U.S. patent application number 14/421454 was filed with the patent office on 2015-10-22 for compounds and methods for inhibiting nhe-mediated antiport in the treatment of disorders associated with fluid retention or salt overload and gastrointestinal tract disorders.
This patent application is currently assigned to ARDELYX, INC. The applicant listed for this patent is ARDELYX, INC.. Invention is credited to Noah BELL, Christopher CARRERAS, Dominique CHARMOT, Tao CHEN, Jeffrey JACOBS, Michael LEADBETTER, Jason LEWIS.
Application Number | 20150299131 14/421454 |
Document ID | / |
Family ID | 49036603 |
Filed Date | 2015-10-22 |
United States Patent
Application |
20150299131 |
Kind Code |
A1 |
BELL; Noah ; et al. |
October 22, 2015 |
COMPOUNDS AND METHODS FOR INHIBITING NHE-MEDIATED ANTIPORT IN THE
TREATMENT OF DISORDERS ASSOCIATED WITH FLUID RETENTION OR SALT
OVERLOAD AND GASTROINTESTINAL TRACT DISORDERS
Abstract
The present disclosure is directed to compounds and methods for
the treatment of disorders associated with fluid retention or salt
overload, such as heart failure (in particular, congestive heart
failure), chronic kidney disease, end-stage renal disease, liver
disease, and peroxisome proliferator-activated receptor (PPAR)
gamma agonist-induced fluid retention. The present disclosure is
also directed to compounds and methods for the treatment of
hypertension. The present disclosure is also directed to compounds
and methods for the treatment of gastrointestinal tract disorders,
including the treatment or reduction of pain associated with
gastrointestinal tract disorders.
Inventors: |
BELL; Noah; (Fremont,
CA) ; CARRERAS; Christopher; (Fremont, CA) ;
CHARMOT; Dominique; (Fremont, CA) ; CHEN; Tao;
(Fremont, CA) ; LEADBETTER; Michael; (Fremont,
CA) ; JACOBS; Jeffrey; (Fremont, CA) ; LEWIS;
Jason; (Fremont, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
ARDELYX, INC. |
Fremont |
CA |
US |
|
|
Assignee: |
ARDELYX, INC
Fremont
CA
|
Family ID: |
49036603 |
Appl. No.: |
14/421454 |
Filed: |
August 20, 2013 |
PCT Filed: |
August 20, 2013 |
PCT NO: |
PCT/GB2013/052193 |
371 Date: |
May 4, 2015 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
61691637 |
Aug 21, 2012 |
|
|
|
Current U.S.
Class: |
424/78.31 ;
424/606; 424/682; 424/692; 424/738; 514/18.3; 514/235.2;
514/253.05; 514/308; 514/53; 514/57; 544/128; 544/363; 546/140 |
Current CPC
Class: |
A61P 1/00 20180101; A61K
31/496 20130101; A61K 31/5377 20130101; A61K 45/06 20130101; C07D
217/18 20130101; A61P 9/12 20180101; C07D 401/14 20130101; A61K
47/60 20170801; A61P 1/10 20180101; A61P 31/12 20180101; A61P 13/12
20180101; A61K 31/4725 20130101; A61P 1/04 20180101; Y02A 50/30
20180101; A61P 43/00 20180101; A61K 47/36 20130101; C07D 217/14
20130101; A61K 47/38 20130101; A61K 47/55 20170801; A61K 47/32
20130101; A61P 7/10 20180101; A61P 9/04 20180101 |
International
Class: |
C07D 217/18 20060101
C07D217/18; A61K 45/06 20060101 A61K045/06; A61K 31/5377 20060101
A61K031/5377; A61K 47/38 20060101 A61K047/38; A61K 31/496 20060101
A61K031/496; A61K 47/32 20060101 A61K047/32; A61K 47/36 20060101
A61K047/36; A61K 31/4725 20060101 A61K031/4725; C07D 401/14
20060101 C07D401/14 |
Claims
1. A compound having the structure of Formula (I): ##STR00092## or
a stereoisomer, prodrug or pharmaceutically acceptable salt
thereof, wherein: (a) NHE is a NHE-inhibiting small molecule moiety
having the following structure of Formula (A): ##STR00093##
wherein: each R.sub.1, R.sub.2, R.sub.3, R.sub.5 and R.sub.9 are
independently selected from H, halogen, --NR.sub.7(CO)R.sub.8,
--(CO)NR.sub.7R.sub.8, --SO.sub.2--NR.sub.7R.sub.8,
--NR.sub.7SO.sub.2R.sub.8, --NR.sub.7R.sub.8, --OR.sub.7,
--SR.sub.7, --O(CO)NR.sub.7R.sub.8, --NR.sub.7(CO)OR.sub.8, and
--NR.sub.7SO.sub.2NR.sub.8, where R.sub.7 and R.sub.8 are
independently selected from H, C.sub.1-6alkyl, --C.sub.1-6alkyl-OH
or a bond linking the NHE-inhibiting small molecule to L, provided
at least one is a bond linking the NHE-inhibiting small molecule to
L; R.sub.4 is selected from H, C.sub.1-C.sub.7 alkyl, or a bond
linking the NHE-inhibiting small molecule to L; R.sub.6 is absent
or selected from H and C.sub.1-C.sub.7 alkyl; and Ar1 and Ar2
independently represent an aromatic ring or a heteroaromatic ring;
(b) Core is a Core moiety having the following structure of Formula
(B): ##STR00094## wherein: X is selected from C(X.sub.1), N and
N(C.sub.1-6alkyl); X.sub.1 is selected from hydrogen, optionally
substituted alkyl, --NX.sub.aN.sub.b, --NO.sub.2,
--NX.sub.c--C(.dbd.O)--NX.sub.c--X.sub.a,
--C(.dbd.O)NX.sub.c--X.sub.a, --NX.sub.c--C(.dbd.O)--X.sub.a,
--NX.sub.c--SO.sub.2--X.sub.a, --C(.dbd.O)--X.sub.a and --OX.sub.a,
each X.sub.a and X.sub.b are independently selected from hydrogen,
optionally substituted alkyl, optionally substituted cycloalkyl,
optionally substituted cycloalkylalkyl, optionally substituted
heterocyclyl, optionally substituted heterocyclylalkyl, optionally
substituted aryl, optionally substituted aralkyl, optionally
substituted heteroaryl and optionally substituted heteroarylalkyl;
Y is C.sub.1-6alkylene; Z is selected from
--NZ.sub.a--C(.dbd.O)--NZ.sub.a--, --C(.dbd.O)NZ.sub.a--,
--NZ.sub.a--C(.dbd.O)-- and heteroaryl when X is CX.sub.1; Z is
selected from --NZ.sub.a--C(.dbd.O)--NZ.sub.a--,
--NZ.sub.a--C(.dbd.O)-- and heteroaryl when X is N or
N(C.sub.1-6alkyl); and each X, and Z.sub.a is independently
selected from hydrogen and C.sub.1-6alkyl; and (c) L is a bond or
linker connecting the Core moiety to the NHE-inhibiting small
molecule moieties.
2. A compound of claim 1 wherein the NHE-inhibiting small molecule
moiety has the following structure: ##STR00095## wherein: each
R.sub.1, R.sub.2 and R.sub.3 are independently selected from H,
halogen, --NR.sub.7(CO)R.sub.8, --(CO)NR.sub.7R.sub.8,
--SO.sub.2--NR.sub.7R.sub.8, --NR.sub.7SO.sub.2R.sub.8,
--NR.sub.7R.sub.8, --OR.sub.7, --SR.sub.7, --O(CO)NR.sub.7R.sub.8,
--NR.sub.7(CO)OR.sub.8, and --NR.sub.7SO.sub.2NR.sub.8, where
R.sub.7 and R.sub.8 are independently selected from H,
C.sub.1-6alkyl, --C.sub.1-6alkyl-OH or a bond linking the
NHE-inhibiting small molecule to L, provided at least one is a bond
linking the NHE-inhibiting small molecule to L.
3. A compound of claim 2 wherein the NHE-inhibiting small molecule
moiety has one of the following structures: ##STR00096##
4. A compound of any of claims 1-3 wherein L is a polyalkylene
glycol linker.
5. A compound of any of claims 1-4 wherein L is a polyethylene
glycol linker.
6. A compound of any of claims 1-5 wherein X is C(X.sub.1).
7. A compound of claim 6 wherein each X, is hydrogen.
8. A compound of any of claims 1-5 wherein X is N.
9. A compound of any of claims 1-8 wherein each Z.sub.a is
hydrogen.
10. A compound having the structure of Formula (II): ##STR00097##
or a stereoisomer, prodrug or pharmaceutically acceptable salt
thereof, wherein: (a) NHE is a NHE-inhibiting small molecule moiety
having the structure of Formula (A): ##STR00098## wherein: each
R.sub.1, R.sub.2, R.sub.3, R.sub.5 and R.sub.9 are independently
selected from H, halogen, --NR.sub.7(CO)R.sub.8,
--(CO)NR.sub.7R.sub.8, --SO.sub.2--NR.sub.7R.sub.8,
--NR.sub.7SO.sub.2R.sub.8, --NR.sub.7R.sub.8, --OR.sub.7,
--SR.sub.7, --O(CO)NR.sub.7R.sub.8, --NR.sub.7(CO)OR.sub.8, and
--NR.sub.7SO.sub.2NR.sub.8, where R.sub.7 and R.sub.8 are
independently selected from H, C.sub.1-6alkyl, --C.sub.1-6alkyl-OH
or a bond linking the NHE-inhibiting small molecule to L, provided
at least one is a bond linking the NHE-inhibiting small molecule to
L; R.sub.4 is selected from H, C.sub.1-C.sub.7 alkyl, or a bond
linking the NHE-inhibiting small molecule to L; R.sub.6 is absent
or selected from H and C.sub.1-C.sub.7 alkyl; and Ar1 and Ar2
independently represent an aromatic ring or a heteroaromatic ring;
(b) Core is a Core moiety having the following structure of Formula
(C): ##STR00099## wherein: W is selected from alkylene,
polyalkylene glycol, --C(.dbd.O)--NH-(alkylene)-NH--C(.dbd.O)--,
--C(.dbd.O)--NH-(polyalkylene glycol)-NH--C(.dbd.O)--,
--C(.dbd.O)-(alkylene)-C(.dbd.O)--, --C(.dbd.O)-(polyalkylene
glycol)-C(.dbd.O)-- and cycloalkyl, X is N; Y is C.sub.1-6alkylene;
Z is selected from --NZ.sub.a--C(.dbd.O)--NZ.sub.a--,
--C(.dbd.O)NZ.sub.a--, --NZ.sub.a--C(.dbd.O)-- and heteroaryl; each
Z.sub.a is independently selected from hydrogen and C.sub.1-6alkyl;
and (c) L is a bond or linker connecting the Core moiety to the
NHE-inhibiting small molecules.
11. A compound of claim 10 wherein the NHE-inhibiting small
molecule moiety has the following structure: ##STR00100## wherein:
each R.sub.1, R.sub.2 and R.sub.3 are independently selected from
H, halogen, --NR.sub.7(CO)R.sub.8, --(CO)NR.sub.7R.sub.8,
--SO.sub.2--NR.sub.7R.sub.8, --NR.sub.7SO.sub.2R.sub.8,
--NR.sub.7R.sub.8, --OR.sub.7, --SR.sub.7, --O(CO)NR.sub.7R.sub.8,
--NR.sub.7(CO)OR.sub.8, and --NR.sub.7SO.sub.2NR.sub.8, where
R.sub.7 and R.sub.8 are independently selected from H,
C.sub.1-6alkyl, --C.sub.1-6alkyl-OH or a bond linking the
NHE-inhibiting small molecule to L, provided at least one is a bond
linking the NHE-inhibiting small molecule to L.
12. A compound of claim 11 wherein the NHE-inhibiting small
molecule moiety has one of the following structures:
##STR00101##
13. A compound of any of claims 10-12 wherein L is a polyalkylene
glycol linker.
14. A compound of any of claims 10-13 wherein L is a polyethylene
glycol linker.
15. A pharmaceutical composition comprising a compound of any of
claims 1-14, or a stereoisomer, pharmaceutically acceptable salt or
prodrug thereof, and a pharmaceutically acceptable carrier, diluent
or excipient.
16. A pharmaceutical composition of claim 15, further comprising a
fluid-absorbing polymer.
17. A pharmaceutical composition of claim 16 wherein the
fluid-absorbing polymer is delivered directly to the colon.
18. A pharmaceutical composition of claim 16 or 17 wherein the
fluid-absorbing polymer has a fluid absorbency of at least about 15
g of isotonic fluid per g of polymer under a static pressure of
about 5 kPa.
19. A pharmaceutical composition of any of claims 16-18 wherein the
fluid-absorbing polymer has a fluid absorbency of at least about 15
g of isotonic fluid per g of polymer under a static pressure of
about 10 kPa.
20. A pharmaceutical composition of any of claims 16-19 wherein the
fluid-absorbing polymer is characterized by a fluid absorbency of
at least about 10 g/g.
21. A pharmaceutical composition of any of claims 16-20 wherein the
fluid-absorbing polymer is characterized by a fluid absorbency of
at least about 15 g/g.
22. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is superabsorbent.
23. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is a crosslinked, partially neutralized
polyelectrolyte hydrogel.
24. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is a crosslinked polyacrylate.
25. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is a polyelectrolyte.
26. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is calcium Carbophil.
27. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is prepared by a high internal phase
emulsion process.
28. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is a foam.
29. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is prepared by a aqueous free radical
polymerization of acrylamide or a derivative thereof, a crosslinker
and a free radical initiator redox system in water.
30. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is a hydrogel.
31. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is an N-alkyl acrylamide.
32. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is a superporous gel.
33. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is naturally occurring.
34. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is selected from the group consisting of
xanthan, guar, wellan, hemicelluloses, alkyl-cellulose
hydro-alkyl-cellulose, carboxy-alkyl-cellulose, carrageenan,
dextran, hyaluronic acid and agarose.
35. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is psyllium.
36. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is a polysaccharide that includes xylose
and arabinose.
37. A pharmaceutical composition of any of claims 16-21 wherein the
fluid-absorbing polymer is a polysaccharide that includes xylose
and arabinose, wherein the ratio of xylose to arabinose is at least
about 3:1, by weight.
38. A pharmaceutical composition of any of claims 16-37, further
comprising another pharmaceutically active agent or compound.
39. A pharmaceutical composition of claim 38 wherein the
composition further comprises another pharmaceutically active agent
or compound selected from the group consisting of a diuretic,
cardiac glycoside, ACE inhibitor, angiotensin-2 receptor
antagonist, aldosterone antagonist, aldosterone synthase inhibitor,
renin inhibitor, calcium channel blocker, beta blocker, alpha
blocker, central alpha agonist, vasodilator, blood thinner,
anti-platelet agent, lipid-lowering agent, and peroxisome
proliferator-activated receptor (PPAR) gamma agonist agent.
40. A pharmaceutical composition of claim 39 wherein the diuretic
is selected from the group consisting of a high ceiling loop
diuretic, a benzothiadiazide diuretic, a potassium sparing
diuretic, and a osmotic diuretic.
41. A pharmaceutical composition of claim 38 wherein the
composition further comprises another pharmaceutically active agent
or compound selected from the group consisting of an analgesic
peptide or agent.
42. A pharmaceutical composition of claim 41 wherein the
composition further comprises another pharmaceutically active agent
or compound selected from the group consisting of a laxative agent
selected from a bulk-producing agent (e.g. psyllium husk
(Metamucil)), methylcellulose (Citrucel), polycarbophil, dietary
fiber, apples, stool softeners/surfactant (e.g., docusate, Colace,
Diocto), a hydrating or osmotic agent (e.g., dibasic sodium
phosphate, magnesium citrate, magnesium hydroxide (Milk of
magnesia), magnesium sulfate (which is Epsom salt), monobasic
sodium phosphate, sodium biphosphate), a hyperosmotic agent (e.g.,
glycerin suppositories, sorbitol, lactulose, and polyethylene
glycol (PEG)).
43. A method for inhibiting NHE-mediated antiport of sodium and
hydrogen ions, the method comprising administering to a mammal in
need thereof a pharmaceutically effective amount of a compound or
pharmaceutical composition of any of claims 1-42.
44. A method for treating a disorder associated with fluid
retention or salt overload, the method comprising administering to
a mammal in need thereof a pharmaceutically effective amount of a
compound or pharmaceutical composition of any of claims 1-42.
45. A method for treating a disorder selected from the group
consisting of heart failure, chronic kidney disease, end-stage
renal disease, liver disease, and peroxisome proliferator-activated
receptor (PPAR) gamma agonist-induced fluid retention, the method
comprising administering to a mammal in need thereof a
pharmaceutically effective amount of a compound or pharmaceutical
composition of any of claims 1-42.
46. The method of claim 45 wherein the heart failure is congestive
heart failure.
47. A method for treating hypertension, the method comprising
administering to a mammal in need thereof a pharmaceutically
effective amount of a compound or pharmaceutical composition of any
of claims 1-42.
48. A method of any of claims 44-47 wherein the method comprises
administering a pharmaceutically effective amount of the compound
to the mammal in order to increase the mammal's daily fecal output
of sodium and/or fluid.
49. A method of any of claims 44-48 wherein the method comprises
administering a pharmaceutically effective amount of the compound
to the mammal in order to increase the mammal's daily fecal output
of sodium by at least about 30 mmol, and/or fluid by at least about
200 ml.
50. A method of any of claims 44-49 wherein the mammal's fecal
output of sodium and/or fluid is increased without introducing
another type of cation in a stoichiometric or near stoichiometric
fashion via an ion exchange process.
51. A method of any of claims 44-50, further comprising
administering to the mammal a fluid-absorbing polymer to absorb
fecal fluid resulting from the use of the compound that is
substantially active in the gastrointestinal tract to inhibit
NHE-mediated antiport of sodium ions and hydrogen ions therein.
52. A method of any of claims 44-51 wherein the compound or
composition is administered to treat hypertension.
53. A method of any of claims 44-52 wherein the compound or
composition is administered to treat hypertension associated with
dietary salt intake.
54. A method of any of claims 44-51 wherein administration of the
compound or composition allows the mammal to intake a more
palatable diet.
55. A method of any of claims 44-51 wherein the compound or
composition is administered to treat fluid overload.
56. A method of claim 55 wherein the fluid overload is associated
with congestive heart failure.
57. A method of claim 55 wherein the fluid overload is associated
with end stage renal disease.
58. A method of claim 55 wherein the fluid overload is associated
with peroxisome proliferator-activated receptor (PPAR) gamma
agonist therapy.
59. A method of any of claims 44-51 wherein the compound or
composition is administered to treat sodium overload.
60. A method of any of claim 44-51 wherein the compound or
composition is administered to reduce interdialytic weight gain in
ESRD patients.
61. A method of any of claims 44-51 wherein the compound or
composition is administered to treat edema.
62. A method of claim 61 wherein the edema is caused by
chemotherapy, pre-menstrual fluid overload or preeclampsia.
63. A method of any of claims 44-62 wherein the compound or
composition is administered orally, by rectal suppository, or
enema.
64. A method of any one of claims 44-63, wherein the method
comprises administering a pharmaceutically effective amount of the
compound or composition in combination with one or more additional
pharmaceutically active compounds or agents.
65. A method of claim 64 wherein the one or more additional
pharmaceutically active compounds or agents is selected from the
group consisting of a diuretic, cardiac glycoside, ACE inhibitor,
angiotensin-2 receptor antagonist, aldosterone antagonist,
aldosterone synthase inhibitor, renin inhibitor, calcium channel
blocker, beta blocker, alpha blocker, central alpha agonist,
vasodilator, blood thinner, anti-platelet agent, lipid-lowering
agent, and peroxisome proliferator-activated receptor (PPAR) gamma
agonist agent.
66. A method of claim 65 wherein the diuretic is selected from the
group consisting of a high ceiling loop diuretic, a
benzothiadiazide diuretic, a potassium sparing diuretic, and a
osmotic diuretic.
67. A method of any of claims 64-66 wherein the pharmaceutically
effective amount of the compound or composition, and the one or
more additional pharmaceutically active compounds or agents, are
administered as part of a single pharmaceutical preparation.
68. A method of any of claims 64-66 wherein the pharmaceutically
effective amount of the compound or composition, and the one or
more additional pharmaceutically active compounds or agents, are
administered as individual pharmaceutical preparations.
69. A method of claim 68 wherein the individual pharmaceutical
preparation are administered sequentially.
70. A method of claim 69 wherein the individual pharmaceutical
preparation are administered simultaneously.
71. A method for treating a gastrointestinal tract disorder, the
method comprising administering to a mammal in need thereof a
pharmaceutically effective amount of a compound or pharmaceutical
composition of any of claims 1-42.
72. A method of claim 71 wherein the gastrointestinal tract
disorder is a gastrointestinal motility disorder.
73. A method of claim 71 wherein the gastrointestinal tract
disorder is irritable bowel syndrome.
74. A method of claim 71 wherein the gastrointestinal tract
disorder is chronic constipation.
75. A method of claim 71 wherein the gastrointestinal tract
disorder is chronic idiopathic constipation.
76. A method of claim 71 wherein the gastrointestinal tract
disorder is chronic constipation occurring in cystic fibrosis
patients.
77. A method of claim 71 wherein the gastrointestinal tract
disorder is opioid-induced constipation.
78. A method of claim 71 wherein the gastrointestinal tract
disorder is a functional gastrointestinal tract disorder.
79. A method of claim 71 wherein the gastrointestinal tract
disorder is selected from the group consisting of chronic
intestinal pseudo-obstruction and colonic pseudo-obstruction.
80. A method of claim 71 wherein the gastrointestinal tract
disorder is Crohn's disease.
81. A method of claim 71 wherein the gastrointestinal tract
disorder is ulcerative colitis.
82. A method of claim 71 wherein the gastrointestinal tract
disorder is a disease referred to as inflammatory bowel
disease.
83. A method of claim 71 wherein the gastrointestinal tract
disorder is associated with chronic kidney disease (stage 4 or
5).
84. A method of claim 71 wherein the gastrointestinal tract
disorder is constipation induced by calcium supplement.
85. A method of claim 71 wherein the gastrointestinal tract
disorder is constipation, and further wherein the constipation to
be treated is associated with the use of a therapeutic agent.
86. A method of claim 71 wherein the gastrointestinal tract
disorder is constipation, and further wherein the constipation to
be treated is associated with a neuropathic disorder.
87. A method of claim 71 wherein the gastrointestinal tract
disorder is constipation, and further wherein the constipation to
be treated is post-surgical constipation (postoperative ileus).
88. A method of claim 71 wherein the gastrointestinal tract
disorder is constipation, and further wherein the constipation to
be treated is idiopathic (functional constipation or slow transit
constipation).
89. A method of claim 71 wherein the gastrointestinal tract
disorder is constipation, and further wherein the constipation to
be treated is associated with neuropathic, metabolic or an
endocrine disorder (e.g., diabetes mellitus, renal failure,
hypothyroidism, hyperthyroidism, hypocalcaemia, Multiple Sclerosis,
Parkinson's disease, spinal cord lesions, neurofibromatosis,
autonomic neuropathy, Chagas disease, Hirschsprung's disease or
cystic fibrosis, and the like).
90. A method of claim 71 wherein the gastrointestinal tract
disorder is constipation, and further wherein the constipation to
be treated is due the use of drugs selected from analgesics (e.g.,
opioids), antihypertensives, anticonvulsants, antidepressants,
antispasmodics and antipsychotics.
91. A method for treating irritable bowel syndrome, the method
comprising administering to a mammal in need thereof a
pharmaceutically effective amount of a compound or a pharmaceutical
composition of any of claims 1-42.
92. A method of any of claims 71-91 wherein the compound or
composition is administered to treat or reduce pain associated with
a gastrointestinal tract disorder.
93. A method of any of claims 71-91 wherein the compound or
composition is administered to treat or reduce visceral
hypersensitivity associated with a gastrointestinal tract
disorder.
94. A method of any of claims 71-91 wherein the compound or
composition is administered to treat or reduce inflammation of the
gastrointestinal tract.
95. A method of any of claims 71-91 wherein the compound or
composition is administered to reduce gastrointestinal transit
time.
96. A method of any of claims 71-95 wherein the compound or
composition is administered either orally or by rectal
suppository.
97. A method of any of claims 71-96 wherein the method comprises
administering a pharmaceutically effective amount of the compound
or composition, in combination with one or more additional
pharmaceutically active compounds or agents.
98. A method of claim 97 wherein the one or more additional
pharmaceutically active agents or compounds are an analgesic
peptide or agent.
99. A method of claim 97 wherein the one or more additional
pharmaceutically active agents or compounds are selected from the
group consisting of a laxative agent selected from a bulk-producing
agent (e.g. psyllium husk (Metamucil)), methylcellulose (Citrucel),
polycarbophil, dietary fiber, apples, stool softeners/surfactant
(e.g., docusate, Colace, Diocto), a hydrating or osmotic agent
(e.g., dibasic sodium phosphate, magnesium citrate, magnesium
hydroxide (Milk of magnesia), magnesium sulfate (which is Epsom
salt), monobasic sodium phosphate, sodium biphosphate), and a
hyperosmotic agent (e.g., glycerin suppositories, sorbitol,
lactulose, and polyethylene glycol (PEG)).
100. A method of any of claims 97-99 wherein the pharmaceutically
effective amount of the compound or composition, and the one or
more additional pharmaceutically active compounds or agents, are
administered as part of a single pharmaceutical preparation.
101. A method of any of claims 97-99 wherein the pharmaceutically
effective amount of the compound or composition, and the one or
more additional pharmaceutically active compounds or agents, are
administered as individual pharmaceutical preparations.
102. A method of claim 101 wherein the individual pharmaceutical
preparation are administered sequentially.
103. A method of claim 101 wherein the individual pharmaceutical
preparation are administered simultaneously.
Description
RELATED APPLICATIONS
[0001] This application is a. National Stage application under 35
U.S.C. .sctn.371 of International Application No.
PCT/GB2013/052193, filed Aug. 20, 2013, which claims the benefit of
priority to U.S. Provisional Patent Application No. 61/691,637,
filed Aug. 21, 2012. The contents of the foregoing applications are
hereby incorporated by reference in their entirety.
BACKGROUND
[0002] 1. Field
[0003] The present disclosure is directed to compounds that are
substantially active in the gastrointestinal tract to inhibit
NHE-mediated antiport of sodium ions and hydrogen ions, and the use
of such compounds in the treatment of disorders associated with
fluid retention or salt overload and in the treatment of
gastrointestinal tract disorders, including the treatment or
reduction of pain associated with a gastrointestinal tract
disorder.
[0004] 2. Description of the Related Art
[0005] Disorders Associated with Fluid Retention and Salt
Overload
[0006] According to the American Heart Association, more than 5
million Americans have suffered from heart failure, and an
estimated 550,000 cases of congestive heart failure (CHF) occur
each year (Schocken, D. D. et al., Prevention of heart failure: a
scientific statement from the American Heart Association Councils
on Epidemiology and Prevention, Clinical Cardiology, Cardiovascular
Nursing, and High Blood Pressure Research; Quality of Care and
Outcomes Research Interdisciplinary Working Group; and Functional
Genomics and Translational Biology Interdisciplinary Working Group:
Circulation, v. 117, no. 19, p. 2544-2565 (2008)). The clinical
syndrome of congestive heart failure occurs when cardiac
dysfunction prevents adequate perfusion of peripheral tissues. The
most common form of heart failure leading to CHF is systolic heart
failure, caused by contractile failure of the myocardium. A main
cause of CHF is due to ischemic coronary artery disease, with or
without infarction. Long standing hypertension, particularly when
it is poorly controlled, may lead to CHF.
[0007] In patients with CHF, neurohumoral compensatory mechanisms
(i.e., the sympathetic nervous system and the renin-angiotensin
system) are activated in an effort to maintain normal circulation.
The renin-angiotensin system is activated in response to decreased
cardiac output, causing increased levels of plasma renin,
angiotensin II, and aldosterone. As blood volume increases in the
heart, cardiac output increases proportionally, to a point where
the heart is unable to dilate further. In the failing heart,
contractility is reduced, so the heart operates at higher volumes
and higher filling pressures to maintain output. Filling pressures
may eventually increase to a level that causes transudation of
fluid into the lungs and congestive symptoms (e.g., edema,
shortness of breath). All of these symptoms are related to fluid
volume and salt retention, and this chronic fluid and salt overload
further contribute to disease progression.
[0008] Compliance with the medication regimen and with dietary
sodium restrictions is a critical component of self-management for
patients with heart failure and may lengthen life, reduce
hospitalizations and improve quality of life. Physicians often
recommend keeping salt intake below 2.3 g per day and no more than
2 g per day for people with heart failure. Most people eat
considerably more than this, so it is likely that a person with
congestive heart failure will need to find ways to reduce dietary
salt.
[0009] A number of drug therapies currently exist for patients
suffering from CHF. For example, diuretics may be used or
administered to relieve congestion by decreasing volume and,
consequently, filling pressures to below those that cause pulmonary
edema. By counteracting the volume increase, diuretics reduce
cardiac output; however, fatigue and dizziness may replace CHF
symptoms. Among the classes or types of diuretics currently being
used is thiazides. Thiazides inhibit NaCl transport in the kidney,
thereby preventing reabsorption of Na in the cortical diluting
segment at the ending portion of the loop of Henle and the proximal
portion of the distal convoluted tubule. However, these drugs are
not effective when the glomerular filtration rate (GFR) is less
than 30 ml/min. Additionally, thiazides, as well as other
diuretics, may cause hypokalemia. Also among the classes or types
of diuretics currently being used is loop diuretics (e.g.,
furosemide). These are the most potent diuretics and are
particularly effective in treating pulmonary edema. Loop diuretics
inhibit the NaKCl transport system, thus preventing reabsorption of
Na in the loop of Henle.
[0010] Patients that have persistent edema despite receiving high
doses of diuretics may be or become diuretic-resistant. Diuretic
resistance may be caused by poor availability of the drug. In
patients with renal failure, which has a high occurrence in the CHF
population, endogenous acids compete with loop diuretics such as
furosemide for the organic acid secretory pathway in the tubular
lumen of the nephron. Higher doses, or continuous infusion, are
therefore needed to achieve entrance of an adequate amount of drug
into the nephron. However, recent meta-analysis have raised
awareness about the long-term risk of chronic use of diuretics in
the treatment of CHF. For instance, in a recent study (Ahmed et
al., Int J Cardiol. 2008 April 10; 125(2): 246-253) it was shown
that chronic diuretic use was associated with significantly
increased mortality and hospitalization in ambulatory older adults
with heart failure receiving angiotensin converting enzyme
inhibitor and diuretics.
[0011] Angiotensin-converting enzyme ("ACE") inhibitors are an
example of another drug therapy that may be used to treat
congestive heart failure. ACE inhibitors cause vasodilatation by
blocking the renin-angiotensin-aldosterone system. Abnormally low
cardiac output may cause the renal system to respond by releasing
renin, which then converts angiotensinogen into angiotensin I. ACE
converts angiotensin I into angiotensin II. Angiotensin II
stimulates the thirst centers in the hypothalamus and causes
vasoconstriction, thus increasing blood pressure and venous return.
Angiotensin II also causes aldosterone to be released, causing
reabsorption of Na and concomitant passive reabsorption of fluid,
which in turn causes the blood volume to increase. ACE inhibitors
block this compensatory system and improve cardiac performance by
decreasing systemic and pulmonary vascular resistance. ACE
inhibitors have shown survival benefit and conventionally have been
a treatment of choice for CHF. However, since ACE inhibitors lower
aldosterone, the K-secreting hormone, one of the side-effects of
their use is hyperkalemia. In addition, ACE inhibitors have been
show to lead to acute renal failure in certain categories of CHF
patients. (See, e.g., C. S. Cruz et al., "Incidence and Predictors
of Development of Acute Renal Failure Related to the Treatment of
Congestive Heart Failure with ACE Inhibitors, Nephron Clin. Pract.,
v. 105, no. 2, pp c77-c83 (2007)).
[0012] Patients with end stage renal disease ("ESRD"), i.e., stage
5 chronic kidney failure, must undergo hemodialysis three times per
week. The quasi-absence of renal function and ability to eliminate
salt and fluid results in large fluctuations in body weight as
fluid and salt build up in the body (sodium/volume overload). The
fluid overload is characterized as interdialytic weight gain. High
fluid overload is also worsened by heart dysfunction, specifically
CHF. Dialysis is used to remove uremic toxins and also adjust salt
and fluid homeostasis. However, symptomatic intradialytic
hypotension (SIH) may occur when patients are over-dialyzed. SIH is
exhibited in about 15% to 25% of the ESRD population (Davenport,
A., C. Cox, and R. Thuraisingham, Blood pressure control and
symptomatic intradialytic hypotension in diabetic haemodialysis
patients: a cross-sectional survey; Nephron Clin. Pract., v. 109,
no. 2, p. c65-c71 (2008)). Like in hypertensive and CHF patients,
dietary restrictions of salt and fluid are highly recommended but
poorly followed because of the poor palatability of low-salt
food
[0013] The cause of primary or "essential" hypertension is elusive.
However, several observations point to the kidney as a primary
factor. The strongest data for excess salt intake and elevated
blood pressure come from INTERSALT, a cross-sectional study of
greater than 10,000 participants. For individuals, a significant,
positive, independent linear relation between 24-hour sodium
excretion and systolic blood pressure was found. Higher individual
24-hour urinary sodium excretions were found to be associated with
higher systolic/diastolic blood pressure on average, by 6-3/3-0 mm
Hg. Primary hypertension is a typical example of a complex,
multifactorial, and polygenic trait. All these monogenic
hypertensive syndromes are virtually confined to mutated genes
involving gain of function of various components of the
renin-angiotensin-aldosterone system, resulting in excessive renal
sodium retention. In a broad sense, these syndromes are
characterized by increased renal sodium reabsorption arising
through either primary defects in sodium transport systems or
stimulation of mineralocorticoid receptor activity (Altun, B., and
M. Arici, 2006, Salt and blood pressure: time to challenge;
Cardiology, v. 105, no. 1, p. 9-16 (2006)). A much larger number of
controlled studies have been performed on hypertensive subjects
during the last three decades to determine whether sodium reduction
will reduce established high blood pressure. Meta-analyses of these
studies have clearly shown a large decrease in blood pressure in
hypertensive patients.
[0014] In end stage liver disease (ESLD), accumulation of fluid as
ascites, edema or pleural effusion due to cirrhosis is common and
results from a derangement in the extracellular fluid volume
regulatory mechanisms. Fluid retention is the most frequent
complication of ESLD and occurs in about 50% of patients within 10
years of the diagnosis of cirrhosis. This complication
significantly impairs the quality of life of cirrhotic patients and
is also associated with poor prognosis. The one-year and five-year
survival rate is 85% and 56%, respectively (Kashani et al., Fluid
retention in cirrhosis: pathophysiology and management; QJM, v.
101, no. 2, p. 71-85 (2008)). The most acceptable theories
postulate that the initial event in ascites formation in the
cirrhotic patient is sinusoidal hypertension. Portal hypertension
due to an increase in sinusoidal pressure activates vasodilatory
mechanisms. In advanced stages of cirrhosis, arteriolar
vasodilation causes underfilling of systemic arterial vascular
space. This event, through a decrease in effective blood volume,
leads to a drop in arterial pressure. Consequently,
baroreceptor-mediated activation of renin-angiotensin aldosterone
system, sympathetic nervous system and nonosmotic release of
antidiuretic hormone occur to restore the normal blood homeostasis.
These events cause further retention of renal sodium and fluid.
Splanchnic vasodilation increases splanchnic lymph production,
exceeding the lymph transportation system capacity, and leads to
lymph leakage into the peritoneal cavity. Persistent renal sodium
and fluid retention, alongside increased splanchnic vascular
permeability in addition to lymph leakage into the peritoneal
cavity, play a major role in a sustained ascites formation.
[0015] Thiazolidinediones (TZD's), such as rosiglitazone, are
peroxisome proliferator-activated receptor (PPAR) gamma agonist
agents used for the treatment of type-2 diabetes and are widely
prescribed. Unfortunately, fluid retention has emerged as the most
common and serious side-effect of TZD's and has become the most
frequent cause of discontinuation of therapy. The incidence of
TZD-induced fluid retention ranges from 7% in monotherapy and to as
high as 15% when combined with insulin (Yan, T., Soodvilai, S.,
PPAR Research volume 2008, article ID 943614). The mechanisms for
such side-effects are not fully understood but may be related in Na
and fluid re-absorption in the kidney. However TZD-induced fluid
retention is resistant to loop diuretics or thiazide diuretics, and
combination of peroxisome proliferator-activated receptor (PPAR)
alpha with PPAR gamma agonists, which were proposed to reduce such
fluid overload, are associated with major adverse cardiovascular
events.
[0016] In view of the foregoing, it is recognized that salt and
fluid accumulation contribute to the morbidity and mortality of
many diseases, including heart failure (in particular, congestive
heart failure), chronic kidney disease, end-stage renal disease,
liver disease and the like. It is also accepted that salt and fluid
accumulation are risk factors for hypertension. Accordingly, there
is a clear need for a medicament that, when administered to a
patient in need, would result in a reduction in sodium retention,
fluid retention, or preferably both. Such a medicament would more
preferably also not involve or otherwise impair renal mechanisms of
fluid/Na homeostasis.
[0017] One option to consider for treating excessive fluid overload
is to induce diarrhea. Diarrhea may be triggered by several agents
including, for example, laxatives such as sorbitol,
polyethyleneglycol, bisacodyl and phenolphthaleine. Sorbitol and
polyethyleneglycol triggers osmotic diarrhea with low levels of
secreted electrolytes; thus, their utility in removing sodium salt
from the GI tract is limited. The mechanism of action of
phenolphthalein is not clearly established, but is thought to be
caused by inhibition of the Na/K ATPase and the Cl/HCO.sub.3 anion
exchanger and stimulation of electrogenic anion secretion (see,
e.g., Eherer, A. J., C. A. Santa Ana, J. Porter, and J. S.
Fordtran, 1993, Gastroenterology, v. 104, no. 4, p. 1007-1012).
However, some laxatives, such as phenolphthalein, are not viable
options for the chronic treatment of fluid overload, due to the
potential risk of carcinogenicity in humans. Furthermore, laxatives
may not be used chronically, as they have been shown to be an
irritant and cause mucosal damage. Accordingly, it should also be
recognized that the induction of chronic diarrhea as part of an
effort to control salt and fluid overload would be an undesired
treatment modality for most patients. Any medicament utilizing the
GI tract for this purpose would therefore need to control diarrhea
in order to be of practical benefit.
[0018] One approach for the treatment of mild diarrhea is the
administration of a fluid-absorbing polymer, such as the natural
plant fiber psyllium. Polymeric materials, and more specifically
hydrogel polymers, may also be used for the removal of fluid from
the gastrointestinal (GI) tract. The use of such polymers is
described in, for example, U.S. Pat. No. 4,470,975 and No.
6,908,609, the entire contents of which are incorporated herein by
reference for all relevant and consistent purposes. However, for
such polymers to effectively remove significant quantities of
fluid, they must desirably resist the static and osmotic pressure
range existing in the GI tract. Many mammals, including humans,
make a soft feces with a water content of about 70%, and do so by
transporting fluid against the high hydraulic resistance imposed by
the fecal mass. Several studies show that the pressure required to
dehydrate feces from about 80% to about 60% is between about 500
kPa and about 1000 kPa (i.e., about 5 to about 10 atm). (See, e.g.,
McKie, A. T., W. Powrie, and R. J. Naftalin, 1990, Am J Physiol, v.
258, no. 3 Pt 1, p. G391-G394; Bleakman, D., and R. J. Naftalin,
1990, Am J Physiol, v. 258, no. 3 Pt 1, p. G377-G390; Zammit, P.
S., M. Mendizabal, and R. J. Naftalin, 1994, J Physiol, v. 477 (Pt
3), p. 539-548.) However, the static pressure measured
intraluminally is usually between about 6 kPa and about 15 kPa. The
rather high pressure needed to dehydrate feces is essentially due
to an osmotic process and not a mechanical process produced by
muscular forces. The osmotic pressure arises from the active
transport of salt across the colonic mucosa that ultimately
produces a hypertonic fluid absorption. The osmotic gradient
produced drives fluid from the lumen to the serosal side of the
mucosa. Fluid-absorbing polymers, such as those described in for
example U.S. Pat. Nos. 4,470,975 and 6,908,609, may not be able to
sustain such pressure. Such polymers may collapse in a normal colon
where the salt absorption process is intact, hence removing a
modest quantity of fluid and thereby salt.
[0019] Synthetic polymers that bind sodium have also been
described. For example, ion-exchange polymeric resins, such as
Dowex-type cation exchange resins, have been known since about the
1950's. However, with the exception of Kayexalate.TM. (or
Kionex.TM.), which is a polystyrene sulfonate salt approved for the
treatment of hyperkalemia, cation exchange resins have very limited
use as drugs, due at least in part to their limited capacity and
poor cation binding selectivity. Additionally, during the
ion-exchange process, the resins may release a stochiometric amount
of exogenous cations (e.g., H, K, Ca), which may in turn
potentially cause acidosis (H), hyperkalemia (K) or contribute to
vascular calcification (Ca). Such resins may also cause
constipation.
[0020] Gastrointestinal Tract Disorders
[0021] Constipation is characterized by infrequent and difficult
passage of stool and becomes chronic when a patient suffers
specified symptoms for over 12 non-consecutive weeks within a
12-month period. Chronic constipation is idiopathic if it is not
caused by other diseases or by use of medications. An
evidence-based approach to the management of chronic constipation
in North America (Brandt et al., 2005, Am. J. Gastroenterol.
100(Suppl.1):S5-S21) revealed that prevalence is approximately 15%
of the general population. Constipation is reported more commonly
in women, the elderly, non-whites, and individuals from lower
socioeconomic groups.
[0022] Irritable bowel syndrome (IBS) is a common GI disorder
associated with alterations in motility, secretion and visceral
sensation. A range of clinical symptoms characterizes this
disorder, including stool frequency and form, abdominal pain and
bloating. The recognition of clinical symptoms of IBS are yet to be
defined, but it is now common to refer to diarrhea-predominant IBS
(D-IBS) and constipation-predominant IBS (C-IBS), wherein D-IBS is
defined as continuous passage of loose or watery stools and C-IBS
as a group of functional disorders which present as difficult,
infrequent or seemingly incomplete defecation. The pathophysiology
of IBS is not fully understood, and a number of mechanisms have
been suggested. Visceral hypersensitivity is often considered to
play a major etiologic role and has been proposed to be a
biological marker even useful to discriminate IBS from other causes
of abdominal pain. In a recent clinical study (Posserud, I. et al,
Gastroenterology, 2007; 133:1113-1123) IBS patients were submitted
to a visceral sensitivity test (Balloon distention) and compared
with healthy subjects. It revealed that 61% of the IBS patients had
an altered visceral perception as measured by pain and discomfort
threshold. Other reviews have documented the role of visceral
hypersensitivity in abdominal pain symptomatic of various
gastrointestinal tract disorders (Akbar, A, et al, Aliment.
Pharmaco. Ther., 2009, 30, 423-435; Bueno et al.,
Neurogastroenterol Motility (2007) 19 (suppl.1), 89-119). Colonic
and rectal distention have been widely used as a tool to assess
visceral sensitivity in animal and human studies. The type of
stress used to induce visceral sensitivity varies upon the models
(see for instance Eutamen, H Neurogastroenterol Motil. 2009 Aug.
25. [Epub ahead of print]), however stress such as Partial
restraint stress (PRS) is a relatively mild, non-ulcerogenic model
that is considered more representative of the IBS setting.
[0023] Constipation is commonly found in the geriatric population,
particularly patients with osteoporosis who have to take calcium
supplements. Calcium supplements have shown to be beneficial in
ostoporotic patients to restore bone density but compliance is poor
because of calcium-induced constipation effects.
[0024] Opioid-induced constipation (OIC) (also referred to as
opioid-induced bowel dysfunction or opioid bowel dysfuntion (OBD))
is a common adverse effect associated with opioid therapy. OIC is
commonly described as constipation; however, it is a constellation
of adverse gastrointestinal (GI) effects, which also includes
abdominal cramping, bloating, and gastroesophageal reflux. Patients
with cancer may have disease-related constipation, which is usually
worsened by opioid therapy. However, OIC is not limited to cancer
patients. A recent survey of patients taking opioid therapy for
pain of non-cancer origin found that approximately 40% of patients
experienced constipation related to opioid therapy (<3 complete
bowel movements per week) compared with 7.6% in a control group. Of
subjects who required laxative therapy, only 46% of opioid-treated
patients (control subjects, 84%) reported achieving the desired
treatment results >50% of the time (Pappagallo, 2001, Am. J.
Surg. 182(5A Suppl.):11S-18S).
[0025] Some patients suffering from chronic idiopathic constipation
can be successfully treated with lifestyle modification, dietary
changes and increased fluid and fiber intake, and these treatments
are generally tried first. For patients who fail to respond to
these approaches, physicians typically recommend laxatives, most of
which are available over-the-counter. Use of laxatives provided
over-the-counter is judged inefficient by about half of the
patients (Johanson and Kralstein, 2007, Aliment. Pharmacol. Ther.
25(5):599-608). Other therapeutic options currently prescribed or
in clinical development for the treatment of IBS and chronic
constipation including OIC are described in, for example: Chang et
al., 2006, Curr. Teat. Options Gastroenterol. 9(4):314-323; Gershon
and Tack, 2007, Gastroenterology 132(1):397-414; and, Hammerle and
Surawicz, 2008, World J. Gastroenterol. 14(17):2639-2649. Such
treatments include but are not limited to serotonin receptor
ligands, chloride channel activators, opioid receptor antagonists,
guanylate-cyclase receptor agonists and nucleotide P2Y(2) receptor
agonists. Many of these treatment options are inadequate, as they
may be habit forming, ineffective in some patients, may cause long
term adverse effects, or otherwise are less than optimal.
[0026] Na.sup.+/H.sup.+ Exchanger (NHE) Inhibitors
[0027] A major function of the GI tract is to maintain water/Na
homeostasis by absorbing virtually all water and Na to which the GI
tract is exposed. The epithelial layer covering the apical surface
of the mammalian colon is a typical electrolyte-transporting
epithelium, which is able to move large quantities of salt and
water in both directions across the mucosa. For example, each day
the GI tract processes about 9 liters of fluid and about 800 meq of
Na. (See, e.g., Zachos et al., Molecular physiology of intestinal
Na+/H+ exchange; Annu. Rev. Physiol., v. 67, p. 411-443 (2005).)
Only about 1.5 liters of this fluid and about 150 meq of this
sodium originates from ingestion; rather, the majority of the fluid
(e.g., about 7.5 liters) and sodium (about 650 meq) is secreted via
the GI organs as part of digestion. The GI tract therefore
represents a viable target for modulating systemic sodium and fluid
levels.
[0028] Many reviews have been published on the physiology and
secretory and/or absorption mechanisms of the GI tract (see, e.g.,
Kunzelmann et al., Electrolyte transport in the mammalian colon:
mechanisms and implications for disease; Physiol. Rev., v. 82, no.
1, p. 245-289 (2002); Geibel, J. P.; Secretion and absorption by
colonic crypts; Annu. Rev. Physiol, v. 67, p. 471-490 (2005);
Zachos et al., supra; Kiela, P. R. et al., Apical Na+/H+ exchangers
in the mammalian gastrointestinal tract; J. Physiol. Pharmacol., v.
57 Suppl. 7, p. 51-79 (2006)). The two main mechanisms of Na
absorption are electroneutral and electrogenic transport.
Electroneutral transport is essentially due to the Na.sup.+/H.sup.+
antiport NHE (e.g., NHE-3) and is responsible for the bulk of Na
absorption. Electrogenic transport is provided by the epithelium
sodium channel ("ENaC"). Electroneutral transport is located
primarily in the ileal segment and proximal colon and electrogenic
transport is located in the distal colon.
[0029] Plasma membrane NHEs contribute to maintenance of
intracellular pH and volume, transcellular absorption of NaCl and
NaHCO.sub.3, and fluid balance carried out by epithelial cells,
especially in the kidney, intestine, gallbladder, and salivary
glands, as well as regulation of systemic pH. There exists a body
of literature devoted to the role and clinical intervention on
systemic NHEs to treat disorders related to ischemia and
reperfusion for cardioprotection or renal protection. Nine isoforms
of NHEs have been identified (Kiela, P. R., et al.; Apical NA+/H+
exchangers in the mammalian gastrointestinal tract; J. Physiol.
Pharmacol., v. 57 Suppl 7, p. 51-79 (2006)), of which NHE-2, NHE-3
and NHE-8 are expressed on the apical side of the GI tract, with
NHE-3 providing a larger contribution to transport. Another, yet to
be identified, Cl-dependant NHE has been identified in the crypt of
rat cells. In addition, much research has been devoted to
identifying inhibitors of NHEs. The primary targets of such
research have been NHE-1 and NHE-3. Small molecule NHE inhibitors
are, for example, described in: U.S. Pat. Nos. 5,866,610;
6,399,824; 6,911,453; 6,703,405; 6,005,010; 6,736,705; 6,887,870;
6,737,423; 7,326,705; 5,824,691 (WO 94/026709); U.S. Pat. No.
6,399,824 (WO 02/024637); U.S. Pat. Pub. Nos. 2004/0039001 (WO
02/020496); 2005/0020612 (WO 03/055490); 2004/0113396 (WO
03/051866); 2005/0020612; 2005/0054705; 2008/0194621; 2007/0225323;
2004/0039001; 2004/0224965; 2005/0113396; 2007/0135383;
2007/0135385; 2005/0244367; 2007/0270414; International Publication
Nos. WO 01/072742; WO 01/021582 (CA2387529); WO 97/024113
(CA02241531) and European Pat. No. EP0744397 (CA2177007); all of
which are incorporated herein by reference in their entirety for
all relevant and consistent purposes.
[0030] However, such research failed to develop or recognize the
value or importance of NHE inhibitors that are not absorbed (i.e.,
not systemic) and target the gastrointestinal tract, as disclosed
recently in WO 2010/078449. Such inhibitors can be utilized in the
treatment of disorders associated with fluid retention and salt
overload and in the treatment of GI tract disorders, including the
treatment or reduction of pain associated with a gastrointestinal
tract disorder. Such inhibitors are particular advantageous because
they can be delivered with reduced fear of systemic on-target or
off-target effects (e.g., little or no risk of renal involvement or
other systemic effects.
[0031] Accordingly, while progress has been made in the foregoing
fields, there remains a need in the art for novel compounds for use
in the disorders associated with fluid retention and salt overload
and in the treatment of gastrointestinal tract disorders, including
the treatment or reduction of pain associated with a
gastrointestinal tract disorder. The present invention fulfills
this need and provides further related advantages.
BRIEF SUMMARY
[0032] In brief, the present invention is directed to compounds
that are substantially active in the gastrointestinal tract to
inhibit NHE-mediated antiport of sodium ions and hydrogen ions, and
the use of such compounds in the treatment of disorders associated
with fluid retention and salt overload and in the treatment of
gastrointestinal tract disorders, including the treatment or
reduction of pain associated with a gastrointestinal tract
disorder.
[0033] In one embodiment, a compound is provided having the
structure of Formula (I):
##STR00001##
or a stereoisomer, prodrug or pharmaceutically acceptable salt
thereof,
[0034] wherein: [0035] (a) NHE is a NHE-inhibiting small molecule
moiety having the following structure of Formula (A):
##STR00002##
[0036] wherein: [0037] each R.sub.1, R.sub.2, R.sub.3, R.sub.5 and
R.sub.9 are independently selected from H, halogen,
--NR.sub.7(CO)R.sub.8, --(CO)NR.sub.7R.sub.8,
--SO.sub.2--NR.sub.7R.sub.8, --NR.sub.7SO.sub.2R.sub.8,
--NR.sub.7R.sub.8, --OR.sub.7, --SR.sub.7, --O(CO)NR.sub.7R.sub.8,
--NR.sub.7(CO)OR.sub.8, and --NR.sub.7SO.sub.2NR.sub.8, where
R.sub.7 and R.sub.8 are independently selected from H,
C.sub.1-6alkyl, --C.sub.1-6alkyl-OH or a bond linking the
NHE-inhibiting small molecule to L, provided at least one is a bond
linking the NHE-inhibiting small molecule to L; [0038] R.sub.4 is
selected from H, C.sub.1-C.sub.7 alkyl, or a bond linking the
NHE-inhibiting small molecule to L; [0039] R.sub.6 is absent or
selected from H and C.sub.1-C.sub.7 alkyl; and [0040] Ar1 and Ar2
independently represent an aromatic ring or a heteroaromatic ring;
[0041] (b) Core is a Core moiety having the following structure of
Formula (B):
##STR00003##
[0042] wherein: [0043] X is selected from C(X.sub.1), N and
N(C.sub.1-6alkyl); [0044] X.sub.1 is selected from hydrogen,
optionally substituted alkyl, --NX.sub.aX.sub.b, --NO.sub.2,
--NX.sub.c--C(.dbd.O)--NX.sub.c--X.sub.a,
--C(.dbd.O)NX.sub.c--X.sub.a, --NX.sub.c--C(.dbd.O)--X.sub.a,
--NX.sub.c--SO.sub.2--X.sub.a, --C(.dbd.O)--X.sub.a and --OX.sub.a,
[0045] each X.sub.a and X.sub.b are independently selected from
hydrogen, optionally substituted alkyl, optionally substituted
cycloalkyl, optionally substituted cycloalkylalkyl, optionally
substituted heterocyclyl, optionally substituted heterocyclylalkyl,
optionally substituted aryl, optionally substituted aralkyl,
optionally substituted heteroaryl and optionally substituted
heteroarylalkyl; [0046] Y is C.sub.1-6alkylene; [0047] Z is
selected from --NZ.sub.a--C(.dbd.O)--NZ.sub.a--,
--C(.dbd.O)NZ.sub.a--, --NZ.sub.a--C(.dbd.O)-- and heteroaryl when
X is CX.sub.1; [0048] Z is selected from
--NZ.sub.a--C(.dbd.O)--NZ.sub.a--, --NZ.sub.a--C(.dbd.O)-- and
heteroaryl when X is N or N(C.sub.1-6alkyl); and [0049] each
X.sub.c and Z.sub.a is independently selected from hydrogen and
C.sub.1-6alkyl; and [0050] (c) L is a bond or linker connecting the
Core moiety to the NHE-inhibiting small molecule moieties.
[0051] In more specific embodiments, the NHE-inhibiting small
molecule moiety has the following structure:
##STR00004##
[0052] wherein: [0053] each R.sub.1, R.sub.2 and R.sub.3 are
independently selected from H, halogen, --NR.sub.7(CO)R.sub.8,
--(CO)NR.sub.7R.sub.8, --SO.sub.2--NR.sub.7R.sub.8,
--NR.sub.7SO.sub.2R.sub.8, --NR.sub.7R.sub.8, --OR.sub.7,
--SR.sub.7, --O(CO)NR.sub.7R.sub.8, --NR.sub.7(CO)OR.sub.8, and
--NR.sub.7SO.sub.2NR.sub.8, where R.sub.7 and R.sub.8 are
independently selected from H, C.sub.1-6alkyl, --C.sub.1-6alkyl-OH
or a bond linking the NHE-inhibiting small molecule to L, provided
at least one is a bond linking the NHE-inhibiting small molecule to
L.
[0054] In further more specific embodiments, the NHE-inhibiting
small molecule moiety has one of the following structures:
##STR00005##
[0055] In other more specific embodiments, L is a polyalkylene
glycol linker. For example, in certain embodiments, L is a
polyethylene glycol linker.
[0056] In other more specific embodiments, X is C(X.sub.1). In
further embodiments, each X, is hydrogen.
[0057] In other more specific embodiments, X is N.
[0058] In other more specific embodiments, each Z.sub.a is
hydrogen.
[0059] In another embodiment, a compound is provided having the
structure of Formula (II):
##STR00006##
or a stereoisomer, prodrug or pharmaceutically acceptable salt
thereof,
[0060] wherein: [0061] (a) NHE is a NHE-inhibiting small molecule
moiety having the structure of Formula (A):
##STR00007##
[0062] wherein: [0063] each R.sub.1, R.sub.2, R.sub.3, R.sub.5 and
R.sub.9 are independently selected from H, halogen,
--NR.sub.7(CO)R.sub.8, --(CO)NR.sub.7R.sub.8,
--SO.sub.2--NR.sub.7R.sub.8, --NR.sub.7SO.sub.2R.sub.8,
--NR.sub.7R.sub.8, --OR.sub.7, --SR.sub.7, --O(CO)NR.sub.7R.sub.8,
--NR.sub.7(CO)OR.sub.8, and --NR.sub.7SO.sub.2NR.sub.8, where
R.sub.7 and R.sub.8 are independently selected from H,
C.sub.1-6alkyl, --C.sub.1-6alkyl-OH or a bond linking the
NHE-inhibiting small molecule to L, provided at least one is a bond
linking the NHE-inhibiting small molecule to L; [0064] R.sub.4 is
selected from H, C.sub.1-C.sub.7 alkyl, or a bond linking the
NHE-inhibiting small molecule to L; [0065] R.sub.6 is absent or
selected from H and C.sub.1-C.sub.7 alkyl; and [0066] Ar1 and Ar2
independently represent an aromatic ring or a heteroaromatic ring;
[0067] (b) Core is a Core moiety having the following structure of
Formula (C):
##STR00008##
[0068] wherein:
[0069] W is selected from alkylene, polyalkylene glycol,
--C(.dbd.O)--NH-(alkylene)-NH--C(.dbd.O)--,
--C(.dbd.O)--NH-(polyalkylene glycol)-NH--C(.dbd.O)--,
--C(.dbd.O)-(alkylene)-C(.dbd.O)--, --C(.dbd.O)-(polyalkylene
glycol)-C(.dbd.O)-- and cycloalkyl,
[0070] X is N;
[0071] Y is C.sub.1-6alkylene;
[0072] Z is selected from --NZ.sub.a--C(.dbd.O)--NZ.sub.a--,
--C(.dbd.O)NZ.sub.a--, --NZ.sub.a--C(.dbd.O)-- and heteroaryl;
[0073] each Z.sub.a is independently selected from hydrogen and
C.sub.1-6alkyl; and [0074] (c) L is a bond or linker connecting the
Core moiety to the NHE-inhibiting small molecules.
[0075] In more specific embodiments, the NHE-inhibiting small
molecule moiety has the following structure:
##STR00009##
[0076] wherein: [0077] each R.sub.1, R.sub.2 and R.sub.3 are
independently selected from H, halogen, --NR.sub.7(CO)R.sub.8,
--(CO)NR.sub.7R.sub.8, --SO.sub.2--NR.sub.7R.sub.8,
--NR.sub.7SO.sub.2R.sub.8, --NR.sub.7R.sub.8, --OR.sub.7,
--SR.sub.7, --O(CO)NR.sub.7R.sub.8, --NR.sub.7(CO)OR.sub.8, and
--NR.sub.7SO.sub.2NR.sub.8, where R.sub.7 and R.sub.8 are
independently selected from H, C.sub.1-6alkyl, --C.sub.1-6alkyl-OH
or a bond linking the NHE-inhibiting small molecule to L, provided
at least one is a bond linking the NHE-inhibiting small molecule to
L.
[0078] In further more specific embodiments, the NHE-inhibiting
small molecule moiety has one of the following structures:
##STR00010##
[0079] In other more specific embodiments, L is a polyalkylene
glycol linker. For example, in certain embodiments, L is a
polyethylene glycol linker.
[0080] In other more specific embodiments, X is C(X.sub.1). In
further embodiments, each X, is hydrogen.
[0081] In other more specific embodiments, X is N.
[0082] In other more specific embodiments, each Z.sub.a is
hydrogen.
[0083] In another embodiment, a pharmaceutical composition is
provided comprising a compound as set forth above, or a
stereoisomer, pharmaceutically acceptable salt or prodrug thereof,
and a pharmaceutically acceptable carrier, diluent or
excipient.
[0084] In further embodiments, the composition further comprises a
fluid-absorbing polymer. In further embodiments, the
fluid-absorbing polymer is delivered directly to the colon. In
further embodiments, the fluid-absorbing polymer has a fluid
absorbency of at least about 15 g of isotonic fluid per g of
polymer under a static pressure of about 5 kPa. In further
embodiments, the fluid-absorbing polymer has a fluid absorbency of
at least about 15 g of isotonic fluid per g of polymer under a
static pressure of about 10 kPa. In further embodiments, the
fluid-absorbing polymer is characterized by a fluid absorbency of
at least about 10 g/g. In further embodiments, the fluid-absorbing
polymer is characterized by a fluid absorbency of at least about 15
g/g. In further embodiments, the fluid-absorbing polymer is
superabsorbent. In further embodiments, the fluid-absorbing polymer
is a crosslinked, partially neutralized polyelectrolyte hydrogel.
In further embodiments, the fluid-absorbing polymer is a
crosslinked polyacrylate. In further embodiments, the
fluid-absorbing polymer is a polyelectrolyte. In further
embodiments, the fluid-absorbing polymer is calcium Carbophil. In
further embodiments, the fluid-absorbing polymer is prepared by a
high internal phase emulsion process. In further embodiments, the
fluid-absorbing polymer is a foam. In further embodiments, the
fluid-absorbing polymer is prepared by a aqueous free radical
polymerization of acrylamide or a derivative thereof, a crosslinker
and a free radical initiator redox system in water. In further
embodiments, the fluid-absorbing polymer is a hydrogel. In further
embodiments, the fluid-absorbing polymer is an N-alkyl acrylamide.
In further embodiments, the fluid-absorbing polymer is a
superporous gel. In further embodiments, the fluid-absorbing
polymer is naturally occurring. In further embodiments, the
fluid-absorbing polymer is selected from the group consisting of
xanthan, guar, wellan, hemicelluloses, alkyl-cellulose
hydro-alkyl-cellulose, carboxy-alkyl-cellulose, carrageenan,
dextran, hyaluronic acid and agarose. In further embodiments, the
fluid-absorbing polymer is psyllium. In further embodiments, the
fluid-absorbing polymer is a polysaccharide that includes xylose
and arabinose. In further embodiments, the fluid-absorbing polymer
is a polysaccharide that includes xylose and arabinose, wherein the
ratio of xylose to arabinose is at least about 3:1, by weight.
[0085] In further embodiments, the composition further comprises
another pharmaceutically active agent or compound. In further
embodiments, the composition further comprises another
pharmaceutically active agent or compound selected from the group
consisting of a diuretic, cardiac glycoside, ACE inhibitor,
angiotensin-2 receptor antagonist, aldosterone antagonist,
aldosterone synthase inhibitor, renin inhibitor, calcium channel
blocker, beta blocker, alpha blocker, central alpha agonist,
vasodilator, blood thinner, anti-platelet agent, lipid-lowering
agent, and peroxisome proliferator-activated receptor (PPAR) gamma
agonist agent. In further embodiments, the diuretic is selected
from the group consisting of a high ceiling loop diuretic, a
benzothiadiazide diuretic, a potassium sparing diuretic, and a
osmotic diuretic. In further embodiments, the composition further
comprises another pharmaceutically active agent or compound
selected from the group consisting of an analgesic peptide or
agent. In further embodiments, the composition further comprises
another pharmaceutically active agent or compound selected from the
group consisting of a laxative agent selected from a bulk-producing
agent (e.g. psyllium husk (Metamucil)), methylcellulose (Citrucel),
polycarbophil, dietary fiber, apples, stool softeners/surfactant
(e.g., docusate, Colace, Diocto), a hydrating or osmotic agent
(e.g., dibasic sodium phosphate, magnesium citrate, magnesium
hydroxide (Milk of magnesia), magnesium sulfate (which is Epsom
salt), monobasic sodium phosphate, sodium biphosphate), a
hyperosmotic agent (e.g., glycerin suppositories, sorbitol,
lactulose, and polyethylene glycol (PEG)).
[0086] In another embodiment, a method for inhibiting NHE-mediated
antiport of sodium and hydrogen ions is provided, the method
comprising administering to a mammal in need thereof a
pharmaceutically effective amount of a compound or pharmaceutical
composition as set forth above.
[0087] In another embodiment, a method for treating a disorder
associated with fluid retention or salt overload is provided, the
method comprising administering to a mammal in need thereof a
pharmaceutically effective amount of a compound or pharmaceutical
composition as set forth above.
[0088] In another embodiment, a method for treating a disorder
selected from the group consisting of heart failure (such as
congestive heart failure), chronic kidney disease, end-stage renal
disease, liver disease, and peroxisome proliferator-activated
receptor (PPAR) gamma agonist-induced fluid retention is provided,
the method comprising administering to a mammal in need thereof a
pharmaceutically effective amount of a compound or pharmaceutical
composition as set forth above.
[0089] In another embodiment, a method for treating hypertension is
provided, the method comprising administering to a mammal in need
thereof a pharmaceutically effective amount of a compound or
pharmaceutical composition as set forth above.
[0090] In further embodiments, the method comprises administering a
pharmaceutically effective amount of the compound to the mammal in
order to increase the mammal's daily fecal output of sodium and/or
fluid. In further embodiments, the method comprises administering a
pharmaceutically effective amount of the compound to the mammal in
order to increase the mammal's daily fecal output of sodium by at
least about 30 mmol, and/or fluid by at least about 200 ml. In
further embodiments, the mammal's fecal output of sodium and/or
fluid is increased without introducing another type of cation in a
stoichiometric or near stoichiometric fashion via an ion exchange
process. In further embodiments, the method further comprises
administering to the mammal a fluid-absorbing polymer to absorb
fecal fluid resulting from the use of the compound that is
substantially active in the gastrointestinal tract to inhibit
NHE-mediated antiport of sodium ions and hydrogen ions therein.
[0091] In further embodiments, the compound or composition is
administered to treat hypertension. In further embodiments, the
compound or composition is administered to treat hypertension
associated with dietary salt intake. In further embodiments,
administration of the compound or composition allows the mammal to
intake a more palatable diet. In further embodiments, the compound
or composition is administered to treat fluid overload. In further
embodiments, the fluid overload is associated with congestive heart
failure. In further embodiments, the fluid overload is associated
with end stage renal disease. In further embodiments, the fluid
overload is associated with peroxisome proliferator-activated
receptor (PPAR) gamma agonist therapy. In further embodiments, the
compound or composition is administered to treat sodium overload.
In further embodiments, the compound or composition is administered
to reduce interdialytic weight gain in ESRD patients. In further
embodiments, the compound or composition is administered to treat
edema. In further embodiments, the edema is caused by chemotherapy,
pre-menstrual fluid overload or preeclampsia.
[0092] In further embodiments, the compound or composition is
administered orally, by rectal suppository, or enema.
[0093] In further embodiments, the method comprises administering a
pharmaceutically effective amount of the compound or composition in
combination with one or more additional pharmaceutically active
compounds or agents. In further embodiments, the one or more
additional pharmaceutically active compounds or agents is selected
from the group consisting of a diuretic, cardiac glycoside, ACE
inhibitor, angiotensin-2 receptor antagonist, aldosterone
antagonist, aldosterone synthase inhibitor, renin inhibitor,
calcium channel blocker, beta blocker, alpha blocker, central alpha
agonist, vasodilator, blood thinner, anti-platelet agent,
lipid-lowering agent, and peroxisome proliferator-activated
receptor (PPAR) gamma agonist agent. In further embodiments, the
diuretic is selected from the group consisting of a high ceiling
loop diuretic, a benzothiadiazide diuretic, a potassium sparing
diuretic, and a osmotic diuretic. In further embodiments, the
pharmaceutically effective amount of the compound or composition,
and the one or more additional pharmaceutically active compounds or
agents, are administered as part of a single pharmaceutical
preparation. In further embodiments, the pharmaceutically effective
amount of the compound or composition, and the one or more
additional pharmaceutically active compounds or agents, are
administered as individual pharmaceutical preparations. In further
embodiments, the individual pharmaceutical preparation are
administered sequentially. In further embodiments, the individual
pharmaceutical preparation are administered simultaneously.
[0094] In another embodiment, a method for treating a
gastrointestinal tract disorder is provided, the method comprising
administering to a mammal in need thereof a pharmaceutically
effective amount of a compound or pharmaceutical composition as set
forth above.
[0095] In further embodiments, the gastrointestinal tract disorder
is a gastrointestinal motility disorder. In further embodiments,
the gastrointestinal tract disorder is irritable bowel syndrome. In
further embodiments, the gastrointestinal tract disorder is chronic
constipation. In further embodiments, the gastrointestinal tract
disorder is chronic idiopathic constipation. In further
embodiments, the gastrointestinal tract disorder is chronic
constipation occurring in cystic fibrosis patients. In further
embodiments, the gastrointestinal tract disorder is opioid-induced
constipation. In further embodiments, the gastrointestinal tract
disorder is a functional gastrointestinal tract disorder. In
further embodiments, the gastrointestinal tract disorder is
selected from the group consisting of chronic intestinal
pseudo-obstruction and colonic pseudo-obstruction. In further
embodiments, the gastrointestinal tract disorder is Crohn's
disease. In further embodiments, the gastrointestinal tract
disorder is ulcerative colitis. In further embodiments, the
gastrointestinal tract disorder is a disease referred to as
inflammatory bowel disease. In further embodiments, the
gastrointestinal tract disorder is associated with chronic kidney
disease (stage 4 or 5). In further embodiments, the
gastrointestinal tract disorder is constipation induced by calcium
supplement. In further embodiments, the gastrointestinal tract
disorder is constipation, and the constipation to be treated is
associated with the use of a therapeutic agent. In further
embodiments, the gastrointestinal tract disorder is constipation,
and the constipation to be treated is associated with a neuropathic
disorder. In further embodiments, the gastrointestinal tract
disorder is constipation, and the constipation to be treated is
post-surgical constipation (postoperative ileus). In further
embodiments, the gastrointestinal tract disorder is constipation,
and the constipation to be treated is idiopathic (functional
constipation or slow transit constipation). In further embodiments,
the gastrointestinal tract disorder is constipation, and the
constipation to be treated is associated with neuropathic,
metabolic or an endocrine disorder (e.g., diabetes mellitus, renal
failure, hypothyroidism, hyperthyroidism, hypocalcaemia, Multiple
Sclerosis, Parkinson's disease, spinal cord lesions,
neurofibromatosis, autonomic neuropathy, Chagas disease,
Hirschsprung's disease or cystic fibrosis, and the like). In
further embodiments, the gastrointestinal tract disorder is
constipation, and the constipation to be treated is due the use of
drugs selected from analgesics (e.g., opioids), antihypertensives,
anticonvulsants, antidepressants, antispasmodics and
antipsychotics.
[0096] In another embodiment, a method for treating irritable bowel
syndrome is provided, the method comprising administering to a
mammal in need thereof a pharmaceutically effective amount of a
compound or pharmaceutical composition as set forth above.
[0097] In further embodiments of the above embodiments, the
compound or composition is administered to treat or reduce pain
associated with a gastrointestinal tract disorder. In further
embodiments, the compound or composition is administered to treat
or reduce visceral hypersensitivity associated with a
gastrointestinal tract disorder. In further embodiments, the
compound or composition is administered to treat or reduce
inflammation of the gastrointestinal tract. In further embodiments,
the compound or composition is administered to reduce
gastrointestinal transit time.
[0098] In further embodiments, the compound or composition is
administered either orally or by rectal suppository.
[0099] In further embodiments, the method comprises administering a
pharmaceutically effective amount of the compound or composition,
in combination with one or more additional pharmaceutically active
compounds or agents. In further embodiments, the one or more
additional pharmaceutically active agents or compounds are an
analgesic peptide or agent. In further embodiments, the one or more
additional pharmaceutically active agents or compounds are selected
from the group consisting of a laxative agent selected from a
bulk-producing agent (e.g. psyllium husk (Metamucil)),
methylcellulose (Citrucel), polycarbophil, dietary fiber, apples,
stool softeners/surfactant (e.g., docusate, Colace, Diocto), a
hydrating or osmotic agent (e.g., dibasic sodium phosphate,
magnesium citrate, magnesium hydroxide (Milk of magnesia),
magnesium sulfate (which is Epsom salt), monobasic sodium
phosphate, sodium biphosphate), and a hyperosmotic agent (e.g.,
glycerin suppositories, sorbitol, lactulose, and polyethylene
glycol (PEG)). In further embodiments, the pharmaceutically
effective amount of the compound or composition, and the one or
more additional pharmaceutically active compounds or agents, are
administered as part of a single pharmaceutical preparation. In
further embodiments, the pharmaceutically effective amount of the
compound or composition, and the one or more additional
pharmaceutically active compounds or agents, are administered as
individual pharmaceutical preparations. In further embodiments, the
individual pharmaceutical preparation are administered
sequentially. In further embodiments, the individual pharmaceutical
preparation are administered simultaneously.
[0100] These and other aspects of the invention will be apparent
upon reference to the following detailed description.
DETAILED DESCRIPTION
[0101] In accordance with the present disclosure, and as further
detailed herein below, it has been found that the inhibition of
NHE-mediated antiport of sodium ions (Na.sup.+) and hydrogen ions
(H.sup.+) in the gastrointestinal tract, and more particularly the
gastrointestinal epithelia, is a powerful approach to the treatment
of various disorders that may be associated with or caused by fluid
retention and/or salt overload, and/or disorders such as heart
failure (in particular, congestive heart failure), chronic kidney
disease, end-stage renal disease, liver disease, and/or peroxisome
proliferator-activated receptor (PPAR) gamma agonist-induced fluid
retention. More specifically, it has been found that the inhibition
of the NHE-mediated antiport of sodium ions and hydrogen ions in
the GI tract increases the fecal excretion of sodium, effectively
reducing systemic levels of sodium and fluid. This, in turn,
improves the clinical status of a patient suffering from, for
example, CHF, ESRD/CKD and/or liver disease. It has further been
found that such a treatment may optionally be enhanced by the
co-administration of other beneficial compounds or compositions,
such as for example a fluid-absorbing polymer. The fluid-absorbing
polymer may optimally be chosen so that it does not block or
otherwise negatively interfere with the mechanism of action of the
co-dosed NHE-inhibiting compound.
[0102] Additionally, and also as further detailed herein below, it
has further been found that the inhibition of NHE-mediated antiport
of sodium ions (Na.sup.+) and hydrogen ions (H.sup.+) in the
gastrointestinal tract, and more particularly the gastrointestinal
epithelia, is a powerful approach to the treatment of hypertension,
that may be associated with or caused by fluid retention and/or
salt overload. More specifically, it has been found that the
inhibition of the NHE-mediated antiport of sodium ions and hydrogen
ions in the GI tract increases the fecal excretion of sodium,
effectively reducing systemic levels of sodium and fluid. This, in
turn, improves the clinical status of a patient suffering from
hypertension. Such a treatment may optionally be enhanced by the
co-administration of other beneficial compounds or compositions,
such as for example a fluid-absorbing polymer. The fluid-absorbing
polymer may optimally be chosen so that it does not block or
otherwise negatively interfere with the mechanism of action of the
co-dosed NHE-inhibiting compound.
[0103] Additionally, and also as further detailed herein below, it
has further been found that the inhibition of NHE-mediated antiport
of sodium ions (Na.sup.+) and hydrogen ions (H.sup.+) in the
gastrointestinal tract, and more particularly the gastrointestinal
epithelia, is a powerful approach to the treatment of various
gastrointestinal tract disorders, including the treatment or
reduction of pain associated with gastrointestinal tract disorders,
and more particularly to the restoration of appropriate fluid
secretion in the gut and the improvement of pathological conditions
encountered in constipation states. Applicants have further
recognized that by blocking sodium ion re-absorption, the compounds
of the present disclosure restore fluid homeostasis in the GI
tract, particularly in situations wherein fluid
secretion/absorption is altered in such a way that it results in a
high degree of feces dehydration, low gut motility, and/or a slow
transit-time producing constipation states and GI discomfort
generally. It has further been found that such a treatment may
optionally be enhanced by the co-administration of other beneficial
compounds or compositions, such as for example a fluid-absorbing
polymer. The fluid-absorbing polymer may optimally be chosen so
that it does not block or otherwise negatively interfere with the
mechanism of action of the co-dosed NHE-inhibiting compound.
[0104] Due to the presence of NHEs in other organs or tissues in
the body, the method of the present disclosure employs the use of
compounds and compositions that are desirably highly selective or
localized, thus acting substantially in the gastrointestinal tract
without exposure to other tissues or organs. In this way, any
systemic effects can be minimized (whether they are on-target or
off-target). Accordingly, it is to be noted that, as used herein,
and as further detailed elsewhere herein, "substantially active in
the gastrointestinal tract" generally refers to compounds that are
substantially systemically non-bioavailable and/or substantially
impermeable to the layer of epithelial cells, and more specifically
epithelium of the GI tract. It is to be further noted that, as used
herein, and as further detailed elsewhere herein, "substantially
impermeable" more particularly encompasses compounds that are
impermeable to the layer of epithelial cells, and more specifically
the gastrointestinal epithelium (or epithelial layer).
"Gastrointestinal epithelium" refers to the membranous tissue
covering the internal surface of the gastrointestinal tract.
Accordingly, by being substantially impermeable, a compound has
very limited ability to be transferred across the gastrointestinal
epithelium, and thus contact other internal organs (e.g., the
brain, heart, liver, etc.). The typical mechanism by which a
compound can be transferred across the gastrointestinal epithelium
is by either transcellular transit (a substance travels through the
cell, mediated by either passive or active transport passing
through both the apical and basolateral membranes) and/or by
paracellular transit, where a substance travels between cells of an
epithelium, usually through highly restrictive structures known as
"tight junctions".
[0105] The compounds of the present disclosure may therefore not be
absorbed, and are thus essentially not systemically bioavailable at
all (e.g., impermeable to the gastrointestinal epithelium at all),
or they show no detectable concentration of the compound in serum.
Alternatively, the compounds may: (i) exhibit some detectable
permeability to the layer of epithelial cells, and more
particularly the epithelium of the GI tract, of less than about 20%
of the administered compound (e.g., less than about 15%, about 10%,
or even about 5%, and for example greater than about 0.5%, or 1%),
but then are rapidly cleared in the liver (i.e., hepatic
extraction) via first-pass metabolism; and/or (ii) exhibit some
detectable permeability to the layer of epithelial cells, and more
particularly the epithelium of the GI tract, of less than about 20%
of the administered compound (e.g., less than about 15%, about 10%,
or even about 5%, and for example greater than about 0.5%, or 1%),
but then are rapidly cleared in the kidney (i.e., renal
excretion).
[0106] Compounds may also be cleared from circulation unchanged
into the bile by biliary excretion. The compounds of the present
disclosure may therefore not exhibit detectable concentrations in
the bile. Alternatively, the compounds may exhibit some detectable
concentration in the bile and more particularly the epithelium of
the biliary tract and gallbladder of 10 .mu.M, less than 1 .mu.M,
less than 0.1 .mu.M, less than 0.01 .mu.M or less than about 0.001
.mu.M.
[0107] In this regard it is to be still further noted that, as used
herein, "substantially systemically non-bioavailable" generally
refers to the inability to detect a compound in the systemic
circulation of an animal or human following an oral dose of the
compound. For a compound to be bioavailable, it must be transferred
across the gastrointestinal epithelium (that is, substantially
permeable as defined above), be transported via the portal
circulation to the liver, avoid substantial metabolism in the
liver, and then be transferred into systemic circulation.
[0108] Without being being held to any particular theory, the
NHE-inhibiting compounds (e.g., NHE-3, -2 and/or -8 inhibitors) of
the present disclosure are believed to act via a distinct and
unique mechanism, causing the retention of fluid and ions in the GI
tract (and stimulating fecal excretion) rather than stimulating
increased secretion of said fluid and ions. For example,
lubiprostone (Amitiza.RTM. Sucampo/Takeda) is a bicyclic fatty acid
prostaglandin E1 analog that activates the Type 2 Chloride Channel
(ClC-2) and increases chloride-rich fluid secretion from the
serosal to the mucosal side of the GI tract (see, e.g.,
Pharmacological Reviews for Amitiza.RTM., NDA package). Linaclotide
(MD-1100 acetate, Microbia/Forest Labs) is a 14 amino acid peptide
analogue of an endogenous hormone, guanylin, and indirectly
activates the Cystic Fibrosis Transmembrane Conductance Regulator
(CFTR) thereby inducing fluid and electrolyte secretion into the GI
(see, e.g., Li et al., J. Exp. Med., vol. 202 (2005), pp. 975-986).
The substantially impermeable NHE-inhibiting compounds of the
present disclosure act to inhibit the reuptake of salt and fluid
rather than promote secretion. Since the GI tract processes about 9
liters of fluid and about 800 meq of Na each day, it is anticipated
that NHE inhibition could permit the removal of substantial
quantities of systemic fluid and sodium to resorb edema and resolve
CHF symptoms.
I. Substantially Impermeable or Substantially Systemically
Non-Bioavailable NHE-Inhibiting Compounds
[0109] In one embodiment, a compound is provided having the
structure of Formula (I):
##STR00011##
or a stereoisomer, prodrug or pharmaceutically acceptable salt
thereof,
[0110] wherein: [0111] (a) NHE is a NHE-inhibiting small molecule
moiety having the following structure of Formula (A):
##STR00012##
[0112] wherein: [0113] each R.sub.1, R.sub.2, R.sub.3, R.sub.5 and
R.sub.9 are independently selected from H, halogen,
--NR.sub.7(CO)R.sub.8, --(CO)NR.sub.7R.sub.8,
--SO.sub.2--NR.sub.7R.sub.8, --NR.sub.7SO.sub.2R.sub.8,
--NR.sub.7R.sub.8, --OR.sub.7, --SR.sub.7, --O(CO)NR.sub.7R.sub.8,
--NR.sub.7(CO)OR.sub.8, and --NR.sub.7SO.sub.2NR.sub.8, where
R.sub.7 and R.sub.8 are independently selected from H,
C.sub.1-6alkyl, --C.sub.1-6alkyl-OH or a bond linking the
NHE-inhibiting small molecule to L, provided at least one is a bond
linking the NHE-inhibiting small molecule to L; [0114] R.sub.4 is
selected from H, C.sub.1-C.sub.7 alkyl, or a bond linking the
NHE-inhibiting small molecule to L; [0115] R.sub.6 is absent or
selected from H and C.sub.1-C.sub.7 alkyl; and [0116] Ar1 and Ar2
independently represent an aromatic ring or a heteroaromatic ring;
[0117] (b) Core is a Core moiety having the following structure of
Formula (B):
##STR00013##
[0118] wherein: [0119] X is selected from C(X.sub.1), N and
N(C.sub.1-6alkyl); [0120] X.sub.1 is selected from hydrogen,
optionally substituted alkyl, --NX.sub.aX.sub.b, --NO.sub.2,
--NX.sub.c--C(.dbd.O)--NX.sub.c--X.sub.a,
--C(.dbd.O)NX.sub.c--X.sub.a, --NX.sub.c--C(.dbd.O)--X.sub.a,
--NX.sub.c--SO.sub.2--X.sub.a, --C(.dbd.O)--X.sub.a and --OX.sub.a,
[0121] each X.sub.a and X.sub.b are independently selected from
hydrogen, optionally substituted alkyl, optionally substituted
cycloalkyl, optionally substituted cycloalkylalkyl, optionally
substituted heterocyclyl, optionally substituted heterocyclylalkyl,
optionally substituted aryl, optionally substituted aralkyl,
optionally substituted heteroaryl and optionally substituted
heteroarylalkyl; [0122] Y is C.sub.1-6alkylene; [0123] Z is
selected from --NZ.sub.a--C(.dbd.O)--NZ.sub.a--,
--C(.dbd.O)NZ.sub.a--, --NZ.sub.a--C(.dbd.O)-- and heteroaryl when
X is CX.sub.1; [0124] Z is selected from
--NZ.sub.a--C(.dbd.O)--NZ.sub.a--, --NZ.sub.a--C(.dbd.O)-- and
heteroaryl when X is N or N(C.sub.1-6alkyl); and [0125] each
X.sub.c and Z.sub.a is independently selected from hydrogen and
C.sub.1-6alkyl; and [0126] (c) L is a bond or linker connecting the
Core moiety to the NHE-inhibiting small molecule moieties, the
resulting NHE-inhibiting compound (i.e., a compound of Formula (I))
possessing overall physicochemical properties that render it
substantially impermeable or substantially systemically
non-bioavailable. The Core moiety may be bound to essentially any
position on, or within, the NHE-inhibiting small molecule moiety,
provided that the installation thereof does not significantly
adversely impact NHE-inhibiting activity.
[0127] In another embodiment, a compound is provided having the
structure of Formula (II):
##STR00014##
or a stereoisomer, prodrug or pharmaceutically acceptable salt
thereof,
[0128] wherein: [0129] (a) NHE is a NHE-inhibiting small molecule
moiety having the structure of Formula (A):
##STR00015##
[0130] wherein: [0131] each R.sub.1, R.sub.2, R.sub.3, R.sub.5 and
R.sub.9 are independently selected from H, halogen,
--NR.sub.7(CO)R.sub.8, --(CO)NR.sub.7R.sub.8,
--SO.sub.2--NR.sub.7R.sub.8, --NR.sub.7SO.sub.2R.sub.8,
--NR.sub.7R.sub.8, --OR.sub.7, --SR.sub.7, --O(CO)NR.sub.7R.sub.8,
--NR.sub.7(CO)OR.sub.8, and --NR.sub.7SO.sub.2NR.sub.8, where
R.sub.7 and R.sub.8 are independently selected from H,
C.sub.1-6alkyl, --C.sub.1-6alkyl-OH or a bond linking the
NHE-inhibiting small molecule to L, provided at least one is a bond
linking the NHE-inhibiting small molecule to L; [0132] R.sub.4 is
selected from H, C.sub.1-C.sub.7 alkyl, or a bond linking the
NHE-inhibiting small molecule to L; [0133] R.sub.6 is absent or
selected from H and C.sub.1-C.sub.7 alkyl; and [0134] An and Ar2
independently represent an aromatic ring or a heteroaromatic ring;
[0135] (b) Core is a Core moiety having the following structure of
Formula (C):
##STR00016##
[0136] wherein: [0137] W is selected from alkylene, polyalkylene
glycol, --C(.dbd.O)--NH-(alkylene)-NH--C(.dbd.O)--,
--C(.dbd.O)--NH-(polyalkylene glycol)-NH--C(.dbd.O)--,
--C(.dbd.O)-(alkylene)-C(.dbd.O)--, --C(.dbd.O)-(polyalkylene
glycol)-C(.dbd.O)-- and cycloalkyl, [0138] X is N; [0139] Y is
C.sub.1-6alkylene; [0140] Z is selected from
--NZ.sub.a--C(.dbd.O)--NZ.sub.a--, --C(.dbd.O)NZ.sub.a--,
--NZ.sub.a--C(.dbd.O)-- and heteroaryl; [0141] each Z.sub.a is
independently selected from hydrogen and C.sub.1-6alkyl; and [0142]
(c) L is a bond or linker connecting the Core moiety to the
NHE-inhibiting small molecules, the resulting NHE-inhibiting
compound (i.e., a compound of Formula (II)) possessing overall
physicochemical properties that render it substantially impermeable
or substantially systemically non-bioavailable. The Core moiety may
be bound to essentially any position on, or within, the
NHE-inhibiting small molecule moiety, provided that the
installation thereof does not significantly adversely impact
NHE-inhibiting activity.
[0143] It is to be noted that, in the structures illustrated
herein, all of the various linkages or bonds will not be shown in
every instance. For example, in one or more of the structures
illustrated above, a bond or connection between the NHE-inhibiting
small molecule moiety and the Core moiety is not always shown.
However, this should not be viewed in a limiting sense. Rather, it
is to be understood that the NHE-inhibiting small molecule moiety
is bound or connected in some way (e.g., by a bond or linker of
some kind) to the Core moiety, such that the resulting
NHE-inhibiting compound is suitable for use (i.e., substantially
impermeable or substantially systemically non-bioavailable in the
GI tract).
[0144] The above noted embodiments are further illustrated herein
below. For example, the first representation below of an exemplary
oligomer compound, wherein the various parts of the compound are
identified, is intended to provide a broad context for the
disclosure provided herein. It is to be noted that while each
NHE-inhibiting small molecule moiety in the structure below is the
same, it is within the scope of this disclosure that each is
independently selected and may be the same or different. In the
illustration below, the linker moiety is a polyethylene glycol
(PEG) motif PEG derivatives are advantageous due in part to their
aqueous solubility, which may help avoid hydrophobic collapse (the
intramolecular interaction of hydrophobic motifs that can occur
when a hydrophobic molecule is exposed to an aqueous environment
(see, e.g., Wiley, R. A.; Rich, D. H. Medical Research Reviews
1993, 13(3), 327-384). The core moiety illustrated below is also
advantageous because it provides some rigidity to the molecule,
allowing an increase in distance between the NHE-inhibiting small
molecule moieties while minimally increasing rotational degrees of
freedom.
##STR00017##
[0145] In designing and making the substantially impermeable or
substantially systemically non-bioavailable NHE-inhibiting
compounds that may be utilized for the treatments detailed in the
instant disclosure, it may in some cases be advantageous to first
determine a likely point of attachment on a NHE-inhibiting small
molecule moiety, where a core or linker might be installed or
attached before making a series of candidate multivalent or
polyvalent compounds. This may be done by one skilled in the art
via known methods by systematically installing functional groups,
or functional groups displaying a fragment of the desired core or
linker, onto various positions of the NHE-inhibiting small molecule
moiety and then testing these adducts to determine whether the
modified compound still retains desired biological properties
(e.g., NHE-inhibiting activity). An understanding of the SAR of the
compound also allows the design of cores and/or linkers that
contribute positively to the activity of the resulting
compounds.
[0146] Another aspect to be considered in the design of cores and
linkers is the limiting or preventing of hydrophobic collapse.
Compounds with extended hydrocarbon functionalities may collapse
upon themselves in an intramolecular fashion, causing an increased
enthalpic barrier for interaction with the desired biological
target. Accordingly, when designing cores and linkers, these are
preferably designed to be resistant to hydrophobic collapse. For
example, conformational constraints such as rigid monocyclic,
bicyclic or polycyclic rings can be installed in a core or linker
to increase the rigidity of the structure. Unsaturated bonds, such
as alkenes and alkynes, may also or alternatively be installed.
Such modifications may ensure the NHE-inhibiting compound is
accessible for productive binding with its target. Furthermore, the
hydrophilicity of the linkers may be improved by adding hydrogen
bond donor or acceptor motifs, or ionic motifs such as amines that
are protonated in the GI, or acids that are deprotonated. Such
modifications will increase the hydrophilicity of the core or
linker and help prevent hydrophobic collapse. Furthermore, such
modifications will also contribute to the impermeability of the
resulting compounds by increasing tPSA.
[0147] It is understood that any embodiment of the compounds of the
present invention, as set forth above, and any specific substituent
set forth herein in such compounds, as set forth above, may be
independently combined with other embodiments and/or substituents
of such compounds to form embodiments of the inventions not
specifically set forth above. In addition, in the event that a list
of substituents is listed for any particular substituent in a
particular embodiment and/or claim, it is understood that each
individual substituent may be deleted from the particular
embodiment and/or claim and that the remaining list of substituents
will be considered to be within the scope of the invention.
Furthermore, it is understood that in the present description,
combinations of substituents and/or variables of the depicted
formulae are permissible only if such contributions result in
stable compounds.
II. Terminology, Physical and Performance Properties
A. Terminology
[0148] Unless the context requires otherwise, throughout the
present specification and claims, the word "comprise" and
variations thereof, such as, "comprises" and "comprising" are to be
construed in an open, inclusive sense, that is as "including, but
not limited to".
[0149] Reference throughout this specification to "one embodiment"
or "an embodiment" means that a particular feature, structure or
characteristic described in connection with the embodiment is
included in at least one embodiment of the present invention. Thus,
the appearances of the phrases "in one embodiment" or "in an
embodiment" in various places throughout this specification are not
necessarily all referring to the same embodiment. Furthermore, the
particular features, structures, or characteristics may be combined
in any suitable manner in one or more embodiments.
[0150] "Amino" refers to the --NH.sub.2 radical.
[0151] "Cyano" refers to the --CN radical.
[0152] "Hydroxy" or "hydroxyl" refers to the --OH radical.
[0153] "Imino" refers to the .dbd.NH substituent.
[0154] "Nitro" refers to the --NO.sub.2 radical.
[0155] "Oxo" refers to the .dbd.O substituent.
[0156] "Thioxo" refers to the .dbd.S substituent.
[0157] "Alkyl" refers to a straight or branched hydrocarbon chain
radical consisting solely of carbon and hydrogen atoms, which is
saturated or unsaturated (i.e., contains one or more double and/or
triple bonds), having from one to twelve carbon atoms
(C.sub.1-C.sub.12 alkyl), preferably one to eight carbon atoms
(C.sub.1-C.sub.8 alkyl) or one to six carbon atoms (C.sub.1-C.sub.6
alkyl), and which is attached to the rest of the molecule by a
single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl
(iso-propyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl),
3-methylhexyl, 2-methylhexyl, ethenyl, prop-1-enyl, but-1-enyl,
pent-1-enyl, penta-1,4-dienyl, ethynyl, propynyl, butynyl,
pentynyl, hexynyl, and the like. Unless stated otherwise
specifically in the specification, an alkyl group may be optionally
substituted.
[0158] "Alkylene" or "alkylene chain" refers to a straight or
branched divalent hydrocarbon chain linking the rest of the
molecule to a radical group, consisting solely of carbon and
hydrogen, which is saturated or unsaturated (i.e., contains one or
more double and/or triple bonds), and having from one to twelve
carbon atoms, e.g., methylene, ethylene, propylene, n-butylene,
ethenylene, propenylene, n-butenylene, propynylene, n-butynylene,
and the like. The alkylene chain is attached to the rest of the
molecule through a single or double bond and to the radical group
through a single or double bond. The points of attachment of the
alkylene chain to the rest of the molecule and to the radical group
can be through one carbon or any two carbons within the chain.
Unless stated otherwise specifically in the specification, an
alkylene chain may be optionally substituted.
[0159] "Alkoxy" refers to a radical of the formula --OR.sub.a where
R.sub.a is an alkyl radical as defined above containing one to
twelve carbon atoms. Unless stated otherwise specifically in the
specification, an alkoxy group may be optionally substituted.
[0160] "Alkylamino" refers to a radical of the formula --NHR.sub.a
or --NR.sub.aR.sub.a where each R.sub.a is, independently, an alkyl
radical as defined above containing one to twelve carbon atoms.
Unless stated otherwise specifically in the specification, an
alkylamino group may be optionally substituted.
[0161] "Thioalkyl" refers to a radical of the formula --SR.sub.a
where R.sub.a is an alkyl radical as defined above containing one
to twelve carbon atoms. Unless stated otherwise specifically in the
specification, a thioalkyl group may be optionally substituted.
[0162] "Aryl" refers to a hydrocarbon ring system radical
comprising hydrogen, 6 to 18 carbon atoms and at least one aromatic
ring. For purposes of this invention, the aryl radical may be a
monocyclic, bicyclic, tricyclic or tetracyclic ring system, which
may include fused or bridged ring systems. Aryl radicals include,
but are not limited to, aryl radicals derived from aceanthrylene,
acenaphthylene, acephenanthrylene, anthracene, azulene, benzene,
chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane,
indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene,
and triphenylene. Unless stated otherwise specifically in the
specification, the term "aryl" or the prefix "ar-" (such as in
"aralkyl") is meant to include aryl radicals that are optionally
substituted.
[0163] "Aralkyl" refers to a radical of the formula
--R.sub.b--R.sub.e where R.sub.b is an alkylene chain as defined
above and R.sub.e is one or more aryl radicals as defined above,
for example, benzyl, diphenylmethyl and the like. Unless stated
otherwise specifically in the specification, an aralkyl group may
be optionally substituted.
[0164] "Cycloalkyl" or "carbocyclic ring" refers to a stable
non-aromatic monocyclic or polycyclic hydrocarbon radical
consisting solely of carbon and hydrogen atoms, which may include
fused or bridged ring systems, having from three to fifteen carbon
atoms, preferably having from three to ten carbon atoms, and which
is saturated or unsaturated and attached to the rest of the
molecule by a single bond. Monocyclic radicals include, for
example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, and cyclooctyl. Polycyclic radicals include, for
example, adamantyl, norbornyl, decalinyl,
7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwise
stated specifically in the specification, a cycloalkyl group may be
optionally substituted.
[0165] "Cycloalkylalkyl" refers to a radical of the formula
--R.sub.bR.sub.d where R.sub.d is an alkylene chain as defined
above and R.sub.g is a cycloalkyl radical as defined above. Unless
stated otherwise specifically in the specification, a
cycloalkylalkyl group may be optionally substituted.
[0166] "Fused" refers to any ring structure described herein which
is fused to an existing ring structure in the compounds of the
invention. When the fused ring is a heterocyclyl ring or a
heteroaryl ring, any carbon atom on the existing ring structure
which becomes part of the fused heterocyclyl ring or the fused
heteroaryl ring may be replaced with a nitrogen atom.
[0167] "Halo" or "halogen" refers to bromo, chloro, fluoro or
iodo.
[0168] "Haloalkyl" refers to an alkyl radical, as defined above,
that is substituted by one or more halo radicals, as defined above,
e.g., trifluoromethyl, difluoromethyl, trichloromethyl,
2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl,
1,2-dibromoethyl, and the like. Unless stated otherwise
specifically in the specification, a haloalkyl group may be
optionally substituted.
[0169] "Heterocyclyl" or "heterocyclic ring" refers to a stable 3-
to 18-membered non-aromatic ring radical which consists of two to
twelve carbon atoms and from one to six heteroatoms selected from
the group consisting of nitrogen, oxygen and sulfur. Unless stated
otherwise specifically in the specification, the heterocyclyl
radical may be a monocyclic, bicyclic, tricyclic or tetracyclic
ring system, which may include fused or bridged ring systems; and
the nitrogen, carbon or sulfur atoms in the heterocyclyl radical
may be optionally oxidized; the nitrogen atom may be optionally
quaternized; and the heterocyclyl radical may be partially or fully
saturated. Examples of such heterocyclyl radicals include, but are
not limited to, dioxolanyl, thienyl[1,3]dithianyl,
decahydroisoquinolyl, imidazolinyl, imidazolidinyl,
isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl,
octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl,
2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl,
4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl,
thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl,
thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and
1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in
the specification, Unless stated otherwise specifically in the
specification, a heterocyclyl group may be optionally
substituted.
[0170] "N-heterocyclyl" refers to a heterocyclyl radical as defined
above containing at least one nitrogen and where the point of
attachment of the heterocyclyl radical to the rest of the molecule
is through a nitrogen atom in the heterocyclyl radical. Unless
stated otherwise specifically in the specification, a
N-heterocyclyl group may be optionally substituted.
[0171] "Heterocyclylalkyl" refers to a radical of the formula
--R.sub.bR.sub.e where R.sub.b is an alkylene chain as defined
above and R.sub.e is a heterocyclyl radical as defined above, and
if the heterocyclyl is a nitrogen-containing heterocyclyl, the
heterocyclyl may be attached to the alkyl radical at the nitrogen
atom. Unless stated otherwise specifically in the specification, a
heterocyclylalkyl group may be optionally substituted.
[0172] "Heteroaryl" refers to a 5- to 14-membered ring system
radical comprising hydrogen atoms, one to thirteen carbon atoms,
one to six heteroatoms selected from the group consisting of
nitrogen, oxygen and sulfur, and at least one aromatic ring. For
purposes of this invention, the heteroaryl radical may be a
monocyclic, bicyclic, tricyclic or tetracyclic ring system, which
may include fused or bridged ring systems; and the nitrogen, carbon
or sulfur atoms in the heteroaryl radical may be optionally
oxidized; the nitrogen atom may be optionally quaternized. Examples
include, but are not limited to, azepinyl, acridinyl,
benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl,
benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl,
benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl,
benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl,
benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl
(benzothiophenyl), benzotriazolyl,
benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,
dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl,
isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl,
isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl,
isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl,
oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl,
1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl,
phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl,
pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl,
pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl,
isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl,
triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl).
Unless stated otherwise specifically in the specification, a
heteroaryl group may be optionally substituted.
[0173] "N-heteroaryl" refers to a heteroaryl radical as defined
above containing at least one nitrogen and where the point of
attachment of the heteroaryl radical to the rest of the molecule is
through a nitrogen atom in the heteroaryl radical. Unless stated
otherwise specifically in the specification, an N-heteroaryl group
may be optionally substituted.
[0174] "Heteroarylalkyl" refers to a radical of the formula
--R.sub.bR.sub.f where R.sub.b is an alkylene chain as defined
above and R.sub.f is a heteroaryl radical as defined above. Unless
stated otherwise specifically in the specification, a
heteroarylalkyl group may be optionally substituted.
[0175] The term "substituted" used herein means any of the above
groups (i.e., alkyl, alkylene, alkoxy, alkylamino, thioalkyl, aryl,
aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl,
N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl and/or
heteroarylalkyl) wherein at least one hydrogen atom is replaced by
a bond to a non-hydrogen atoms such as, but not limited to: a
halogen atom such as F, Cl, Br, and I; an oxygen atom in groups
such as hydroxyl groups, alkoxy groups, and ester groups; a sulfur
atom in groups such as thiol groups, thioalkyl groups, sulfone
groups, sulfonyl groups, and sulfoxide groups; a nitrogen atom in
groups such as amines, amides, alkylamines, dialkylamines,
arylamines, alkylarylamines, diarylamines, N-oxides, imides, and
enamines; a silicon atom in groups such as trialkylsilyl groups,
dialkylarylsilyl groups, alkyldiarylsilyl groups, and triarylsilyl
groups; and other heteroatoms in various other groups.
"Substituted" also means any of the above groups in which one or
more hydrogen atoms are replaced by a higher-order bond (e.g., a
double- or triple-bond) to a heteroatom such as oxygen in oxo,
carbonyl, carboxyl, and ester groups; and nitrogen in groups such
as imines, oximes, hydrazones, and nitriles. For example,
"substituted" includes any of the above groups in which one or more
hydrogen atoms are replaced with --NR.sub.gR.sub.h,
--NR.sub.gC(.dbd.O)R.sub.h, --NR.sub.gC(.dbd.O)NR.sub.gR.sub.h,
--NR.sub.gC(.dbd.O)OR.sub.h, --NR.sub.gSO.sub.2R.sub.h,
--OC(.dbd.O)NR.sub.gR.sub.h, --OR.sub.g, --SR.sub.g, --SOR.sub.g,
--SO.sub.2R.sub.g, --OSO.sub.2R.sub.g, --SO.sub.2OR.sub.g,
.dbd.NSO.sub.2R.sub.g, and --SO.sub.2NR.sub.gR.sub.h. "Substituted"
also means any of the above groups in which one or more hydrogen
atoms are replaced with --C(.dbd.O)R.sub.g, --C(.dbd.O)OR.sub.g,
--C(.dbd.O)NR.sub.gR.sub.h, --CH.sub.2SO.sub.2R.sub.g,
--CH.sub.2SO.sub.2NR.sub.gR.sub.h,
--(CH.sub.2CH.sub.2O).sub.2-10R.sub.g. In the foregoing, R.sub.g
and R.sub.h are the same or different and independently hydrogen,
alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl,
cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl,
heterocyclylalkyl, heteroaryl, N-heteroaryl and/or heteroarylalkyl.
"Substituted" further means any of the above groups in which one or
more hydrogen atoms are replaced by a bond to an amino, cyano,
hydroxyl, imino, nitro, oxo, thioxo, halo, alkyl, alkoxy,
alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl,
haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl,
heteroaryl, N-heteroaryl and/or heteroarylalkyl group. In addition,
each of the foregoing substituents may also be optionally
substituted with one or more of the above substituents.
[0176] "Prodrug" is meant to indicate a compound that may be
converted under physiological conditions or by solvolysis to a
biologically active compound of the invention. Thus, the term
"prodrug" refers to a metabolic precursor of a compound of the
invention that is pharmaceutically acceptable. A prodrug may be
inactive when administered to a subject in need thereof, but is
converted in vivo to an active compound of the invention. Prodrugs
are typically rapidly transformed in vivo to yield the parent
compound of the invention, for example, by hydrolysis in blood. The
prodrug compound often offers advantages of solubility, tissue
compatibility or delayed release in a mammalian organism (see,
Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier,
Amsterdam)). A discussion of prodrugs is provided in Higuchi, T.,
et al., A.C.S. Symposium Series, Vol. 14, and in Bioreversible
Carriers in Drug Design, Ed. Edward B. Roche, American
Pharmaceutical Association and Pergamon Press, 1987.
[0177] The term "prodrug" is also meant to include any covalently
bonded carriers, which release the active compound of the invention
in vivo when such prodrug is administered to a mammalian subject.
Prodrugs of a compound of the invention may be prepared by
modifying functional groups present in the compound of the
invention in such a way that the modifications are cleaved, either
in routine manipulation or in vivo, to the parent compound of the
invention. Prodrugs include compounds of the invention wherein a
hydroxy, amino or mercapto group is bonded to any group that, when
the prodrug of the compound of the invention is administered to a
mammalian subject, cleaves to form a free hydroxy, free amino or
free mercapto group, respectively. Examples of prodrugs include,
but are not limited to, acetate, formate and benzoate derivatives
of alcohol or amide derivatives of amine functional groups in the
compounds of the invention and the like.
[0178] The invention disclosed herein is also meant to encompass
the in vivo metabolic products of the disclosed compounds. Such
products may result from, for example, the oxidation, reduction,
hydrolysis, amidation, esterification, and the like of the
administered compound, primarily due to enzymatic processes.
Accordingly, the invention includes compounds produced by a process
comprising administering a compound of this invention to a mammal
for a period of time sufficient to yield a metabolic product
thereof. Such products are typically identified by administering a
radiolabelled compound of the invention in a detectable dose to an
animal, such as rat, mouse, guinea pig, monkey, or to human,
allowing sufficient time for metabolism to occur, and isolating its
conversion products from the urine, blood or other biological
samples.
[0179] "Stable compound" and "stable structure" are meant to
indicate a compound that is sufficiently robust to survive
isolation to a useful degree of purity from a reaction mixture, and
formulation into an efficacious therapeutic agent.
[0180] "Optional" or "optionally" means that the subsequently
described event or circumstances may or may not occur, and that the
description includes instances where said event or circumstance
occurs and instances in which it does not. For example, "optionally
substituted aryl" means that the aryl radical may or may not be
substituted and that the description includes both substituted aryl
radicals and aryl radicals having no substitution.
[0181] "Pharmaceutically acceptable carrier, diluent or excipient"
includes without limitation any adjuvant, carrier, excipient,
glidant, sweetening agent, diluent, preservative, dye/colorant,
flavor enhancer, surfactant, wetting agent, dispersing agent,
suspending agent, stabilizer, isotonic agent, solvent, or
emulsifier which has been approved by the United States Food and
Drug Administration as being acceptable for use in humans or
domestic animals.
[0182] "Pharmaceutically acceptable salt" includes both acid and
base addition salts.
[0183] "Pharmaceutically acceptable acid addition salt" refers to
those salts which retain the biological effectiveness and
properties of the free bases, which are not biologically or
otherwise undesirable, and which are formed with inorganic acids
such as, but are not limited to, hydrochloric acid, hydrobromic
acid, sulfuric acid, nitric acid, phosphoric acid and the like, and
organic acids such as, but not limited to, acetic acid,
2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid,
aspartic acid, benzenesulfonic acid, benzoic acid,
4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid,
capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic
acid, citric acid, cyclamic acid, dodecylsulfuric acid,
ethane-1,2-disulfonic acid, ethanesulfonic acid,
2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric
acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic
acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid,
glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric
acid, lactic acid, lactobionic acid, lauric acid, maleic acid,
malic acid, malonic acid, mandelic acid, methanesulfonic acid,
mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic
acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid,
orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic
acid, pyroglutamic acid, pyruvic acid, salicylic acid,
4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid,
tartaric acid, thiocyanic acid, p-toluenesulfonic acid,
trifluoroacetic acid, undecylenic acid, and the like.
[0184] "Pharmaceutically acceptable base addition salt" refers to
those salts which retain the biological effectiveness and
properties of the free acids, which are not biologically or
otherwise undesirable. These salts are prepared from addition of an
inorganic base or an organic base to the free acid. Salts derived
from inorganic bases include, but are not limited to, the sodium,
potassium, lithium, ammonium, calcium, magnesium, iron, zinc,
copper, manganese, aluminum salts and the like. Preferred inorganic
salts are the ammonium, sodium, potassium, calcium, and magnesium
salts. Salts derived from organic bases include, but are not
limited to, salts of primary, secondary, and tertiary amines,
substituted amines including naturally occurring substituted
amines, cyclic amines and basic ion exchange resins, such as
ammonia, isopropylamine, trimethylamine, diethylamine,
triethylamine, tripropylamine, diethanolamine, ethanolamine,
deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol,
dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,
hydrabamine, choline, betaine, benethamine, benzathine,
ethylenediamine, glucosamine, methylglucamine, theobromine,
triethanolamine, tromethamine, purines, piperazine, piperidine,
N-ethylpiperidine, polyamine resins and the like. Particularly
preferred organic bases are isopropylamine, diethylamine,
ethanolamine, trimethylamine, dicyclohexylamine, choline and
caffeine.
[0185] Often crystallizations produce a solvate of the compound of
the invention. As used herein, the term "solvate" refers to an
aggregate that comprises one or more molecules of a compound of the
invention with one or more molecules of solvent. The solvent may be
water, in which case the solvate may be a hydrate. Alternatively,
the solvent may be an organic solvent. Thus, the compounds of the
present invention may exist as a hydrate, including a monohydrate,
dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate and
the like, as well as the corresponding solvated forms. The compound
of the invention may be true solvates, while in other cases, the
compound of the invention may merely retain adventitious water or
be a mixture of water plus some adventitious solvent.
[0186] A "pharmaceutical composition" refers to a formulation of a
compound of the invention and a medium generally accepted in the
art for the delivery of the biologically active compound to
mammals, e.g., humans. Such a medium includes all pharmaceutically
acceptable carriers, diluents or excipients therefor.
[0187] The compounds of the invention, or their pharmaceutically
acceptable salts may contain one or more asymmetric centers and may
thus give rise to enantiomers, diastereomers, and other
stereoisomeric forms that may be defined, in terms of absolute
stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino
acids. The present invention is meant to include all such possible
isomers, as well as their racemic and optically pure forms.
Optically active (+) and (-), (R)- and (S)-, or (D)- and
(L)-isomers may be prepared using chiral synthons or chiral
reagents, or resolved using conventional techniques, for example,
chromatography and fractional crystallization. Conventional
techniques for the preparation/isolation of individual enantiomers
include chiral synthesis from a suitable optically pure precursor
or resolution of the racemate (or the racemate of a salt or
derivative) using, for example, chiral high pressure liquid
chromatography (HPLC). When the compounds described herein contain
olefinic double bonds or other centres of geometric asymmetry, and
unless specified otherwise, it is intended that the compounds
include both E and Z geometric isomers. Likewise, all tautomeric
forms are also intended to be included.
[0188] A "stereoisomer" refers to a compound made up of the same
atoms bonded by the same bonds but having different
three-dimensional structures, which are not interchangeable. The
present invention contemplates various stereoisomers and mixtures
thereof and includes "enantiomers", which refers to two
stereoisomers whose molecules are nonsuperimposeable mirror images
of one another.
[0189] A "tautomer" refers to a proton shift from one atom of a
molecule to another atom of the same molecule. The present
invention includes tautomers of any said compounds.
[0190] In accordance with the present disclosure, the compounds
described herein are designed to be substantially active or
localized in the gastrointestinal lumen of a human or animal
subject. The term "gastrointestinal lumen" is used interchangeably
herein with the term "lumen," to refer to the space or cavity
within a gastrointestinal tract (GI tract, which can also be
referred to as the gut), delimited by the apical membrane of GI
epithelial cells of the subject. In some embodiments, the compounds
are not absorbed through the layer of epithelial cells of the GI
tract (also known as the GI epithelium). "Gastrointestinal mucosa"
refers to the layer(s) of cells separating the gastrointestinal
lumen from the rest of the body and includes gastric and intestinal
mucosa, such as the mucosa of the small intestine. A
"gastrointestinal epithelial cell" or a "gut epithelial cell" as
used herein refers to any epithelial cell on the surface of the
gastrointestinal mucosa that faces the lumen of the
gastrointestinal tract, including, for example, an epithelial cell
of the stomach, an intestinal epithelial cell, a colonic epithelial
cell, and the like.
[0191] "Substantially systemically non-bioavailable" and/or
"substantially impermeable" as used herein (as well as variations
thereof) generally refer to situations in which a statistically
significant amount, and in some embodiments essentially all of the
compound of the present disclosure (which includes the
NHE-inhibitor small molecule), remains in the gastrointestinal
lumen. For example, in accordance with one or more embodiments of
the present disclosure, preferably at least about 70%, about 80%,
about 90%, about 95%, about 98%, about 99%, or even about 99.5%, of
the compound remains in the gastrointestinal lumen. In such cases,
localization to the gastrointestinal lumen refers to reducing net
movement across a gastrointestinal layer of epithelial cells, for
example, by way of both transcellular and paracellular transport,
as well as by active and/or passive transport. The compound in such
embodiments is hindered from net permeation of a layer of
gastrointestinal epithelial cells in transcellular transport, for
example, through an apical membrane of an epithelial cell of the
small intestine. The compound in these embodiments is also hindered
from net permeation through the "tight junctions" in paracellular
transport between gastrointestinal epithelial cells lining the
lumen.
[0192] In this regard it is to be noted that, in one particular
embodiment, the compound is essentially not absorbed at all by the
GI tract or gastrointestinal lumen. As used herein, the terms
"substantially impermeable" or "substantially systemically
non-bioavailable" refers to embodiments wherein no detectable
amount of absorption or permeation or systemic exposure of the
compound is detected, using means generally known in the art.
[0193] In this regard it is to be further noted, however, that in
alternative embodiments "substantially impermeable" or
"substantially systemically non-bioavailable" provides or allows
for some limited absorption in the GI tract, and more particularly
the gut epithelium, to occur (e.g., some detectable amount of
absorption, such as for example at least about 0.1%, 0.5%, 1% or
more and less than about 30%, 20%, 10%, 5%, etc., the range of
absorption being for example between about 1% and 30%, or 5% and
20%, etc.; stated another way, "substantially impermeable" or
"substantially systemically non-bioavailable" refers to compounds
that exhibit some detectable permeability to an epithelium layer of
cells in the GI tract of less than about 20% of the administered
compound (e.g., less than about 15%, about 10%, or even about 5%,
and for example greater than about 0.5%, or 1%), but then are
cleared by the liver (i.e., hepatic extraction) and/or the kidney
(i.e., renal excretion).
B. Permeability
[0194] In this regard it is to be noted that, in various
embodiments, the ability of a compound to be substantially
systemically non-bioavailable is based on the compound charge,
size, and/or other physicochemical parameters (e.g., polar surface
area, number of hydrogen bond donors and/or acceptors therein,
number of freely rotatable bonds, etc.). More specifically, it is
to be noted that the absorption character of a compound can be
selected by applying principles of pharmacodynamics, for example,
by applying Lipinski's rule, also known as "the rule of five."
Although not a rule, but rather a set of guidelines, Lipinski shows
that small molecule drugs with (i) a molecular weight, (ii) a
number of hydrogen bond donors, (iii) a number of hydrogen bond
acceptors, and/or (iv) a water/octanol partition coefficient
(Moriguchi Log P), greater than a certain threshold value,
generally do not show significant systemic concentration (i.e., are
generally not absorbed to any significant degree). (See, e.g.,
Lipinski et al., Advanced Drug Delivery Reviews, 46, 2001 3-26,
incorporated herein by reference.) Accordingly, substantially
systemically non-bioavailable compounds (e.g., substantially
systemically non-bioavailable NHE-inhibiting compounds) can be
designed to have molecular structures exceeding one or more of
Lipinski's threshold values. (See also Lipinski et al.,
Experimental and Computational Approaches to Estimate Solubility
and Permeability in Drug Discovery and Development Settings, Adv.
Drug Delivery Reviews, 46:3-26 (2001); and Lipinski, Drug-like
Properties and the Causes of Poor Solubility and Poor Permeability,
J. Pharm. & Toxicol. Methods, 44:235-249 (2000), incorporated
herein by reference.) In some embodiments, for example, a
substantially impermeable or substantially systemically
non-bioavailable NHE-inhibiting compound of the present disclosure
can be constructed to feature one or more of the following
characteristics: (i) a MW greater than about 500 Da, about 1000 Da,
about 2500 Da, about 5000 Da, about 10,000 Da or more (in the
non-salt form of the compound); (ii) a total number of NH and/or OH
and/or other potential hydrogen bond donors greater than about 5,
about 10, about 15 or more; (iii) a total number of 0 atoms and/or
N atoms and/or other potential hydrogen bond acceptors greater than
about 5, about 10, about 15 or more; and/or (iv) a Moriguchi
partition coefficient greater than about 10.sup.5 (i.e., Log P
greater than about 5, about 6, about 7, etc.), or alternatively
less than about 10 (i.e., a Log P of less than 1, or even 0).
[0195] In addition to the parameters noted above, the molecular
polar surface area (i.e., "PSA"), which may be characterized as the
surface belonging to polar atoms, is a descriptor that has also
been shown to correlate well with passive transport through
membranes and, therefore, allows prediction of transport properties
of drugs. It has been successfully applied for the prediction of
intestinal absorption and Caco2 cell monolayer penetration. (For
Caco2 cell monolayer penetration test details, see for example the
description of the Caco2 Model provided in Example 31 of U.S. Pat.
No. 6,737,423, the entire contents of which are incorporated herein
by reference for all relevant and consistent purposes, and the text
of Example 31 in particular, which may be applied for example to
the evaluation or testing of the compounds of the present
disclosure.) PSA is expressed in .ANG..sup.2 (squared angstroms)
and is computed from a three-dimensional molecular representation.
A fast calculation method is now available (see, e.g., Ertl et al.,
Journal of Medicinal Chemistry, 2000, 43, 3714-3717, the entire
contents of which are incorporated herein by reference for all
relevant and consistent purposes) using a desktop computer and
commercially available chemical graphic tools packages, such as
ChemDraw. The term "topological PSA" (tPSA) has been coined for
this fast-calculation method. tPSA is well correlated with human
absorption data with common drugs (see, e.g., Table 1, below):
TABLE-US-00001 TABLE 1 name % FA.sup.a TPSA.sup.b metoprolol 102
50.7 nordiazepam 99 41.5 diazepam 97 32.7 oxprenolol 97 50.7
phenazone 97 26.9 oxazepam 97 61.7 alprenolol 96 41.9 practolol 95
70.6 pindolol 92 57.3 ciprofloxacin 69 74.6 metolazone 64 92.5
tranexamic acid 55 63.3 atenolol 54 84.6 sulpiride 36 101.7
mannitol 26 121.4 foscarnet 17 94.8 sulfasalazine 12 141.3
olsalazine 2.3 139.8 lactilose 0.6 197.4 raffinose 0.3 268.7
(from Ertl et al., J. Med. Chem., 2000, 43:3714-3717). Accordingly,
in some preferred embodiments, the compounds of the present
disclosure may be constructed to exhibit a tPSA value greater than
about 100 .ANG..sup.2, about 120 .ANG..sup.2, about 130
.ANG..sup.2, or about 140 .ANG..sup.2, and in some instances about
150 .ANG..sup.2, about 200 .ANG..sup.2, about 250 .ANG..sup.2,
about 270 .ANG..sup.2, about 300 .ANG..sup.2, about 400
.ANG..sup.2, or even about 500 .ANG..sup.2, such that the compounds
are substantially impermeable or substantially systemically
non-bioavailable (as defined elsewhere herein).
[0196] Because there are exceptions to Lipinski's "rule," or the
tPSA model, the permeability properties of the compounds of the
present disclosure may be screened experimentally. The permeability
coefficient can be determined by methods known to those of skill in
the art, including for example by Caco-2 cell permeability assay
and/or using an artificial membrane as a model of a
gastrointestinal epithelial cell. (As previously noted above, see
for example U.S. Pat. No. 6,737,423, Example 31 for a description
of the Caco-2 Model, which is incorporated herein by reference). A
synthetic membrane impregnated with, for example, lecithin and/or
dodecane to mimic the net permeability characteristics of a
gastrointestinal mucosa, may be utilized as a model of a
gastrointestinal mucosa. The membrane can be used to separate a
compartment containing the compound of the present disclosure from
a compartment where the rate of permeation will be monitored. Also,
parallel artificial membrane permeability assays (PAMPA) can be
performed. Such in vitro measurements can reasonably indicate
actual permeability in vivo. (See, for example, Wohnsland et al.,
J. Med. Chem., 2001, 44:923-930; Schmidt et al., Millipore Corp.
Application Note, 2002, n.sup.o AN1725EN00, and n.sup.o AN1728EN00,
incorporated herein by reference.)
[0197] Accordingly, in some embodiments, the compounds utilized in
the methods of the present disclosure may have a permeability
coefficient, P.sub.app, of less than about 100.times.10.sup.-6
cm/s, or less than about 10.times.10.sup.-6 cm/s, or less than
about 1.times.10.sup.-6 cm/s, or less than about
0.1.times.10.sup.-6 cm/s, when measured using means known in the
art (such as for example the permeability experiment described in
Wohnsland et al., J. Med. Chem., 2001, 44. 923-930, the contents of
which is incorporated herein by reference).
[0198] As previously noted, in accordance with the present
disclosure, a NHE-inhibiting small molecule moiety is modified as
described above to hinder the net absorption through a layer of gut
epithelial cells, rendering the resulting compound substantially
systemically non-bioavailable. In various embodiments, the
compounds of the present disclosure comprise an NHE-inhibiting
small molecule moiety linked, coupled or otherwise attached to a
moiety which renders the overall compound substantially impermeable
or substantially systemically non-bioavailable. More specifically,
the NHE-inhibiting small molecule moiety is coupled to a dimer,
multimer or polymer moiety, such that the resulting compound is
substantially impermeable or substantially systemically
non-bioavailable. The dimer, multimer or polymer portion or moiety
may be of a molecular weight greater than about 500 Daltons (Da),
about 1000 Da, about 2500 Da, about 5000 Da, about 10,000 Da or
more, and in particular may have a molecular weight in the range of
about 1000 Daltons (Da) to about 500,000 Da, preferably in the
range of about 5000 to about 200,000 Da, and more preferably may
have a molecular weight that is sufficiently high to essentially
preclude any net absorption through a layer of gut epithelial cells
of the compound.
C. Persistent Inhibitory Effect
[0199] In other embodiments, the substantially impermeable or
substantially systemically non-bioavailable NHE-inhibiting
compounds utilized in the treatment methods of the present
disclosure may additionally exhibit a persistent inhibitor effect.
This effect manifests itself when the inhibitory action of a
compound at a certain concentration in equilibrium with the
epithelial cell (e.g., at or above its inhibitory concentration,
IC) does not revert to baseline (i.e., sodium transport without
inhibitor) after the compound is depleted by simple washing of the
luminal content.
[0200] This effect can be interpreted as a result of the tight
binding of the NHE-inhibiting compounds to the NHE protein at the
intestinal apical side of the gut epithelial cell. The binding can
be considered as quasi-irreversible to the extent that, after the
compound has been contacted with the gut epithelial cell and
subsequently washed off said gut epithelial cell, the flux of
sodium transport is still significantly lower than in the control
without the compound. This persistent inhibitory effect has the
clear advantage of maintaining drug activity within the GI tract
even though the residence time of the active in the upper GI tract
is short, and when no entero-biliary recycling process is effective
to replenish the compound concentration near its site of
action.
[0201] Such a persistent inhibitory effect has an obvious advantage
in terms of patient compliance, but also in limiting drug exposure
within the GI tract.
[0202] The persistence effect can be determined using in vitro
methods; in one instance, cell lines expressing NHE transporters
are split in different vials and treated with a NHE-inhibiting
compound and sodium solution to measure the rate of sodium uptake.
The cells in one set of vials are washed for different periods of
time to remove the inhibitor, and sodium uptake measurement is
repeated after the washing. Compounds that maintain their
inhibitory effect after multiple/lengthy washing steps (compared to
the inhibitory effect measured in the vials where washing does not
occur) are persistent inhibitors. Persistence effect can also be
characterized ex vivo by using the everted sac technique, whereby
transport of Na is monitored using an excised segment of GI
perfused with a solution containing the inhibitor and shortly after
flushing the bathing solution with a buffer solution free from
inhibitor. A persistence effect can also be characterized in vivo
by observing the time needed for sodium balance to return to normal
when the inhibitor treatment is discontinued. The limit of the
method resides in the fact that apical cells (and therefore apical
NHE transporters) are sloughed off after a period of 3 to 4 days,
the typical turnover time of gut epithelial cells. A persistence
effect can be achieved by increasing the residence time of the
active compound at the apical surface of the gut epithelial cells;
this can be obtained by designing NHE antiport inhibitors with
several NHE-inhibiting small molecule moieties built-in the small
molecule or oligomer (wherein "several" as used herein typically
means at least about 2, about 4, about 6 or more). Examples of such
structures in the context of analogs of the antibiotic vancomycin
are given in Griffin, et al., J. Am. Chem. Soc., 2003, 125,
6517-6531. Alternatively the compound comprises groups that
contribute to increase the affinity towards the gut epithelial cell
so as to increase the time of contact with the gut epithelial cell
surface. Such groups are referred to as being "mucoadhesive." More
specifically, the Core or L moiety can be substituted by such
mucoadhesive groups, such as polyacrylates, partially deacetylated
chitosan or polyalkylene glycol. (See also Patil, S. B. et al.,
Curr. Drug. Deliv., 2008, Oct. 5(4), pp. 312-8.)
D. GI Enzyme Resistance
[0203] Because the compounds utilized in the treatment methods of
the present disclosure are preferably substantially systemically
non-bioavailable, and/or preferably exhibit a persistent inhibitory
effect, it is also desirable that, during their prolonged residence
time in the gut, these compounds sustain the hydrolytic conditions
prevailing in the upper GI tract. In such embodiments, compounds of
the present disclosure are resistant to enzymatic metabolism. For
example, administered compounds are preferably resistant to the
activity of P450 enzymes, glucurosyl transferases,
sulfotransferases, glutathione S-transferases, and the like, in the
intestinal mucosa, as well as gastric (e.g., gastric lipase, and
pepsine), pancreatic (e.g., trypsin, triglyceride pancreatic
lipase, phospholipase A2, endonucleases, nucleotidases, and
alpha-amylase), and brush-border enzymes (e.g., alkaline
phosphatase, glycosidases, and proteases) generally known in the
art.
[0204] The compounds that are utilized in methods of the present
disclosure are also preferably resistant to metabolism by the
bacterial flora of the gut; that is, the compounds are not
substrates for enzymes produced by bacterial flora. In addition,
the compounds administered in accordance with the methods of the
present disclosure may be substantially inactive towards the
gastrointestinal flora, and do not disrupt bacterial growth or
survival. As a result, in various embodiments herein, the minimal
inhibitory concentration (or "MIC") against GI flora is desirably
greater than about 15 .mu.g/ml, about 30 .mu.g/ml, about 60
.mu.g/ml, about 120 .mu.g/ml, or even about 240 .mu.g/ml, the MIC
in various embodiments being for example between about 16 and about
32 .mu.g/ml, or between about 64 and about 128 .mu.g/ml, or greater
than about 256 .mu.g/ml.
[0205] To one skilled in the art of medicinal chemistry, metabolic
stability can be achieved in a number of ways. Functionality
susceptible to P450-mediated oxidation can be protected by, for
example, blocking the point of metabolism with a halogen or other
functional group. Alternatively, electron withdrawing groups can be
added to a conjugated system to generally provide protection to
oxidation by reducing the electrophilicity of the compound.
Proteolytic stability can be achieved by avoiding secondary amide
bonds, or by incorporating changes in stereochemistry or other
modifications that prevent the drug from otherwise being recognized
as a substrate by the metabolizing enzyme.
E. Sodium and/or Fluid Output
[0206] It is also to be noted that, in various embodiments of the
present disclosure, one or more of the NHE-inhibiting compounds
detailed herein, when administered either alone or in combination
with one or more additional pharmaceutically active compounds or
agents (including, for example, a fluid-absorbing polymer) to a
patient in need thereof, may act to increase the patient's daily
fecal output of sodium by at least about 20, about 30 mmol, about
40 mmol, about 50 mmol, about 60 mmol, about 70 mmol, about 80
mmol, about 90 mmol, about 100 mmol, about 125 mmol, about 150 mmol
or more, the increase being for example within the range of from
about 20 to about 150 mmol/day, or from about 25 to about 100
mmol/day, or from about 30 to about 60 mmol/day
[0207] Additionally, or alternatively, it is also to be noted that,
in various embodiments of the present disclosure, one or more of
the NHE-inhibiting compounds detailed herein, when administered
either alone or in combination with one or more additional
pharmaceutically active compounds or agents (including, for
example, a fluid-absorbing polymer) to a patent in need thereof,
may act to increase the patient's daily fluid output by at least
about 100 ml, about 200 ml, about 300 ml, about 400 ml, about 500
ml, about 600 ml, about 700 ml, about 800 ml, about 900 ml, about
1000 ml or more, the increase being for example within the range of
from about 100 to about 1000 ml/day, or from about 150 to about 750
ml/day, or from about 200 to about 500 ml/day (assuming isotonic
fluid).
F. C.sub.max and IC.sub.50
[0208] It is also to be noted that, in various embodiments of the
present disclosure, one or more of the NHE-inhibiting compounds
detailed herein, when administered either alone or in combination
with one or more additional pharmaceutically active compounds or
agents (including, for example, a fluid-absorbing polymer) to a
patient in need thereof at a dose resulting in at least a 10%
increase in fecal water content, has a C.sub.max that is less than
the IC.sub.50 for NHE-3, more specifically, less than about
10.times. (10 times) the IC.sub.50, and, more specifically still,
less than about 100.times. (100 times) the IC.sub.50.
[0209] Additionally, or alternatively, it is also to be noted that,
in various embodiments of the present disclosure, one or more of
the NHE-inhibiting compounds detailed herein, when administered
either alone or in combination with one or more additional
pharmaceutically active compounds or agents (including, for
example, a fluid-absorbing polymer) to a patient in need thereof,
may have a C.sub.max of less than about 10 ng/ml, about 7.5 ng/ml,
about 5 ng/ml, about 2.5 ng/ml, about 1 ng/ml, or about 0.5 ng/ml,
the C.sub.max being for example within the range of about 1 ng/ml
to about 10 ng/ml, or about 2.5 ng/ml to about 7.5 ng/ml.
[0210] Additionally, or alternatively, it is also to be noted that,
in various embodiments of the present disclosure, one or more of
the NHE-inhibiting compounds detailed herein, when administered
either alone or in combination with one or more additional
pharmaceutically active compounds or agents (including, for
example, a fluid-absorbing polymer) to a patient in need thereof,
may have a IC.sub.50 of less than about 10 .mu.M, about 7.5 .mu.M,
about 5 .mu.M, about 2.5 .mu.M, about 1 .mu.M, or about 0.5 .mu.M,
the IC.sub.50 being for example within the range of about 1 .mu.M
to about 10 .mu.M, or about 2.5 .mu.M to about 7.5 .mu.M.
[0211] Additionally, or alternatively, it is also to be noted that,
in various embodiments of the present disclosure, one or more of
the NHE-inhibiting compounds detailed herein, when administered to
a patient in need thereof, may have a ratio of IC.sub.50:C.sub.max,
wherein IC.sub.50 and C. are expressed in terms of the same units,
of at least about 10, about 50, about 100, about 250, about 500,
about 750, or about 1000.
[0212] Additionally, or alternatively, it is also to be noted that,
in various embodiments of the present disclosure, wherein one or
more of the NHE-inhibiting compounds as detailed herein is orally
administered to a patent in need thereof, within the therapeutic
range or concentration, the maximum compound concentration detected
in the serum, defined as C., is lower than the NHE inhibitory
concentration IC.sub.50 of said compound. As previously noted, as
used herein, IC.sub.50 is defined as the quantitative measure
indicating the concentration of the compound required to inhibit
50% of the NHE-mediated Na/H antiport activity in a cell based
assay.
III. Pharmaceutical Compositions and Methods of Treatment
A. Compositions and Methods
1. Fluid Retention and/or Salt Overload Disorders
[0213] A pharmaceutical composition or preparation that may be used
in accordance with the present disclosure for the treatment of
various disorders associated with fluid retention and/or salt
overload in the gastrointestinal tract (e.g., hypertension, heart
failure (in particular, congestive heart failure), chronic kidney
disease, end-stage renal disease, liver disease and/or peroxisome
proliferator-activated receptor (PPAR) gamma agonist-induced fluid
retention) comprises, in general, the substantially impermeable or
substantially systemically non-bioavailable NHE-inhibiting compound
of the present disclosure, as well as various other optional
components as further detailed herein below (e.g., pharmaceutically
acceptable excipients, etc.). The compounds utilized in the
treatment methods of the present disclosure, as well as the
pharmaceutical compositions comprising them, may accordingly be
administered alone, or as part of a treatment protocol or regiment
that includes the administration or use of other beneficial
compounds (as further detailed elsewhere herein). In some
particular embodiments, the NHE-inhibiting compound, including any
pharmaceutical composition comprising the compound, is administered
with a fluid-absorbing polymer (as more fully described below).
[0214] A "subject" or "mammal" is preferably a human, but can also
be an animal in need of treatment with a compound of the
disclosure, e.g., companion animals (e.g., dogs, cats, and the
like), farm animals (e.g., cows, pigs, horses and the like) and
laboratory animals (e.g., rats, mice, guinea pigs and the
like).
[0215] Subjects "in need of treatment" with a compound of the
present disclosure, or subjects "in need of NHE inhibition" include
subjects with diseases and/or conditions that can be treated with
substantially impermeable or substantially systemically
non-bioavailable NHE-inhibiting compounds, with or without a
fluid-absorbing polymer, to achieve a beneficial therapeutic and/or
prophylactic result. A beneficial outcome includes a decrease in
the severity of symptoms or delay in the onset of symptoms,
increased longevity and/or more rapid or more complete resolution
of the disease or condition. For example, a subject in need of
treatment may be suffering from hypertension; from salt-sensitive
hypertension which may result from dietary salt intake; from a risk
of a cardiovascular disorder (e.g., myocardial infarction,
congestive heart failure and the like) resulting from hypertension;
from heart failure (e.g., congestive heart failure) resulting in
fluid or salt overload; from chronic kidney disease resulting in
fluid or salt overload, from end stage renal disease resulting in
fluid or salt overload; from liver disease resulting in fluid or
salt overload; from peroxisome proliferator-activated receptor
(PPAR) gamma agonist-induced fluid retention; or from edema
resulting from congestive heart failure or end stage renal disease.
In various embodiments, a subject in need of treatment typically
shows signs of hypervolemia resulting from salt and fluid retention
that are common features of congestive heart failure, renal failure
or liver cirrhosis. Fluid retention and salt retention manifest
themselves by the occurrence of shortness of breath, edema, ascites
or interdialytic weight gain. Other examples of subjects that would
benefit from the treatment are those suffering from congestive
heart failure and hypertensive patients and, particularly, those
who are resistant to treatment with diuretics, i.e., patients for
whom very few therapeutic options are available. A subject "in need
of treatment" also includes a subject with hypertension,
salt-sensitive blood pressure and subjects with systolic/diastolic
blood pressure greater than about 130-139/85-89 mm Hg.
[0216] Administration of NHE-inhibiting compounds, with or without
administration of fluid-absorbing polymers, may be beneficial for
patients put on "non-added salt" dietary regimen (i.e., 60-100 mmol
of Na per day), to liberalize their diet while keeping a neutral or
slightly negative sodium balance (i.e., the overall uptake of salt
would be equal of less than the secreted salt). In that context,
"liberalize their diet" means that patients treated may add salt to
their meals to make the meals more palatable, or/and diversify
their diet with salt-containing foods, thus maintaining a good
nutritional status while improving their quality of life.
[0217] The treatment methods described herein may also help
patients with edema associated with chemotherapy, pre-menstrual
fluid overload and preeclampsia (pregnancy-induced
hypertension).
[0218] Accordingly, it is to be noted that the present disclosure
is further directed to methods of treatment involving the
administration of the compound of the present disclosure, or a
pharmaceutical composition comprising such a compound. Such methods
may include, for example, a method for treating hypertension, the
method comprising administering to the patient a substantially
impermeable or substantially systemically non-bioavailable
NHE-inhibiting compound, or a pharmaceutical composition comprising
it. The method may be for reducing fluid overload associated with
heart failure (in particular, congestive heart failure), the method
comprising administering to the patient a substantially impermeable
or substantially systemically non-bioavailable NHE-inhibiting
compound or pharmaceutical composition comprising it. The method
may be for reducing fluid overload associated with end stage renal
disease, the method comprising administering to the patient a
substantially impermeable or substantially systemically
non-bioavailable NHE-inhibiting compound or composition comprising
it. The method may be for reducing fluid overload associated with
peroxisome proliferator-activated receptor (PPAR) gamma agonist
therapy, the method comprising administering to the patient a
substantially impermeable or substantially systemically
non-bioavailable NHE-inhibiting compound or composition comprising
it. Additionally, or alternatively, the method may be for
decreasing the activity of an intestinal NHE transporter in a
patient, the method comprising: administering to the patient a
substantially impermeable or substantially systemically
non-bioavailable NHE-inhibiting compound, or a composition
comprising it.
2. Gastrointestinal Tract Disorders
[0219] A pharmaceutical composition or preparation that may be used
in accordance with the present disclosure for the treatment of
various gastrointestinal tract disorders, including the treatment
or reduction of pain associated with gastrointestinal tract
disorders, comprises, the substantially impermeable or
substantially systemically non-bioavailable NHE-inhibiting compound
of the present disclosure, as well as various other optional
components as further detailed herein below (e.g., pharmaceutically
acceptable excipients, etc.). The compounds utilized in the
treatment methods of the present disclosure, as well as the
pharmaceutical compositions comprising them, may accordingly be
administered alone, or as part of a treatment protocol or regiment
that includes the administration or use of other beneficial
compounds (as further detailed elsewhere herein). In some
particular embodiments, the NHE-inhibiting compound, including any
pharmaceutical composition comprising the compound, is administered
with a fluid-absorbing polymer (as more fully described below).
[0220] A "subject" is preferably a human, but can also be an animal
in need of treatment with a compound of the disclosure, e.g.,
companion animals (e.g., dogs, cats, and the like), farm animals
(e.g., cows, pigs, horses and the like) and laboratory animals
(e.g., rats, mice, guinea pigs and the like).
[0221] Subjects "in need of treatment" with a compound of the
present disclosure, or subjects "in need of NHE inhibition" include
subjects with diseases and/or conditions that can be treated with
substantially impermeable or substantially systemically
non-bioavailable NHE-inhibiting compounds, with or without a
fluid-absorbing polymer, to achieve a beneficial therapeutic and/or
prophylactic result. A beneficial outcome includes a decrease in
the severity of symptoms or delay in the onset of symptoms,
increased longevity and/or more rapid or more complete resolution
of the disease or condition. For example, a subject in need of
treatment is suffering from a gastrointestinal tract disorder; the
patient is suffering from a disorder selected from the group
consisting of: a gastrointestinal motility disorder, irritable
bowel syndrome, chronic constipation, chronic idiopathic
constipation, chronic constipation occurring in cystic fibrosis
patients, chronic constipation occurring in chronic kidney disease
patients, calcium-induced constipation in osteoporotic patients,
opioid-induced constipation, a functional gastrointestinal tract
disorder, gastroesophageal reflux disease, functional heartburn,
dyspepsia, functional dyspepsia, non-ulcer dyspepsia,
gastroparesis, chronic intestinal pseudo-obstruction, Crohn's
disease, ulcerative colitis and related diseases referred to as
inflammatory bowel syndrome, colonic pseudo-obstruction, and the
like.
[0222] In various preferred embodiments, the constipation to be
treated is: associated with the use of a therapeutic agent;
associated with a neuropathic disorder; post-surgical constipation
(postoperative ileus); associated with a gastrointestinal tract
disorder; idiopathic (functional constipation or slow transit
constipation); associated with neuropathic, metabolic or endocrine
disorder (e.g., diabetes mellitus, renal failure, hypothyroidism,
hyperthyroidism, hypocalcaemia, Multiple Sclerosis, Parkinson's
disease, spinal cord lesions, neurofibromatosis, autonomic
neuropathy, Chagas disease, Hirschsprung's disease or cystic
fibrosis, and the like). Constipation may also be the result of
surgery (postoperative ileus) or due the use of drugs such as
analgesics (e.g., opioids), antihypertensives, anticonvulsants,
antidepressants, antispasmodics and antipsychotics.
[0223] Accordingly, it is to be noted that the present disclosure
is further directed to methods of treatment involving the
administration of the compound of the present disclosure, or a
pharmaceutical composition comprising such a compound. Such methods
may include, for example, a method for increasing gastrointestinal
motility in a patient, the method comprising administering to the
patient a substantially non-permeable or substantially
non-bioavailable NHE-inhibiting compound, or a pharmaceutical
composition comprising it. Additionally, or alternatively, the
method may be for decreasing the activity of an intestinal NHE
transporter in a patient, the method comprising administering to
the patient a substantially non-permeable or substantially
non-bioavailable NHE-inhibiting compound, or a pharmaceutical
composition comprising it. Additionally, or alternatively, the
method may be for treating a gastrointestinal tract disorder, a
gastrointestinal motility disorder, irritable bowel syndrome,
chronic calcium-induced constipation in osteoporotic patients,
chronic constipation occurring in cystic fibrosis patients, chronic
constipation occurring in chronic kidney disease patients, a
functional gastrointestinal tract disorder, gastroesophageal reflux
disease, functional heartburn, dyspepsia, functional dyspepsia,
non-ulcer dyspepsia, gastroparesis, chronic intestinal
pseudo-obstruction, colonic pseudo-obstruction, Crohn's disease,
ulcerative colitis, inflammatory bowel disease, the method
comprising administering an antagonist of the intestinal NHE, and
more specifically, a substantially non-permeable or substantially
non-bioavailable NHE-inhibiting compound, or a pharmaceutical
composition comprising it, either orally or by rectal suppository.
Additionally, or alternatively, the method may be for treating or
reducing pain, including visceral pain, pain associated with a
gastrointestinal tract disorder or pain associated with some other
disorder, the method comprising administering to a patient a
substantially non-permeable or substantially non-bioavailable
NHE-inhibiting compound, or a pharmaceutical composition comprising
it. Additionally, or alternatively, the method may be for treating
inflammation, including inflammation of the gastrointestinal tract,
e.g., inflammation associated with a gastrointestinal tract
disorder or infection or some other disorder, the method comprising
administering to a patient a substantially non-permeable or
substantially non-bioavailable NHE-inhibiting compound, or a
pharmaceutical composition comprising it.
3. Metabolic Disorders
[0224] A pharmaceutical composition or preparation that may be used
in accordance with the present disclosure for the treatment of
various metabolic disorders including the treatment or reduction of
type II diabetes mellitus (T2DM), metabolic syndrome, and/or
symptoms associated with such disorders comprises, in general, the
substantially impermeable or substantially systemically
non-bioavailable NHE-inhibiting compound of the present disclosure,
as well as various other optional components as further detailed
herein below (e.g., pharmaceutically acceptable excipients, etc.).
The compounds utilized in the treatment methods of the present
disclosure, as well as the pharmaceutical compositions comprising
them, may accordingly be administered alone, or as part of a
treatment protocol or regiment that includes the administration or
use of other beneficial compounds (as further detailed elsewhere
herein).
[0225] Obesity is becoming a worldwide epidemic. In the United
States, approximately 2/3rds of the population is either overweight
(body mass index [BMI] 25 to 29.9) or obese (BMI .gtoreq.30)
(Ogden, C L et al, "Prevalence of overweight and obesity in the
united states, 1999-2004" JAMA 2006, 295, 1549-1555). Obesity is a
major risk factor for the development of diabetes and related
complications, including cardiovascular disease and chronic kidney
disease (CKD). The prevalence of T2DM has increased alarmingly in
the United States. The American Diabetes Associated (ADA) estimates
that more than 23 million U.S. adults aged 20 years or older have
diabetes, with T2DM accounting for approximately 95% of these
cases. The World Health Organization (WHO) has put the number of
persons with diabetes worldwide at approximately 170 million
(Campbell, R. K. "Type 2 diabetes: where we are today: an overview
of disease burden, current treatments, and treatment strategies"
Journal of the American Pharmacists Association 2009, 49(5),
S3-S9).
[0226] Obesity is also a major risk factor for the development of
metabolic syndrome, and subsequently the development of CKD.
Metabolic syndrome, previously known as Syndrome X, the
plurimetabolic syndrome, the dysmetabolic syndrome, and other
names, consists of a clustering of metabolic abnormalities
including abdominal obesity, hypertriglyceridemia, low levels of
high-density lipoprotein (HDL) cholesterol, elevated blood pressure
(BP), and elevations in fasting glucose or diabetes (Townsend, R.
R. et al "Metabolic Syndrome, Components, and Cardiovascular
Disease Prevalence in Chronic Kidney Disease: Findings from the
Chronic Renal Insufficiency Cohort (CRIC) Study" American Journal
of Nephrology 2011, 33, 477-484). Metabolic syndrome is common in
patients with CKD and an important risk factor for the development
and progression of CKD.
[0227] Hemodynamic factors appear to play a significant role in
obesity-induced renal dysfunction. Hypertension, which is closely
linked to obesity, appears to be a major cause of renal dysfunction
in obese patients (Wahba, I. M. et al "Obesity and
obesity-initiated metabolic syndrome: mechanistic links to chronic
kidney disease" Clinical Journal of the American Society of
Nephrology 2007, 2, 550-562). Studies in animals and in humans have
shown that obesity is associated with elevated glomerular
filtration rate (GFR) and increased renal blood flow. This likely
occurs because of afferent arteriolar dilation as a result of
proximal salt reabsorption, coupled with efferent renal arteriolar
vasoconstriction as a result of elevated angiotensin II levels.
These effects may contribute to hyperfiltration, glomerulomegaly,
and later focal glomerulosclerosis. Even though GFR is increased in
obesity, urinary sodium excretion in response to a saline load is
often delayed, and individuals exhibit an abnormal pressure
natriuresis, indicating avid proximal tubular sodium reabsorption.
In addition, increased fat distribution can cause increased
intra-abdominal pressure, leading to renal vein compression, thus
raising renal venous pressure and diminishing renal perfusion. In
creased fat, through a variety of mechanisms, can cause elevated
renal interstitial fluid hydrostatic fluid and may stimulate renal
sodium retention the thereby contribute to hypertension (Wahba
2007).
[0228] In view of the above, there exists a need in the art for
agents that can divert sodium and fluid from a subject via
mechanisms that either avoid the kidney, or do not depend upon
normal kidney function. A "subject" with metabolic disease,
including T2DM, metabolic syndrome, and the like, is preferably a
human, but can also be an animal in need of treatment with a
compound of the disclosure, e.g., companion animals (e.g., dogs,
cats, and the like), farm animals (e.g., cows, pigs, horses and the
like) and laboratory animals (e.g., rats, mice, guinea pigs and the
like).
[0229] Subjects "in need of treatment" with a compound of the
present disclosure, or subjects "in need of NHE inhibition" include
subjects with diseases and/or conditions that can be treated with
substantially impermeable or substantially systemically
non-bioavailable NHE-inhibiting compounds, with or without a
fluid-absorbing polymer, to achieve a beneficial therapeutic and/or
prophylactic result. A beneficial outcome includes a decrease in
the severity of symptoms or delay in the onset of symptoms,
increased longevity and/or more rapid or more complete resolution
of the disease or condition. For example, a subject with a
metabolic disorder causing or exacerbating chronic kidney disease
would benefit from a treatment modality that could divert excess
sodium and fluid from the body by a method that does not require
normally functionaling kidneys. Such a treatment would include the
method comprising administering to a patient a substantially
non-permeable or substantially non-bioavailable NHE-inhibiting
compound, or a pharmaceutical composition comprising it.
[0230] The compounds utilized in the treatment methods of the
present disclosure, as well as the pharmaceutical compositions
comprising them, may accordingly be administered alone, or as part
of a combination therapy or regimen that includes the
administration or use of other therapeutic compounds related to the
treatment of metabolic disorders such as T2DM and metabolic
syndrome. In some particular embodiments, the NHE-inhibiting
compound, including any pharmaceutical composition comprising the
compound, is administered with a fluid absorbing polymer.
B. Combination Therapies
1. Fluid Retention and/or Salt Overload Disorders
[0231] As previously noted, the compounds described herein can be
used alone or in combination with other agents. For example, the
compounds can be administered together with a diuretic (i.e., High
Ceiling Loop Diuretics, Benzothiadiazide Diuretics, Potassium
Sparing Diuretics, Osmotic Diuretics), cardiac glycoside, ACE
inhibitor, angiotensin-2 receptor antagonist, calcium channel
blocker, beta blocker, alpha blocker, central alpha agonist,
aldosterone antagonist, aldosterone synthase inhibitor, renin
inhibitor, vasodilator, blood thinner, anti-platelet agent,
lipid-lowering agent, peroxisome proliferator-activated receptor
(PPAR) gamma agonist agent or compound or with a fluid-absorbing
polymer as more fully described below. The agent can be covalently
attached to a compound described herein or it can be a separate
agent that is administered together with or sequentially with a
compound described herein in a combination therapy.
[0232] Combination therapy can be achieved by administering two or
more agents, e.g., a substantially non-permeable or substantially
systemically non-bioavailable NHE-inhibiting compound described
herein and a diuretic, cardiac glycoside, ACE inhibitor,
angiotensin-2 receptor antagonist, aldosterone antagonist,
aldosterone synthase inhibitor, renin inhibitor, calcium channel
blocker, beta blocker, alpha blocker, central alpha agonist,
vasodilator, blood thinner, anti-platelet agent or compound, each
of which is formulated and administered separately, or by
administering two or more agents in a single formulation. Other
combinations are also encompassed by combination therapy. For
example, two agents can be formulated together and administered in
conjunction with a separate formulation containing a third agent.
While the two or more agents in the combination therapy can be
administered simultaneously, they need not be. For example,
administration of a first agent (or combination of agents) can
precede administration of a second agent (or combination of agents)
by minutes, hours, days, or weeks. Thus, the two or more agents can
be administered within minutes of each other or within 1, 2, 3, 6,
9, 12, 15, 18, or 24 hours of each other or within 1, 2, 3, 4, 5,
6, 7, 8, 9, 10, 12, 14 days of each other or within 2, 3, 4, 5, 6,
7, 8, 9, or weeks of each other. In some cases even longer
intervals are possible. While in many cases it is desirable that
the two or more agents used in a combination therapy be present in
within the patient's body at the same time, this need not be
so.
[0233] Combination therapy can also include two or more
administrations of one or more of the agents used in the
combination. For example, if agent X and agent Y are used in a
combination, one could administer them sequentially in any
combination one or more times, e.g., in the order X--Y--X, X--X--Y,
Y--X--Y, Y--Y--X, X--X--Y--Y, etc.
[0234] The compounds described herein can be used in combination
therapy with a diuretic. Among the useful diuretic agents are, for
example: High Ceiling Loop Diuretics [Furosemide (Lasix),
Ethacrynic Acid (Edecrin), Bumetanide (Bumex)], Benzothiadiazide
Diuretics [Hydrochlorothiazide (Hydrodiuril), Chlorothiazide
(Diuril), Clorthalidone (Hygroton), Benzthiazide (Aguapres),
Bendroflumethiazide (Naturetin), Methyclothiazide (Aguatensen),
Polythiazide (Renese), Indapamide (Lozol), Cyclothiazide
(Anhydron), Hydroflumethiazide (Diucardin), Metolazone (Diulo),
Quinethazone (Hydromox), Trichlormethiazide (Naqua)], Potassium
Sparing Diuretics [Spironolactone (Aldactone), Triamterene
(Dyrenium), Amiloride (Midamor)], and Osmotic Diuretics [Mannitol
(Osmitrol)]. Diuretic agents in the various classes are known and
described in the literature.
[0235] Cardiac glycosides (cardenolides) or other digitalis
preparations can be administered with the compounds of the
disclosure in co-therapy. Among the useful cardiac glycosides are,
for example: Digitoxin (Crystodigin), Digoxin (Lanoxin) or
Deslanoside (Cedilanid-D). Cardiac glycosides in the various
classes are described in the literature.
[0236] Angiotensin Converting Enzyme Inhibitors (ACE Inhibitors)
can be administered with the compounds of the disclosure in
co-therapy. Among the useful ACE inhibitors are, for example:
Captopril (Capoten), Enalapril (Vasotec), Lisinopril (Prinivil).
ACE inhibitors in the various classes are described in the
literature.
[0237] Angiotensin-2 Receptor Antagonists (also referred to as
AT.sub.1-antagonists or angiotensin receptor blockers, or ARB's)
can be administered with the compounds of the disclosure in
co-therapy. Among the useful Angiotensin-2 Receptor Antagonists
are, for example: Candesartan (Atacand), Eprosartan (Teveten),
Irbesartan (Avapro), Losartan (Cozaar), Telmisartan (Micardis),
Valsartan (Diovan). Angiotensin-2 Receptor Antagonists in the
various classes are described in the literature.
[0238] Calcium channel blockers such as Amlodipine (Norvasc,
Lotrel), Bepridil (Vascor), Diltiazem (Cardizem, Tiazac),
Felodipine (Plendil), Nifedipine (Adalat, Procardia), Nimodipine
(Nimotop), Nisoldipine (Sular), Verapamil (Calan, Isoptin, Verelan)
and related compounds described in, for example, EP 625162B1, U.S.
Pat. No. 5,364,842, U.S. Pat. No. 5,587,454, U.S. Pat. No.
5,824,645, U.S. Pat. No. 5,859,186, U.S. Pat. No. 5,994,305, U.S.
Pat. No. 6,087,091, U.S. Pat. No. 6,136,786, WO 93/13128 A1, EP
1336409 A1, EP 835126 A1, EP 835126 B1, U.S. Pat. No. 5,795,864,
U.S. Pat. No. 5,891,849, U.S. Pat. No. 6,054,429, WO 97/01351 A1,
the entire contents of which are incorporated herein by reference
for all relevant and consistent purposes, can be used with the
compounds of the disclosure.
[0239] Beta blockers can be administered with the compounds of the
disclosure in co-therapy. Among the useful beta blockers are, for
example: Acebutolol (Sectral), Atenolol (Tenormin), Betaxolol
(Kerlone), Bisoprolol/hydrochlorothiazide (Ziac), Bisoprolol
(Zebeta), Carteolol (Cartrol), Metoprolol (Lopressor, Toprol XL),
Nadolol (Corgard), Propranolol (Inderal), Sotalol (Betapace),
Timolol (Blocadren). Beta blockers in the various classes are
described in the literature.
[0240] PPAR gamma agonists such as thiazolidinediones (also called
glitazones) can be administered with the compounds of the
disclosure in co-therapy. Among the useful PPAR agonists are, for
example: rosiglitazone (Avandia), pioglitazone (Actos) and
rivoglitazone.
[0241] Aldosterone antagonists can be administered with the
compounds of the disclosure in co-therapy. Among the useful
Aldosterone antagonists are, for example: eplerenone,
spironolactone, and canrenone.
[0242] Renin inhibitors can be administered with the compounds of
the disclosure in co-therapy. Among the useful Renin inhibitors is,
for example: aliskiren.
[0243] Alpha blockers can be administered with the compounds of the
disclosure in co-therapy. Among the useful Alpha blockers are, for
example: Doxazosin mesylate (Cardura), Prazosin hydrochloride
(Minipress). Prazosin and polythiazide (Minizide), Terazosin
hydrochloride (Hytrin). Alpha blockers in the various classes are
described in the literature.
[0244] Central alpha agonists can be administered with the
compounds of the disclosure in co-therapy. Among the useful Central
alpha agonists are, for example: Clonidine hydrochloride
(Catapres), Clonidine hydrochloride and chlorthalidone (Clorpres,
Combipres), Guanabenz Acetate (Wytensin), Guanfacine hydrochloride
(Tenex), Methyldopa (Aldomet), Methyldopa and chlorothiazide
(Aldochlor), Methyldopa and hydrochlorothiazide (Aldoril). Central
alpha agonists in the various classes are described in the
literature.
[0245] Vasodilators can be administered with the compounds of the
disclosure in co-therapy. Among the useful vasodilators are, for
example: Isosorbide dinitrate (Isordil), Nesiritide (Natrecor),
Hydralazine (Apresoline), Nitrates/nitroglycerin, Minoxidil
(Loniten). Vasodilators in the various classes are described in the
literature.
[0246] Blood thinners can be administered with the compounds of the
disclosure in co-therapy. Among the useful blood thinners are, for
example: Warfarin (Coumadin) and Heparin. Blood thinners in the
various classes are described in the literature.
[0247] Anti-platelet agents can be administered with the compounds
of the disclosure in co-therapy. Among the useful anti-platelet
agents are, for example: Cyclooxygenase inhibitors (Aspirin),
Adenosine diphosphate (ADP) receptor inhibitors [Clopidogrel
(Plavix), Ticlopidine (Ticlid)], Phosphodiesterase inhibitors
[Cilostazol (Pletal)], Glycoprotein IIB/IIIA inhibitors [Abciximab
(ReoPro), Eptifibatide (Integrilin), Tirofiban (Aggrastat),
Defibrotide], Adenosine reuptake inhibitors [Dipyridamole
(Persantine)]. Anti-platelet agents in the various classes are
described in the literature.
[0248] Lipid-lowering agents can be administered with the compounds
of the disclosure in co-therapy. Among the useful lipid-lowering
agents are, for example: Statins (HMG CoA reductase inhibitors),
[Atorvastatin (Lipitor), Fluvastatin (Lescol), Lovastatin (Mevacor,
Altoprev), Pravastatin (Pravachol), Rosuvastatin Calcium (Crestor),
Simvastatin (Zocor)], Selective cholesterol absorption inhibitors
[ezetimibe (Zetia)], Resins (bile acid sequestrant or bile
acid-binding drugs) [Cholestyramine (Questran, Questran Light,
Prevalite, Locholest, Locholest Light), Colestipol (Colestid),
Colesevelam Hcl (WelChol)], Fibrates (Fibric acid derivatives)
[Gemfibrozil (Lopid), Fenofibrate (Antara, Lofibra, Tricor, and
Triglide), Clofibrate (Atromid-S)], Niacin (Nicotinic acid).
Lipid-lowering agents in the various classes are described in the
literature.
[0249] The compounds of the disclosure can be used in combination
with peptides or peptide analogs that activate the Guanylate
Cyclase-receptor in the intestine and results in elevation of the
intracellular second messenger, or cyclic guanosine monophosphate
(cGMP), with increased chloride and bicarbonate secretion into the
intestinal lumen and concomitant fluid secretion. Example of such
peptides are Linaclotide (MD-1100 Acetate), endogenous hormones
guanylin and uroguanylin and enteric bacterial peptides of the heat
stable enterotoxin family (ST peptides) and those described in U.S.
Pat. No. 5,140,102, U.S. Pat. No. 5,489,670, U.S. Pat. No.
5,969,097, WO 2006/001931A2, WO 2008/002971A2, WO 2008/106429A2, US
2008/0227685A1 and U.S. Pat. No. 7,041,786, the entire contents of
which are incorporated herein by reference for all relevant and
consistent purposes.
[0250] The compounds of the disclosure can be used in combination
with type-2 chloride channel agonists, such as Amitiza
(Lubiprostone) and other related compounds described in U.S. Pat.
No. 6,414,016, the entire contents of which are incorporated herein
by reference for all relevant and consistent purposes.
[0251] The compounds described herein can be used in combination
therapy with agents used for the treatment of obesity, T2DM,
metabolic syndrome and the like. Among the useful agents include:
insulin; insulin secretagogues, such as sulphonylureas;
glucose-lowering effectors, such as metformin; activators of the
peroxisome proliferator-activated receptor .gamma. (PPAR.gamma.),
such as the thiazolidinediones; incretin-based agents including
dipeptidyl peptidase-4 inhibitors such as sitagliptin, and
synthetic incretin mimetics such as liraglutide and exenatide;
alpha-glucosidase inhibitors, such as acarbose; glinides, such as
repaglinide and nateglinide, and the like.
[0252] The compounds of the disclosure can be used in combination
with P2Y2 receptor agonists, such as those described in EP
1196396B1 and U.S. Pat. No. 6,624,150, the entire contents of which
are incorporated herein by reference for all relevant and
consistent purposes.
[0253] Other agents include natriuretic peptides such as
nesiritide, a recombinant form of brain-natriuretic peptide (BNP)
and an atrial-natriuretic peptide (ANP). Vasopressin receptor
antagonists such as tolvaptan and conivaptan may be co-administered
as well as phosphate binders such as renagel, renleva, phoslo and
fosrenol. Other agents include phosphate transport inhibitors (as
described in U.S. Pat. Nos. 4,806,532; 6,355,823; 6,787,528;
7,119,120; 7,109,184; U.S. Pat. Pub. No. 2007/021509; 2006/0280719;
2006/0217426; International Pat. Pubs. WO 2001/005398, WO
2001/087294, WO 2001/082924, WO 2002/028353, WO 2003/048134, WO
2003/057225, WO2003/080630, WO 2004/085448, WO 2004/085382;
European Pat. Nos. 1465638 and 1485391; and JP Patent No.
2007131532, or phosphate transport antagonists such as
Nicotinamide
2. Gastrointestinal Tract Disorders
[0254] As previously noted, the compounds described herein can be
used alone or in combination with other agents. For example, the
compounds can be administered together with an analgesic peptide or
compound. The analgesic peptide or compound can be covalently
attached to a compound described herein or it can be a separate
agent that is administered together with or sequentially with a
compound described herein in a combination therapy.
[0255] Combination therapy can be achieved by administering two or
more agents, e.g., a substantially non-permeable or substantially
non-bioavailable NHE-inhibiting compound described herein and an
analgesic peptide or compound, each of which is formulated and
administered separately, or by administering two or more agents in
a single formulation. Other combinations are also encompassed by
combination therapy. For example, two agents can be formulated
together and administered in conjunction with a separate
formulation containing a third agent. While the two or more agents
in the combination therapy can be administered simultaneously, they
need not be. For example, administration of a first agent (or
combination of agents) can precede administration of a second agent
(or combination of agents) by minutes, hours, days, or weeks. Thus,
the two or more agents can be administered within minutes of each
other or within 1, 2, 3, 6, 9, 12, 15, 18, or 24 hours of each
other or within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14 days of each
other or within 2, 3, 4, 5, 6, 7, 8, 9, or weeks of each other. In
some cases even longer intervals are possible. While in many cases
it is desirable that the two or more agents used in a combination
therapy be present in within the patient's body at the same time,
this need not be so.
[0256] Combination therapy can also include two or more
administrations of one or more of the agents used in the
combination. For example, if agent X and agent Y are used in a
combination, one could administer them sequentially in any
combination one or more times, e.g., in the order X--Y--X, X--X--Y,
Y--X--Y, Y--Y--X, X--X--Y--Y, etc.
[0257] The compounds described herein can be used in combination
therapy with an analgesic agent, e.g., an analgesic compound or an
analgesic peptide. The analgesic agent can optionally be covalently
attached to a compound described herein. Among the useful analgesic
agents are, for example: Ca channel blockers, 5HT3 agonists (e.g.,
MCK-733), 5HT4 agonists (e.g., tegaserod, prucalopride), and 5HT1
receptor antagonists, opioid receptor agonists (loperamide,
fedotozine, and fentanyl), NK1 receptor antagonists, CCK receptor
agonists (e.g., loxiglumide), NK1 receptor antagonists, NK3
receptor antagonists, norepinephrine-serotonin reuptake inhibitors
(NSR1), vanilloid and cannabanoid receptor agonists, and
sialorphin. Analgesics agents in the various classes are described
in the literature.
[0258] Opioid receptor antagonists and agonists can be administered
with the compounds of the disclosure in co-therapy or linked to the
compound of the disclosure, e.g., by a covalent bond. For example,
opioid receptor antagonists such as naloxone, naltrexone, methyl
nalozone, nalmefene, cypridime, beta funaltrexamine, naloxonazine,
naltrindole, and nor-binaltorphimine are thought to be useful in
the treatment of opioid-induced constipaption (OIC). It can be
useful to formulate opioid antagonists of this type in a delayed or
sustained release formulation, such that initial release of the
antagonist is in the mid to distal small intestine and/or ascending
colon. Such antagonists are described in U.S. Pat. No. 6,734,188
(WO 01/32180 A2), the entire contents of which are incorporated
herein by reference for all relevant and consistent purposes.
Enkephalin pentapeptide (HOE825; Tyr-D-Lys-Gly-Phe-L-homoserine) is
an agonist of the .mu.- and .gamma.-opioid receptors and is thought
to be useful for increasing intestinal motility (Eur. J. Pharm.,
219:445, 1992), and this peptide can be used in conjunction with
the compounds of the disclosure. Also useful is trimebutine which
is thought to bind to mu/delta/kappa opioid receptors and activate
release of motilin and modulate the release of gastrin, vasoactive
intestinal peptide, gastrin and glucagons. K-opioid receptor
agonists such as fedotozine, ketocyclazocine, and compounds
described in US 2005/0176746 (WO 03/097051 A2), the entire contents
of which are incorporated herein by reference for all relevant and
consistent purposes, can be used with or linked to the compounds of
the disclosure. In addition, .mu.-opioid receptor agonists, such as
morphine, diphenyloxylate, frakefamide
(H-Tyr-D-Ala-Phe(F)-Phe-NH.sub.2; disclosed in WO 01/019849 A1, the
entire contents of which are incorporated herein by reference for
all relevant and consistent purposes) and loperamide can be
used.
[0259] Tyr-Arg (kyotorphin) is a dipeptide that acts by stimulating
the release of met-enkephalins to elicit an analgesic effect (J.
Biol. Chem. 262:8165, 1987). Kyotorphin can be used with or linked
to the compounds of the disclosure. CCK receptor agonists such as
caerulein from amphibians and other species are useful analgesic
agents that can be used with or linked to the compounds of the
disclosure.
[0260] Conotoxin peptides represent a large class of analgesic
peptides that act at voltage gated Ca channels, NMDA receptors or
nicotinic receptors. These peptides can be used with or linked to
the compounds of the disclosure.
[0261] Peptide analogs of thymulin (U.S. Pat. No. 7,309,690 or FR
2830451, the entire contents of which are incorporated herein by
reference for all relevant and consistent purposes) can have
analgesic activity and can be used with or linked to the compounds
of the disclosure.
[0262] CCK (CCKa or CCKb) receptor antagonists, including
loxiglumide and dexloxiglumide (the R-isomer of loxiglumide) (U.S.
Pat. No. 5,130,474 or WO 88/05774, the entire contents of which are
incorporated herein by reference for all relevant and consistent
purposes) can have analgesic activity and can be used with or
linked to the compounds of the disclosure.
[0263] Other useful analgesic agents include 5-HT4 agonists such as
tegaserod/zelnorm and lirexapride. Such agonists are described in:
EP1321142 A1, WO 03/053432A1, EP 505322 A1, EP 505322 B1, EP 507672
A1, EP 507672 B1, U.S. Pat. No. 5,510,353 and U.S. Pat. No.
5,273,983, the entire contents of which are incorporated herein by
reference for all relevant and consistent purposes.
[0264] Calcium channel blockers such as ziconotide and related
compounds described in, for example, EP 625162B1, U.S. Pat. No.
5,364,842, U.S. Pat. No. 5,587,454, U.S. Pat. No. 5,824,645, U.S.
Pat. No. 5,859,186, U.S. Pat. No. 5,994,305, U.S. Pat. No.
6,087,091, U.S. Pat. No. 6,136,786, WO 93/13128 A1, EP 1336409 A1,
EP 835126 A1, EP 835126 B1, U.S. Pat. No. 5,795,864, U.S. Pat. No.
5,891,849, U.S. Pat. No. 6,054,429, WO 97/01351 A1, the entire
contents of which are incorporated herein by reference for all
relevant and consistent purposes, can be used with or linked to the
compounds of the disclosure.
[0265] Various antagonists of the NK-1, NK-2, and NK-3 receptors
(for a review see Giardina et al. 2003 Drugs 6:758) can be can be
used with or linked to the compounds of the disclosure.
[0266] NK1 receptor antagonists such as: aprepitant (Merck & Co
Inc), vofopitant, ezlopitant (Pfizer, Inc.), R-673 (Hoffmann-La
Roche Ltd), SR-14033 and related compounds described in, for
example, EP 873753 A1, U.S. 20010006972 A1, U.S. 20030109417 A1, WO
01/52844 A1, the entire contents of which are incorporated herein
by reference for all relevant and consistent purposes, can be used
with or linked to the compounds of the disclosure.
[0267] NK-2 receptor antagonists such as nepadutant (Menarini
Ricerche SpA), saredutant (Sanofi-Synthelabo), SR-144190
(Sanofi-Synthelabo) and UK-290795 (Pfizer Inc) can be used with or
linked to the compounds of the disclosure.
[0268] NK3 receptor antagonists such as osanetant
(Sanofi-Synthelabo), talnetant and related compounds described in,
for example, WO 02/094187 A2, EP 876347 A1, WO 97/21680 A1, U.S.
Pat. No. 6,277,862, WO 98/11090, WO 95/28418, WO 97/19927, and
Boden et al. (J Med. Chem. 39:1664-75, 1996), the entire contents
of which are incorporated herein by reference for all relevant and
consistent purposes, can be used with or linked to the compounds of
the disclosure.
[0269] Norepinephrine-serotonin reuptake inhibitors such as
milnacipran and related compounds described in WO 03/077897 A1, the
entire contents of which are incorporated herein by reference for
all relevant and consistent purposes, can be used with or linked to
the compounds of the disclosure.
[0270] Vanilloid receptor antagonists such as arvanil and related
compounds described in WO 01/64212 A1, the entire contents of which
are incorporated herein by reference for all relevant and
consistent purposes, can be used with or linked to the compounds of
the disclosure.
[0271] The compounds can be used in combination therapy with a
phosphodiesterase inhibitor (examples of such inhibitors can be
found in U.S. Pat. No. 6,333,354, the entire contents of which are
incorporated herein by reference for all relevant and consistent
purposes).
[0272] The compounds can be used alone or in combination therapy to
treat disorders associated with chloride or bicarbonate secretion
that may lead to constipation, e.g., Cystic Fibrosis.
[0273] The compounds can also or alternatively be used alone or in
combination therapy to treat calcium-induced constipation effects.
Constipation is commonly found in the geriatric population,
particularly patients with osteoporosis who have to take calcium
supplements. Calcium supplements have shown to be beneficial in
ostoporotic patients to restore bone density but compliance is poor
because of constipation effects associated therewith.
[0274] The compounds of the current disclosure have can be used in
combination with an opioid. Opioid use is mainly directed to pain
relief, with a notable side-effect being GI disorder, e.g.
constipation. These agents work by binding to opioid receptors,
which are found principally in the central nervous system and the
gastrointestinal tract. The receptors in these two organ systems
mediate both the beneficial effects, and the undesirable side
effects (e.g. decrease of gut motility and ensuing constipation).
Opioids suitable for use typically belong to one of the following
exemplary classes: natural opiates, alkaloids contained in the
resin of the opium poppy including morphine, codeine and thebaine;
semi-synthetic opiates, created from the natural opioids, such as
hydromorphone, hydrocodone, oxycodone, oxymorphone, desomorphine,
diacetylmorphine (Heroin), nicomorphine, dipropanoylmorphine,
benzylmorphine and ethylmorphine; fully synthetic opioids, such as
fentanyl, pethidine, methadone, tramadol and propoxyphene;
endogenous opioid peptides, produced naturally in the body, such as
endorphins, enkephalins, dynorphins, and endomorphins.
[0275] The compound of the disclosure can be used alone or in
combination therapy to alleviate GI disorders encountered with
patients with renal failure (stage 3-5). Constipation is the second
most reported symptom in that category of patients (Murtagh et al.,
2006; Murtagh et al., 2007a; Murtagh et al., 2007b). Without being
held by theory, it is believed that kidney failure is accompanied
by a stimulation of intestinal Na re-absorption (Hatch and Freel,
2008). A total or partial inhibition of such transport by
administration of the compounds of the disclosure can have a
therapeutic benefit to improve GI transit and relieve abdominal
pain. In that context, the compounds of the disclosure can be used
in combination with Angiotensin-modulating agents: Angiotensin
Converting Enzyme (ACE) inhibitors (e.g. captopril, enalopril,
lisinopril, ramipril) and Angiotensin II receptor antagonist
therapy (also referred to as AT.sub.1-antagonists or angiotensin
receptor blockers, or ARB's); diuretics such as loop diuretics
(e.g. furosemide, bumetanide), Thiazide diuretics (e.g.
hydrochlorothiazide, chlorthalidone, chlorthiazide) and
potassium-sparing diuretics: amiloride; beta blockers: bisoprolol,
carvedilol, nebivolol and extended-release metoprolol; positive
inotropes: Digoxin, dobutamine; phosphodiesterase inhibitors such
as milrinone; alternative vasodilators: combination of isosorbide
dinitrate/hydralazine; aldosterone receptor antagonists:
spironolactone, eplerenone; natriuretic peptides: Nesiritide, a
recombinant form of brain-natriuretic peptide (BNP),
atrial-natriuretic peptide (ANP); vasopressin receptor antagonists:
Tolvaptan and conivaptan; phosphate binder (Renagel, Renleva,
Phoslo, Fosrenol); phosphate transport inhibitor such as those
described in U.S. Pat. No. 4,806,532, U.S. Pat. No. 6,355,823, U.S.
Pat. No. 6,787,528, WO 2001/005398, WO 2001/087294, WO 2001/082924,
WO 2002/028353, WO 2003/048134, WO 2003/057225, U.S. Pat. No.
7,119,120, EP 1465638, US Appl. 2007/021509, WO 2003/080630, U.S.
Pat. No. 7,109,184, US Appl. 2006/0280719, EP 1485391, WO
2004/085448, WO 2004/085382, US Appl. 2006/0217426, JP 2007/131532,
the entire contents of which are incorporated herein by reference
for all relevant and consistent purposes, or phosphate transport
antagonist (Nicotinamide).
[0276] The compounds of the disclosure can be used in combination
with peptides or peptide analogs that activate the Guanylate
Cyclase-receptor in the intestine and results in elevation of the
intracellular second messenger, or cyclic guanosine monophosphate
(cGMP), with increased chloride and bicarbonate secretion into the
intestinal lumen and concomitant fluid secretion. Example of such
peptides are Linaclotide (MD-1100 Acetate), endogenous hormones
guanylin and uroguanylin and enteric bacterial peptides of the heat
stable enterotoxin family (ST peptides) and those described in U.S.
Pat. No. 5,140,102, U.S. Pat. No. 5,489,670, U.S. Pat. No.
5,969,097, WO 2006/001931A2, WO 2008/002971A2, WO 2008/106429A2, US
2008/0227685A1 and U.S. Pat. No. 7,041,786, the entire contents of
which are incorporated herein by reference for all relevant and
consistent purposes.
[0277] The compounds of the disclosure can be used in combination
with type-2 chloride channel agonists, such as Amitiza
(Lubiprostone) and other related compounds described in U.S. Pat.
No. 6,414,016, the entire contents of which are incorporated herein
by reference for all relevant and consistent purposes.
[0278] The compounds of the disclosure can be used in combination
with P2Y2 receptor agonists, such as those described in EP
1196396B1 and U.S. Pat. No. 6,624,150, the entire contents of which
are incorporated herein by reference for all relevant and
consistent purposes.
[0279] The compounds of the disclosure can be used in combination
with laxative agents such as bulk-producing agents, e.g. psyllium
husk (Metamucil), methylcellulose (Citrucel), polycarbophil,
dietary fiber, apples, stool softeners/surfactant such as docusate
(Colace, Diocto); hydrating agents (osmotics), such as dibasic
sodium phosphate, magnesium citrate, magnesium hydroxide (Milk of
magnesia), magnesium sulfate (which is Epsom salt), monobasic
sodium phosphate, sodium biphosphate; hyperosmotic agents: glycerin
suppositories, sorbitol, lactulose, and polyethylene glycol (PEG).
The compounds of the disclosure can be also be used in combination
with agents that stimulate gut peristalsis, such as Bisacodyl
tablets (Dulcolax), Casanthranol, Senna and Aloin, from Aloe
Vera.
[0280] In one embodiment, the compounds of the disclosure
accelerate gastrointestinal transit, and more specifically in the
colon, without substantially affecting the residence time in the
stomach, i.e. with no significant effect on the gastric emptying
time. Even more specifically the compounds of the invention restore
colonic transit without the side-effects associated with delayed
gastric emptying time, such as nausea. The GI and colonic transit
are measured in patients using methods reported in, for example:
Burton D D, Camilleri M, Mullan B P, et al., J. Nucl. Med., 1997;
38:1807-1810; Cremonini F, Mullan B P, Camilleri M, et al.,
Aliment. Pharmacol. Ther., 2002; 16:1781-1790; Camilleri M,
Zinsmeister A R, Gastroenterology, 1992; 103:36-42; Bouras E P,
Camilleri M, Burton D D, et al., Gastroenterology, 2001;
120:354-360; Coulie B, Szarka L A, Camilleri M, et al.,
Gastroenterology, 2000; 119:41-50; Prather C M, Camilleri M,
Zinsmeister A R, et al., Gastroenterology, 2000; 118:463-468; and,
Camilleri M, McKinzie S, Fox J, et al., Clin. Gastroenterol.
Hepatol., 2004; 2:895-904.
C. Polymer Combination Therapy
[0281] The NHE-inhibiting compounds described therein may be
administered to patients in need thereof in combination with a
fluid-absorbing polymer ("FAP"). The intestinal fluid-absorbing
polymers useful for administration in accordance with embodiments
of the present disclosure may be administered orally in combination
with non-absorbable NHE-inhibiting compounds (e.g., a NHE-3
inhibitor) to absorb the intestinal fluid resulting from the action
of the sodium transport inhibitors. Such polymers swell in the
colon and bind fluid to impart a consistency to stools that is
acceptable for patients. The fluid-absorbing polymers described
herein may be selected from polymers with laxative properties, also
referred to as bulking agents (i.e., polymers that retain some of
the intestinal fluid in the stools and impart a higher degree of
hydration in the stools and facilitate transit). The
fluid-absorbing polymers may also be optionally selected from
pharmaceutical polymers with anti-diarrhea function, i.e., agents
that maintain some consistency to the stools to avoid watery stools
and potential incontinence.
[0282] The ability of the polymer to maintain a certain consistency
in stools with a high content of fluid can be characterized by its
"water holding power." Wenzl et al. (in Determinants of decreased
fecal consistency in patients with diarrhea; Gastroenterology, v.
108, no. 6, p. 1729-1738 (1995)) studied the determinants that
control the consistency of stools of patients with diarrhea and
found that they were narrowly correlated with the water holding
power of the feces. The water holding power is determined as the
water content of given stools to achieve a certain level of
consistency (corresponding to "formed stool" consistency) after the
reconstituted fecal matter has been centrifuged at a certain g
number. Without being held to any particular theory, has been found
that the water holding power of the feces is increased by ingestion
of certain polymers with a given fluid absorbing profile. More
specifically, it has been found that the water-holding power of
said polymers is correlated with their fluid absorbancy under load
(AUL); even more specifically the AUL of said polymers is greater
than 15 g of isotonic fluid/g of polymer under a static pressure of
5 kPa, even more preferably under a static pressure of 10 kPa.
[0283] The FAP utilized in the treatment method of the present
disclosure preferably has a AUL of at least about 10 g, about 15 g,
about 20 g, about 25 g or more of isotonic fluid/g of polymer under
a static pressure of about 5 kPa, and preferably about 10 kPA, and
may have a fluid absorbency of about 20 g, about 25 g or more, as
determined using means generally known in the art. Additionally or
alternatively, the FAP may impart a minimum consistency to fecal
matter and, in some embodiments, a consistency graded as "soft" in
the scale described in the test method below, when fecal non
water-soluble solid fraction is from 10% to 20%, and the polymer
concentration is from 1% to 5% of the weight of stool. The
determination of the fecal non water-soluble solid fraction of
stools is described in Wenz et al. The polymer may be uncharged or
may have a low charge density (e.g., 1-2 meq/gr). Alternatively or
in addition, the polymer may be delivered directly to the colon
using known delivery methods to avoid premature swelling in the
esophagus.
[0284] In one embodiment of the present disclosure, the FAP is a
"superabsorbent" polymer (i.e., a lightly crosslinked, partially
neutralized polyelectrolyte hydrogel similar to those used in baby
diapers, feminine hygiene products, agriculture additives, etc.).
Superabsorbent polymers may be made of a lightly crosslinked
polyacrylate hydrogel. The swelling of the polymer is driven
essentially by two effects: (i) the hydration of the polymer
backbone and entropy of mixing and (ii) the osmotic pressure
arising from the counter-ions (e.g., Na ions) within the gel. The
gel swelling ratio at equilibrium is controlled by the elastic
resistance inherent to the polymer network and by the chemical
potential of the bathing fluid, i.e., the gel will de-swell at
higher salt concentration because the background electrolyte will
reduce the apparent charge density on the polymer and will reduce
the difference of free ion concentrations inside and outside the
gel that drives osmotic pressure. The swelling ratio SR (g of fluid
per g of dry polymer and synonymously "fluid absorbency") may vary
from 1000 in pure water down to 30 in 0.9% NaCl solution
representative of physiological saline (i.e., isotonic). SR may
increase with the degree of neutralization and may decrease with
the crosslinking density. SR generally decreases with an applied
load with the extent of reduction dependent on the strength of the
gel, i.e., the crosslinking density. The salt concentration within
the gel, as compared with the external solution, may be lower as a
result of the Donnan effect due to the internal electrical
potential.
[0285] The fluid-absorbing polymer may include crosslinked
polyacrylates which are fluid absorbent such as those prepared from
.alpha.,.beta.-ethylenically unsaturated monomers, such as
monocarboxylic acids, polycarboxylic acids, acrylamide and their
derivatives. These polymers may have repeating units of acrylic
acid, methacrylic acid, metal salts of acrylic acid, acrylamide,
and acrylamide derivatives (such as
2-acrylamido-2-methylpropanesulfonic acid) along with various
combinations of such repeating units as copolymers. Such
derivatives include acrylic polymers which include hydrophilic
grafts of polymers such as polyvinyl alcohol. Examples of suitable
polymers and processes, including gel polymerization processes, for
preparing such polymers are disclosed in U.S. Pat. Nos. 3,997,484;
3,926,891; 3,935,099; 4,090,013; 4,093,776; 4,340,706; 4,446,261;
4,683,274; 4,459,396; 4,708,997; 4,076,663; 4,190,562; 4,286,082;
4,857,610; 4,985,518; 5,145,906; 5,629,377 and 6,908,609 which are
incorporated herein by reference for all relevant and consistent
purposes (in addition, see Buchholz, F. L. and Graham, A. T.,
"Modern Superabsorbent Polymer Technology," John Wiley & Sons
(1998), which is also incorporated herein by reference for all
relevant and consistent purposes). A class of preferred polymers
for treatment in combination with NHE-inhibitors is
polyelectrolytes.
[0286] The degree of crosslinking can vary greatly depending upon
the specific polymer material; however, in most applications the
subject superabsorbent polymers are only lightly crosslinked, that
is, the degree of crosslinking is such that the polymer can still
absorb over 10 times its weight in physiological saline (i.e., 0.9%
saline). For example, such polymers typically include less than
about 0.2 mole % crosslinking agent.
[0287] In some embodiments, the FAP's utilized for treatment are
Calcium Carbophil (Registry Number: 9003-97-8, also referred as
Carbopol EX-83), and Carpopol 934P.
[0288] In some embodiments, the fluid-absorbing polymer is prepared
by high internal phase emulsion ("HIPE") processes. The HIPE
process leads to polymeric foam slabs with a very large porous
fraction of interconnected large voids (about 100 microns) (i.e.,
open-cell structures). This technique produces flexible and
collapsible foam materials with exceptional suction pressure and
fluid absorbency (see U.S. Pat. Nos. 5,650,222; 5,763,499 and
6,107,356, which are incorporated herein for all relevant and
consistent purposes). The polymer is hydrophobic and, therefore,
the surface should be modified so as to be wetted by the aqueous
fluid. This is accomplished by post-treating the foam material by a
surfactant in order to reduce the interfacial tension. These
materials are claimed to be less compliant to loads, i.e., less
prone to de-swelling under static pressure.
[0289] In some embodiments, fluid-absorbing gels are prepared by
aqueous free radical polymerization of acrylamide or a derivative
thereof, a crosslinker (e.g., methylene-bis-acrylamide) and a free
radical initiator redox system in water. The material is obtained
as a slab. Typically the swelling ratio of crosslinked
polyacrylamide at low crosslinking density (e.g., 2%-4% expressed
as weight % of methylene-bis-acrylamide) is between 25 and 40 (F.
Horkay, Macromolecules, 22, pp. 2007-09 (1989)). The swelling
properties of these polymers have been extensively studied and are
essentially the same of those of crosslinked polyacrylic acids at
high salt concentration. Under those conditions, the osmotic
pressure is null due to the presence of counter-ions and the
swelling is controlled by the free energy of mixing and the network
elastic energy. Stated differently, a crosslinked polyacrylamide
gel of same crosslink density as a neutralized polyacrylic acid
will exhibit the same swelling ratio (i.e., fluid absorbing
properties) and it is believed the same degree of deswelling under
pressure, as the crosslinked polyelectrolyte at high salt content
(e.g., 1 M). The properties (e.g., swelling) of neutral hydrogels
will not be sensitive to the salt environment as long as the
polymer remains in good solvent conditions. Without being held to
any particular theory, it is believed that the fluid contained
within the gel has the same salt composition than the surrounding
fluid (i.e., there is no salt partitioning due to Donnan
effect).
[0290] Another subclass of fluid-absorbing polymers that may be
utilized is hydrogel materials that include N-alkyl acrylamide
polymers (e.g., N-isopropylacrylamide (NIPAM)). The corresponding
aqueous polyNIPAM hydrogel shows a temperature transition at about
35.degree. C. Above this temperature the hydrogel may collapse. The
mechanism is generally reversible and the gel re-swells to its
original swelling ratio when the temperature reverts to room
temperature. This allows production of nanoparticles by emulsion
polymerization (R. Pelton, Advances in Colloid and Interface
Science, 85, pp. 1-33, (2000)). The swelling characteristics of
poly-NIPAM nanoparticles below the transition temperature have been
reported and are similar to those reported for bulk gel of
polyNIPAM and equivalent to those found for polyacrylamide (i.e.
30-50 g/g) (W. McPhee, Journal of Colloid and Interface Science,
156, pp. 24-30 (1993); and, K. Oh, Journal of Applied Polymer
Science, 69, pp. 109-114 (1997)).
[0291] In some embodiments, the FAP utilized for treatment in
combination with a NHE-inhibitor is a superporous gel that may
delay the emptying of the stomach for the treatment of obesity (J.
Chen, Journal of Controlled Release, 65, pp. 73-82 (2000), or to
deliver proteins. Polyacrylate-based SAP's with a macroporous
structure may also be used. Macroporous SAP and superporous gels
differ in that the porous structure remains almost intact in the
dry state for superporous gels, but disappears upon drying for
macroporous SAP's. The method of preparation is different although
both methods use a foaming agent (e.g., carbonate salt that
generates CO.sub.2 bubbles during polymerization). Typical swelling
ratios, SR, of superporous materials are around 10. Superporous
gels keep a large internal pore volume in the dry state.
[0292] Macroporous hydrogels may also be formed using a method
whereby polymer phase separation in induced by a non-solvent. The
polymer may be poly-NIPAM and the non-solvent utilized may be
glucose (see, e.g., Z. Zhang, J. Org. Chem., 69, 23 (2004)) or NaCl
(see, e.g., Cheng et al., Journal of Biomedical Materials
Research--Part A, Vol. 67, Issue 1, 1 Oct. 2003, Pages 96-103). The
phase separation induced by the presence of NaCl leads to an
increase in swelling ratio. These materials are preferred if the
swelling ratio of the material, SR, is maintained in salt isotonic
solution and if the gels do not collapse under load. The
temperature of "service" should be shifted beyond body temperature,
e.g. by diluting NIPAM in the polymer with monomer devoid of
transition temperature phenomenon.
[0293] In some embodiments, the fluid-absorbing polymer may be
selected from certain naturally-occurring polymers such as those
containing carbohydrate moieties. In a preferred embodiment, such
carbohydrate-containing hydrogels are non-digestible, have a low
fraction of soluble material and a high fraction of gel-forming
materials. In some embodiments, the fluid-absorbing polymer is
selected from xanthan, guar, wellan, hemicelluloses,
alkyl-cellulose, hydro-alkyl-cellulose, carboxy-alkyl-cellulose,
carrageenan, dextran, hyaluronic acid and agarose. In a preferred
embodiment, the gel forming polymer is psyllium. Psyllium (or
"ispaghula") is the common name used for several members of the
plant genus Plantago whose seeds are used commercially for the
production of mucilage. Most preferably, the fluid-absorbing
polymer is in the gel-forming fraction of psyllium, i.e., a neutral
saccharide copolymer of arabinose (25%) and xylose (75%) as
characterized in (J. Marlett, Proceedings of the Nutrition Society,
62, pp. 2-7-209 (2003); and, M. Fischer, Carbohydrate Research,
339, 2009-2012 (2004)), and further described in U.S. Pat. Nos.
6,287,609; 7,026,303; 5,126,150; 5,445,831; 7,014,862; 4,766,004;
4,999,200, each of which is incorporated herein for all relevant
and consistent purposes, and over-the-counter psillium-containing
agents such as those marketed under the name Metamucil (The Procter
and Gamble company). Preferably the a psyllium-containing dosage
form is suitable for chewing, where the chewing action
disintegrates the tablet into smaller, discrete particles prior to
swallowing but which undergoes minimal gelling in the mouth, and
has acceptable mouthfeel and good aesthetics as perceived by the
patient.
[0294] The psyllium-containing dosage form includes physically
discrete unit suitable as a unitary dosage for human subjects and
other mammals, each containing a predetermined quantity of active
material (e.g. the gel-forming polysaccharide) calculated to
produce the desired therapeutic effect. Solid oral dosage forms
that are suitable for the present compositions include tablets,
pills, capsules, lozenges, chewable tablets, troches, cachets,
pellets, wafer and the like.
[0295] In some embodiments, the FAP is a polysaccharide particle
wherein the polysaccharide component includes xylose and arabinose.
The ratio of the xylose to the arabinose may be at least about 3:1
by weight, as described in U.S. Pat. Nos. 6,287,609; 7,026,303 and
7,014,862, each of which is incorporated herein for all relevant
and consistent purposes.
[0296] The fluid-absorbing polymers described herein may be used in
combination with the NHE-inhibiting compound or a pharmaceutical
composition containing it. The NHE-inhibiting compound and the FAP
may also be administered with other agents including those
described under the heading "Combination Therapies" without
departing from the scope of the present disclosure. As described
above, the NHE-inhibiting compound may be administered alone
without use of a fluid-absorbing polymer to resolve symptoms
without eliciting significant diarrhea or fecal fluid secretion
that would require the co-administration of a fluid-absorbing
polymer.
[0297] The fluid-absorbing polymers described herein may be
selected so as to not induce any substantial interaction with the
NHE-inhibiting compound or a pharmaceutical composition containing
it. As used herein, "no substantial interaction" generally means
that the co-administration of the FAP polymer would not
substantially alter (i.e., neither substantially decrease nor
substantially increase) the pharmacological property of the
NHE-inhibiting compounds administered alone. For example, FAPs
containing negatively charged functionality, such as carboxylates,
sulfonates, and the like, may potentially interact ionically with
positively charged NHE-inhibiting compounds, preventing the
inhibitor from reaching its pharmacological target. In addition, it
may be possible that the shape and arrangement of functionality in
a FAP could act as a molecular recognition element, and sequestor
NHE-inhibiting compounds via "host-guest" interactions via the
recognition of specific hydrogen bonds and/or hydrophobic regions
of a given inhibitor. Accordingly, in various embodiments of the
present disclosure, the FAP polymer may be selected, for
co-administration or use with a compound of the present disclosure,
to ensure that (i) it does not ionically interact with or bind with
the compound of the present disclosure (by means of, for example, a
moiety present therein possessing a charge opposite that of a
moiety in the compound itself), and/or (ii) it does not possess a
charge and/or structural conformation (or shape or arrangement)
that enables it to establish a "host-guest" interaction with the
compound of the present disclosure (by means of, for example, a
moiety present therein that may act as a molecular recognition
element and sequester the NHE inhibitor or inhibiting moiety of the
compound).
D. Dosage
[0298] It is to be noted that, as used herein, an "effective
amount" (or "pharmaceutically effective amount") of a compound
disclosed herein, is a quantity that results in a beneficial
clinical outcome of the condition being treated with the compound
compared with the absence of treatment. The amount of the compound
or compounds administered will depend on the degree, severity, and
type of the disease or condition, the amount of therapy desired,
and the release characteristics of the pharmaceutical formulation.
It will also depend on the subject's health, size, weight, age, sex
and tolerance to drugs. Typically, the compound is administered for
a sufficient period of time to achieve the desired therapeutic
effect.
[0299] In embodiments wherein both an NHE-inhibitor compound and a
fluid-absorbing polymer are used in the treatment protocol, the
NHE-inhibiting compound and FAP may be administered together or in
a "dual-regimen" wherein the two therapeutics are dosed and
administered separately. When the NHE-inhibiting compound and the
fluid-absorbing polymer are dosed separately, the typical dosage
administered to the subject in need of the NHE-inhibiting compound
is typically from about 5 mg per day and about 5000 mg per day and,
in other embodiments, from about 50 mg per day and about 1000 mg
per day. Such dosages may induce fecal excretion of sodium (and its
accompanying anions), from about 10 mmol up to about 250 mmol per
day, from about 20 mmol to about 70 mmol per day or even from about
30 mmol to about 60 mmol per day.
[0300] The typical dose of the fluid-absorbing polymer is a
function of the extent of fecal secretion induced by the
non-absorbable NHE-inhibiting compound. Typically the dose is
adjusted according to the frequency of bowel movements and
consistency of the stools. More specifically the dose is adjusted
so as to avoid liquid stools and maintain stool consistency as
"soft" or semi-formed, or formed. To achieve the desired stool
consistency and provide abdominal relief to patients, typical
dosage ranges of the fluid-absorbing polymer to be administered in
combination with the NHE-inhibiting compound, are from about 2 g to
about 50 g per day, from about 5 g to about 25 g per day or even
from about 10 g to about 20 g per day. When the NHE-inhibiting
compound and the FAP are administered as a single dosage regimen,
the daily uptake may be from about 2 g to about 50 g per day, from
about 5 g to about 25 g per day, or from about 10 g to about 20 g
per day, with a weight ratio of NHE-inhibiting compound to
fluid-absorbing polymer being from about 1:1000 to 1:10 or even
from about 1:500 to 1:5 or about 1:100 to 1:5.
[0301] A typical dosage of the substantially impermeable or
substantially systemically non-bioavailable, NHE-inhibiting
compound when used alone without a FAP may be between about 0.2 mg
per day and about 2 g per day, or between about 1 mg and about 1 g
per day, or between about 5 mg and about 500 mg, or between about
10 mg and about 250 mg per day, which is administered to a subject
in need of treatment.
[0302] The frequency of administration of therapeutics described
herein may vary from once-a-day (QD) to twice-a-day (BID) or
thrice-a-day (TID), etc., the precise frequency of administration
varying with, for example, the patient's condition, the dosage,
etc. For example, in the case of a dual-regimen, the NHE-inhibiting
compound could be taken once-a-day while the fluid-absorbing
polymer could be taken at each meal (TID). Furthermore, as
disclosed in U.S. Application No. 61/584,753 filed Jan. 9, 2012,
the NHE-inhibiting compound is administered twice-a-day (BID), or
thrice-a-day (TID), and in a more specific embodiment, the
NHE-inhibiting compound is administered in an amount ranging from
2-200 mg per dose BID, or 2-100 mg per dose TID. In more specific
embodiments, the NHE-inhibiting compound is administered in an
amount of about 15 mg per dose, about 30 mg per dose, or about 45
mg per dose, and in a more specific embodiment, in an amount of 15
mg per dose, 30 mg per dose, or 45 mg per dose.
E. Modes of Administration
[0303] The substantially impermeable or substantially systemically
non-bioavailable NHE-inhibiting compounds of the present disclosure
with or without the fluid-absorbing polymers described herein may
be administered by any suitable route. The compound is preferably
administrated orally (e.g., dietary) in capsules, suspensions,
tablets, pills, dragees, liquids, gels, syrups, slurries, and the
like. Methods for encapsulating compositions (such as in a coating
of hard gelatin or cyclodextran) are known in the art (Baker, et
al., "Controlled Release of Biological Active Agents", John Wiley
and Sons, 1986). The compounds can be administered to the subject
in conjunction with an acceptable pharmaceutical carrier as part of
a pharmaceutical composition. The formulation of the pharmaceutical
composition will vary according to the route of administration
selected. Suitable pharmaceutical carriers may contain inert
ingredients which do not interact with the compound. The carriers
are biocompatible, i.e., non-toxic, non-inflammatory,
non-immunogenic and devoid of other undesired reactions at the
administration site. Examples of pharmaceutically acceptable
carriers include, for example, saline, commercially available inert
gels, or liquids supplemented with albumin, methyl cellulose or a
collagen matrix. Standard pharmaceutical formulation techniques can
be employed, such as those described in Remington's Pharmaceutical
Sciences, Mack Publishing Company, Easton, Pa.
[0304] Pharmaceutical preparations for oral use can be obtained by
combining a compound of the present disclosure with a solid
excipient, optionally grinding a resulting mixture, and processing
the mixture of granules, after adding suitable auxiliaries, if
desired, to obtain tablets or dragee cores. Suitable excipients
are, in particular, fillers such as sugars, including lactose,
sucrose, mannitol, or sorbitol; cellulose preparations such as, for
example, maize starch, wheat starch, rice starch, potato starch,
gelatin, gum tragacanth, methyl cellulose,
hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and/or
polyvinylpyrrolidone (PVP). If desired, disintegrating agents can
be added, such as cross-linked polyvinyl pyrrolidone, agar, or
alginic acid or a salt thereof such as sodium alginate.
[0305] Dragee cores are provided with suitable coatings. For this
purpose, concentrated sugar solutions can be used, which can
optionally contain gum arabic, talc, polyvinyl pyrrolidone,
carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer
solutions, and suitable organic solvents or solvent mixtures.
Dyestuffs or pigments can be added to the tablets or dragee
coatings for identification or to characterize different
combinations of active compound doses.
[0306] Pharmaceutical preparations which can be used orally include
push-fit capsules made of a suitable material, such as gelatin, as
well as soft, sealed capsules made of a suitable material, for
example, gelatin, and a plasticizer, such as glycerol or sorbitol.
The push-fit capsules can contain the active ingredients in
admixture with filler such as lactose, binders such as starches,
and/or lubricants such as talc or magnesium stearate and,
optionally, stabilizers. In soft capsules, the active compounds can
be dissolved or suspended in suitable liquids, such as fatty oils,
liquid paraffin, or liquid polyethylene glycols. In addition,
stabilizers can be added. All formulations for oral administration
should be in dosages suitable for such administration.
[0307] It will be understood that, certain compounds of the
disclosure may be obtained as different stereoisomers (e.g.,
diastereomers and enantiomers) or as isotopes and that the
disclosure includes all isomeric forms, racemic mixtures and
isotopes of the disclosed compounds and a method of treating a
subject with both pure isomers and mixtures thereof, including
racemic mixtures, as well as isotopes. Stereoisomers can be
separated and isolated using any suitable method, such as
chromatography.
F. Delayed Release
[0308] NHE proteins show considerable diversity in their patterns
of tissue expression, membrane localization and functional roles.
(See, e.g., The sodium-hydrogen exchanger-From molecule To Its Role
In Disease, Karmazyn, M., Avkiran, M., and Fliegel, L., eds.,
Kluwer Academics (2003).)
[0309] In mammals, nine distinct NHE genes (NHE-1 through -9) have
been described. Of these nine, five (NHE-1 through -5) are
principally active at the plasma membrane, whereas NHE-6, -7 and -9
reside predominantly within intracellular compartments.
[0310] NHE-1 is ubiquitously expressed and is chiefly responsible
for restoration of steady state intracellular pH following
cytosolic acidification and for maintenance of cell volume. Recent
findings show that NHE-1 is crucial for organ function and survival
(e.g., NHE-1-null mice exhibit locomotor abnormalities,
epileptic-like seizures and considerable mortality before
weaning).
[0311] In contrast with NHE-1 expressed at the basolateral side of
the nephrons and gut epithelial cells, NHE-2 through -4 are
predominantly expressed on the apical side of epithelia of the
kidney and the gastrointestinal tract. Several lines of evidence
show that NHE-3 is the major contributor of renal bulk Na+ and
fluid re-absorption by the proximal tubule. The associated
secretion of H+ by NHE-3 into the lumen of renal tubules is also
essential for about 2/3 of renal HCO3.sup.- re-absorption. Complete
disruption of NHE-3 function in mice causes a sharp reduction in
HCO3.sup.-, Na+ and fluid re-absorption in the kidney, which is
consistently associated with hypovolemia and acidosis.
[0312] In one embodiment, the compounds of the disclosure are
intended to target the apical NHE antiporters (e.g. NHE-3, NHE-2
and NHE-8) without substantial permeability across the layer of gut
epithelial cells, and/or without substantial activity towards NHEs
that do not reside predominantly in the GI tract. This invention
provides a method to selectively inhibit GI apical NHE antiporters
and provide the desired effect of salt and fluid absorption
inhibition to correct abnormal fluid homeostasis leading to
constipations states. Because of their absence of systemic
exposure, said compounds do not interfere with other key
physiological roles of NHEs highlighted above. For instance, the
compounds of the disclosure are expected to treat constipation in
patients in need thereof, without eliciting undesired systemic
effects, such as for example salt wasting or bicarbonate loss
leading to hyponatriemia and acidosis among other disorders.
[0313] In another embodiment, the compounds of the disclosure are
delivered to the small bowel with little or no interaction with the
upper GI such as the gastric compartment and the duodenum. The
applicant found that an early release of the compounds in the
stomach or the duodenum can have an untoward effect on gastric
secretion or bicarbonate secretion (also referred to as
"bicarbonate dump"). In this embodiment the compounds are designed
so as to be released in an active form past the duodenum. This can
be accomplished by either a prodrug approach or by specific drug
delivery systems.
[0314] As used herein, "prodrug" is to be understood to refer to a
modified form of the compounds detailed herein that is inactive (or
significantly less active) in the upper GI, but once administered
is metabolised in vivo into an active metabolite after getting
past, for example, the duodenum. Thus, in a prodrug approach, the
activity of the NHE-inhibiting compound can be masked with a
transient protecting group that is liberated after compound passage
through the desired gastric compartment. For example, acylation or
alkylation of the essential guanidinyl functionality of the
NHE-inhibiting compound would render it biochemically inactive;
however, cleavage of these functional groups by intestinal
amidases, esterases, phosphatases, and the like, as well enzymes
present in the colonic flora, would liberate the active parent
compound. Prodrugs can be designed to exploit the relative
expression and localization of such phase I metabolic enzymes by
carefully optimizing the structure of the prodrug for recognition
by specific enzymes. As an example, the anti-inflammatory agent
sulfasalazine is converted to 5-aminosalicylate in the colon by
reduction of the diazo bond by intestinal bacteria.
[0315] In a drug delivery approach the NHE-inhibiting compounds of
the disclosure are formulated in certain pharmaceutical
compositions for oral administration that release the active in the
targeted areas of the GI, i.e., jejunum, ileum or colon, or
preferably the distal ileum and colon, or even more preferably the
colon.
[0316] Methods known from the skilled-in-the-art are applicable.
(See, e.g., Kumar, P. and Mishra, B., Colon Targeted Drug Delivery
Systems--An Overview, Curr. Drug Deliv., 2008, 5 (3), 186-198;
Jain, S. K. and Jain, A., Target-specific Drug Release to the
Colon., Expert Opin. Drug Deliv., 2008, 5 (5), 483-498; Yang, L.,
Biorelevant Dissolution Testing of Colon-Specific Delivery Systems
Activated by Colonic Microflora, J. Control Release, 2008, 125 (2),
77-86; Siepmann, F.; Siepmann, J.; Walther, M.; MacRae, R. J.; and
Bodmeier, R., Polymer Blends for Controlled Release Coatings, J.
Control Release 2008, 125 (1), 1-15; Patel, M.; Shah, T.; and Amin,
A., Therapeutic Opportunities in Colon-Specific Drug-Delivery
Systems, Crit. Rev. Ther. Drug Carrier Syst., 2007, 24 (2),
147-202; Jain, A.; Gupta, Y.; Jain, S. K., Perspectives of
Biodegradable Natural Polysaccharides for Site-specific Drug
Delivery to the Colon., J. Pharm. Sci., 2007, 10 (1), 86-128; Van
den, M. G., Colon Drug Delivery, Expert Opin. Drug Deliv., 2006, 3
(1), 111-125; Basit, A. W., Advances in Colonic Drug Delivery,
Drugs 2005, 65 (14), 1991-2007; Chourasia, M. K.; Jain, S. K.,
Polysaccharides for Colon-Targeted Drug Delivery, Drug Deliv. 2004,
11 (2), 129-148; Shareef, M. A.; Khar, R. K.; Ahuj a, A.; Ahmad, F.
J.; and Raghava, S., Colonic Drug Delivery: An Updated Review, AAPS
Pharm. Sci. 2003, 5 (2), E17; Chourasia, M. K.; Jain, S. K.,
Pharmaceutical Approaches to Colon Targeted Drug Delivery Systems,
J. Pharm. Sci. 2003, 6 (1), 33-66; and, Sinha, V. R.; Kumria, R.,
Colonic Drug Delivery: Prodrug Approach, Pharm. Res. 2001, 18 (5),
557-564. Typically the active pharmaceutical ingredient (API) is
contained in a tablet/capsule designed to release said API as a
function of the environment (e.g., pH, enzymatic activity,
temperature, etc.), or as a function of time. One example of this
approach is Eudracol.TM. (Pharma Polymers Business Line of
Degussa's Specialty Acrylics Business Unit), where the APL
containing core tablet is layered with various polymeric coatings
with specific dissolution profiles. The first layer ensures that
the tablet passes through the stomach intact so it can continue
through the small intestine. The change from an acidic environment
in the stomach to an alkaline environment in the small intestine
initiates the release of the protective outer layer. As it travels
through the colon, the next layer is made permeable by the
alkalinity and intestinal fluid. This allows fluid to penetrate to
the interior layer and release the active ingredient, which
diffuses from the core to the outside, where it can be absorbed by
the intestinal wall. Other methods are contemplated without
departing from the scope of the present disclosure.
[0317] In another example, the pharmaceutical compositions of the
invention can be used with drug carriers including pectin and
galactomannan, polysaccharides that are both degradable by colonic
bacterial enzymes. (See, e.g., U.S. Pat. No. 6,413,494, the entire
contents of which are incorporated herein by reference for all
relevant and consistent purposes.) While pectin or galactomannan,
if used alone as a drug carrier, are easily dissolved in simulated
gastric fluid and simulated intestinal fluid, a mixture of these
two polysaccharides prepared at a pH of about 7 or above produces a
strong, elastic, and insoluble gel that is not dissolved or
disintegrated in the simulated gastric and intestinal fluids, thus
protecting drugs coated with the mixture from being released in the
upper GI tract. When the mixture of pectin and galactomannan
arrives in the colon, it is rapidly degraded by the synergic action
of colonic bacterial enzymes. In yet another aspect, the
compositions of the invention may be used with the pharmaceutical
matrix of a complex of gelatin and an anionic polysaccharide (e.g.,
pectinate, pectate, alginate, chondroitin sulfate, polygalacturonic
acid, tragacanth gum, arabic gum, and a mixture thereof), which is
degradable by colonic enzymes (U.S. Pat. No. 6,319,518).
[0318] In yet other embodiments, fluid-absorbing polymers that are
administered in accordance with treatment methods of the present
disclosure are formulated to provide acceptable/pleasant
organoleptic properties such as mouthfeel, taste, and/or to avoid
premature swelling/gelation in the mouth and in the esophagus and
provoke choking or obstruction. The formulation may be designed in
such a way so as to ensure the full hydration and swelling of the
FAP in the GI tract and avoid the formation of lumps. The oral
dosages for the FAP may take various forms including, for example,
powder, granulates, tablets, wafer, cookie and the like, and are
most preferably delivered to the small bowel with little or no
interaction with the upper GI such as the gastric compartment and
the duodenum.
[0319] The above-described approaches or methods are only some of
the many methods reported to selectively deliver an active in the
lower part of the intestine, and therefore should not be viewed to
restrain or limit the scope of the disclosure.
[0320] The following non-limiting examples are provided to further
illustrate the present disclosure.
EXAMPLES
Exemplary Compound Synthesis
Intermediate A
(S)--N-(2-(2-(aminomethoxy)ethoxy)ethyl)-3-(6,8-dichloro-2-methyl-1,2,3,4--
tetrahydroisoquinolin-4-yl)benzenesulfonamide
##STR00018##
[0322] Intermediate A.2: (S)-tert-Butyl
(2-(2-(2-(3-(6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phe-
nylsulfonamido)ethoxy)ethoxy)ethyl)carbamate. tert-Butyl
(2-(2-(2-aminoethoxyl)ethoxy)ethyl)carbamate (8.72 g, 35.1 mmol)
was dissolved in THF (90 mL). To this solution was added a solution
of K.sub.3PO.sub.4 (37.3 g, 175 mmol) in water (90 mL). To this
rapidly stirring mixture was added solid sulfonyl chloride A.1 (see
International PCT Publication No. WO 2010/078449) (15.0 g, 35.1
mmol) in -1 g portions over 15 min. After 1 h, the mixture was
diluted with EtOAc (90 mL), and the organic layer separated. The
aqueous layer was washed with EtOAc (10 mL), and the organic
extracts combined. The solution was concentrated, and purified by
flash chromatography on silica gel eluting with a gradient of 0% to
10% methanol in DCM to give intermediate A.2 (18.7 g) as a viscous
oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.75 (d, J=7.5 Hz,
1H), 7.68 (s, 1H), 7.44 (t, J=7.7 Hz, 1H), 7.36 (d, J=7.4 Hz, 1H),
7.26-7.21 (m, 1H), 6.73 (s, 1H), 5.28 (s, 1H), 5.08 (s, 1H), 4.25
(t, J=6.1 Hz, 1H), 3.79-3.44 (m, 10H), 3.30 (d, J=4.5 Hz, 2H), 3.12
(t, J=4.6 Hz, 2H), 2.93 (dd, J=11.5, 5.2 Hz, 1H), 2.58 (dd, J=11.6,
7.2 Hz, 1H), 2.44 (s, 3H), 1.42 (s, 9H). Mass (ESI+) 602.10
(M+H.sup.+).
[0323] Intermediate A:
(S)--N-(2-(2-(aminomethoxy)ethoxy)ethyl)-3-(6,8-dichloro-2-methyl-1,2,3,4-
-tetrahydroisoquinolin-4-yl)benzenesulfonamide. Intermediate A.2
(15.2 g, 25.2 mmol) was dissolved in warm (45.degree. C.)
2-propanol (80 mL). This solution was added to aqueous 25%
H.sub.2SO.sub.4 at 45.degree. C., over 1 h. The solution was
stirred for an additional hour, then concentrated under reduced
pressure to remove the 2-propanol. DCM (150 mL) was added, and the
pH of the mixture was adjusted with K.sub.3PO.sub.4 to pH 7-8 using
pH paper. The DCM layer was separated, and the aqueous layer was
extracted three more times with DCM (150 mL). The organic layer was
dried, and concentrated under reduced pressure to give intermediate
A (12 g) as a foam. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.83
(d, J=7.9 Hz, 1H), 7.77 (s, 1H), 7.41 (t, J=7.4 Hz, 1H), 7.31 (d,
J=7.4 Hz, 1H), 7.22 (s, 1H), 6.72 (s, 1H), 4.26 (t, J=5.3 Hz, 1H),
3.79-3.47 (m, 12H), 3.13 (d, J=32.2 Hz, 4H), 2.93 (dd, J=11.8, 5.1
Hz, 1H), 2.59 (dd, J=10.7, 7.4 Hz, 1H), 2.43 (s, 3H). Mass (ESI+)
502.11 (M+H.sup.+).
Intermediate B
bis(perfluorophenyl)
4-nitro-4-(3-oxo-3-(perfluorophenoxy)propyl)heptanedioate
##STR00019##
[0325] Intermediate B: A solution of
4-(2-carboxyethyl)-4-nitroheptanedioic acid (3.00 g, 10.8 mmol) in
DCM (54 mL) was charged in an additional funnel and added dropwise
to a solution of perfluorophenyl 2,2,2-trifluoroacetate (6.15 mL,
35.7 mmol) and TEA (9.0 mL, 65 mmol) in DCM (54 mL). Upon
completion of addition, the solution was stirred an additional 20
min at room temperature, during which time a white precipitate
formed. The precipitate was filtered and washed with 3:7
DCM:hexanes and then washed with hexanes to give the title compound
(6.87 g, 82%) as a white solid. .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta. 2.88-2.71 (m, 6H), 2.59-2.41 (m, 6H). .sup.19F-NMR (376
MHz, CDCl.sub.3) .delta. -152.71 (d, J=17.1 Hz), -157.08 (t, J=21.7
Hz), -161.86 (dt, J=21.4, 10.7 Hz).
Intermediate C
tris(perfluorophenyl) 2,2',2''-nitrilotriacetate
##STR00020##
[0327] Intermediate C: The title compound was synthesized in a
manner similar to bis(perfluorophenyl)
4-nitro-4-(3-oxo-3-(perfluorophenoxy)propyl)heptanedioate, using
2,2',2''-nitrilotriacetic acid in place of
4-(2-carboxyethyl)-4-nitroheptanedioic acid.
Intermediate D
(9H-fluoren-9-yl)methyl
1,25-bis(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)-
phenylsulfonamido)-13-(3-(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-te-
trahydroisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethylamino)-3-oxo-
propyl)-10,16-dioxo-3,6,20,23-tetraoxa-9,17-diazapentacosan-13-ylcarbamate
##STR00021##
[0329] Intermediate D:
4-(((9H-fluoren-9-yl)methoxy)carbonylamino)-4-(2-carboxyethyl)heptanedioi-
c acid (2.07 g, 4.40 mmol) was dissolved in THF (80 mL), then
carbonyldiimidazole (2.21 g, 13.6 mmol) was added and the resulting
mixture stirred for 2 h at room temperature. Additional
carbonyldiimidazole (357 mg, 2.20 mmol) was the added in three
portions over a period of 1.5 h, until a sample of the reaction
mixture quenched with N.sup.1,N.sup.1-dimethylpropane-1,3-diamine
showed all of the starting material was consumed. To the mixture
was then added a solution of A (7.30 g, 14.5 mmol) in DMF and the
resulting solution stirred for 2 h at room temperature. The
solution was then poured into H.sub.2O (800 mL) and the resulting
white precipitate filtered. The precipitate was then dissolved in
DCM and washed with 1 M aqueous HCl and saturated aqueous
NaHCO.sub.3, then the solvent removed under reduced pressure to
give D (8.36 g, 99%) as a light yellow foam. .sup.1H-NMR (400 MHz,
CDCl.sub.3) .delta. 7.76-7.67 (m, 7H), 7.57 (t, J=8.1 Hz, 3H), 7.39
(t, J=7.7 Hz, 3H), 7.37-7.27 (m, 6H), 7.26-7.18 (m, 2H), 6.73-6.63
(m, 5H), 4.30-4.19 (m, 6H), 4.05 (t, J=6.7 Hz, 1H), 3.61 (dd,
J=36.9, 16.2 Hz, 6H), 3.54-3.42 (m, 21H), 3.37 (dd, J=10.1, 4.9 Hz,
6H), 3.11-3.01 (m, 6H), 2.95-2.84 (m, 5H), 2.55 (dd, J=11.5, 7.2
Hz, 3H), 2.41 (s, 9H), 2.28-2.17 (m, 6H), 2.04-1.93 (m, 6H). MS
(ES, m/z): 1919.3 [M+H].sup.+.
Intermediate E
(S)--N-(2-(2-(2-(2-aminoethoxyl)ethoxy)ethoxy)ethyl)-3-(6,8-dichloro-2-met-
hyl-1,2,3,4-tetrahydroisoquinolin-4-yl)benzenesulfonamide
##STR00022##
[0331] Intermediate E.1:
(S)--N-(2-(2-(2-(2-azidoethoxyl)ethoxy)ethoxy)ethyl)-3-(6,8-dichloro-2-me-
thyl-1,2,3,4-tetrahydroisoquinolin-4-yl)benzenesulfonamide.
2-(2-(2-(2-azidoethoxyl)ethoxy)ethoxy)ethanamine (6.49, 29.7 mmol)
was dissolved in THF (90 mL). To this solution was added a solution
of K.sub.3PO.sub.4 (27.5 g, 130 mmol) in water (90 mL). To this
rapidly stirring mixture was added solid sulfonyl chloride A.1
(12.1 g, 28.3 mmol) in -1 g portions over 15 min. After 1 h, the
mixture was diluted with EtOAc (90 mL), and the organic layer
separated. The aqueous layer was washed with EtOAc (10 mL), and the
organic extracts combined. The solution was concentrated, and
purified by flash chromatography on silica eluting with a gradient
of 0% to 10% methanol in DCM to give intermediate E.1 (16 g) as a
viscous oil. .sup.1H NMR (400 MHz, CDCl.sub.3) 7.73 (d, J=7.9 Hz,
1H), 7.67 (s, 1H), 7.43 (t, J=7.9 Hz, 1H), 7.35 (d, J=7.6 Hz, 1H),
6.72 (s, 1H), 5.23 (dd, J=6.1, 5.7 Hz, 1H), 4.23-4.25 (m, 1H),
3.67-3.47 (comp, 14H), 3.37 (t, J=4.9 Hz, 2H), 3.11 (q, J=10.4, 5.5
Hz, 2H), 2.93 (dd, J=5.2, 11.6 Hz, 2H), 2.58 (dd, J=11.7, 7.4 Hz,
1H), 2.43 (s, 3H). Mass (ESI+) 572.12 (M+H.sup.+)
[0332] Intermediate E:
(S)--N-(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-3-(6,8-dichloro-2-met-
hyl-1,2,3,4-tetrahydroisoquinolin-4-yl)benzenesulfonamide. To a
solution of Intermediate E.1 (13.0 g, 22.7 mmol) in THF (75 mL) at
10.degree. C., was added trimethylphosphine (3.46 g, 45.4 mmol)
keeping the internal temperature under 15.degree. C. The solution
was stirred for 1 h at 10.degree. C., then warmed to 20.degree. C.
for 1 hr. An aqueous solution of ice cold NaOH (1M, 10 mmol, 10 mL)
was added, then after 15 min, the mixture was concentrated under
reduced pressure to remove the bulk of the THF. The stirring
aqueous mixture was diluted with DCM (1.5 L), and water (100 mL)
was added, followed by aqueous 25% NaCl solution (25 mL). Agitation
was stopped, and the mixture separated (.about.1 h). The aqueous
layer was extracted twice with DCM (200 mL), and the combined
extracts were dried (MgSO.sub.4), and concentrated under reduced
pressure to give intermediate E (13.5 g) as a tacky foam. 1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.78-7.71 (m, 1H), 7.67 (t, J=1.7 Hz,
1H), 7.42 (t, J=7.7 Hz, 1H), 7.33 (d, J=7.8 Hz, 1H), 7.26-7.21 (m,
1H), 6.73 (s, 1H), 4.25 (t, J=6.3 Hz, 1H), 3.76-3.46 (m, 8H),
3.14-3.08 (m, 2H), 2.93 (dd, J=11.7, 5.6 Hz, 1H), 2.84 (t, 2H),
2.58 (dd, J=11.7, 7.4 Hz, 1H), 2.44 (s, 3H). Mass (ESI+) 546.12
(M+H.sup.+).
Intermediate F
Tris(perfluorophenyl) 3,3',3''-nitrilotripropanoate
##STR00023##
[0334] Intermediate F: The title compound was synthesized in a
manner similar to bis(perfluorophenyl)
4-nitro-4-(3-oxo-3-(perfluorophenoxy)propyl)heptanedioate, using
3,3',3''-nitrilotripropanoic acid in place of
4-(2-carboxyethyl)-4-nitroheptanedioic acid.
Intermediate G
N.sup.1,N.sup.7-bis(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahyd-
roisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethyl)-4-(3-(2-(2-(2-(3-
-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfo-
namido)ethoxy)ethoxy)ethylamino)-3-oxopropyl)-4-isocyanatoheptanediamide
##STR00024##
[0336] Intermediate G: To a solution of example 2 (120 mg, 0.0706
mmol) and triethylamine (49 .mu.L, 0.35 mmol) in DCM (2 mL) and THF
(2 mL) cooled to 0.degree. C. was added triphosgene (10.5 mg, 0.353
mmol). The solution was then allowed to warm to room temperature
and stirred for 30 min, then stirred an additional 1 h at
40.degree. C. The mixture was then diluted with DCM and washed with
saturated aqueous NaHCO.sub.3. The organic layer dried over
Na.sub.2SO4 and then the solvent removed under reduces pressure to
give the product as a yellow oil, which was used directly without
further purification. MS (ES, m/z): 1722.8 [M+H].sup.+.
Intermediate H
(S)-3-(6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)-N-(2-(2-(-
2-(methylamino)ethoxy)ethoxy)ethyl)benzenesulfonamide
##STR00025##
[0338] Intermediate H.1: tert-butyl
(2-(2-(2-aminoethoxyl)ethoxy)ethyl)carbamate, To a mixture of 2,
2'-(ethylenedioxy)bis(ethylamine) (153 g, 1.032 mol, 4.5 equiv) in
DCM (250 mL) at 0.degree. C. was added a solution of di-tert-butyl
dicarbonate (50 g, 0.229 mol, 1 equiv) in DCM (100 mL) over 3.5 h.
The mixture was slowly warmed to rt, stirred at rt overnight and
washed with 25% aqueous NaCl solution (3.times.100 mL) and water
(100 mL). The organic layer was extracted with 20% aqueous citric
acid (250 mL). The aqueous layer was washed with DCM (150 mL),
basified to pH 13-14 by aqueous NaOH solution (2 M), extracted with
DCM (3.times.). The combined organic layers were dried and
concentrated to give 47 g (83%) of intermediate H.1 as clear
oil.
[0339] Intermediate H.2: tert-butyl
(2-(2-(2-(2-nitrophenylsulfonamido)ethoxy)ethoxy)ethyl)carbamate,
To a mixture of tert-butyl
(2-(2-(2-aminoethoxyl)ethoxy)ethyl)carbamate (4.0 g, 16.15 mmol,
1.0 equiv) and triethylamine (3.38 mL, 24.22 mmol, 1.5 equiv) in
DCM (30 mL) at 0.degree. C. was added a solution of
2-nitrobenzenesulfonyl chloride (3.76 g, 16.95 mmol, 1.05 equiv) in
DCM (20 mL) dropwise. The mixture was stirred at rt overnight,
diluted with ethyl acetate and washed with 10% citric acid
(1.times.), H.sub.2O (1.times.), sat. aqueous NaHCO.sub.3
(1.times.), brine (1.times.). The organic layer was dried and
concentrated to give 7.34 g of intermediate H.2 as yellow
syrup.
[0340] Intermediate H.3: tert-butyl
(2-(2-(2-(N-methyl-2-nitrophenylsulfonamido)ethoxy)ethoxy)ethyl)carbamate-
. To a mixture of tert-butyl
(2-(2-(2-(2-nitrophenylsulfonamido)ethoxy)ethoxy)ethyl)carbamate
(7.34 g, 16.96 mmol, 1.0 equiv) in DMF (50 mL) were added
K.sub.2CO.sub.3 (3.51 g, 25.44 mmol, 1.5 equiv) and iodomethane
(1.48 mL, 23.74 mmol, 1.4 equiv). The mixture was stirred at rt for
1.5 h, diluted with ethyl acetate, washed with H.sub.2O (2.times.)
and brine (1.times.), dried, and concentrated to give 7.58 g of
intermediate H.3 as yellow syrup.
[0341] Intermediate H.4:
N-(2-(2-(2-aminoethoxyl)ethoxy)ethyl)-N-methyl-2-nitrobenzenesulfonamide.
To a mixture of tert-butyl
(2-(2-(2-(N-methyl-2-nitrophenylsulfonamido)ethoxy)ethoxy)ethyl)carbamate
(7.58 g) in DCM (2 mL) was added a solution of HCl in dioxane (4 M,
40 mL). The mixture was stirred at rt for 40 minutes and
concentrated to give 7.3 g of intermediate H.4 HCl salt as yellow
syrup.
[0342] Intermediate H.5:
(S)--N-(2-(2-(2-(3-(6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-
-yl)-phenylsulfonamido)ethoxy)ethoxy)ethyl)-N-methyl-2-nitrobenzenesulfona-
mide. To a mixture of
N-(2-(2-(2-aminoethoxyl)ethoxy)ethyl)-N-methyl-2-nitrobenzenesulfonamide
(7.3 g crude, about 16.15 mmol, 1 equiv) and TEA (11.25 mL, 80.73
mmol, 5 equiv) in DCM (80 mL) at 0.degree. C. was added
(S)-3-(6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)benzene-1-
-sulfonyl chloride A.1 (7.24 g, 16.95 mmol, 1.05 equiv). The
mixture was stirred at rt for 1 h, diluted with ethyl acetate,
washed with water (1.times.) and brine (1.times.), dried,
concentrated, and purified by column to give 9.84 g (87%, 4 steps)
of intermediate H.5 as a yellow solid.
[0343] Intermediate H:
(S)-3-(6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)-N-(2-(2--
(2-(methylamino)ethoxy)ethoxy)ethyl)benzenesulfonamide. To a
mixture of
(S)--N-(2-(2-(2-(3-(6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-
-yl)-phenylsulfonamido)ethoxy)ethoxy)ethyl)-N-methyl-2-nitrobenzenesulfona-
mide (5.1 g, 7.28 mmol, 1 equiv) and K.sub.2CO.sub.3 (3.01 g, 21.83
mmol, 3 equiv) in DMF (30 mL) at rt was added thiophenol (1.12 mL,
10.91 mmol, 1.5 equiv). The mixture was stirred at rt for 1 h,
diluted with ether and extracted with IN aqueous HCl. The aqueous
layer was washed with ether (2.times.), basified with NaHCO.sub.3
to pH 9, and extracted with DCM (3.times.). The combined organic
layers were dried, concentrated and purified by C-18 column to give
4.03 g (75%) of the title compound TFA salt as a white solid. MS
(ES, m/z): 516 [M+H].sup.+. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.77 (dt, J=7.6, 1.5 Hz, 1H), 7.70-7.65 (m, 1H), 7.58-7.47
(m, 2H), 7.35 (d, J=1.5 Hz, 1H), 6.80 (d, J=1.2 Hz, 1H), 4.40 (t,
J=6.5 Hz, 1H), 3.78 (d, J=16.2 Hz, 1H), 3.70-3.62 (m, 3H),
3.62-3.58 (m, 2H), 3.57-3.53 (m, 2H), 3.51-3.46 (m, 2H), 3.05-2.99
(m, 3H), 2.98-2.91 (m, 2H), 2.67 (dd, J=11.7, 7.8 Hz, 1H), 2.55 (s,
3H), 2.48 (s, 3H).
Intermediate I
(S)--N-(2-(2-(2-aminoethoxyl)ethoxy)ethyl)-3-(6,8-dichloro-2-methyl-1,2,3,-
4-tetrahydroisoquinolin-4-yl)-N-methylbenzenesulfonamide
##STR00026##
[0345] Intermediate I.1: (S)-t-butyl
(2-(2-(2-(3-(6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)-N--
methylphenylsulfonamido)ethoxy)ethoxy)ethyl)carbamate. To a mixture
of (S)-t-butyl
(2-(2-(2-(3-(6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phe-
nylsulfonamido)ethoxy)ethoxy)ethyl)carbamate (352 mg, 0.586 mmol, 1
equiv), methanol (47.4 .mu.L, 1.17 mmol, 2 equiv) and PPh.sub.3
(307 mg, 1.17 mmol, 2 equiv) in THF (2 mL) at 0.degree. C. was
added dropwise a solution of diethyl azodicarboxylate (40% in
toluene, 0.534 mL, 1.17 mmol, 2 equiv). The mixture was stirred at
rt overnight, concentrated and purified by column to give 0.9 g
(crude) of intermediate I.1 as yellow syrup.
[0346] Intermediate I:
(S)--N-(2-(2-(2-aminoethoxyl)ethoxy)ethyl)-3-(6,8-dichloro-2-methyl-1,2,3-
,4-tetrahydroisoquinolin-4-yl)-Nmethylbenzenesulfonamide. To a
mixture of (S)-t-butyl
(2-(2-(2-(3-(6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)-N--
methylphenylsulfonamido)ethoxy)ethoxy)ethyl)carbamate (0.7 g) in
DCM (0.5 mL) was added a solution of HCl in dioxane 4 M, 3 mL). The
mixture was stirred at rt for 0.5 h, concentrated and purified by
prep HPLC to give 200 mg (59%, 2 steps) of intermediate I as a
white solid. MS (ES, m/z): 516 [M+H].sup.+. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 7.87-7.80 (m, 1H), 7.73-7.64 (m, 2H), 7.60 (d,
J=7.8 Hz, 1H), 7.55 (d, J=2.0 Hz, 1H), 6.83 (s, 1H), 4.84-4.73 (m,
2H), 4.50 (d, J=16.0 Hz, 1H), 3.90 (ddd, J=12.2, 5.9, 1.3 Hz, 1H),
3.75-3.70 (m, 2H), 3.69-3.62 (m, 6H), 3.59 (d, J=12.0 Hz, 1H), 3.21
(dd, J=9.3, 5.2 Hz, 2H), 3.17-3.08 (m, 5H), 2.80 (s, 3H).
Intermediate J
N.sup.1-methyl-N.sup.3,N.sup.3-bis(3-(methylamino)propyl)propan-1,3-diamin-
e
##STR00027##
[0348] Intermediate J:
N.sup.1-methyl-N.sup.3,N.sup.3-bis(3-(methylamino)propyl)propan-1,3-diami-
ne To a mixture of
tri-t-butyl(nitrilotris(propane-3,1-diyl))tricarbamate (689 mg,
1.41 mmol, 1 equiv) in THF (8 mL) 0.degree. C. was added lithium
aluminium hydride (2M in THF, 4.24 mL, 8.48 mmol, 6 equiv). The
mixture was slowly warmed to 70.degree. C. and stirred at
70.degree. C. for 3 h. The reaction was carefully quenched with
Na.sub.2SO.sub.4.10H.sub.2O and filtered. The filtrate was
concentrated to give 281 mg (87%) of intermediate J as clear syrup.
MS (ES, m/z): 231 [M+H].sup.+. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 2.57 (t, J=6.8 Hz, 6H), 2.47-2.34 (m, 15H), 1.62 (dt,
J=14.0, 7.0 Hz, 6H).
Intermediate K
(S)--N-(2-(2-(2-aminoethoxyl)ethoxy)ethyl)-3-(6,8-dichloro-2-methyl-1,2,3,-
4-tetrahydroisoquinolin-4-yl)-N-(2-hydroxyethyl)benzenesulfonamide
##STR00028##
[0350] Intermediate K:
(S)--N-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-3-(6,8-dichloro-2-methyl-1,2,3,-
4-tetrahydroisoquinolin-4-yl)-N-(2-hydroxyethyl)benzenesulfonamide.
Intermediate K was synthesized in an analogous fashion to
intermediate I, using 2-(tert-butoxy)ethanol in place of methanol.
MS (ES, m/z): 546 [M+H].sup.+.
Intermediate L
(S)--N-(2-(2-(2-aminoethoxyl)ethoxy)ethyl)-3-(6,8-dichloro-2-methyl-1,2,3,-
4-tetrahydroisoquinolin-4-yl)-N-ethylbenzenesulfonamide
##STR00029##
[0352] Intermediate L:
(S)--N-(2-(2-(2-aminoethoxyl)ethoxy)ethyl)-3-(6,8-dichloro-2-methyl-1,2,3-
,4-tetrahydroisoquinolin-4-yl)-N-ethylbenzenesulfonamide.
Intermediate L was synthesized in an analogous fashion to
intermediate I, using ethanol in place of methanol. MS (ES, m/z):
530 [M+H].sup.+.
Intermediate M
3-amino-N,N-bis(3-aminopropyl)-N-methylpropan-1-aminium
##STR00030##
[0354] Intermediate M.1:
tri-t-butyl(nitrilotris(propane-3,1-diyl))tricarbamate. To a
mixture of N.sup.1,N.sup.1-bis(3-aminopropyl)propane-1,3-diamine
(499 mg, 2.65 mmol, 1 equiv) in DCM (10 mL) at 0.degree. C. were
added di-tert-butyl dicarbonate (2.08 g, 9.54 mol, 3.6 equiv) and
TEA (1.66 mL, 11.92 mmol, 4.5 equiv). The mixture was slowly warmed
to rt, stirred at rt overnight, diluted with ethyl acetate, washed
with water (1.times.) and brine (1.times.), dried and concentrated
to give 1.4 g (crude) of intermediate M.1.
[0355] Intermediate M.2:
3-((t-butoxycarbonyl)amino)-N,N-bis(3-((t-butoxycarbonyl)amino)propyl)-N--
methylpropan-1-aminium. To a mixture of
tri-t-butyl(nitrilotris(propane-3,1-diyl))tricarbamate (258.2 mg,
0.529 mmol, 1 equiv) in acetonitrile (5 mL) at rt was added
iodomethane (39.6 .mu.L, 0.635 mol, 1.2 equiv). The mixture was
stirred at rt overnight and concentrated to give 286 mg (86%) of
intermediate M.2 as clear syrup.
[0356] Intermediate M:
3-amino-N,N-bis(3-aminopropyl)-N-methylpropan-1-aminium. To a
mixture of
3-((t-butoxycarbonyl)amino)-N,N-bis(3-((t-butoxycarbonyl)amino)propyl)-N--
methylpropan-1-aminium (286 mg) in DCM (0.5 mL) was added a
solution of HCl in dioxane (4 M, 3 mL). The mixture was stirred at
rt for 1 h and concentrated to give 190 mg (crude) of intermediate
M as a yellow solid. MS (ES, m/z): 203 [M].sup.+. .sup.1H NMR (400
MHz, CD.sub.3OD) .delta. 3.62-3.54 (m, 6H), 3.22 (s, 3H), 3.11 (t,
J=7.4 Hz, 6H), 2.30-2.19 (m, 6H).
Intermediate N
4-acetyl-4-(2-carboxyethyl)heptanedioic acid
##STR00031##
[0358] Intermediate N.1 To a well-stirred solution of acetone (300
mg, 5.15 mmol) and 30% ethanolic KOH (25 .mu.L) in t-BuOH (0.30 mL)
at 0.degree. C. was added a solution of acrylonitrile (0.82 g, 15.5
mmol) in t-BuOH (0.40 mL) over 1 hour. The reaction mixture was
then stored at 4.degree. C. overnight. The solids were collected on
a Buchner funnel, and washed with water (2.times.5 mL). The product
was dissolved in acetonitrile (10 mL) and DCM (50 mL), dried
(Na.sub.2SO.sub.4) and concentrated to give N.1 (667 mg) as an
off-white solid.
[0359] Intermediate N: A mixture of
4-acetyl-4-(2-cyanoethyl)heptanedinitrile (667 mg, 3.1 mmol) and
KOH, 840 g, 15 mmol) in water (4.8 mL) was heated at reflux for 5
hours. The reaction mixture was cooled to 50.degree. C. and
decanted from insoluble gum. The supernatant was acidified to
pH-2-3 with Conc. HCl and concentrated to dryness under vacuum. The
semisolid residue was heated at 50.degree. C. with acetone (20 mL)
and the mixture was filtered hot, and concentrated to give
intermediate N as an oil (690 mg) which crystalized on seeding with
crystals generated from a small aliquot in DCM.
Example 1
N1,N7-bis(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquino-
lin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethyl)-4-(3-(2-(2-(2-(3-((S)-6,8--
dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfonamido)eth-
oxy)ethoxy)ethylamino)-3-oxopropyl)-4-nitroheptanediamide
##STR00032##
[0361] Example 1: To a solution of A (972 mg, 193 mmol) and DIEA
(657 .mu.L, 3.87 mmol) in DCM (20 mL) was added
bis(perfluorophenyl)
4-nitro-4-(3-oxo-3-(perfluorophenoxy)propyl)heptanedioate
(intermediate B, 500 mg, 0.645 mmol) and the resulting solution
stirred at room temperature for 20 h. The solvent was removed under
reduced pressure and the resulting residue purified by automated
flash column silica gel chromatography using a gradient of DCM:MeOH
(99:1 to 9:1) to give the title compound as a yellow solid (516 mg,
46%) after the solvent was removed. .sup.1H-NMR (400 MHz,
CD.sub.3OD) .delta. 7.79-7.75 (m, 3H), 7.70 (t, J=1.5 Hz, 3H), 7.53
(t, J=7.6 Hz, 3H), 7.50-7.45 (m, 3H), 7.34 (d, J=2.1 Hz, 3H), 6.80
(s, 3H), 4.44-4.36 (m, 3H), 3.77 (d, J=16.1 Hz, 3H), 3.64 (d,
J=15.6 Hz, 3H), 3.57-3.48 (m, 18H), 3.45 (t, J=5.5 Hz, 6H), 3.34 (,
J=5.2 Hz, 6H), 3.07-2.99 (m, 9H), 2.67 (dd, J=11.7, 7.8 Hz, 3H),
2.47 (s, 9H), 2.28-2.16 (m, 12H). MS (ES, m/z): 1727.1
[M+H].sup.+.
Example 2
4-amino-N.sup.1,N.sup.7-bis(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4--
tetrahydroisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethyl)-4-(3-(2--
(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phe-
nylsulfonamido)ethoxy)ethoxy)ethylamino)-3-oxopropyl)heptanediamide
##STR00033##
[0363] Example 2, method A: To a solution of
N.sup.1,N.sup.1-bis(2-aminoethyl)ethane-1,2-diamine (1.21 mL, 8.06
mmol) in DMF (2 mL) was slowly added a solution of intermediate D
(7.75 g, 4.03 mmol) in DMF (10 mL) and the resulting mixture
stirred for 30 min at room temperature. The solution was cooled to
0.degree. C. and 1 M aqueous TFA was added (20 mL), until the
solution reached pH=1. The solution was then diluted with 1:1
MeCN:H.sub.2O to give a final volume of 60 mL. The solution was
purified by automated flash column reverse phase chromatography
using a gradient of H.sub.2O 0.05% TFA: CH.sub.3CN 0.05% TFA (80:20
to 60:40) and detection by UV at 254 nm in three batches. The
fractions containing pure material were concentrated and then
neutralized to pH=7 with NaHCO.sub.3, resulting in the formation of
a white precipitate. The suspension was extracted twice with a 95:5
DCM: MeOH solution. The combined organic layers were dried over
Na.sub.2SO.sub.4 and the solvent removed to give the title compound
(3.52 g, 51% yield) as a white foam. .sup.1H-NMR (400 MHz,
CD.sub.3OD) .delta. 7.89 (d, J=7.9 Hz, 3H), 7.77 (t, J=1.6 Hz, 3H),
7.65 (t, J=7.8 Hz, 3H), 7.58-7.52 (m, 6H), 6.83 (s, 3H), 4.81-4.71
(m, 6H), 4.47 (d, J=15.9 Hz, 3H), 3.87 (dd, J=12.4, 6.0 Hz, 3H),
3.64-3.51 (m, 21H), 3.48 (t, J=5.4 Hz, 6H), 3.36 (t, J=5.5 Hz, 6H),
3.13 (s, 9H), 3.05 (t, J=5.4 Hz, 6H), 2.44-2.34 (m, 6H), 1.97 (m,
6H). MS (ES, m/z): 1697.2 [M+H].sup.+.
##STR00034##
[0364] Example 2, method B: To a Parr hydrogenation bottle was
added example 1 (926 mg, 0.535 mmol) in MeOH (40 mL) and Raney.RTM.
nickel (1.0 g), which had been washed five times with H.sub.2O,
until the aqueous layer pH=7. The bottle was shaken for 16 h at
room temperature under 50 psi of H.sub.2. Additional washed
Raney.RTM. nickel (1.0 g) was then added and the suspension shaken
for 16 h under 50 psi of H.sub.2. A final addition of washed
Raney.RTM. nickel (2.0 g) was then added and shaken for 16 h under
50 psi of H.sub.2, at which time analysis by LCMS showed all
starting material has been consumed. The suspension was filtered
through a pad of Celite.RTM. and the pad washed twice with EtOH. To
combined organic layers were concentrated under reduced pressure
and was purified by automated flash column reverse phase
chromatography using a gradient of H.sub.2O 0.05% TFA: CH.sub.3CN
0.05% TFA (80:20 to 50:50) and detection by UV at 254 nm. The
solvent was removed under reduced pressure and the resulting
residue dissolved in DCM and washed with saturated aqueous
NaHCO.sub.3. The organic phase was dried over Na.sub.2SO.sub.4 and
the solvent removed under reduced pressure to give the title
compound (280 mg, 31%).
Example 3
1-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenyls-
ulfonamido)-13,13-bis(3-(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tet-
rahydroisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethylamino)-3-oxop-
ropyl)-10-oxo-3,6-dioxa-9,14-diazahexadecane-16-sulfonic acid
##STR00035##
[0366] Example 3: Taurine (9.2 mg, 0.074 mmol) was dissolved in
H.sub.2O (200 .mu.L), to which was added DIEA (26 .mu.L, 0.15
mmol), followed by DMF (800 .mu.L). To the resulting solution was
added N,N'-disuccinimidyl carbonate (19 mg, 0.074 mmol) and the
solution stirred at 50.degree. C. for 1. Example 2 (25 mg, 0.015
mmol) was then added and the solution stirred for 18 h at
50.degree. C. The solution was then diluted with H.sub.2O and
acidified with TFA, then purified by preparative HPLC with a C18
silica gel stationary phase using a gradient of H.sub.2O 0.05%
TFA:CH.sub.3CN 0.05% TFA (80:20 to 40:60) and detection by UV at
254 nm to give the title compound tri-TFA salt (10 mg, 30% yield)
as a white solid. .sup.1H-NMR (400 MHz, CD.sub.3OD) .delta.
7.91-7.86 (m, 3H), 7.86-7.82 (m, 3H), 7.64 (t, J=7.8 Hz, 3H),
7.58-7.51 (m, 6H), 6.84 (s, 3H), 4.83-4.74 (m, 6H), 4.54 (d, J=15.4
Hz, 3H), 3.93 (dd, J=12.0, 6.2 Hz, 3H), 3.66 (t, J=11.9 Hz, 3H),
3.58-3.49 (m, 18H), 3.47 (t, J=5.4 Hz, 6H), 3.44-3.37 (m, 2H),
3.35-3.32 (m, 6H), 3.17 (d, J=9.0 Hz, 9H), 3.06 (t, J=5.3 Hz, 6H),
2.90 (t, J=6.2 Hz, 2H), 2.24-2.13 (m, 6H), 1.92-1.81 (m, 6H). MS
(ES, m/z): 1847.9 [M+H].sup.+.
Example 4
N.sup.1,N.sup.7-bis(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahyd-
roisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethyl)-4-(3-(2-(2-(2-(3-
-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfo-
namido)ethoxy)ethoxy)ethylamino)-3-oxopropyl)-4-(dimethylamino)heptanediam-
ide
##STR00036##
[0368] Example 4: Example 2 (200 mg, 0.118 mmol) and 33 weight %
aqueous formaldehyde (30 .mu.L) were combined in a mixture of MeCN
(2 mL) and H.sub.2O (2 mL). Five drops of acetic acid were then
added, followed by sodium triacetoxyborohydride (15 mg, 0.24 mmol)
and the mixture stirred for 30 min at room temperature. The mixture
was then purified by preparative HPLC with a C18 silica gel
stationary phase using a gradient of H.sub.2O 0.05% TFA:CH.sub.3CN
0.05% TFA (80:20 to 40:60) and detection by UV at 254 nm to give
the title compound tetra-TFA salt (146 mg, 57% yield) as a white
solid. .sup.1H-NMR (400 MHz, CD.sub.3OD) .delta. 7.89 (d, J=7.9 Hz,
3H), 7.77 (s, 3H), 7.66 (t, J=7.8 Hz, 3H), 7.59-7.51 (m, 6H), 6.83
(s, 3H), 4.82-4.74 (m, 6H), 4.50 (d, J=15.9 Hz, 3H), 3.90 (dd,
J=11.8, 6.3 Hz, 3H), 3.67-3.51 (m, 21H), 3.48 (t, J=5.4 Hz, 6H),
3.37 (t, J=5.4 Hz, 6H), 3.16 (s, 9H), 3.05 (t, J=5.4 Hz, 6H), 2.91
(s, 6H), 2.50-2.37 (m, 6H), 2.16-2.06 (m, 6H). MS (ES, m/z): 1725.0
[M+H].sup.+.
Example 5
N.sup.1,N.sup.7-bis(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahyd-
roisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethyl)-4-(3-(2-(2-(2-(3-
-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfo-
namido)ethoxy)ethoxy)ethylamino)-3-oxopropyl)-4-(methylsulfonamido)heptane-
diamide
##STR00037##
[0370] Example 5: Example 2 (47 mg, 0.028 mmol) and DIEA (14 .mu.L,
0.83 mmol) were dissolved in MeCN (1 mL). Methanesulfonic anhydride
(6.0 mg, 0.35 mmol) was then added and the solution stirred for 1 h
at room temperature and then stirred for an additional 1 h at
50.degree. C. The solution was then diluted with H.sub.2O and
acidified with TFA, then purified by preparative HPLC with a C18
silica gel stationary phase using a gradient of H.sub.2O 0.05%
TFA:CH.sub.3CN 0.05% TFA (80:20 to 20:80) and detection by UV at
254 nm to give the title compound tri-TFA salt (6.7 mg, 11% yield)
as a white solid. .sup.1H-NMR (400 MHz, CD.sub.3OD) .delta. 7.89
(d, J=8.1 Hz, 3H), 7.78 (s, 3H), 7.65 (t, J=7.8 Hz, 3H), 7.58-7.52
(m, 6H), 6.84 (s, 3H), 4.81-4.72 (m, 6H), 4.51 (d, J=15.6 Hz, 3H),
3.91 (dd, J=12.0, 6.0 Hz, 3H), 3.63 (t, J=12.1 Hz, 3H), 3.58-3.50
(m, 18H), 3.48 (t, J=5.4 Hz, 6H), 3.34 (t, J=5.5 Hz, 6H), 3.17 (s,
9H), 3.09-3.02 (m, 9H), 2.34-2.24 (m, 6H), 1.96-1.86 (m, 6H). MS
(ES, m/z): 1775.1 [M+H].sup.+.
Example 6
1-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenyls-
ulfonamido)-13,13-bis(3-(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tet-
rahydroisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethylamino)-3-oxop-
ropyl)-10,15-dioxo-3,6,17-trioxa-9,14-diazanonadecan-19-oic
acid
##STR00038##
[0372] Example 6: Example 2 (50 mg, 0.029 mmol) and DIEA (15 .mu.L,
0.088 mmol) were dissolved in DMF (1 mL). 1,4-Dioxane-2,6-dione
(6.0 mg, 0.038 mmol) was then added and the solution stirred at
40.degree. C. for 1 h, then diluted with H.sub.2O and acidified
with TFA. The mixture was then purified by preparative HPLC with a
C18 silica gel stationary phase using a gradient of H.sub.2O 0.05%
TFA: CH.sub.3CN 0.05% TFA (80:20 to 20:80) and detection by UV at
254 nm to give the title compound tri-TFA salt (40 mg, 63% yield)
as a white solid. .sup.1H-NMR (400 MHz, CD.sub.3OD) .delta.
7.91-7.86 (m, 3H), 7.79 (t, J=1.6 Hz, 3H), 7.64 (t, J=7.8 Hz, 3H),
7.58-7.52 (m, 6H), 6.84 (s, 3H), 4.84-4.75 (m, 6H), 4.52 (d, J=15.9
Hz, 3H), 4.17 (s, 2H), 3.99 (s, 2H), 3.92 (dd, J=11.1, 6.0 Hz, 3H),
3.64 (t, J=12.0 Hz, 3H), 3.58-3.49 (m, 18H), 3.47 (t, J=5.4 Hz,
6H), 3.37-3.32 (m, 6H), 3.17 (s, 9H), 3.06 (t, J=5.5 Hz, 6H),
2.26-2.16 (m, 6H), 2.05-1.96 (m, 6H). MS (ES, m/z): 1813.1
[M+H].sup.+.
Example 7
18-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenyl-
sulfonamido)-6,6-bis(3-(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetr-
ahydroisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethylamino)-3-oxopr-
opyl)-4,9-dioxo-13,16-dioxa-3,5,10-triazaoctadecan-1-oic acid
##STR00039##
[0374] Example 7: To a solution of example 2 (150 mg, 0.0882 mmol)
and DIEA (30 .mu.L, 0.18 mmol) in THF (3 mL) was added ethyl
2-isocyanatoacetate (20 .mu.L, 0.18 mmol) and the resulting
solution stirred for 1.5 h at room temperature. H.sub.2O (2 mL) and
LiOH.H.sub.2O (18.5 mg, 0.441 mmol) was then added and the
resulting mixture stirred for 2 h at room temperature. The mixture
was diluted with DCM and washed with H.sub.2O, then the organic
layer dried over Na.sub.2SO.sub.4 and the solvent remove under
reduced pressure. The resulting residue was then purified by
preparative HPLC with a C18 silica gel stationary phase using a
gradient of H.sub.2O 0.05% TFA:CH.sub.3CN 0.05% TFA (70:30 to
40:60) and detection by UV at 254 nm to give the title compound
tri-TFA salt (94 mg, 25% yield) as a white solid. .sup.1H-NMR (400
MHz, CD.sub.3OD) .delta. 7.92-7.86 (m, 3H), 7.79 (t, J=1.6 Hz, 3H),
7.65 (t, J=7.8 Hz, 3H), 7.58-7.52 (m, 6H), 6.84 (s, 3H), 4.80 (dd,
J=16.8, 5.4 Hz, 6H), 4.51 (d, J=16.1 Hz, 3H), 3.91 (dd, J=12.6, 6.4
Hz, 3H), 3.77 (s, 2H), 3.63 (t, J=12.0 Hz, 3H), 3.58-3.50 (m, 18H),
3.47 (t, J=5.4 Hz, 6H), 3.36-3.32 (m, 6H), 3.17 (s, 9H), 3.06 (t,
J=5.5 Hz, 6H), 2.25-2.15 (m, 6H), 1.96-1.85 (m, 6H). MS (ES, m/z):
1798.1 [M+H].sup.+.
Example 8
N1,N7-bis(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquino-
lin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethyl)-4-(3-(2-(2-(2-(3-((S)-6,8--
dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfonamido)eth-
oxy)ethoxy)ethylamino)-3-oxopropyl)-4-ureidoheptanediamide
##STR00040##
[0376] Example 8: To a solution of example 2 (50 mg, 0.029 mmol)
and TEA (12 .mu.L, 0.088 mmol) in DCM (1 mL) was added
isocyanatotrimethylsilane (5.9 .mu.L, 0.044 mmol). The resulting
solution was stirred for 1 h at room temperature, then stirred for
an additional 16 h at 40.degree. C. The solvent was then removed
under reduced pressure and the crude residue purified by
preparative HPLC with a C18 silica gel stationary phase using a
gradient of H.sub.2O 0.05% TFA:CH.sub.3CN 0.05% TFA (80:20 to
20:80) and detection by UV at 254 nm to give the title compound
tri-TFA salt (26 mg, 42% yield) as a white solid. .sup.1H-NMR (400
MHz, CD.sub.3OD) .delta. 7.89 (d, J=7.9 Hz, 3H), 7.79 (t, J=5.7 Hz,
3H), 7.65 (t, J=7.8 Hz, 3H), 7.58-7.50 (m, 6H), 6.84 (s, 3H), 4.80
(d, J=12.2 Hz, 6H), 4.51 (d, J=15.6 Hz, 3H), 3.92 (dd, J=12.2, 5.7
Hz, 3H), 3.64 (t, J=12.0 Hz, 3H), 3.58-3.49 (m, 18H), 3.47 (t,
J=5.4 Hz, 6H), 3.36-3.32 (m, 6H), 3.17 (d, J=5.5 Hz, 9H), 3.06 (t,
J=5.4 Hz, 6H), 2.25-2.15 (m, 5H), 2.03-1.86 (m, 6H). MS (ES, m/z):
1740.1 [M+H].sup.+.
Example 9
4-amino-4-(1-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-
-yl)phenylsulfonamido)-13-oxo-3,6,9-trioxa-12-azapentadecan-15-yl)-N.sup.1-
,N.sup.7-bis(2-(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydrois-
oquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethoxy)ethyl)heptanediamide
##STR00041##
[0378] Example 9: The title compound was synthesized in a manner
similar to example 2 (method B), using intermediate E in place of
intermediate A. MS (ES, m/z): 1829.2 [M+H].sup.+.
Example 10
4-(1-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phen-
ylsulfonamido)-13-oxo-3,6,9-trioxa-12-azapentadecan-15-yl)-N1,N7-bis(2-(2--
(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phe-
nylsulfonamido)ethoxy)ethoxy)ethoxy)ethyl)-4-propionamidoheptanediamide
##STR00042##
[0380] Example 10: Example 9 (75 mg, 0.041 mmol) and DIEA (21
.mu.L, 0.12 mmol) were dissolved in MeCN (1 mL) and cooled to
0.degree. C. To the stirring solution was added propionyl chloride
(4.3 .mu.L, 0.049 mmol), then the resulting mixture allowed to warm
to room temperature and stirred for 30 min. The mixture was diluted
with H.sub.2O and acidified with TFA, then purified by preparative
HPLC with a C18 silica gel stationary phase using a gradient of
H.sub.2O 0.05% TFA:CH.sub.3CN 0.05% TFA (80:20 to 20:80) and
detection by UV at 254 nm to give the title compound tri-TFA salt
(18 mg, 23% yield) as a white solid. .sup.1H-NMR (400 MHz,
CD.sub.3OD) .delta. 7.90 (d, J=7.9 Hz, 3H), 7.80-7.75 (m, 3H), 7.66
(t, J=7.8 Hz, 3H), 7.59-7.52 (m, 6H), 6.85 (s, 3H), 4.82-4.73 (m,
6H), 4.51 (d, J=16.4 Hz, 3H), 3.95-3.87 (m, 3H), 3.69-3.50 (m,
23H), 3.46 (t, J=5.4 Hz, 6H), 3.38-3.32 (m, 6H), 3.17 (s, 9H), 3.06
(t, J=5.4 Hz, 6H), 2.20-2.12 (m, 6H), 2.00-1.90 (m, 6H), 1.08 (t,
J=7.6 Hz, 3H). MS (ES, m/z): 1885.1 [M+H].sup.+.
Example 11
N.sup.1,N.sup.7-bis(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahyd-
roisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethyl)-4-(3-(2-(2-(2-(3-
-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfo-
namido)ethoxy)ethoxy)ethylamino)-3-oxopropyl)-4-(3-(1,3-dihydroxy-2-(hydro-
xymethyl)propan-2-yl)ureido)heptanediamide
##STR00043##
[0382] Example 11: To a solution of intermediate G (51 mg, 0.29
mmol) and DIEA (25 .mu.L, 0.15 mmol) dissolved in DMF (1 mL) was
added 2-Amino-2-(hydroxymethyl)-1,3-propanediol.HCl (9.2 mg, 0.059
mmol). The resulting solution was stirred at room temperature for 2
h, then diluted with H.sub.2O, and acidified with TFA. The mixture
was then purified by preparative HPLC with a C18 silica gel
stationary phase using a gradient of H.sub.2O 0.05% TFA:CH.sub.3CN
0.05% TFA (80:20 to 40:60) and detection by UV at 254 nm to give
the title compound tri-TFA salt (17 mg, 26% yield) as a white
solid. .sup.1H-NMR (400 MHz, CD.sub.3OD) .delta. 7.92-7.86 (m, 3H),
7.80-7.76 (m, 3H), 7.65 (t, J=7.8 Hz, 3H), 7.58-7.53 (m, 6H), 6.84
(s, 3H), 4.83-4.74 (m, 6H), 4.52 (d, J=16.1 Hz, 3H), 3.91 (dd,
J=11.5, 6.1 Hz, 3H), 3.70-3.61 (m, 9H), 3.61-3.50 (m, 18H), 3.47
(t, J=5.4 Hz, 6H), 3.34 (t, J=5.5 Hz, 6H), 3.17 (s, 9H), 3.06 (t,
J=5.4 Hz, 6H), 2.20 (dd, J=9.8, 6.4 Hz, 6H), 1.91 (dd, J=9.8, 6.5
Hz, 6H). MS (ES, m/z): 1844.0 [M+H].sup.+.
Example 12
(S)-2-(3-(1,25-bis(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquino-
lin-4-yl)phenylsulfonamido)-13-(3-(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1-
,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethylami-
no)-3-oxopropyl)-10,16-dioxo-3,6,20,23-tetraoxa-9,17-diazapentacosan-13-yl-
)ureido)succinic acid
##STR00044##
[0384] Example 12: To a solution of intermediate G (50 mg, 0.029
mmol) and DIEA (10 .mu.L, 0.059 mmol) in MeCN (1 mL) was added
(S)-di-tert-butyl 2-aminosuccinate (11 mg, 0.044 mmol). The
resulting solution was stirred for 18 h at 40.degree. C., then
diluted with H.sub.2O, acidified with TFA, and then purified by
preparative HPLC with a C18 silica gel stationary phase using a
gradient of H.sub.2O 0.05% TFA: CH.sub.3CN 0.05% TFA (80:20 to
20:80) and detection by UV at 254 nm. The fractions with pure
material were combined and the lyophilized. The resulting solid was
dissolved in TFA and left at room temperature for 30 min, then
solvent removed under a stream of N.sub.2. The resulting residue
was dissolved in 1:1 MeCN: H.sub.2O and lyophilized to give the
title compound tri-TFA salt (13.5 mg, 21% yield) as a white solid.
.sup.1H-NMR (400 MHz, CD.sub.3OD) .delta. 7.92-7.87 (m, 3H), 7.80
(t, J=1.6 Hz, 3H), 7.65 (t, J=7.8 Hz, 3H), 7.59-7.51 (m, 6H), 6.84
(s, 3H), 4.80 (d, J=11.4 Hz, 6H), 4.57-4.44 (m, 4H), 3.92 (dd,
J=12.5, 6.1 Hz, 3H), 3.65 (d, J=12.1 Hz, 3H), 3.58-3.49 (m, 18H),
3.47 (t, J=5.4 Hz, 6H), 3.36-3.33 (m, 6H), 3.18 (s, 9H), 3.06 (t,
J=5.6 Hz, 6H), 2.79 (ddd, J=21.5, 16.7, 5.5 Hz, 2H), 2.24-2.13 (m,
6H), 1.93-1.83 (m, 6H). MS (ES, m/z): 1856.1 [M+H].sup.+.
Example 13
4-(1-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phen-
ylsulfonamido)-13-oxo-3,6,9-trioxa-12-azapentadecan-15-yl)-N.sup.1,N.sup.7-
-bis(2-(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinoli-
n-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethoxy)ethyl)-4-(3-phenylureido)hep-
tanediamide
##STR00045##
[0386] Example 13: The title compound was synthesized in a manner
similar to example 11, using example 9 in place of example 2 and
phenylisocyanate in place of isocyanatotrimethylsilane. MS (ES,
m/z): 1948.2 [M+H].sup.+.
Example 14
N.sub.1,N.sub.7-bis(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahyd-
roisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethyl)-4-(3-(2-(2-(2-(3-
-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfo-
namido)ethoxy)ethoxy)ethylamino)-3-oxopropyl)-4-(2-(dimethylamino)acetamid-
o)heptanediamide
##STR00046##
[0388] Example 14: To a solution of example 2 (100 mg, 0.0588
mmol), N,N-dimethylglycine (9.0 mg, 0.088 mmol), and DIEA (50
.mu.L, 0.29 mmol) in DMF (2 mL) was added HATU (27 mg, 0.071 mmol).
The resulting mixture was stirred for 2 h at room temperature, then
diluted with H.sub.2O and acidified with TFA, then purified by
preparative HPLC with a C18 silica gel stationary phase using a
gradient of H.sub.2O 0.05% TFA:CH.sub.3CN 0.05% TFA (80:20 to
40:60) and detection by UV at 254 nm to give the title compound
tri-TFA salt (70 mg, 53% yield) as a white solid. .sup.1H-NMR (400
MHz, CD.sub.3OD) .delta. 7.89 (d, J=7.9 Hz, 3H), 7.78 (d, J=1.6 Hz,
3H), 7.65 (t, J=7.8 Hz, 3H), 7.58-7.52 (m, 6H), 6.83 (s, 3H),
4.83-4.73 (m, 6H), 4.51 (d, J=15.7 Hz, 3H), 3.95-3.87 (m, 5H), 3.63
(t, J=12.1 Hz, 3H), 3.59-3.50 (m, 18H), 3.47 (t, J=5.4 Hz, 6H),
3.35 (t, J=5.5 Hz, 6H), 3.17 (s, 9H), 3.05 (t, J=5.4 Hz, 6H), 2.94
(s, 6H), 2.30-2.15 (m, 6H), 2.07-1.94 (m, 6H). MS (ES, m/z): 1782.2
[M+H].sup.+.
Example 15
4-(1-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phen-
ylsulfonamido)-13-oxo-3,6,9-trioxa-12-azapentadecan-15-yl)-N.sup.1,N.sup.7-
-bis(2-(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinoli-
n-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethoxy)ethyl)-4-undecanamidoheptane-
diamide
##STR00047##
[0390] Example 15: The title compound was synthesized in a manner
similar to example 14, using example 9 in place of example 2 and
undecanoic acid in place N,N-dimethylglycine. MS (ES, m/z): 1997.2
[M+H].sup.+.
Example 16
4-(4'-chlorobiphenyl-4-ylcarboxamido)-4-(1-(3-((S)-6,8-dichloro-2-methyl-1-
,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfonamido)-13-oxo-3,6,9-trioxa-1-
2-azapentadecan-15-yl)-N.sup.1,N.sup.7-bis(2-(2-(2-(2-(3-((S)-6,8-dichloro-
-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfonamido)ethoxy)etho-
xy)ethoxy)ethyl)heptanediamide
##STR00048##
[0392] Example 16: The title compound was synthesized in a manner
similar to Example 14, using example 9 in place of example 2 and
4'-chlorobiphenyl-4-carboxylic acid in place of
N,N-dimethylglycine. MS (ES, m/z): 1022.4 [M+2H].sup.2+.
Example 17
N.sup.1,N.sup.7-bis(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahyd-
roisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethyl)-4-(3-(2-(2-(2-(3-
-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfo-
namido)ethoxy)ethoxy)ethylamino)-3-oxopropyl)-4-(3-(2-morpholinoethyl)urei-
do)heptanediamide
##STR00049##
[0394] Intermediate 17a: To a stirring solution of di-tert-butyl
4-(3-tert-butoxy-3-oxopropyl)-4-isocyanatoheptanedioate (150 mg,
0.340 mmol) in THF (2 mL) was added 2-morpholinoethanamine (47
.mu.L, 0.36 mmol). The solution was stirred at room temperature for
2 h and then the solvent removed under reduced pressure. The
resulting residue was dissolved in formic acid and stirred at room
temperature for 16 h and the solvent was then removed under reduced
pressure to give
4-(2-carboxyethyl)-4-(3-(2-morpholinoethyl)ureido)heptanedioic acid
as a white solid, which was used directly without further
purification.
[0395] Example 17: To a stirring solution of 17a (24 mg, 0.060
mmol), intermediate A (100 mg, 0.199 mmol), and DIEA (102 .mu.L) in
DMF (1 mL) was added HATU (82 mg, 0.22 mmol). The resulting mixture
was stirred for 2 h at room temperature, then diluted with H.sub.2O
and acidified with TFA and purified by preparative HPLC with a C18
silica gel stationary phase using a gradient of H.sub.2O 0.05%
TFA:CH.sub.3CN 0.05% TFA (80:20 to 40:60) and detection by UV at
254 nm to give the title compound tri-TFA salt (63 mg, 45% yield)
as a white solid. .sup.1H-NMR (400 MHz, CD.sub.3OD) .delta. 7.89
(m, 3H), 7.78 (t, J=1.5 Hz, 3H), 7.65 (t, J=7.8 Hz, 3H), 7.60-7.51
(m, 6H), 6.83 (s, 3H), 4.82-4.71 (m, 6H), 4.49 (d, J=15.9 Hz, 3H),
4.03 (s, 2H), 3.89 (dd, J=11.6, 6.3 Hz, 3H), 3.85-3.75 (m, 2H),
3.69-3.51 (m, 21H), 3.48 (t, J=5.4 Hz, 6H), 3.36 (t, J=5.4 Hz, 6H),
3.25 (d, J=4.6 Hz, 2H), 3.15 (s, 9H), 3.05 (t, J=5.4 Hz, 6H),
2.46-2.35 (m, 2H), 2.29-2.13 (m, 6H), 2.05-1.92 (m, 6H). MS (ES,
m/z): 1853.1 [M+H].sup.+.
Example 18
(S,S)--N,N',N''-(2,2',2''-(2,2',2''-(2,2',2''-(2,2',2''-(4,4',4''-nitrilot-
ris(methylene)tris(1H-1,2,3-triazole-4,1-diyl))tris(ethane-2,1-diyl))tris(-
oxy)tris(ethane-2,1-diyl))tris(oxy)tris(ethane-2,1-diyl))tris(oxy)tris(eth-
ane-2,1-diyl))tris(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquino-
lin-4-yl)benzenesulfonamide)
##STR00050##
[0397] Example 18: To a solution of intermediate E.1 (70 mg, 0.158
mmol) and triprop-2-ynylamine (6.3 mg, 0.0478 mmol) in DMF (1 mL)
was added CuI (1.4 mg, 0.0072 mmol) and the resulting mixture
stirred for 16 h at room temperature. The mixture was then diluted
with H.sub.2O and acidified with TFA, then purified by preparative
HPLC with a C18 silica gel stationary phase using a gradient of
H.sub.2O 0.05% TFA:CH.sub.3CN 0.05% TFA (80:20 to 20:80) and
detection by UV at 254 nm to give the title compound tri-TFA salt
(40 mg, 36% yield) as a white solid. .sup.1H-NMR (400 MHz,
CD.sub.3OD) .delta. 8.37 (s, 3H), 7.86 (d, J=8.3 Hz, 3H), 7.77 (s,
3H), 7.63 (t, J=7.8 Hz, 3H), 7.57-7.50 (m, 6H), 6.82 (s, 3H),
4.84-4.74 (m, 6H), 4.66 (t, J=4.9 Hz, 6H), 4.55-4.44 (m, 8H),
3.97-3.83 (m, 8H), 3.71-3.38 (m, 35H), 3.16 (s, 9H), 3.01 (t, J=5.4
Hz, 6H). MS (ES, m/z): 1845.2 [M+H].sup.+.
Example 19
4-((4'-chlorobiphenyl-4-yl)methylamino)-4-(1-(3-((S)-6,8-dichloro-2-methyl-
-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfonamido)-13-oxo-3,6,9-trioxa-
-12-azapentadecan-15-yl)-N.sup.1,N.sup.7-bis(2-(2-(2-(2-(3-((S)-6,8-dichlo-
ro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfonamido)ethoxy)et-
hoxy)ethoxy)ethyl)heptanediamide
##STR00051##
[0399] Example 19: A solution of example 9 (75 mg, 0.041 mmol) and
4'-chlorobiphenyl-4-carbaldehyde (8.9 mg, 0.041 mmol) in MeOH (1
mL) was stirred at room temperature for 4 h. NaBH.sub.4 (2.5 mg,
0.065 mmol) was then added and the resulting mixture stirred at
room temperature for 30 min. The solvent was then removed under
reduced pressure and the resulting residue dissolved in DCM and
washed with 1 M aqueous HCl, saturated aqueous NaHCO.sub.3, and
brine. The organic layer was then dried over Na.sub.2SO.sub.4 and
the solvent removed under reduced pressure. The residue was then
purified by preparative HPLC with a C18 silica gel stationary phase
using a gradient of H.sub.2O 0.05% TFA: CH.sub.3CN 0.05% TFA (80:20
to 20:80) and detection by UV at 254 nm to give the title compound
tri-TFA salt (26 mg, 25% yield) as a white solid. .sup.1H-NMR (400
MHz, CD.sub.3OD) .delta. 7.88 (d, J=7.9 Hz, 3H), 7.77 (s, 3H),
7.72-7.59 (m, 9H), 7.59-7.51 (m, 6H), 7.45 (m, 2H), 6.81 (s, 3H),
4.82-4.73 (m, 6H), 4.49 (d, J=16.2 Hz, 4H), 4.25 (s, 2H), 3.89 (dd,
J=12.2, 6.0 Hz, 3H), 3.68-3.48 (m, 33H), 3.45 (t, J=5.4 Hz, 6H),
3.40 (t, J=5.4 Hz, 6H), 3.15 (s, 9H), 3.05 (t, J=5.4 Hz, 6H), 2.50
(t, J=7.2 Hz, 6H), 2.13 (t, J=7.2 Hz, 6H). MS (ES, m/z): 1015.3
[M+2H].sup.2+.
Example 20
4-(1-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phen-
ylsulfonamido)-13-oxo-3,6,9-trioxa-12-azapentadecan-15-yl)-N.sup.1,N.sup.7-
-bis(2-(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinoli-
n-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethoxy)ethyl)-4-(4-(octyloxy)benzyl-
amino)heptanediamide
##STR00052##
[0401] Example 20: The title compound was synthesized in a manner
similar to example 19, 4-(octyloxy)benzaldehyde in place of
4'-chlorobiphenyl-4-carbaldehyde. MS (ES, m/z): 1024.5
[M+2H].sup.2+.
Examples 22-41
##STR00053##
[0403] The following examples were synthesized using methods
similar to the prior example given in Table 2 below:
TABLE-US-00002 TABLE 2 Example Exact Observed # R Method Mass Mass
22 ##STR00054## Example 14 1864.6 1865.3 [M + H].sup.+ 23
##STR00055## Example 19 1914.6 1915.2 [M + H].sup.+ 24 ##STR00056##
Example 19 1896.5 1897.0 [M + H].sup.+ 25 ##STR00057## Example 11
1767.5 1768.2 [M + H].sup.+ 26 ##STR00058## Example 11 1753.5
1754.1 [M + H].sup.+ 27 ##STR00059## Example 6 1825.5 1826.1 [M +
H].sup.+ 28 ##STR00060## Example 6 1850.5 1851.5 [M + H].sup.+ 29
##STR00061## Example 11 1917.5 1918.2 [M + H].sup.+ 30 ##STR00062##
Example 14 1804.5 1805.4 [M + H].sup.+ 31 ##STR00063## Example 11
1840.5 1841.0 [M + H].sup.+ 32 ##STR00064## Example 11 1833.4
1834.0 [M + H].sup.+ 33 ##STR00065## Example 11 1896.6 1897.2 [M +
H].sup.+ 34 ##STR00066## Example 11 1813.5 1814.2 [M + H].sup.+ 35
##STR00067## Example 14 1910.5 1911.0 [M + H].sup.+ 36 ##STR00068##
Example 6 1810.5 1811.2 [M + H].sup.+ 37 ##STR00069## Example 11
1903.5 1903.9 [M + H].sup.+ 38 ##STR00070## Example 14 1806.5
1807.0 [M + H].sup.+ 39 ##STR00071## Example 11 1827.5 914.9 [M +
2H].sup.2+ 40 ##STR00072## Example 6 1796.5 1797.1 [M + H].sup.+ 41
##STR00073## Example 17 1822.5 1823.2 [M + H].sup.+
Example 42
3,12-bis(14-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4--
yl)phenylsulfonamido)-2-oxo-6,9,12-trioxa-3-azatetradecyl)-N.sup.1,N.sup.1-
4-bis(2-(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinol-
in-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethoxy)ethyl)-6,9-dioxa-3,12-diaza-
tetradecane-1,14-diamide
##STR00074##
[0405] Example 42:
3,12-bis(14-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-
-yl)phenylsulfonamido)-2-oxo-6,9,12-trioxa-3-azatetradecyl)-N1,N14-bis(2-(-
2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)p-
henylsulfonamido)ethoxy)ethoxy)ethoxy)ethyl)-6,9-dioxa-3,12-diazatetradeca-
ne-1,14-diamide. To a mixture of ethylene
glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid (40 mL,
0.105 mmol, 1 equiv) in DMF (1.5 mL) at rt were added DIEA (147
.mu.L, 0.84 mmol, 8 equiv) and HBTU (160 mg, 0.42 mmol, 4 equiv).
The mixture was stirred at rt for 0.5 h to give an activated
tetraacid mixture. To a mixture of intermediate E (104.9 mg, 0.193
mol, 1.83 equiv) in DMF (0.2 mL) at rt was added the activated
tetracid mixture (626 .mu.L) in portions over 20 minutes. The
mixture was stirred at rt for 1 h and purified by prep HPLC to give
44.2 mg (29%) of the title compound TFA salt as a pale yellow
solid. MS (ES, m/z): 831 [M+3H].sup.3+. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 7.89 (d, J=7.8 Hz, 4H), 7.77 (s, 4H), 7.66 (t,
J=7.8 Hz, 4H), 7.59-7.53 (m, 8H), 6.83 (s, 4H), 4.82-4.73 (m, 8H),
4.49 (d, J=15.9 Hz, 4H), 3.96-3.85 (m, 12H), 3.82-3.75 (m, 4H),
3.68-3.50 (m, 52H), 3.45 (dt, J=14.2, 5.5 Hz, 16H), 3.15 (s, 12H),
3.05 (t, J=5.3 Hz, 8H).
Example 43
(S)--N,N'-(15-(1-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinol-
in-4-yl)phenylsulfonamido)-10-oxo-3,6-dioxa-9,11-diazatetradecan-14-yl)-10-
,20-dioxo-3,6,24,27-tetraoxa-9,11,15,19,21-pentaazanonacosane-1,29-diyl)bi-
s(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)benzenes-
ulfonamide)
##STR00075##
[0407] Example 43:
(S)--N,N'-(15-(1-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquino-
lin-4-yl)phenylsulfonamido)-10-oxo-3,6-dioxa-9,11-diazatetradecan-14-yl)-1-
0,20-dioxo-3,6,24,27-tetraoxa-9,11,15,19,21-pentaazanonacosane-1,29-diyl)b-
is(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)benzene-
sulfonamide). To a mixture of N,N'-carbonyldiimidazole (56.6 mg,
0.349 mmol, 3 equiv) in DMF (0.6 mL) was added dropwise a solution
of tris(3-aminopropyl)amine (21.9 mg, 0.116 mmol, 1 equiv) in DMF
(0.4 mL). The mixture was stirred at rt for 3 h and used in next
step without purification. To a mixture of intermediate A (83.6 mg,
0.167 mol, 1.44 equiv) in DMF (0.2 mL) at 50.degree. C. was added
the above tris(3-aminopropyl)amine reaction mixture (560 .mu.L) in
portions over 30 minutes. The mixture was stirred at 50.degree. C.
for 2 h and purified by prep. HPLC to give 50.5 mg (41%) of the
title compound TFA salt as a white solid. MS (ES, m/z): 1770
[M+H].sup.+. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.89 (d,
J=7.8 Hz, 3H), 7.78 (s, 3H), 7.65 (t, J=7.8 Hz, 3H), 7.59-7.50 (m,
6H), 6.83 (s, 3H), 4.85-4.74 (m, 6H), 4.50 (d, J=16.0 Hz, 3H), 3.90
(dd, J=12.0, 6.3 Hz, 3H), 3.62 (t, J=12.0 Hz, 3H), 3.59-3.55 (m,
6H), 3.55-3.53 (m, 6H), 3.49 (dt, J=12.5, 5.4 Hz, 12H), 3.30-3.27
(m, 6H), 3.23 (t, J=6.3 Hz, 6H), 3.20-3.12 (m, 15H), 3.05 (t, J=5.4
Hz, 6H), 2.00-1.76 (m, 6H).
Example 44
[0408]
N.sup.1,N.sup.1,N.sup.12,N.sup.12-tetrakis(13-(3-((S)-6,8-dichloro--
2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfonamido)-4-oxo-8,11--
dioxa-3,5-diazatridecyl)dodecanediamide
##STR00076##
[0409] Intermediate 44a: dibenzyl
(((tert-butoxycarbonyl)azanediyl)bis(ethane-2,1-diyl))dicarbamate.
To a mixture of N,N'-di-2-diethylenetriamine (740 mg, 1.99 mmol, 1
equiv) in DCM (3 mL) at 0.degree. C. was added di-tert-butyl
dicarbonate (522 mg, 2.39 mol, 1.2 equiv) and TEA (0.416 mL, 2.99
mmol, 1.5 equiv). The mixture was stirred at rt overnight, diluted
with ethyl acetate, washed with H.sub.2O (1.times.) and brine
(1.times.), dried, concentrated and purified by column to give
0.918 mg (98%) of intermediate 44a as clear syrup.
[0410] Intermediate 44b: tert-butyl bis(2-aminoethyl)carbamate. To
a mixture of dibenzyl
(((tert-butoxycarbonyl)azanediyl)bis(ethane-2,1-diyl))dicarbamate
(918 mg, 1.95 mmol, 1.0 equiv) in MeOH (20 mL) was added 10% Pd/C
(150 mg). The mixture was stirred at rt under H.sub.2 for 1.5 h,
filtered and concentrated to give 400 mg (crude) of intermediate
44b as a white solid.
[0411] Intermediate 44c: tert-butyl
bis(13-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)p-
henylsulfonamido)-4-oxo-8,11-dioxa-3,5-diazatridecyl)carbamate. To
a mixture of N,N'-carbonyldiimidazole (88.8 mg, 0.548 mmol, 2
equiv) in DMF (1.4 mL) was added dropwise a mixture of tert-butyl
bis(2-aminoethyl)carbamate (55.6 mg, 0.274 mmol, 1 equiv). The
mixture was stirred at rt for 3 h and used in next step without
purification. To a mixture of intermediate A (191.5 mg, 0.382 mol,
1.39 equiv) in DMF (0.4 mL) at 50.degree. C. was added portionwise
the above tert-butyl bis(2-aminoethyl)carbamate reaction mixture
and stirred at 50.degree. C. for 1 h. The mixture was diluted with
ethyl acetate, washed with H.sub.2O (2.times.) and brine
(1.times.), dried, concentrated and purified by column to give 172
mg (71%) of intermediate 44c as clear syrup.
[0412] Intermediate 44d:
(S)--N,N'-(10,18-dioxo-3,6,22,25-tetraoxa-9,11,14,17,19-pentaazaheptacosa-
ne-1,27-diyl)bis(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinoli-
n-4-yl)benzenesulfonamide). To a mixture of tert-butyl
bis(13-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)p-
henylsulfonamido)-4-oxo-8,11-dioxa-3,5-diazatridecyl)carbamate (172
mg) in DCM (0.5 mL) was added a solution of HCl in dioxane (4 M, 2
mL). The mixture was stirred at rt for 30 minutes and concentrated
to give 200 mg of intermediate 44d HCl salt as a white solid.
[0413] Example 44:
N.sup.1,N.sup.1,N.sup.12,N.sup.12-tetrakis(13-(3-((S)-6,8-dichloro-2-meth-
yl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfonamido)-4-oxo-8,11-dioxa--
3,5-diazatridecyl)dodecanediamide. To a mixture of
(S)--N,N'-(10,18-dioxo-3,6,22,25-tetraoxa-9,11,14,17,19-pentaazaheptacosa-
ne-1,27-diyl)bis(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinoli-
n-4-yl)benzenesulfonamide) (44.3 mg, 0.0349 mmol, 1 equiv) and
dodecanedioic acid (3.66 mg, 0.0159 mmol, 0.46 equiv) in DMF (0.4
mL) at rt were added DIEA (30.4 .mu.L, 0.175 mmol, 5 equiv) and
HATU (14.6 mg, 0.0384 mmol, 1.1 equiv). The mixture was stirred at
rt for 0.5 h and purified by prep HPLC to give 28 mg (59%) of the
title compound TFA salt as a white solid. MS (ES, m/z): 1255
[M+2H].sup.2+. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.89 (d,
J=7.9 Hz, 4H), 7.79 (d, J=1.4 Hz, 4H), 7.65 (t, J=7.8 Hz, 4H),
7.59-7.51 (m, 8H), 6.83 (s, 4H), 4.83-4.73 (m, 8H), 4.52 (d, J=16.2
Hz, 4H), 3.92 (dd, J=12.2, 6.0 Hz, 4H), 3.64 (t, J=11.8 Hz, 4H),
3.58-3.49 (m, 16H), 3.49-3.36 (m, 24H), 3.26 (dd, J=13.0, 5.5 Hz,
16H), 3.17 (s, 12H), 3.05 (t, J=5.4 Hz, 8H), 2.37 (t, J=8.0 Hz,
4H), 1.61-1.48 (m, 4H), 1.36-1.21 (m, 12H).
Example 45
(S)--N,N'-(14-((1-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquino-
lin-4-yl)phenylsulfonamido)-14-(13-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-t-
etrahydroisoquinolin-4-yl)phenylsulfonamido)-4-oxo-8,11-dioxa-3,5-diazatri-
decyl)-10,15-dioxo-3,6-dioxa-9,11,14,16-tetraazaoctacosan-28-yl)carbamoyl)-
-10,18-dioxo-3,6,22,25-tetraoxa-9,11,14,17,19-pentaazaheptacosane-1,27-diy-
l)bis(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)benz-
enesulfonamide)
##STR00077##
[0415] Example 45:
(S)--N,N'-(14-((1-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquin-
olin-4-yl)phenylsulfonamido)-14-(13-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4--
tetrahydroisoquinolin-4-yl)phenylsulfonamido)-4-oxo-8,11-dioxa-3,5-diazatr-
idecyl)-10,15-dioxo-3,6-dioxa-9,11,14,16-tetraazaoctacosan-28-yl)carbamoyl-
)-10,18-dioxo-3,6,22,25-tetraoxa-9,11,14,17,19-pentaazaheptacosane-1,27-di-
yl)bis(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)ben-
zenesulfonamide). To a mixture of intermediate 44d (42.3 mg, 0.0365
mmol, 2.1 equiv) in DCM (0.25 mL) was added portionwise a solution
of 1,2-diisocyanatododecane (4.7 .mu.L, 0.0174 mmol, 1 equiv) and
TEA (4.8 .mu.L, 0.0347 mmol, 2 equiv) in DCM (0.1 mL). The mixture
was stirred at rt for 20 minutes, concentrated, and purified by
prep HPLC to give 31.6 mg (60%) of example 45 TFA salt as a white
solid. MS (ES, m/z): 1284.6 [M+2H].sup.2+. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 7.92-7.84 (m, 4H), 7.79 (s, 4H), 7.64 (t, J=7.8
Hz, 4H), 7.58-7.48 (m, 8H), 6.83 (s, 4H), 4.84-4.75 (m, 8H), 4.51
(d, J=15.9 Hz, 4H), 3.92 (dd, J=11.8, 6.5 Hz, 4H), 3.63 (t, J=12.1
Hz, 4H), 3.58-3.53 (m, 8H), 3.53-3.51 (m, 8H), 3.51-3.43 (m, 16H),
3.30-3.25 (m, 16H), 3.21 (t, J=6.8 Hz, 8H), 3.17 (s, 12H),
3.15-3.09 (m, 4H), 3.05 (t, J=5.4 Hz, 8H), 1.56-1.44 (m, 4H),
1.35-1.22 (m, 16H).
Example 46
(S)--N,N'-(14-amino-14-(13-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydr-
oisoquinolin-4-yl)phenylsulfonamido)-4-oxo-8,11-dioxa-3,5-diazatridecyl)-1-
0,18-dioxo-3,6,22,25-tetraoxa-9,11,17,19-tetraazaheptacosane-1,27-diyl)bis-
(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)benzenesu-
lfonamide)
##STR00078##
[0417] Intermediate 46b: To a mixture of intermediate 46a (Ref 1,
411 mg, 0.893 mmol, 1 equiv) and DIEA (238.6 .mu.L, 1.34 mmol, 1.5
equiv) in DCM (3.3 mL) at 0.degree. C. was added Fmoc-OSu (362 mg,
1.07 mmol, 1.2 equiv). The mixture was stirred at rt for 4 h,
concentrated and purified by column to give 0.59 g (97%) of
intermediate 46b as a white solid.
[0418] Intermediate 46c: (9H-fluoren-9-yl)methyl
(1,5-diamino-3-(2-aminoethyl)pentan-3-yl)carbamate. To intermediate
46b was added a solution of HCl in dioxane (3 mL). The mixture was
stirred at rt for 0.5 h, concentrated and triturated with ethyl
acetate to give 0.216 g of intermediate 46c as a white solid.
[0419] Intermediate 46c1: (9H-fluoren-9-yl)methyl
(1,27-bis(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl-
)phenylsulfonamido)-14-(13-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydr-
oisoquinolin-4-yl)phenylsulfonamido)-4-oxo-8,11-dioxa-3,5-diazatridecyl)-1-
0,18-dioxo-3,6,22,25-tetraoxa-9,11,17,19-tetraazaheptacosan-14-yl)carbamat-
e. To a mixture of phosgene (15% in toluene, 577 .mu.L, 0.808 mmol,
2 equiv) in DCM (1 mL) at 0.degree. C. was added dropwise a mixture
of intermediate A (202.4 mg, 0.404 mmol, 1 equiv) and TEA (113
.mu.L, 0.808 mmol, 2 equiv) in DCM (3 mL). The mixture was stirred
at rt for 0.5 h and concentrated. The residue was diluted with THF
and filtered. The filtrate was concentrated to give a yellow solid.
To a mixture of this yellow solid in DMF (2.5 mL) was added
intermediate 46c (52.2 mg, 0.106 mmol, 0.263 equiv) and DIEA (141
.mu.L, 0.808 mmol, 2 equiv). The mixture was stirred at rt for 1 h
and diluted with water. The yellow precipitate was collected via
filtration and purified by column to give 122 mg (58%) of
intermediate 46d as a slightly yellow solid.
[0420] Example 46:
(S)--N,N'-(14-amino-14-(13-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahyd-
roisoquinolin-4-yl)phenylsulfonamido)-4-oxo-8,11-dioxa-3,5-diazatridecyl)--
10,18-dioxo-3,6,22,25-tetraoxa-9,11,17,19-tetraazaheptacosane-1,27-diyl)bi-
s(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)benzenes-
ulfonamide) To a mixture of intermediate 46d (122 mg, 0.062 mmol, 1
equiv) in DMF (2 mL) was added tris(2-aminoethyl)amine. The mixture
was stirred at rt for 0.5 h and purified by prep HPLC to give 81.5
mg (60%) of the title compound TFA salt as a white solid. MS (ES,
m/z): 1742.8 [M+H].sup.+. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
7.92-7.86 (m, 3H), 7.78 (t, J=1.6 Hz, 3H), 7.65 (t, J=7.8 Hz, 3H),
7.59-7.51 (m, 6H), 6.83 (s, 3H), 4.83-4.74 (m, 6H), 4.50 (d, J=16.0
Hz, 3H), 3.95-3.85 (m, 3H), 3.62 (t, J=12.1 Hz, 3H), 3.58-3.54 (m,
6H), 3.53 (dd, J=3.7, 1.7 Hz, 6H), 3.49 (dt, J=10.7, 5.5 Hz, 12H),
3.30-3.22 (m, 12H), 3.16 (s, 9H), 3.05 (t, J=5.4 Hz, 6H), 1.87 (t,
J=8.0 Hz, 6H).
Example 47
(S)--N,N'-(15-(1-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinol-
in-4-yl)phenylsulfonamido)-11-methyl-10-oxo-3,6-dioxa-9,11-diazatetradecan-
-14-yl)-11,19-dimethyl-10,20-dioxo-3,6,24,27-tetraoxa-9,11,15,19,21-pentaa-
zanonacosane-1,29-diyl)bis(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydro-
isoquinolin-4-yl)benzenesulfonamide)
##STR00079##
[0422] Example 47:
(S)--N,N'-(15-(1-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquino-
lin-4-yl)phenylsulfonamido)-11-methyl-10-oxo-3,6-dioxa-9,11-diazatetradeca-
n-14-yl)-11,19-dimethyl-10,20-dioxo-3,6,24,27-tetraoxa-9,11,15,19,21-penta-
azanonacosane-1,29-diyl)bis(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydr-
oisoquinolin-4-yl)benzenesulfonamide). To a mixture of phosgene
(15% in toluene, 127 .mu.L, 0.178 mmol, 1.5 equiv) in DCM (0.2 mL)
at 0.degree. C. was added dropwise a mixture of intermediate A
(59.3 mg, 0.118 mmol, 1 equiv) and TEA (25 uL, 0.178 mmol, 1.5
equiv) in DCM (1 mL). The mixture was stirred at rt for 0.5 h and
concentrated. To a mixture of the above isocyanate residue in DCM
(0.3 mL) was added a mixture of
N.sup.1-methyl-N.sup.3,N.sup.3-bis(3-(methylamino)propyl)propan-1,3-diami-
ne (intermediate J, 9 mg, 0.039 mmol, 0.33 equiv) and TEA (16.4
.mu.L, 0.118 mmol, 1 equiv) in DCM (0.44 mL) portionwise. The
mixture was stirred at rt for 0.5 h, concentrated and purified by
prep HPLC to give 51.3 mg (57%) of the title compound TFA salt as a
white solid. MS (ES, m/z): 1812.1 [M+H].sup.+. .sup.1H NMR (400
MHz, CD.sub.3OD) .delta. 7.92-7.86 (m, 3H), 7.78 (t, J=1.6 Hz, 3H),
7.65 (t, J=7.8 Hz, 3H), 7.59-7.51 (m, 6H), 6.82 (s, 3H), 4.84-4.73
(m, 6H), 4.50 (d, J=16.0 Hz, 3H), 3.96-3.85 (m, 3H), 3.63 (t,
J=12.1 Hz, 3H), 3.59-3.55 (m, 6H), 3.55-3.50 (m, 12H), 3.47 (t,
J=5.4 Hz, 6H), 3.40 (t, J=6.6 Hz, 6H), 3.35 (t, J=5.8 Hz, 6H), 3.16
(s, 9H), 3.15-3.10 (m, 6H), 3.05 (t, J=5.4 Hz, 6H), 2.92 (s, 9H),
2.05-1.86 (m, 6H).
Examples 48-71
##STR00080## ##STR00081## ##STR00082## ##STR00083## ##STR00084##
##STR00085## ##STR00086## ##STR00087## ##STR00088##
##STR00089##
TABLE-US-00003 [0423] TABLE 3 Example Method used Calc. MS Observed
MS Example 48 Example 42 2454.7 1228.6 [M + 2H].sup.2+ Example 49
Example 42 3028.8 1011 [M + 3H)].sup.3+ Example 50 Example 42
2312.6 1157 [M + 2H].sup.2+ Example 51 Example 43 1901.6 1903 [M +
H].sup.+ Example 52 Example 43 1859.6 1860.4 [M + H].sup.+ Example
53 Example 43 1727.5 1728 [M + H].sup.+ Example 54 Example 43
1811.6 1812.2 [M + H].sup.+ Example 55 Example 43 1811.6 1812.2 [M
+ H].sup.+ Example 56 Example 43 1901.6 1902.2 [M + H].sup.+
Example 57 Example 47 1853.6 1854.2 [M + H].sup.+ Example 58
Example 47 1784.5 1785.5 [M + H].sup.+ Example 59 Example 44 2616.8
1309.5 [M + 2H].sup.2+ Example 60 Example 45 2454.7 1228.5 [M +
2H].sup.2+ Example 61 Example 44 2792.9 1397.5 [M + 2H].sup.2+
Example 62 Example 46 1783.5 1784.1 [M + H].sup.+ Example 63
Example 6 1883.6 1884.2 [M + H].sup.+ Example 64 Example 2 1899.5
1900.2 [M + H].sup.+ Example 65 Example 11 1990.6 1992.1 [M +
H].sup.+ Example 66 Example 6 1857.5 1858.2 [M + H].sup.+ Example
67 Example 2 1738.5 1739.6 [M + H].sup.+ Example 68 Example 14
1823.4 1824.4 [M + H].sup.+ Example 69 Example 7 1839.5 1840.3 [M +
H].sup.+ Example 70 Example 6 1854.5 1855.4 [M + H].sup.+ Example
71 Example 4 1766.5 1767.2 [M + H].sup.+
Example 72
4-acetyl-N1,N7-bis(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydr-
oisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethyl)-4-(3-(2-(2-(2-(3--
((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfon-
amido)ethoxy)ethoxy)ethylamino)-3-oxopropyl)heptanediamide
##STR00090##
[0425] Example 72. HATU (42 mg, 0.11 mmol) was added to a solution
of 4-acetyl-4-(2-carboxyethyl)heptanedioic acid (intermediate N,
8.2 mg, 0.03 mmol),
(S)--N-(2-(2-(2-aminoethoxyl)ethoxy)ethyl)-3-(6,8-dichloro-2-methyl-1,2,3-
,4-tetrahydroisoquinolin-4-yl)benzenesulfonamide (intermediate A,
50 mg, 0.10 mmol) and DIEA (28 mg, 0.022 mmol) in DMF (0.50 mL).
After 30 minutes, the solvent was removed under vacuum and the
residue was purified by reverse phase HPLC (ACN/water/0.1% TFA) to
give a TFA salt of the title compound (13 mg). .sup.1H-NMR (400
MHz, CD.sub.3OD): .delta. 7.87 (ddd, J.sub.1,2=7.8 Hz,
J.sub.1,3=1.8 Hz, J.sub.1,4=1.2 Hz, 3H), 7.75 (t, J=1.8 Hz, 3H),
7.63 (t, J=7.7 Hz), 7.53 (m, 6H), 6.82 (s, 3H), 4.84-4.74 (m, 6H),
4.49 (d, J=16.2 Hz, 3H), 3.89, (dd, J.sub.1,2=6.1 Hz,
J.sub.1,3=12.3 Hz, 3H), 3.61 (t, J=12.1 Hz, 3H), 3.55-3.48 (m,
18H), 3.45 (t, J=5.5 Hz, 6H), 3.30 (m, 6H), 3.15 (s, 9H), 3.03 (t,
J=5.5 Hz, 6H), 2.14 (s, 3H), 2.05-2.01 (m, 6H), 1.85-1.81 (m, 6H).
MS (m/z): 1724.3 (M+H).
Example 73
N1,N7-bis(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquino-
lin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethyl)-4-(3-(2-(2-(2-(3-((S)-6,8--
dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfonamido)eth-
oxy)ethoxy)ethylamino)-3-oxopropyl)-4-(1-hydroxyethyl)heptanediamide
##STR00091##
[0427] Example 73: Sodium borohydride (1 mg, 0.03 mmol) was added
to a solution of a TFA salt of
4-acetyl-N1,N7-bis(2-(2-(2-(3-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahyd-
roisoquinolin-4-yl)phenylsulfonamido)ethoxy)ethoxy)ethyl)-4-(3-(2-(2-(2-(3-
-((S)-6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)phenylsulfo-
namido)ethoxy)ethoxy)ethylamino)-3-oxopropyl)heptanediamide
(example 72, 15 mg, 0.007 mmol) in MeOH 200 .mu.L and DCM (54L).
After 2 hours, the reaction was concentrated at reduce pressure and
purified by reverse phase HPLC (ACN/water/0.1% TFA) to give a TFA
salt of the title compound (6.2 mg). .sup.1H-NMR (400 MHz,
CD.sub.3OD) .delta. 7.87 (ddd, J.sub.1,2=7.8 Hz, J.sub.1,3=1.8 Hz,
J.sub.1,4=1.2 Hz, 3H), 7.75 (t, J=1.8 Hz, 3H), 7.63 (t, J=7.8 Hz,
3H), 7.53 m, 6H), 6.82 (s, 3H), 4.79-4.74 (m, 6H), 4.49 (d, J=16.2
Hz, 3H), 3.88 (dd, J.sub.1,2=11.6 Hz, J.sub.1,3=6.1 Hz, 3H),
3.64-3.58 (m, 4H), 3.55-3.47 (m, 18H), 3.45 (t, J=5.4 Hz), 3.31 (m,
6H), 3.14 (s, 9H), 3.03 (t, J=5.5 Hz, 6H), 2.27-2.17 (m, 6H),
1.63-1.53 (m, 6H), 1.14 (d, J=5.7 Hz, 3H). MS (m/z): 1726.1
(M+H).
Pharmacological Data
Example 74
Cell-Based Assay of NHE-3 Activity
[0428] Rat or human NHE-3-mediated Natdependent H.sup.+ antiport
was measured using a modification of the pH sensitive dye method
originally reported by Paradiso (Proc. Natl. Acad. Sci. USA. (1984)
81(23): 7436-7440). Opossum kidney (OK) cells were obtained from
the ATCC and propagated per their instructions. The rat NHE-3 gene
(GenBank M85300) or the human NHE-3 gene (GenBank
NM.sub.--004174.1) was introduced into OK cells via
electroporation, and cells were seeded into 96 well plates and
grown overnight. Medium was aspirated from the wells, cells were
washed twice with NaCl-HEPES buffer (100 mM NaCl, 50 mM HEPES, 10
mM glucose, 5 mM KCl, 2 mM CaCl.sub.2, 1 mM MgCl.sub.2, pH 7.4),
then incubated for 30 min at room temperature with NH.sub.4Cl-HEPES
buffer (20 mM NH.sub.4Cl, 80 mM NaCl, 50 mM HEPES, 5 mM KCl, 2 mM
CaCl.sub.2, 1 mM MgCl.sub.2, pH 7.4) containing 5 .mu.M
bis(acetoxymethyl)
3,3'-(3',6'-bis(acetoxymethoxy)-5-((acetoxymethoxy)carbonyl)-3-oxo-3H-spi-
ro[isobenzofuran-1,9'-xanthene]-2',7'-diyl)dipropanoate (BCECF-AM).
Cells were washed twice with Ammonium free, Natfree HEPES (100 mM
choline, 50 mM HEPES, 10 mM glucose, 5 mM KCl, 2 mM CaCl.sub.2, 1
mM MgCl.sub.2, pH 7.4) and incubated in the same buffer for 10
minutes at room temperature to lower intracellular pH.
NHE-3-mediated recovery of neutral intracellular pH was initiated
by addition of Na-HEPES buffer containing 0.4 .mu.M ethyl isopropyl
amiloride (EIPA, a selective antagonist of NHE-1 activity that does
not inhibit NHE-3) and 0-30 .mu.M test compound, and monitoring the
pH sensitive changes in BCECF fluorescence (.lamda..sub.ex 505 nm,
.lamda..sub.em 538 nm) normalized to the pH insensitive BCECF
fluorescence (.lamda..sub.ex 439 nm, .lamda..sub.em 538 nm).
Initial rates were plotted as the average 2 or more replicates, and
pIC.sub.50 values were estimated using GraphPad Prism.
TABLE-US-00004 TABLE 4 Data for examples in human NHE3 inhibition
assay Example # Human NHE3 pIC.sub.50 1 B 2 C 3 C 4 C 5 B 6 C 7 C 8
C 9 C 10 C 11 B 12 C 13 B 14 C 15 B 16 A 17 C 18 B 19 A 20 C 21 B
22 B 23 C 24 A 25 B 26 C 27 C 28 C 29 C 30 B 31 C 32 B 33 C 34 C 35
B 36 C 37 C 38 C 39 C 40 C 41 C 42 C 43 C 44 A 45 B 46 C 47 C 48 C
49 B 50 B 51 C 52 C 53 B 54 C 55 C 56 C 57 C 58 C 59 C 60 C 61 C 62
C 63 C 64 C 65 C 66 C 67 C 68 C 69 C 70 C 71 C 72 B 73 C A NHE3
pIC.sub.50 < 5 B NHE3 pIC.sub.50 5-7 C NHE3 pIC.sub.50 >
7
Example 75
Inhibition of Intestinal Sodium Absorption
[0429] Urinary sodium concentration and fecal form were measured to
assess the ability of selected example compounds to inhibit the
absorption of sodium from the intestinal lumen. Eight-week old
Sprague-Dawley rats were purchased from Charles River Laboratories
(Hollister, Calif.), were housed 2 per cage, and acclimated for at
least 3 days before study initiation. Animals were fed Harlan
Teklad Global 2018 rodent chow (Indianapolis, Ind.) and water ad
libitum throughout the study and maintained in a standard
light/dark cycle of 6 AM to 6 PM. On the day of the study, between
4 PM and 5 PM, a group of rats (n=6) were dosed via oral gavage
with test compound at the dosage indicated or vehicle (water) at a
volume of 10 mL/kg. After dose administration animals were placed
in individual metabolic cages where they were also fed the same
chow in meal form and watered ad libitum. At 16 h post-dose, the
urine samples were collected and fecal form was assessed by two
independent observations. Fecal forms were scored according to a
common scale associated with increasing fecal water to the wettest
observation in the cage's collection funnel (1, normal pellet; 2,
pellet adhering to sides of collection funnel due to moisture; 3,
loss of normal pellet shape; 4, complete loss of shape with a
blotting pattern; 5, liquid fecal streams evident). A rat's fecal
form score (FFS) was determined by averaging both observational
scores for all rats within a group (n=6). In every study the
vehicle group average was 1. These averages are reported in Table
5. For urine samples, the volumes were determined gravimetrically
and centrifuged at 3,600.times.g. The supernatants were diluted
100-fold in deionized Milli-Q water then filtered through a 0.2
.mu.m GHP Pall AcroPrep filter plate (Pall Life Sciences, Ann
Arbor, Mich.) prior to analysis by ion chromatography. Ten
microliters of each filtered extract was injected onto a Dionex
ICS-3000 ion chromatograph system (Dionex, Sunnyvale, Calif.).
Cations were separated by an isocratic method using 25 mM
methanesulfonic acid as the eluent on an IonPac CS12A 2 mm
i.d..times.250 mm, 8 .mu.m particle size cation exchange column
(Dionex). Sodium was quantified using standards prepared from a
cation standard mix containing Li.sup.+, Na.sup.+, NH.sub.4.sup.+,
K.sup.+, Mg.sup.2+, and Ca.sup.2+ (Dionex). The mean mass of sodium
urinated for every group in the 16 h period was determined with the
vehicle group usually urinating approximately 21 mg sodium. The
urine Na (uNa) for rats in the test groups were expressed as a
percentage of the vehicle mean and the means were compared to that
of the vehicle group by utilizing a one-way analysis of variance
coupled with a Dunnett's post hoc test. Means that were
significantly lower than the vehicle group as determined by
statistical analysis were denoted: *, P<0.05; **, P<0.01;
***, P<0.001.
TABLE-US-00005 TABLE 5 Rat urinary sodium and fecal form 16 h
post-dose of test compound at 3 mg/kg Example # uNa (% of vehicle)
FFS 2 A*** 3 3 A*** 3 4 A*** 2 5 B*** 2 6 A*** 3 7 A*** 3 8 A*** 2
9 A** 1 10 B 2 11 B 2 12 B* 3 14 A*** 2 17 A*** 2 21 B 1 25 A*** 2
26 A*** 2 27 B** 2 28 B 2 29 B 2 30 C 1 31 A 2 32 B 2 33 A** 2 34
A*** 3 36 A*** 3 37 A*** 3 38 A** 3 39 A** 2 40 A*** 3 41 A** 3 43
B*** 2 47 B* 1 53 B* 2 54 C 2 55 B** 2 56 B 2 57 B 1 58 B 2 62 B***
2 63 B 2 64 B 2 65 B* 2 66 A* 2 67 C 1 68 B 1 70 B** 2 72 B** 2 73
B 2
In Table 5, A indicates that urine sodium was <35% of the
percentage of the vehicle mean; B indicates that urine sodium was
35-75% of the percentage of the vehicle mean; C indicates that
urine sodium was >75% of the percentage of the vehicle mean.
Example 76
Plasma PK
[0430] Sprague-Dawley rats (n=3) were dosed with test compounds by
oral gavage. Blood was collected at 0.5, 1, 2 and 4 h via
retro-orbital bleeds and processed to plasma using K.sub.2EDTA as
an anticoagulant. Plasma samples were treated with acetonitrile
containing an internal standard and precipitated proteins were
removed by centrifugation. Supernatants were analyzed by LC-MS/MS
and compound concentrations were determined by interpolation from a
calibration curve prepared in plasma. Accurate recovery of quality
control samples were confirmed to accept each analytical run. Table
6 illustrates data from the pharmacokinetic profiling of selected
example compounds. From studies in which one or more rats had
samples with test compound levels below the quantitative limit,
C.sub.max and AUC (reported as the mean of n=3) may be reported as
"<X" to indicate an upper bound.
TABLE-US-00006 TABLE 6 Plasma pharmacokinetics for example
compounds Nominal AUC Dose LLOQ Cmax (ng .times. Example (mg/kg)
(ng/mL) (ng/mL) hr/mL) 2 30 2 <4.0 <12.0 4 30 2 <5.0
<13.0 7 30 2 <2.0 <8.0 52 30 5 <5.0 <19.0
Example 77
Fecal Recovery
[0431] Three male Sprague Dawley rats were administered 1 mg/kg
test compound by oral gavage. Feces were collected from study
animals from 0-48 or 0-72 hours after dosing, dried by
lyophilization, and homogenized. Replicate aliquots of 40-60 mg
each were subjected to extraction/protein precipitation with 7:1
acetonitrile:water and centrifuged. Supernatants were diluted 1:10
in 50:50 acetonitrile:water prior to analysis by LC-MS/MS. Compound
concentrations, determined by interpolation from a standard
calibration curve prepared in blank feces matrix, were converted to
the percentage of dosed material recovered by taking into account
the total collected fecal matter. The percent recovery for each rat
was reported as the mean of the calculations from replicate
samples. The overall percent recovery (Fecal Recovery [%]) was
reported as the mean percent recovery from three rats. Accurate
quality control sample recoveries were confirmed in each run, and
extraction efficiency was periodically verified. Table 7
illustrates fecal recovery data for selected example compounds.
TABLE-US-00007 TABLE 7 Fecal recovery example compounds Nominal
Fecal Dose Collection Recovery Example # (mg/kg) Time (h) (%) 2 1
48 87 3 1 72 110.2 6 1 48 51.5 7 1 48 83.2 11 1 48 64.1 14 1 72
75.3 17 1 72 96.9 36 1 48 68 37 1 48 90.5 40 1 48 99.1 43 1 48 53.8
46 1 48 75.8 55 1 48 64.6 62 1 48 81.5 65 1 48 112.2
Example 78
Cell-Based Assay of NHE-3 Activity (Persistent Inhibition)
[0432] The ability of compounds to inhibit Rat NHE-3-mediated Nat
dependent H.sup.+ antiport after application and washout was
measured using a modification of the pH sensitive dye method
described above in Example 74. Opossum kidney (OK) cells were
obtained from the ATCC and propagated per their instructions. The
rat NHE-3 gene was introduced into OK cells via electroporation,
and cells were seeded into 96 well plates and grown overnight.
Medium was aspirated from the wells, cells were washed twice with
NaCl-HEPES buffer (100 mM NaCl, 50 mM HEPES, 10 mM glucose, 5 mM
KCl, 2 mM CaCl.sub.2, 1 mM MgCl.sub.2, pH 7.4), then overlayed with
NaCl-HEPES buffer containing 0-30 .mu.M test compound. After a 60
min incubation, the test drug containing buffer was aspirated from
the cells, cells were washed twice with NaCl-HEPES buffer without
drug, then incubated for 30 min at room temperature with
NH.sub.4Cl-HEPES buffer (20 mM NH.sub.4Cl, 80 mM NaCl, 50 mM HEPES,
5 mM KCl, 2 mM CaCl.sub.2, 1 mM MgCl.sub.2, pH 7.4) containing 5
.mu.M BCECF-AM. Cells were washed twice with ammonium free, Natfree
HEPES (100 mM choline, 50 mM HEPES, 10 mM glucose, 5 mM KCl, 2 mM
CaCl.sub.2, 1 mM MgCl.sub.2, pH 7.4) and incubated in the same
buffer for 10 minutes at room temperature to lower intracellular
pH. NHE-3-mediated recovery of neutral intracellular pH was
initiated (40 min after compound washout) by addition of Na-HEPES
buffer containing 0.4 .mu.M ethyl isopropyl amiloride (EIPA, a
selective antagonist of NHE-1 activity that does not inhibit
NHE-3), and monitoring the pH sensitive changes in BCECF
fluorescence (.lamda..sub.ex 505 nm, .lamda..sub.em 538 nm)
normalized to the pH insensitive BCECF fluorescence (.lamda..sub.ex
439 nm, .lamda..sub.em 538 nm). Initial rates were plotted as the
average 2 or more replicates, and pIC.sub.50 values were estimated
using GraphPad Prism.
TABLE-US-00008 TABLE 8 Data for examples in rat NHE-3 prompt and
persistent inhibition assays Rat NHE3 Rat NHE3 pIC.sub.50
pIC.sub.50 Example # (prompt) (persistent) 1 B C 2 C C 3 C C 4 C C
5 B C 6 C C 7 C C 8 B C 9 C C 10 C C 11 A C 12 C C 13 B C 14 C C 15
A B 16 B B 17 C C 18 B A 19 A C 20 B C 21 B C 22 A B 23 B B 24 A B
25 B C 26 B C 27 C C 28 B C 29 C C 30 B C 31 C C 32 C C 33 C C 34 C
C 36 C C 37 C C 38 B C 39 C C 40 C C 41 B C 42 C C 43 B C 44 A B 45
B A 46 C C 47 B C 48 B C 49 B B 51 C C 52 B C 53 B C 54 B C 55 B C
56 C C 57 B C 58 C C 59 B C 61 B B 62 B C 63 C C 64 C C 65 B C 66 C
C 67 C C 68 C C 69 C C 70 C C 72 B C 73 C C A NHE3 pIC.sub.50 <
5 B NHE3 pIC.sub.50 5-7 C NHE3 pIC.sub.50 > 7
Example 79
Pharmacokinetic Evaluation in Bile
[0433] Bile duct cannulated (BDC) Sprague-Dawley rats were dosed
with test compounds by oral gavage and a single aliquot of bile was
collected via cannula over the 24 h following dosing. Bile samples
were treated with acetonitrile and precipitated proteins were
removed by centrifugation. Some compounds required liquid-liquid
extraction using MTBE. After centrifugation, samples were diluted
as appropriate in mobile phase and analyzed by LC-MS/MS. The
concentrations of compounds in bile were determined by
interpolation from a standard calibration curve prepared in rat
bile from untreated BDC rats. Accurate recovery of quality control
samples was confirmed to accept each analytical run. Table 9
illustrates data from the bile exposure of selected example
compounds. Concentration in bile is reported in nM and represents
the mean result from n=3 rats.
TABLE-US-00009 TABLE 9 Bile concentration for example compounds
Nominal Dose Concentration in Example (mg/kg) Bile (nM) 2 30 4 3 30
10 4 30 45 6 30 21 7 30 19 14 30 28 36 30 17 40 30 23 43 30 6 46 30
3 62 30 7
[0434] All of the U.S. patents, U.S. patent application
publications, U.S. patent applications, foreign patents, foreign
patent applications and non-patent publications referred to in this
specification are incorporated herein by reference, in their
entirety to the extent not inconsistent with the present
description.
[0435] From the foregoing it will be appreciated that, although
specific embodiments of the invention have been described herein
for purposes of illustration, various modifications may be made
without deviating from the spirit and scope of the invention.
Accordingly, the invention is not limited except as by the appended
claims.
* * * * *