U.S. patent application number 09/865160 was filed with the patent office on 2001-11-22 for methods for controlling gram negative bacteria in mammals.
This patent application is currently assigned to Millennium Pharmaceuticals, Inc.. Invention is credited to Berkowitz, Barry, Blackburn, Chris, Sachs, George.
Application Number | 20010044463 09/865160 |
Document ID | / |
Family ID | 25421781 |
Filed Date | 2001-11-22 |
United States Patent
Application |
20010044463 |
Kind Code |
A1 |
Berkowitz, Barry ; et
al. |
November 22, 2001 |
Methods for controlling gram negative bacteria in mammals
Abstract
Methods for controlling growth of gram negative bacteria, such
as Helicobacter pylori, which are agents associated with disorders
of the gastrointestinal tract of a mammal are described. They
include administering a therapeutically effective amount of a
polyether ionophore antibiotic to a mammal, such that growth of
gram negative bacteria which are agents associated with disorders
of the gastrointestinal tract of a mammal is controlled. Packaged
pharmaceutical compositions for controlling gram negative bacteria
are also described. The packaged compositions include a container
holding a therapeutically effective amount of a pharmaceutical
composition for controlling gram negative bacteria which are agents
associated with disorders of the gastrointestinal tract of a mammal
and instructions for using the pharmaceutical composition for
control of the gram negative bacteria, such that growth of gram
negative bacteria which are agents associated with disorders of the
gastrointestinal tract of a mammal is controlled. The
pharmaceutical composition includes at least one polyether
ionophore antibiotic or a bicyclic spiroether.
Inventors: |
Berkowitz, Barry;
(Framingham, MA) ; Sachs, George; (Encino, CA)
; Blackburn, Chris; (Natick, MA) |
Correspondence
Address: |
LAHIVE & COCKFIELD
28 STATE STREET
BOSTON
MA
02109
US
|
Assignee: |
Millennium Pharmaceuticals,
Inc.
|
Family ID: |
25421781 |
Appl. No.: |
09/865160 |
Filed: |
May 24, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09865160 |
May 24, 2001 |
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09451622 |
Nov 30, 1999 |
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6271256 |
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09451622 |
Nov 30, 1999 |
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08905985 |
Aug 5, 1997 |
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6017950 |
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Current U.S.
Class: |
514/460 ;
424/653 |
Current CPC
Class: |
Y10S 514/925 20130101;
A61K 31/35 20130101 |
Class at
Publication: |
514/460 ;
424/653 |
International
Class: |
A61K 031/35; A61K
033/24 |
Claims
What is claimed is:
1. A method for controlling gram negative bacteria in a mammal,
comprising administering to a mammal a therapeutically effective
amount of a polyether ionophore antibiotic, such that gram negative
bacteria in the mammal is controlled.
2. The method of claim 1, wherein said mammal is a human.
3. The method of claim 1, wherein said polyether ionophore
antibiotic is selected from the group consisting of monensin,
nigericin and calcimycin and pharmaceutically acceptable salts or
esters thereof.
4. The method of claim 3, wherein said ester is an acetomethoxy
ester.
5. The method of claim 1, wherein said polyether ionophore
antibiotic has the formula 18wherein, R is either phenyl, pyridyl
or aminoethyl-Fmoc and pharmaceutically acceptable salts or esters
thereof.
6. The method of claim 1, wherein said therapeutically effective
amount is between about 1 mg/kg/day and about 100 mg/kg/day.
7. The method of claim 1, wherein said polyether ionophore
antibiotic is administered orally.
8. The method of claim 1, wherein said gram negative bacteria is an
agent associated with disorders of the gastrointestinal tract.
9. The method of claim 8, wherein said gram negative bacteria is
Helicobacter pylori.
10. The method of claim 1, further comprising administering a
proton pump inhibitor, an acid agonist or blocker, a bismuth salt
or combinations thereof in combination with said polyether
ionophore antibiotic.
11. A method for controlling gram negative bacteria in a mammal,
comprising administering to a mammal a therapeutically effective
amount of a bicyclo spiroether compound having at least one
heterocyclic moiety attached to at least one of the ether rings,
such that gram negative bacteria in the mammal is controlled.
12. The method of claim 11, wherein both ether rings are six
membered rings.
13. The method of claim 11, wherein one ether ring is a six
membered ring and one ether ring is five membered.
14. The method of claim 11, wherein at least one heterocyclic
moiety is directly attached to one of the ether rings.
15. The method of claim 14, wherein said heterocyclic moiety is a
tetrahydrofuranyl group.
16. The method of claim 11, wherein at least one heterocyclic
moiety is attached to one of the ether rings via a substituted or
unsubstituted alkylene diradical.
17. The method of claim 16, wherein said heterocyclic moiety is a
tetrahydrofuranyl group.
18. The method of claim 11, wherein said gram negative bacteria is
an agent associated with disorders of the gastrointestinal
tract.
19. The method of claim 18, wherein said gram negative bacteria is
Helicobacter pylori.
20. The method of claim 11, further comprising administering a
proton pump inhibitor, an acid agonist or blocker, a bismuth salt
or combinations thereof in combination with said polyether
ionophore antibiotic.
21. A method for controlling gram negative bacteria in a mammal,
comprising administering to a mammal a therapeutically effective
amount of a compound having the formula: 19wherein, R.sub.1 is a
hydrogen atom or a substituted or unsubstituted alkyl group; A is
either a covalent bond connecting the cyclic ring to B or a
substituted or unsubstituted alkylene diradical connecting the
cyclic ring to B; B is a heterocyclic group; D is a substituted or
unsubstituted alkyl group; the methylene carbon atoms of the cyclic
rings are each independently substituted at one or more positions
with one or two substitutents selected from hydrogen atoms,
hydroxyl groups or substituted or unsubstituted alkyl groups; and
the methine carbon atoms are each independently substituted with
hydrogen atoms or substituted or unsubstituted alkyl groups, such
that gram negative bacteria in the mammal is controlled.
22. The method of claim 21, wherein said heterocyclic group is a
tetrahydrofuranyl group.
23. The method of claim 21, wherein said gram negative bacteria is
an agent associated with disorders of the gastrointestinal
tract.
24. The method of claim 23, wherein said gram negative bacteria is
Helicobacter pylori.
25. The method of claim 21, further comprising administering a
proton pump inhibitor, an acid agonist or blocker, a bismuth salt
or combinations thereof in combination with said polyether
ionophore antibiotic.
26. A method treating a state characterized by the presence of gram
negative bacteria in a mammal, comprising administering to a mammal
a therapeutically effective amount of a polyether ionophore
antibiotic, such that a state characterized by the presence of gram
negative bacteria in a mammal is treated.
27. The method of claim 26, wherein said mammal is a human.
28. The method of claim 26, wherein said polyether ionophore
antibiotic is selected from the group consisting of monensin,
nigericin and calcimycin and pharmaceutically acceptable salts or
esters thereof.
29. The method of claim 28, wherein said ester is an acetomethoxy
ester.
30. The method of claim 26, wherein said polyether ionophore
antibiotic has the formula 20wherein, R is either phenyl, pyridyl
or aminoethyl-Fmoc and pharmaceutically acceptable salts or esters
thereof.
31. The method of claim 26, wherein said therapeutically effective
amount is between about 1 mg/kg/day and about 100 mg/kg/day.
32. The method of claim 26, wherein said polyether ionophore
antibiotic is administered orally.
33. The method of claim 26, wherein said gram negative bacteria is
an agent associated with disorders of the gastrointestinal
tract.
34. The method of claim 33, wherein said gram negative bacteria is
Helicobacter pylori.
35. The method of claim 26, further comprising administering a
proton pump inhibitor, an acid agonist or blocker, a bismuth salt
or combinations thereof in combination with said polyether
ionophore antibiotic.
36. A method for treating a state characterized by the presence of
gram negative bacteria in a mammal, comprising administering to a
mammal a therapeutically effective amount of a bicyclo spiroether
compound having at least one heterocyclic moiety attached to at
least one of the ether rings, such that a state characterized by
the presence of gram negative bacteria in the mammal is
treated.
37. The method of claim 36, wherein both ether rings are six
membered rings.
38. The method of claim 36, wherein one ether ring is a six
membered ring and one ether ring is five membered.
39. The method of claim 36, wherein at least one heterocyclic
moiety is directly attached to one of the ether rings.
40. The method of claim 39, wherein said heterocyclic moiety is a
tetrahydrofuranyl group.
41. The method of claim 36, wherein at least one heterocyclic
moiety is attached to one of the ether rings via a substituted or
unsubstituted alkylene diradical.
42. The method of claim 41, wherein said heterocyclic moiety is a
tetrahydrofuranyl group.
43. The method of claim 36, wherein the gram negative bacteria is
an agent associated with disorders of the gastrointestinal
tract.
44. The method of claim 36, wherein said gram negative bacteria is
Helicobacter pylori.
45. The method of claim 36, further comprising administering a
proton pump inhibitor, an acid agonist or blocker, a bismuth salt
or combinations thereof in combination with said polyether
ionophore antibiotic.
46. A method for treating a state characterized by the presence of
gram negative bacteria in a mammal, comprising administering to a
mammal a therapeutically effective amount of compound having the
formula: 21wherein, R.sub.1 is a hydrogen atom or a substituted or
unsubstituted alkyl group; A is either a covalent bond connecting
the cyclic ring to B or a substituted or unsubstituted alkylene
diradical connecting the cyclic ring to B; B is a heterocyclic
group; D is a substituted or unsubstituted alkyl group; the
methylene carbon atoms of the cyclic rings are each independently
substituted at one or more positions with one or two substitutents
selected from hydrogen atoms, hydroxyl groups or substituted or
unsubstituted alkyl groups; and the methine carbon atoms are each
independently substituted with hydrogen atoms or substituted or
unsubstituted alkyl groups, such that a state characterized by the
presence of gram negative bacteria in a mammal is treated.
47. The method of claim 46, wherein said heterocyclic group is a
tetrahydrofuranyl group.
48. The method of claim 46, wherein the gram negative bacteria is
an agent associated with disorders of the gastrointestinal
tract.
49. The method of claim 48, wherein said gram negative bacteria is
Helicobacter pylori.
50. The method of claim 49, further comprising administering a
proton pump inhibitor, an acid agonist or blocker, a bismuth salt
or combinations thereof in combination with said polyether
ionophore antibiotic.
51. A method for controlling Helicobacter pylori in a mammal,
comprising administering to a mammal a therapeutically effective
amount of a polyether ionophore antibiotic, such that Helicobacter
pylori in the mammal is controlled.
52. The method of claim 51, wherein said polyether ionophore
antibiotic is selected from the group consisting of monensin,
nigericin, calcimycin, 22wherein, R is either phenyl, pyridyl or
aminoethyl-Fmoc and pharmaceutically acceptable salts or esters
thereof.
53. A method for controlling Helicobacter pylori in a mammal,
comprising administering to a mammal a therapeutically effective
amount of a bicyclo spiroether compound having at least one
heterocyclic moiety attached to at least one of the ether rings,
such that Helicobacter pylori in the mammal is controlled.
54. A method for controlling Helicobacter pylori in a mammal,
comprising administering to a mammal a therapeutically effective
amount of a compound having the formula: 23wherein, R.sub.1 is a
hydrogen atom or a substituted or unsubstituted alkyl group; A is
either a covalent bond connecting the cyclic ring to B or a
substituted or unsubstituted alkylene diradical connecting the
cyclic ring to B; B is a heterocyclic group; D is a substituted or
unsubstituted alkyl group; the methylene carbon atoms of the cyclic
rings are each independently substituted at one or more positions
with one or two substitutents selected from hydrogen atoms,
hydroxyl groups or substituted or unsubstituted alkyl groups; and
the methine carbon atoms are each independently substituted with
hydrogen atoms or substituted or unsubstituted alkyl groups, such
that Helicobacter pylori in the mammal is controlled.
55. A method for treating a gastrointestinal disorder in a mammal
characterized by having Helicobacter pylori as a causative agent,
comprising administering to a mammal a therapeutically effective
amount of a polyether ionophore antibiotic, such that a
gastrointestinal disorder characterized by having Helicobacter
pylori as a causative agent is treated.
56. The method of claim 55, wherein said polyether ionophore
antibiotic is selected from the group consisting of monensin,
nigericin, calcimycin, 24wherein, R is either phenyl, pyridyl or
aminoethyl-Fmoc and pharmaceutically acceptable salts or esters
thereof.
57. The method of claim 55, further comprising administering a
proton pump inhibitor, an acid agonist or blocker, a bismuth salt
or combinations thereof in combination with said polyether
ionophore antibiotic.
58. A method for treating a gastrointestinal disorder in a mammal
characterized by having Helicobacter pylori as a causative agent,
comprising administering to a mammal a therapeutically effective
amount of a bicyclo spiroether compound having at least one
heterocyclic moiety attached to at least one of the ether rings,
such that a gastrointestinal disorder characterized by having
Helicobacter pylori as a causative agent is treated.
59. A method for treating a gastrointestinal disorder in a mammal
characterized by having Helicobacter pylori as a causative agent,
comprising administering to a mammal a therapeutically effective
amount of a compound having the formula: 25wherein, R.sub.1 is a
hydrogen atom or a substituted or unsubstituted alkyl group; A is
either a covalent bond connecting the cyclic ring to B or a
substituted or unsubstituted alkylene diradical connecting the
cyclic ring to B; B is a heterocyclic group; D is a substituted or
unsubstituted alkyl group; the methylene carbon atoms of the cyclic
rings are each independently substituted at one or more positions
with one or two substitutents selected from hydrogen atoms,
hydroxyl groups or substituted or unsubstituted alkyl groups; and
the methine carbon atoms are each independently substituted with
hydrogen atoms or substituted or unsubstituted alkyl groups, such
that a gastrointestinal disorder characterized by having
Helicobacter pylori as a causative agent is treated.
60. A method for treating an ulcer in the gastrointestinal tract of
a mammal, comprising administering to a mammal a therapeutically
effective amount of a polyether ionophore antibiotic, such that an
ulcer of the gastrointestinal tract of the mammal is treated.
61. The method of claim 60, wherein said polyether ionophore
antibiotic is selected from the group consisting of monensin,
nigericin, calcimycin, 26wherein, R is either phenyl, pyridyl or
aminoethyl-Fmoc and pharmaceutically acceptable salts or esters
thereof.
62. The method of claim 60, further comprising administering a
proton pump inhibitor, an acid agonist or blocker, a bismuth salt
or combinations thereof in combination with said polyether
ionophore antibiotic.
63. A method for treating an ulcer associated with Helicobacter
pylori in a mammal, comprising administering to a mammal a
therapeutically effective amount of a polyether ionophore
antibiotic, such that an ulcer associated with Helicobacter pylori
in the mammal is treated.
64. The method of claim 63, wherein said polyether ionophore
antibiotic is selected from the group consisting of monensin,
nigericin, calcimycin, 27wherein, R is either phenyl, pyridyl or
aminoethyl-Fmoc and pharmaceutically acceptable salts or esters
thereof.
65. The method of claim 63, further comprising administering a
proton pump inhibitor, an acid agonist or blocker, a bismuth salt
or combinations thereof in combination with said polyether
ionophore antibiotic.
66. A method for treating an ulcer associated with Helicobacter
pylori in a mammal, comprising administering to a mammal a
therapeutically effective amount of a bicyclo spiroether compound
having at least one heterocyclic moiety attached to at least one of
the ether rings, such that an ulcer associated with Helicobacter
pylori in the mammal is treated.
67. A method for treating an ulcer associated with Helicobacter
pylori in a mammal, comprising administering to a mammal a
therapeutically effective amount of a compound having the formula:
28wherein, R.sub.1 is a hydrogen atom or a substituted or
unsubstituted alkyl group; A is either a covalent bond connecting
the cyclic ring to B or a substituted or unsubstituted alkylene
diradical connecting the cyclic ring to B; B is a heterocyclic
group; D is a substituted or unsubstituted alkyl group; the
methylene carbon atoms of the cyclic rings are each independently
substituted at one or more positions with one or two substitutents
selected from hydrogen atoms, hydroxyl groups or substituted or
unsubstituted alkyl groups; and the methine carbon atoms are each
independently substituted with hydrogen atoms or substituted or
unsubstituted alkyl groups, such that an ulcer associated with
Helicobacter pylori in the mammal is treated.
68. A packaged pharmaceutical composition for controlling gram
negative bacteria in a mammal, comprising: a container holding a
therapeutically effective amount of a pharmaceutical composition
for treating gram negative bacteria in a mammal, said
pharmaceutical composition comprising at least one polyether
ionophore antibiotic; and instructions for using said
pharmaceutical composition for control of said gram negative
bacteria, such that gram negative bacteria in the mammal is
controlled.
69. The packaged pharmaceutical of claim 68, wherein said polyether
ionophore antibiotic is selected from the group consisting of
monensin, nigericin and calcimycin and pharmaceutically acceptable
salts or esters thereof.
70. The packaged pharmaceutical of claim 69, wherein said ester is
an acetomethoxy ester thereof.
71. The packaged pharmaceutical of claim 68, wherein said polyether
ionophore antibiotic has the formula 29wherein, R is either phenyl,
pyridyl or aminoethyl-Fmoc.
72. The packaged pharmaceutical of claim 68, wherein said
therapeutically effective amount is between about 1 mg/kg/day and
about 100 mg/kg/day.
73. The packaged pharmaceutical of claim 68, wherein the
pharmaceutical composition is administered orally.
74. The packaged pharmaceutical of claim 68, said package further
comprising a proton pump inhibitor, an acid agonist or blocker, a
bismuth salt or combinations thereof.
Description
BACKGROUND
[0001] Helicobacter pylori bacterial infections are a serious
problem in humans. They have been shown to be a strong causative
factor in gastric ulcer disease, such as stomach ulcers and small
intestine ulcers which can result in death. Helicobacter pylori is
a gram-negative, S-shaped, microaerophilic bacterium that was
discovered and cultured from a human gastric biopsy specimen.
(Warren, J. R. and B. Marshall, (1983) Lancet 1: 1273-1275; and
Marshall et al., (1984) Microbios Lett. 25:83-88). H. pylori
bacterial cells can survive in a low pH environment because of an
enzyme on their outer cell wall called urease. Urease converts urea
in the stomach into bicarbonate and ammonia. The bicarbonate and
ammonia neutralize the acid gastric juices, thereby providing a
protective layer around the H. pylori.
[0002] Since H. pylori are gram-negative rod type bacteria, it is
difficult to treat H. pylori infections without using agents that
will also affect other gram-negative bacteria elsewhere in the
body. H. pylori has been strongly linked to chronic gastritis and
duodenal ulcer disease. (Rathbone et. al., (1986) Gut 27:635-641).
Moreover, evidence is accumulating for an etiologic role of H.
pylori in nonulcer dyspepsia, gastric ulcer disease, and gastric
adenocarcinoma. (Blaser M. J., (1993) Trends Microbiol 1:255-260).
Transmission of the bacteria occurs via the oral route, and the
risk of infection increases with age. (Taylor, D. N. and M. J.
Blaser, (1991) Epidemiol. Rev 13:42-50). H. pylori colonizes the
human gastric mucosa, establishing an infection that usually
persists for decades. Infection by H. pylori is prevalent
worldwide. Developed countries have infection rates over 50% of the
adult population, while developing countries have infection rates
reaching 90% of the adults over the age of 20. (Hopkins R. J. and
J. G. Morris (1994) Am. J. Med 97:265-277).
[0003] The bacterial factors necessary for colonization of the
gastric environment, and for virulence of this pathogen, are poorly
understood. Examples of the putative virulence factors include the
following: urease, an enzyme that may play a role in neutralizing
gastric acid pH (Eaton et al., (1991) Infect. Immunol.
59:2470-2475; Ferrero, R. L. and A. Lee (1991) Microb. Ecol. Hlth.
Dis. 4:121-134; Labigne et al., (1991) J. Bacteriol.
173:1920-1931); the bacterial flagellar proteins responsible for
motility across the mucous layer (Hazell et al., (1986) J. Inf.
Dis. 153:658-663; Leying et al., (1992) Mol. Microbiol.
6:2863-2874; and Haas et al., (1993) Mol. Microbiol. 8:753-760; Vac
A), a bacterial toxin that induces the formation of intracellular
vacuoles in epithelial cells (Schmitt, W. and R. Haas, (1994)
Molecular Microbiol. 12(2: 307-319) and several gastric
tissue-specific adhesions (Boren et al., (1993) Science 262:
1892-1895; Evans et al., (1993) J. Bacteriol. 175:674-683; and Falk
et al., (1993) Proc. Natl. Acad. Sci. USA 90:2035-2039).
[0004] Certain therapeutic agents are known to eradicate H. pylori
infections in vitro. (Huesca et. al., (1993) Zbl. Bakt.
280:244-252; Hopkins, R. J. and J. G. Morris, supra). However, many
agents are suboptimally effective in vivo because of bacterial
resistance, altered drug distribution, patient non-compliance, poor
drug availability and lack of selectivity for H. pylori. (Hopkins,
R. J. and J. G. Morris, supra). Treatment with antibiotics combined
with bismuth are part of a standard regime used to treat H. pylori
infection. (Malfertheiner, P. and J. E. Dominguez-Munoz (1993)
Clinical Therapeutics 15 Supp. B:37-48). Recently, combinations of
a proton pump inhibitor and a single antibiotic have been shown to
ameliorate duodenal ulcer disease. (Malfertheiner, P. and J. E.
Dominguez-Munoz supra).
SUMMARY OF THE INVENTION
[0005] This invention provides methods of controlling gram negative
bacteria and/or treating gram negative infections. In some
embodiments, the methods involve the use of a polyether ionophore
antibiotic to selectively control or kill such bacteria. In a
preferred method, the gram negative bacteria is associated with
gastric Helicobacter pylori infections.
[0006] The present invention provides various methods for
controlling gram negative bacteria, such as Helicobacter pylori, by
administering to a mammal a therapeutically effective amount of a
polyether ionophore antibiotic such that gram negative bacteria in
the mammal is controlled.
[0007] Polyether ionophore antibiotics useful for controlling gram
negative bacteria in a mammal include monensin, nigericin and
calcimycin and pharmaceutically acceptable salts or esters thereof
Preferred polyether antibiotics of the invention include those
having the formula 1
[0008] wherein, R is either phenyl, pyridyl or aminoethyl-Fmoc
(fluorenylmethoxycarbonyl). Coadministration of proton pump
inhibitors, acid agonists or blockers, acid antagonists, bismuth
salts or combinations thereof with polyether ionophore antibiotics
of the invention is particularly preferred.
[0009] The present invention also provides methods for controlling
gram negative bacteria in a mammal by administering to a mammal a
therapeutically effective amount of a bicyclo spiroether compound
having at least one heterocyclic moiety attached to at least one of
the ether rings, such that gram negative bacteria in the mammal is
controlled.
[0010] The present invention provides methods for controlling gram
negative bacteria in a mammal by administering to a mammal a
therapeutically effective amount of a compound having the formula:
2
[0011] such that gram negative bacteria in the mammal is
controlled. R.sub.1 is a hydrogen atom or a substituted or
unsubstituted alkyl group and A is either a covalent bond
connecting the cyclic ring to B or a substituted or unsubstituted
alkylene diradical connecting the cyclic ring to B. B is a
heterocyclic group, preferably a tetrahydrofiranyl group, and D is
a substituted or unsubstituted alkyl group. The methylene carbon
atoms of the cyclic rings are each independently substituted at one
or more positions with one or two substitutents selected from
hydrogen atoms, hydroxyl groups or substituted or unsubstituted
alkyl groups. The methine carbon atoms are each independently
substituted with hydrogen atoms or substituted or unsubstituted
alkyl groups.
[0012] The present invention further provides methods for treating
a state characterized by the presence of gram negative bacteria in
a mammal. In one embodiment, the method includes administering to a
mammal a therapeutically effective amount of a polyether ionophore
antibiotic, such that a state characterized by the presence of gram
negative bacteria in the mammal is treated. In a preferred
embodiment, the gram negative bacteria is Helicobacter pylori.
[0013] In another embodiment, a method includes administering to a
mammal a therapeutically effective amount of a bicyclo spiroether
compound having at least one heterocyclic moiety attached to at
least one of the ether rings, such that a state characterized by
the presence of gram negative bacteria, such as Helicobacter pylori
in the mammal is treated. The polyether ionophore antibiotics and
bicyclo spiroether compounds of the invention are useful for both
prophylactic and/or therapeutic treatments of gram negative
bacteria found in the gastrointestinal tract of a mammal.
[0014] In still another embodiment, a method includes administering
to a mammal a therapeutically effective amount of compound having
the formula: 3
[0015] such that a state characterized by the presence of gram
negative bacteria, such as Helicobacter pylori, in the mammal is
treated. A, B, D and R.sub.1 are as defined supra.
[0016] The present invention provides methods for controlling
Helicobacter pylori in a mammal. In one embodiment, a method
includes administering to a mammal a therapeutically effective
amount of a polyether ionophore antibiotic, such that Helicobacter
pylori in the mammal is controlled.
[0017] In another embodiment, a method includes administering to a
mammal a therapeutically effective amount of a bicyclo spiroether
compound having at least one heterocyclic moiety attached to at
least one of the ether rings, such that Helicobacter pylori in the
mammal is controlled.
[0018] In still another embodiment, a method includes administering
to a mammal a therapeutically effective amount of a compound having
the formula: 4
[0019] wherein A, B, D and R.sub.1 are as defined supra, such that
Helicobacter pylori in the mammal is controlled.
[0020] The present invention further provides methods for treating
a gastrointestinal disorder in a mammal characterized by having
gram negative bacteria, such as Helicobacter pylori, as a causative
agent. In one embodiment, a method includes administration of a
therapeutically effective amount of a polyether ionophore
antibiotic to a mammal, such that a gastrointestinal disorder
characterized by having Helicobacter pylori as a causative agent is
treated.
[0021] In another embodiment, a method includes administering to a
mammal a therapeutically effective amount of a bicyclo spiroether
compound having at least one heterocyclic moiety attached to at
least one of the ether rings, such that a gastrointestinal disorder
characterized by having Helicobacter pylori as a causative agent is
treated.
[0022] In still another embodiment, a method includes administering
to a mammal a therapeutically effective amount of a compound having
the formula: 5
[0023] wherein A, B, D and R.sub.1 are defined supra, such that a
gastrointestinal disorder characterized by having Helicobacter
pylori as a causative agent is treated.
[0024] The present invention also provides methods for treating an
ulcer in the gastrointestinal tract of a mammal. In one embodiment,
the method includes administration of a therapeutically effective
amount of a polyether ionophore antibiotic to a mammal, such that
an ulcer of the gastrointestinal tract of the mammal is
treated.
[0025] The present invention also provides methods for treating an
ulcer associated with Helicobacter pylori in a mammal. In one
embodiment, a method includes administering to a mammal a
therapeutically effective amount of a polyether ionophore
antibiotic, such that an ulcer associated with Helicobacter pylori
in the mammal is treated.
[0026] In another embodiment, a method includes administering to a
mammal a therapeutically effective amount of a bicyclo spiroether
compound having at least one heterocyclic moiety attached to at
least one of the ether rings, such that an ulcer associated with
Helicobacter pylori in the mammal is treated.
[0027] In still another embodiment, the method includes
administering to a mammal a therapeutically effective amount of a
compound having the formula: 6
[0028] wherein A, B, D and R.sub.1 are defined supra, such that an
ulcer associated with Helicobacter pylori in the mammal is
treated.
[0029] The present invention also provides packaged pharmaceutical
compositions for controlling gram negative bacteria. The container
holds a therapeutically effective amount of a pharmaceutical
composition for controlling gram negative bacteria in a mammal and
instructions for using the pharmaceutical composition for control
of the gram negative bacteria. The pharmaceutical composition
includes at least one polyether ionophore antibiotic, such that
growth of gram negative bacteria in a mammal is treated.
DETAILED DESCRIPTION OF THE INVENTION
[0030] The features and other details of the invention will now be
more particularly described and pointed out in the claims. It will
be understood that the particular embodiments of the invention are
shown by way of illustration and not as limitations of the
invention. The principle features of this invention can be employed
in various embodiments without departing from the scope of the
invention.
[0031] The present invention pertains to a method for controlling
gram negative bacteria in a mammal, e.g. control of the growth of
gram negative bacteria. One embodiment includes administering to a
mammal a therapeutically effective amount of a polyether ionophore
antibiotic, such that gram negative bacteria in the mammal is
controlled, e.g., gram negative bacteria associated with disorders
of the gastrointestinal tract of a mammal. Preferred polyether
ionophore antibiotics useful for controlling gram negative bacteria
in a mammal include monensin, nigericin and calcimycin and
pharmaceutically acceptable salts or esters thereof. Particularly
preferred polyether antibiotics of the invention include those
having the formula 7
[0032] wherein, R is either phenyl, pyridyl or aminoethyl-Fmoc
(fluorenylmethoxycarbonyl) and pharmaceutically acceptable salts or
esters thereof. Coadministration of proton pump inhibitors (PPIs),
acid agonists or blockers, bismuth salts or combinations thereof
with polyether ionophore antibiotics of the invention is
particularly preferred.
[0033] In certain embodiments of the invention, the following
polyether ionophore compounds are excluded from use in the method
of the invention: salinomycin, ionomycin, lasalocid and the
benzylated carboxylic acid of monensin.
[0034] The language "controlling gram negative bacteria" is
intended to include changes in growth or replication of the gram
negative bacteria or eradication of gram negative bacteria. The
language includes preventing survival or inhibiting continued
growth and replication of gram negative bacteria In a preferred
embodiment, the control of the gram negative bacteria is such that
a gram negative bacterial infection is treated. In a preferred
embodiment, the control is selective such that a particular target
gram negative bacteria is controlled while other bacteria which is
not detrimental to the mammal is allowed to remain substantially
uncontrolled or substantially unaffected, e.g. gram positive
bacteria.
[0035] The term "gram negative bacteria" is art recognized and is
intended to include those bacteria which, when treated with either
gentian violet, or its analog crystal violet, followed by iodine
and an organic wash do not stain. Typically, a counterstain of some
contrasting color is applied to demonstrate gram negative bacteria.
The preferred gram negative bacteria are those capable of growing
or existing in the gastrointestinal tract of mammals which can be
problematic under some conditions. In one embodiment, preferred
gram negative bacteria is of the genus Helicobacter, more
preferably the species Helicobacter pylori.
[0036] The term "gastrointestinal tract" is art recognized and is
intended to include the digestive system of an animal. The
gastrointestinal tract(s) includes the esophagus, stomach and
intestine, e.g., small and large.
[0037] The language "gram negative bacteria which are agents
associated with disorders of the gastrointestinal tract" is
intended to include those gram negative bacteria which cause or are
associated with disorders of the gastrointestinal tract, including
inflammation and/or ulcers. Disorders of the gastrointestinal tract
also include those symptoms not manifested by the presence of
ulcerations in the gastric mucosa, such as gastritis, non-ulcer
dyspepsia, esophageal reflux disease and peptic ulcer disease.
Helicobacter pylori is an example of a gram negative bacteria which
is associated with disorders of the gastrointestinal tract as
described previously. Additionally, Helicobacter pylori can be
associated with mucosa associated lymphoid tissue (MALT) acne
rosacea, Gulf Veteran's Syndrome, Chronic Fatigue Syndrome and
halitosis. In one embodiment, gram negative bacteria normally found
as flora in the gastrointestinal tract, which do not cause
disorders of the gastrointestinal tract under normal circumstances,
e.g., E. coli., are not encompassed by this terminology.
[0038] The term "mammal," as used herein, refers to an animal, more
preferably a warm-blooded animal, most preferably cattle, sheep,
pigs, horses, dogs, cats, rats, mice, and humans. Mammals
susceptible to infection by gram negative bacteria, e.g.,
Helicobacter pylori, or susceptible to disorders of the
gastrointestinal tract are included as part of this invention.
[0039] The term "therapeutically effective amount" is that amount
necessary or sufficient to control gram negative bacteria in the
above methods, e.g., prevent or reduce the gram negative bacteria's
ability to grow. The term "therapeutically effective amount" also
can be that amount necessary to eradicate, e.g., kill, the gram
negative bacteria. An effective amount of the therapeutic agent
necessary to control gram negative bacteria can vary according to
factors such as the type of gram negative bacteria, the amount of
gram negative bacteria already present in the animal, the age, sex,
and weight of the animal, and the ability of the compounds of the
present invention to control gram negative bacterial in the mammal.
Dependent upon the desired outcome, the amount of control can be
tailored for structure/relationship correlations of the therapeutic
agents.
[0040] A therapeutically effective amount preferably controls the
amount of gram negative bacteria and/or a disease state
characterized by the presence of gram negative bacteria in the
infected mammal by at least about 20%, more preferably by at least
about 40%, even more preferably by at least about 60%, and still
more preferably by at least about 80% relative to untreated
subjects. These percentages refer to a decrease in the amount of
gram negative bacteria found in the infected animal and/or the
decrease in symptoms associated with a disease state characterized
by the presence of gram negative bacteria in the infected animal
relative to untreated subjects. For example, a sample of gastric
juices can be obtained from an animal infected with gram negative
bacteria and treated with one of the compounds described supra. A
second sample of gastric juices can be obtained from an animal also
infected with gram negative bacteria which is left untreated with
any compound. One skilled in the art would be able to develop the
samples on a petri dish (as described in the Examples infra) to
determine whether the compound tested controlled the gram negative
bacteria relative to the untreated sample, e.g., reduction or
elimination of colonies relative to the untreated sample.
Furthermore, the ability of a compound of the invention to control
gram negative bacteria associated with disorders of the
gastrointestinal tract of a mammal can be evaluated in an animal
model system or by standard in vitro assays that may be predictive
of efficacy in treating gram negative bacteria associated with
infection of the gastrointestinal tract in human diseases. Such in
vivo model systems include those described by E. Dick-Hegedus &
A. Lee, Scand. J. Gastroenterol. 26, 909 (1991).
[0041] The term "polyether ionophore antibiotic" is art recognized
and is described in detail in U.S. Pat. Nos. 4,302,450; 3,839,447;
3,719,753; 3,577,531; and 3,873,715, all of which are incorporated
herein by reference. Furthermore, the biology and chemistry of the
polyether ionophore antibiotics are disclosed in Polyether
Antibiotics-Naturally Occurring Acid Ionophores, Volume 1- Biology,
edited by John W. Westley. Westley, Marcel Dekker, Inc. 1982,
ISBNO-9247-1655-8 (v. 1) and Polyether Antibiotics-Naturally
Occurring Acid Ionophores, Volume 2 - Chemistry, Edited by John W.
Westley, Marcel Dekker, Inc., 1983, ISBN D-8247-1888-7 (v. 2).
[0042] One common characteristic of polyether ionophores is that
they are able to complex with monovalent metal ions, such as sodium
and potassium, and render such ions organic solvent soluble. These
polycyclic ethers enfold a metal ion, forming a loop or ball around
the ion with the oxygen function adjacent to the metal and the
hydrocarbon backbone forming the exterior of the complex. This
three dimensional arrangement, coupled with the fact there is no
net charge on the complex, render the whole complex organic solvent
soluble. The molecule is able to maintain its loop or ball
configuration because of hydrogen bonding between the chemical
moieties affixed to opposite ends of the molecule. Usually this
involves a carboxylic acid and a hydroxyl group.
[0043] The "polyether" nature of the ionophore refers to the
considerable number of tetrahydropyrans and tetrahydrofurans found
in the ionophore structure. The compounds are characterized by
cyclic ether moieties in their chemical structures. Examples
include monensin (i.e., A-3823A) 8
[0044] Preferred examples include monensin derivatives 9
[0045] wherein, R is either phenyl, pyridyl or aminoethyl-Fmoc and
pharmaceutically acceptable salts or esters thereof.
[0046] Cyclic polyether ionophore antibiotics, such as nigericin
and monensin, are known to be effective against grain positive type
bacteria infections, but have shown reduced or little effectiveness
against gram negative type bacterial infections. Generally, such
antibiotics have been used primarily in the treatment of non-human
bacterial infections. Their preferred use has been in agrochemical
applications as an additive to fowl feed stocks for the treatment
of coccidiosis (gram-positive) infections of young fowl.
[0047] The term "proton pump inhibitor" is art recognized and
includes those compounds, e.g., benzimidazoles, which inhibit
H.sup.+ and K.sup.+ ATPase of stomach parietal cells which dose
dependently inhibit the urease activity in H. pylori. Suitable
examples of PPIs include lansoprazole
(2-[[[3-methyl4-(2,2,2-trifluoroethoxy)2-pyridyl]methyl]sulf- inyl]
benzimidazole and omeprazole
(5-methoxy-2-[[(4-methoxy-3,5-dimethyl--
2-pyridinyl)methyl]sulfinyl]-1H-benzimidazole.
[0048] The term "acid blocker or agonist" is art recognized and
includes those compounds which slow down the rate of acid secretion
into an individual's stomach. Suitable examples include
Tagamet.RTM. (cimetidine) or Zantac.RTM. (rantidine
hydrochloride).
[0049] The term "acid receptor antagonist" is art recognized and
includes those compounds which inhibit the interaction between a
receptor site and an agent which binds to the site. Suitable
examples include metronidazole (2-methyl-5-nitroimidazole-1-ethanol
and clarithromycin (6-O-methylerythromycin).
[0050] The term "bismuth salt" is art recognized and includes those
compounds which enhance mucus glycoprotein secretion, strengthen
viscoelastic gel properties of mucus, cause increased concentration
of epithelial growth factor in ulcer tissue and stimulate
prostaglandin synthesis in the gastric antral mucosa. A suitable
example of a bismuth compound includes tripotassium dicitrato
bismuth. An over the counter mixture which contains a bismuth
compound is Peptobismal.RTM..
[0051] Proton pump inhibitors, acid blockers and bismuth compounds,
alone or in combination, can be given in conjunction with polyether
ionophore antibiotics or bicyclo spiroethers (described infra) of
the invention.
[0052] As set out above, certain embodiments of the present
compounds can contain a basic functional group, such as amino or
alkylamino, and are, thus, capable of forming pharmaceutically
acceptable salts with pharmaceutically acceptable acids. The term
"pharmaceutically acceptable salts" is art recognized and includes
relatively non-toxic, inorganic and organic acid addition salts of
compounds of the present invention. These salts can be prepared in
situ during the final isolation and purification of the compounds
of the invention, or by separately reacting a purified compound of
the invention in its free base form with a suitable organic or
inorganic acid, and isolating the salt thus formed. Representative
salts include the hydrobromide, hydrochloride, sulfate, bisulfate,
phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate,
laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate,
fuinarate, succinate, tartrate, napthylate, mesylate,
glucoheptonate, lactobionate, and laurylsulphonate salts and the
like. (See, e.g., Berge et al. (1977) "Pharmaceutical Salts", J.
Pharm. Sci. 66:1-19).
[0053] In other cases, the compounds of the present invention may
contain one or more acidic functional groups and, thus, are capable
of forming pharmaceutically acceptable salts with pharmaceutically
acceptable bases. The term "pharmaceutically acceptable salts" in
these instances includes relatively non-toxic, inorganic and
organic base addition salts of compounds of the present invention.
These salts can likewise be prepared in situ during the final
isolation and purification of the compounds, or by separately
reacting the purified compound in its free acid form with a
suitable base, such as the hydroxide, carbonate or bicarbonate of a
pharmaceutically acceptable metal cation, with ammonia, or with a
pharmaceutically acceptable organic primary, secondary or tertiary
amine. Representative alkali or alkaline earth salts include the
lithium, sodium, potassium, calcium, magnesium, and aluminum salts
and the like. Representative organic amines useful for the
formation of base addition salts include ethylamine, diethylamine,
ethylenediamine, ethanolamine, diethanolamine, piperazine and the
like.
[0054] The term "pharmaceutically acceptable esters" refers to the
relatively non-toxic, esterified products of the compounds of the
present invention. Ihese esters can be prepared in situ during the
final isolation and purification of the compounds, or by separately
reacting the purified compound in its free acid form or hydroxyl
with a suitable esterifying agent. Carboxylic acids can be
converted into esters via treatment with an alcohol in the presence
of a catalyst. Hydroxyls can be converted into esters via treatment
with an esterifying agent such as alkanoyl halides. The term is
further intended to include lower hydrocarbon groups capable of
being solvated under physiological conditions, e.g., alkyl esters,
methyl, ethyl and propyl esters. (See, for example, Berge et al.,
supra.) A preferred ester group is an acetomethoxy ester group.
[0055] When the compounds of the present invention are administered
as pharmaceuticals, to humans and mammals, they can be given per se
or as a pharmaceutical composition containing, for example, 0.1 to
99.5% (more preferably, 0.5 to 90%) of active ingredient in
combination with a pharmaceutically acceptable carrier.
[0056] The phrase "pharmaceutically acceptable carrier" is art
recognized and includes a pharmaceutically acceptable material,
composition or vehicle, suitable for administering compounds of the
present invention to mammals. The carriers include liquid or solid
filler, diluent, excipient, solvent or encapsulating material,
involved in carrying or transporting the subject agent from one
organ, or portion of the body, to another organ, or portion of the
body. Each carrier must be "acceptable" in the sense of being
compatible with the other ingredients of the formulation and not
injurious to the patient. Some examples of materials which can
serve as pharmaceutically acceptable carriers include: sugars, such
as lactose, glucose and sucrose; starches, such as corn starch and
potato starch; cellulose, and its derivatives, such as sodium
carboxymethyl cellulose, ethyl cellulose and cellulose acetate;
powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa
butter and suppository waxes; oils, such as peanut oil, cottonseed
oil, safflower oil, sesame oil, olive oil, corn oil and soybean
oil; glycols, such as propylene glycol; polyols, such as glycerin,
sorbitol, mannitol and polyethylene glycol; esters, such as ethyl
oleate and ethyl laurate; agar; buffering agents, such as magnesium
hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water;
isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer
solutions; and other non-toxic compatible substances employed in
pharmaceutical formulations.
[0057] Wetting agents, emulsifiers and lubricants, such as sodium
lauryl sulfate and magnesium stearate, as well as coloring agents,
release agents, coating agents, sweetening, flavoring and perfuming
agents, preservatives and antioxidants can also be present in the
compositions.
[0058] Examples of pharmaceutically acceptable antioxidants
include: water soluble antioxidants, such as ascorbic acid,
cysteine hydrochloride, sodium bisulfate, sodium metabisulfite,
sodium sulfite and the like; oil-soluble antioxidants, such as
ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated
hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol,
and the like; and metal chelating agents, such as citric acid,
ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid,
phosphoric acid, and the like.
[0059] Formulations of the present invention include those suitable
for oral, nasal, topical, transdermal, buccal, sublingual, rectal,
vaginal and/or parenteral administration. The formulations may
conveniently be presented in unit dosage form and may be prepared
by any methods well known in the art of pharmacy. The amount of
active ingredient which can be combined with a carrier material to
produce a single dosage form will generally be that amount of the
compound which produces a therapeutic effect. Generally, out of one
hundred per cent, this amount will range from about 1 per cent to
about ninety-nine percent of active ingredient, preferably from
about 5 per cent to about 70 per cent, most preferably from about
10 per cent to about 30 per cent.
[0060] Methods of preparing these formulations or compositions
include the step of bringing into association a compound of the
present invention with the carrier and, optionally, one or more
accessory ingredients. In general, the formulations are prepared by
uniformly and intimately bringing into association a compound of
the present invention with liquid carriers, or finely divided solid
carriers, or both, and then, if necessary, shaping the product.
[0061] Formulations of the invention suitable for oral
administration may be in the form of capsules, cachets, pills,
tablets, lozenges (using a flavored basis, usually sucrose and
acacia or tragacanth), powders, granules, or as a solution or a
suspension in an aqueous or non-aqueous liquid, or as an
oil-in-water or water-in-oil liquid emulsion, or as an elixir or
syrup, or as pastilles (using an inert base, such as gelatin and
glycerin, or sucrose and acacia) and/or as mouth washes and the
like, each containing a predetermined amount of a compound of the
present invention as an active ingredient. A compound of the
present invention may also be administered as a bolus, electuary or
paste.
[0062] In solid dosage forms of the invention for oral
administration (capsules, tablets, pills, dragees, powders,
granules and the like), the active ingredient is mixed with one or
more pharmaceutically acceptable carriers, such as sodium citrate
or dicalcium phosphate, and/or any of the following: fillers or
extenders, such as starches, lactose, sucrose, glucose, mannitol,
and/or silicic acid; binders, such as, for example,
carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone,
sucrose and/or acacia; humectants, such as glycerol; disintegrating
agents, such as agar-agar, calcium carbonate, potato or tapioca
starch, alginic acid, certain silicates, and sodium carbonate;
solution retarding agents, such as paraffin; absorption
accelerators, such as quaternary ammonium compounds; wetting
agents, such as, for example, cetyl alcohol and glycerol
monostearate; absorbents, such as kaolin and bentonite clay;
lubricants, such a talc, calcium stearate, magnesium stearate,
solid polyethylene glycols, sodium lauryl sulfate, and mixtures
thereof; and coloring agents. In the case of capsules, tablets and
pills, the pharmaceutical compositions may also comprise buffering
agents. Solid compositions of a similar type may also be employed
as fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugars, as well as high molecular
weight polyethylene glycols and the like.
[0063] A tablet may be made by compression or molding, optionally
with one or more accessory ingredients. Compressed tablets may be
prepared using binder (for example, gelatin or hydroxypropylmethyl
cellulose), lubricant, inert diluent, preservative, disintegrant
(for example, sodium starch glycolate or cross-linked sodium
carboxymethyl cellulose), surface-active or dispersing agent.
Molded tablets may be made by molding in a suitable machine a
mixture of the powdered compound moistened with an inert liquid
diluent.
[0064] The tablets, and other solid dosage forms of the
pharmaceutical compositions of the present invention, such as
dragees, capsules, pills and granules, may optionally be scored or
prepared with coatings and shells, such as enteric coatings and
other coatings well known in the pharmaceutical-formulating art.
They may also be formulated so as to provide slow or controlled
release of the active ingredient therein using, for example,
hydroxypropylmethyl cellulose in varying proportions to provide the
desired release profile, other polymer matrices, liposomes and/or
microspheres. They may be sterilized by, for example, filtration
through a bacteria-retaining filter, or by incorporating
sterilizing agents in the form of sterile solid compositions which
can be dissolved in sterile water, or some other sterile injectable
medium immediately before use. These compositions may also
optionally contain opacifying agents and may be of a composition
that they release the active ingredient(s) only, or preferentially,
in a certain portion of the gastrointestinal tract, optionally, in
a delayed manner. Examples of embedding compositions which can be
used include polymeric substances and waxes. The active ingredient
can also be in micro-encapsulated form, if appropriate, with one or
more of the above-described excipients.
[0065] Liquid dosage forms for oral administration of the compounds
of the invention include pharmaceutically acceptable emulsions,
microemulsions, solutions, suspensions, syrups and elixirs. In
addition to the active ingredient, the liquid dosage forms may
contain inert diluents commonly used in the art, such as, for
example, water or other solvents, solubilizing agents and
emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl
carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, oils (in particular,
cottonseed, groundnut, corn, germ, olive, castor and sesame oils),
glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty
acid esters of sorbitan, and mixtures thereof.
[0066] Besides inert diluents, the oral compositions can also
include adjuvants such as wetting agents, emulsifying and
suspending agents, sweetening, flavoring, coloring, perfuming and
preservative agents.
[0067] Suspensions, in addition to the active compounds, may
contain suspending agents as, for example, ethoxylated isostearyl
alcohols, polyoxyethylene sorbitol and sorbitan esters,
microcrystalline cellulose, aluminum metahydroxide, bentonite,
agar-agar and tragacanth, and mixtures thereof.
[0068] Formulations of the pharmaceutical compositions of the
invention for rectal or vaginal administration may be presented as
a suppository, which may be prepared by mixing one or more
compounds of the invention with one or more suitable nonirritating
excipients or carriers comprising, for example, cocoa butter,
polyethylene glycol, a suppository wax or a salicylate, and which
is solid at room temperature, but liquid at body temperature and,
therefore, will melt in the rectum or vaginal cavity and release
the active compound.
[0069] Formulations of the present invention which are suitable for
vaginal administration also include pessaries, tampons, creams,
gels, pastes, foams or spray formulations containing such carriers
as are known in the art to be appropriate.
[0070] Dosage forms for the topical or transdermal administration
of a compound of this invention include powders, sprays, ointments,
pastes, creams, lotions, gels, solutions, patches and inhalants.
The active compound may be mixed under sterile conditions with a
pharmaceutically acceptable carrier, and with any preservatives,
buffers, or propellants which may be required.
[0071] The ointments, pastes, creams and gels may contain, in
addition to an active compound of this invention, excipients, such
as animal and vegetable fats, oils, waxes, paraffins, starch,
tragacanth, cellulose derivatives, polyethylene glycols, silicones,
bentonites, silicic acid, talc and zinc oxide, or mixtures
thereof.
[0072] Powders and sprays can contain, in addition to a compound of
this invention, excipients such as lactose, talc, silicic acid,
aluminum hydroxide, calcium silicates and polyamide powder, or m of
these substances. Sprays can additionally contain customary
propellants, such as chlorofluorohydrocarbons and volatile
unsubstituted hydrocarbons, such as butane and propane.
[0073] Transdermal patches have the added advantage of providing
controlled delivery of a compound of the present invention to the
body. Such dosage forms can be made by dissolving or dispersing the
compound in the proper medium. Absorption enhancers can also be
used to increase the flux of the compound across the skin. The rate
of such flux can be controlled by either providing a rate
controlling membrane or dispersing the active compound in a polymer
matrix or gel.
[0074] Ophthalmic formulations, eye ointments, powders, solutions
and the like, are also contemplated as being within the scope of
this invention.
[0075] Pharmaceutical compositions of this invention suitable for
parenteral administration comprise one or more compounds of the
invention in combination with one or more pharmaceutically
acceptable sterile isotonic aqueous or nonaqueous solutions,
dispersions, suspensions or emulsions, or sterile powders which may
be reconstituted into sterile injectable solutions or dispersions
just prior to use, which may contain antioxidants, buffers,
bacteriostats, solutes which render the formulation isotonic with
the blood of the intended recipient or suspending or thickening
agents.
[0076] Examples of suitable aqueous and nonaqueous carriers which
may be employed in the pharmaceutical compositions of the invention
include water, ethanol, polyols (such as glycerol, propylene
glycol, polyethylene glycol, and the like), and suitable mixtures
thereof, vegetable oils, such as olive oil, and injectable organic
esters, such as ethyl oleate. Proper fluidity can be maintained,
for example, by the use of coating materials, such as lecithin, by
the maintenance of the required particle size in the case of
dispersions, and by the use of surfactants.
[0077] These compositions may also contain adjuvants such as
preservatives, wetting agents, emulsifying agents and dispersing
agents. Prevention of the action of microorganisms may be ensured
by the inclusion of various antibacterial and antifungal agents,
for example, paraben, chlorobutanol, phenol sorbic acid, and the
like. It may also be desirable to include isotonic agents, such as
sugars, sodium chloride, and the like into the compositions. In
addition, prolonged absorption of the injectable pharmaceutical
form may be brought about by the inclusion of agents which delay
absorption such as aluminum monostearate and gelatin.
[0078] In some cases, in order to prolong the effect of a drug, it
is desirable to slow the absorption of the drug from subcutaneous
or intramuscular injection. This may be accomplished by the use of
a liquid suspension of crystalline or amorphous material having
poor water solubility. The rate of absorption of the drug then
depends upon its rate of dissolution which, in turn, may depend
upon crystal size and crystalline form. Alternatively, delayed
absorption of a parenterally-administered drug form is accomplished
by dissolving or suspending the drug in an oil vehicle.
[0079] Injectable depot forms are made by forming microencapsule
matrices of the subject compounds in biodegradable polymers such as
polylactide-polyglycolide. Depending on the ratio of drug to
polymer, and the nature of the particular polymer employed, the
rate of drug release can be controlled. Examples of other
biodegradable polymers include poly(orthoesters) and
poly(anhydrides). Depot injectable formulations are also prepared
by entrapping the drug in liposomes or microemulsions which are
compatible with body tissue.
[0080] The preparations of the present invention may be given
orally, parenterally, topically, or rectally. They are of course
given by forms suitable for each administration route. For example,
they are administered in tablets or capsule form, by injection,
inhalation, eye lotion, ointment, suppository, etc. administration
by injection, infusion or inhalation; topical by lotion or
ointment; and rectal by suppositories. Oral administration is
preferred.
[0081] The phrases "parenteral administration" and "administered
parenterally" as used herein means modes of administration other
than enteral and topical administration, usually by injection, and
includes, without limitation, intravenous, intramuscular,
intrarterial, intrathecal, intracapsular, intraorbital,
intracardiac, intradermal, intraperitoneal, transtracheal,
subcutaneous, subcuticular, intraarticular, subcapsular,
subarachnoid, intraspinal and intrasternal injection and
infusion.
[0082] The phrases "systemic administration," "administered
systemically," "peripheral administration" and "administered
peripherally" as used herein mean the administration of a compound,
drug or other material other than directly into the central nervous
system, such that it enters the patient's system and, thus, is
subject to metabolism and other like processes, for example,
subcutaneous administration.
[0083] These compounds may be administered to humans and other
animals for therapy by any suitable route of administration,
including orally, nasally, as by, for example, a spray, rectally,
intravaginally, parenterally, intracisternally and topically, as by
powders, ointments or drops, including buccally and
sublingually.
[0084] Regardless of the route of administration selected, the
compounds of the present invention, which may be used in a suitable
hydrated form, and/or the pharmaceutical compositions of the
present invention, are formulated into pharmaceutically acceptable
dosage forms by conventional methods known to those of skill in the
art.
[0085] Actual dosage levels of the active ingredients in the
pharmaceutical compositions of this invention may be varied so as
to obtain an amount of the active ingredient which is effective to
achieve the desired therapeutic response for a particular patient,
composition, and mode of administration, without being toxic to the
patient
[0086] The selected dosage level will depend upon a variety of
factors including the activity of the particular compound of the
present invention employed, or the ester, salt or amide thereof,
the route of administration, the time of administration, the rate
of excretion of the particular compound being employed, the
duration of the treatment, other drugs, compounds and/or materials
used in combination with the particular compound employed, the age,
sex, weight, condition, general health and prior medical history of
the patient being treated, and like factors well known in the
medical arts.
[0087] A physician or veterinarian having ordinary skill in the art
can readily determine and prescribe the effective amount of the
pharmaceutical composition required. For example, the physician or
veterinarian could start doses of the compounds of the invention
employed in the pharmaceutical composition at levels lower than
that required in order to achieve the desired therapeutic effect
and gradually increase the dosage until the desired effect is
achieved.
[0088] In general, a suitable daily dose of a compound of the
invention will be that amount of the compound which is the lowest
dose effective to produce a therapeutic effect. Such an effective
dose will generally depend upon the factors described above.
Generally, intravenous and subcutaneous doses of the compounds of
this invention for a patient, when used for the indicated analgesic
effects, will range from about 0.0001 to about 100 mg per kilogram
of body weight per day, more preferably from about 0.01 to about 50
mg per kg per day, and still more preferably from about 1.0 to
about 100 mg per kg per day. An effective amount is that amount
which controls gram negative bacteria and/or treats a disease state
characterized by the presence of a gram negative bacteria, such as
Helicobacter pylori.
[0089] If desired, the effective daily dose of the active compound
may be administered as two, three, four, five, six or more
sub-doses administered separately at appropriate intervals
throughout the day, optionally, in unit dosage forms.
[0090] While it is possible for a compound of the present invention
to be administered alone, it is preferable to administer the
compound as a pharmaceutical composition.
[0091] The invention also pertains to a second method for
controlling gram negative bacteria in a mammal, e.g., those gram
negative bacteria which are agents associated with disorders of the
gastrointestinal tract, using a bicyclo spiroether compound. The
method includes administering to a mammal a therapeutically
effective amount of a bicyclo spiroether compound having at least
one heterocyclic moiety attached to at least one of the ether rings
to a mammal, such that gram negative bacteria in the mammal is
controlled. Preferably, a tetrahydrofuranyl group is attached to a
ether ring via a substituted or unsubstituted alkylene
diradical.
[0092] The term "bicyclo spiroether" is art recognized and is
intended to include those ethers having one carbon atom common to
both rings. Cyclic ethers useful as a first ring of the bicyclic
structure of bicyclo spiroethers include those ethers having a
carbon backbone chain of three to twenty carbon atoms, preferably
four to seven carbon atoms. Cyclic ethers useful as the second ring
of the bicyclo spiroethers includes those having a carbon backbone
(methylenes) of three to twenty carbon atoms, preferably four to
seven carbon atoms. In a preferred embodiment, both the first and
second ring of the bicyclo spiroether are six membered rings (e.g.,
five carbon atoms in each ring). In another preferred embodiment, a
first ring of the bicyclo spiroether is a six membered ring (e.g.,
five carbon atoms) and the second ring of the bicyclo spiroether is
a five membered ring (e.g., four carbon atoms).
[0093] It is to be understood that the methylene carbon atoms of
both the first and second cyclic ether rings can be each
independently substituted at one or more positions with one or two
substitutents selected from hydrogen atoms, hydroxyl groups or
substituted or unsubstituted alkyl groups.
[0094] The term "heterocyclic moiety" is intended to include 3- to
10-membered ring structures, more preferably 4- to 7-membered
rings, which ring structures include one to four heteroatoms.
Heterocyclic groups include pyrrolidine, oxolane, thiolane,
oxazole, piperidine, piperazine, morpholine, lactones, lactams such
as azetidinones and pyrrolidinones, laciones, sultams, sultones,
and the like. The heterocyclic ring can be substituted at one or
more positions with such substituents as described above, as for
example, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,
alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkyl amino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, sulfonato, sulfamoyl,
sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,
alkylaryl, or an aromatic or heteroaromatic moiety. A heteroalkyl
moiety is an alkyl substituted with a heteroaromatic group.
[0095] The term "heteroatom" as used herein means an atom of any
element other than carbon or hydrogen. Preferred heteroatoms are
nitrogen, oxygen, sulfur and phosphorus.
[0096] The term "tetrahyrofuranyl" group is art recognized and is
intended to include substituted and unsubstituted tetrahydrofuran
groups of the formulae: 10
[0097] The term "alkylene diradical" is intended to include an
organic moiety, such as an aliphatic or aromatic group, which is
attached at two positions to two separate organic moieties.
Suitable non-limiting examples of alkylene diradicals include
11
[0098] The alkylene diradical can be substituted or unsubstituted
with those groups described infra for alkyl and aryl groups.
[0099] The invention further pertains to a method for controlling
gram negative bacteria in a mammal, e.g., such as gram negative
bacteria which are agents associated with disorders of the
gastrointestinal tract including compounds having formulae
described below. The method includes administering to a mammal a
therapeutically effective amount of a compound having the formula:
12
[0100] such that gram negative bacteria in the mammal is
controlled. R.sub.1 is a hydrogen atom or a substituted or
unsubstituted alkyl group and A is either a covalent bond
connecting the cyclic ring to B or a substituted or unsubstituted
alkylene diradical connecting the cyclic ring to B. B is a
heterocyclic group, preferably a tetrahydrofuryl group, and D is a
substituted or unsubstituted alkyl group. The methylene carbon
atoms of the cyclic rings are each independently substituted at one
or more positions with one or two substitutents selected from
hydrogen atoms, hydroxyl groups or substituted or unsubstituted
alkyl groups. The methine carbon atoms are each independently
substituted with hydrogen atoms or substituted or unsubstituted
alkyl groups.
[0101] The term "alkyl" refers to the radical of saturated
aliphatic groups, including straight-chain alkyl groups,
branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl
substituted cycloalkyl groups, and cycloalkyl substituted alkyl
groups. In preferred embodiments, a straight chain or branched
chain alkyl has 30 or fewer carbon atoms in its backbone (e.g.,
C.sub.1-C.sub.30 for straight chain, C.sub.3-C.sub.30 for branched
chain), and more preferably 20 or fewer. Likewise, preferred
cycloalkyls have from 4-10 carbon atoms in their ring structure,
and more preferably have 5, 6 or 7 carbons in the ring
structure.
[0102] Moreover, the term alkyl as used throughout the
specification and claims is intended to include both "unsubstituted
alkyls" and "substituted alkyls", the latter of which refers to
alkyl moieties having substituents replacing a hydrogen on one or
more carbons of the hydrocarbon backbone. Such substituents can
include, for example, halogen, hydroxyl, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl,
alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato,
cyano, amino (including alkyl amino, dialkylamino, arylamino,
diarylamino, and alkylarylamino), acylamino (including
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),
amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,
sulfates, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an
aromatic or heteroaromatic moiety. It will be understood by those
skilled in the art that the moieties substituted on the hydrocarbon
chain can themselves be substituted, if appropriate. Cycloalkyls
can be further substituted, e.g., with the substituents described
above. An "alkylaryl" moiety is an alkyl substituted with an aryl
(e.g., phenylmethyl (benzyl)).
[0103] The term "aryl" as used herein, refers to the radical of
aryl groups, including 5- and 6-membered single-ring aromatic
groups that may include from zero to four heteroatoms, for example,
benzene, pyrrole, furan, thiophene, imidazole, benzoxazole,
benzothiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine
and pyrimidine, and the like. Aryl groups also include polycyclic
fused aromatic groups such as naphthyl, quinolyl, indolyl, and the
like. Those aryl groups having heteroatoms in the ring structure
may also be referred to as "aryl heterocycles", "heteroaryls" or
"heteroaromatics". The aromatic ring can be substituted at one or
more ring positions with such substituents as described above, as
for example, halogen, hydroxyl, alkoxy, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl,
alkylthiocarbonyl, phosphate, phosphonato, phosphinato, cyano,
amino (including alkyl amino, dialkylamino, arylamino, diarylamino,
and alkylarylamino), acylamino (including alkylcarbonylamino,
arylcarbonylamino, carbamoyl and ureido), amidino, imino,
sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,
sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano,
azido, heterocyclyl, alkylaryl, or an aromatic or heteroaromatic
moiety. Aryl groups can also be fused or bridged with alicyclic or
heterocyclic rings which are not aromatic so as to form a polycycle
(e.g., tetralin).
[0104] The term "alkenyl" refers to unsaturated aliphatic groups
analogous in length and possible substitution to the alkyl groups
described above, but that contain at least one double bond.
[0105] The terms "alkoxyalkyl" and "thioalkoxyalkyl" refer to alkyl
groups, as described above, which further include oxygen or sulfur
atoms replacing one or more carbons of the hydrocarbon backbone,
e.g., catenary oxygen or sulfur atoms.
[0106] Unless the number of carbons is otherwise specified, "lower
alkyl" as used herein means an alkyl group, as defined above, but
having from one to ten carbons, more preferably from one to six
carbon atoms in its backbone structure. Preferred alkyl groups are
lower alkyls having one to three carbon atoms.
[0107] The terms "polycyclyl" or "polycyclic radical" refer to the
radical of two or more cyclic rings (e.g., cycloalkyls,
cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls) in which
two or more carbons are common to two adjoining rings, e.g., the
rings are "fused rings". Rings that are joined through non-adjacent
atoms are termed "bridged" rings. Each of the rings of the
polycycle can be substituted with such substituents as described
above, as for example, halogen, hydroxyl, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl,
alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato,
cyano, amino (including alkyl amino, dialkylamino, arylamino,
diarylamino, and alkylarylamino), acylamino (including
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),
amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,
sulfates, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkyl, alkylaryl, or
an aromatic or heteroaromatic moiety.
[0108] The term "heteroatom" as used herein means an atom of any
element other than carbon or hydrogen. Preferred heteroatoms are
nitrogen, oxygen, sulfur and phosphorus.
[0109] It will be noted that the structure of some of the compounds
of this invention includes asymmetric carbon atoms. It is to be
understood accordingly that the isomers arising from such asymmetry
(e.g., all enantiomers and diastereomers) are included within the
scope of this invention, unless indicated otherwise. Such isomers
can be obtained in substantially pure form by classical separation
techniques and by stereochemically controlled synthesis.
[0110] The term "methine" carbon is art recognized and is intended
to include those carbons having a single site for substitution,
e.g., by a hydrogen atom or substituted or unsubstituted alkyl
groups.
[0111] The present invention further pertains to methods for
treating a state, e.g., disorder or condition, characterized by the
presence of gram negative bacteria in a mammal. An example of such
a gram negative bacteria is Helicobacter pylori, which is an agent
associated with disorders of the gastrointestinal tract of mammals
In one embodiment, a method includes administering to a mammal a
therapeutically effective amount of a polyether ionophore
antibiotic, such that a state characterized by the presence of gram
negative bacteria in the mammal is treated.
[0112] In another embodiment, a method includes administering to a
mammal a therapeutically effective amount of a bicyclo spiroether
compound having at least one heterocyclic moiety attached to at
least one of the ether rings, such that a state characterized by
the presence of gram negative bacteria, such as Helicobacter pylori
in the mammal is treated. The polyether ionophore antibiotics and
bicyclo spiroether compounds of the invention are useful for both
prophylactic and/or therapeutic treatments of gram negative
bacteria found in the gastrointestinal tract of a mammal.
[0113] In still another embodiment, a method includes administering
to a mammal a therapeutically effective amount of compound having
the formula: 13
[0114] such that a state characterized by the presence of gram
negative bacteria, such as Helicobacter pylori, in the mammal is
treated. A, B, D and R.sub.1 are as defined supra.
[0115] The language "state characterized by the presence of gram
negative bacteria" is intended to include those diseases, disorders
or conditions which have been associated with a gram negative
bacteria in that the gram negative bacteria is directly or
indirectly a causative agent of the disease, disorder or condition.
The gram negative bacteria does not have to be the sole causative
agent of the disease, disorder or condition merely responsible for
causing some of the symptoms typically associated with the disease,
disorder, or condition being treated. The gram negative bacteria
can be the causative agent alone or at least one other agent can be
involved in the state being treated. Examples include ulcers and
inflammation and those symptoms not manifested by the presence of
ulcerations in the gastric mucosa, such as gastritis, non-ulcer
dyspepsia, esophageal reflux disease and peptic ulcer disease;
typhus (Salmonella typhi), food poisoning (E. coli O157:H7),
bascillary dysentery (Shigella dysenteria), pneumonia ((Psuedomonas
aerugenosa) and cholera (Vivrio cholerae). Preferred examples
include those symptoms associated with Helicobacter pylori.
[0116] The language "treating or treatment of the state
characterized by the presence of gram negative bacteria" is
intended to include the alleviation of or diminishment of at least
one symptom typically associated with the state. The treatment also
includes alleviation or diminishment of more than one symptom.
Preferably, the treatment cures, e.g., substantially eliminates,
the symptoms associated with the state.
[0117] The language "therapeutically effective amount" of a
polyether ionophore, a bicyclo spiroether compound, or those
compounds described supra, is that amount necessary or sufficient
to perform its intended function within a mammal, e.g., eradicate
or inhibit growth of the gram negative bacteria and/or treat the
targeted state. The therapeutically effective amount can vary
depending on such factors as the type of gram negative bacteria,
the weight of the mammal, or the severity of the symptom(s). One of
ordinary skill in the art would be able to study the aforementioned
factors and make a determination regarding the effective amount of
the compound required without undue experimentation. An in vitro or
in vivo assay also can be used to determine an "effective amount"
of the compounds described supra. The ordinarily skilled artisan
would select an appropriate amount of the compound for use in the
aforementioned assay.
[0118] The present invention also pertains to methods for
controlling Helicobacter pylori in a mammal. In one embodiment, a
method includes administering to a mammal a therapeutically
effective amount of a polyether ionophore antibiotic, such that
Helicobacter pylori in the mammal is controlled.
[0119] In another embodiment, a method includes administering to a
mammal a therapeutically effective amount of a bicyclo spiroether
compound having at least one heterocyclic moiety attached to at
least one of the ether rings, such that Helicobacter pylori in the
mammal is controlled.
[0120] In still another embodiment, a method includes administering
to a mammal a therapeutically effective amount of a compound having
the formula: 14
[0121] wherein A, B, D and R.sub.1 are as defined supra, such that
Helicobacter pylori in the mammal is controlled.
[0122] The present invention further provides methods for treating
a gastrointestinal disorder in a mammal characterized by having
gram negative bacteria, such as Helicobacter pylori, as a causative
agent. In one embodiment, a method includes administration of a
therapeutically effective amount of a polyether ionophore
antibiotic to a mammal, such that a gastrointestinal disorder
characterized by having Helicobacter pylori as a causative agent is
treated.
[0123] In another embodiment, a method includes administering to a
mammal a therapeutically effective amount of a bicyclo spiroether
compound having at least one heterocyclic moiety attached to at
least one of the ether rings, such that a gastrointestinal disorder
characterized by having Helicobacter pylori as a causative agent is
treated.
[0124] In still another embodiment, a method includes administering
to a mammal a therapeutically effective amount of a compound having
the formula: 15
[0125] wherein A, B, D and R.sub.1 are defined supra, such that a
gastrointestinal disorder characterized by having Helicobacter
pylori as a causative agent is treated.
[0126] The phrase "gastrointestinal disorder" is intended to
include physical manifestations caused by a foreign stimulus or by
gram negative bacteria, e.g., Helicobacter pylori in the
gastrointestinal tract. Gastrointestinal disorders and can take the
form of inflammation or ulceration of a portion of the
gastrointestinal tract or gastritis, non-ulcer dyspepsia,
esophageal reflux disease and peptic ulcer disease. Preferred
examples include those manifestations caused by or associated with
Helicobacter pylori.
[0127] The languge "treating a gastrointestinal disorder" is
intended to include the alleviation of or diminishment of at least
one symptom typically associated with the disorder. The treatment
also includes alleviation or diminishment of more than one
disorder. Preferably, the treatment cures, e.g., substantially
eliminates, the symptoms associated with the disorder, e.g.
inflammation, ulceration, gastritis, etc..
[0128] The language "therapeutically effective amount" of a
polyether ionophore, a bicyclo spiroether compound, or those
compounds described supra, is that amount necessary or sufficient
to perform its intended function within a mammal, e.g., reduce,
inhibit or eliminate symptoms associated with a gastrointestinal
disorder as a result of the presence of a gram negative bacteria,
such as Helicobacter pylori, in the mammal. The therapeutically
effective amount can vary depending on such factors as the amount
of gram negative bacteria present, the weight of the mammal, or the
severity of the disorder(s). One of ordinary skill in the art would
be able to study the aforementioned factors and make a
determination regarding the effective amount of the compound
required without undue experimentation.
[0129] The present invention also provides methods for treating an
ulcer in the gastrointestinal tract of a mammal. In one embodiment,
the method includes administration of a therapeutically effective
amount of a polyether ionophore antibiotic to a mammal, such that
an ulcer of the gastrointestinal tract of the mammal is
treated.
[0130] The present invention also provides methods for treating an
ulcer associated with Helicobacter pylori in a mammal. In one
embodiment, a method includes administering to a mammal a
therapeutically effective amount of a polyether ionophore
antibiotic, such that an ulcer associated with Helicobacter pylori
in the mammal is treated.
[0131] In another embodiment, a method includes administering to a
mammal a therapeutically effective amount of a bicyclo spiroether
compound having at least one heterocyclic moiety attached to at
least one of the ether rings, such that an ulcer associated with
Helicobacter pylori in the mammal is treated.
[0132] In still another embodiment, the method includes
administering to a mammal a therapeutically effective amount of a
compound having the formula: 16
[0133] wherein A, B, D and R.sub.1 are defined supra, such that an
ulcer associated with Helicobacter pylori in the mammal is
treated
[0134] The term "ulcer" is art recognized and is intended to breaks
in the skin or mucous membrane with loss of surface tissue, e.g.
degradation of the lining of the esophagus, stomach lining, and/or
lining of the small and/or large intestine. Ulcers, such as peptic
ulcers, can be caused by gram negative bacteria such as
Helicobacter pylori.
[0135] The language "an ulcer associated with Helicobacter pylori"
is intended to include ulcers, inflammation, and/or erosion of the
gastrointestinal lining which causes discomfort to the mammal and
is associated directly or indirectly with the presence of
Helicobacter pylori in the mammal.
[0136] The language "treating an ulcer associated with Helicobacter
pylori" is intended to include the alleviation of or diminishment
of at least one symptom typically associated with an ulcer
associated with Helicobacter pylori. The treatment also includes
alleviation or diminishment of more than one symptom. Preferably,
the treatment cures, e.g., substantially eliminates, the symptoms
associated with the ulcer.
[0137] The language "therapeutically effective amount" of a
polyether ionophore, a bicyclo spiroether compound, or those
compounds described supra, is that amount necessary or sufficient
to perform its intended function within a mammal, e.g., reduce,
inhibit or eliminate symptoms associated with an ulcer as a result
of the presence of Helicobacter pylori in the mammal. The
therapeutically effective amount can vary depending on such factors
as the amount of Helicobacter pylori present, the weight of the
mammal, or the severity of the symptom(s). One of ordinary skill in
the art would be able to study the aforementioned factors and make
a determination regarding the effective amount of the compound
required without undue experimentation.
[0138] The present invention pertains to a packaged pharmaceutical
composition for controlling gram negative bacteria. The container
holds a therapeutically effective amount of a pharmaceutical
composition for controlling gram negative bacteria and instructions
for using the pharmaceutical composition for control of the gram
negative bacteria. The pharmaceutical composition includes at least
one polyether ionophore antibiotic, such that gram negative
bacteria is controlled. Polyether ionophore antibiotics include
monensin, nigericin and calcimycin and pharmaceutically acceptable
salts or esters thereof. Preferred polyether ionophore antibiotics
include compounds 17
[0139] having the formula wherein, R is either phenyl, pyridyl or
aminoethyl-Fmoc and pharmaceutically acceptable salts and esters
thereof.
[0140] The invention is further illustrated by the following
examples which in no way should be construed as being further
limiting. The contents of all references, pending patent
applications and published patent applications, cited throughout
this application are hereby incorporated by reference. It should be
understood that the in vitro assays and/or correlated animal models
used throughout the examples are accepted models and that the
demonstration of efficacy in these models is predictive of efficacy
in humans.
EXEMPLIFICATION
Synthesis of Monensin Derivatives
[0141] Synthesis of monensin benzyl ester
[0142] Monensin sodium salt (1.0 g, 1.4 mmol) in acetonitrile (20
ml) was treated with diazabicyclo [2.2.2] octane (0.3 g, 2.7 mmol)
followed by benzyl bromide (2 g, 11.7 mmol) and crypt and [2.2.2].
The reaction mixture was stirred at room temperature with exclusion
of light for 14 days. After 10 days an additional quantity of
benzyl bromide (2 g, 11.7 mmol) was added. The solvent was
evaporated and the crude product redissolved in dichloromethane,
washed with deionized water and dried over magnesium sulfate. The
crude product was chromatographed on silica gel eluting with
hexane-ethyl acetate gradient. Excess benzyl bromide eluted from
the column first followed by benzylmonensin which was isolated as a
colorless oil after removal of the solvent (280 mg, 26%), (MS,
positive ion ESI showed 783 peak; calc. for sodiated product 783).
This compound has been reported previously (Tsukube J. Org. Chem.
56, 875, 1991).
[0143] Synthesis of 26-O-arylcarbamoylmonensins
[0144] These compounds were prepared using a modification of the
procedure described by Westley and co-workers for the synthesis of
the 26-O-carbamoylmonensins. (Westley, et al., Antibiotics 34,
1195, 1983).
[0145] 26-O Phenylmonensin
[0146] A solution of monensin (400 mg, 0.6 mmol) in dry toluene (2
ml) was treated with phenylisocyanate (71 mg, 0.6 mmol) and the
reaction mixture stirred at ambient temperature with exclusion of
light for 24 h. The toluene was evaporated under reduced pressure
and crude product purified by column chromatography on silica gel
eluting with hexane-ethyl acetate gradient. A major UV-active band,
which had a higher R.sub.1 value than residual monensin, was
evaporated to give a white crystalline solid 130 mg, 27% (MS,
positive ion ESI showed 811 peak; calc mass for sodiated adduct of
product 811).
[0147] 26-O-(3-pyridylcarbamoyl) monensin
[0148] 3-pyridylcarbamoyl monensin compound was prepared as above
in 20% yield by reaction of monensin with 3-pyridylisocyanate (A.
Orahovats Helv. Chimica Acata 70, 262, 1987) followed by
chromatography as described above. (MS, negative ion ESI showed 789
peak; calc mass 790).
[0149] 26-O-[1-(Fmoc-amino) ethylcarbamoyl] monensin
[0150] Fmoc-amino ethylcarbamoyl monensin was prepared as above in
35% yield by reaction of monensin with 1-(Fmoc-amino)
ethylisocyanate followed by column chromatography on silica eluting
with ethyl acetate and 5% methanol in ethylacetate. (MS, negative
ion ESI showed 977 peak; calc mass 978).
Minimum Inhibitory Concentration (MIC) of Monensin and Nigericin
against H. pylori
[0151] Drug Preparation
[0152] Monensin (Sigma Cat. No. M-5273) was dissolved in 100%
absolute ethanol (AAPER Alcohol and Chemical Co.) at a
concentration of 20 mg/ml. Subsequent dilutions were made in
ethanol immediately prior to use. Nigericin (Sigma Cat. No. N-7143)
was dissolved in DMSO at 2 mg/ml. Subsequent dilutions were made in
BHI medium (Brain heart infusion broth, Difco #0037-07-0)
immediately prior to use.
1 Initial Concentration Final Concentrations (1:200) 2 mg/ml 10
.mu.g/ml 200 .mu.g/ml 1 .mu.g/ml 20 .mu.g/ml 0.1 .mu.g/ml 2
.mu.g/ml 10 ng/ml 200 ng/ml 1 ng/ml 20 ng/ml 0.1 ng/ml
[0153] Media Preparation
[0154] Blood agar base (BAB) was prepared containing 1.5% agar and
autoclaved. 20 ml samples of autoclaved BAB were poured into 50 mL
tubes and placed in a 56.degree. C. water bath.
[0155] Plate Preparation
[0156] 100 .mu.l of a drug (final concentration equivalent to 1:200
of original concentration) and 1.2 ml of sheep blood (Colorado
Senun Company # CS-1112) (blood diluted in a 1:1 buffer providing a
3% concentration of blood per 20 ml medium sample) were added to 20
ml sample of media and mixed. The mixed media preparation was
poured into a petri dished and allowed to solidify.
[0157] Organism Preparation
[0158] Prior to inoculation of petri plates, bacteria or cultures
were examined for morphology under a phase microscope to confirm
motility and physical characteristics, e.g., spiral shaped. The
organisms were washed off the plate into PBS (Phosphate buffered
saline) (80.0 g NaCl, 2.0 g KCl, 2.4 g KH.sub.2PO.sub.4 11.5 g
Na.sub.2HPO.sub.4 or 24.4 g Na.sub.2HPO.sub.4/2H.sub.2O per 1 liter
deionized water, pH 7.2, sterilized by autoclaving) and diluted
into BHI at a concentration of 10.sup.8/ml (O.D. of 0.3 units
diluted 1:30). Ten microliters of solution containing 10.sup.6
organisms were spread onto a petri plate and grown under
microaerophilic conditions for 3 days unless specified. Control
organisms were grown under microaerophilic conditions.
[0159] Results
[0160] Inhibition of growth was determined by the amount of
colonies present within the petri plate. The following factors were
used for growth inhibition: .+-.=0-20 colonies, +=20-50 colonies,
++=50-200 colonies and +++=200+ colonies.
2TABLE I H. pylori ATCC 43504 controls: Organisms + 0.5% DMSO No
growth Organisms + 0.05% DMSO (+++) Organisms without DMSO (+++) A
stock solution containing 10 .mu.g/ml of therapeutic compound and
0.5% DMSO was prepared for each therapeutic compound. Serial
dilutions were prepared from each stock solution. Final drug
Concentration Monensin Nigericin 10 .mu.g/ml No growth No growth 1
.mu.g/ml No growth No growth 100 ng/ml No growth (++) 10 ng/ml (+)
(+++) 1 ng/ml (+) (+++) 0.1 ng/ml (+++) (+++)
[0161]
3TABLE II H. pylori (urease positive) ATCC 43504 controls:
Organisms + 0.5% DMSO (+++) Organisms + 0.05% DMSO (+++) Organisms
without DMSO (+++) A stock solution containing 10 .mu.g/ml of
therapeutic compound and 0.5% DMSO was prepared for each
therapeutic compound. Serial dilutions were prepared from each
stock solution. Final drug Concentration Monensin Nigericin 10
.mu.g/ml No growth No growth 1 .mu.g/ml +/- No growth 100 ng/ml
(+++) 10 ng/ml (+++) 1 ng/ml (+++) (+++) 0.1 nglml (+++) (+++)
[0162]
4TABLE III H. pylori (urease negative) (Wild strain of ATCC 43504)
controls: Organisms + 0.5% DMSO (+++) Organisms + 0.05% DMSO (+++)
Organisms without DMSO (+++) A stock solution containing 10
.mu.g/ml of therapeutic compound and 0.5% DMSO was prepared for
each therapeutic compound. Serial dilutions were prepared from each
stock solution. Final drug Concentration Monensin Nigericin 10
.mu.g/ml No growth No growth 1 .mu.g/ml No growth No growth 100
ng/ml (+++) (+++) 10 ng/ml (+++) (+++) 1 ng/ml (+++) 0.1 ng/ml
(+++) (+++)
[0163]
5TABLE IV H. pylori (urease positive) ATCC 43504 controls:
Organisms + 0.5% DMSO (+++) Organisms + 0.05% DMSO (+++) Organisms
without DMSO (++) A stock solution containing 10 .mu.g/ml of
therapeutic compound and 0.5% DMSO was prepared for each
therapeutic compound. Serial dilutions were prepared from each
stock solution. Final drug Concentration Monensin Nigericin 10
.mu.g/ml No growth No growth 1 .mu.g/ml No growth No growth 100
ng/ml (+++) (+++) 10 ng/ml (+++) (+++) 1 ng/ml (+++) (+++) 0.1
ng/ml (+++) (+++)
[0164]
6TABLE V H. pylori ATCC 43504 Controls: Organisms + 0.5% DMSO (+++)
Organisms + 0.05% DMSO (+++) Organisms without DMSO (+++) A stock
solution containing 10 .mu.g/ml of therapeutic compound and 0.5%
DMSO was prepared for each therapeutic compound. Serial dilutions
were prepared from each stock solution. Assay done in the presence
of 10 mM urea which was added to and mixed with the culture medium
prior to pouring into culture plates. Final drug Concentration
Monensin Nigericin 10 .mu.g/ml No growth No growth.sup.1 1 .mu.g/ml
No growth (.+-.).sup.1 100 ng/ml (+++) (+++).sup.1 10 ng/ml (+++)
(++).sup.1 1 ng/ml (+++) (+++).sup.1 0.1 ng/ml (+++) (+++).sup.1
.sup.1 = assay done in duplicate
[0165]
7TABLE VI H. pylori ATCC 43504 Control: Organisms + 0.05% DMSO
(+++) Organisms without DMSO (+++) A stock solution containing 1
.mu.g/ml of therapeutic compound and 0.05% DMSO was prepared for
each therapeutic compound. Serial dilutions were prepared from each
stock solution. Assay done in the presence of 2.5 mM urea prepared
as above at a lower concentration of urea. Final drug Concentration
Monensin Nigericin 1 .mu.g/ml No growth (.+-.) 10 ng/ml (+++) (+++)
1 ng/ml (+++) (+++)
[0166]
8TABLE VII H. pylori (urease positive) ATCC 43504 controls:
Organisms + 0.5% DMSO (+++) Organisms + 0.05% DMSO (+++) Organisms
without DMSO (+++) A stock solution containing 10 .mu.g/ml of
therapeutic compound and 0.5% DMSO was prepared for each
therapeutic compound. Serial dilutions were prepared from each
stock solution. Final drug Concentration Lasalocid Salinomycin 10
.mu.g/ml (+++) 7. 1.56 .mu.g/ml (+++) 8. 0.78 .mu.g/ml (+++) 9.
0.39 .mu.g/ml (+++) 10. 0.195 .mu.g/ml (+++) 11. 0.098 .mu.g/ml
(+++) MIC = 6.25 .mu.g/ml
[0167]
9TABLE VIII H. pylori (urease positive) ATCC Number 51652 controls:
Organisms + 0.5% ethanol (+++) Organisms without ethanol (+++) A
stock solution containing 100 .mu.g/ml of therapeutic compound and
0.5% DMSO was prepared for each therapeutic compound. Serial
dilutions were prepared from each stock solution. Final drug
concentration Monensin 1. 100 .mu.g/ml No growth 2. 50 .mu.g/ml No
growth 3. 25 .mu.g/ml No growth 4. 12.5 .mu.g/ml No growth 5. 6.25
.mu.g/ml No growth 6. 3.125 .mu.g/ml No growth 7. 1.56 .mu.g/ml No
growth 8. 0.78 .mu.g/ml No growth 9. 0.39 .mu.g/ml No growth 10.
0.195 .mu.g/ml No growth 11. 0.098 .mu.g/ml No growth
[0168]
10TABLE IX H. pylori ATCC 43504 controls: Organisms + 0.5% ethanol
(+++) Organisms without ethanol (+++) A stock solution containing
100 .mu.g/ml of therapeutic compound and 0.5% DMSO was prepared for
each therapeutic compound. Serial dilutions were prepared from each
stock solution. Final drug concentration Fmoc-Ala-monensin 1. 100
.mu.g/ml No growth 2. 50 .mu.g/ml No growth 3. 25 .mu.g/ml No
growth 4. 12.5 .mu.g/ml No growth 5. 6.25 .mu.g/ml (.+-.) 6. 3.125
.mu.g/ml (+++) 7. 1.56 .mu.g/ml (+++) 8. 0.78 .mu.g/ml (+++) 9.
0.39 .mu.g/ml (+++) 10. 0.195 .mu.g/ml (+++) 11. 0.098 .mu.g/ml
(+++) MIC = 6.25 .mu.g/ml
[0169]
11TABLE X H. pylori ATCC 43504 controls: Organisms + 0.5% ethanol
(+++) Organisms without ethanol (+++) A stock solution containing
100 .mu.g/ml of therapeutic compound and 0.5% DMSO was prepared for
each therapeutic compound. Serial dilutions were prepared from each
stock solution. Final drug Monensin phenyl concentration carbamate
1. 100 .mu.g/ml No growth 2. 50 .mu.g/ml No growth 3. 25 .mu.g/ml
No growth 4. 12.5 .mu.g/ml No growth 5. 6.25 .mu.g/ml No growth 6.
3.125 .mu.g/ml (.+-.) few small colonies 7. 1.56 .mu.g/ml (+) small
colonies 8. 0.78 .mu.g/ml (++) 9. 0.39 .mu.g/ml (++) 10. 0.195
.mu.g/ml (+++) 11. 0.098 .mu.g/ml (+++) MIC = 1.56 .mu.g/ml
[0170]
12TABLE XI H. pylori ATCC 43504 controls: Organisms + 0.5% ethanol
(+++) Organisms without ethanol (+++) A stock solution containing
100 .mu.g/ml of therapeutic compound and 0.5% DMSO was prepared for
each therapeutic compound. Serial dilutions were prepared from each
stock solution. Final drug Monensin pyridyl concentration carbamate
1. 100 .mu.g/ml No growth 2. 50 .mu.g/ml No growth 3. 25 .mu.g/ml
No growth 4. 12.5 .mu.g/ml No growth 5. 6.25 .mu.g/ml No growth 6.
3.125 .mu.g/ml (.+-.) 7. 1.56 .mu.g/ml (++) 8. 0.78 .mu.g/ml (+++)
9. 0.39 .mu.g/ml (+++) 10. 0.195 .mu.g/ml (+++) 11. 0.098 .mu.g/ml
(+++) MIC = 3.12 .mu.g/ml
[0171]
13TABLE XII H. pylori ATCC 43504 controls: Organisms + 0.5% ethanol
(+++) Organisms without ethanol (+++) A stock solution containing
100 .mu.g/ml of therapeutic compound and 0.5% DMSO was prepared for
each therapeutic compound. Serial dilutions were prepared from each
stock solution. Final drug concentration Benzyl Monensin 1. 100
.mu.g/ml (.+-.) 2. 50 .mu.g/ml (.+-.) 3. 25 .mu.g/ml (.+-.) 4. 12.5
.mu.g/ml (++) 5. 6.25 .mu.g/ml (+++) 6. 3.125 .mu.g/ml (+++) 7.
1.56 .mu.g/ml (+++) 8. 0.78 .mu.g/ml (+++) 9. 0.39 .mu.g/ml (+++)
10. 0.195 .mu.g/ml (+++) 11. 0.098 .mu.g/ml (+++) MIC = 25
.mu.g/ml
[0172]
14TABLE XIII H. pylori ATCC 43504 controls: Organisms + 0.5%
ethanol (+++) Organisms without ethanol (+++) A stock solution
containing 100 .mu.g/ml of therapeutic compound and 0.5% DMSO was
prepared for each therapeutic compound. Serial dilutions were
prepared from each stock solution. Final drug concentration
Monensin 1. 100 .mu.g/ml No growth 2. 50 .mu.g/ml No growth 3. 25
.mu.g/ml No growth 4. 12.5 .mu.g/ml No growth 5. 6.25 .mu.g/ml No
growth 8. 0.78 .mu.g/ml No growth 9. 0.39 .mu.g/ml (.+-.) 10. 0.195
.mu.g/ml (++) 11. 0.098 .mu.g/ml (+++) MIC = 0.39 .mu.g/ml
[0173]
15TABLE XIV Staph ATCC 14775 controls: Organisms + 0.5% DMSO (+++)
Organisms + 0.05% DMSO (+++) Organisms without DMSO (+++) A stock
solution containing 10 .mu.g/ml of therapeutic compound and 0.5%
DMSO was prepared for each therapeutic compound. Serial dilutions
were prepared from each stock solution. Non-microaerophilic
conditions Final drug Concentration Monensin Nigericin 10 .mu.g/ml
No growth No growth 1 .mu.g/ml (+++***) No growth 100 ng/ml (+++)
(+++***) 10 ng/ml (+++) (+++) 1 ng/ml (+++) (+++) 0.1 ng/ml (+++)
(+++) *** = good growth, but the colonies are smaller than in
controls.
[0174]
16TABLE XV Staph. ATCC 14775 controls: Organisms + 0.5% DMSO (+++)
Organisms + 0.05% DMSO (+++) Organisms without DMSO (+++) A stock
solution containing 10 .mu.g/ml of therapeutic compound and 0.5%
DMSO was prepared for each therapeutic compound. Serial dilutions
were prepared from each stock solution. Non microaerophilic
conditions Final drug concentration Lasalocid Salinomycin 10
.mu.g/ml No growth No growth 1 .mu.g/ml (+++) (+++).sup.1 100 ng/ml
(+++) (+++) 10 ng/ml (+++) (+++) .sup.1small colonies
[0175]
17TABLE XVI Staph. ATCC 14775 controls: Organisms + 0.5% ethanol
(+++) Organisms + 0.25% ethanol (+++) Organisms + 0.125% ethanol
(+++) Organisms without ethanol (+++) A stock solution containing
100 .mu.g/ml of therapeutic compound and 0.5% DMSO was prepared for
each therapeutic compound. Serial dilutions were prepared from each
stock solution. Non-aerophilic conditions. Final drug concentration
Monensin 1. 100 .mu.g/ml No growth 2. 50 .mu.g/ml No growth 3. 25
.mu.g/ml (.+-.).sup.1 4. 12.5 .mu.g/ml (.+-.).sup.1 5. 6.25
.mu.g/ml (+).sup.2 6. 3.125 .mu.g/ml (+).sup.2 7. 1.56 .mu.g/ml
(+).sup.2 8. 0.78 .mu.g/ml (++) 9. 0.39 .mu.g/ml (+++) 10. 0.195
.mu.g/ml (+++) 11. 0.098 .mu.g/ml (+++) (.+-.).sup.1 = very small
colonies, (+).sup.2 = small colonies
[0176]
18TABLE XVII H. pylori (urease positive) ATCC Number 51652 Assay
done in Blood agar base + 3% sheep blood controls: Organisms + 0.5%
DMSO (+++) Organisms + 0.05% DMSO (+++) Organisms without DMSO
(+++) A stock solution containing 10 .mu.g/ml of therapeutic
compound and 0.5% DMSO was prepared for each therapeutic compound.
Serial dilutions were prepared from each stock solution. Lasalocid
Salinomycin 10 .mu.g/ml (+++) No growth 1 .mu.g/ml (+++) (+++) 100
ng/ml (+++) (+++) 10 ng/ml (+++) (+++) 1 ng/ml (+++) (+++) 0.1
ng/ml (+++) (+++) Final drug Concentration Ionomycin A23187 10
.mu.g/ml* (+++) No growth 1 .mu.g/ml** (+++) No growth 100 ng/ml
(+++) (+++) 10 ng/ml (+++) (+++) 1 ng/ml (+++) (+++) 0.1 ng/ml
(+++) (+++)
EQUIVALENTS
[0177] Those skilled in the art will recognize, or be able to
ascertain, using no more than routine experimentation, many
equivalents to specific embodiments of the invention described
specifically herein. Such equivalents are intended to be
encompassed in the scope of the following claims.
* * * * *