Compound 1 and excipients for a
suitable amount of time; roller compacting the blend into ribbons
and milling the ribbons into granules; blending the granules with
extra-granular excipients for a suitable amount of time;
compressing the blend into tablets; coating the tablets; and,
optionally, printing a monogram on one or both tablet faces.
[0038] In another aspect, the invention provides a process for
making the pharmaceutical compositions described herein by a high
shear granulation process comprising the steps of screening and
weighing Compound 1 and excipients; mixing Compound 1 and
excipients while adding a granulation fluid comprising surfactant
and a binder at a suitable mixing speed for a suitable amount of
time and chopping the mixture into granules; drying the granules;
blending the granules with extra-granular excipients for a suitable
amount of time; compressing the blend into tablets; coating the
tablets; and, optionally, printing a monogram on one or both tablet
faces.
BRIEF DESCRIPTION OF DRAWINGS
[0039] FIG. 1 is an X-ray diffraction pattern calculated from a
single crystal structure of Compound 1 Form I.
[0040] FIG. 2 is an actual X-ray powder diffraction pattern of
Compound 1 Form I.
[0041] FIG. 3 is an X-ray powder diffraction pattern of Compound 1
Form II.
[0042] FIG. 4 provides X-ray diffraction patterns of Compound 1
Form II's selected from:
[0043] 1) Compound 1 Form II, Methanol Solvate;
[0044] 2) Compound 1 Form II, Ethanol Solvate;
[0045] 3) Compound 1 Form II, Acetone Solvate;
[0046] 4) Compound 1 Form II, 2-Propanol Solvate;
[0047] 5) Compound 1 Form II, Acetonitrile Solvate;
[0048] 6) Compound 1 Form II, Tetrahydrofuran Solvate;
[0049] 7) Compound 1 Form II, Methyl Acetate Solvate;
[0050] 8) Compound 1 Form II, 2-Butanone Solvate;
[0051] 9) Compound 1 Form II, Ethyl Formate Solvate; and
[0052] 10) Compound 1 Form II, 2-Methyltetrahydrofuran Solvate.
[0053] FIG. 5 provides an X-ray diffraction pattern of Compound 1
Form II, Methanol Solvate.
[0054] FIG. 6 provides an X-ray diffraction pattern of Compound 1
Form II, Ethanol Solvate.
[0055] FIG. 7 provides an X-ray diffraction pattern of Compound 1
Form II, Acetone Solvate.
[0056] FIG. 8 provides an X-ray diffraction pattern of Compound 1
Form II, 2-Propanol Solvate.
[0057] FIG. 9 provides an X-ray diffraction pattern of Compound 1
Form II, Acetonitrile Solvate.
[0058] FIG. 10 provides an X-ray diffraction pattern of Compound 1
Form II, Tetrahydrofuran Solvate.
[0059] FIG. 11 provides an X-ray diffraction pattern of Compound 1
Form II, Methyl Acetate Solvate.
[0060] FIG. 12 provides an X-ray diffraction pattern of Compound 1
Form II, 2-Butanone Solvate.
[0061] FIG. 13 provides an X-ray diffraction pattern of Compound 1
Form II, Ethyl Formate Solvate.
[0062] FIG. 14 provides an X-ray diffraction pattern of Compound 1
Form II, 2-Methyltetrahydrofuran Solvate.
[0063] FIG. 15 is a differential scanning calorimetry (DSC) trace
of Compound 1 Form II, Acetone Solvate.
[0064] FIG. 16 is a Thermogravimetric analysis (TGA) plot of
Compound 1 Form II, Acetone Solvate.
[0065] FIG. 17 is a conformational image of Compound 1 Form II,
Acetone Solvate based on single crystal X-ray analysis.
[0066] FIG. 18 is a conformational image of the dimer of Compound 1
HCl Salt Form A.
[0067] FIG. 19 is an X-ray diffraction pattern of Compound 1 HCl
Salt Form A calculated from the crystal structure.
[0068] FIG. 20 is an .sup.1HNMR spectrum of Compound 1.
[0069] FIG. 21 is an .sup.1HNMR spectrum of Compound 1 HCl
salt.
[0070] FIG. 22 is a differential scanning calorimetry (DSC) trace
of Compound 1 Form I.
[0071] FIG. 23 is a conformational picture of Compound 1 Form I
based on single crystal X-ray analysis.
[0072] FIG. 24 is a conformational image of Compound 1 Form II,
Acetone Solvate, based on single crystal X-ray analysis.
[0073] FIG. 25 is a solid state .sup.13C NMR spectrum (15.0 kHz
spinning) of Compound 1 Form II, Acetone Solvate.
[0074] FIG. 26 is a solid state .sup.19F NMR spectrum (12.5 kHz
spinning) of Compound 1 Form II, Acetone Solvate.
[0075] FIG. 27 is an X-ray diffraction pattern of Compound 1 HCl
Salt Form A calculated from the crystal structure.
DETAILED DESCRIPTION
Definitions
[0076] As used herein, the term "active pharmaceutical ingredient"
or "API" refers to a biologically active compound. Exemplary APIs
include
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid (Compound 1).
[0077] The terms "solid form", "solid forms" and related terms,
when used herein to refer to
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl) benzoic acid (Compound 1), refer to a solid
form e.g. crystals and the like, comprising Compound 1 which is not
predominantly in a liquid or a gaseous state.
[0078] As used herein, the term "substantially amorphous" refers to
a solid material having little or no long range order in the
position of its molecules. For example, substantially amorphous
materials have less than about 15% crystallinity (e.g., less than
about 10% crystallinity or less than about 5% crystallinity). It is
also noted that the term `substantially amorphous` includes the
descriptor, `amorphous`, which refers to materials having no (0%)
crystallinity.
[0079] As used herein, the term "substantially crystalline" (as in
the phrase substantially crystalline Compound 1 Form I, Compound 1
Form II, or Compound 1 HCl Salt Form A) refers to a solid material
having predominantly long range order in the position of its
molecules. For example, substantially crystalline materials have
more than about 85% crystallinity (e.g., more than about 90%
crystallinity or more than about 95% crystallinity). It is also
noted that the term `substantially crystalline` includes the
descriptor, `crystalline`, which refers to materials having 100%
crystallinity.
[0080] The term "crystalline" and related terms used herein, when
used to describe a substance, component, product, or form, means
that the substance, component or product is substantially
crystalline as determined by X-ray diffraction. (See, e.g.,
Remington: The Science and Practice of Pharmacy, 21st Ed.,
Lippincott Williams & Wilkins, Baltimore, Md. (2003); The
United States Pharmacopeia, 23.sup.rd ed., 1843-1844 (1995)).
[0081] As used herein, the term "composition" generally refers to a
composition of two or more components, usually one or more drugs
(e.g., one drug (e.g., Compound 1 Form I, Compound 1 Form II, or
Compound 1 HCl Salt Form A)) and one or more pharmaceutical
excipients.
[0082] As used herein, the term "solid dosage form" generally
refers to a pharmaceutical composition, which when used in an oral
mode of administration include capsules, tablets, pills, powders
and granules. In such solid dosage forms, the active compound is
mixed with at least one inert, pharmaceutically acceptable
excipient or carrier.
[0083] As used herein, an "excipient" includes functional and
non-functional ingredients in a pharmaceutical composition.
[0084] As used herein, a "disintegrant" is an excipient that
hydrates a pharmaceutical composition and aids in tablet
dispersion. As used herein, a "diluent" or "filler" is an excipient
that adds bulkiness to a pharmaceutical composition.
[0085] As used herein, a "surfactant" is an excipient that imparts
pharmaceutical compositions with enhanced solubility and/or
wetability.
[0086] As used herein, a "binder" is an excipient that imparts a
pharmaceutical composition with enhanced cohesion or tensile
strength (e.g., hardness).
[0087] As used herein, a "glidant" is an excipient that imparts a
pharmaceutical compositions with enhanced flow properties.
[0088] As used herein, a "colorant" is an excipient that imparts a
pharmaceutical composition with a desired color. Examples of
colorants include commercially available pigments such as FD&C
Blue #1 Aluminum Lake, FD&C Blue #2, other FD&C Blue
colors, titanium dioxide, iron oxide, and/or combinations thereof.
In one embodiment, the pharmaceutical composition provided by the
invention is purple.
[0089] As used herein, a "lubricant" is an excipient that is added
to pharmaceutical compositions that are pressed into tablets. The
lubricant aids in compaction of granules into tablets and ejection
of a tablet of a pharmaceutical composition from a die press.
[0090] As used herein, "cubic centimeter" and "cc" are used
interchangeably to represent a unit of volume. Note that 1 cc=1
mL.
[0091] As used herein, "kiloPond" and "kP" are used interchangeably
and refer to the measure of force where a kP=approximately 9.8
Newtons.
[0092] As used herein, "friability" refers to the property of a
tablet to remain intact and withhold its form despite an external
force of pressure. Friability can be quantified using the
mathematical expression presented in equation 1:
% friabiliy = 100 .times. ( W 0 - W f ) W 0 ( 1 ) ##EQU00001##
wherein W.sub.0 is the original weight of the tablet and W.sub.f is
the final weight of the tablet after it is put through the
friabilator. Friability is measured using a standard USP testing
apparatus that tumbles experimental tablets for 100 or 400
revolutions. Some tablets of the invention have a friability of
less than 5.0%. In another embodiment, the friability is less than
2.0%. In another embodiment, the target friability is less than
1.0% after 400 revolutions.
[0093] As used herein, "mean particle diameter" is the average
particle diameter as measured using techniques such as laser light
scattering, image analysis, or sieve analysis. In one embodiment,
the granules used to prepare the pharmaceutical compositions
provided by the invention have a mean particle diameter of less
than 1.0 mm.
[0094] As used herein, "bulk density" is the mass of particles of
material divided by the total volume the particles occupy. The
total volume includes particle volume, inter-particle void volume
and internal pore volume. Bulk density is not an intrinsic property
of a material; it can change depending on how the material is
processed. In one embodiment, the granules used to prepare the
pharmaceutical compositions provided by the invention have a bulk
density of about 0.5-0.7 g/cc.
[0095] An effective amount or "therapeutically effective amount" of
a drug compound of the invention may vary according to factors such
as the disease state, age, and weight of the subject, and the
ability of the compound of the invention to elicit a desired
response in the subject. Dosage regimens may be adjusted to provide
the optimum therapeutic response. An effective amount is also one
in which any toxic or detrimental effects (e.g., side effects) of
the compound of the invention are outweighed by the therapeutically
beneficial effects.
[0096] As used herein, and unless otherwise specified, the terms
"therapeutically effective amount" and "effective amount" of a
compound mean an amount sufficient to provide a therapeutic benefit
in the treatment or management of a disease or disorder, or to
delay or minimize one or more symptoms associated with the disease
or disorder. A "therapeutically effective amount" and "effective
amount" of a compound mean an amount of therapeutic agent, alone or
in combination with one or more other agent(s), which provides a
therapeutic benefit in the treatment or management of the disease
or disorder. The terms "therapeutically effective amount" and
"effective amount" can encompass an amount that improves overall
therapy, reduces or avoids symptoms or causes of disease or
disorder, or enhances the therapeutic efficacy of another
therapeutic agent.
[0097] "Substantially pure" as used in the phrase "substantially
pure Compound 1 Form I, Compound 1 Form II, or Compound 1 HCl Salt
Form A," means greater than about 90% purity. In another
embodiment, substantially pure refers to greater than about 95%
purity. In another embodiment, substantially pure refers to greater
than about 98% purity. In another embodiment, substantially pure
refers to greater than about 99% purity.
[0098] With respect to Compound 1 (e.g., Compound 1 Form I,
Compound 1 Form II, Compound 1 HCl Salt Form A), the terms "about"
and "approximately", when used in connection with doses, amounts,
or weight percent of ingredients of a composition or a dosage form,
mean a dose, amount, or weight percent that is recognized by one of
ordinary skill in the art to provide a pharmacological effect
equivalent to that obtained from the specified dose, amount, or
weight percent. Specifically the term "about" or "approximately"
means an acceptable error for a particular value as determined by
one of ordinary skill in the art, which depends in part on how the
value is measured or determined. In certain embodiments, the term
"about" or "approximately" means within 1, 2, 3, or 4 standard
deviations. In certain embodiments, the term "about" or
"approximately" means within 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%,
6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, or 0.05% of a given value or
range.
[0099] Unless otherwise specified, the term "Compound 1" includes,
but is not limited to, the solid forms of Compound 1 as described
herein, e.g. Compound 1 Form I, Compound 1 Form II, or Compound 1
HCl Salt Form A, as well as combinations thereof.
Pharmaceutical Compositions
[0100] The invention provides pharmaceutical compositions,
pharmaceutical formulations and solid dosage forms comprising
Compound 1 which may be in substantially crystalline form. In some
embodiments, Compound 1 is in crystalline Form I (Compound 1 Form
I). In some embodiments, Compound 1 is in crystalline Form II
(Compound 1 Form II). In some embodiments, Compound 1 is in
crystalline HCl salt form (Compound 1 HCl Salt Form A). In some
embodiments of this aspect, the amount of Compound 1 that is
present in the pharmaceutical composition is 25 mg, 50 mg, 75 mg,
100 mg, 125 mg, 150 mg, 200 mg, 250 mg, or 400 mg. In some
embodiments of this aspect, weight/weight relative percent of
Compound 1 that is present in the pharmaceutical composition is
from 10 to 75 percent. In these and other embodiments,
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid is present as substantially pure
Compound 1. "Substantially pure" means greater than ninety percent
pure; preferably greater than 95 percent pure; more preferably
greater than 99.5 percent pure (i.e., not mixed with other
crystalline forms of Compound 1).
[0101] Thus in one aspect, the invention provides a pharmaceutical
composition comprising:
[0102] a. Compound 1;
[0103] b. a filler;
[0104] c. a disintegrant;
[0105] d. a surfactant;
[0106] e. a diluent;
[0107] f. a lubricant; and
[0108] g. and at least one of a glidant and a binder.
[0109] In one embodiment of this aspect, the pharmaceutical
composition comprises 25 mg of Compound 1. In another embodiment of
this aspect, the pharmaceutical composition comprises 50 mg of
Compound 1. In another embodiment of this aspect, the
pharmaceutical composition comprises 100 mg of Compound 1. In
another embodiment of this aspect, the pharmaceutical composition
comprises 125 mg of Compound 1. In another embodiment of this
aspect, the pharmaceutical composition comprises 150 mg of Compound
1. In another embodiment of this aspect, the pharmaceutical
composition comprises 200 mg of Compound 1. In another embodiment
of this aspect, the pharmaceutical composition comprises 250 mg of
Compound 1. In another embodiment of this aspect, the
pharmaceutical composition comprises 400 mg of Compound 1.
[0110] In some embodiments, the pharmaceutical compositions
comprises Compound 1, wherein Compound 1 is present in an amount of
at least 15 wt % (e.g., at least 20 wt %, at least 30 wt %, at
least 40 wt %, at least 50 wt %, or at least 60 wt %) by weight of
the composition.
[0111] In some embodiments, the pharmaceutical composition
comprises Compound 1, a filler, a diluent, a disintegrant, a
surfactant, a glidant, and a lubricant. In this embodiment, the
composition comprises from about 20 wt % to about 50 wt % (e.g.,
about 25-35 wt %) of Compound 1 by weight of the composition, and
more typically, from 25 wt % to about 45 wt % (e.g., about 28-32 wt
%) of Compound 1 by weight of the composition.
[0112] In some embodiments, the pharmaceutical composition
comprises Compound 1, a filler, a diluent, a disintegrant, a
surfactant, a binder, and a lubricant. In this embodiment, the
composition comprises from about 30 wt % to about 60 wt % (e.g.,
about 40-55 wt %) of Compound 1 by weight of the composition, and
more typically from 35 wt % to about 70 wt % (e.g., about 45-55 wt
%) of Compound 1 by weight of the composition.
[0113] The concentration of Compound 1 in the composition depends
on several factors such as the amount of pharmaceutical composition
needed to provide a desired amount of Compound 1 and the desired
dissolution profile of the pharmaceutical composition.
[0114] In another embodiment, the pharmaceutical composition
comprises Compound 1, in which Compound 1 in its solid form has a
mean particle diameter, measured by light scattering (e.g., using a
Malvern Mastersizer available from Malvern Instruments in England)
of 0.1 microns to 10 microns. In another embodiment, the particle
size of Compound 1 is 1 micron to 5 microns. In another embodiment,
Compound 1 has a particle size D50 of 2.0 microns.
[0115] As indicated, in addition to Compound 1, in some embodiments
of the invention, the pharmaceutical compositions which are oral
formulations also comprise one or more excipients such as fillers,
disintegrants, surfactants, diluents, binders, glidants,
lubricants, colorants, or fragrances and any combination
thereof.
[0116] Fillers suitable for the invention are compatible with the
ingredients of the pharmaceutical composition, i.e., they do not
substantially reduce the solubility, the hardness, the chemical
stability, the physical stability, or the biological activity of
the pharmaceutical composition. Exemplary fillers include:
celluloses, modified celluloses, (e.g. sodium carboxymethyl
cellulose, ethyl cellulose hydroxymethyl cellulose,
hydroxypropylcellulose), cellulose acetate, microcrystalline
cellulose, calcium phosphates, dibasic calcium phosphate, starches
(e.g. corn starch, potato starch), sugars (e.g., sorbitol) lactose,
sucrose, or the like), or any combination thereof.
[0117] Thus, in one embodiment, the pharmaceutical composition
comprises at least one filler in an amount of at least 5 wt %
(e.g., at least about 20 wt %, at least about 30 wt %, or at least
about 40 wt %) by weight of the composition. For example, the
pharmaceutical composition comprises from about 10 wt % to about 60
wt % (e.g., from about 20 wt % to about 55 wt %, from about 25 wt %
to about 50 wt %, or from about 27 wt % to about 45 wt %) of
filler, by weight of the composition. In another example, the
pharmaceutical composition comprises at least about 20 wt % (e.g.,
at least 30 wt % or at least 40 wt %) of microcrystalline
cellulose, for example MCC Avicel PH102, by weight of the
composition. In yet another example, the pharmaceutical composition
comprises from about 10 wt % to about 60 wt % (e.g., from about 20
wt % to about 55 wt % or from about 25 wt % to about 45 wt %) of
microcellulose, by weight of the composition.
[0118] Disintegrants suitable for the invention enhance the
dispersal of the pharmaceutical composition and are compatible with
the ingredients of the pharmaceutical composition, i.e., they do
not substantially reduce the chemical stability, the physical
stability, the hardness, or the biological activity of the
pharmaceutical composition. Exemplary disintegrants include
croscarmellose sodium, sodium starch glycolate, or a combination
thereof.
[0119] Thus, in one embodiment, the pharmaceutical composition
comprises disintegrant in an amount of about 10 wt % or less (e.g.,
about 7 wt % or less, about 6 wt % or less, or about 5 wt % or
less) by weight of the composition. For example, the pharmaceutical
composition comprises from about 1 wt % to about 10 wt % (e.g.,
from about 1.5 wt % to about 7.5 wt % or from about 2.5 wt % to
about 6 wt %) of disintegrant, by weight of the composition. In
another example, the pharmaceutical composition comprises about 10
wt % or less (e.g., 7 wt % or less, 6 wt % or less, or 5 wt % or
less) of croscarmellose sodium, by weight of the composition. In
yet another example, the pharmaceutical composition comprises from
about 1 wt % to about 10 wt % (e.g., from about 1.5 wt % to about
7.5 wt % or from about 2.5 wt % to about 6 wt %) of croscarmellose
sodium, by weight of the composition. In some examples, the
pharmaceutical composition comprises from about 0.1% to about 10 wt
% (e.g., from about 0.5 wt % to about 7.5 wt % or from about 1.5 wt
% to about 6 wt %) of disintegrant, by weight of the composition.
In still other examples, the pharmaceutical composition comprises
from about 0.5% to about 10 wt % (e.g., from about 1.5 wt % to
about 7.5 wt % or from about 2.5 wt % to about 6 wt %) of
disintegrant, by weight of the composition.
[0120] Surfactants suitable for the invention enhance the
wettability of the pharmaceutical composition and are compatible
with the ingredients of the pharmaceutical composition, i.e., they
do not substantially reduce the chemical stability, the physical
stability, the hardness, or the biological activity of the
pharmaceutical composition. Exemplary surfactants include sodium
lauryl sulfate (SLS), sodium stearyl fumarate (SSF),
polyoxyethylene 20 sorbitan mono-oleate (e.g., Tween.TM.), any
combination thereof, or the like.
[0121] Thus, in one embodiment, the pharmaceutical composition
comprises a surfactant in an amount of about 10 wt % or less (e.g.,
about 5 wt % or less, about 2 wt % or less, about 1 wt % or less,
about 0.8 wt % or less, or about 0.6 wt % or less) by weight of the
composition. For example, the pharmaceutical composition includes
from about 10 wt % to about 0.1 wt % (e.g., from about 5 wt % to
about 0.2 wt % or from about 2 wt % to about 0.3 wt %) of
surfactant, by weight of the composition. In another example, the
pharmaceutical composition comprises 10 wt % or less (e.g., about 5
wt % or less, about 2 wt % or less, about 1 wt % or less, about 0.8
wt % or less, or about 0.6 wt % or less) of sodium lauryl sulfate,
by weight of the composition. In yet another example, the
pharmaceutical composition comprises from about 10 wt % to about
0.1 wt % (e.g., from about 5 wt % to about 0.2 wt % or from about 2
wt % to about 0.3 wt %) of sodium lauryl sulfate, by weight of the
composition.
[0122] Binders suitable for the invention enhance the tablet
strength of the pharmaceutical composition and are compatible with
the ingredients of the pharmaceutical composition, i.e., they do
not substantially reduce the chemical stability, the physical
stability, or the biological activity of the pharmaceutical
composition. Exemplary binders include polyvinylpyrrolidone,
dibasic calcium phosphate, sucrose, corn (maize) starch, modified
cellulose (e.g., hydroxymethyl cellulose), or any combination
thereof.
[0123] Thus, in one embodiment, the pharmaceutical composition
comprises a binder in an amount of at least about 0.1 wt % (e.g.,
at least about 1 wt %, at least about 3 wt %, at least about 4 wt
%, or at least about 5 wt %) by weight of the composition. For
example, the pharmaceutical composition comprises from about 0.1 wt
% to about 10 wt % (e.g., from about 1 wt % to about 10 wt % or
from about 2 wt % to about 7 wt %) of binder, by weight of the
composition. In another example, the pharmaceutical composition
comprises at least about 0.1 wt % (e.g., at least about 1 wt %, at
least about 2 wt %, at least about 3 wt %, or at least about 4 wt
%) of polyvinylpyrrolidone, by weight of the composition. In yet
another example, the pharmaceutical composition comprises a glidant
in an amount ranging from about 0.1 wt % to about 10 wt % (e.g.,
from about 1 wt % to about 8 wt % or from about 2 wt % to about 5
wt %) of polyvinylpyrrolidone, by weight of the composition.
[0124] Diluents suitable for the invention may add necessary bulk
to a formulation to prepare tablets of the desired size and are
generally compatible with the ingredients of the pharmaceutical
composition, i.e., they do not substantially reduce the solubility,
the hardness, the chemical stability, the physical stability, or
the biological activity of the pharmaceutical composition.
Exemplary diluents include: sugars, for example, confectioner's
sugar, compressible sugar, dextrates, dextrin, dextrose, lactose,
mannitol, sorbitol, cellulose, and modified celluloses, for
example, powdered cellulose, talc, calcium phosphate, starch, or
any combination thereof.
[0125] Thus, in one embodiment, the pharmaceutical composition
comprises a diluent in an amount of 40 wt % or less (e.g., 35 wt %
or less, 30 wt % or less, or 25 wt % or less, or 20 wt % or less,
or 15 wt % or less, or 10 wt % or less) by weight of the
composition. For example, the pharmaceutical composition comprises
from about 40 wt % to about 1 wt % (e.g., from about 35 wt % to
about 5 wt % or from about 30 wt % to about 7 wt %, from about 25
wt % to about 10 wt %, from about 20 wt % to about 15 wt %) of
diluent, by weight of the composition. In another example, the
pharmaceutical composition comprises 40 wt % or less (e.g., 35 wt %
or less, 25 wt % or less, or 15 wt % or less) of mannitol, by
weight of the composition. In yet another example, the
pharmaceutical composition comprises from about 35 wt % to about 1
wt % (e.g., from about 30 wt % to about 5 wt % or from about 25 wt
% to about 10 wt %) of mannitol, by weight of the composition.
[0126] Glidants suitable for the invention enhance the flow
properties of the pharmaceutical composition and are compatible
with the ingredients of the pharmaceutical composition, i.e., they
do not substantially reduce the solubility, the hardness, the
chemical stability, the physical stability, or the biological
activity of the pharmaceutical composition. Exemplary glidants
include colloidal silicon dioxide, talc, or a combination
thereof.
[0127] Thus, in one embodiment, the pharmaceutical composition
comprises a glidant in an amount of 2 wt % or less (e.g., 1.75 wt
%, 1.25 wt % or less, or 1.00 wt % or less) by weight of the
composition. For example, the pharmaceutical composition comprises
from about 2 wt % to about 0.05 wt % (e.g., from about 1.5 wt % to
about 0.07 wt % or from about 1.0 wt % to about 0.09 wt %) of
glidant, by weight of the composition. In another example, the
pharmaceutical composition comprises 2 wt % or less (e.g., 1.75 wt
%, 1.25 wt % or less, or 1.00 wt % or less) of colloidal silicon
dioxide, by weight of the composition. In yet another example, the
pharmaceutical composition comprises from about 2 wt % to about
0.05 wt % (e.g., from about 1.5 wt % to about 0.07 wt % or from
about 1.0 wt % to about 0.09 wt %) of colloidal silicon dioxide, by
weight of the composition.
[0128] In some embodiments, the pharmaceutical composition can
include an oral solid pharmaceutical dosage form which can comprise
a lubricant that can prevent adhesion of a granulate-bead admixture
to a surface (e.g., a surface of a mixing bowl, a compression die
and/or punch). A lubricant can also reduce interparticle friction
within the granulate and improve the compression and ejection of
compressed pharmaceutical compositions from a die press. The
lubricant is also compatible with the ingredients of the
pharmaceutical composition, i.e., they do not substantially reduce
the solubility, the hardness, or the biological activity of the
pharmaceutical composition. Exemplary lubricants include magnesium
stearate, calcium stearate, zinc stearate, sodium stearate, stearic
acid, aluminum stearate, leucine, glyceryl behenate, hydrogenated
vegetable oil or any combination thereof. In one embodiment, the
pharmaceutical composition comprises a lubricant in an amount of 5
wt % or less (e.g., 4.75 wt %, 4.0 wt % or less, or 3.00 wt % or
less, or 2.0 wt % or less) by weight of the composition. For
example, the pharmaceutical composition comprises from about 5 wt %
to about 0.10 wt % (e.g., from about 4.5 wt % to about 0.5 wt % or
from about 3 wt % to about 1 wt %) of lubricant, by weight of the
composition. In another example, the pharmaceutical composition
comprises 5 wt % or less (e.g., 4.0 wt % or less, 3.0 wt % or less,
or 2.0 wt % or less, or 1.0 wt % or less) of magnesium stearate, by
weight of the composition. In yet another example, the
pharmaceutical composition comprises from about 5 wt % to about
0.10 wt % (e.g., from about 4.5 wt % to about 0.15 wt % or from
about 3.0 wt % to about 0.50 wt %) of magnesium stearate, by weight
of the composition.
[0129] Pharmaceutical compositions of the invention can optionally
comprise one or more colorants, flavors, and/or fragrances to
enhance the visual appeal, taste, and/or scent of the composition.
Suitable colorants, flavors, or fragrances are compatible with the
ingredients of the pharmaceutical composition, i.e., they do not
substantially reduce the solubility, the chemical stability, the
physical stability, the hardness, or the biological activity of the
pharmaceutical composition. In one embodiment, the pharmaceutical
composition comprises a colorant, a flavor, and/or a fragrance. In
one embodiment, the pharmaceutical compositions provided by the
invention are purple.
[0130] In some embodiments, the pharmaceutical composition includes
or can be made into tablets and the tablets can be coated with a
colorant and optionally labeled with a logo, other image and/or
text using a suitable ink. In still other embodiments, the
pharmaceutical composition includes or can be made into tablets and
the tablets can be coated with a colorant, waxed, and optionally
labeled with a logo, other image and/or text using a suitable ink.
Suitable colorants and inks are compatible with the ingredients of
the pharmaceutical composition, i.e., they do not substantially
reduce the solubility, the chemical stability, the physical
stability, the hardness, or the biological activity of the
pharmaceutical composition. The suitable colorants and inks can be
any color and are water based or solvent based. In one embodiment,
tablets made from the pharmaceutical composition are coated with a
colorant and then labeled with a logo, other image, and/or text
using a suitable ink. For example, tablets comprising
pharmaceutical composition as described herein can be coated with
about 3 wt % (e.g., less than about 6 wt % or less than about 4 wt
%) of film coating comprising a colorant. The colored tablets can
be labeled with a logo and text indicating the strength of the
active ingredient in the tablet using a suitable ink. In another
example, tablets comprising pharmaceutical composition as described
herein can be coated with about 3 wt % (e.g., less than about 6 wt
% or less than about 4 wt %) of a film coating comprising a
colorant.
[0131] In another embodiment, tablets made from the pharmaceutical
composition are coated with a colorant, waxed, and then labeled
with a logo, other image, and/or text using a suitable ink. For
example, tablets comprising pharmaceutical composition as described
herein can be coated with about 3 wt % (e.g., less than about 6 wt
% or less than about 4 wt %) of film coating comprising a colorant.
The colored tablets can be waxed with Carnauba wax powder weighed
out in the amount of about 0.01% w/w of the starting tablet core
weight. The waxed tablets can be labeled with a logo and text
indicating the strength of the active ingredient in the tablet
using a suitable ink. In another example, tablets comprising
pharmaceutical composition as described herein can be coated with
about 3 wt % (e.g., less than about 6 wt % or less than about 4 wt
%) of a film coating comprising a colorant The colored tablets can
be waxed with Carnauba wax powder weighed out in the amount of
about 0.01% w/w of the starting tablet core weight. The waxed
tablets can be labeled with a logo and text indicating the strength
of the active ingredient in the tablet using a pharmaceutical grade
ink such as a black ink (e.g., Opacode.RTM. S-1-17823, a solvent
based ink, commercially available from Colorcon, Inc. of West
Point, Pa.).
[0132] One exemplary pharmaceutical composition comprises from
about 15 wt % to about 70 wt % (e.g., from about 15 wt % to about
60 wt %, from about 15 wt % to about 50 wt %, or from about 15 wt %
to about 40 wt %, or from about 20 wt % to about 70 wt %, or from
about 30 wt % to about 70 wt %, or from about 40 wt % to about 70
wt %, or from about 50 wt % to about 70 wt %) of Compound 1, by
weight of the composition. The aforementioned compositions can also
include one or more pharmaceutically acceptable excipients, for
example, from about 20 wt % to about 50 wt % of a filler; from
about 1 wt % to about 5 wt % of a disintegrant; from about 2 wt %
to about 0.3 wt % of a surfactant; from about 0.1 wt % to about 5
wt % of a binder; from about 1 wt % to about 30 wt % of a diluent;
from about 2 wt % to about 0.05 wt % of a glidant; and from about 5
wt % to about 0.1 wt % of a lubricant. Or, the pharmaceutical
composition comprises a composition containing from about 15 wt %
to about 70 wt % (e.g., from about 20 wt % to about 40 wt %, from
about 25 wt % to about 60 wt %, or from about 30 wt % to about 55
wt %) of Compound 1, by weight of the composition; and one or more
excipients, for example, from about 20 wt % to about 50 wt % of a
filler; from about 1 wt % to about 5 wt % of a disintegrant; from
about 2 wt % to about 0.3 wt % of a surfactant; from about 0.1 wt %
to about 5 wt % of a binder; from about 1 wt % to about 30 wt % of
a diluent; from about 2 wt % to about 0.05 wt % of a glidant; and
from about 5 wt % to about 0.1 wt % of a lubricant.
[0133] Another exemplary pharmaceutical composition comprises from
about 15 wt % to about 70 wt % (e.g., from about 15 wt % to about
60 wt %, from about 15 wt % to about 50 wt %, or from about 15 wt %
to about 40 wt % or from about 20 wt % to about 70 wt %, or from
about 30 wt % to about 70 wt %, or from about 40 wt % to about 70
wt %, or from about 50 wt % to about 70 wt %) of Compound 1 by
weight of the composition, and one or more excipients, for example,
from about 20 wt % to about 50 wt % of a filler; from about 1 wt %
to about 5 wt % of a disintegrant; from about 2 wt % to about 0.3
wt % of a surfactant; from about 0.1 wt % to about 5 wt % of a
binder; from about 1 wt % to about 30 wt % of a diluent; from about
2 wt % to about 0.05 wt % of a glidant; and from about 2 wt % to
about 0.1 wt % of a lubricant.
[0134] Another exemplary pharmaceutical composition comprises from
about 15 wt % to about 70 wt % (e.g., from about 15 wt % to about
60 wt %, from about 15 wt % to about 50 wt %, or from about 15 wt %
to about 40 wt % or from about 20 wt % to about 70 wt %, or from
about 30 wt % to about 70 wt %, or from about 40 wt % to about 70
wt %, or from about 50 wt % to about 70 wt %) of Compound 1 by
weight of the composition, and one or more excipients, for example,
from about 20 wt % to about 50 wt % of a filler; from about 1 wt %
to about 5 wt % of a disintegrant; from about 2 wt % to about 0.3
wt % of a surfactant; from about 0.1 wt % to about 5 wt % of a
binder; from about 1 wt % to about 30 wt % of a diluent; from about
2 wt % to about 0.05 wt % of a glidant; and from about 2 wt % to
about 0.1 wt % of a lubricant.
[0135] Another exemplary pharmaceutical composition comprises from
about 15 wt % to about 70 wt % (e.g., from about 15 wt % to about
60 wt %, from about 15 wt % to about 50 wt %, or from about 15 wt %
to about 40 wt % or from about 20 wt % to about 70 wt %, or from
about 30 wt % to about 70 wt %, or from about 40 wt % to about 70
wt %, or from about 50 wt % to about 70 wt %) of Compound 1 and one
or more excipients, for example, from about 20 wt % to about 50 wt
% of a filler; from about 1 wt % to about 5 wt % of a disintegrant;
from about 2 wt % to about 0.3 wt % of a surfactant; from about 0.1
wt % to about 5 wt % of a binder; from about 1 wt % to about 30 wt
% of a diluent; from about 2 wt % to about 0.05 wt % of a glidant;
and from about 2 wt % to about 0.1 wt % of a lubricant.
[0136] In one embodiment, the invention is a granular
pharmaceutical composition comprising:
[0137] a. about 30 wt % of Compound 1 by weight of the
composition;
[0138] b. about 42 wt % of microcrystalline cellulose by weight of
the composition;
[0139] c. about 21 wt % of mannitol by weight of the
composition;
[0140] d. about 3 wt % of sodium croscarmellose sodium by weight of
the composition;
[0141] e. about 1 wt % of sodium lauryl sulfate by weight of the
composition;
[0142] f. about 2 wt % of magnesium stearate by weight of the
composition; and
[0143] g. about 0.5 wt % of colloidal silica by weight of the
composition.
[0144] Another granular composition formulated into an oral
formulation of the invention comprises:
[0145] a. about 50 wt % of Compound 1;
[0146] b. about 30 wt % of microcrystalline cellulose by weight of
the composition;
[0147] c. about 13 wt % of mannitol by weight of the
composition;
[0148] d. about 2 wt % of sodium croscarmellose sodium by weight of
the composition;
[0149] e. about 4 wt % of polyvinylpyrrolidone by weight of the
composition; and
[0150] f. about 1 wt % of sodium lauryl sulfate by weight of the
composition.
[0151] In one embodiment, a pharmaceutical oral formulation of the
invention comprises:
[0152] a. about 30 wt % of a Compound 1 by weight of the
composition;
[0153] b. about 42 wt % of microcrystalline cellulose by weight of
the composition;
[0154] c. about 21 wt % of mannitol by weight of the
composition;
[0155] d. about 3 wt % of sodium croscarmellose sodium by weight of
the composition;
[0156] e. about 1 wt % of sodium lauryl sulfate by weight of the
composition;
[0157] f. about 2.5 wt % of magnesium stearate by weight of the
composition; and
[0158] g. about 0.5 wt % of colloidal silica by weight of the
composition.
[0159] Another pharmaceutical oral formulation of the invention
comprises:
[0160] a. about 50 wt % of a Compound 1 by weight of the
composition;
[0161] b. about 30 wt % of microcrystalline cellulose by weight of
the composition;
[0162] c. about 13 wt % of mannitol by weight of the
composition;
[0163] d. about 4 wt % of sodium croscarmellose sodium by weight of
the composition;
[0164] e. about 4 wt % of polyvinylpyrrolidone by weight of the
composition
[0165] f. about 1 wt % of sodium lauryl sulfate by weight of the
composition; and
[0166] g. about 0.5 wt % of magnesium stearate by weight of the
composition.
[0167] Another pharmaceutical oral formulation of the invention
comprises:
[0168] a. about 60 wt % of a Compound 1 by weight of the
composition;
[0169] b. about 20 wt % of microcrystalline cellulose by weight of
the composition;
[0170] c. about 13 wt % of mannitol by weight of the
composition;
[0171] d. about 4 wt % of sodium croscarmellose sodium by weight of
the composition;
[0172] e. about 4 wt % of polyvinylpyrrolidone by weight of the
composition
[0173] f. about 1 wt % of sodium lauryl sulfate by weight of the
composition; and
[0174] g. about 0.5 wt % of magnesium stearate by weight of the
composition.
[0175] Another pharmaceutical oral formulation of the invention
comprises:
[0176] a. about 150 to 250 mg of Compound 1;
[0177] b. about 40 to 50 mg of mannitol;
[0178] c. about 120 to 130 mg of microcrystalline cellulose;
[0179] d. about 10 to 20 mg of croscarmellose sodium;
[0180] e. about 10 to 20 mg of polyvinylpyrrolidone;
[0181] f. about 1 to 5 mg of sodium lauryl sulfate; and
[0182] g. about 1 to 5 mg of magnesium stearate.
[0183] Another pharmaceutical oral formulation of the invention
comprises:
[0184] a. about 200 mg of Compound 1;
[0185] b. about 43 mg of mannitol;
[0186] c. about 123 mg of microcrystalline cellulose;
[0187] d. about 15 mg of croscarmellose sodium;
[0188] e. about 13 mg of polyvinylpyrrolidone;
[0189] f. about 3 mg of sodium lauryl sulfate; and
[0190] g. about 4 mg of magnesium stearate.
[0191] Another pharmaceutical oral formulation of the invention
comprises:
[0192] a. about 200 mg of Compound 1;
[0193] b. about 45 mg of mannitol;
[0194] c. about 123 mg of microcrystalline cellulose;
[0195] d. about 15 mg of croscarmellose sodium;
[0196] e. about 10.4 mg of polyvinylpyrrolidone;
[0197] f. about 2.6 mg of sodium lauryl sulfate; and
[0198] g. about 4 mg of magnesium stearate.
[0199] Another pharmaceutical oral formulation of the invention
comprises:
[0200] a. about 70 wt % of a Compound 1 by weight of the
composition;
[0201] b. about 12 wt % of microcrystalline cellulose by weight of
the composition;
[0202] c. about 11 wt % of mannitol by weight of the
composition;
[0203] d. about 4 wt % of sodium croscarmellose sodium by weight of
the composition;
[0204] e. about 4 wt % of polyvinylpyrrolidone by weight of the
composition
[0205] f. about 1 wt % of sodium lauryl sulfate by weight of the
composition; and
[0206] g. about 0.5 wt % of magnesium stearate by weight of the
composition.
[0207] The pharmaceutical compositions of the invention can be
processed into a tablet form, capsule form, pouch form, lozenge
form, or other solid form that is suited for oral administration.
Thus in some embodiments, the pharmaceutical compositions are in
tablet form.
[0208] In still another pharmaceutical oral formulation of the
invention, a shaped pharmaceutical tablet composition having an
initial hardness of 5-21 kP.+-.20 percent comprises: about 30 wt %
of Compound 1; about 42 wt % of microcrystalline cellulose by
weight of the composition; about 21 wt % of mannitol by weight of
the composition; about 3 wt % of sodium croscarmellose sodium by
weight of the composition; about 1 wt % of sodium lauryl sulfate by
weight of the composition; about 2.5 wt % of magnesium stearate by
weight of the composition; and about 0.5 wt % of colloidal silica
by weight of the composition. Wherein the amount of Compound 1 in
the shaped pharmaceutical tablet ranges from about 25 mg to about
250 mg, for example, 50 mg, or 75 mg, or 100 mg, or 150 mg, 200 mg,
or 250 mg Compound 1 per tablet.
[0209] In still another pharmaceutical oral formulation of the
invention, a shaped pharmaceutical tablet composition having an
initial hardness of 5-21 kP.+-.20 percent comprises: about 49 wt %
of a Compound 1; about 29 wt % of microcrystalline cellulose by
weight of the composition; about 12.6 wt % of mannitol by weight of
the composition; about 4 wt % of sodium croscarmellose sodium by
weight of the composition; about 4 wt % of polyvinylpyrrolidone by
weight of the composition; about 1 wt % of sodium lauryl sulfate by
weight of the composition; and about 0.5 wt % of magnesium stearate
by weight of the composition. The amount of Compound 1 in the
shaped pharmaceutical tablet ranges from about 25 mg to about 250
mg, for example, 50 mg, or 75 mg, or 100 mg, or 150 mg, 200 mg, or
250 mg Compound 1 per tablet.
[0210] In certain embodiments, the shaped pharmaceutical tablet
contains about 100 mg of Compound 1. In certain embodiments, the
shaped pharmaceutical tablet contains about 200 mg of Compound
1.
[0211] Another aspect of the invention provides a pharmaceutical
formulation consisting of a tablet or capsule that includes a
Compound 1 and other excipients (e.g., a filler, a disintegrant, a
surfactant, a binder, a glidant, a colorant, a lubricant, or any
combination thereof), each of which is described above and in the
Examples below, wherein the tablet has a dissolution of at least
about 50% (e.g., at least about 60%, at least about 70%, at least
about 80%, at least about 90%, or at least about 99%) in about 30
minutes. In one example, the pharmaceutical composition consists of
a tablet that includes Compound 1 in an amount ranging from 25 mg
to 250 mg, for example, 25 mg, or 50 mg, or 75 mg, or 100 mg, or
150 mg, 200 mg, or 250 mg and one or more excipients (e.g., a
filler, a disintegrant, a surfactant, a binder, a glidant, a
colorant, a lubricant, or any combination thereof), each of which
is described above and in the Examples below, wherein the tablet
has a dissolution of from about 50% to about 100% (e.g., from about
55% to about 95% or from about 60% to about 90%) in about 30
minutes. In another example, the pharmaceutical composition
consists of a tablet that comprises a composition comprising
Compound 1; and one or more excipients from: a filler, a diluent, a
disintegrant, a surfactant, a binder, a glidant, and a lubricant,
wherein the tablet has a dissolution of at least about 50% (e.g.,
at least about 60%, at least about 70%, at least about 80%, at
least about 90%, or at least about 99%) in about 30 minutes.
[0212] In one embodiment, the tablet comprises a composition
comprising at least about 25 mg (e.g., at least about 30 mg, at
least about 40 mg, or at least about 50 mg) of Compound 1; and one
or more excipients from: a filler, a diluent, a disintegrant, a
surfactant, a binder, a glidant, and a lubricant. In another
embodiment, the tablet comprises a composition comprising at least
about 25 mg (e.g., at least about 30 mg, at least about 40 mg, at
least about 50 mg, at least about 100 mg, or at least 150 mg) of
Compound 1 and one or more excipients from: a filler, a diluent, a
disintegrant, a surfactant, a binder, a glidant, and a
lubricant.
[0213] Dissolution can be measured with a standard USP Type II
apparatus that employs a dissolution media of 0.1% CTAB dissolved
in 900 mL of DI water, buffered at pH 6.8 with 50 mM potassium
phosphate monoasic, stirring at about 50-75 rpm at a temperature of
about 37.degree. C. A single experimental tablet is tested in each
test vessel of the apparatus. Dissolution can also be measured with
a standard USP Type II apparatus that employs a dissolution media
of 0.7% sodium lauryl sulfate dissolved in 900 mL of 50 mM sodium
phosphate buffer (pH 6.8), stirring at about 65 rpm at a
temperature of about 37.degree. C. A single experimental tablet is
tested in each test vessel of the apparatus. Dissolution can also
be measured with a standard USP Type II apparatus that employs a
dissolution media of 0.5% sodium lauryl sulfate dissolved in 900 mL
of 50 mM sodium phosphate buffer (pH 6.8), stirring at about 65 rpm
at a temperature of about 37.degree. C. A single experimental
tablet is tested in each test vessel of the apparatus.
Methods for Making Compound 1, Compound 1 Form I, Compound 1 Form
II, Compound 1 HCl Salt Form A
Compound 1
[0214] Compound 1 is used as the starting point for the other solid
state forms and can be prepared by coupling an acid chloride moiety
with an amine moiety according to Schemes 1-4.
##STR00002##
[0215] Scheme 1 depicts the preparation of
1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarbonyl
chloride, which is used in Scheme 3 to make the amide linkage of
Compound 1.
[0216] The starting material,
2,2-difluorobenzo[d][1,3]dioxole-5-carboxylic acid, is commercially
available from Saltigo (an affiliate of the Lanxess Corporation).
Reduction of the carboxylc acid moiety in
2,2-difluorobenzo[d][1,3]dioxole-5-carboxylic acid to the primary
alcohol, followed by conversion to the corresponding chloride using
thionyl chloride (SOCl.sub.2), provides
5-(chloromethyl)-2,2-difluorobenzo[d][1,3]dioxole, which is
subsequently converted to
2-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)acetonitrile using sodium
cyanide. Treatment of
2-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)acetonitrile with base and
1-bromo-2-chloroethane provides
1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarbonitrile.
The nitrile moiety in
1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarbonitrile is
converted to a carboxylic acid using base to give
1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxylic
acid, which is converted to the desired acid chloride using thionyl
chloride.
##STR00003##
[0217] Scheme 2 depicts an alternative synthesis of the requisite
acid chloride. 5-bromomethyl-2,2-difluoro-1,3-benzodioxole is
coupled with ethyl cyanoacetate in the presence of a palladium
catalyst to form the corresponding alpha cyano ethyl ester.
Saponification of the ester moiety to the carboxylic acid gives the
cyanoethyl compound. Alkylation of the cyanoethyl compound with
1-bromo-2-chloro ethane in the presence of base gives the
cyanocyclopropyl compound. Treatment of the cyanocyclopropyl
compound with base gives the carboxylate salt, which is converted
to the carboxylic acid by treatment with acid. Conversion of the
carboxylic acid to the acid chloride is then accomplished using a
chlorinating agent such as thionyl chloride or the like.
##STR00004##
[0218] Scheme 3 depicts the preparation of the requisite tert-butyl
3-(6-amino-3-methylpyridin-2-yl)benzoate, which is coupled with
1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarbonyl
chloride in Scheme 3 to give Compound 1. Palladium-catalyzed
coupling of 2-bromo-3-methylpyridine with
3-(tert-butoxycarbonyl)phenylboronic acid gives tert-butyl
3-(3-methylpyridin-2-yl)benzoate, which is subsequently converted
to the desired compound.
##STR00005##
[0219] Scheme 4 depicts the coupling of
1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarbonyl
chloride with tert-butyl 3-(6-amino-3-methylpyridin-2-yl)benzoate
using triethyl amine and 4-dimethylaminopyridine to initially
provide the tert-butyl ester of Compound 1.
Compound 1 Form I
[0220] Compound 1 Form I is prepared by dispersing or dissolving a
salt form, such as the HCl salt, of Compound 1 in an appropriate
solvent for an effective amount of time. Treatment of the
tert-butyl ester with an acid such as HCl, gives the HCL salt of
Compound 1, which is typically a crystalline solid. Compound 1 Form
I may also be prepared directly from the t-butyl ester precursor by
treatment with an appropriate acid, such as formic acid.
[0221] The HCl salt of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid can be used to make Form I by
dispersing or dissolving the HCl salt of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid in an appropriate solvent for an
effective amount of time. Other salts of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid may be used, such as, for example,
salts derived from other mineral or organic acids. The other salts
result from acid-mediated hydrolysis of the t-butyl ester moiety.
Salts derived from other acids may include, for example, nitric,
sulfuric, phosphoric, boric, acetic, benzoic and malonic. These
salt forms of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid may or may not be soluble,
depending upon the solvent used, but lack of solubility does not
hinder formation of Form I. For example, in one embodiment, the
appropriate solvent may be water or an alcohol/water mixture such
as 50% methanol/water mixture, even though the HCl salt form of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid is only sparingly soluble in water.
In one embodiment, the appropriate solvent is water.
[0222] The effective amount of time for formation of Form I from
the salt of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-
-3-methylpyridin-2-yl)benzoic acid can be any time between 2 to 24
hours or greater. It is recognized that the amount of time needed
is inversely proportional to the temperature. That is, the higher
the temperature the less time needed to affect dissociation of acid
to form Form I. When the solvent is water, stirring the dispersion
for approximately 24 hours at room temperature provides Form I in
an approximately 98% yield. If a solution of the salt of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid is desired for process purposes, an
elevated temperature may be used. After stirring the solution for
an effective amount of time at the elevated temperature,
recrystallization upon cooling provides substantially pure Form 1.
In one embodiment, substantially pure refers to greater than about
90% purity. In another embodiment, substantially pure refers to
greater than about 95% purity. In another embodiment, substantially
pure refers to greater than about 98% purity. In another
embodiment, substantially pure refers to greater than about 99%
purity. The temperature selected depends in part on the solvent
used and is well within the determination capabilities of one of
ordinary skill in the art. In one embodiment, the temperature is
between room temperature and about 80.degree. C. In another
embodiment, the temperature is between room temperature and about
40.degree. C. In another embodiment, the temperature is between
about 40.degree. C. and about 60.degree. C. In another embodiment,
the temperature is between about 60.degree. C. and about 80.degree.
C.
[0223] Compound 1 Form I may also be formed directly from
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)-t-butylbenzoate (cf. Scheme 3), which is a
precursor to the salt of Compound 1. Thus,
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)-t-butylbenzoate is allowed to undergo reaction
with an appropriate acid, such as, for example, formic acid under
appropriate reaction conditions to give Compound 1 Form I.
[0224] Compound 1 Form I may be further purified by
recrystallization from an organic solvent. Examples of organic
solvents include, but are not limited to, toluene, cumene, anisol,
1-butanol, isopropyl acetate, butyl acetate, isobutyl acetate,
methyl t-butyl ether, methyl isobutyl ketone and 1-propanol-water
mixtures. The temperature may be as described above. For example,
Form I is dissolved in 1-butanol at 75.degree. C. until it is
completely dissolved. Cooling down the solution to 10.degree. C. at
a rate of 0.2.degree. C./min yields crystals of Form I which may be
isolated by filtration.
[0225] In one embodiment, Compound 1 Form I is characterized by one
or more peaks at 15.2 to 15.6 degrees, 16.1 to 16.5 degrees, and
14.3 to 14.7 degrees in an X-ray powder diffraction obtained using
Cu K alpha radiation. In another embodiment, Compound 1 Form I is
characterized by one or more peaks at 15.4, 16.3, and 14.5 degrees.
In another embodiment, Compound 1 Form I is further characterized
by a peak at 14.6 to 15.0 degrees. In another embodiment, Compound
1 Form I is further characterized by a peak at 14.8 degrees. In
another embodiment, Compound 1 Form I is further characterized by a
peak at 17.6 to 18.0 degrees. In another embodiment, Compound 1
Form I is further characterized by a peak at 17.8 degrees. In
another embodiment, Compound 1 Form I is further characterized by a
peak at 16.4 to 16.8 degrees. In another embodiment, Compound 1
Form I is further characterized by a peak at 16.4 to 16.8 degrees.
In another embodiment, Compound 1 Form I is further characterized
by a peak at 16.6 degrees. In another embodiment, Compound 1 Form I
is further characterized by a peak at 7.6 to 8.0 degrees. In
another embodiment, Compound 1 Form I is further characterized by a
peak at 7.8 degrees. In another embodiment, Compound 1 Form I is
further characterized by a peak at 25.8 to 26.2 degrees. In another
embodiment, Compound 1 Form I is further characterized by a peak at
26.0 degrees. In another embodiment, Compound 1 Form I is further
characterized by a peak at 21.4 to 21.8 degrees. In another
embodiment, Compound 1 Form I is further characterized by a peak at
21.6 degrees. In another embodiment, Compound 1 Form I is further
characterized by a peak at 23.1 to 23.5 degrees. In another
embodiment, Compound 1 Form I is further characterized by a peak at
23.3 degrees. In some embodiments, Compound 1 Form I is
characterized by a diffraction pattern substantially similar to
that of FIG. 1. In some embodiments, Compound 1 Form I is
characterized by a diffraction pattern substantially similar to
that of FIG. 2.
[0226] In some embodiments, the particle size distribution of D90
is about 82 .mu.m or less for Compound 1 Form I. In some
embodiments, the particle size distribution of D50 is about 30
.mu.m or less for Compound 1 Form I.
Compound 1 Form II
[0227] Compound 1 Form II is prepared by slurrying Compound 1 Form
I in an appropriate solvent at a sufficient concentration for a
sufficient time. The slurry is then filtered centrifugally or under
vacuum and dried at ambient conditions for sufficient time to yield
Compound 1 Form II.
[0228] In some embodiments, about 20 to 40 mg of Compound 1 Form I
is slurried in about 400 to 600 .mu.L of an appropriate solvent. In
another embodiment, about 25 to 35 mg of Compound 1 Form I is
slurried in about 450 to 550 .mu.L of an appropriate solvent. In
another embodiment, about 30 mg of Compound 1 Form I is slurried in
about 500 .mu.L of an appropriate solvent.
[0229] In some embodiments, the time that Compound 1 Form I is
allowed to slurry with the solvent is from 1 hour to four days.
More particularly, the time that Compound 1 Form I is allowed to
slurry with the solvent is from 1 to 3 days. More particularly, the
time is 2 days.
[0230] In some embodiments, the appropriate solvent is selected
from an organic solvent of sufficient size to fit the voids in the
crystalline lattice of Compound 1 Form II. In other embodiments,
the solvate is of sufficient size to fit in voids measuring about
100 .ANG..sup.3.
[0231] In other embodiments, the solvent is selected from the group
consisting of methanol, ethanol, acetone, 2-propanol, acetonitrile,
tetrahydrofuran, methyl acetate, 2-butanone, ethyl formate, and
2-methyl tetrahydrofuran.
[0232] In other embodiments, a mixture of two or more of these
solvents may be used to obtain Compound 1 Form II. Alternatively,
Compound 1 Form II may be obtained from a mixture comprising one or
more of these solvents and water.
[0233] In some embodiments, the effective amount of time for drying
Compound 1 Form II is 1 to 24 hours. More particularly, the time is
6 to 18 hours. More particularly, the time is about 12 hours.
[0234] In another embodiment, Compound 1 Form II is prepared by
dispersing or dissolving a salt form of Compound 1, such as an HCl
salt of Compound 1 in an appropriate solvent for an effective
amount of time.
[0235] Compound 1 Form II as disclosed herein comprises a
crystalline lattice of Compound 1 in which voids in the crystalline
lattice are empty, or occupied, or partially occupied by one or
more molecules of a suitable solvent. Suitable solvents include,
but are not limited to, methanol, ethanol, acetone, 2-propanol,
acetonitrile, tetrahydrofuran, methyl acetate, 2-butanone, ethyl
formate, and 2-methyl tetrahydrofuran. Certain physical
characteristics of Compound 1 isostructural solvate forms, such as
X-ray powder diffraction, melting point and DSC, are not
substantially affected by the particular solvent molecule in
question.
[0236] In one embodiment, Compound 1 Form II is characterized by
one or more peaks at 21.50 to 21.90 degrees, 8.80 to 9.20 degrees,
and 10.80 to 11.20 degrees in an X-ray powder diffraction obtained
using Cu K alpha radiation. In another embodiment, Compound 1 Form
II is characterized by one or more peaks at 21.50 to 21.90 degrees,
8.80 to 9.20 degrees, 10.80 to 11.20 degrees, 18.00 to 18.40
degrees, and 22.90 to 23.30 degrees in an X-ray powder diffraction
obtained using Cu K alpha radiation. In another embodiment,
Compound 1 Form II is characterized by one or more peaks at 21.70,
8.98, and 11.04 degrees. In another embodiment, Compound 1 Form II
is characterized by one or more peaks at 21.70, 8.98, 11.04, 18.16,
and 23.06 degrees. In another embodiment, Compound 1 Form II is
characterized by a peak at 21.50 to 21.90 degrees. In another
embodiment, Compound 1 Form II is further characterized by a peak
at 21.70 degrees. In another embodiment, Compound 1 Form II is
further characterized by a peak at 8.80 to 9.20 degrees. In another
embodiment, Compound 1 Form II is further characterized by a peak
at 8.98 degrees. In another embodiment, Compound 1 Form II is
further characterized by a peak at 10.80 to 11.20 degrees. In
another embodiment, Compound 1 Form II is further characterized by
a peak at 11.04. In another embodiment, Compound 1 Form II is
further characterized by a peak at 18.00 to 18.40 degrees. In
another embodiment, Compound 1 Form II is further characterized by
a peak at 18.16 degrees. In another embodiment, Compound 1 Form II
is further characterized by a peak at 22.90 to 23.30 degrees. In
another embodiment, Compound 1 Form II is further characterized by
a peak at 23.06 degrees. In another embodiment, Compound 1 Form II
is further characterized by a peak at 20.40 to 20.80 degrees. In
another embodiment, Compound 1 Form II is further characterized by
a peak at 20.63 degrees. In another embodiment, Compound 1 Form II
is further characterized by a peak at 22.00 to 22.40 degrees. In
another embodiment, Compound 1 Form II is further characterized by
a peak at 22.22 degrees. In another embodiment, Compound 1 Form II
is further characterized by a peak at 18.40 to 18.80 degrees. In
another embodiment, Compound 1 Form II is further characterized by
a peak at 18.57 degrees. In another embodiment, Compound 1 Form II
is further characterized by a peak at 16.50 to 16.90 degrees. In
another embodiment, Compound 1 Form II is further characterized by
a peak at 16.66 degrees. In another embodiment, Compound 1 Form II
is further characterized by a peak at 19.70 to 20.10 degrees. In
another embodiment, Compound 1 Form II is further characterized by
a peak at 19.86 degrees.
[0237] In some embodiments, Compound 1 Form II is characterized by
a diffraction pattern substantially similar to that of FIG. 3. In
some embodiments, Compound 1 Form II is characterized by
diffraction patterns substantially similar to those provided in
FIG. 4.
[0238] In another embodiment, the solvate that forms Compound 1
Form II is selected from the group consisting of methanol, ethanol,
acetone, 2-propanol, acetonitrile, tetrahydrofuran, methyl acetate,
2-butanone, ethyl formate, and 2-methyl tetrahydrofuran.
Diffraction patterns are provided for the following Compound 1 Form
II: methanol (FIG. 5), ethanol (FIG. 6), acetone (FIG. 7),
2-propanol (FIG. 8), acetonitrile (FIG. 9), tetrahydrofuran (FIG.
10), methyl acetate (FIG. 11), 2-butanone (FIG. 12), ethyl formate
(FIG. 13), and 2-methyletrahydrofuran (FIG. 14).
[0239] In another embodiment, the invention provides Compound 1
Form II which exhibits two or more phase transitions as determined
by DSC or a similar analytic method known to the skilled artisan.
In some embodiments, the DSC of Compound 1 Form II is substantially
similar to the DSC trace depicted in FIG. 15. In another embodiment
of this aspect, the DSC gives two phase transitions. In another
embodiment, the DSC gives three phase transitions. In another
embodiment, one of the phase transitions occurs between 200 and
207.degree. C. In another embodiment, one of the phase transitions
occurs between 204 and 206.degree. C. In another embodiment, one of
the phase transitions occurs between 183 and 190.degree. C. In
another embodiment, one of the phase transitions occurs between 185
and 187.degree. C. In another embodiment, the melting point of
Compound 1, Solvate Form A is between 183.degree. C. to 190.degree.
C. In another embodiment, the melting point of Compound 1, Solvate
Form A is between 185.degree. C. to 187.degree. C.
[0240] In another embodiment, Compound 1 Form II comprises 1 to 10
weight percent (wt. %) solvate as determined by TGA. In some
embodiments, the TGA of Compound 1 Form II is substantially similar
to the TGA trace depicted in FIG. 16. In another embodiment,
Compound 1 Form II comprises 2 to 5 wt. % solvate as determined by
TGA or a similar analytic method known to the skilled artisan.
[0241] In another embodiment, the conformation of Compound 1 Form
II acetone solvate is substantially similar to that depicted in
FIG. 17, which is based on single X-ray analysis.
[0242] In another embodiment, Compound 1 Form II acetone solvate
has a P2.sub.1/n space group, and the following unit cell
dimensions:
a=16.5235(10).ANG..alpha.=90.degree.
b=12.7425(8).ANG..beta.=103.736(4).degree.
c=20.5512(13).ANG..gamma.=90.degree..
Compound 1 HCl Salt Form A
[0243] Compound 1 HCl Salt Form A can be prepared from the HCl salt
of Compound 1, by dissolving the HCl salt of Compound 1 in a
minimum of solvent and removing the solvent by slow evaporation. In
another embodiment, the solvent is an alcohol. In another
embodiment, the solvent is ethanol. Slow evaporation is generally
carried out by impeding the evaporation of the solvent. For
example, in one embodiment, slow evaporation involves dissolving
the HCl salt of Compound 1 in a vial and covering the vial with
parafilm that contains a hole poked in it.
[0244] In one embodiment, Compound 1 HCl Salt Form A is
characterized by one or more peaks at 8.80 to 9.20 degrees, 17.30
to 17.70 degrees, and 18.20 to 18.60 degrees in an X-ray powder
diffraction obtained using Cu K alpha radiation. In another
embodiment, Compound 1 HCl Salt Form A is characterized by one or
more peaks at 8.80 to 9.20 degrees, 17.30 to 17.70 degrees, 18.20
to 18.60 degrees, 10.10 to 10.50, and 15.80 to 16.20 degrees in an
X-ray powder diffraction obtained using Cu K alpha radiation. In
another embodiment, Compound 1 HCl Salt Form A is characterized by
one or more peaks at 8.96, 17.51, and 18.45 degrees. In another
embodiment, Compound 1 HCl Salt Form A is characterized by one or
more peaks at 8.96, 17.51, 18.45. 10.33, and 16.01 degrees. In
another embodiment, Compound 1 HCl Salt Form A is characterized by
a peak at 8.80 to 9.20 degrees. In another embodiment, Compound 1
HCl Salt Form A is characterized by a peak at 8.96 degrees. In
another embodiment, Compound 1 HCl Salt Form A is further
characterized by a peak at 17.30 to 17.70 degrees. In another
embodiment, Compound 1 HCl Salt Form A is characterized by a peak
at 17.51 degrees. In another embodiment, Compound 1 HCl Salt Form A
is further characterized by a peak at 18.20 to 18.60 degrees. In
another embodiment, Compound 1 HCl Salt Form A is further
characterized by a peak at 18.45 degrees. In another embodiment,
Compound 1 HCl Salt Form A is further characterized by a peak at
10.10 to 10.50 degrees. In another embodiment, Compound 1 HCl Salt
Form A is further characterized by a peak at 10.33 degrees. In
another embodiment, Compound 1 HCl Salt Form A is further
characterized by a peak at 15.80 to 16.20 degrees. In another
embodiment, Compound 1 HCl Salt Form A is further characterized by
a peak at 16.01 degrees. In another embodiment, Compound 1 HCl Salt
Form A is further characterized by a peak at 11.70 to 12.10
degrees. In another embodiment, Compound 1 HCl Salt Form A is
further characterized by a peak at 11.94 degrees. In another
embodiment, Compound 1 HCl Salt Form A is further characterized by
a peak at 7.90 to 8.30 degrees. In another embodiment, Compound 1
HCl Salt Form A is further characterized by a peak at 8.14 degrees.
In another embodiment, Compound 1 HCl Salt Form A is further
characterized by a peak at 9.90 to 10.30 degrees. In another
embodiment, Compound 1 HCl Salt Form A is further characterized by
a peak at 10.10 degrees. In another embodiment, Compound 1 HCl Salt
Form A is further characterized by a peak at 16.40 to 16.80
degrees. In another embodiment, Compound 1 HCl Salt Form A is
further characterized by a peak at 16.55 degrees. In another
embodiment, Compound 1 HCl Salt Form A is further characterized by
a peak at 9.30 to 9.70 degrees. In another embodiment, Compound 1
HCl Salt Form A is further characterized by a peak at 9.54 degrees.
In another embodiment, Compound 1 HCl Salt Form A is further
characterized by a peak at 16.40 to 16.80 degrees. In another
embodiment, Compound 1 HCl Salt Form A is further characterized by
a peak at 16.55 degrees. In some embodiments, Compound 1 HCl Salt
Form A is characterized as a dimer as depicted in FIG. 18.
[0245] In some embodiments, Compound 1 HCl Salt Form A is
characterized by a diffraction pattern substantially similar to
that of FIG. 19.
[0246] In another embodiment, the invention features crystalline
Compound 1 HCl Salt Form A having a P.sup.-1 space group, and the
following unit cell dimensions:
a=10.2702(2).ANG..alpha.=67.0270(10).degree.
b=10.8782(2).ANG..beta.=66.1810(10).degree.
c=12.4821(3).ANG..gamma.=72.4760(10).degree..
Methods for Making the Pharmaceutical Compositions
[0247] The dosage unit forms of the invention can be produced by
compacting or compressing an admixture or composition, for example,
a powder or granules, under pressure to form a stable
three-dimensional shape (e.g., a tablet). As used herein, "tablet"
includes compressed pharmaceutical dosage unit forms of all shapes
and sizes, whether coated or uncoated.
[0248] The expression "dosage unit form" as used herein refers to a
physically discrete unit of agent appropriate for the patient to be
treated. In general, a compacted mixture has a density greater than
that of the mixture prior to compaction. A dosage unit form of the
invention can have almost any shape including concave and/or convex
faces, rounded or angled corners, and a rounded to rectilinear
shape. In some embodiments, the compressed dosage forms of the
invention comprise a rounded tablet having flat faces. The solid
pharmaceutical dosage forms of the invention can be prepared by any
compaction and compression method known by persons of ordinary
skill in the art of forming compressed solid pharmaceutical dosage
forms. In particular embodiments, the formulations provided herein
may be prepared using conventional methods known to those skilled
in the field of pharmaceutical formulation, as described, e.g., in
pertinent textbooks. See, e.g., Remington: The Science and Practice
of Pharmacy, 21st Ed., Lippincott Williams & Wilkins,
Baltimore, Md. (2003); Ansel et al., Pharmaceutical Dosage Forms
And Drug Delivery Systems, 7th Edition, Lippincott Williams &
Wilkins, (1999); The Handbook of Pharmaceutical Excipients,
4.sup.th edition, Rowe et al., Eds., American Pharmaceuticals
Association (2003); Gibson, Pharmaceutical Preformulation And
Formulation, CRC Press (2001), these references hereby incorporated
herein by reference in their entirety.
Granulation and Compression
[0249] In some embodiments, solid forms, including powders
comprising the active agent Compound 1 and the included
pharmaceutically acceptable excipients (e.g. filler, diluent,
disintegrant, surfactant, glidant, binder, lubricant, or any
combination thereof) can be subjected to a dry granulation process.
The dry granulation process causes the powder to agglomerate into
larger particles having a size suitable for further processing. Dry
granulation can improve the flowability of a mixture in order to be
able to produce tablets that comply with the demand of mass
variation or content uniformity.
[0250] Formulations as described herein may be produced using one
or more mixing and dry granulations steps. The order and the number
of the mixing and granulation steps do not seem to be critical.
However, at least one of the excipients and Compound 1 can be been
subject to dry granulation or wet high shear granulation before
compression into tablets. Dry granulation of Compound 1 and the
excipients made together prior to tablet compression seem,
surprisingly, to be a simple, inexpensive and efficient way of
providing close physical contact between the ingredients of the
present compositions and formulations and thus results in a tablet
formulation with good stability properties. Dry granulation can be
carried out by a mechanical process, which transfers energy to the
mixture without any use of any liquid substances (neither in the
form of aqueous solutions, solutions based on organic solutes, or
mixtures thereof) in contrast to wet granulation processes, also
contemplated herein. Generally, the mechanical process requires
compaction such as the one provided by roller compaction. An
example of an alternative method for dry granulation is
slugging.
[0251] In some embodiments, roller compaction is a granulation
process comprising highly intensive mechanical compacting of one or
more substances. In some embodiments, a pharmaceutical composition
comprising an admixture of powders is pressed, that is roller
compacted, between 2 counter rotating rollers to make a solid sheet
which is subsequently crushed in a sieve to form a particulate
matter. In this particulate matter, a close mechanical contact
between the ingredients can be obtained. An example of roller
compaction equipment is Minipactor.RTM. a Gerteis 3W-Polygran from
Gerteis Maschinen+Processengineering AG.
[0252] In some embodiments, tablet compression according to the
invention can occur without any use of any liquid substances
(neither in the form of aqueous solutions, solutions based on
organic solutes, or mixtures thereof), i.e. a dry granulation
process. In a typical embodiment the resulting core or tablet has a
compressive strength in the range of 1 to 15 kP; such as 1.5 to
12.5 kP, preferably in the range of 2 to 10 kP.
Brief Manufacturing Procedure
[0253] In some embodiments, the ingredients are weighed according
to the formula set herein. Next, all of the intragranular
ingredients are sifted and mixed well. The ingredients can be
lubricated with a suitable lubricant, for example, magnesium
stearate. The next step can comprise compaction/slugging of the
powder admixture and sized ingredients. Next, the compacted or
slugged blends are milled into granules and sifted to obtain the
desired size. Next, the granules can be further lubricated with,
for example, magnesium stearate. Next the granular composition of
the invention can be compressed on suitable punches into various
pharmaceutical formulations in accordance with the invention.
Optionally the tablets can be coated with a film, colorant or other
coating.
[0254] Another aspect of the invention provides a method for
producing a pharmaceutical composition comprising providing an
admixture of a composition comprising Compound 1 and one or more
excipients selected from: a filler, a diluent, a binder, a glidant,
a surfactant, a lubricant, a disintegrant, and compressing the
composition into a tablet having a dissolution of at least about
50% in about 30 minutes.
[0255] In another embodiment, a wet granulation process is
performed to yield the pharmaceutical formulation of the invention
from an admixture of powdered and liquid ingredients. For example,
a pharmaceutical composition comprising an admixture of a
composition comprising Compound 1 and one or more excipients
selected from: a filler, a diluent, a binder, a glidant, a
surfactant, a lubricant, a disintegrant, are weighed as per the
formula set herein. Next, all of the intragranular ingredients are
sifted and mixed in a high shear or low shear granulator using
water or water with a surfactant or water with a binder or water
with a surfactant and a binder to granulate the powder blend. A
fluid other than water can also be used with or without surfactant
and/or binder to granulate the powder blend. Next, the wet granules
can optionally be milled using a suitable mill. Next, water may
optionally be removed from the admixture by drying the ingredients
in any suitable manner. Next, the dried granules can optionally be
milled to the required size. Next, extra granular excipients can be
added by blending (for example a filler, a diluent, and a
disintegrant). Next, the sized granules can be further lubricated
with magnesium stearate and a disintegrant, for example,
croscarmellose sodium. Next the granular composition of the
invention can be sifted for sufficient time to obtain the correct
size and then compressed on suitable punches into various
pharmaceutical formulations in accordance with the invention.
Optionally, the tablets can be coated with a film, colorant or
other coating.
[0256] Each of the ingredients of this exemplary admixture is
described above and in the Examples below. Furthermore, the
admixture can comprise optional additives, such as, one or more
colorants, one or more flavors, and/or one or more fragrances as
described above and in the Examples below. In some embodiments, the
relative concentrations (e.g., wt %) of each of these ingredients
(and any optional additives) in the admixture are also presented
above and in the Examples below. The ingredients constituting the
admixture can be provided sequentially or in any combination of
additions; and, the ingredients or combination of ingredients can
be provided in any order. In one embodiment, the lubricant is the
last component added to the admixture.
[0257] In another embodiment, the admixture comprises a composition
of Compound 1, and any one or more of the excipients; a binder, a
glidant, a surfactant, a diluent, a lubricant, a disintegrant, and
a filler, wherein each of these ingredients is provided in a powder
form (e.g., provided as particles having a mean or average
diameter, measured by light scattering, of 250 .mu.m or less (e.g.,
150 .mu.m or less, 100 .mu.m or less, 50 .mu.m or less, 45 .mu.m or
less, 40 .mu.m or less, or 35 .mu.m or less)). For instance, the
admixture comprises a composition of Compound 1, a diluent, a
glidant, a surfactant, a lubricant, a disintegrant, and a filler,
wherein each of these ingredients is provided in a powder form
(e.g., provided as particles having a mean diameter, measured by
light scattering, of 250 .mu.m or less (e.g., 150 .mu.m or less,
100 .mu.m or less, 50 .mu.m or less, 45 .mu.m or less, 40 .mu.m or
less, or 35 .mu.m or less)). In another example, the admixture
comprises a composition of Compound 1, a diluent, a binder, a
surfactant, a lubricant, a disintegrant, and a filler, wherein each
of these ingredients is provided in a powder form (e.g., provided
as particles having a mean diameter, measured by light scattering,
of 250 .mu.m or less (e.g., 150 .mu.m or less, 100 .mu.m or less,
50 .mu.m or less, 45 .mu.m or less, 40 .mu.m or less, or 35 .mu.m
or less))
[0258] In another embodiment, the admixture comprises a composition
of Compound 1, and any combination of: a binder, a glidant, a
diluent, a surfactant, a lubricant, a disintegrant, and a filler,
wherein each of these ingredients is substantially free of water.
Each of the ingredients comprises less than 5 wt % (e.g., less than
2 wt %, less than 1 wt %, less than 0.75 wt %, less than 0.5 wt %,
or less than 0.25 wt %) of water by weight of the ingredient. For
instance, the admixture comprises a composition of Compound 1, a
diluent, a glidant, a surfactant, a lubricant, a disintegrant, and
a filler, wherein each of these ingredients is substantially free
of water. In some embodiments, each of the ingredients comprises
less than 5 wt % (e.g., less than 2 wt %, less than 1 wt %, less
than 0.75 wt %, less than 0.5 wt %, or less than 0.25 wt %) of
water by weight of the ingredient.
[0259] In another embodiment, compressing the admixture into a
tablet is accomplished by filling a form (e.g., a mold) with the
admixture and applying pressure to admixture. This can be
accomplished using a die press or other similar apparatus. In some
embodiments, the admixture of Compound 1 and excipients can be
first processed into granular form. The granules can then be sized
and compressed into tablets or formulated for encapsulation
according to known methods in the pharmaceutical art. It is also
noted that the application of pressure to the admixture in the form
can be repeated using the same pressure during each compression or
using different pressures during the compressions. In another
example, the admixture of powdered ingredients or granules can be
compressed using a die press that applies sufficient pressure to
form a tablet having a dissolution of about 50% or more at about 30
minutes (e.g., about 55% or more at about 30 minutes or about 60%
or more at about 30 minutes). For instance, the admixture is
compressed using a die press to produce a tablet hardness of at
least about 5 kP (at least about 5.5 kP, at least about 6 kP, at
least about 7 kP, at least about 10 kP, or at least 15 kP). In some
instances, the admixture is compressed to produce a tablet hardness
of between about 5 and 200.
[0260] In some embodiments, tablets comprising a pharmaceutical
composition as described herein can be coated with about 3.0 wt %
of a film coating comprising a colorant by weight of the tablet. In
certain instances, the colorant suspension or solution used to coat
the tablets comprises about 20% w/w of solids by weight of the
colorant suspension or solution. In still further instances, the
coated tablets can be labeled with a logo, other image or text.
[0261] In another embodiment, the method for producing a
pharmaceutical composition comprises providing an admixture of a
solid forms, e.g. an admixture of powdered and/or liquid
ingredients, the admixture comprising Compound 1 and one or more
excipients selected from: a binder, a glidant, a diluent, a
surfactant, a lubricant, a disintegrant, and a filler; mixing the
admixture until the admixture is substantially homogenous, and
compressing or compacting the admixture into a granular form. Then
the granular composition comprising Compound 1 can be compressed
into tablets or formulated into capsules as described above or in
the Examples below. Alternatively, methods for producing a
pharmaceutical composition comprises providing an admixture of
Compound 1, and one or more excipients, e.g. a binder, a glidant, a
diluent, a surfactant, a lubricant, a disintegrant, and a filler;
mixing the admixture until the admixture is substantially
homogenous, and compressing/compacting the admixture into a
granular form using a roller compactor using a dry granulation
composition as set forth in the Examples below or alternatively,
compressed/compacted into granules using a high shear wet granule
compaction process as set forth in the Examples below.
Pharmaceutical formulations, for example a tablet as described
herein, can be made using the granules prepared incorporating
Compound 1 in addition to the selected excipients described
herein.
[0262] In some embodiments, the admixture is mixed by stirring,
blending, shaking, or the like using hand mixing, a mixer, a
blender, any combination thereof, or the like. When ingredients or
combinations of ingredients are added sequentially, mixing can
occur between successive additions, continuously throughout the
ingredient addition, after the addition of all of the ingredients
or combinations of ingredients, or any combination thereof. The
admixture is mixed until it has a substantially homogenous
composition.
[0263] In another embodiment, the present invention comprises jet
milling Compound 1, Compound 1 Form I, Compound 1 Form II, Compound
1 HCl Salt Form A in a suitable, conventional milling apparatus
using air pressure suitable to produce particles having a
significant particle size fraction between 0.1 microns and 50
microns. In another embodiment, the particle size is between 0.1
microns and 20 microns. In another embodiment, the particles size
is between 0.1 microns and 10 microns. In another embodiment, the
particle size is between 1.0 microns and 5 microns. In still
another embodiment, Compound 1, Compound 1 Form I, Compound 1 Form
II, Compound 1 HCl Salt Form A has a particle size D50 of 2.0
microns.
[0264] In various embodiments, a second therapeutic agent can be
formulated together with Compound 1 to form a unitary or single
dose form, for example, a tablet or capsule.
[0265] Dosage forms prepared as above can be subjected to in vitro
dissolution evaluations according to Test 711 "Dissolution" in
United States Pharmacopoeia 29, United States Pharmacopeial
Convention, Inc., Rockville, Md., 2005 ("USP"), to determine the
rate at which the active substance is released from the dosage
forms. The content of active substance and the impurity levels are
conveniently measured by techniques such as high performance liquid
chromatography (HPLC).
[0266] In some embodiments, the invention includes use of packaging
materials such as containers and closures of high-density
polyethylene (HDPE), low-density polyethylene (LDPE) and or
polypropylene and/or glass, glassine foil, aluminum pouches, and
blisters or strips composed of aluminum or high-density polyvinyl
chloride (PVC), optionally including a desiccant, polyethylene
(PE), polyvinylidene dichloride (PVDC), PVC/PE/PVDC, and the like.
These package materials can be used to store the various
pharmaceutical compositions and formulations in a sterile fashion
after appropriate sterilization of the package and its contents
using chemical or physical sterilization techniques commonly
employed in the pharmaceutical arts.
Methods for Administering the Pharmaceutical Compositions
[0267] In one aspect, the pharmaceutical compositions of the
invention can be administered to a patient once daily or about
every twenty four hours. Alternatively, the pharmaceutical
compositions of the invention can be administered to a patient
twice daily or about every twelve hours. These pharmaceutical
compositions are administered as oral formulations containing about
25 mg, 50 mg, 100 mg, 125 mg, 150 mg, 200 mg, 250 mg, or 400 mg of
Compound 1. In this aspect, in addition to Compound 1, the
pharmaceutical compositions comprise a filler; a diluent; a
disintegrant; a surfactant; at least one of a binder and a glidant;
and a lubricant. For instance, a dose of 400 mg of Compound 1, may
comprise two tablets of the invention each containing 200 mg of
Compound 1, or four tablets of the invention each containing 100 mg
of Compound 1.
[0268] It will also be appreciated that the compound and
pharmaceutically acceptable compositions and formulations of the
invention can be employed in combination therapies; that is,
Compound 1 and pharmaceutically acceptable compositions thereof can
be administered concurrently with, prior to, or subsequent to, one
or more other desired therapeutics or medical procedures. The
particular combination of therapies (therapeutics or procedures) to
employ in a combination regimen will take into account
compatibility of the desired therapeutics and/or procedures and the
desired therapeutic effect to be achieved. It will also be
appreciated that the therapies employed may achieve a desired
effect for the same disorder (for example, an inventive compound
may be administered concurrently with another agent used to treat
the same disorder), or they may achieve different effects (e.g.,
control of any adverse effects). As used herein, additional
therapeutic agents that are normally administered to treat or
prevent a particular disease, for example, a CFTR mediated disease,
or condition, are known as "appropriate for the disease or
condition being treated."
[0269] In one embodiment, the additional therapeutic agent is
selected from a mucolytic agent, bronchodialator, an antibiotic, an
anti-infective agent, an anti-inflammatory agent, a CFTR modulator
other than Compound 1 of the invention, or a nutritional agent.
[0270] In one embodiment, the additional agent is
(R)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-N-(1-(2,3-dihydroxypropyl)-6-
-fluoro-2-(1-hydroxy-2-methylpropan-2-yl)-1H-indol-5-yl)cyclopropanecarbox-
amide. In another embodiment, the additional agent is
N-(5-hydroxy-2,4-ditert-butyl-phenyl)-4-oxo-1H-quinoline-3-carboxamide.
In another embodiment, the additional agent is selected from Table
1:
TABLE-US-00012 TABLE 1 ##STR00006## 1 ##STR00007## 2 ##STR00008## 3
##STR00009## 4 ##STR00010## 5 ##STR00011## 6 ##STR00012## 7
##STR00013## 8 ##STR00014## 9 ##STR00015## 10 ##STR00016## 11
##STR00017## 12 ##STR00018## 13 ##STR00019## 14
[0271] In another embodiment, the additional agent is any
combination of the above agents. For example, the composition may
comprise Compound 1,
(R)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-N-(1-(2,3-dihydroxypropyl)-6-
-fluoro-2-(1-hydroxy-2-methylpropan-2-yl)-1H-indol-5-yl)cyclopropanecarbox-
amide, and
N-(5-hydroxy-2,4-ditert-butyl-phenyl)-4-oxo-1H-quinoline-3-carb-
oxamide. In another example, the composition may comprise Compound
1,
N-(5-hydroxy-2,4-ditert-butyl-phenyl)-4-oxo-1H-quinoline-3-carboxamide,
and any one of the compounds from Table 1, i.e. compounds 1 through
14 of Table 1, or any combination thereof.
[0272] In one embodiment, the additional therapeutic agent is an
antibiotic. Exemplary antibiotics useful herein include tobramycin,
including tobramycin inhaled powder (TIP), azithromycin, aztreonam,
including the aerosolized form of aztreonam, amikacin, including
liposomal formulations thereof, ciprofloxacin, including
formulations thereof suitable for administration by inhalation,
levoflaxacin, including aerosolized formulations thereof, and
combinations of two antibiotics, e.g., fosfomycin and
tobramycin.
[0273] In another embodiment, the additional agent is a mucolyte.
Exemplary mucolytes useful herein includes Pulmozyme.RTM..
[0274] In another embodiment, the additional agent is a
bronchodialator. Exemplary bronchodialtors include albuterol,
metaprotenerol sulfate, pirbuterol acetate, salmeterol, or
tetrabuline sulfate.
[0275] In another embodiment, the additional agent is effective in
restoring lung airway surface liquid. Such agents improve the
movement of salt in and out of cells, allowing mucus in the lung
airway to be more hydrated and, therefore, cleared more easily.
Exemplary such agents include hypertonic saline, denufosol
tetrasodium
([[(3S,5R)-5-(4-amino-2-oxopyrimidin-1-yl)-3-hydroxyoxolan-2-yl]methoxy-h-
ydroxyphosphoryl][[[(2R,3S,4R,5R)-5-(2,4-dioxopyrimidin-1-yl)-3,4-dihydrox-
yoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-hydroxyphosphoryl]hydrogen
phosphate), or bronchitol (inhaled formulation of mannitol).
[0276] In another embodiment, the additional agent is an
anti-inflammatory agent, i.e., an agent that can reduce the
inflammation in the lungs. Exemplary such agents useful herein
include ibuprofen, docosahexanoic acid (DHA), sildenafil, inhaled
glutathione, pioglitazone, hydroxychloroquine, or simavastatin.
[0277] In another embodiment, the additional agent is a CFTR
modulator other than Compound 1, i.e., an agent that has the effect
of modulating CFTR activity. Exemplary such agents include ataluren
("PTC124.RTM."; 3-[5-(2-fluorophenyl)-1,2,4-oxadiazol-3-yl]benzoic
acid), sinapultide, lancovutide, depelestat (a human recombinant
neutrophil elastase inhibitor), and cobiprostone
(7-{(2R,4aR,5R,7aR)-2-[(3S)-1,1-difluoro-3-methylpentyl]-2-hydroxy-6-oxoo-
ctahydrocyclopenta[b]pyran-5-yl}heptanoic acid).
[0278] In another embodiment, the additional agent is a nutritional
agent. Exemplary nutritional agents include pancrelipase
(pancreating enzyme replacement), including Pancrease.RTM.,
Pancreacarb.RTM., Ultrase.RTM., or Creon.RTM., Liprotomase.RTM.
(formerly Trizytek.RTM.), Aquadeks.RTM., or glutathione inhalation.
In one embodiment, the additional nutritional agent is
pancrelipase.
[0279] In another embodiment, the additional agent is a compound
selected from gentamicin, curcumin, cyclophosphamide,
4-phenylbutyrate, miglustat, felodipine, nimodipine, Philoxin B,
geniestein, Apigenin, cAMP/cGMP modulators such as rolipram,
sildenafil, milrinone, tadalafil, aminone, isoproterenol,
albuterol, and almeterol, deoxyspergualin, HSP 90 inhibitors, HSP
70 inhibitors, proteosome inhibitors such as epoxomicin,
lactacystin, etc.
[0280] In other embodiments, the additional agent is a compound
disclosed in WO 2004028480, WO 2004110352, WO 2005094374, WO
2005120497, or WO 2006101740. In another embodiment, the additional
agent is a benzo[c]quinolizinium derivative that exhibits CFTR
modulation activity or a benzopyran derivative that exhibits CFTR
modulation activity. In another embodiment, the additional agent is
a compound disclosed in U.S. Pat. No. 7,202,262, U.S. Pat. No.
6,992,096, US20060148864, US20060148863, US20060035943,
US20050164973, WO2006110483, WO2006044456, WO2006044682,
WO2006044505, WO2006044503, WO2006044502, or WO2004091502. In
another embodiment, the additional agent is a compound disclosed in
WO2004080972, WO2004111014, WO2005035514, WO2005049018,
WO2006099256, WO2006127588, or WO2007044560. In another embodiment,
the additional agent is
N-(5-hydroxy-2,4-ditert-butyl-phenyl)-4-oxo-1H-quinoline-3-carbo-
xamide.
[0281] In one embodiment, 400 mg of Compound 1 may be administered
to a subject in need thereof followed by co-administration of 150
mg of
N-(5-hydroxy-2,4-ditert-butyl-phenyl)-4-oxo-1H-quinoline-3-carboxamide
(Compound 2). In another embodiment, 400 mg of Compound 1 may be
administered to a subject in need thereof followed by
co-administration of 250 mg of Compound 2. In these embodiments,
the dosage amounts may be achieved by administration of one or more
tablets of the invention. For example, administration of 400 mg of
Compound 1 may be achieved by administering two tablets each
containing 200 mg of Compound 1, or four tablets each containing
100 mg of Compound 1. Compound 2 may be administered as a
pharmaceutical composition comprising Compound 2 and a
pharmaceutically acceptable carrier. The duration of administration
may continue until amelioration of the disease is achieved or until
a subject's physician advises, e.g. duration of administration may
be less than a week, 1 week, 2 weeks, 3 weeks, or a month or
longer. The co-administration period may be preceded by an
administration period of just Compound 1 alone. For example, there
could be administration of 400 mg of Compound 1 for 2 weeks
followed by co-administration of 150 mg or 250 mg of Compound 2 for
1 additional week.
[0282] In one embodiment, 400 mg of Compound 1 may be administered
once a day to a subject in need thereof followed by
co-administration of 150 mg of Compound 2 once a day. In another
embodiment, 400 mg of Compound 1 may be administered once a day to
a subject in need thereof followed by co-administration of 250 mg
of Compound 2 once a day. In these embodiments, the dosage amounts
may be achieved by administration of one or more tablets of the
invention. For example, administration of 400 mg of Compound 1 may
be achieved by administering two tablets each containing 200 mg of
Compound 1, or four tablets each containing 100 mg of Compound 1.
Compound 2 may be administered as a pharmaceutical composition
comprising Compound 2 and a pharmaceutically acceptable carrier.
The duration of administration may continue until amelioration of
the disease is achieved or until a subject's physician advises,
e.g. duration of administration may be less than a week, 1 week, 2
weeks, 3 weeks, or a month or longer. The co-administration period
may be preceded by an administration period of just Compound 1
alone. For example, there could be administration of 400 mg of
Compound 1 for 2 weeks followed by co-administration of 150 mg or
250 mg of Compound 2 for 1 additional week.
[0283] In one embodiment, 400 mg of Compound 1 may be administered
once a day to a subject in need thereof followed by
co-administration of 150 mg of Compound 2 every 12 hours. In
another embodiment, 400 mg of Compound 1 may be administered once a
day to a subject in need thereof followed by co-administration of
250 mg of Compound 2 every 12 hours. In these embodiments, the
dosage amounts may be achieved by administration of one or more
tablets of the invention. For example, administration of 400 mg of
Compound 1 may be achieved by administering two tablets each
containing 200 mg of Compound 1, or four tablets each containing
100 mg of Compound 1. Compound 2 may be administered as a
pharmaceutical composition comprising Compound 2 and a
pharmaceutically acceptable carrier. The duration of administration
may continue until amelioration of the disease is achieved or until
a subject's physician advises, e.g. duration of administration may
be less than a week, 1 week, 2 weeks, 3 weeks, or a month or
longer. The co-administration period may be preceded by an
administration period of just Compound 1 alone. For example, there
could be administration of 400 mg of Compound 1 for 2 weeks
followed by co-administration of 150 mg or 250 mg of Compound 2 for
1 additional week.
[0284] These combinations are useful for treating the diseases
described herein including cystic fibrosis. These combinations are
also useful in the kits described herein.
[0285] The amount of additional therapeutic agent present in the
compositions of this invention will be no more than the amount that
would normally be administered in a composition comprising that
therapeutic agent as the only active agent. Preferably the amount
of additional therapeutic agent in the presently disclosed
compositions will range from about 50% to 100% of the amount
normally present in a composition comprising that agent as the only
therapeutically active agent.
Therapeutic Uses of the Composition
[0286] In certain embodiments, the pharmaceutically acceptable
compositions comprising Compound 1 and optionally an additional
agent are useful for treating or lessening the severity of cystic
fibrosis in patients who exhibit residual CFTR activity in the
apical membrane of respiratory and non-respiratory epithelia. The
presence of residual CFTR activity at the epithelial surface can be
readily detected using methods known in the art, e.g., standard
electrophysiological, biochemical, or histochemical techniques.
Such methods identify CFTR activity using in vivo or ex vivo
electrophysiological techniques, measurement of sweat or salivary
Cl.sup.- concentrations, or ex vivo biochemical or histochemical
techniques to monitor cell surface density. Using such methods,
residual CFTR activity can be readily detected in patients
heterozygous or homozygous for a variety of different mutations,
including patients homozygous or heterozygous for the most common
mutation, .DELTA.F508, as well as other mutations such as the G551D
mutation, or the R117H mutation.
[0287] In one embodiment, Compound 1, as described herein, or
pharmaceutically acceptable compositions thereof, are useful for
treating or lessening the severity of cystic fibrosis in patients
within certain genotypes exhibiting residual CFTR activity, e.g.,
class III mutations (impaired regulation or gating), class IV
mutations (altered conductance), or class V mutations (reduced
synthesis) (Lee R. Choo-Kang, Pamela L., Zeitlin, Type I, II, III,
IV, and V cystic fibrosis Tansmembrane Conductance Regulator
Defects and Opportunities of Therapy; Current Opinion in Pulmonary
Medicine 6:521-529, 2000). Other patient genotypes that exhibit
residual CFTR activity include patients homozygous for one of these
classes or heterozygous with any other class of mutations,
including class I mutations, class II mutations, or a mutation that
lacks classification.
[0288] In one embodiment, Compound 1, as described herein, or
pharmaceutically acceptable compositions thereof, are useful for
treating or lessening the severity of cystic fibrosis in patients
within certain clinical phenotypes, e.g., a moderate to mild
clinical phenotype that typically correlates with the amount of
residual CFTR activity in the apical membrane of epithelia. Such
phenotypes include patients exhibiting pancreatic insufficiency or
patients diagnosed with idiopathic pancreatitis and congenital
bilateral absence of the vas deferens, or mild lung disease.
[0289] The exact amount required will vary from subject to subject,
depending on the species, age, and general condition of the
subject, the severity of the infection, the particular agent, its
mode of administration, and the like. The compounds of the
invention are preferably formulated in dosage unit form for ease of
administration and uniformity of dosage. The expression "dosage
unit form" as used herein refers to a physically discrete unit of
agent appropriate for the patient to be treated. It will be
understood, however, that the total daily usage of the compounds
and compositions of the invention will be decided by the attending
physician within the scope of sound medical judgment. The specific
effective dose level for any particular patient or organism will
depend upon a variety of factors including the disorder being
treated and the severity of the disorder; the activity of the
specific compound employed; the specific composition employed; the
age, body weight, general health, sex and diet of the patient; the
time of administration, route of administration, and rate of
excretion of the specific compound employed; the duration of the
treatment; drugs used in combination or coincidental with the
specific compound employed, and like factors well known in the
medical arts. The term "patient", as used herein, means an animal,
preferably a mammal, and most preferably a human.
[0290] Anywhere in the present application where a name of a
compound may not correctly describe the structure of the compound,
the structure supersedes the name and governs.
EXAMPLES
[0291] XRPD (X-Ray Powder Diffraction)
[0292] The X-Ray diffraction (XRD) data of Compound 1, Compound 1
Form I, Compound 1 Form II, or Compound 1 HCl Salt Form A were
collected on a Bruker D8 DISCOVER powder diffractometer with
HI-STAR 2-dimensional detector and a flat graphite monochromator.
Cu sealed tube with K.alpha. radiation was used at 40 kV, 35 mA.
The samples were placed on zero-background silicon wafers at
25.degree. C. For each sample, two data frames were collected at
120 seconds each at 2 different .theta..sub.2 angles: 8.degree. and
26.degree.. The data were integrated with GADDS software and merged
with DIFFRACTP.sup.plusEVA software. Uncertainties for the reported
peak positions are .+-.0.2 degrees.
[0293] Jet Milling Description
[0294] Unmicronized Compound 1, Compound 1 Form I, Compound 1 Form
II, or Compound 1 HCl Salt Form A is sieved to de-lump it prior to
placing it into the jet mill hopper. All sieves are disposable and
received a wipe prior to use. Unmicronized Compound 1, Compound 1
Form I, Compound 1 Form II, or Compound 1 HCl Salt Form A is added
to the jet mill hopper at a controlled feeding rate using
compressed nitrogen gas. The gas pressure range is 40-45/45-70
(Venturi/Mill) PSI and the feeding rate range is 0.5-1.6 Kg/Hour.
The Compound 1, Compound 1 Form I, Compound 1 Form II, or Compound
1 HCl Salt Form A is micronized in the mill through
particle-particle and particle-wall collisions and the processed
Compound 1, Compound 1 Form I, Compound 1 Form II, or Compound 1
HCl Salt Form A is emptied into the micronized product containers.
It is believed that one of ordinary skill in the art may also
achieve Compound 1, Compound 1 Form I, Compound 1 Form II, or
Compound 1 HCl Salt Form A with a favorable particle size through
pin milling based in part on the conditions described above.
[0295] Differential Scanning Calorimetry (DSC)
[0296] The Differential scanning calorimetry (DSC) data of Compound
1, Compound 1 Form I, Compound 1 Form II, or Compound 1 HCl Salt
Form A were collected using a DSC Q100 V9.6 Build 290 (TA
Instruments, New Castle, Del.). Temperature was calibrated with
indium and heat capacity was calibrated with sapphire. Samples of
3-6 mg were weighed into aluminum pans that were crimped using lids
with 1 pin hole. The samples were scanned from 25.degree. C. to
350.degree. C. at a heating rate of 1.0.degree. C./min and with a
nitrogen gas purge of 50 ml/min. Data were collected by Thermal
Advantage Q Series.TM. version 2.2.0.248 software and analyzed by
Universal Analysis software version 4.1D (TA Instruments, New
Castle, Del.). The reported numbers represent single analyses.
[0297] Compound 1 Form I, Compound 1 Form II, and Compound 1 HCl
Salt Form A Single Crystal Structure Determination
[0298] Diffraction data were acquired on Bruker Apex II
diffractometer equipped with sealed tube Cu K-alpha source and an
Apex II CCD detector. The structure was solved and refined using
SHELX program (Sheldrick, G. M., Acta Cryst., (2008) A64, 112-122).
Based on systematic absences and intensities statistics the
structure was solved and refined in P2.sub.1/n space group.
[0299] Vitride.RTM. (sodium bis(2-methoxyethoxy)aluminum hydride
[or NaAlH.sub.2(OCH.sub.2CH.sub.2OCH.sub.3).sub.2], 65 wgt %
solution in toluene) was purchased from Aldrich Chemicals.
[0300] 2,2-Difluoro-1,3-benzodioxole-5-carboxylic acid was
purchased from Saltigo (an affiliate of the Lanxess
Corporation).
Preparation of (2,2-difluoro-1,3-benzodioxol-5-yl)-methanol
##STR00020##
[0302] Commercially available
2,2-difluoro-1,3-benzodioxole-5-carboxylic acid (1.0 eq) was
slurried in toluene (10 vol). Vitride.RTM. (2 eq) was added via
addition funnel at a rate to maintain the temperature at
15-25.degree. C. At the end of the addition, the temperature was
increased to 40.degree. C. for 2 hours (h), then 10% (w/w) aqueous
(aq) NaOH (4.0 eq) was carefully added via addition funnel,
maintaining the temperature at 40-50.degree. C. After stirring for
an additional 30 minutes (min), the layers were allowed to separate
at 40.degree. C. The organic phase was cooled to 20.degree. C.,
then washed with water (2.times.1.5 vol), dried (Na.sub.2SO.sub.4),
filtered, and concentrated to afford crude
(2,2-difluoro-1,3-benzodioxol-5-yl)-methanol that was used directly
in the next step.
Preparation of 5-chloromethyl-2,2-difluoro-1,3-benzodioxole
##STR00021##
[0304] (2,2-difluoro-1,3-benzodioxol-5-yl)-methanol (1.0 eq) was
dissolved in MTBE (5 vol). A catalytic amount of
4-(N,N-dimethyl)aminopyridine (DMAP) (1 mol %) was added and
SOCl.sub.2 (1.2 eq) was added via addition funnel. The SOCl.sub.2
was added at a rate to maintain the temperature in the reactor at
15-25.degree. C. The temperature was increased to 30.degree. C. for
1 h, and then was cooled to 20.degree. C. Water (4 vol) was added
via addition funnel while maintaining the temperature at less than
30.degree. C. After stirring for an additional 30 min, the layers
were allowed to separate. The organic layer was stirred and 10%
(w/v) aq NaOH (4.4 vol) was added. After stirring for 15 to 20 min,
the layers were allowed to separate. The organic phase was then
dried (Na.sub.2SO.sub.4), filtered, and concentrated to afford
crude 5-chloromethyl-2,2-difluoro-1,3-benzodioxole that was used
directly in the next step.
Preparation of (2,2-difluoro-1,3-benzodioxol-5-yl)-acetonitrile
##STR00022##
[0306] A solution of 5-chloromethyl-2,2-difluoro-1,3-benzodioxole
(1 eq) in DMSO (1.25 vol) was added to a slurry of NaCN (1.4 eq) in
DMSO (3 vol), while maintaining the temperature between
30-40.degree. C. The mixture was stirred for 1 h, and then water (6
vol) was added, followed by methyl tert-butyl ether (MTBE) (4 vol).
After stirring for 30 min, the layers were separated. The aqueous
layer was extracted with MTBE (1.8 vol). The combined organic
layers were washed with water (1.8 vol), dried (Na.sub.2SO.sub.4),
filtered, and concentrated to afford crude
(2,2-difluoro-1,3-benzodioxol-5-yl)-acetonitrile (95%) that was
used directly in the next step.
Synthesis of
(2,2-difluoro-1,3-benzodioxol-5-yl)-1-ethylacetate-acetonitrile
##STR00023##
[0308] A reactor was purged with nitrogen and charged with 900 mL
of toluene. The solvent was degassed via nitrogen sparge for no
less than 16 h. To the reactor was then charged Na.sub.3PO.sub.4
(155.7 g, 949.5 mmol), followed by bis(dibenzylideneacetone)
palladium (0) (7.28 g, 12.66 mmol). A 10% w/w solution of
tert-butylphosphine in hexanes (51.23 g, 25.32 mmol) was charged
over 10 min at 23.degree. C. from a nitrogen purged addition
funnel. The mixture was allowed to stir for 50 min, at which time
5-bromo-2,2-difluoro-1,3-benzodioxole (75 g, 316.5 mmol) was added
over 1 min. After stirring for an additional 50 min, the mixture
was charged with ethyl cyanoacetate (71.6 g, 633.0 mmol) over 5 min
followed by water (4.5 mL) in one portion. The mixture was heated
to 70.degree. C. over 40 min and analyzed by HPLC every 1-2 h for
the percent conversion of the reactant to the product. After
complete conversion was observed (typically 100% conversion after
5-8 h), the mixture was cooled to 20-25.degree. C. and filtered
through a celite pad. The celite pad was rinsed with toluene
(2.times.450 mL) and the combined organics were concentrated to 300
mL under vacuum at 60-65.degree. C. The concentrate was charged
with 225 mL DMSO and concentrated under vacuum at 70-80.degree. C.
until active distillation of the solvent ceased. The solution was
cooled to 20-25.degree. C. and diluted to 900 mL with DMSO in
preparation for Step 2. .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
7.16-7.10 (m, 2H), 7.03 (d, J=8.2 Hz, 1H), 4.63 (s, 1H), 4.19 (m,
2H), 1.23 (t, J=7.1 Hz, 3H).
Synthesis of (2,2-difluoro-1,3-benzodioxol-5-yl)-acetonitrile
##STR00024##
[0310] The DMSO solution of
(2,2-difluoro-1,3-benzodioxol-5-yl)-1-ethylacetate-acetonitrile
from above was charged with 3 N HCl (617.3 mL, 1.85 mol) over 20
min while maintaining an internal temperature<40.degree. C. The
mixture was then heated to 75.degree. C. over 1 h and analyzed by
HPLC every 1-2 h for % conversion. When a conversion of >99% was
observed (typically after 5-6 h), the reaction was cooled to
20-25.degree. C. and extracted with MTBE (2.times.525 mL), with
sufficient time to allow for complete phase separation during the
extractions. The combined organic extracts were washed with 5% NaCl
(2.times.375 mL). The solution was then transferred to equipment
appropriate for a 1.5-2.5 Torr vacuum distillation that was
equipped with a cooled receiver flask. The solution was
concentrated under vacuum at <60.degree. C. to remove the
solvents. (2,2-Difluoro-1,3-benzodioxol-5-yl)-acetonitrile was then
distilled from the resulting oil at 125-130.degree. C. (oven
temperature) and 1.5-2.0 Torr.
(2,2-Difluoro-1,3-benzodioxol-5-yl)-acetonitrile was isolated as a
clear oil in 66% yield from 5-bromo-2,2-difluoro-1,3-benzodioxole
(2 steps) and with an HPLC purity of 91.5% AUC (corresponds to a
w/w assay of 95%). .sup.1H NMR (500 MHz, DMSO) .delta. 7.44 (br s,
1H), 7.43 (d, J=8.4 Hz, 1H), 7.22 (dd, J=8.2, 1.8 Hz, 1H), 4.07 (s,
2H).
Preparation of
(2,2-difluoro-1,3-benzodioxol-5-yl)-cyclopropanecarbonitrile
##STR00025##
[0312] A mixture of
(2,2-difluoro-1,3-benzodioxol-5-yl)-acetonitrile (1.0 eq), 50 wt %
aqueous KOH (5.0 eq) 1-bromo-2-chloroethane (1.5 eq), and
Oct.sub.4NBr (0.02 eq) was heated at 70.degree. C. for 1 h. The
reaction mixture was cooled, then worked up with MTBE and water.
The organic phase was washed with water and brine. The solvent was
removed to afford
(2,2-difluoro-1,3-benzodioxol-5-yl)-cyclopropanecarbonitrile.
Preparation of
1-(2,2-difluoro-1,3-benzodioxol-5-yl)-cyclopropanecarboxylic
acid
##STR00026##
[0314] (2,2-difluoro-1,3-benzodioxol-5-yl)-cyclopropanecarbonitrile
was hydrolyzed using 6 M NaOH (8 equiv) in ethanol (5 vol) at
80.degree. C. overnight. The mixture was cooled to room temperature
and the ethanol was evaporated under vacuum. The residue was taken
up in water and MTBE, 1 M HCl was added, and the layers were
separated. The MTBE layer was then treated with dicyclohexylamine
(DCHA) (0.97 equiv). The slurry was cooled to 0.degree. C.,
filtered and washed with heptane to give the corresponding DCHA
salt. The salt was taken into MTBE and 10% citric acid and stirred
until all the solids had dissolved. The layers were separated and
the MTBE layer was washed with water and brine. A solvent swap to
heptane followed by filtration gave
1-(2,2-difluoro-1,3-benzodioxol-5-yl)-cyclopropanecarboxylic acid
after drying in a vacuum oven at 50.degree. C. overnight.
Preparation of
1-(2,2-difluoro-1,3-benzodioxol-5-yl)-cyclopropanecarbonyl
chloride
##STR00027##
[0316] 1-(2,2-difluoro-1,3-benzodioxol-5-yl)-cyclopropanecarboxylic
acid (1.2 eq) is slurried in toluene (2.5 vol) and the mixture was
heated to 60.degree. C. SOCl.sub.2 (1.4 eq) was added via addition
funnel. The toluene and SOCl.sub.2 were distilled from the reaction
mixture after 30 minutes. Additional toluene (2.5 vol) was added
and the resulting mixture was distilled again, leaving the product
acid chloride as an oil, which was used without further
purification.
Preparation of tert-butyl-3-(3-methylpyridin-2-yl)benzoate
##STR00028##
[0318] 2-Bromo-3-methylpyridine (1.0 eq) was dissolved in toluene
(12 vol). K.sub.2CO.sub.3 (4.8 eq) was added, followed by water
(3.5 vol). The resulting mixture was heated to 65.degree. C. under
a stream of N.sub.2 for 1 hour. 3-(t-Butoxycarbonyl)phenylboronic
acid (1.05 eq) and Pd(dppf)Cl.sub.2.CH.sub.2Cl.sub.2 (0.015 eq)
were then added and the mixture was heated to 80.degree. C. After 2
hours, the heat was turned off, water was added (3.5 vol), and the
layers were allowed to separate. The organic phase was then washed
with water (3.5 vol) and extracted with 10% aqueous methanesulfonic
acid (2 eq MsOH, 7.7 vol). The aqueous phase was made basic with
50% aqueous NaOH (2 eq) and extracted with EtOAc (8 vol). The
organic layer was concentrated to afford crude
tert-butyl-3-(3-methylpyridin-2-yl)benzoate (82%) that was used
directly in the next step.
Preparation of
2-(3-(tert-butoxycarbonyl)phenyl)-3-methylpyridine-1-oxide
##STR00029##
[0320] tert-Butyl-3-(3-methylpyridin-2-yl)benzoate (1.0 eq) was
dissolved in EtOAc (6 vol). Water (0.3 vol) was added, followed by
urea-hydrogen peroxide (3 eq). Phthalic anhydride (3 eq) was then
added portionwise to the mixture as a solid at a rate to maintain
the temperature in the reactor below 45.degree. C. After completion
of the phthalic anhydride addition, the mixture was heated to
45.degree. C. After stirring for an additional 4 hours, the heat
was turned off. 10% w/w aqueous Na.sub.2SO.sub.3 (1.5 eq) was added
via addition funnel. After completion of Na.sub.2SO.sub.3 addition,
the mixture was stirred for an additional 30 min and the layers
separated. The organic layer was stirred and 10% wt/wt aqueous.
Na.sub.2CO.sub.3 (2 eq) was added. After stirring for 30 minutes,
the layers were allowed to separate. The organic phase was washed
13% w/v aq NaCl. The organic phase was then filtered and
concentrated to afford crude
2-(3-(tert-butoxycarbonyl)phenyl)-3-methylpyridine-1-oxide (95%)
that was used directly in the next step.
Preparation of
tert-butyl-3-(6-amino-3-methylpyridin-2-yl)benzoate
##STR00030##
[0322] A solution of
2-(3-(tert-butoxycarbonyl)phenyl)-3-methylpyridine-1-oxide (1 eq)
and pyridine (4 eq) in acetonitrile (8 vol) was heated to
70.degree. C. A solution of methanesulfonic anhydride (1.5 eq) in
MeCN (2 vol) was added over 50 min via addition funnel while
maintaining the temperature at less than 75.degree. C. The mixture
was stirred for an additional 0.5 hours after complete addition.
The mixture was then allowed to cool to ambient. Ethanolamine (10
eq) was added via addition funnel. After stirring for 2 hours,
water (6 vol) was added and the mixture was cooled to 10.degree. C.
After stirring for 3 hours, the solid was collected by filtration
and washed with water (3 vol), 2:1 acetonitrile/water (3 vol), and
acetonitrile (2.times.1.5 vol). The solid was dried to constant
weight (<1% difference) in a vacuum oven at 50.degree. C. with a
slight N.sub.2 bleed to afford
tert-butyl-3-(6-amino-3-methylpyridin-2-yl)benzoate as a red-yellow
solid (53% yield).
Preparation of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-cyclopropanecarboxamido)-3-
-methylpyridin-2-yl)-t-butylbenzoate
##STR00031##
[0324] The crude acid chloride described above was dissolved in
toluene (2.5 vol based on acid chloride) and added via addition
funnel to a mixture of
tert-butyl-3-(6-amino-3-methylpyridin-2-yl)benzoate (1 eq), DMAP,
(0.02 eq), and triethylamine (3.0 eq) in toluene (4 vol based on
tert-butyl-3-(6-amino-3-methylpyridin-2-yl)benzoate). After 2
hours, water (4 vol based on
tert-butyl-3-(6-amino-3-methylpyridin-2-yl)benzoate) was added to
the reaction mixture. After stirring for 30 minutes, the layers
were separated. The organic phase was then filtered and
concentrated to afford a thick oil of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)-t-butylbenzoate (quantitative crude yield).
Acetonitrile (3 vol based on crude product) was added and distilled
until crystallization occurs. Water (2 vol based on crude product)
was added and the mixture stirred for 2 h. The solid was collected
by filtration, washed with 1:1 (by volume) acetonitrile/water
(2.times.1 volumes based on crude product), and partially dried on
the filter under vacuum. The solid was dried to a constant weight
(<1% difference) in a vacuum oven at 60.degree. C. with a slight
N.sub.2 bleed to afford
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)-t-butylbenzoate as a brown solid.
Preparation of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid.HCL salt
##STR00032##
[0326] To a slurry of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)-t-butylbenzoate (1.0 eq) in MeCN (3.0 vol) was
added water (0.83 vol) followed by concentrated aqueous HCl (0.83
vol). The mixture was heated to 45.+-.5.degree. C. After stirring
for 24 to 48 h, the reaction was complete, and the mixture was
allowed to cool to ambient. Water (1.33 vol) was added and the
mixture stirred. The solid was collected by filtration, washed with
water (2.times.0.3 vol), and partially dried on the filter under
vacuum. The solid was dried to a constant weight (<1%
difference) in a vacuum oven at 60.degree. C. with a slight N.sub.2
bleed to afford
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid.HCl as an off-white solid.
[0327] An .sup.1HNMR spectrum of Compound 1 is shown in FIG. 20 and
FIG. 21 depicts an .sup.1HNMR spectrum of Compound 1 as an HCl
salt.
[0328] Table 2 below recites the .sup.1HNMR data for Compound
I.
TABLE-US-00013 TABLE 2 Compound LC/MS LC/RT No M + 1 minutes NMR 1
453.3 1.93 .sup.1HNMR (400 MHz, DMSO-d6) 9.14 (s, 1H), 7.99-7.93
(m, 3H), 7.80-7.78 (m, 1H), 7.74-7.72 (m, 1H), 7.60-7.55 (m, 2H),
7.41-7.33 (m, 2H), 2.24 (s, 3H), 1.53-1.51 (m, 2H), 1.19-1.17 (m,
2H).
Preparation of Compound 1 Form I, Method A
##STR00033##
[0330] A slurry of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)benzoic acid.HCl (1 eq) in water (10 vol) was
stirred at ambient temperature. A sample was taken after stirring
for 24 h. The sample was filtered and the solid was washed with
water (2 times). The solid sample was submitted for DSC analysis.
When DSC analysis indicated complete conversion to Form I, the
solid was collected by filtration, washed with water (2.times.1.0
vol), and partially dried on a filter under vacuum. The solid was
then dried to a constant weight (<1% difference) in a vacuum
oven at 60.degree. C. with a slight N.sub.2 bleed to afford
Compound 1 Form I as an off-white solid (98% yield). .sup.1H NMR
(400 MHz, DMSO-d6) 9.14 (s, 1H), 7.99-7.93 (m, 3H), 7.80-7.78 (m,
1H), 7.74-7.72 (m, 1H), 7.60-7.55 (m, 2H), 7.41-7.33 (m, 2H), 2.24
(s, 3H), 1.53-1.51 (m, 2H), 1.19-1.17 (m, 2H).
Preparation of Compound 1 Form I, Method B
##STR00034##
[0332] A solution of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)-t-butylbenzoate (1.0 eq) in formic acid (3.0
vol) was heated with stirring to 70.+-.10.degree. C., for 8 h. The
reaction was deemed complete when no more than 1.0% AUC by
chromatographic methods of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)-t-butylbenzoate) remained. The mixture was
allowed to cool to ambient. The solution was added to water (6
vol), heated at 50.degree. C., and the mixture was stirred. The
mixture was then heated to 70.+-.10.degree. C. until the level of
3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3--
methylpyridin-2-yl)-t-butylbenzoate was no more than 0.8% (AUC).
The solid was collected by filtration, washed with water (2.times.3
vol), and partially dried on the filter under vacuum. The solid was
dried to a constant weight (<1% difference) in a vacuum oven at
60.degree. C. with a slight N.sub.2 bleed to afford Compound 1 Form
I as an off-white solid.
[0333] The DSC trace of Compound 1 Form I is shown in FIG. 22.
Melting for Compound 1 Form I occurs at about 204.degree. C.
[0334] An X-ray diffraction pattern was calculated from a single
crystal structure of Compound 1 Form I and is shown in FIG. 1.
Table 3 lists the calculated peaks for FIG. 1.
TABLE-US-00014 TABLE 3 2.theta. Angle Relative Peak Rank [degrees]
Intensity [%] 11 14.41 48.2 8 14.64 58.8 1 15.23 100.0 2 16.11 94.7
3 17.67 81.9 7 19.32 61.3 4 21.67 76.5 5 23.40 68.7 9 23.99 50.8 6
26.10 67.4 10 28.54 50.1
[0335] An actual X-ray powder diffraction pattern of Compound 1
Form I is shown in FIG. 2. Table 4 lists the actual peaks for FIG.
2.
TABLE-US-00015 TABLE 4 2.theta. Angle Relative Peak Rank [degrees]
Intensity [%] 7 7.83 37.7 3 14.51 74.9 4 14.78 73.5 1 15.39 100.0 2
16.26 75.6 6 16.62 42.6 5 17.81 70.9 9 21.59 36.6 10 23.32 34.8 11
24.93 26.4 8 25.99 36.9
[0336] Colorless crystals of Compound 1 Form I were obtained by
cooling a concentrated 1-butanol solution from 75.degree. C. to
10.degree. C. at a rate of 0.2.degree. C./min. A crystal with
dimensions of 0.50.times.0.08.times.0.03 mm was selected, cleaned
with mineral oil, mounted on a MicroMount and centered on a Bruker
APEX II system. Three batches of 40 frames separated in reciprocal
space were obtained to provide an orientation matrix and initial
cell parameters. Final cell parameters were obtained and refined
based on the full data set.
[0337] A diffraction data set of reciprocal space was obtained to a
resolution of 0.82 .ANG. using 0.5.degree. steps using 30 s
exposure for each frame. Data were collected at 100 (2) K.
Integration of intensities and refinement of cell parameters were
accomplished using APEXII software. Observation of the crystal
after data collection showed no signs of decomposition.
[0338] A conformational picture of Compound 1 Form I based on
single crystal X-ray analysis is shown in FIG. 23. Compound 1 Form
I is monoclinic, P.sub.21/n, with the following unit cell
dimensions: a=4.9626(7) .ANG., b=12.299(2) .ANG., c=33.075 (4)
.ANG., .beta.=93.938(9).degree., V=2014.0 .ANG..sup.3, Z=4. Density
of Compound 1 Form I calculated from structural data is 1.492
g/cm.sup.3 at 100 K.
Preparation of Compound 1 Form II from Compound 1 Form I
[0339] Compound 1 Form I (approximately 30 mg) was slurried in 500
.mu.L of an appropriate solvent (for example, methanol, ethanol,
acetone, 2-propanol, acetonitrile, tetrahydrofuran, methyl acetate,
2-butanone, ethyl formate, and -methyl tetrahydrofuran for two
days. The slurry was then filtered centrifugally or under vacuum
and was left to dry at ambient temperature overnight to yield
Compound 1 Form II.
[0340] The DSC trace of Compound 1 Form II Acetone Solvate is shown
in FIG. 15, showing two phase transitions. The melting point for
Compound 1 Form II Acetone Solvate occurs at about 188.degree. C.
and 205.degree. C.
[0341] An actual X-ray powder diffraction pattern of Compound 1
Form II is shown in FIG. 3. Table 5 lists the actual peaks for FIG.
3 in descending order of relative intensity.
TABLE-US-00016 TABLE 5 2.theta. Angle Relative Intensity [degrees]
[%] 21.70 100.0 8.98 65.5 11.04 57.4 18.16 55.9 23.06 55.4 20.63
53.1 22.22 50.2 18.57 49.1 16.66 47.2 19.86 35.0
[0342] Conformational depictions of Compound 1 Form II Acetone
Solvate based on single crystal X-ray analysis are shown in FIG.
24. The stoichiometry between Compound 1 Form II and acetone is
approximately 4.4:1 (4.48:1 calculated from .sup.1H NMR; 4.38:1
from X-ray). The crystal structure reveals a packing of the
molecules where there are two voids or pockets per unit cell, or 1
void per host molecule. In the acetone solvate, approximately 92
percent of voids are occupied by acetone molecules. Compound 1 Form
II is a monoclinic P2.sub.1/n space group with the following unit
cell dimensions: a=16.5235(10) .ANG., b=12.7425(8) .ANG., c=20.5512
(13) .ANG., .alpha.=90.degree., .beta.=103.736(4).degree.,
.gamma.=90.degree., V=4203.3(5) .ANG..sup.3,=4. The density of
Compound 1 in Compound 1 Form II calculated from structural data is
1.430/cm.sup.3 at 100 K.
[0343] A solid state .sup.13C NMR spectrum of Compound 1 Form II
Acetone Solvate is shown in FIG. 25. Table 6 provides chemical
shifts of the relevant peaks.
TABLE-US-00017 TABLE 6 Compound 1 Form II, Acetone Solvate .sup.13C
Chem. Shifts Peak # F1 [ppm] Intensity 1 202.8 6.05 2 173.3 62.66 3
171.9 20.53 4 153.5 28.41 5 150.9 21.68 6 150.1 19.49 7 143.2 45.74
8 142.3 42.68 9 140.1 37.16 10 136.6 26.82 11 135.9 30.1 12 134.6
39.39 13 133.2 23.18 14 131.0 60.92 15 128.5 84.58 16 116.0 34.64
17 114.2 23.85 18 112.4 25.3 19 110.9 24.12 20 107.8 18.21 21 32.0
54.41 22 22.2 20.78 23 18.8 100
[0344] A solid state .sup.19F NMR spectrum of Compound 1 Form II
Acetone Solvate is shown in FIG. 26. Peaks with an asterisk denote
spinning side bands. Table 7 provides chemical shifts of the
relevant peaks.
TABLE-US-00018 TABLE 7 Compound 1 Form II, Acetone Solvate .sup.19F
Chem. Shifts Peak # F1 [ppm] Intensity 1 -41.6 12.5 2 -46.4 6.77 3
-51.4 9.05
Preparation of Compound 1 HCl Salt Form A
[0345] Colorless crystals of Compound 1 HCl Salt Form A were
obtained by slow evaporation from a concentrated solution of the
HCl salt of Compound 1 in ethanol. A crystal with dimensions of
0.30.times.1/5.times.0.15 mm was selected, cleaned using mineral
oil, mounted on a MicroMount and centered on a Bruker APEXII
diffractometer. Three batches of 40 frames separated in reciprocal
space were obtained to provide an orientation matrix and initial
cell parameters. Final cell parameters were obtained and refined
based on the full data set.
[0346] FIG. 18 provides a conformational image of Compound 1 HCl
Salt Form A as a dimer, based on single crystal analysis. An X-ray
diffraction pattern of Compound 1 HCl Salt Form A calculated from
the crystal structure is shown in FIG. 27. Table 8 contains the
calculated peaks for FIG. 27 in descending order of relative
intensity.
TABLE-US-00019 TABLE 8 2.theta. Relative Intensity [degrees] [%]
8.96 100.00 17.51 48.20 18.45 34.60 10.33 32.10 16.01 18.90 11.94
18.40 8.14 16.20 10.10 13.90 16.55 13.30 9.54 10.10 16.55 13.30
Exemplary Oral Pharmaceutical Formulations Comprising Compound
1
[0347] A tablet was prepared with the components and amounts listed
in Table 9 for Exemplary Tablet 1A comprising 100 mg of API, i.e.
Compound 1 Form I. Exemplary Tablet 1A (formulated to have 100 mg
of Compound 1) is prepared using a dry roller compaction device
formulation process. In Table 9, grades/brands were
microcrystalline cellulose: Avicel PH102; mannitol: Pearlitol SD
100; croscarmellose sodium: Acdisol; and colloidal silica:
Cabosil.
TABLE-US-00020 TABLE 9 (% w/w) Roller Compaction Granule Blend
Compound 1 Form I 30 Microcrystalline cellulose 42.3 Mannitol 21.2
Croscarmellose Sodium 3 Sodium Lauryl Sulfate 1 Colloidal Silica
0.5 Magnesium Stearate 2 Tablet Composition (100 mg dose, 335 mg
image) Roller Compaction Granule Blend 99.5 Magnesium Stearate
0.5
[0348] A tablet was prepared with the components and amounts listed
in Table 10 for Exemplary Tablet 1B comprising 100 mg of API, i.e.
Compound 1 Form I. Exemplary Tablet 1B (formulated to have 100 mg
of Compound 1 Form I) is prepared using a wet high shear granule
formulation process. In Table 10, grades/brands were as follows.
High Shear Granule Blend--microcrystalline cellulose: Avicel PH101;
mannitol: Pearlitol C50; croscarmellose sodium: Acdisol;
polyvinylpyrrolidone: Kollidon PVP K30; and in the Tablet
Composition--croscarmellose sodium: Acdisol.
TABLE-US-00021 TABLE 10 (% w/w) High Shear Granule Blend Compound 1
Form I 50 Microcrystalline cellulose 30 Mannitol 13 Croscarmellose
Sodium 2 Polyvinylpyrrolidone 4 Sodium Lauryl Sulfate 1 Water
(removed during drying) 25-40% solids Tablet Composition (100 mg
dose, 205 mg image) High Shear Granule Blend 97.5 Croscarmellose
Sodium 2.0 Magnesium Stearate 0.5
[0349] A tablet was prepared with the components and amounts listed
in Table 11 for Exemplary Tablet 1C comprising 100 mg of API, i.e.
crystalline Compound 1 Form I. Exemplary Tablet 1C (formulated to
have 100 mg of crystalline Compound 1 Form I) is prepared using a
wet high shear granule formulation process. In Table 11,
grades/brands were as follows. High Shear Granule
Blend--microcrystalline cellulose: Avicel PH101; mannitol:
Pearlitol C50; croscarmellose sodium: Acdisol;
polyvinylpyrrolidone: Kollidon PVP K30; and in the Tablet
Composition--croscarmellose sodium: Acdisol.
TABLE-US-00022 TABLE 11 (% w/w) High Shear Granule Blend Compound 1
Form I 60 Microcrystalline cellulose 20 Mannitol 13 Croscarmellose
Sodium 2 Polyvinylpyrrolidone 4 Sodium Lauryl Sulfate 1 Water
(removed during drying) 25-40% solids Tablet Composition (100 mg
dose, 171 mg image) High Shear Granule Blend 97.5 Croscarmellose
Sodium 2.0 Magnesium Stearate 0.5
[0350] A tablet was prepared with the components and amounts listed
in Table 12 for Exemplary Tablet 1D comprising 200 mg of API, i.e.
crystalline Compound 1 Form I. Exemplary Tablet 1D (formulated to
have 200 mg of crystalline Compound 1 Form I) is prepared using a
wet high shear granule formulation process. In Table 12,
grades/brands were as follows. High Shear Granule
Blend--microcrystalline cellulose: Avicel PH101; mannitol:
Pearlitol C50; croscarmellose sodium: Acdisol;
polyvinylpyrrolidone: Kollidon PVP K30; and in the Tablet
Composition--microcrystalline cellulose: Avicel PH200;
croscarmellose sodium: Acdisol; and magnesium stearate: 5712.
TABLE-US-00023 TABLE 12 (% w/w) High Shear Granule Blend Compound 1
Form I 60 Microcrystalline cellulose 20 Mannitol 13 Croscarmellose
Sodium 2 Polyvinylpyrrolidone 4 Sodium Lauryl Sulfate 1 Water
(removed during drying) 25-40% solids Tablet Composition (200 mg
dose, 402 mg image) High Shear Granule Blend 83 Microcrystalline
cellulose 14 Croscarmellose Sodium 2 Magnesium Stearate 1.0
[0351] A tablet was prepared with the components and amounts listed
in Table 13 for Exemplary Tablet 1E comprising 200 mg of API, i.e.
crystalline Compound 1 Form I. Exemplary Tablet 1E (formulated to
have 200 mg of crystalline Compound 1 Form I) is prepared using a
wet high shear granule formulation process. In Table 13,
grades/brands were as follows. High Shear Granule
Blend--microcrystalline cellulose: Avicel PH101; mannitol:
Pearlitol C50; croscarmellose sodium: Acdisol;
polyvinylpyrrolidone: Kollidon PVP K30; and in the Core Tablet
Composition--microcrystalline cellulose: Avicel PH200;
croscarmellose sodium: Acdisol; and magnesium stearate: 5712; and
in the film coat--film coat: Opadry II; wax: Carnauba.
TABLE-US-00024 TABLE 13 mg High Shear Granule Blend Compound 1 Form
I 200 Microcrystalline cellulose 66 Mannitol 43 Croscarmellose
Sodium 7 Polyvinylpyrrolidone 13 Sodium Lauryl Sulfate 3 Core
Tablet Composition (200 mg dose, 400 mg image) High Shear Granule
Blend 332 Microcrystalline cellulose 56 Croscarmellose Sodium 8
Magnesium Stearate 4 Film Coated Tablet (200 mg dose, 412 mg image)
Core Tablet Composition 400 Film Coat 12 Wax 0.04
[0352] A tablet was prepared with the components and amounts listed
in Table 14 for Exemplary Tablet 1F comprising 200 mg of API, i.e.
crystalline Compound 1 Form I. Exemplary Tablet 1F (formulated to
have 200 mg of crystalline Compound 1 Form I) is prepared using a
wet high shear granule formulation process. In Table 14,
grades/brands were as follows. High Shear Granule
Blend--microcrystalline cellulose: Avicel PH101; mannitol:
Pearlitol C50; croscarmellose sodium: Acdisol;
polyvinylpyrrolidone: Kollidon PVP K30; and in the Core Tablet
Composition--microcrystalline cellulose: Avicel PH200;
croscarmellose sodium: Acdisol; and magnesium stearate: 5712; and
in the film coat--film coat: Opadry II; wax: Carnauba.
TABLE-US-00025 TABLE 14 mg High Shear Granule Blend Compound 1 Form
I 200 Microcrystalline cellulose 67 Mannitol 45 Croscarmellose
Sodium 7 Polyvinylpyrrolidone 10.4 Sodium Lauryl Sulfate 2.6 Core
Tablet Composition (200 mg dose, 400 mg image) High Shear Granule
Blend 332 Microcrystalline cellulose 56 Croscarmellose Sodium 8
Magnesium Stearate 4 Film Coated Tablet (200 mg dose, 412 mg image)
Core Tablet Composition 400 Film Coat 12 Wax 0.04
[0353] A tablet was prepared with the components and amounts listed
in Table 15 for Exemplary Tablet 1G comprising 100 mg of API, i.e.
crystalline Compound 1 Form I. Exemplary Tablet 1G (formulated to
have 100 mg of crystalline Compound 1 Form I) is prepared using a
wet high shear granule formulation process. In Table 15,
grades/brands were as follows. High Shear Granule
Blend--microcrystalline cellulose: Avicel PH101; mannitol:
Pearlitol C50; croscarmellose sodium: Acdisol;
polyvinylpyrrolidone: Kollidon PVP K30; and in the Tablet
Composition--croscarmellose sodium: Acdisol.
TABLE-US-00026 TABLE 15 (% w/w) High Shear Granule Blend Compound 1
Form I 70 Microcrystalline cellulose 12 Mannitol 11 Croscarmellose
Sodium 2 Polyvinylpyrrolidone 4 Sodium Lauryl Sulfate 1 Water
(removed during drying) 25-40% solids Tablet Composition (100 mg
dose, 147 mg image) High Shear Granule Blend 97.5 Croscarmellose
Sodium 2.0 Magnesium Stearate 0.5
[0354] A tablet was prepared with the components and amounts listed
in Table 16 for Exemplary Tablet 1H comprising 100 mg of API, i.e.
crystalline Compound 1 Form I or Form II. Exemplary Tablet 1H
(formulated to have 100 mg of crystalline Compound 1 Form I or Form
II) is prepared using a wet high shear granule formulation process.
In Table 16, grades/brands were as follows. High Shear Granule
Blend--microcrystalline cellulose: Avicel PH101; mannitol:
Pearlitol C50; croscarmellose sodium: Acdisol;
polyvinylpyrrolidone: Kollidon PVP K30; and in the Core Tablet
Composition--microcrystalline cellulose: Avicel PH200;
croscarmellose sodium: Acdisol; and magnesium stearate: 5712.
TABLE-US-00027 TABLE 16 (% w/w) High Shear Granule Blend Compound 1
Form I or Form II 61 Microcrystalline cellulose 20.3 Mannitol 13.2
Croscarmellose Sodium 2 Polyvinylpyrrolidone 2.7 Sodium Lauryl
Sulfate 0.7 Tablet Composition (100 mg dose, 197 mg image) High
Shear Granule Blend 83 Microcrystalline cellulose 14 Croscarmellose
Sodium 2 Magnesium Stearate 1
[0355] A tablet was prepared with the components and amounts listed
in Table 17 for Exemplary Tablet 11 comprising 100 mg of API, i.e.
crystalline Compound 1 Form I or Form II. Exemplary Tablet 11
(formulated to have 100 mg of crystalline Compound 1 Form I or Form
II) is prepared using a wet high shear granule formulation process.
In Table 17, grades/brands were as follows. High Shear Granule
Blend--microcrystalline cellulose: Avicel PH101; mannitol:
Pearlitol C50; croscarmellose sodium: Acdisol;
polyvinylpyrrolidone: Kollidon PVP K30; and in the Core Tablet
Composition--microcrystalline cellulose: Avicel PH200;
croscarmellose sodium: Acdisol; and magnesium stearate: 5712.
TABLE-US-00028 TABLE 17 mg High Shear Granule Blend Compound 1 Form
I or Form II 100 Microcrystalline cellulose 33.3 Mannitol 21.7
Croscarmellose Sodium 3.3 Polyvinylpyrrolidone 4.4 Sodium Lauryl
Sulfate 1.1 Core Tablet Composition (100 mg dose, 197 mg image)
High Shear Granule Blend 163.9 Microcrystalline cellulose 27.6
Croscarmellose Sodium 3.9 Magnesium Stearate 2.0
[0356] Tablet Formation from Roller Compaction Granule
Composition
[0357] Equipment/Process
[0358] Equipment
[0359] Roller Compactors: Alexanderwerk WP 120, Vector TF-Mini, or
Vector TF-Labo.
[0360] Screening/Weighing
[0361] Compound 1 and excipients may be screened prior to or after
weigh-out. Appropriate screen sizes are mesh 20, mesh 40, or mesh
60. Compound 1 may be pre-blended with one or more of the
excipients to simplify screening.
[0362] Blending
[0363] Compound 1 and excipients may be added to the blender in
different order. The blending may be performed in a Turbula blender
or a v-shell blender. The components may be blended for 10 minutes
without lubricant followed by additional blending with lubricant
for 3 minutes.
[0364] Roller Compaction
[0365] The blend may be roller compacted in ribbons and milled into
granules using an Alexanderwerk WP 120. The rolls used may be the
25 mm rolls using a compaction pressure of 18 to 50 bar, a roller
speed of 3 to 12 RPM, and a screw feeder speed of 20 to 80 RPM. The
screen sizes of the integrated mill may be 2 mm for the top screen
and 0.8 mm for the bottom screen.
[0366] Blending
[0367] The roller compacted granules may be blended with
extra-granular excipients such as fillers and lubricant using a
V-shell blender. The blending time may be 5, 3 or 1 minute(s).
[0368] Compression
[0369] The compression blend has been compressed into tablets using
a single station Riva MiniPress with 10 mm tooling. The weight of
the tablets for a 100 mg dose may be about 200, 250, or 300 mg.
[0370] Film Coating
[0371] Tablets may be film coated using a pan coater, such as, for
example an O'Hara Labcoat.
[0372] Printing
[0373] Film coated tablets may be printed with a monogram on one or
both tablet faces with, for example, a Hartnett Delta printer.
[0374] Tablet Formation from High Shear Granule Composition
[0375] Equipment/Process
[0376] Equipment
[0377] Granulator: Procept MiPro with a 250 ml or 1 L granulation
bowl.
[0378] Screening/Weighing
[0379] Compound 1 and excipients may be screened prior to or after
weigh-out. Possible screen sizes are mesh 20, mesh 40, or mesh 60.
Compound 1 may be pre-blended with one or more of the excipients to
simplify screening.
[0380] Granulation Operation
[0381] Granulation Fluid--SLS and binder are added to purified
water and mixed until dissolved. A suitable ratio is 2.5% w/w SLS
and 10.0% w/w PVP K30 in water.
[0382] Granulation--The excipients and compound 1 are added to the
granulation bowl. The order of addition may be Compound 1,
disintegrant, diluent, and filler. The components may be mixed in
the 250 ml bowl for 1 minute at impeller speed 1000 RPM and chopper
speed 1000 RPM. Granulation may be performed at an impeller speed
of 2000 RPM with a chopper speed of 4000 RPM while adding the
granulation fluid with a syringe pump at 1.5 to 4.5 g/min. The
fluid addition time may be 4 to 12 minutes. After the required
binder fluid is added, the granules may be wet-massed for about 10
seconds to about 1 minute. One notable advantage of the present
high shear granulation process is using a granulation fluid that
comprises both a surfactant and the binder for better granulation
through increased wettability. In one embodiment, the surfactant is
SLS.
[0383] Drying
[0384] The granules may be dried using a vacuum oven, tray dryer,
bi-conical dryer, or fluid bed drier. The granules have been dried
using a vacuum oven with a nitrogen purge.
[0385] Blending
[0386] The granules may be blended with extra-granular excipients.
The granules have been blended with extra-granular disintegrant,
diluent, filler, and lubricant. The granules have been blended
using the Turbula blender for 3 minutes pre-lubricant and 1 minute
with lubricant. A larger scale blender such as a 4-quart V-shell
blender may be used.
[0387] Compression
[0388] The compression blend has been compressed into tablets using
a single station Riva MiniPress with 8 mm, or 10 mm tooling. The
weight of the tablets for a 100 mg dose may be about 160, 200, or
250 mg.
[0389] Film Coating
[0390] Tablets may be film coated using a pan coater, such as, for
example an O'Hara Labcoat.
[0391] Printing
[0392] Film coated tablets may be printed with a monogram on one or
both tablet faces with, for example, a Hartnett Delta printer.
[0393] Dosing Administration Schedule
[0394] In another aspect, the invention relates to a method of
treating a CFTR mediated disease in a subject comprising
administering to a subject in need thereof an effective amount of
the pharmaceutical composition provided by the invention. In
another embodiment, the pharmaceutical composition is administered
to the subject once every two weeks. In another embodiment, the
pharmaceutical composition is administered to the subject once a
week. In another embodiment, the pharmaceutical composition is
administered to the subject once every three days. In another
embodiment, the pharmaceutical composition is administered to the
subject once a day. In one embodiment, when the pharmaceutical
composition is a tablet according to Table 9, 10, 11, 12, 13, 14,
15, 16, or 17 dosing is once a day.
Assays
[0395] Assays for Detecting and Measuring .DELTA.F508-CFTR
Correction Properties of Compounds
[0396] Membrane potential optical methods for assaying
.DELTA.F508-CFTR modulation properties of compounds.
[0397] The optical membrane potential assay utilized
voltage-sensitive FRET sensors described by Gonzalez and Tsien (See
Gonzalez, J. E. and R. Y. Tsien (1995) "Voltage sensing by
fluorescence resonance energy transfer in single cells" Biophys J
69(4): 1272-80, and Gonzalez, J. E. and R. Y. Tsien (1997)
"Improved indicators of cell membrane potential that use
fluorescence resonance energy transfer" Chem Biol 4(4): 269-77) in
combination with instrumentation for measuring fluorescence changes
such as the Voltage/Ion Probe Reader (VIPR) (See, Gonzalez, J. E.,
K. Oades, et al. (1999) "Cell-based assays and instrumentation for
screening ion-channel targets" Drug Discov Today 4(9):
431-439).
[0398] These voltage sensitive assays are based on the change in
fluorescence resonant energy transfer (FRET) between the
membrane-soluble, voltage-sensitive dye, DiSBAC.sub.2(3), and a
fluorescent phospholipid, CC2-DMPE, which is attached to the outer
leaflet of the plasma membrane and acts as a FRET donor. Changes in
membrane potential (V.sub.m) cause the negatively charged
DiSBAC.sub.2(3) to redistribute across the plasma membrane and the
amount of energy transfer from CC2-DMPE changes accordingly. The
changes in fluorescence emission were monitored using VIPR.TM. II,
which is an integrated liquid handler and fluorescent detector
designed to conduct cell-based screens in 96- or 384-well
microliter plates.
1. Identification of Correction Compounds
[0399] To identify small molecules that correct the trafficking
defect associated with .DELTA.F508-CFTR; a single-addition HTS
assay format was developed. The cells were incubated in serum-free
medium for 16 hrs at 37.degree. C. in the presence or absence
(negative control) of test compound. As a positive control, cells
plated in 384-well plates were incubated for 16 hrs at 27.degree.
C. to "temperature-correct" .DELTA.F508-CFTR. The cells were
subsequently rinsed 3.times. with Krebs Ringers solution and loaded
with the voltage-sensitive dyes. To activate .DELTA.F508-CFTR, 10
.mu.M forskolin and the CFTR potentiator, genistein (20 .mu.M),
were added along with Cl.sup.--free medium to each well. The
addition of Cl.sup.--free medium promoted Cl.sup.- efflux in
response to .DELTA.F508-CFTR activation and the resulting membrane
depolarization was optically monitored using the FRET-based
voltage-sensor dyes.
2. Identification of Potentiator Compounds
[0400] To identify potentiators of .DELTA.F508-CFTR, a
double-addition HTS assay format was developed. During the first
addition, a Cl.sup.--free medium with or without test compound was
added to each well. After 22 sec, a second addition of
Cl.sup.--free medium containing 2-10 .mu.M forskolin was added to
activate .DELTA.F508-CFTR. The extracellular Cl.sup.- concentration
following both additions was 28 mM, which promoted CF efflux in
response to .DELTA.F508-CFTR activation and the resulting membrane
depolarization was optically monitored using the FRET-based
voltage-sensor dyes.
3. Solutions
[0401] Bath Solution #1: (in mM) NaCl 160, KCl 4.5, CaCl.sub.2 2,
MgCl.sub.2 1, HEPES 10, pH 7.4 with NaOH.
[0402] Chloride-free bath solution: Chloride salts in Bath Solution
#1 are substituted with gluconate salts.
[0403] CC2-DMPE: Prepared as a 10 mM stock solution in DMSO and
stored at -20.degree. C. DiSBAC.sub.2(3): Prepared as a 10 mM stock
in DMSO and stored at -20.degree. C.
4. Cell Culture
[0404] NIH3T3 mouse fibroblasts stably expressing .DELTA.F508-CFTR
are used for optical measurements of membrane potential. The cells
are maintained at 37.degree. C. in 5% CO.sub.2 and 90% humidity in
Dulbecco's modified Eagle's medium supplemented with 2 mM
glutamine, 10% fetal bovine serum, 1.times.NEAA, .beta.-ME,
1.times. pen/strep, and 25 mM HEPES in 175 cm.sup.2 culture flasks.
For all optical assays, the cells were seeded at 30,000/well in
384-well matrigel-coated plates and cultured for 2 hrs at
37.degree. C. before culturing at 27.degree. C. for 24 hrs for the
potentiator assay. For the correction assays, the cells are
cultured at 27.degree. C. or 37.degree. C. with and without
compounds for 16-24 hours.
[0405] Electrophysiological Assays for Assaying 4F508-CFTR
Modulation Properties of Compounds
1. Ussing Chamber Assay
[0406] Using chamber experiments were performed on polarized
epithelial cells expressing .DELTA.F508-CFTR to further
characterize the .DELTA.F508-CFTR modulators identified in the
optical assays. FRT.sup..DELTA.F508-CFTR epithelial cells grown on
Costar Snapwell cell culture inserts were mounted in an Ussing
chamber (Physiologic Instruments, Inc., San Diego, Calif.), and the
monolayers were continuously short-circuited using a Voltage-clamp
System (Department of Bioengineering, University of Iowa, IA, and,
Physiologic Instruments, Inc., San Diego, Calif.). Transepithelial
resistance was measured by applying a 2-mV pulse. Under these
conditions, the FRT epithelia demonstrated resistances of 4
K.OMEGA./cm.sup.2 or more. The solutions were maintained at
27.degree. C. and bubbled with air. The electrode offset potential
and fluid resistance were corrected using a cell-free insert. Under
these conditions, the current reflects the flow of CF through
.DELTA.F508-CFTR expressed in the apical membrane. The I.sub.SC was
digitally acquired using an MP100A-CE interface and AcqKnowledge
software (v3.2.6; BIOPAC Systems, Santa Barbara, Calif.).
2. Identification of Correction Compounds
[0407] Typical protocol utilized a basolateral to apical membrane
Cl.sup.- concentration gradient. To set up this gradient, normal
ringer was used on the basolateral membrane, whereas apical NaCl
was replaced by equimolar sodium gluconate (titrated to pH 7.4 with
NaOH) to give a large Cl.sup.- concentration gradient across the
epithelium. All experiments were performed with intact monolayers.
To fully activate .DELTA.F508-CFTR, forskolin (10 .mu.M) and the
PDE inhibitor, IBMX (100 .mu.M), were applied followed by the
addition of the CFTR potentiator, genistein (50 .mu.M).
[0408] As observed in other cell types, incubation at low
temperatures of FRT cells stably expressing .DELTA.F508-CFTR
increases the functional density of CFTR in the plasma membrane. To
determine the activity of correction compounds, the cells were
incubated with 10 .mu.M of the test compound for 24 hours at
37.degree. C. and were subsequently washed 3.times. prior to
recording. The cAMP- and genistein-mediated I.sub.SC in
compound-treated cells was normalized to the 27.degree. C. and
37.degree. C. controls and expressed as percentage activity.
Preincubation of the cells with the correction compound
significantly increased the cAMP- and genistein-mediated I.sub.SC
compared to the 37.degree. C. controls.
3. Identification of Potentiator Compounds
[0409] Typical protocol utilized a basolateral to apical membrane
Cl.sup.- concentration gradient. To set up this gradient, normal
ringers was used on the basolateral membrane and was permeabilized
with nystatin (360 .mu.g/ml), whereas apical NaCl was replaced by
equimolar sodium gluconate (titrated to pH 7.4 with NaOH) to give a
large Cl.sup.- concentration gradient across the epithelium. All
experiments were performed 30 min after nystatin permeabilization.
Forskolin (10 .mu.M) and all test compounds were added to both
sides of the cell culture inserts. The efficacy of the putative
.DELTA.F508-CFTR potentiators was compared to that of the known
potentiator, genistein.
4. Solutions
[0410] Basolateral solution (in mM): NaCl (135), CaCl.sub.2 (1.2),
MgCl.sub.2 (1.2), K.sub.2HPO.sub.4 (2.4), KHPO.sub.4 (0.6),
N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) (10),
and dextrose (10). The solution was titrated to pH 7.4 with
NaOH.
[0411] Apical solution (in mM): Same as basolateral solution with
NaCl replaced with Na Gluconate (135).
5. Cell Culture
[0412] Fisher rat epithelial (FRT) cells expressing
.DELTA.F508-CFTR (FRT.sup..DELTA.F508-CFTR) were used for Ussing
chamber experiments for the putative .DELTA.F508-CFTR modulators
identified from our optical assays. The cells were cultured on
Costar Snapwell cell culture inserts and cultured for five days at
37.degree. C. and 5% CO.sub.2 in Coon's modified Ham's F-12 medium
supplemented with 5% fetal calf serum, 100 U/ml penicillin, and 100
.mu.g/ml streptomycin. Prior to use for characterizing the
potentiator activity of compounds, the cells were incubated at
27.degree. C. for 16-48 hrs to correct for the .DELTA.F508-CFTR. To
determine the activity of corrections compounds, the cells were
incubated at 27.degree. C. or 37.degree. C. with and without the
compounds for 24 hours.
6. Whole-Cell Recordings
[0413] The macroscopic .DELTA.F508-CFTR current (I.sub..DELTA.F508)
in temperature- and test compound-corrected NIH3T3 cells stably
expressing .DELTA.F508-CFTR were monitored using the
perforated-patch, whole-cell recording. Briefly, voltage-clamp
recordings of I.sub..DELTA.F508 were performed at room temperature
using an Axopatch 200B patch-clamp amplifier (Axon Instruments
Inc., Foster City, Calif.). All recordings were acquired at a
sampling frequency of 10 kHz and low-pass filtered at 1 kHz.
Pipettes had a resistance of 5-6 M.OMEGA. when filled with the
intracellular solution. Under these recording conditions, the
calculated reversal potential for Cl.sup.- (E.sub.Cl) at room
temperature was -28 mV. All recordings had a seal resistance>20
G.OMEGA. and a series resistance<15 M.OMEGA.. Pulse generation,
data acquisition, and analysis were performed using a PC equipped
with a Digidata 1320 A/D interface in conjunction with Clampex 8
(Axon Instruments Inc.). The bath contained <250 .mu.l of saline
and was continuously perifused at a rate of 2 ml/min using a
gravity-driven perfusion system,
7. Identification of Correction Compounds
[0414] To determine the activity of correction compounds for
increasing the density of functional .DELTA.F508-CFTR in the plasma
membrane, we used the above-described perforated-patch-recording
techniques to measure the current density following 24-hr treatment
with the correction compounds. To fully activate .DELTA.F508-CFTR,
10 .mu.M forskolin and 20 .mu.M genistein were added to the cells.
Under our recording conditions, the current density following 24-hr
incubation at 27.degree. C. was higher than that observed following
24-hr incubation at 37.degree. C. These results are consistent with
the known effects of low-temperature incubation on the density of
.DELTA.F508-CFTR in the plasma membrane. To determine the effects
of correction compounds on CFTR current density, the cells were
incubated with 10 .mu.M of the test compound for 24 hours at
37.degree. C. and the current density was compared to the
27.degree. C. and 37.degree. C. controls (% activity). Prior to
recording, the cells were washed 3.times. with extracellular
recording medium to remove any remaining test compound.
Preincubation with 10 .mu.M of correction compounds significantly
increased the cAMP- and genistein-dependent current compared to the
37.degree. C. controls.
8. Identification of Potentiator Compounds
[0415] The ability of .DELTA.F508-CFTR potentiators to increase the
macroscopic .DELTA.F508-CFTR Cl.sup.- current (I.sub..DELTA.F508)
in NIH3T3 cells stably expressing .DELTA.F508-CFTR was also
investigated using perforated-patch-recording techniques. The
potentiators identified from the optical assays evoked a
dose-dependent increase in I.sub..DELTA.F508 with similar potency
and efficacy observed in the optical assays. In all cells examined,
the reversal potential before and during potentiator application
was around -30 mV, which is the calculated E.sub.Cl (-28 mV).
9. Solutions
[0416] Intracellular solution (in mM): Cs-aspartate (90), CsCl
(50), MgCl.sub.2 (1), HEPES (10), and 240 .mu.g/ml amphotericin-B
(pH adjusted to 7.35 with CsOH).
[0417] Extracellular solution (in mM): N-methyl-D-glucamine
(NMDG)-Cl (150), MgCl.sub.2 (2), CaCl.sub.2 (2), HEPES (10) (pH
adjusted to 7.35 with HCl).
10. Cell Culture
[0418] NIH3T3 mouse fibroblasts stably expressing .DELTA.F508-CFTR
are used for whole-cell recordings. The cells are maintained at
37.degree. C. in 5% CO.sub.2 and 90% humidity in Dulbecco's
modified Eagle's medium supplemented with 2 mM glutamine, 10% fetal
bovine serum, 1.times.NEAA, .beta.-ME, 1.times. pen/strep, and 25
mM HEPES in 175 cm.sup.2 culture flasks. For whole-cell recordings,
2,500-5,000 cells were seeded on poly-L-lysine-coated glass
coverslips and cultured for 24-48 hrs at 27.degree. C. before use
to test the activity of potentiators; and incubated with or without
the correction compound at 37.degree. C. for measuring the activity
of correctors.
11. Single-Channel Recordings
[0419] The single-channel activities of temperature-corrected
.DELTA.F508-CFTR stably expressed in NIH3T3 cells and activities of
potentiator compounds were observed using excised inside-out
membrane patch. Briefly, voltage-clamp recordings of single-channel
activity were performed at room temperature with an Axopatch 200B
patch-clamp amplifier (Axon Instruments Inc.). All recordings were
acquired at a sampling frequency of 10 kHz and low-pass filtered at
400 Hz. Patch pipettes were fabricated from Corning Kovar Sealing
#7052 glass (World Precision Instruments, Inc., Sarasota, Fla.) and
had a resistance of 5-8 M.OMEGA. when filled with the extracellular
solution. The .DELTA.F508-CFTR was activated after excision, by
adding 1 mM Mg-ATP, and 75 nM of the cAMP-dependent protein kinase,
catalytic subunit (PKA; Promega Corp. Madison, Wis.). After channel
activity stabilized, the patch was perifused using a gravity-driven
microperfusion system. The inflow was placed adjacent to the patch,
resulting in complete solution exchange within 1-2 sec. To maintain
.DELTA.F508-CFTR activity during the rapid perifusion, the
nonspecific phosphatase inhibitor F (10 mM NaF) was added to the
bath solution. Under these recording conditions, channel activity
remained constant throughout the duration of the patch recording
(up to 60 min). Currents produced by positive charge moving from
the intra- to extracellular solutions (anions moving in the
opposite direction) are shown as positive currents. The pipette
potential (V.sub.p) was maintained at 80 mV.
[0420] Channel activity was analyzed from membrane patches
containing 2 active channels. The maximum number of simultaneous
openings determined the number of active channels during the course
of an experiment. To determine the single-channel current
amplitude, the data recorded from 120 sec of .DELTA.F508-CFTR
activity was filtered "off-line" at 100 Hz and then used to
construct all-point amplitude histograms that were fitted with
multigaussian functions using Bio-Patch Analysis software
(Bio-Logic Comp. France). The total microscopic current and open
probability (P.sub.o) were determined from 120 sec of channel
activity. The P.sub.o was determined using the Bio-Patch software
or from the relationship P.sub.o=I/i(N), where I=mean current,
i=single-channel current amplitude, and N=number of active channels
in patch.
12. Solutions
[0421] Extracellular solution (in mM): NMDG (150), aspartic acid
(150), CaCl.sub.2 (5), MgCl.sub.2 (2), and HEPES (10) (pH adjusted
to 7.35 with Tris base).
[0422] Intracellular solution (in mM): NMDG-Cl (150), MgCl.sub.2
(2), EGTA (5), TES (10), and Tris base (14) (pH adjusted to 7.35
with HCl).
13. Cell Culture
[0423] NIH3T3 mouse fibroblasts stably expressing .DELTA.F508-CFTR
are used for excised-membrane patch-clamp recordings. The cells are
maintained at 37.degree. C. in 5% CO.sub.2 and 90% humidity in
Dulbecco's modified Eagle's medium supplemented with 2 mM
glutamine, 10% fetal bovine serum, 1.times.NEAA, .beta.-ME,
1.times. pen/strep, and 25 mM HEPES in 175 cm.sup.2 culture flasks.
For single channel recordings, 2,500-5,000 cells were seeded on
poly-L-lysine-coated glass coverslips and cultured for 24-48 hrs at
27.degree. C. before use.
[0424] Using the procedures described above, the activity, i.e.,
EC50s, of Compound 1 has been measured and is shown in Table
18.
TABLE-US-00029 TABLE 18 IC50/EC50 Bins: +++ <= 2.0 < ++ <=
5.0 < + PercentActivity Bins: + <= 25.0 < ++ <= 100.0
< +++ Cmpd. No. BinnedEC50 BinnedMaxEfficacy 1 +++ +++
Other Embodiments
[0425] All publications and patents referred to in this disclosure
are incorporated herein by reference to the same extent as if each
individual publication or patent application were specifically and
individually indicated to be incorporated by reference. Should the
meaning of the terms in any of the patents or publications
incorporated by reference conflict with the meaning of the terms
used in this disclosure, the meaning of the terms in this
disclosure are intended to be controlling. Furthermore, the
foregoing discussion discloses and describes merely exemplary
embodiments of the invention. One skilled in the art will readily
recognize from such discussion and from the accompanying drawings
and claims, that various changes, modifications and variations can
be made therein without departing from the spirit and scope of the
invention as defined in the following claims.
* * * * *