U.S. patent application number 12/173286 was filed with the patent office on 2009-01-15 for polyether brevetoxin derivatives as a treatment for cystic fibrosis, mucociliary dysfunction, and pulmonary diseases.
This patent application is currently assigned to University of North Carolina at Wilmington. Invention is credited to William M. Abraham, Daniel G. Baden, Andrea J. Bourdelais.
Application Number | 20090018184 12/173286 |
Document ID | / |
Family ID | 34375531 |
Filed Date | 2009-01-15 |
United States Patent
Application |
20090018184 |
Kind Code |
A1 |
Baden; Daniel G. ; et
al. |
January 15, 2009 |
Polyether Brevetoxin Derivatives as a Treatment for Cystic
Fibrosis, Mucociliary Dysfunction, and Pulmonary Diseases
Abstract
Disclosed are compounds that are derivatives of brevetoxin, or
PbTx, pharmaceutical formulations comprising the compounds, and
methods of regulating mucus transport in a cell, treating
mucociliary dysfunction and diseases related to decreased mucus
transport, wherein the compounds are of the Formula (I), and
Formula (III): ##STR00001## wherein R, R.sub.1, R.sub.2, R.sub.3,
A, n, and Y are as defined herein for each compound.
Inventors: |
Baden; Daniel G.;
(Wilmington, NC) ; Abraham; William M.; (Miami,
FL) ; Bourdelais; Andrea J.; (Wilmington,
NC) |
Correspondence
Address: |
MCDONNELL BOEHNEN HULBERT & BERGHOFF LLP
300 S. WACKER DRIVE, 32ND FLOOR
CHICAGO
IL
60606
US
|
Assignee: |
University of North Carolina at
Wilmington
|
Family ID: |
34375531 |
Appl. No.: |
12/173286 |
Filed: |
July 15, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10945467 |
Sep 20, 2004 |
7399782 |
|
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12173286 |
|
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60504665 |
Sep 19, 2003 |
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Current U.S.
Class: |
514/450 ;
549/275; 549/354 |
Current CPC
Class: |
A61P 11/06 20180101;
C07D 493/22 20130101; A61P 11/00 20180101; A61P 43/00 20180101;
A61P 11/12 20180101 |
Class at
Publication: |
514/450 ;
549/354; 549/275 |
International
Class: |
C07D 493/22 20060101
C07D493/22; A61K 31/366 20060101 A61K031/366; A61K 31/352 20060101
A61K031/352; A61P 11/00 20060101 A61P011/00 |
Claims
1. A compound of the Formula (I): ##STR00055## wherein A is
##STR00056## R is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8
cycloalkylcarbonyl, C.sub.2-C.sub.6 alkenyl, C.sub.1-C.sub.6 alkyl
ester, C.sub.2-C.sub.6 alkenyl ester, amino, amido, aryl ester,
cycloalkyl ester, cycloalkenyl ester, purinyl, pyrimidinyl,
heterocyclyl, aryl, or heteroaryl; R.sub.1 is H or --(CO)CH.sub.3;
and R.sub.2 and R.sub.3 at each occurrence are independently
--CH.sub.2(CO)CH.sub.3, --CH.sub.2(CO)CH.sub.2CH.sub.3,
--CH.sub.2(CO)CH(CH.sub.3).sub.2,
--CH.sub.2(CO)CH.sub.2CH.sub.2CH.sub.3,
--CH.sub.2(CO)CH(CH.sub.3)CH.sub.2CH.sub.3, or
--CH.sub.2(CO)CH.sub.2CH(CH.sub.3).sub.2, or OR.sub.2 and OR.sub.3
can be taken together to form a six membered ring of the formula
(Ia) ##STR00057## wherein X is CH.sub.2, C(O), or CH(CH.sub.3);
wherein the bracketed-dashed bonds indicate attachment to backbone;
Y is CH.dbd.CH, C (O), CH(CH.sub.3), or CH.sub.2; n is 1 or 0; and
with the proviso that when OR.sub.2 and OR.sub.3 are taken together
to form a ring of the formula (Ia), X is C.dbd.O and the double
bond is present; A is ##STR00058## Y is CH.dbd.CH, and R.sub.1 is
H, and n is 1, then R is not: ##STR00059## or a pharmaceutically
acceptable salt, solvate, ester, amide, hydrate, complex, or
combination thereof.
2. The compound according to claim 1, wherein R is ##STR00060##
##STR00061## ##STR00062## ##STR00063##
3. The compound according to claim 1, wherein R.sub.2 and R.sub.3
are each independently ##STR00064##
4. The compound according to claim 1, wherein R.sub.2 and R.sub.3
are taken together to form a ring of formula (Ia).
5. The compound according to claim 4, wherein R.sub.2 and R.sub.3
together form: ##STR00065## wherein the bracketed-dashed bonds
indicate the point of attachment to the backbone.
6. The compound according to claim 1, wherein the compound is of
the Formula (II): ##STR00066## (II).
7. A compound according to claim 1, wherein the compound is
##STR00067## ##STR00068## ##STR00069## ##STR00070## wherein R is
C.sub.1-C.sub.6 alkanoyl, C.sub.2-C.sub.6 alkenoyl, aroyl,
cycloalkanoyl, cycloalkenoyl, purinoyl, or pyrimidinoyl; and
R.sub.2 is ##STR00071##
8-10. (canceled)
11. A compound of the Formula (III): ##STR00072## wherein R is H,
OH, halogen, C.sub.1-C.sub.6 lower alkyl, C.sub.1-C.sub.6 alkyl
ester, C.sub.2-C.sub.6 alkenyl ester, amino, amido, aldehyde, aryl
ester, cycloalkyl ester, cycloalkenyl ester, purinyl, pyrimidinyl,
heterocyclyl, aryl, or heteroaryl; Y is C.dbd.O, CH.dbd.CH,
CHCH.sub.3 or CH.sub.2; and n is 1 or 0; or a pharmaceutically
acceptable salt, solvate, hydrate, amide, ester, complex, or
combination thereof.
12. A compound according to claim 11, of the formula (IV):
##STR00073##
13-18. (canceled)
19. A pharmaceutical formulation comprising a compound according to
claim 1, or a pharmaceutically acceptable salt, ester, amide,
solvate, hydrate, complex, or combination thereof; and at least one
pharmaceutically acceptable carrier, excipient, solvent, adjuvant
or diluent.
20. (canceled)
21. A pharmaceutical formulation comprising a compound according to
claim 11, or pharmaceutically acceptable salt, ester, amide,
solvate, hydrate, complex, or combination thereof, and at least one
pharmaceutically acceptable carrier, excipient, solvent, adjuvant
or diluent.
22. (canceled)
23. A method of treating diseases associated with decreased mucus
transport in a subject comprising administering to a subject in
need of such treatment a therapeutically effective amount of a
compound according to claim 1, or a pharmaceutically acceptable
salt, solvate, hydrate, ester, amide, complex, or combination
thereof, where the compound according to claim 1 has the formula:
##STR00074## wherein A is ##STR00075## R is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.1-C.sub.6 alkyl ester,
C.sub.2-C.sub.6 alkenyl ester, amino, amido, aryl ester, cycloalkyl
ester, cycloalkenyl ester, purinyl, pyrimidinyl, heterocyclyl,
aryl, or heteroaryl; R.sub.1 is H or --(CO)CH.sub.3; and R.sub.2
and R.sub.3 at each occurrence are independently
--CH.sub.2(CO)CH.sub.3, CH.sub.2 (CO)CH.sub.2CH.sub.3, --CH.sub.2
(CO)CH(CH.sub.3).sub.2, --CH.sub.2 (CO)CH.sub.2CH.sub.2CH.sub.3,
--CH.sub.2 (CO)CH(CH.sub.3) CH.sub.2CH.sub.3, or --CH.sub.2
(CO)CH.sub.2CH(CH.sub.3).sub.2, or OR.sub.2 and OR.sub.3 can be
taken together to form a six membered ring of the formula (Ia)
##STR00076## wherein X is CH, C.dbd.O, or CH(CH.sub.3); wherein the
bracketed-dashed bonds indicate attachment to backbone; Y is
CH.dbd.CH, C.dbd.O, CHCH.sub.3, or CH.sub.2; n is 1 or 0.
24. A method of treating diseases associated with decreased mucus
transport in a subject comprising administering to a subject in
need of such treatment a therapeutically effective amount of a
compound according to claim 11, or a pharmaceutically acceptable
salt, solvate, hydrate, ester, amide, complex, or combination
thereof.
25-27. (canceled)
28. A method of treating a subject who has, or is at increased risk
of developing chronic airway obstruction, asthma, pulmonary
disease, pulmonary infection, or cystic fibrosis, comprising
administering to a subject in need of such treatment a
therapeutically effective amount of a compound according to claim
1, or a pharmaceutically acceptable salt, solvate, hydrate, ester,
amide, complex, or combination thereof, where the compound
according to claim 1 has the formula: ##STR00077## wherein A is
##STR00078## R is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.1-C.sub.6 alkyl ester, C.sub.2-C.sub.6 alkenyl ester, amino,
amido, aryl ester, cycloalkyl ester, cycloalkenyl ester, purinyl,
pyrimidinyl, heterocyclyl, aryl, or heteroaryl; R.sub.1 is H or
--(CO)CH.sub.3; and R.sub.2 and R.sub.3 at each occurrence are
independently --CH.sub.2(CO)CH.sub.3, --CH.sub.2
(CO)CH.sub.2CH.sub.3, --CH.sub.2 (CO)CH(CH.sub.3).sub.2, --CH.sub.2
(CO)CH.sub.2CH.sub.2CH.sub.3, --CH.sub.2
(CO)CH(CH.sub.3)CH.sub.2CH.sub.3, or --CH.sub.2
(CO)CH.sub.2CH(CH.sub.3).sub.2, or OR.sub.2 and OR.sub.3 can be
taken together to form a six membered ring of the formula (Ia)
##STR00079## wherein X is CH, C.dbd.O, or CH(CH.sub.3); wherein the
bracketed-dashed bonds indicate attachment to backbone; Y is
CH.dbd.CH, C.dbd.O, CHCH.sub.3, or CH.sub.2; n is 1 or 0.
29. A method of treating a subject who has, or is at increased risk
of developing chronic airway obstruction, asthma, pulmonary
disease, pulmonary infection, or cystic fibrosis, comprising
administering to a subject in need of such treatment a
therapeutically effective amount of a compound according to claim
11, or a pharmaceutically acceptable salt, solvate, ester, amide,
hydrate, complex, or combination thereof.
30-31. (canceled)
32. A method for regulating mucus transport in a cell, comprising
contacting a cell with an effective amount of a compound according
to claim 1, or a pharmaceutically acceptable salt, solvate,
hydrate, ester, amide, complex, or combination thereof, where the
compound according to claim 1 has the formula: ##STR00080## wherein
A is ##STR00081## R is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.1-C.sub.6 alkyl ester, C.sub.2-C.sub.6 alkenyl
ester, amino, amido, aryl ester, cycloalkyl ester, cycloalkenyl
ester, purinyl, pyrimidinyl, heterocyclyl, aryl, or heteroaryl;
R.sub.1 is H or --(CO)CH.sub.3; and R.sub.2 and R.sub.3 at each
occurrence are independently --CH.sub.2(CO)CH.sub.3, CH.sub.2
(CO)CH.sub.2CH.sub.3, --CH.sub.2(CO)CH(CH.sub.3).sub.2,
--CH.sub.2(CO)CH.sub.2CH.sub.2CH.sub.3,
--CH.sub.2(CO)CH(CH.sub.3)CH.sub.2CH.sub.3, or --CH.sub.2
(CO)CH.sub.2CH(CH.sub.3).sub.2, or OR.sub.2 and OR.sub.3 can be
taken together to form a six membered ring of the formula (Ia)
##STR00082## wherein X is CH, C.dbd.O, or CH(CH.sub.3); wherein the
bracketed-dashed bonds indicate attachment to backbone; Y is
CH.dbd.CH, C.dbd.O, CHCH.sub.3, or CH.sub.2; n is 1 or 0.
33. A method for regulating mucus transport in a cell, comprising
contacting a cell with an effective amount of a compound according
to claim 11, or pharmaceutically acceptable salt, solvate, ester,
amide, hydrate, complex, or combination thereof.
34. A method of treating diseases associated with mucociliary
dysfunction in a subject comprising administering to a subject in
need of such treatment a therapeutically effective amount of a
compound according to claim 1, or a pharmaceutically acceptable
salt, solvate, hydrate, ester, amide, complex, or combination
thereof, where the compound according to claim 1 has the formula:
##STR00083## wherein A is ##STR00084## R is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.1-C.sub.6 alkyl ester,
C.sub.2-C.sub.6 alkenyl ester, amino, amido, aryl ester, cycloalkyl
ester, cycloalkenyl ester, purinyl, pyrimidinyl, heterocyclyl,
aryl, or heteroaryl; R.sub.1 is H or --(CO)CH.sub.3; and R.sub.2
and R.sub.3 at each occurrence are independently
--CH.sub.2(CO)CH.sub.3, --CH.sub.2 (CO)CH.sub.2CH.sub.3, --CH.sub.2
(CO)CH(CH.sub.3).sub.2, --CH.sub.2 (CO)CH.sub.2CH.sub.2CH.sub.3,
--CH.sub.2 (CO)CH(CH.sub.3)CH.sub.2CH.sub.3, or --CH.sub.2
(CO)CH.sub.2CH(CH.sub.3).sub.2, or OR.sub.2 and OR.sub.3 can be
taken together to form a six membered ring of the formula (Ia)
##STR00085## wherein X is CH, C.dbd.O, or CH(CH.sub.3); wherein the
bracketed-dashed bonds indicate attachment to backbone; Y is
CH.dbd.CH, C(O), CH(CH.sub.3), or CH.sub.2; n is 1 or 0.
35. A method of treating diseases associated with mucociliary
dysfunction in a subject comprising administering to a subject in
need of such treatment a therapeutically effective amount of a
compound according to claim 11, or pharmaceutically acceptable
salt, solvate, ester, amide, hydrate, complex, or combination
thereof.
36. A compound of the formula: ##STR00086## wherein A is
##STR00087## R is aryl, or heteroaryl, each of which is substituted
with up to five groups that are independently C.sub.1-C.sub.10
alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8
cycloalkyl(C.sub.1-C.sub.6)alkyl, C.sub.3-C.sub.8
cycloalkyl(C.sub.1-C.sub.6)alkoxy, C.sub.1-C.sub.10 alkoxy,
halogen, hydroxy, cyano, nitro, amino,
mono(C.sub.1-C.sub.6)alkylamino, di(C.sub.1-C.sub.6)alkylamino,
C.sub.2-C.sub.10alkenyl, C.sub.2-C.sub.10alkynyl, C.sub.1-C.sub.6
haloalkyl, C.sub.1-C.sub.6 haloalkoxy, amino(C.sub.1-C.sub.6)alkyl,
mono(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl or
di(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl; Y is
CH.dbd.CH, C(O), CH(CH.sub.3), or CH.sub.2; n is 1 or 0; and or a
pharmaceutically acceptable salt, solvate, amide, ester, hydrate,
complex, or combination thereof.
37-42. (canceled)
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional
Application 60/504,665, filed Sep. 19, 2003.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The invention relates to brevetoxin derivative compounds,
pharmaceutical formulations comprising the brevetoxin derivatives,
and methods of treating diseases that are related to decreased
mucus transport using the compounds and pharmaceutical
formulations.
[0004] 2. Description of the Related Art
[0005] Decreased mucus transport is characteristic of conditions
and diseases such as airway obstruction, asthma, increased
incidence of pulmonary disease and/or infection, and cystic
fibrosis. In particular, cystic fibrosis is characterized by
abnormal functioning of the airway epithelial cells. Cystic
fibrosis (or "CF") is caused by a defective gene that codes for a
Na.sup.+/Cl.sup.- transporter present on the surface of the
epithelial cells that line the trachea, lungs, and other organs,
including the intestines, pancreas, reproductive organs, and
kidneys. Hundreds of mutations have been identified in this gene,
all of which result in defective transport of sodium and chloride
by epithelial cells. The severity of the disease symptoms is
related to the inherited gene mutation or mutations. These
observations indicate that activation of sodium channels can lead
to bronchoconstriction and, in some cases, defects in mucus
transport, both of which are associated with airway diseases,
including CF.
[0006] The class of compounds known as the brevetoxins were
initially discovered when they were purified as toxins from
cultures of the Florida red tide organism Karenia brevis also known
as Gymnodinium breve and Ptychodiscus brevis (Baden, D. G., et al.,
Toxicon, 1982; 20(5):929-932). K. Brevis proliferates during red
tide incidents. The brevetoxins, also known as "PbTxw" toxins
(Ptychodiscus brevis toxin), have since been characterized and
found to be polycyclic-polyethers that initially were shown to have
binding activity to a unique site associated with rat brain
synaptosomes (Poli, M. A., et al., Molec. Pharm., 1986;
30:129-135). Brevetoxins are classified as neurotoxins that are
known to bind to voltage gated sodium channels. In particular, the
effects of brevetoxins are mediated by interaction with receptor
site 5 on the sodium channels. The general brevetoxin A and
brevetoxin B backbone structure are as follows, with PbTx molecules
(1-10) described.
##STR00002##
[0007] Generally, the activity of brevetoxins is thought to derive
from the general backbone structure. Ring A and intact rings H, I,
J, and K have been reported to be essential for the toxic activity
of these compounds. There have been no reports that link toxic
activity of brevetoxins to the various side chains appended to the
backbone structure.
[0008] .beta.-Naphthoyl-PbTx-3 is a brevetoxin derivative that
reduces sodium channel openings and effectively antagonizes the
actions of the native toxin in channel activation
(Purkerson-Parker, Chemistry and Biology, 2000; 7(6):385-393).
.beta.-Naphthoyl-PbTx-3 is thought to displace the native toxin
from its binding site, does not elicit opening of sodium channels
in the steady state, and findings indicate that it blocks
brevetoxin-induced opening of sodium channels.
[0009] If activation of voltage gated sodium channels causes airway
related diseases or conditions, effective modulation or blockade of
voltage gated sodium channels can be useful in alleviating airway
pathologies associated with mucociliary dysfunction, such as
asthma, chronic obstructive pulmonary diseases, pulmonary infection
(e.g., pneumonia, Pseudomonas), and cystic fibrosis. Thus, there is
a need for active agents that can act at the CFTR, P2Y.sub.2
receptors, A.sub.2B receptors, purinergic receptors, and chloride
ion channels, binding to voltage gated sodium channels, which are
useful in the regulation of mucus transport, as well as treatment
or prevention of conditions or diseases associated with decreased
mucus transport.
SUMMARY OF THE INVENTION
[0010] The invention provides compounds, or pharmaceutically
acceptable salts, solvates, hydrates, complexes, or combinations
thereof, of Formula (I):
##STR00003##
wherein [0011] A is
[0011] ##STR00004## [0012] R is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.3-C.sub.8 cycloalkylcarbonyl,
C.sub.1-C.sub.6 alkyl ester, C.sub.2-C.sub.6 alkenyl ester, amino,
amido, aryl ester, cycloalkyl ester, cycloalkenyl ester, purinyl,
pyrimidinyl, heterocyclyl, aryl, or heteroaryl, each of which is
optionally substituted on any available carbon atom with
C.sub.1-C.sub.10 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8
cycloalkyl(C.sub.1-C.sub.6)alkyl, C.sub.3-C.sub.8
cycloalkyl(C.sub.1-C.sub.6)alkoxy, C.sub.1-C.sub.10 alkoxy,
halogen, hydroxy, cyano, nitro, amino,
mono(C.sub.1-C.sub.6)alkylamino, di(C.sub.1-C.sub.6)alkylamino,
C.sub.2-C.sub.10alkenyl, C.sub.2-C.sub.10alkynyl, C.sub.1-C.sub.6
haloalkyl, C.sub.1-C.sub.6 haloalkoxy, amino(C.sub.1-C.sub.6)alkyl,
mono(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl or
di(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl; [0013] R.sub.1
is H or --(CO)CH.sub.3; and [0014] R.sub.2 and R.sub.3 at each
occurrence are independently --CH.sub.2(CO)CH.sub.3, --CH.sub.2
(CO)CH.sub.2CH.sub.3, [0015] --CH.sub.2 (CO)CH(CH.sub.3).sub.2,
--CH.sub.2 (CO)CH.sub.2CH.sub.2CH.sub.3, --CH.sub.2
(CO)CH(CH.sub.3)CH.sub.2CH.sub.3, or
--CH.sub.2(CO)CH.sub.2CH(CH.sub.3).sub.2, or OR.sub.2 and OR.sub.3
can be taken together to form a six membered ring of the formula
(Ia)
[0015] ##STR00005## [0016] wherein X is C.dbd.O, CH.sub.2, or
CH(CH.sub.3); [0017] wherein the bracketed-dashed bonds indicate
attachment to backbone; [0018] Y is CH.dbd.CH, C(O), CH(CH.sub.3),
or CH.sub.2; and [0019] n is 1 or 0; and [0020] with the proviso
that when OR.sub.2 and OR.sub.3 are taken together to form a ring
of the formula (Ia), wherein X is C.dbd.O and the double bond is
present; when A is
##STR00006##
[0020] and when n is 1, R is not:
##STR00007##
[0021] The invention also provides compounds, or pharmaceutically
acceptable salts, solvates, hydrates, complexes, or combinations
thereof, of the Formula (II):
##STR00008##
wherein [0022] A is
[0022] ##STR00009## [0023] R is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.1-C.sub.6 alkyl ester,
C.sub.2-C.sub.6 alkenyl ester, amino, amido, aryl ester, cycloalkyl
ester, cycloalkenyl ester, purinyl, pyrimidinyl, heterocyclyl,
aryl, or heteroaryl; [0024] R.sub.1 is H or --COCH.sub.3; and
[0025] R.sub.2 and R.sub.3 at each occurrence are independently
--CH.sub.2COCH.sub.3, --CH.sub.2COCH.sub.2CH.sub.3, [0026]
--CH.sub.2COCH(CH.sub.3).sub.2,
--CH.sub.2COCH.sub.2CH.sub.2CH.sub.3, --CH.sub.2COCH(CH.sub.3)
CH.sub.2CH.sub.3, or --CH.sub.2COCH.sub.2CH(CH.sub.3).sub.2, or
OR.sub.2 and OR.sub.3 can be taken together to form a six membered
ring of the formula (Ia)
[0026] ##STR00010## [0027] wherein X is C.dbd.O or CH(CH.sub.3);
[0028] wherein the bracketed-dashed bonds indicate attachment to
backbone; [0029] Y is CH.dbd.CH, C.dbd.O, or CH.sub.2; and [0030] n
is 1 or 0; and [0031] with the proviso that when OR.sub.2 and
OR.sub.3 are taken together to form a ring of the formula (Ia),
wherein X is C.dbd.O and the double bond is present; when A is
##STR00011##
[0031] and when n is 1, R is not:
##STR00012##
[0032] Further, the invention provides compounds, or
pharmaceutically acceptable salts, solvates, hydrates, complexes,
or combinations thereof, of the Formula (III):
##STR00013##
wherein [0033] R is H, OH, halogen, C.sub.1-C.sub.6 lower alkyl,
C.sub.1-C.sub.6 alkyl esters, C.sub.2-C.sub.6 alkenyl ester, amino,
amido, aldehydro such as formyl, aryl ester, cycloalkyl ester,
cycloalkenyl ester, purinyl, pyrimidinyl, heterocyclyl, aryl, or
heteroaryl; [0034] n is 1 or 0; and [0035] Y is C.dbd.O, CH.dbd.CH,
CHCH.sub.3 or CH.sub.2. [0036] Yet further, the instant invention
provides compounds, or pharmaceutically acceptable salts, solvates,
hydrates, complexes, or combinations thereof, of the Formula
(IV):
[0036] ##STR00014## [0037] wherein [0038] R is H, OH, halogen,
C.sub.1-C.sub.6 lower alkyl, C.sub.1-C.sub.6 alkyl ester,
C.sub.2-C.sub.6 alkenyl ester, amino, amido, aldehyde, aryl ester,
cycloalkyl ester, cycloalkenyl ester, purinyl, pyrimidinyl,
heterocyclyl, aryl, or heteroaryl; [0039] n is 1 or 0; and [0040] Y
is CH.dbd.CH, C.dbd.O, CH(CH.sub.3), or CH.sub.2.
[0041] The invention also provides pharmaceutical formulations
comprising a compound, or a pharmaceutically acceptable salt,
solvate, hydrate, complex, or combinations thereof, of Formulas
(I), (II), (III), and (IV) in combination with a pharmaceutically
acceptable carrier, excipient, solvent, adjuvant or diluent.
[0042] The invention further relates to methods of regulating mucus
transport velocity and treating conditions or diseases associated
with decreased mucus transport or mucociliary dysfunction in a
subject comprising administering to a subject a compound of Formula
(I), (II), (III), or (IV), or pharmaceutically acceptable salt,
solvate, hydrate, complex, or combination thereof.
BRIEF DESCRIPTION OF THE DRAWINGS
[0043] FIG. 1 illustrates the effects of PbTx-2 and PbTx-3 on
tracheal mucus velocity (TMV) in conscious allergic sheep, relative
to control (vehicle). PbTx-2 was relatively inactive, but PbTx-3
caused a rapid and pronounced decrease in TMV which was sustained
for up to 2 h. Values are mean.+-.sem.
[0044] FIG. 2 illustrates the effect of the .beta.-naphthoyl-PbTx-3
derivative on tracheal mucus velocity (TMV) in conscious sheep at
10 pg and 100 pg doses. The effect of the .beta.-naphthoyl
derivative at 10 pg (n=2) was about the same as control (vehicle).
At 100 pg, (n=4) the .beta.-naphthoyl derivative is able to rapidly
increase TMV that is sustainable for about 1 h. Values are
mean.+-.se.
[0045] FIG. 3 illustrates the differential effects of PbTx-3 (20
breaths of 10 pg/mL, n=4) and 10 pg (n=2) and 100 pg (n=4) doses of
the .beta.-naphthoyl-PbTx-3 antagonist on tracheal mucus velocity
in conscious sheep. Values are mean.+-.sem. Administration of the
.beta.-naphthoyl-PbTx-3 antagonist at each dosage increased TMV
relative to PbTx-3. Values are mean.+-.sem.
[0046] FIG. 4 illustrates the effect of the .beta.-naphthoyl-PbTx-3
derivative on PbTx-2-induced bronchoconstriction in conscious
sheep. Twenty breaths of increasing doses of PbTx-2 produced an
increase in pulmonary airflow resistance (RL). Pretreating the
animals with 20 breaths of 10 pg/mL of .beta.-naphthoyl-PbTx-3 at
15 minutes prior to PbTx-2 challenge blocked PbTx-2-induced
bronchoconstriction. Values are mean.+-.sem for 4-6 sheep.
[0047] FIG. 5 illustrates the effect of the .beta.-naphthoyl-PbTx-3
derivative on PbTx-3-induced bronchoconstriction in conscious
sheep. Twenty breaths of increasing doses of PbTx-3 produced an
increase in pulmonary airflow resistance (RL). Pretreating the
animals with 20 breaths of 10 pg/mL of .beta.-naphthoyl-PbTx-3 at
15 minutes prior to PbTx-3 challenge blocked PbTx-3-induced
constrictor response. Pretreatment of the animals with 20 breaths
of 100 pg/mL .beta.-naphthoyl-PbTx-3 at 15 minutes prior to PbTx-3
challenge, provided an increase the inhibition of PbTx-3-induced
bronchoconstriction. Values are mean.+-.sem for 4-6 sheep.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0048] Unless defined otherwise, all scientific and technical terms
used herein have the same meaning as commonly understood by one of
skill in the art to which this invention belongs.
[0049] All patents and publications referred to herein are hereby
incorporated by reference for all purposes.
[0050] A "therapeutically effective" amount is defined as an amount
effective to reduce or lessen at least one symptom of the disease
being treated or to reduce or delay onset of one or more clinical
markers or symptoms of the disease.
[0051] As used in this specification and the appended claims, the
singular forms "a," "an," and "the" include plural referents unless
the content clearly dictates otherwise. Thus, for example,
reference to a composition containing "a compound" includes a
mixture of two or more compounds. It should also be noted that the
term "or" is generally employed in its sense including "and/or"
unless the content clearly dictates otherwise.
[0052] By "alkyl" and "C.sub.1-C.sub.6 alkyl" is meant straight or
branched chain alkyl groups having 1-6 carbon atoms, such as,
methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl,
pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl,
and 3-methylpentyl. It is understood that in cases where an alkyl
chain of a substituent (e.g. of an alkyl, alkoxy or alkenyl group)
is shorter or longer than 6 carbons, it will be so indicated in the
second "C" as, for example, "C.sub.1-C.sub.10" indicates a maximum
of 10 carbons. The alkyl groups herein are optionally substituted
in one or more substitutable positions with various groups.
[0053] By the term "halogen" is meant fluorine, bromine, chlorine,
and iodine.
[0054] "Alkenyl" and "C.sub.2-C.sub.6 alkenyl" means straight and
branched hydrocarbon groups having from 2 to 6 carbon atoms and
from one to three double bonds and includes, for example, ethenyl,
propenyl, 1-but-3-enyl, 1-pent-3-enyl, 1-hex-5-enyl and the like.
The alkenyl groups herein are optionally substituted in one or more
substitutable positions with various groups.
[0055] As used herein, the term "cycloalkyl" refers to saturated
carbocyclic groups having three to twelve carbon atoms. The
cycloalkyl can be monocyclic, or a polycyclic fused system.
[0056] Examples of such groups include cyclopropyl, cyclobutyl,
cyclopentyl and cyclohexyl. The cycloalkyl groups herein are
unsubstituted or, as specified, substituted in one or more
substitutable positions with various groups. For example, such
cycloalkyl groups may be optionally substituted with, for example,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, halogen, hydroxy,
cyano, nitro, amino, mono(C.sub.1-C.sub.6)alkylamino,
di(C.sub.1-C.sub.6)alkylamino, C.sub.2-C.sub.6alkenyl,
C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
haloalkoxy, amino (C.sub.1-C.sub.6)alkyl,
mono(C.sub.1-C.sub.6)alkylamino (C.sub.1-C.sub.6)alkyl or
di(C.sub.1-C.sub.6)alkylamino (C.sub.1-C.sub.6)alkyl.
[0057] By "aryl" is meant an aromatic carbocyclic group having a
single ring (e.g., phenyl), multiple rings (e.g., biphenyl), or
multiple condensed rings in which at least one is aromatic, (e.g.,
1,2,3,4-tetrahydronaphthyl, naphthyl), which is optionally mono-,
di-, or trisubstituted. Preferred aryl groups of the invention are
phenyl, 1-naphthyl, 2-naphthyl, indanyl, indenyl, dihydronaphthyl,
tetralinyl or 6,7,8,9-tetrahydro-5H-benzo[a]cycloheptenyl. The aryl
groups herein are unsubstituted or, as specified, substituted in
one or more substitutable positions with various groups. Preferred
aryl groups are optionally substituted with C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkoxy, halogen, hydroxy, cyano, nitro, amino,
mono(C.sub.1-C.sub.6)alkylamino, di(C.sub.1-C.sub.6)alkylamino,
C.sub.2-C.sub.6alkenyl, C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6
haloalkyl, C.sub.1-C.sub.6 haloalkoxy, amino(C.sub.1-C.sub.6)alkyl,
mono(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl or
di(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl.
[0058] As used herein, the term "arylester" encompasses
aryloxycarbonyl and arylcarbonyloxy groups.
[0059] As used herein, the term "alkylester" encompasses
alkyloxycarbonyl and alkylcarbonyloxy groups. As used herein,
alkylcarbonyl carries the same meaning as alkanoyl.
[0060] As used herein, the term "alkylamide" encompasses
alkylaminocarbonyl groups, dialkylcarbonyl groups, and
alkanoylamino groups.
[0061] As used herein, the term "alkenylamide" encompasses
alkenylaminocarbonyl groups, dialkenylcarbonyl groups, and
alkenylcarbonylamino groups.
[0062] As used herein, the term "alkenylester" encompasses
alkenyloxycarbonyl and alkenylcarbonyloxy groups.
[0063] The term alkylarylester as used herein refers to
alkyloxycarbonyl and akanoyloxy groups in which the alkyl portion
carries an aryl or heteroaryl group.
[0064] The term alkenylarylester as used herein refers to
alkenyloxycarbonyl and alkenylcarbonyloxy groups in which the
alkenyl portion carries an aryl or heteroaryl group.
[0065] By "heteroaryl" is meant one or more aromatic ring systems
of 5-, 6-, or 7-membered rings which includes fused ring systems of
9-11 atoms containing at least one and up to four heteroatoms
selected from nitrogen, oxygen, or sulfur. Preferred heteroaryl
groups of the invention include pyridinyl, pyrimidinyl, quinolinyl,
benzothienyl, indolyl, indolinyl, pyridazinyl, pyrazinyl,
isoindolyl, isoquinolyl, quinazolinyl, quinoxalinyl, phthalazinyl,
imidazolyl, isoxazolyl, pyrazolyl, oxazolyl, thiazolyl,
indolizinyl, indazolyl, benzothiazolyl, benzimidazolyl,
benzofuranyl, furanyl, thienyl, pyrrolyl, oxadiazolyl,
thiadiazolyl, triazolyl, tetrazolyl, oxazolopyridinyl,
imidazopyridinyl, isothiazolyl, naphthyridinyl, cinnolinyl,
carbazolyl, beta-carbolinyl, isochromanyl, chromanyl,
tetrahydroisoquinolinyl, isoindolinyl, isobenzotetrahydrofuranyl,
isobenzotetrahydrothienyl, isobenzothienyl, benzoxazolyl,
pyridopyridinyl, benzotetrahydrofuranyl, benzotetrahydrothienyl,
purinyl, benzodioxolyl, triazinyl, phenoxazinyl, phenothiazinyl,
pteridinyl, benzothiazolyl, imidazopyridinyl, imidazothiazolyl,
dihydrobenzisoxazinyl, benzisoxazinyl, benzoxazinyl,
dihydrobenzisothiazyinyl, benzopyranyl, benzothiopyranyl,
coumarinyl, isocoumarinyl, chromonyl, chromanonyl,
pyridinyl-N-oxide, tetrahydroquinolinyl, dihydroquinolinyl,
dihydroquinolinonyl, dihydroisoquinolinonyl, dihydrocoumarinyl,
dihydroisocoumarinyl, isoindolinonyl, benzodioxanyl,
benzoxazolinonyl, pyrrolyl N-oxide, pyrimidinyl N-oxide,
pyridazinyl N-oxide, pyrazinyl N-oxide, quinolinyl N-oxide, indolyl
N-oxide, indolinyl N-oxide, isoquinolyl N-oxide, quinazolinyl
N-oxide, quinoxalinyl N-oxide, phthalazinyl N-oxide, imidazolyl
N-oxide, isoxazolyl N-oxide, oxazolyl N-oxide, thiazolyl N-oxide,
indolizinyl N-oxide, indazolyl N-oxide, benzothiazolyl N-oxide,
benzimidazolyl N-oxide, pyrrolyl N-oxide, oxadiazolyl N-oxide,
thiadiazolyl N-oxide, triazolyl N-oxide, tetrazolyl N-oxide,
benzothiopyranyl S-oxide, benzothiopyranyl S,S-dioxide. The
heteroaryl groups herein are unsubstituted or, as specified,
substituted in one or more substitutable positions with various
groups. Preferred heteroaryl groups are optionally substituted with
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, halogen, hydroxy,
cyano, nitro, amino, mono(C.sub.1-C.sub.6)alkylamino,
di(C.sub.1-C.sub.6)alkylamino, C.sub.2-C.sub.6alkenyl,
C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
haloalkoxy, amino(C.sub.1-C.sub.6)alkyl,
mono(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl or
di(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl.
[0066] By "heterocycle", "heterocycloalkyl" or "heterocyclyl" is
meant one or more carbocyclic ring systems of 4-, 5-, 6-, or
7-membered rings which includes fused ring systems of 9-11 atoms
containing at least one and up to four heteroatoms selected from
nitrogen, oxygen, or sulfur. Preferred heterocycles of the
invention include morpholinyl, thiomorpholinyl, thiomorpholinyl
S-oxide, thiomorpholinyl S,S-dioxide, piperazinyl, homopiperazinyl,
pyrrolidinyl, pyrrolinyl, tetrahydropyranyl, piperidinyl,
tetrahydrofuranyl, tetrahydrothienyl, homopiperidinyl,
homomorpholinyl, homothiomorpholinyl, homothiomorpholinyl
S,S-dioxide, oxazolidinonyl, dihydropyrazolyl, dihydropyrrolyl,
dihydropyrazinyl, dihydropyridinyl, dihydropyrimidinyl,
dihydrofuryl, dihydropyranyl, tetrahydrothienyl S-oxide,
tetrahydrothienyl S,S-dioxide and homothiomorpholinyl S-oxide. The
heterocycle groups herein are unsubstituted or, as specified,
substituted in one or more substitutable positions with various
groups. Preferred heterocycle groups are optionally substituted
with C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, halogen,
hydroxy, cyano, nitro, amino, mono(C.sub.1-C.sub.6)alkylamino,
di(C.sub.1-C.sub.6)alkylamino, C.sub.2-C.sub.6alkenyl,
C.sub.2-C.sub.6alkynyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
haloalkoxy, amino(C.sub.1-C.sub.6)alkyl,
mono(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl,
di(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl or .dbd.O.
[0067] The phrase "regulating mucus clearance" encompasses
controlling, promoting and/or influencing mucus clearance.
[0068] As used herein, the terms "treatment" and "treating"
encompass prophylactic administration of the compound or a
pharmaceutical composition comprising the compound ("prophylaxis")
as well as remedial therapy to reduce or eliminate a disease or
disorder mentioned herein. Prophylactic administration is intended
for preventing disorders or preventing recurrence of disorders and
may be used to treat a subject that is at risk of having or
suffering from one or more disorders mentioned herein. Thus, as
used herein, the term "treatment", or a derivative thereof,
contemplates partial or complete inhibition of the stated disease
state, when an active ingredient of the invention is administered
prophylactically or following the onset of the disease state for
which such active ingredient of the is administered. "Prophylaxis"
refers to administration of the active ingredient(s) to a mammal to
protect the mammal from any of the disorders set forth herein, as
well as others.
[0069] As used herein, the term "subject" encompasses animals,
including mammals and fish. Preferably the term refers to mammals
such as a humans, cattle and horses, more preferably to humans and
domestic animals such as cats, dogs, and horses, and most
preferably to humans.
[0070] Preferred compounds of the invention include those where
R.sub.1 is hydrogen, A is
##STR00015##
OR.sub.2 and OR.sub.3 represent a ring of formula (Ia)
##STR00016##
[0071] (Ia) where X is C.dbd.O;
Y is CH.dbd.CH, and R is substituted benzoyl or substituted
naphthoyl. These compounds are hereinafter referred to as compounds
of Formula A-1.
[0072] In compounds of Formula A-1, each benzoyl and naphthoyl is
substituted with from 1-5 independently selected R.sub.b groups.
Preferred R.sub.b groups on benzoyl and naphthoyl include
C.sub.1-C.sub.10 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8
cycloalkyl(C.sub.1-C.sub.6)alkyl, C.sub.3-C.sub.8
cycloalkyl(C.sub.1-C.sub.6)alkoxy, C.sub.1-C.sub.10 alkoxy,
halogen, hydroxy, cyano, nitro, amino,
mono(C.sub.1-C.sub.6)alkylamino, di(C.sub.1-C.sub.6)alkylamino,
C.sub.2-C.sub.10alkenyl, C.sub.2-C.sub.10alkynyl, C.sub.1-C.sub.6
haloalkyl, C.sub.1-C.sub.6 haloalkoxy, amino(C.sub.1-C.sub.6)alkyl,
mono(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl or
di(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl.
[0073] Or, any two adjacent R.sub.b groups together with the atoms
to which they are attached form a partially saturated 5-8 membered
ring, where the 5-8 membered ring is optionally substituted with
from 1 to 3 of R.sub.e, [0074] where each R.sub.e is
C.sub.1-C.sub.10 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8
cycloalkyl(C.sub.1-C.sub.6)alkyl, C.sub.3-C.sub.8
cycloalkyl(C.sub.1-C.sub.6)alkoxy, C.sub.1-C.sub.10 alkoxy,
halogen, hydroxy, cyano, nitro, amino,
mono(C.sub.1-C.sub.6)alkylamino, di(C.sub.1-C.sub.6)alkylamino,
C.sub.2-C.sub.10alkenyl, C.sub.2-C.sub.10alkynyl, C.sub.1-C.sub.6
haloalkyl, C.sub.1-C.sub.6 haloalkoxy, amino(C.sub.1-C.sub.6)alkyl,
mono(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl or
di(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl.
[0075] Preferably, R is benzoyl or naphthoyl, each of which is
substituted with 1-3 of R.sub.b, where each R.sub.b is
independently C.sub.1-C.sub.10 alkyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.3-C.sub.8 cycloalkyl(C.sub.1-C.sub.6)alkyl, C.sub.3-C.sub.8
cycloalkyl(C.sub.1-C.sub.6) alkoxy, C.sub.1-C.sub.10 alkoxy,
halogen, mono(C.sub.1-C.sub.6)alkylamino,
di(C.sub.1-C.sub.6)alkylamino, C.sub.2-C.sub.10alkenyl,
C.sub.2-C.sub.10alkynyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
haloalkoxy, mono(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl
or di(C.sub.1-C.sub.6)alkylamino(C.sub.1-C.sub.6)alkyl.
[0076] More preferably, R is benzoyl or naphthoyl, each of which is
substituted with 1-2 of R.sub.b, where each R.sub.b is
independently C.sub.1-C.sub.10 alkyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.3-C.sub.8 cycloalkyl(C.sub.1-C.sub.6)alkyl, C.sub.3-C.sub.8
cycloalkyl(C.sub.1-C.sub.6) alkoxy, C.sub.2-C.sub.10alkenyl, or
C.sub.2-C.sub.10alkynyl.
[0077] In one aspect, the invention relates to compounds of Formula
(I):
##STR00017##
wherein [0078] A is
[0078] ##STR00018## [0079] R is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.1-C.sub.6 alkyl esters,
C.sub.2-C.sub.6 alkenyl esters, amines, amides, aryl esters,
cycloalkyl esters, cycloalkenyl esters, purines, pyrimidines,
heterocycle, or heteroaryl; [0080] R.sub.1 is H or --(CO)CH.sub.3;
and [0081] R.sub.2 and R.sub.3 at each occurrence are independently
--CH.sub.2(CO)CH.sub.3, --CH.sub.2 (CO)CH.sub.2CH.sub.3, [0082]
--CH.sub.2 (CO)CH(CH.sub.3).sub.2, --CH.sub.2
(CO)CH.sub.2CH.sub.2CH.sub.3, --CH.sub.2 (CO)CH(CH.sub.3)
CH.sub.2CH.sub.3, or --CH.sub.2 (CO)CH.sub.2CH(CH.sub.3).sub.2, or
OR.sub.2 and OR.sub.3 can be taken together to form a six membered
ring of the formula (Ia)
[0082] ##STR00019## [0083] wherein X is C.dbd.O or CH(CH.sub.3);
[0084] wherein the bracketed-dashed bonds indicate attachment to
backbone; and [0085] Y is CH.dbd.CH, C.dbd.O, CHCH.sub.3, or
CH.sub.2; [0086] n is 1 or 0; and [0087] with the proviso that when
OR.sub.2 and OR.sub.3 are taken together to form a ring of the
formula (Ia), wherein X is C.dbd.O and the double bond is present;
when A is
##STR00020##
[0087] and when n is 1, R is not:
##STR00021## [0088] or a pharmaceutically acceptable salt, solvate,
hydrate, complex, or combination thereof.
[0089] In a broad aspect, R is alkyl, alkyl ester, halogen,
alkenyl, alkenyl ester, cycloalkyl, cycloalkyl ester, aryl, aryl
ester, heteroaryl, heterocycle, heterocycloalkyl or
heterocyclyl.
[0090] In another embodiment of this aspect, the compound is of
Formula (I), wherein R is
##STR00022## ##STR00023## ##STR00024## ##STR00025##
[0091] In a further embodiment of this aspect, the compound is of
Formula (I), wherein R is benzoyl, .alpha.-naphthoyl,
.beta.-naphthoyl, .alpha.-anthracoyl, .beta.-anthracoyl, or
.gamma.-anthracoyl.
[0092] In another embodiment of this aspect, OR.sub.2 and OR.sub.3
are taken together to form a ring of formula (Ia), wherein the ring
is
##STR00026##
[0093] wherein the bracketed-dashed bonds indicate the point of
attachment to the backbone.
[0094] In even another embodiment of this aspect, R.sub.2 and
R.sub.3 are each independently
##STR00027##
[0095] In a preferred embodiment, the invention provides compounds
of Formula (II):
##STR00028##
wherein [0096] A is
[0096] ##STR00029## [0097] R is C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkenyl, C.sub.1-C.sub.6 alkyl esters,
C.sub.2-C.sub.6 alkenyl esters, amines, amides, aryl esters,
cycloalkyl esters, cycloalkenyl esters, purines, pyrimidines,
heterocycle, or heteroaryl; [0098] R.sub.1 is H or --COCH.sub.3;
and [0099] R.sub.2 and R.sub.3 at each occurrence are independently
--CH.sub.2COCH.sub.3, --CH.sub.2COCH.sub.2CH.sub.3, [0100]
--CH.sub.2COCH(CH.sub.3).sub.2,
--CH.sub.2COCH.sub.2CH.sub.2CH.sub.3,
--CH.sub.2COCH(CH.sub.3)CH.sub.2CH.sub.3, or
--CH.sub.2COCH.sub.2CH(CH.sub.3).sub.2, or OR.sub.2 and OR.sub.3
can be taken together to form a six membered ring of the formula
(Ia)
[0100] ##STR00030## [0101] wherein X is C.dbd.O or CH(CH.sub.3);
[0102] wherein the bracketed-dashed bonds indicate attachment to
backbone; [0103] Y is CH.dbd.CH, C.dbd.O, or CH.sub.2; and [0104] n
is 1 or 0; and [0105] with the proviso that when OR.sub.2 and
OR.sub.3 are taken together to form a ring of the formula (Ia),
wherein X is C.dbd.O and the double bond is present; when A is
##STR00031##
[0105] and when n is 1, R is not:
##STR00032##
or a pharmaceutically acceptable salt, solvate, hydrate, complex,
or combination thereof.
[0106] In a further preferred embodiment, the compound of Formula
(I) is:
##STR00033##
[0107] In one aspect of this preferred embodiment, the compound of
Formula (I) is:
##STR00034##
[0108] In a further preferred embodiment, the compound of Formula
(I) is:
##STR00035##
[0109] In one aspect of this preferred embodiment, the compound of
Formula (I) is:
##STR00036##
[0110] In a further preferred embodiment, the compound of Formula
(I) is:
##STR00037##
[0111] In one aspect of this preferred embodiment, the compound of
Formula (I) is:
##STR00038##
[0112] The invention also relates to compounds, or pharmaceutically
acceptable salts, solvates, hydrates, complexes, or combination
thereof, of Formula (III):
##STR00039##
wherein [0113] R is H, OH, halogen, C.sub.1-C.sub.6 lower alkyl,
C.sub.1-C.sub.6 alkyl esters, C.sub.2-C.sub.6 alkenyl esters,
amino, amido, aldehydro, aryl ester, cycloalkyl ester, cycloalkenyl
ester, purinyl, pyrimidinyl, heterocyclyl, aryl, or heteroaryl;
[0114] Y is C.dbd.O, CH.dbd.CH, CHCH.sub.3 or CH.sub.2; and [0115]
n is 1 or 0.
[0116] In one embodiment of this aspect, the compound of formula
(III) is of formula (IV):
##STR00040##
wherein [0117] R is H, OH, halogen, C.sub.1-C.sub.6 lower alkyl,
C.sub.1-C.sub.6 alkyl esters, C.sub.2-C.sub.6 alkenyl ester, amino,
amido, aldehydro, aryl ester, cycloalkyl ester, cycloalkenyl ester,
purinyl, pyrimidinyl, heterocyclyl, aryl, or heteroaryl; [0118] Y
is C.dbd.O, CH.dbd.CH, CHCH.sub.3 or CH.sub.2; and [0119] n is 1 or
0.
[0120] In a preferred embodiment the compound of formula (III)
is:
##STR00041##
[0121] In one aspect of this preferred embodiment, the compound
is:
##STR00042##
[0122] In another preferred embodiment the compound of formula
(III) is:
##STR00043##
[0123] In one aspect of this preferred embodiment, the compound
is:
##STR00044##
[0124] In another preferred embodiment the compound of formula
(III) is:
##STR00045##
[0125] In one aspect of this preferred embodiment, the compound
is:
##STR00046##
[0126] In another preferred embodiment the compound of formula
(III) is:
##STR00047##
[0127] In one aspect of this preferred embodiment, the compound
is:
##STR00048##
In another preferred embodiment the compound of formula (III)
is:
##STR00049##
[0128] In one aspect of this preferred embodiment, the compound
is:
##STR00050##
[0129] In another preferred embodiment the compound of formula
(III) is:
##STR00051##
[0130] In one aspect of this preferred embodiment, the compound
is:
##STR00052##
[0131] The compounds of Formulas (I)-(IV) may have asymmetric
centers and occur as racemates, racemic mixtures and as individual
diastereomers, or enantiomers. All isomeric forms are included
within the scope of the invention. The invention also encompasses
radiolabelled forms of the compounds as well as all physical
states, i.e., liquids (oils) and solids (including amorphous forms
and crystalline forms), in which the compounds may exist.
[0132] In another aspect, the invention relates to pharmaceutical
formulations comprising a compound, or pharmaceutically acceptable
salt, solvate, hydrate, complex, or combination thereof, of any of
Formulas (I), (II), (III), or (IV) and at least one
pharmaceutically acceptable carrier, excipient, solvent, adjuvant
or diluent.
[0133] In a preferred embodiment of this aspect, the pharmaceutical
formulation comprises a compound of Formula (II) or (IV).
[0134] In another aspect, the invention provides methods for
regulating mucus transport comprising administering to a subject,
or contacting a cell with, a compound of the invention, or a
pharmaceutically acceptable salt, solvate, hydrate, complex, or
combination thereof, in an amount effective to regulate mucus
transport in the subject or cell.
[0135] In another aspect, the invention provides methods for
treating conditions or diseases related to, or associated with,
decreased mucus transport comprising administering to a subject a
compound of the invention or a pharmaceutically acceptable salt,
solvate, hydrate, complex, or combination thereof, in an amount
effective to treat the condition or disease. This method of
treating conditions or diseases associated with decreased mucus
transport can help prevent, treat, reduce the severity of, or delay
the onset or progression of symptoms and disease states associated
with decreased mucus transport. Such conditions or diseases include
the non-limiting examples of chronic airway obstruction, asthma,
pulmonary disease, pulmonary infection, and cystic fibrosis.
[0136] In one embodiment, the method of treatment can be used to
treat chronic airway obstruction.
[0137] In one embodiment, the method of treatment can be used to
treat asthma.
[0138] In one embodiment, the method of treatment can be used to
treat pulmonary diseases such as emphysema, pulmonary fibrosis,
and/or smokers cough.
[0139] In one embodiment, the method of treatment can be used to
treat pulmonary infection, including, but not limited to,
pneumonia, or Pseudomonas.
[0140] In a preferred embodiment, the method of treatment can be
used to where the disease is cystic fibrosis.
[0141] In another aspect, the invention provides methods for
treating the symptoms related to conditions or diseases associated
with decrease mucus transport, comprising administering to a
subject in need of such treatment, a therapeutically effective
amount of a compound of the invention, or a pharmaceutically
acceptable salt, solvate, hydrate, complex, or combination
thereof.
[0142] In another aspect, the invention provides methods for
treating conditions or diseases associated with mucociliary
dysfunction, comprising administering to a subject in need of such
treatment, a therapeutically effective amount of a compound of the
invention, or a pharmaceutically acceptable salt, solvate, hydrate,
complex, or combination thereof.
[0143] In another aspect, the invention provides methods for
treating the symptoms related to conditions or diseases associated
with mucociliary dysfunction, comprising administering to a subject
in need of such treatment, a therapeutically effective amount of a
compound of the invention, or a pharmaceutically acceptable salt,
solvate, hydrate, complex, or combination thereof.
[0144] In another embodiment of this aspect, the method can
optionally comprise in combination with the compound of Formula
(I)-(IV) or a pharmaceutically acceptable salt, solvate, hydrate,
complex, or combination thereof, an effective amount of a compound
known to be useful for the treatment of conditions or diseases
associated with decreased mucus transport. The methods of the
invention can optionally comprise additional therapeutic regimen
such as supportive or adjuvant therapy.
[0145] In one embodiment of the methods of the invention, the
subject is a mammal. In a more preferred embodiment, the mammal is
a human.
[0146] The methods of the invention employ therapeutically
effective amounts: for inhalation, oral, parenteral, sublingual,
intranasal, intrathecal, depo, implants, topical, and rectal
administration from about 0.1 pg/day to about 100 mg/day. The
therapeutically effective amounts will vary according to various
parameters including, for example, the route of administration, the
distribution of the compound, the metabolism of the compound, the
excretion of the compound, the particular therapeutic use, and the
physical characteristics of the subject/patient, and are well
within the knowledge of those skilled in the art.
[0147] In a preferred aspect, the therapeutically effective amounts
for oral non-inhalation administration is about 1 mg/day to about
100 mg/day
[0148] In a preferred aspect, the therapeutically effective amounts
for parenteral, and depo administration is from about 1 .mu.g/day
to about 100 mg/day.
[0149] In a preferred aspect, the therapeutically effective amounts
for inhalation administration is about 0.1 pg/day to about 1
.mu.g/day.
[0150] The invention also includes the use of a compound of Formula
(I)-(IV), or a pharmaceutically acceptable salt, solvate, hydrate,
complex, or combination thereof for the manufacture of a medicament
for use in treating a subject who has, or in preventing a subject
from developing, disorders or diseases associated with decreased
mucus transport or mucociliary dysfunction, and symptoms associated
with those disorders or diseases, and who is in need of such
treatment.
[0151] In one aspect, this use of a compound of formula (I)-(IV)
can be employed where the disease or condition is chronic airway
obstruction.
[0152] In another aspect, this use of a compound of formula
(I)-(IV) can be employed where the disease or condition is
asthma.
[0153] In another aspect, this use of a compound of formula
(I)-(IV) can be employed where the disease or condition is
pulmonary disease.
[0154] In another aspect, this use of a compound of formula
(I)-(IV) can be employed where the disease or condition is
pulmonary infection.
[0155] In another aspect, this use of a compound of formula
(I)-(IV) can be employed where the disease or condition is cystic
fibrosis.
[0156] In another aspect, compounds of the invention can be
employed for treating a industrial related disease or condition
caused or exacerbated by inhaling gases, particles of textiles,
grit, or other industrial particles or fumes. Specific examples of
particles and grit include, for example, iron oxides, silica, talc,
carbon, graphite, fibers, wood dust, grain dust, organic solvents
and pollutant gases.
[0157] In still another aspect, the compounds of the invention can
be employed for treating a disease or condition resulting from
inhalation of bacterial or other pathogenic particles, e.g., fungal
particles. Thus, the invention also encompasses methods of clearing
pathogenic particles, such as particles that comprise bacteria,
e.g., anthrax or fungus particles.
[0158] The invention also includes a container kit including a
plurality of containers, each container including one or more unit
dose of a compound of formula (I)-(IV), or a pharmaceutically
acceptable salt, solvate, hydrate, complex, or combination
thereof.
[0159] In an embodiment, this container kit includes each container
adapted for oral delivery and includes an inhaler, tablet, gel, or
capsule.
[0160] In an embodiment, this container kit includes each container
adapted for parenteral delivery and includes a depot product,
syringe, ampoule, or vial.
[0161] In an embodiment, this container kit includes each container
adapted for topical delivery and includes a patch, medipad,
ointment, or cream.
[0162] The compounds of formula (I) can form salts when reacted
with appropriate acids or bases. Pharmaceutically acceptable salts
are generally preferred over the corresponding compounds of formula
(I) since they frequently produce compounds that are usually more
water soluble, stable and/or more crystalline. Pharmaceutically
acceptable salts are any salt which retains the activity of the
parent compound and does not impart any deleterious or undesirable
effect on the subject to whom it is administered and in the context
in which it is administered. Pharmaceutically acceptable salts
include acid addition salts of both inorganic and organic acids.
Preferred pharmaceutically acceptable salts include salts such as
those described by Berge, Bighley, and Monkhouse, J. Pharm. Sci.,
1977, 66, 1-19. Such salts may be formed from inorganic and organic
acids. Representative examples thereof include maleic, fumaric,
benzoic, ascorbic, pamoic, succinic, bismethylenesalicylic,
methanesulfonic, ethanedisulfonic, acetic, propionic, tartaric,
salicylic, citric, gluconic, aspartic, stearic, palmitic, itaconic,
glycolic, p-aminobenzoic, glutamic, benzenesulfonic, hydrochloric,
hydrobromic, sulfuric, cyclohexylsulfamic, phosphoric and nitric
acids. For other acceptable salts, see Int. J. Pharm., 33, 201-217
(1986). The compounds of formula (I) can also form solvates,
hydrates, complexes, or combination thereof.
METHODS OF THE INVENTION
[0163] The compounds of the invention, pharmaceutical formulations
comprising said compounds, and pharmaceutically acceptable salts
thereof, are useful for treating a subject, preferably a mammal,
more preferably a human, suffering from a disease or condition
associated with decreased mucus transport, and are useful for
helping to prevent or delay the onset of such a disease or
condition. The compounds and formulations of the invention are
particularly useful for treating, preventing, or slowing the
progression of chronic airway obstruction, asthma, pulmonary
disease, pulmonary infection, and cystic fibrosis. When treating or
preventing a disease and condition associated with decreased mucus
transport, and the associated symptoms, the compounds of the
invention can either be used individually or in combination, as is
best for the subject.
[0164] With regard to these diseases and conditions, the term
"treating" means that compounds of the invention can be used in
subjects, preferably human subjects/patients, with existing
condition or disease. The compounds of the invention will not
necessarily cure the subject who has the disease but will delay or
slow the progression or prevent further progression of the disease
thereby giving the individual a more useful life span.
[0165] The term "preventing" means that that if the compounds of
the invention are administered to those who do not now have the
disease, or symptom(s) of the condition, but who would normally
develop the disease or be at increased risk for the disease, they
will not develop the disease. In addition, "preventing" also
includes delaying the development of the disease in an individual
who will ultimately develop the disease or would be at risk for the
disease due to age, familial history, genetic or chromosomal
abnormalities, and/or due to the presence of one or more biological
markers for the disease. By delaying the onset of the disease,
compounds of the invention can prevent the individual from getting
the disease during the period in which the individual would
normally have gotten the disease or reduce the rate of development
of the disease or some of its effects but for the administration of
compounds of the invention up to the time the individual ultimately
gets the disease. Preventing also includes administration of the
compounds of the invention to those individuals thought to have
predisposition for the disease.
[0166] In a preferred aspect, the compounds of the invention are
useful for slowing the progression of disease symptoms.
[0167] In another preferred aspect, the compounds of the invention
are useful for preventing the further progression of disease
symptoms.
[0168] In treating or preventing the above diseases, the compounds
of the invention are administered in a therapeutically effective
amount. The therapeutically effective amount will vary depending on
the particular compound used, the physical characteristics of the
subject to be treated, and the route of administration, as is known
to those skilled in the art.
[0169] In treating a subject displaying any of the diagnosed above
conditions a physician may administer a compound of the invention
immediately and continue administration indefinitely, as
needed.
Dosage Forms and Amounts
[0170] The compounds of the invention can be administered orally,
parenterally, (IV, IM, depo-IM, SQ, and depo SQ), sublingually,
intranasally, by inhalation, intrathecally, topically, vaginally,
or rectally. Dosage forms known to those of skill in the art are
suitable for delivery of the compounds of the invention.
[0171] Compositions are provided that contain therapeutically
effective amounts of the compounds of the invention. The compounds
are preferably formulated into suitable pharmaceutical preparations
such as aerosols, inhalants, tablets, capsules, or elixirs for oral
administration or in sterile solutions or suspensions for
parenteral administration. Typically the compounds described above
are formulated into pharmaceutical compositions using techniques
and procedures well known in the art.
[0172] About 0.1 pg to about 100 mg of a compound or mixture of
compounds of the invention or a physiologically acceptable salt,
solvate, hydrate, complex, ester, or combination thereof, is
compounded with a physiologically acceptable vehicle, carrier,
excipient, binder, preservative, stabilizer, flavor, etc., in a
unit dosage form as called for by accepted pharmaceutical practice.
The amount of active substance in those compositions or
preparations is such that a suitable dosage in the range indicated
is obtained. The compositions are preferably formulated in a unit
dosage form, each dosage containing from about 0.1 pg to about 100
mg, preferably about 1 pg to about 1 .mu.g for inhalation
administration, preferably about 100 ng to about 1 mg for
injection/intravenous administration, or about 1 mg to about 100 mg
for oral administration (e.g., tablets, elixirs, capsules, etc.),
of the active ingredient. The term "unit dosage from" refers to
physically discrete units suitable as unitary dosages for human
subjects and other mammals, each unit containing a predetermined
quantity of active material calculated to produce the desired
therapeutic effect, in association with a suitable pharmaceutical
excipient.
[0173] To prepare pharmaceutical compositions, one or more
compounds of the invention are mixed with a suitable
pharmaceutically acceptable carrier. Upon mixing or addition of the
compound(s), the resulting mixture may be a solution, suspension,
emulsion, or the like. Liposomal suspensions may also be suitable
as pharmaceutically acceptable carriers. These may be prepared
according to methods known to those skilled in the art. The form of
the resulting mixture depends upon a number of factors, including
the intended mode of administration and the solubility of the
compound in the selected carrier or vehicle. The effective
concentration is sufficient for lessening or ameliorating at least
one symptom of the disease, disorder, or condition treated and may
be empirically determined.
[0174] Pharmaceutical carriers or vehicles suitable for
administration of the compounds provided herein include any such
carriers known to those skilled in the art to be suitable for the
particular mode of administration. In addition, the active
materials can also be mixed or blended with other active materials
that do not impair the desired action, or with materials that
supplement the desired action, or have another action. The
compounds may be formulated as the sole pharmaceutically active
ingredient in the composition or may be combined with other active
ingredients.
[0175] Where the compounds exhibit insufficient solubility, methods
for solubilizing may be used. Such methods are known and include,
but are not limited to, using cosolvents such as dimethylsulfoxide
(DMSO), using surfactants such as Tween.RTM., and dissolution in
aqueous sodium bicarbonate. Derivatives of the compounds, such as
salts, solvates, hydrates, complexes, or prodrugs may also be used
in formulating effective pharmaceutical compositions.
[0176] The concentration of the compound is effective for delivery
of an amount upon administration that lessens or ameliorates at
least one symptom of the poisoning or disorder for which the
compound is administered. Typically, the compositions are
formulated for single dosage administration.
[0177] The compounds of the invention may be prepared with carriers
that protect them against rapid elimination from the body, such as
time-release formulations or coatings. Such carriers include
controlled release formulations, such as, but not limited to,
microencapsulated delivery systems. The active compound is included
in the pharmaceutically acceptable carrier in an amount sufficient
to exert a therapeutically useful effect in the absence of
undesirable side effects on the subject treated. The
therapeutically effective concentration may be determined
empirically by testing the compounds in known in vitro and in vivo
model systems for the treated disorder.
[0178] The compounds and compositions of the invention can be
enclosed in multiple or single dose containers. The enclosed
compounds and compositions can be provided in kits, for example,
including component parts that can be assembled for use. For
example, a compound inhibitor in lyophilized form and a suitable
diluent may be provided as separated components for combination
prior to use. A kit may include a compound inhibitor and a second
therapeutic agent for co-administration. The inhibitor and second
therapeutic agent may be provided as separate component parts. A
kit may include a plurality of containers, each container holding
one or more unit dose of the compound of the invention. The
containers are preferably adapted for the desired mode of
administration, including, but not limited to tablets, gel
capsules, sustained-release capsules, and the like for oral
administration; depot products, pre-filled syringes, ampoules,
vials, and the like for parenteral administration; and patches,
medipads, creams, and the like for topical administration.
[0179] The concentration of active compound in the drug composition
will depend on absorption, route of administration, metabolism,
inactivation, and excretion rates of the active compound, the
dosage schedule, and amount administered as well as other factors
known to those of skill in the art.
[0180] The active ingredient may be administered at once, or may be
divided into a number of smaller doses to be administered at
intervals of time. It is understood that the precise dosage and
duration of treatment is a function of the disease being treated
and may be determined empirically using known testing protocols or
by extrapolation from in vivo or in vitro test data. It is to be
noted that concentrations and dosage values may also vary with the
severity of the condition to be alleviated. It is to be further
understood that for any particular subject, specific dosage
regimens should be adjusted over time according to the individual
need and the professional judgment of the person administering or
supervising the administration of the compositions, and that the
concentration ranges set forth herein are exemplary only and are
not intended to limit the scope or practice of the claimed
compositions.
[0181] If oral, non-inhalation administration is desired, the
compound should be provided in a composition that protects it from
the acidic environment of the stomach. For example, the composition
can be formulated in an enteric coating that maintains its
integrity in the stomach and releases the active compound in the
intestine. The composition may also be formulated in combination
with an antacid or other such ingredient.
[0182] Oral compositions will generally include an inert diluent or
an edible carrier and may be compressed into tablets or enclosed in
gelatin capsules. For the purpose of oral therapeutic
administration, the active compound or compounds can be
incorporated with excipients and used in the form of tablets,
capsules, or troches. Pharmaceutically compatible binding agents
and adjuvant materials can be included as part of the
composition.
[0183] The tablets, pills, capsules, troches, and the like can
contain any of the following ingredients or compounds of a similar
nature: a binder such as, but not limited to, gum tragacanth,
acacia, corn starch, or gelatin; an excipient such as
microcrystalline cellulose, starch, or lactose; a disintegrating
agent such as, but not limited to, alginic acid and corn starch; a
lubricant such as, but not limited to, magnesium stearate; a
gildant, such as, but not limited to, colloidal silicon dioxide; a
sweetening agent such as sucrose or saccharin; and a flavoring
agent such as peppermint, methyl salicylate, or fruit
flavoring.
[0184] When the dosage unit form is a capsule, it can contain, in
addition to material of the above type, a liquid carrier such as a
fatty oil. In addition, dosage unit forms can contain various other
materials, which modify the physical form of the dosage unit, for
example, coatings of sugar and other enteric agents. The compounds
can also be administered as a component of an elixir, suspension,
syrup, wafer, chewing gum or the like. A syrup may contain, in
addition to the active compounds, sucrose as a sweetening agent and
certain preservatives, dyes and colorings, and flavors.
[0185] The active materials can also be mixed or blended with other
active materials that do not impair the desired action, or with
materials that supplement the desired action.
[0186] Solutions or suspensions used for parenteral, intradermal,
subcutaneous, or topical application can include any of the
following components: a sterile diluent such as water for
injection, saline solution, fixed oil, a naturally occurring
vegetable oil such as sesame oil, coconut oil, peanut oil,
cottonseed oil, and the like, or a synthetic fatty vehicle such as
ethyl oleate, and the like, polyethylene glycol, glycerine,
propylene glycol, or other synthetic solvent; antimicrobial agents
such as benzyl alcohol and methyl parabens; antioxidants such as
ascorbic acid and sodium bisulfite; chelating agents such as
ethylenediaminetetraacetic acid (EDTA); buffers such as acetates,
citrates, and phosphates; and agents for the adjustment of tonicity
such as sodium chloride and dextrose. Parenteral preparations can
be enclosed in ampoules, disposable syringes, or multiple dose
vials made of glass, plastic, or other suitable material. Buffers,
preservatives, antioxidants, and the like can be incorporated as
required.
[0187] Where administered intravenously, suitable carriers include
physiological saline, phosphate buffered saline (PBS), and
solutions containing thickening and solubilizing agents such as
glucose, polyethylene glycol, polypropyleneglycol, and mixtures
thereof. Liposomal suspensions including tissue-targeted liposomes
may also be suitable as pharmaceutically acceptable carriers. These
may be prepared according to methods known for example, as
described in U.S. Pat. No. 4,522,811.
[0188] The active compounds may be prepared with carriers that
protect the compound against rapid elimination from the body, such
as time-release formulations or coatings. Such carriers include
controlled release formulations, such as, but not limited to,
implants and microencapsulated delivery systems, and biodegradable,
biocompatible polymers such as collagen, ethylene vinyl acetate,
polyanhydrides, polyglycolic acid, polyorthoesters, polylactic
acid, and the like. Methods for preparation of such formulations
are known to those skilled in the art.
[0189] The compounds of the invention can be administered by
inhalation, orally or intranasally, parenterally (IV, IM, depo-IM,
SQ, and depo-SQ), sublingually, intrathecally, topically, or
rectally. Dosage forms known to those skilled in the art are
suitable for delivery of the compounds of the invention.
[0190] Compounds of the invention may be administered enterally or
parenterally. When administered orally, compounds of the invention
can be administered in usual dosage forms for oral administration
as is well known to those skilled in the art. These dosage forms
include the usual solid unit dosage forms of tablets and capsules
as well as liquid dosage forms such as solutions, suspensions, and
elixirs. When the solid dosage forms are used, it is preferred that
they be of the sustained release type so that the compounds of the
invention need to be administered only once or twice daily.
[0191] The oral dosage forms are administered to the subject 1, 2,
3, or 4, or as needed, times daily. It is preferred that the
compounds of the invention be administered either three or fewer
times, more preferably once or twice daily. Hence, it is preferred
that the compounds of the invention be administered in oral dosage
form. It is preferred that whatever oral dosage form is used, that
it be designed so as to protect the compounds of the invention from
the acidic environment of the stomach. Enteric coated tablets are
well known to those skilled in the art. In addition, capsules
filled with small spheres each coated to protect from the acidic
stomach, are also well known to those skilled in the art.
[0192] In a preferred embodiment, the compounds of the invention
are administered in an inhalant form.
[0193] As noted above, depending on whether asymmetric carbon atoms
are present, the compounds of the invention can be present as
mixtures of isomers, as racemates, or in the form of pure
isomers.
[0194] Salts of compounds are preferably the pharmaceutically
acceptable or non-toxic salts of compounds of formula I. For
isolation and purification purposes it is also possible to use
pharmaceutically unacceptable salts. [0195] Synthesis of
Compounds
[0196] Various synthetic methodologies can be used to make
compounds of the invention; certain of the brevetoxins are suitable
starting materials. Suitable methodologies are known in the art.
Representative synthetic procedures for preparing compounds of the
invention from such starting materials are disclosed in, e.g.,
Mende, T. J., et al., Tetr. Lett., 1990; 31(37):5307-5310; Trainer,
V. L., et al., Molec. Pharm., 1991; 40(6):988-994; Keck, G. E., et
al., Tetrahedron Lett., 1987, 28:139-142; Alvarez, E., et al.,
Chem. Rev., 1995, 95:1953-1980; Rein, et al., 1994: (a) J. Org
Chem., 59:2107-2113; (b) J. Org. Chem. 59:2101-2106. Each of these
references is incorporated herein by reference in its entirety.
Those skilled in the art will appreciate that minor modifications
can be made to the particular procedures to arrive at compounds of
the invention.
[0197] The following examples serve merely to illustrate the
invention and should not be viewed to limit the invention in scope
or spirit.
EXAMPLES
[0198] General. All solvents used were HPLC grade. Brevetoxins were
purified from laboratory cultures of the algae Karenia brevis (also
called Ptychodiscus brevis and Gymnodinium breve) by a combination
of chloroform/methanol extraction and TLC. Brevetoxin can be
isolated and purified from native sources, such as K. brevis, or
other red tide organisms. Suitable purification methodologies are
well known in the art. Preferably, brevetoxins are extracted from
K. brevis cultures. This algae is available from the Provasoli
Guillard National Center for Culture of Marine Phytoplankton, West
Boothbay Harbor, Me., as strain number CCMP718. In addition, the
synthesis of Brevetoxin B has been reported: J. Am. Chem. Soc.,
117, 1171 (K. C. Nicolaou et al., 1995).
[0199] Starting materials (PbTx-2, -3, and -9) and products were
routinely purified by reversed phase HPLC (85% isocratic methanol)
using a Microsorb-MV, C-18 column (5 um, 25-cm bed) and monitored
by UV at 215 or 195 nM and/or refractive index. Proton NMR spectra
were recorded in CDCl3 (CHC13 internal standard) at 400 MHz. Mass
spectra were run in either DCI or FAB mode. High-resolution mass
spectra were obtained from the mass spectrometry facility at the
University of California, Riverside.
Synthesis of Brevetoxin Derivatives
[0200] A tenfold excess (relative to PbTx-3) of carbonyl
diimidazole and the corresponding acid (benzoic, (.alpha.-naphthoic
or .beta.-naphthoic) were combined under nitrogen, at room
temperature, in dry benzene. The solution was stirred for 30 min
and then added to PbTx-3 in a 5 ml reaction vial. The reaction vial
was sealed and the mixture was stirred overnight at 80.degree. C.
The reaction mixture was washed with an equal volume (3.times.) of
saturated sodium bicarbonate, an equal volume (3.times.) of 10% HCl
and evaporated under vacuum. The residue was purified using
HPLC.
[0201] Benzoyl-PbTx-3 (1). Diagnostic peaks in the 1H NMR include
87.44 (2H, t, J=7.2 Hz), 7.56 (1H, t, J=7.2 Hz), 8.061 (2H, d,
J=8.4 Hz), 4.81 (2H, dd, J=5.2 Hz) (C42). The C42 methylene is
typically shifted downfield from its position in PbTx-3, and these
diastereotopic protons are split into a doublet of doublets in the
esters, whereas they appear as a singlet in PbTx-3. DCI MS (NH3):
1002 (M+1). HRMS (FAB): calc'd for CS7H, O, S (MH+), calc'd
1001.5262, found 1001.5287.
[0202] .alpha.-Naphthoyl-PbTx-3 (2). Diagnostic peaks in the 'H NMR
include 88.93 (1H, d, J=8.8 Hz), 8.23 (1H, d, J=8.8 Hz), 8.03 (1H,
d, J=8.8 Hz), 7.83 (1H, d, J=8.8 Hz), 7.64 (1H, t, J=8.8 Hz), 7.44
(2H, m), 4.92 (2H, dd, J=5.2 Hz) DCI MS (NH3): 1052 (M+1). HRMS
(FAB): calc'd for C6iH, 90, 5 (MH+), calc'd 1051.5419, found
1051.5367.
[0203] .beta.-Naphthoyl-Pb7x-3 (3). Diagnostic peaks in the 1H NMR
include 88.64 (1H, s), 7.95 (3H, m), 7.58 (3H, m) 4.89 (1H, s).
Example 1
[0204] Oxidation of C-42 of PbTx-2 to Ester (1). PbTx-2 (4.8 mg,
5.37 uM) was oxidized with activated MnO.sub.2, to the
corresponding methyl ester 1, via the cyanohydrin, according to the
procedure described by Corey. The reaction mixture was filtered
through Celite and concentrated in vacuo. The residue was taken up
in water (15 mL) and extracted with ether (3.times.15 mL). The
ether phase was evaporated in vacuo and the residue purified by
HPLC to yield 3.624 mg (73%) of the desired product. DCI MS
(NH.sub.3): 925 (M+1), 942 (M+NH4), 906 (M-H20).
Example 2
[0205] Hydrolysis of Methyl Ester 1 To Provide Carboxylic Acid (2).
Methyl ester 1 (3.759 mg, 4.068 uM) was dissolved in 2 mL of
THF/H.sub.20 (50:50). An aqueous solution of KOH (0.4 mL, 10 mg/mL)
was added, and the reaction mixture was stirred at ambient
temperature for 2 days. Water (1 mL) was added and the mixture
extracted with ether (3.times.2 mL). The aqueous phase was
acidified with 10% HCl and extracted with ethyl acetate (3.times.2
mL) and the organic phase evaporated in vacuo. This residue
consisted of a mixture of two products. On the basis of NMR data,
these two products appear to be the C-42 carboxylic acid with an
intact A-ring lactone and the C-42 carboxylic acid with a
hydrolyzed A-ring lactone. The residue was taken up in THF (1 mL),
and a catalytic amount of p-toluenesulfonic acid was added. The
mixture was stirred for 1 h and then evaporated in vacuo. The
residue was taken up in water (1 mL) and extracted with ethyl
acetate (3.times.2 mL). Evaporation of the solvent in vacuo yielded
3.469 mg (93%) of the crude product. Purification of the residue by
HPLC provided 1.019 mg (28%) of 2. IHNMR. DCI MS (NH3): 911 (M+1),
929 (M+NH4), 892 (M-H20).
##STR00053##
Example 3
[0206] Reduction of the C-2, C-3 Double Bond of PbTx-3 to Provide
(3). Following the procedure of Hudlicky, PbTx-3 (4.00 mg, 4.46 uM)
and Mg (approximately 200 mg, 99.98% from Timminco metals, Haley,
Ontario, Canada) were dried in vacuo over P206 for 18 h. Methanol
(3 mL freshly distilled from CaH.sub.2) was added and the mixture
stirred at room temperature (cooling in an ice bath was required)
under nitrogen for 2 h. HCl (10%, 10 mL) was added to dissolve the
magnesium methoxide and the remaining magnesium. The reaction
mixture was concentrated to approximately 5 mL and extracted with
ether (3.times.15 mL). The ether phase was evaporated in vacuo and
the residue purified by HPLC. The isolated material consisted of
the desired product with the C-2, C-3 double bond reduced and a
second product in which the double bond was reduced and the lactone
opened to the methyl ester, as evidenced by mass spectral and NMR
data. This mixture was taken up in THF, and a small amount of
p-TsOH acid was added. The mixture was stirred for 1 h and then
evaporated in vacuo. The residue was taken up in water (1 mL) and
extracted with ethyl acetate (3.times.2 mL). Evaporation in vacuo
yielded 3.256 mg (81%) of 3. DCI MS (NH3): 899 (M+
[0207] 1), 916 (M+NH), 880 (M-H20). HRMS (DCI): calcd for
Cr,oH74014 (MH+) 899.5156, found 899.5128.
Example 4
[0208] Reduction of the C-2, C-3 Double Bond of PbTx-9 To Provide
(4). PbTx-9 (7.49 mg, 8.34 AM) was reduced according to the
procedure described for the preparation of 3 to yield 1.645 mg
(22%) of 4. The 1H NMR spectrum is shown in the supplementary
material. DCI MS (NH3): 901 (M+1), 918 (M+NH4), 882 (M-H, O. HRMS
(DCI): calcd for C6pHqg014 (MH+) 901.5313, found 901.5323.
##STR00054##
Example 5
[0209] Sodium Borohydride Reduction of PbTx-3 to Form (5) and (6).
PbTx-3 (3.451 mg, 3.85 uM) was dissolved in 2.5 mL of EtOH. A large
excess of NaBH.sub.4 (5 mg) was added in one portion. The reaction
mixture was stirred at ambient temperature for 18 h. The excess
NaBH.sub.4 was decomposed by the careful addition of 10% HCl. The
reaction mixture was concentrated in vacuo to 1 mL and extracted
with CH.sub.2Cl.sub.2 (3.times.2 mL). The combined organic phases
were then evaporated to dryness, and the residue was purified by
HPLC. Two peaks were collected from the HPLC. The first peak was
the minor product 5, 0.755 mg (22%), and the second peak was the
major product 6, 1.042 mg (30%). Compound 5. DCI MS (NH3): 880,
729. FAB MS (m-nitrobenzyl alcohol matrix): 901 (M+1). HRMS (FAB):
calcd for C6pHyg014 (MH+) 901.5313, found 901.5324. Compound 6. FAB
MS (m-nitrobenzyl alcohol matrix): 903 (M+1), 766, 731. HRMS (FAB):
calcd for CrOH78014 (MH+) 903.5470, found 903.5418.
Example 6
[0210] Catalytic Reduction of PbTx-3 to Yield (7). PbTx-3 (1.8 mg,
2.00 wM) was dissolved in i-PrOH (1 mL). Acetic acid (50 AL) and a
catalytic amount of 10% Pd on activated carbon were added. The
reaction mixture was stirred at ambient temperature under an
atmosphere of H.sub.2 for 24 h. The suspension was filtered through
Celite and concentrated in vacuo to provide 0.986 mg (54%) of 7
which was not purified further. DCI MS (NH3): 903 (M+1), 920
(M+NH4), 894 (M-H20). HRMS (DCI): calcd for CrOH.8014 (MH+)
903.5470, found 903.5444.
Example 7
[0211] Epoxidation of the C-27, C-28 Double Bond of PbTx-2 To
Provide PbTx-6. Dimethyldioxirane was generated in a distillation
apparatus, connected to a dry ice condenser, according to the
procedure described by Adam17 for a small-scale preparation. The
receiving flask was charged with PbTx-2 (2.33 mg) in 5.0 mL of
acetone and was cooled in an ice/salt bath. The reaction was
monitored by HPLC. When all of the PbTx-2 was consumed, the acetone
was evaporated in vacuo, and the residue taken up in 1.0 mL of
methanol and purified by HPLC to provide 2.25 mg (95%) of PbTx-6.
DCI MS (NH.sub.3): 911 (M+1), 928 (M+NH.sub.4), 893 (M-H.sub.20).
1H and 13C NMR were identical to that reported by Shimizu.
Example 8
Airway Merchanics Experimental Protocols
[0212] Measurement of Airway Mechanics--Unsedated sheep are
restrained in a cart in the prone position with their heads
immobilized. After topical anesthesia of the nasal passages with 2%
lidocaine solution, a balloon catheter is advanced through one
nostril into the lower esophagus. The animals are intubated with a
cuffed endotracheal tube through the other nostril using a flexible
fiber optic bronchoscope. Pleural pressure is estimated with the
esophageal balloon catheter (filled with one ml of air) which is
positioned 5-10 cm from the gastroesophageal junction. In this
position the end expiratory pleural pressure ranges between -2 and
-5 cm H.sub.20. Once the balloon is placed, it is secured so that
it remains in position for the duration of the experiment. Lateral
pressure in the trachea is measured with a sidehole catheter (inner
dimension, 2.5 mm) advanced through and positioned distal to the
tip of the endotracheal tube. Transpulmonary pressure, the
difference between tracheal and pleural pressure, is measured with
a differential pressure transducer catheter system which shows no
phase shift between pressure and flow up to a frequency of 9 Hz.
For the measurement of pulmonary resistance (R.sub.L), the proximal
end of the endotracheal tube is connected to a pneumotachograph
(Fleisch, Dyna Sciences, Blue Bell, Pa.). The signals of flow and
transpulmonary pressure are recorded on an oscilloscope recorder
which is linked to a computer for on-line calculation of R.sub.L
from transpulmonary pressure, respiratory volume (obtained by
digital integration) and flow by the iso-volume technique. Analysis
of 5-10 breaths is used for the determination of R.sub.L (Abraham
et al., 1994).
[0213] Aerosol Delivery Systems--All aerosols are generated using a
disposable medical nebulizer (Raindrop.RTM., Puritan Bennett,
Lenexa, Kans.) that provide an aerosol with a mass median
aerodynamic diameter of 3.2 .mu.m (geometric SD 1.9) as determined
by an Andersen cascade impactor. The nebulizer is connected to a
dosimeter system, consisting of a solenoid valve and a source of
compressed air (20 psi). The output of the nebulizer was directed
into a plastic T-piece, one end of which is connected to the
inspiratory port of a Harvard respirator. The solenoid valve is
activated for one second at the beginning of the inspiratory cycle
of the respirator.
[0214] Aerosols were delivered at a tidal volume of 500 ml and a
rate of 20 breaths per minute (Abraham et al., 1994).
[0215] Airway Responsiveness--To assess airway responsiveness, we
perform cumulative dose response curves to carbachol by measuring
R.sub.L immediately after inhalation of buffer and after each
consecutive administration of 10 breaths of increasing
concentrations of carbachol (0.25, 0.5, 1.0, 2.0 and 4.0% w/v
buffered saline). The provocation test is discontinued when R.sub.L
increased over 400% from the post-saline value or after the highest
carbachol concentration has been administered. Airway
responsiveness is estimated by determining the cumulative carbachol
dose in breath units (BU) that increases R L by 400% (PC400) by
interpolation from the dose response curve. One breath unit (BU) is
defined as 1 breath of an aerosol solution containing 1% wt/vol
carbachol (Abraham et al., 1994).
[0216] Nasal Airway Resistance--Nasal airway resistance (NAR) is
measured with a modified mask rhinomanometry technique. The sheep's
head is placed in a plexiglass hood with attachments for a
faceplate containing a pneumotachograph to measure flow and two
catheter ports to measure the pressure differential between nose
and mouth pressure (Abraham et al., 1998).
[0217] Tracheal Mucus Velocity--Sheep are nasally intubated with an
endotracheal tube 7.5 cm in diameter shortened by 6 cm., after
topical anesthesia of the nasal passages with 2% lidocaine
solution. The cuff of the tube is placed just below the vocal cords
(verified by fluoroscopy) in order to allow for maximal exposure of
the tracheal surface area. TMV is measured in vivo by a
roentgenographic technique. Between 10 and 20 radiopaque
Teflon/bismuth trioxide disks, 1-mm diameter, 0.8-mm thick and 1.8
mg in weight, are insufflated into the trachea via the endotracheal
tube. The cephalad-axial velocities of the individual disks are
recorded on videotape from a portable image intensifier unit.
Individual disk velocities are calculated by measuring the distance
traveled by each disk during a 1-min observation period. For each
run, the mean value of all individual disk velocities is
calculated. A collar containing radiopaque reference markers of
known length are worn by the sheep, and used as a standard to
correct for magnification effects inherent in the fluoroscopy unit
(O'Riordan et al., 1997).
[0218] Statistical Analysis--If the data are normally distributed,
then parametric statistics are used; if data do not conform to a
normal distribution, non-parametric statistics are used. The basic
statistical tests include analysis of variance (ANOVA), i.e.
one-way ANOVA or two-way ANOVA with repeated measures for
multipoint analysis, and unpaired or paired t-test for the
appropriate single point analysis. The non-parametric counterparts
of these tests are: a) the Mann-Whitney test, which is the
counterpart of the unpaired t-test; b) Wilcoxon's signed ranks
test, the counterpart of the paired t-test; c) Friedman's Analysis
of Variance for related samples, i.e. randomized blocked design; d)
the Quade test, also a randomized block design test but for use
with small blocks (n.ltoreq.4); e) the Kruskal-Wallis test, ANOVA
for unrelated samples; and f) a non-parametric pairwise comparison,
analogous to the parametric Newman-Kuels pairwise test. Where
applicable, linear regression analysis is performed by method of
least squares, and correlations will be tested for with Spearman's
rho test. For all studies, significance is accepted with p<0.05
on a two tailed analysis (Conover, 1980).
[0219] The invention has been described with reference to various
specific and preferred embodiments and techniques. However, it
should be understood that many variations and modifications may be
made while remaining within the spirit and scope of the
invention.
* * * * *