U.S. patent application number 12/473955 was filed with the patent office on 2010-05-06 for 9-aminomethyl substituted minocycline compounds.
This patent application is currently assigned to Paratek Pharmaceuticals, Inc.. Invention is credited to Beena Bhatia, Todd Bowser, Roger Frechette, Laura Honeyman, Mohamed Y. Ismail, Mark L. Nelson.
Application Number | 20100113400 12/473955 |
Document ID | / |
Family ID | 46171045 |
Filed Date | 2010-05-06 |
United States Patent
Application |
20100113400 |
Kind Code |
A1 |
Nelson; Mark L. ; et
al. |
May 6, 2010 |
9-Aminomethyl Substituted Minocycline Compounds
Abstract
The present invention pertains, at least in part, to novel
9-substituted minocycline compounds. These minocycline compounds
can be used to treat numerous tetracycline compound-responsive
states, such as bacterial infections and neoplasms, as well as
other known applications for minocycline and minocycline compounds
in general, such as blocking tetracycline efflux and modulation of
gene expression.
Inventors: |
Nelson; Mark L.; (Norfolk,
MA) ; Frechette; Roger; (Reading, MA) ;
Ismail; Mohamed Y.; (Bedford, MA) ; Honeyman;
Laura; (Roslindale, MA) ; Bowser; Todd;
(Charlton, MA) ; Bhatia; Beena; (Arlington,
MA) |
Correspondence
Address: |
MCCARTER & ENGLISH, LLP BOSTON
265 Franklin Street
Boston
MA
02110
US
|
Assignee: |
Paratek Pharmaceuticals,
Inc.
Boston
MA
|
Family ID: |
46171045 |
Appl. No.: |
12/473955 |
Filed: |
May 28, 2009 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
10786881 |
Feb 24, 2004 |
7553828 |
|
|
12473955 |
|
|
|
|
10412656 |
Apr 10, 2003 |
|
|
|
10786881 |
|
|
|
|
10384855 |
Mar 10, 2003 |
|
|
|
10412656 |
|
|
|
|
09895857 |
Jun 29, 2001 |
6846939 |
|
|
10412656 |
|
|
|
|
60395495 |
Jul 12, 2002 |
|
|
|
60362654 |
Mar 8, 2002 |
|
|
|
60275621 |
Mar 13, 2001 |
|
|
|
Current U.S.
Class: |
514/152 ;
549/439; 552/205 |
Current CPC
Class: |
C07C 237/26 20130101;
C07D 317/66 20130101 |
Class at
Publication: |
514/152 ;
552/205; 549/439 |
International
Class: |
A61K 31/65 20060101
A61K031/65; C07C 237/26 20060101 C07C237/26; C07D 317/46 20060101
C07D317/46 |
Claims
1. A compound of formula I: ##STR00045## wherein X is
CHC(R.sup.13Y'Y), CR.sup.6'R.sup.6, S, NR.sup.6, or O; R.sup.2,
R.sup.4', R.sup.4'', R.sup.7' and R.sup.7'' are each hydrogen,
alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl,
alkylsulfonyl, alkylamino, arylalkyl, aryl, heterocyclic,
heteroaromatic or a prodrug moiety; R.sup.3, R.sup.10, R.sup.11 and
R.sup.12 are each hydrogen or a pro-drug moiety; R.sup.4 is
NR.sup.4'R.sup.4'', alkyl, alkenyl, alkynyl, aryl, hydroxyl,
halogen, or hydrogen; R.sup.5 is hydroxyl, hydrogen, thiol,
alkanoyl, aroyl, alkaroyl, aryl, heteroaromatic, alkyl, alkenyl,
alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,
alkylamino, arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy;
R.sup.6 and R.sup.6' are independently hydrogen, methylene, absent,
hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl; R.sup.8 is hydrogen, hydroxyl, halogen, thiol, alkyl,
alkenyl, alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl,
alkylsulfonyl, alkylamino, or an arylalkyl; R.sup.9 is aminoalkyl;
R.sup.13 is hydrogen, hydroxy, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl; Y' and Y are each independently hydrogen, halogen,
hydroxyl, cyano, sulfhydryl, amino, alkyl, alkenyl, alkynyl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl, and pharmaceutically acceptable salts, esters and
prodrugs thereof.
2. The compound of claim 1, wherein R.sup.4 is NR.sup.4'R.sup.4'';
X is CR.sup.6R.sup.6'; R.sup.2, R.sup.2', R.sup.5, R.sup.6,
R.sup.6', R.sup.8, R.sup.9, R.sup.10, R.sup.11 and R.sup.12 are
each hydrogen; and, R.sup.4', R.sup.4'', R.sup.7' and R.sup.7'' are
each lower alkyl.
3. The compound of claim 2, wherein R.sup.4', R.sup.4'', R.sup.7',
and R.sup.7'' are each methyl.
4. The compound of claim 3, wherein said aminoalkyl is
aminomethyl.
5. The compound of claim 3 or 4, wherein said aminoalkyl is
substituted with an alkyl group.
6.-15. (canceled)
16. The compound of claim 1, wherein said aminoalkyl is of the
formula --CH.sub.2NR.sup.9cC(=Z')ZR.sup.9a, wherein Z is
CR.sup.9dR.sup.9e, S, NR.sup.9b or O; Z' is NR.sup.9f, O or S; and
R.sup.9a, R.sup.9b, R.sup.9c, R.sup.9d, R.sup.9e and R.sup.9f are
each independently hydrogen, acyl, alkyl, alkenyl, alkynyl, alkoxy,
alkoxyalkyl, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino,
arylalkyl, aryl, heterocyclic, heteroaromatic or a prodrug
moiety;
17.-25. (canceled)
26. The compound of claim 1, wherein said compound is selected from
the group consisting of: ##STR00046## ##STR00047## ##STR00048## and
pharmaceutically acceptable salts thereof.
27. A compound of formula (II): ##STR00049## wherein: J.sup.5 and
J.sup.6 are each independently hydrogen, alkyl, alkenyl, alkynyl,
aryl, sulfonyl, acyl, alkoxycarbonyl, alkaminocarbonyl,
alkaminothiocarbonyl, substituted thiocarbonyl, substituted
carbonyl, alkoxythiocarbonyl, or linked to form a ring; J.sup.7 and
J.sup.8 are each alkyl, halogen, or hydrogen; X is
CHC(R.sup.13Y'Y), CR.sup.6'R.sup.6, C.dbd.CR.sup.6'R.sup.6, S,
NR.sup.6, or O; R.sup.2, R.sup.2', R.sup.4', and R.sup.4'' are each
independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,
alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic, heteroaromatic or a prodrug moiety; R.sup.4 is
NR.sup.4'R.sup.4'', alkyl, alkenyl, alkynyl, aryl, hydroxyl,
halogen, or hydrogen; R.sup.3, R.sup.10, R.sup.11 and R.sup.12a are
each hydrogen or a pro-drug moiety; R.sup.5 is hydroxyl, hydrogen,
thiol, alkanoyl, aroyl, alkaroyl, aryl, heteroaromatic, alkyl,
alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,
alkylamino, arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy;
R.sup.6 and R.sup.6' are each independently hydrogen, methylene,
absent, hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl; R.sup.7 and R.sup.8 are each independently hydrogen,
dialkylamino, hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl,
aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino,
or an arylalkyl; R.sup.13 is hydrogen, hydroxy, alkyl, alkenyl,
alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,
alkylamino, or an arylalkyl; and Y' and Y are each independently
hydrogen, halogen, hydroxyl, cyano, sulfhydryl, amino, alkyl,
alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,
alkylamino, or an arylalkyl, and pharmaceutically acceptable salts
thereof.
28. The compound of claim 27, wherein R.sup.4 is
NR.sup.4'R.sup.4'', X is CR.sup.6R.sup.6'; R.sup.2, R.sup.2',
R.sup.6, R.sup.6', R.sup.8, R.sup.10, R.sup.11, and R.sup.12 are
each hydrogen; R.sup.4' and R.sup.4'' are lower alkyl; R.sup.7 is
dialkylamino; and R.sup.5 is hydroxy or hydrogen.
29. The compound of claim 28, wherein R.sup.4' and R.sup.4'' are
each methyl and R.sup.5 is hydrogen.
30. The compound of claim 28, wherein J.sup.7 and J.sup.8 are
hydrogen.
31. The compound of claim 28, wherein J.sup.5 is substituted or
unsubstituted alkyl.
32.-34. (canceled)
35. The compound of claim 33, wherein J.sup.6 is hydrogen.
36. A compound of the formula (III): ##STR00050## wherein J.sup.5
is alkyl; and J.sup.6 is hydrogen, or pharmaceutically acceptable
salts, prodrugs and esters thereof.
37. (canceled)
38. A method for treating a tetracycline responsive state in a
mammal, comprising administering to said subject a compound of
claim 1, such that said subject is treated.
39.-47. (canceled)
48. A pharmaceutical composition comprising a therapeutically
effective amount of a compound of claim 1, and a pharmaceutically
acceptable carrier.
Description
RELATED APPLICATIONS
[0001] This application is a continuation application of U.S.
patent application Ser. No. 10/786,881, filed on Feb. 24, 2004,
issuing; which is a continuation of U.S. patent application Ser.
No. 10/412,656, filed on Apr. 10, 2003, abandoned; which is a
continuation-in-part of Ser. No. 10/384,855, filed on Mar. 10,
2003, abandoned, which claims priority to U.S. Provisional Patent
Application Ser. No. 60/395,495, filed on Jul. 12, 2002; and U.S.
Provisional Patent Application Ser. No. 60/362,654, filed Mar. 8,
2002. U.S. patent application Ser. No. 10/412,656 is a
continuation-in-part of U.S. patent application Ser. No.
09/895,857, now U.S. Pat. No. 6,846,939, issued Jan. 25, 2005,
which claims priority to U.S. Patent Application Ser. No.
60/275,621, filed on Mar. 13, 2001. The entire contents of each of
the aforementioned applications are incorporated herein by
reference in their entirety.
BACKGROUND OF THE INVENTION
[0002] The development of the tetracycline antibiotics was the
direct result of a systematic screening of soil specimens collected
from many parts of the world for evidence of microorganisms capable
of producing bacteriocidal and/or bacteriostatic compositions. The
first of these novel compounds was introduced in 1948 under the
name chlortetracycline. Two years later, oxytetracycline became
available. The elucidation of the chemical structure of these
compounds confirmed their similarity and furnished the analytical
basis for the production of a third member of this group in 1952,
tetracycline. A new family of minocycline compounds, without the
ring-attached methyl group present in earlier tetracyclines, was
prepared in 1957 and became publicly available in 1967; and
minocycline was in use by 1972. Recently, research efforts have
focused on developing new tetracycline antibiotic compositions
effective under varying therapeutic conditions and routes of
administration. New tetracycline analogues have also been
investigated which may prove to be equal to or more effective than
the originally introduced minocycline compounds. Examples include
U.S. Pat. Nos. 2,980,584; 2,990,331; 3,062,717; 3,165,531;
3,454,697; 3,557,280; 3,674,859; 3,957,980; 4,018,889; 4,024,272;
and 4,126,680. These patents are representative of the range of
pharmaceutically active tetracycline and tetracycline analogue
compositions.
[0003] Historically, soon after their initial development and
introduction, the tetracyclines were found to be highly effective
pharmacologically against rickettsiae; a number of gram-positive
and gram-negative bacteria; and the agents responsible for
lymphogranuloma venereum, inclusion conjunctivitis, and
psittacosis. Hence, tetracyclines became known as "broad spectrum"
antibiotics. With the subsequent establishment of their in vitro
antimicrobial activity, effectiveness in experimental infections,
and pharmacological properties, the tetracyclines as a class
rapidly became widely used for therapeutic purposes. However, this
widespread use of tetracyclines for both major and minor illnesses
and diseases let directly to
the emergence of resistance to these antibiotics even among highly
susceptible bacterial species both commensal and pathogenic (e.g.,
pneumococci and Salmonella). The rise of tetracycline-resistant
organisms has resulted in a general decline in use of tetracyclines
and tetracycline analogue compositions as antibiotics of
choice.
SUMMARY OF THE INVENTION
[0004] The invention pertains, at least in part, to compounds of
formula I:
##STR00001##
wherein:
[0005] X is CHC(R.sup.13Y'Y), CR.sup.6'R.sup.6, S, NR.sup.6, or
O;
[0006] R.sup.2, R.sup.4', R.sup.4'', R.sup.7' and R.sup.7'' are
each hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,
alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic, heteroaromatic or a prodrug moiety;
[0007] R.sup.4 is NR.sup.4'R.sup.4'', alkyl, alkenyl, alkynyl,
aryl, hydroxyl, halogen, or hydrogen;
[0008] R.sup.2', R.sup.3, R.sup.10, R.sup.11 and R.sup.12 are each
hydrogen or a pro-drug moiety;
[0009] R.sup.5 is hydroxyl, hydrogen, thiol, alkanoyl, aroyl,
alkanoyl, aryl, heteroaromatic, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl,
alkyl carbonyloxy, or aryl carbonyloxy;
[0010] R.sup.6 and R.sup.6' are independently hydrogen, methylene,
absent, hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl;
[0011] R.sup.8 is hydrogen, hydroxyl, halogen, thiol, alkyl,
alkenyl, alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl,
alkylsulfonyl, alkylamino, or an arylalkyl;
[0012] R.sup.9 is aminoalkyl;
[0013] R.sup.13 is hydrogen, hydroxy, alkyl, alkenyl, alkynyl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl;
[0014] Y' and Y are each independently hydrogen, halogen, hydroxyl,
cyano, sulfhydryl, amino, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl, and pharmaceutically acceptable salts, esters and
prodrugs thereof.
[0015] The invention also pertains, at least in part, to compounds
of formula (II):
##STR00002##
wherein:
[0016] J.sup.5 and J.sup.6 are each independently hydrogen, alkyl,
alkenyl, alkynyl, aryl, sulfonyl, acyl, alkoxycarbonyl,
alkaminocarbonyl, alkaminothiocarbonyl, substituted thiocarbonyl,
substituted carbonyl, alkoxythiocarbonyl, or linked to form a
ring;
[0017] J.sup.7 and J.sup.8 are each alkyl, halogen, or
hydrogen;
[0018] X is CHC(R.sup.13Y'Y), CR.sup.6'R.sup.6,
C.dbd.CR.sup.6'R.sup.6, S, NR.sup.6, or O;
[0019] R.sup.2, R.sup.2', R.sup.4', and R.sup.4'' are each
independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,
alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic, heteroaromatic or a prodrug moiety;
[0020] R.sup.4 is NR.sup.4'R.sup.4'', alkyl, alkenyl, alkynyl,
aryl, hydroxyl, halogen, or hydrogen;
[0021] R.sup.2', R.sup.3, R.sup.10, R.sup.11 and R.sup.12 are each
hydrogen or a pro-drug moiety;
[0022] R.sup.5 is hydroxyl, hydrogen, thiol, alkanoyl, aroyl,
alkaroyl, aryl, heteroaromatic, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl,
alkyl carbonyloxy, or aryl carbonyloxy;
[0023] R.sup.6 and R.sup.6' are each independently hydrogen,
methylene, absent, hydroxyl, halogen, thiol, alkyl, alkenyl,
alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,
alkylamino, or an arylalkyl;
[0024] R.sup.7 and R.sup.8 and are each independently hydrogen,
hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl;
[0025] R.sup.13 is hydrogen, hydroxy, alkyl, alkenyl, alkynyl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl; and
[0026] Y' and Y are each independently hydrogen, halogen, hydroxyl,
cyano, sulfhydryl, amino, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl, and pharmaceutically acceptable salts thereof.
[0027] A compound of the formula (III):
##STR00003##
[0028] wherein [0029] J.sup.5 is alkyl; and [0030] J.sup.6 is
hydrogen, or pharmaceutically acceptable salts, prodrugs and esters
thereof.
[0031] The invention also pertains to pharmaceutical compositions
comprising the compounds of the invention (e.g., compounds of
formula (I), (II), (III), or otherwise described herein) and a
pharmaceutically acceptable carrier. The invention also pertains to
the use of a compound of the invention for the manufacture of a
medicament, e.g., a medicament for the treatment of a tetracycline
responsive state.
[0032] The invention also pertains to methods of using the
compounds of the invention to treat subjects suffering from
tetracycline compound responsive states.
DETAILED DESCRIPTION OF THE INVENTION
[0033] The present invention pertains, at least in part, to novel
9-substituted minocycline compounds. These minocycline compounds
can be used to treat numerous tetracycline compound-responsive
states, such as bacterial infections and neoplasms, as well as
other known applications for minocycline and minocycline compounds
in general, such as blocking tetracycline efflux and modulation of
gene expression.
[0034] The invention pertains, at least in part, to minocycline
compounds of Formula I:
##STR00004##
wherein:
[0035] X is CHC(R.sup.13Y'Y), CR.sup.6'R.sup.6, S, NR.sup.6, or
O;
[0036] R.sup.2, R.sup.4', R.sup.4'', R.sup.7' and R.sup.7'' are
each hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,
alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic, heteroaromatic or a prodrug moiety;
[0037] R.sup.4 is NR.sup.4'R.sup.4'', alkyl, alkenyl, alkynyl,
aryl, hydroxyl, halogen, or hydrogen;
[0038] R.sup.2', R.sup.3, R.sup.10, R.sup.11 and R.sup.12 are each
hydrogen or a pro-drug moiety;
[0039] R.sup.5 is hydroxyl, hydrogen, thiol, alkanoyl, aroyl,
alkaroyl, aryl, heteroaromatic, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl,
alkyl carbonyloxy, or aryl carbonyloxy;
[0040] R.sup.6 and R.sup.6' are independently hydrogen, methylene,
absent, hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl;
[0041] R.sup.8 is hydrogen, hydroxyl, halogen, thiol, alkyl,
alkenyl, alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl,
alkylsulfonyl, alkylamino, or arylalkyl;
[0042] R.sup.9 is aminoalkyl;
[0043] R.sup.13 is hydrogen, hydroxy, alkyl, alkenyl, alkynyl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl;
[0044] Y' and Y are each independently hydrogen, halogen, hydroxyl,
cyano, sulfhydryl, amino, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, allylamino, or an
arylalkyl, and pharmaceutically acceptable salts, esters and
prodrugs thereof.
[0045] The term minocycline compounds refers to compounds of
formulae (I), (II), and (III) above. In an embodiment, the term
minocycline compounds include compounds wherein X is
CR.sup.6R.sup.6'; R.sup.2, R.sup.2', R.sup.5, R.sup.6, R.sup.6',
R.sup.9, R.sup.10, R.sup.11, and R.sup.12 are each hydrogen;
R.sup.4 is NR.sup.4R.sup.4''; and R.sup.4', R.sup.4'', R.sup.7',
and R.sup.7'' are each lower alkyl, e.g., methyl. Other compounds
of the invention include compounds wherein R.sup.4 is hydrogen.
[0046] The invention pertains, at least in part, to compounds of
Formula (I) wherein R.sup.9 is aminoalkyl (e.g., aminomethyl, e.g.,
--CH.sub.2NR'R''). Aminoalkyl R.sup.9 groups may be further
substituted. Examples of substituents include alkyl, alkenyl, aryl,
alkynyl, carbonyl, and acyl groups. Examples of aryl groups include
such as, for example substituted or unsubstituted phenyl (e.g.,
methylenedioxyphenyl or para-perfluoromethoxyphenyl), or
heteroaromatic groups which allows the compound of the invention to
perform its intended function. Alkyl groups include methyl, ethyl,
i-propyl, n-propyl, i-butyl, n-butyl, t-butyl, pentyl, hexyl,
heptyl, octyl, nonyl, decyl, etc. Cyclic alkyl groups include
groups with one or more rings, such as, for example, cyclopropane,
cyclobutane, cyclopentane, cyclohexane, cycloheptane, etc. In an
embodiment, the alkyl R.sup.9 group is 2-cyclopentylethyl.
[0047] Examples of substituents of alkyl groups include, for
example, halogens (e.g., fluorine, chlorine, bromine, iodine,
etc.), hydroxyl, alkoxy (e.g., methoxy, ethoxy, propoxy, butoxy,
pentoxy, perfluoromethoxy, perchloromethoxy, etc.),
alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, allcylcarbonyl,
alkylaminoacarbonyl, arylalkyl aminocarbonyl, alkenylaminocarbonyl,
carboxy, alkylcarbonyl, arylcarbonyl, arylalkylcarbonyl,
alkenylcarbonyl, alkoxycarbonyl, silyl, aminocarbonyl,
alkylthiocarbonyl, phosphate, phosphonato, phosphinato, cyano,
amino, acylamino, amido, imino, sulfhydryl, alkylthio, arylthio,
thiocarboxylate, sulfate, alkylsulfinyl, alkenyl, sulfonato,
sulfamoyl, sulfonamido, nitro, alkenyl, cyano, azido, heterocyclyl,
alkylaryl, aryl and heteroaryl.
[0048] In a further embodiment, the minocycline compound is
selected from the group consisting of:
##STR00005##
and pharmaceutically acceptable salts, esters and prodrugs
thereof.
[0049] In another embodiment, the minocycline compound of the
invention is a compound wherein R.sup.9 is
--CH.sub.2NR.sup.9cC(=Z')ZR.sup.9a, wherein Z is CR.sup.9dR.sup.9e,
S, NR.sup.9b or O; Z' is NR.sup.9f, O or S; and R.sup.9a, R.sup.9b,
R.sup.9c, R.sup.9d, and R.sup.9f are each independently hydrogen,
acyl, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfonyl,
alkylsulfonyl, alkylamino, arylalkyl, aryl, heterocyclic,
heteroaromatic or a prodrug moiety.
[0050] In certain embodiments, R.sup.9 is
--CH.sub.2NR.sup.9cC(=Z')ZR.sup.9a. Examples of R.sup.9c include
hydrogen. Z' may be, for example, S, NH, or O. Examples of Z
include NR.sup.9b (e.g., when R.sup.9b is hydrogen, alkyl, etc.), O
or S.
[0051] Examples of R.sup.9a groups include aryl groups such as
substituted and unsubstituted phenyl. Examples of possible
substituents of aryl R.sup.9a groups include, but are not limited
to, alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, hexyl,
perfluormethyl, perchloroethyl, etc.), alkenyl, halogen (e.g.,
fluorine, chlorine, bromine, iodine, etc.), hydroxyl, alkoxy (e.g.,
methoxy, ethoxy, propoxy, perfluoromethoxy, perchloromethoxy,
etc.), alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
alkylaminoacarbonyl, arylalkyl aminocarbonyl, alkenylaminocarbonyl,
alkylcarbonyl, arylcarbonyl, arylalkylcarbonyl, alkenylcarbonyl,
alkoxycarbonyl, silyl, aminocarbonyl, alkylthiocarbonyl, phosphate,
phosphonato, phosphinato, cyano, amino, acylamino, amido, imino,
sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfate,
alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro, acetyl,
alkyl, cyano, azido, heterocyclyl, alkylaryl, aryl and heteroaryl
groups.
[0052] In certain embodiments, at least one of the substituents of
the substituted phenyl is nitro, alkoxy (e.g., methoxy,
methylenedioxy, perfluoromethoxy) alkyl (e.g., methyl, ethyl,
propyl, butyl, or pentyl), acetyl, halogen (e.g., fluorine,
chlorine, bromine, or iodine), or amino (e.g., dialkylamino). In
certain embodiments, the alkoxy group is perhalogenated, e.g.,
perfluoromethoxy.
[0053] Examples of aryl R.sup.9a groups include, but are not
limited to, unsubstituted phenyl, para-nitrophenyl, para-methoxy
phenyl, para-perfluoromethoxy phenyl, para-acetyl phenyl,
3,5-methylenedioxyphenyl, 3,5-diperfluoromethyl phenyl, para-bromo
phenyl, para-chloro phenyl, and para-fluoro phenyl.
[0054] Other examples of aryl R.sup.9a groups include substituted
and unsubstituted heterocycles (e.g., furanyl, imidazolyl,
benzothiophenyl, benzofuranyl, quinolinyl, isoquinolinyl,
benzodioxazolyl, benzoxazolyl, benzothiazolyl, benzoimidazolyl,
methylenedioxyphenyl, indolyl, thienyl, pyrimidyl, pyrazinyl,
purinyl, pyrazolyl, pyrrolidinyl, oxazolyl, isooxazolyl,
naphthridinyl, thiazolyl, isothiazolyl, or deazapurinyl) and
substituted and unsubstituted biaryl groups, such as naphthyl and
fluorene.
[0055] R.sup.9a also may be substituted or unsubstituted alkyl,
e.g., methyl, ethyl, propyl, butyl, pentyl, etc. Examples of
substituents include but are not limited to halogens (e.g.,
fluorine, bromine, chlorine, iodine, etc.), hydroxyl, alkoxy (e.g.,
methoxy, ethoxy, propoxy, butoxy, etc.), alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, alkylaminoacarbonyl, arylalkyl
aminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl, arylcarbonyl,
arylalkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl, silyl,
aminocarbonyl, alkylthiocarbonyl, phosphate, phosphonato,
phosphinato, cyano, amino, acylamino, amidino, imino, sulfhydryl,
alkylthio, arylthio, thiocarboxylate, sulfate, alkylsulfinyl,
sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano,
azido, alkenyl, heterocyclyl, alkylaryl, aryl and heteroaryl.
[0056] R.sup.9a also can be substituted or unsubstituted alkenyl.
Examples of substituents for alkenyl R.sup.9a groups include those
listed above for alkyl R.sup.9a groups. Examples of alkenyl
R.sup.9a groups include pent-1-enyl.
[0057] In an embodiment, Z' is NH, Z is NH, and R.sup.9a is
alkyl.
[0058] In a further embodiment, the minocycline compound of the
invention does not include compounds wherein R.sup.9 is
dimethylmaleimido, when R.sup.9 is dimethylmaleimido or when the
compound is described in Martell et al. (J. Med. Chem., (1967,
10(3), 359-3). In another embodiment, the minocycline compounds of
the invention do not include compounds wherein R.sup.9 is
4-morpholinylmethyl, when R.sup.7 is hydrogen or when the compound
is described in Strel'nokov (Antibiotiki, (1965), 10(7), 650-6),
Polyak (Ref. Zh., Biol. Khim. Abstr. No. 6F782), Paikin, M. D.
(Ref. Zh., Farmakol, Toksikol. 1965, Abstr. No. 5.54.323), In
another embodiment, the compounds of the invention do not include
compounds wherein R.sup.9 is 1-pyrrolidinylmethyl, when R.sup.7 is
hydrogen, or compounds otherwise described in Genazzini, E. et al.
(Atti. Soc. Ital. Sci. Vet. (1964), 18, 175-8, Hajdu, P.
Arzneimittel-Forsch. (1962), 12, 206-7, Federal Register, (1962),
27, 3851, Baldini et al. Boll. Soc. Ital. Biol. Sper. (1960), 36,
577-83), Good, W., Biochim. et Biophys., Acta, (1962) 56, 359-61;
Ritzerfeld, W., Arzneimittel-Forsch, (1962), 12, 30-2; Maniar, A.
et al., Ann. Inst. Pasteur, (1961), 101, 887-97); Garrod, L.,
Recenti Progr. Med. (1962), 32, 3-24; Branceni, Compt. Rend. Soc.
Biol. (1961), 155, 1469-72, ES 302929, In another embodiment, the
compounds of the invention do not include compounds wherein R.sup.9
is --CH.sub.2NHCH.sub.2C(.dbd.O)NH.sub.2,
--CH.sub.2NHCH(CH.sub.3)C(O)H(CH.sub.2).sub.2OH,
--CH.sub.2NHCH(CH.sub.3)C(.dbd.O)NH.sub.2,
--CH.sub.2NHCH.sub.2C(.dbd.O)NHCH.sub.3,
--CH.sub.2NHCH.sub.2C(.dbd.O)NH(CH.sub.2).sub.2OH, or
--CH.sub.2NHCH(C(.dbd.O)NH(CH.sub.2).sub.2)(CH.sub.2).sub.4NH.sub.2,
or --CH.sub.2NHCH(C(.dbd.O)(CH.sub.2).sub.4NH.sub.2, when R.sup.7
is H, or otherwise described in GB921252 or GB 955766.
[0059] Examples of minocycline compounds of the invention include
those listed in Table 1, as well as the ones listed below:
##STR00006## ##STR00007##
[0060] The invention also pertains, at least in part, to
minocycline compounds of formula (II):
##STR00008##
[0061] wherein:
[0062] J.sup.5 and J.sup.6 are each independently hydrogen, alkyl,
alkenyl, alkynyl, aryl, sulfonyl, acyl, alkoxycarbonyl,
alkaminocarbonyl, alkaminothiocarbonyl, substituted thiocarbonyl,
substituted carbonyl, alkoxythiocarbonyl, or linked to form a
ring;
[0063] J.sup.7 and J.sup.8 are each alkyl, halogen, or
hydrogen;
[0064] X is CHC(R.sup.13Y'Y), CR.sup.6'R.sup.6,
C.dbd.CR.sup.6'R.sup.6, S, NR.sup.6, or O;
[0065] R.sup.2, R.sup.2', R.sup.4', and R.sup.4'' are each
independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,
alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic, heteroaromatic or a prodrug moiety;
[0066] R.sup.4 is NR.sup.4'R.sup.4'', alkyl, alkenyl, alkynyl,
aryl, hydroxyl, halogen, or hydrogen;
[0067] R.sup.2', R.sup.3, R.sup.10, R.sup.11 and R.sup.12 are each
hydrogen or a pro-drug moiety;
[0068] R.sup.5 is hydroxyl, hydrogen, thiol, alkanoyl, aroyl,
alkaroyl, aryl, heteroaromatic, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl,
alkyl carbonyloxy, or aryl carbonyloxy;
[0069] R.sup.6 and R.sup.6' are each independently hydrogen,
methylene, absent, hydroxyl, halogen, thiol, alkyl, alkenyl,
alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,
alkylamino, or an arylalkyl;
[0070] R.sup.8 and R.sup.7 are each independently hydrogen,
hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl;
[0071] R.sup.13 is hydrogen, hydroxy, alkyl, alkenyl, alkynyl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl; and
[0072] Y' and Y are each independently hydrogen, halogen, hydroxyl,
cyano, sulfhydryl, amino, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an
arylalkyl, and pharmaceutically acceptable salts thereof.
[0073] In one embodiment, R.sup.4' and R.sup.4'' are each methyl
and R.sup.5 is hydrogen. In another further embodiment, J.sup.7 and
J.sup.8 are hydrogen. In another embodiment, J.sup.5 is substituted
or unsubstituted alkyl, e.g., branched or straight chain alkyl. In
another embodiment, J.sup.5 is methyl, ethyl, propyl, pentyl,
hexyl, octyl, etc. In a further embodiment, J.sup.5 is n-pentyl. In
another further embodiment, J.sup.6 is hydrogen.
[0074] A compound of the formula (III):
##STR00009##
[0075] wherein [0076] J.sup.5 is alkyl; and [0077] J.sup.6 is
hydrogen, or pharmaceutically acceptable salts, prodrugs and esters
thereof.
[0078] In a further embodiment, J.sup.5 is pentyl, e.g., n-pentyl,
and J.sup.6 is hydrogen.
[0079] The compounds of this invention can be synthesized using the
methods described in Schemes 1-7, in combination with methods known
in the art. Scheme 1 depicts the reaction of sancycline with an
aminoalkylating reagent under appropriate conditions such that an
aminoalkyl minocycline compound is formed.
##STR00010##
[0080] Examples of aminoalkylating reagents, include, but are not
limited to, compounds of the formula (IV):
##STR00011##
[0081] wherein [0082] R.sup.a' and R.sup.a'' are each independently
hydrogen or halogen; [0083] R' is hydrogen, alkyl, alkenyl,
alkynyl, aryl, heteroaryl, or halogen; and [0084] R'' is hydrogen
or optionally linked to R' to form a 4-8 membered ring. The ring
may be optionally substituted, e.g., with halogens and may comprise
carbons and/or heteroatoms such as oxygen, nitrogen, and sulfur. R'
may be further substituted with any substituent which does not
prevent the reagent from reacting with the tetracycline compound of
the invention, under the appropriate conditions. In another further
embodiment, R' is alkyl, e.g., unsubstituted or substituted (e.g.,
with halogens, e.g., chlorine, fluorine, bromine, iodine, etc.). In
another embodiment, R' is aryl, e.g., phenyl, e.g., unsubstituted
or substituted (e.g., with halogens (e.g., chlorine, bromine,
fluorine, etc.), hydroxy, alkoxy, esters, amino, etc.). In another
embodiment, R.sup.a' and R.sup.a'' are each hydrogen. Other
examples of aminoalkylating reagents include
N-hydroxymethylphthalimide.
[0085] Examples of amino-alkylating reagents include, but are not
limited to:
##STR00012##
[0086] The term "appropriate conditions" include those conditions
under which the aminoalkylating reagent and the tetracycline
compound interact such that an aminoalkyl tetracycline compound is
formed. The appropriate conditions may comprise treating the
tetracycline compound with an acid prior to, or concurrently with
the addition of the aminoalkylating reagent to the reaction
mixture. Examples of acids which maybe used alone or in combination
include acids known in the art, as well as, sulfuric acid,
hydrofluoric acid (HF), methanesulfonic acid, trifluoromethane
sulfonic acid, hydrochloric acid, hydrochloric acid in aqueous
ethanol, acetic acid, methanesulfonic acid, and trifluoroacetic
acid (TFA). In a further embodiment, appropriate conditions may
also comprise treating the resulting tetracycline compound with a
reaction quenching agent (e.g., water).
[0087] Scheme 2 shows two aminoalkylations of a minocycline
compound with aminoalkylating reagents which comprise a 5 membered
ring. Similar reactions can be also be carried out using reagents,
with, for example, 6- or 7-membered rings.
##STR00013##
[0088] As shown in Scheme 3 below, the synthesis of
7-monosubstituted aminomethyl tetracyclines may be synthesized
using protecting groups (i.e. the 9-t-butyl protecting group) to be
cleaved using art recognized techniques, such as acid. Examples of
acids which can be used include, but are not limited to, HF,
trifluoroacetic acid (TFA), H.sub.2SO.sub.4 and mixtures thereof.
In this way, regioselective aminomethylation at position 7 is
achieved.
##STR00014##
[0089] In a further embodiment, the appropriate conditions may
further comprise treating the reaction mixture (which may comprise
an intermediate aminoalkyl minocycline compound) with a
derivatizing agent under secondary appropriate conditions such that
the desired aminoalkyl minocycline compound is formed. The
reactions in Scheme 4 are shown for the 9 position, but the
reactions are also applicable to other positions of the minocycline
compound. Additional derivatizing agents and secondary appropriate
conditions may be found, for example, in the chemical literature.
See, for example, R. C. LaRock, Comprehensive Organic
Transformations, (New York: VCH Publishers, Inc., 1989) and
references cited therein. Any reagent that can react with a primary
amine to form a new compound is possible. Examples of some of the
diverse structures are shown in Scheme 4 below.
##STR00015##
[0090] For example, in Scheme 5, an acid chloride derivatizing
agent is added to the reaction mixture to form the desired amide
aminoalkyl minocycline compound (J. Am. Chem. Soc. 71, 2215 (1949);
J. Am. Chem. Soc. 108, 1039 (1986); Org. Syn. Coll. Vol. 4, 339
(1963); Org. Syn. Coll. Vol. 5, 387 (1973)).
##STR00016##
[0091] Scheme 6 depicts the reaction of an intermediate aminoalkyl
minocycline compound with an appropriate sulfonyl chloride
derivatizing agent, such that the desired sulfonamide aminoalkyl
compound is formed (Org. Syn. Coll. Vol. 5, 736, 758 (1973)).
##STR00017##
[0092] Scheme 7 depicts the reaction of a derivatizing agent with
an aminoalkyl tetracycline intermediate to form the resulting
carbamate aminoalkyl minocycline compound.
##STR00018##
[0093] The term "alkyl" includes saturated aliphatic groups,
including straight-chain alkyl groups (e.g., methyl, ethyl, propyl,
butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc.),
branched-chain alkyl groups (isopropyl, tert-butyl, isobutyl,
etc.), cycloalkyl (alicyclic) groups (cyclopropyl, cyclopentyl,
cyclohexyl, cycloheptyl, cyclooctyl), alkyl substituted cycloalkyl
groups, and cycloalkyl substituted alkyl groups. The term alkyl
further includes alkyl groups, which can further include oxygen,
nitrogen, sulfur or phosphorous atoms replacing one or more carbons
of the hydrocarbon backbone. In certain embodiments, a straight
chain or branched chain alkyl has 6 or fewer carbon atoms in its
backbone (e.g., C.sub.1-C.sub.6 for straight chain, C.sub.3-C.sub.6
for branched chain), and more preferably 4 or fewer. Likewise,
preferred cycloalkyls have from 3-8 carbon atoms in their ring
structure, and more preferably have 5 or 6 carbons in the ring
structure. The term C.sub.1-C.sub.6 includes alkyl groups
containing 1 to 6 carbon atoms.
[0094] Moreover, the term alkyl includes 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, alkenyl, alkynyl, halogen, hydroxyl,
alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, 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, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.
Cycloalkyls can be further substituted, e.g., with the substituents
described above. An "alkylaryl" or an "arylalkyl" moiety is an
alkyl substituted with an aryl (e.g., phenylmethyl (benzyl)). The
term "alkyl" also includes the side chains of natural and unnatural
amino acids.
[0095] The term "aryl" includes groups, including 5- and 6-membered
single-ring aromatic groups that may include from zero to four
heteroatoms, for example, benzene, phenyl, pyrrole, furan,
thiophene, thiazole, isothiaozole, imidazole, triazole, tetrazole,
pyrazole, oxazole, isooxazole, pyridine, pyrazine, pyridazine, and
pyrimidine, and the like. Furthermore, the term "aryl" includes
multicyclic aryl groups, e.g., tricyclic, bicyclic, e.g.,
naphthalene, benzoxazole, benzodioxazole, benzothiazole,
benzoimidazole, benzothiophene, methylenedioxyphenyl, quinoline,
isoquinoline, napthridine, indole, benzofuran, purine, benzofuran,
deazapurine, or indolizine. Those aryl groups having heteroatoms in
the ring structure may also be referred to as "aryl heterocycles",
"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,
alkylaminoacarbonyl, arylalkyl aminocarbonyl, alkenylaminocarbonyl,
alkylcarbonyl, arylcarbonyl, arylalkylcarbonyl, alkenylcarbonyl,
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, alkylsulfinyl, 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).
[0096] The term "alkenyl" includes unsaturated aliphatic groups
analogous in length and possible substitution to the alkyls
described above, but that contain at least one double bond.
[0097] For example, the term "alkenyl" includes straight-chain
alkenyl groups (e.g., ethylenyl, propenyl, butenyl, pentenyl,
hexenyl, heptenyl, octenyl, nonenyl, decenyl, etc.), branched-chain
alkenyl groups, cycloalkenyl (alicyclic) groups (cyclopropenyl,
cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl), alkyl or
alkenyl substituted cycloalkenyl groups, and cycloalkyl or
cycloalkenyl substituted alkenyl groups. The term alkenyl further
includes alkenyl groups which include oxygen, nitrogen, sulfur or
phosphorous atoms replacing one or more carbons of the hydrocarbon
backbone. In certain embodiments, a straight chain or branched
chain alkenyl group has 6 or fewer carbon atoms in its backbone
(e.g., C.sub.2-C.sub.6 for straight chain, C.sub.3-C.sub.6 for
branched chain). Likewise, cycloalkenyl groups may have from 3-8
carbon atoms in their ring structure, and more preferably have 5 or
6 carbons in the ring structure. The term C.sub.2-C.sub.6 includes
alkenyl groups containing 2 to 6 carbon atoms.
[0098] Moreover, the term alkenyl includes both "unsubstituted
alkenyls" and "substituted alkenyls", the latter of which refers to
alkenyl moieties having substituents replacing a hydrogen on one or
more carbons of the hydrocarbon backbone. Such substituents can
include, for example, alkyl groups, alkynyl groups, halogens,
hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, 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, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic
moiety.
[0099] The term "alkynyl" includes unsaturated aliphatic groups
analogous in length and possible substitution to the alkyls
described above, but which contain at least one triple bond.
[0100] For example, the term "alkynyl" includes straight-chain
alkynyl groups (e.g., ethynyl, propynyl, butynyl, pentynyl,
hexynyl, heptynyl, octynyl, nonynyl, decynyl, etc.), branched-chain
alkynyl groups, and cycloalkyl or cycloalkenyl substituted alkynyl
groups. The term alkynyl further includes alkynyl groups which
include oxygen, nitrogen, sulfur or phosphorous atoms replacing one
or more carbons of the hydrocarbon backbone. In certain
embodiments, a straight chain or branched chain alkynyl group has 6
or fewer carbon atoms in its backbone (e.g., C.sub.2-C.sub.6 for
straight chain, C.sub.3-C.sub.6 for branched chain). The term
C.sub.2-C.sub.6 includes alkynyl groups containing 2 to 6 carbon
atoms.
[0101] Moreover, the term alkynyl includes both "unsubstituted
alkynyls" and "substituted alkynyls", the latter of which refers to
alkynyl moieties having substituents replacing a hydrogen on one or
more carbons of the hydrocarbon backbone. Such substituents can
include, for example, alkyl groups, alkynyl groups, halogens,
hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, 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, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic
moiety.
[0102] 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 five carbon atoms in its backbone structure.
"Lower alkenyl" and "lower alkynyl" have chain lengths of, for
example, 2-5 carbon atoms.
[0103] The term "acyl" includes compounds and moieties which
contain the acyl radical (CH.sub.3CO--) or a carbonyl group. The
term "substituted acyl" includes acyl groups where one or more of
the hydrogen atoms are replaced by for example, alkyl groups,
alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,
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, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an
aromatic or heteroaromatic moiety.
[0104] The term "acylamino" includes moieties wherein an acyl
moiety is bonded to an amino group. For example, the term includes
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido
groups.
[0105] The term "aroyl" includes compounds and moieties with an
aryl or heteroaromatic moiety bound to a carbonyl group. Examples
of aroyl groups include phenylcarboxy, naphthyl carboxy, etc.
[0106] The terms "alkoxyalkyl", "alkylaminoalkyl" and
"thioalkoxyalkyl" include alkyl groups, as described above, which
further include oxygen, nitrogen or sulfur atoms replacing one or
more carbons of the hydrocarbon backbone, e.g., oxygen, nitrogen or
sulfur atoms.
[0107] The term "alkoxy" includes substituted and unsubstituted
alkyl, alkenyl, and alkynyl groups covalently linked to an oxygen
atom. Examples of alkoxy groups include methoxy, ethoxy,
isopropyloxy, propoxy, butoxy, and pentoxy groups. Examples of
substituted alkoxy groups include halogenated alkoxy groups. The
alkoxy groups can be substituted with groups such as alkenyl,
alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,
alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, 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, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moieties.
Examples of halogen substituted alkoxy groups include, but are not
limited to, fluoromethoxy, difluoromethoxy, trifluoromethoxy,
chloromethoxy, dichloromethoxy, trichloromethoxy, etc.
[0108] The term "amine" or "amino" includes compounds where a
nitrogen atom is covalently bonded to at least one carbon or
heteroatom. The term "alkyl amino" includes groups and compounds
wherein the nitrogen is bound to at least one additional alkyl
group. The term "dialkyl amino" includes groups wherein the
nitrogen atom is bound to at least two additional alkyl groups. The
term "arylamino" and "diarylamino" include groups wherein the
nitrogen is bound to at least one or two aryl groups, respectively.
The term "alkylarylamino," "alkylaminoaryl" or "arylaminoalkyl"
refers to an amino group which is bound to at least one alkyl group
and at least one aryl group. The term "alkaminoalkyl" refers to an
alkyl, alkenyl, or alkynyl group bound to a nitrogen atom which is
also bound to an alkyl group.
[0109] The term "amide" or "aminocarbonyl" includes compounds or
moieties which contain a nitrogen atom which is bound to the carbon
of a carbonyl or a thiocarbonyl group. The term includes
"alkaminocarbonyl" or "alkylaminocarbonyl" groups which include
alkyl, alkenyl, aryl or alkynyl groups bound to an amino group
bound to a carbonyl group. It includes arylaminocarbonyl groups
which include aryl or heteroaryl moieties bound to an amino group
which is bound to the carbon of a carbonyl or thiocarbonyl group.
The terms "alkylaminocarbonyl," "alkenylaminocarbonyl,"
"alkynylaminocarbonyl," "arylaminocarbonyl," "alkylcarbonylamino,"
"alkenylcarbonylamino," "alkynylcarbonylamino," and
"arylcarbonylamino" are included in term "amide." Amides also
include urea groups (aminocarbonylamino) and carbamates
(oxycarbonylamino).
[0110] The term "carbonyl" or "carboxy" includes compounds and
moieties which contain a carbon connected with a double bond to an
oxygen atom. Examples of moieties which contain a carbonyl include
aldehydes, ketones, carboxylic acids, amides, esters, anhydrides,
etc.
[0111] The term "thiocarbonyl" or "thiocarboxy" includes compounds
and moieties which contain a carbon connected with a double bond to
a sulfur atom.
[0112] The term "ether" includes compounds or moieties which
contain an oxygen bonded to two different carbon atoms or
heteroatoms. For example, the term includes "alkoxyalkyl" which
refers to an alkyl, alkenyl, or alkynyl group covalently bonded to
an oxygen atom which is covalently bonded to another alkyl
group.
[0113] The term "ester" includes compounds and moieties which
contain a carbon or a heteroatom bound to an oxygen atom which is
bonded to the carbon of a carbonyl group. The term "ester" includes
alkoxycarboxy groups such as methoxycarbonyl, ethoxycarbonyl,
propoxycarbonyl, butoxycarbonyl, pentoxycarbonyl, etc. The alkyl,
alkenyl, or alkynyl groups are as defined above.
[0114] The term "thioether" includes compounds and moieties which
contain a sulfur atom bonded to two different carbon or hetero
atoms. Examples of thioethers include, but are not limited to
alkthioalkyls, alkthioalkenyls, and alkthioalkynyls. The term
"alkthioalkyls" include compounds with an alkyl, alkenyl, or
alkynyl group bonded to a sulfur atom which is bonded to an alkyl
group. Similarly, the term "alkthioalkenyls" and alkthioalkynyls"
refer to compounds or moieties wherein an alkyl, alkenyl, or
alkynyl group is bonded to a sulfur atom which is covalently bonded
to an alkynyl group.
The term "hydroxy" or "hydroxyl" includes groups with an --OH or
--O.sup.-.
[0115] The term "halogen" includes fluorine, bromine, chlorine,
iodine, etc. The term "perhalogenated" generally refers to a moiety
wherein all hydrogens are replaced by halogen atoms.
[0116] The terms "polycyclyl" or "polycyclic radical" refer to 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, alkylaminoacarbonyl, arylalkylaminocarbonyl,
alkenylaminocarbonyl, alkylcarbonyl, arylcarbonyl, arylalkyl
carbonyl, alkenylcarbonyl, 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,
alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, alkyl, alkylaryl, or
an aromatic or heteroaromatic moiety.
[0117] The term "heteroatom" includes atoms of any element other
than carbon or hydrogen. Preferred heteroatoms are nitrogen,
oxygen, sulfur and phosphorus.
[0118] The term "prodrug moiety" includes moieties which can be
metabolized in vivo to an active group and moieties which may
advantageously remain attached in vivo. Preferably, the prodrugs
moieties are metabolized in vivo by enzymes, e.g., esterases or by
other mechanisms to hydroxyl groups or other advantageous groups.
Examples of prodrugs and their uses are well known in the art (See,
e.g., Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci.
66:1-19). The prodrugs can be prepared in situ during the final
isolation and purification of the compounds, or by separately
reacting the purified compound with a suitable agent. Hydroxyl
groups can be converted into esters via treatment with a carboxylic
acid. Examples of prodrug moieties include substituted and
unsubstituted, branch or unbranched lower alkyl ester moieties,
(e.g., propionoic acid esters), lower alkenyl esters, di-lower
alkyl-amino lower-alkyl esters (e.g., dimethylaminoethyl ester),
acylamino lower alkyl esters (e.g., acetyloxymethyl ester), acyloxy
lower alkyl esters (e.g., pivaloyloxymethyl ester), aryl esters
(phenyl ester), aryl-lower alkyl esters (e.g., benzyl ester),
substituted (e.g., with methyl, halo, or methoxy substituents) aryl
and aryl-lower alkyl esters, amides, lower-alkyl amides, di-lower
alkyl amides, and hydroxy amides. Preferred prodrug moieties are
propionoic acid esters and acyl esters.
[0119] 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.
Furthermore, the structures and other compounds and moieties
discussed in this application also include all tautomers
thereof.
[0120] The invention also pertains to methods for treating a
tetracycline responsive states in subjects, by administering to a
subject an effective amount of a compound of the invention (e.g., a
compound of Formula (I), (II), (III), or otherwise described
herein), such that the tetracycline responsive state is
treated.
[0121] The language "tetracycline compound responsive state" or
"tetracycline responsive state" includes states which can be
treated, prevented, or otherwise ameliorated by the administration
of a compound of the invention, e.g., a compound of Formula (I),
(II), (III) or otherwise described herein. Tetracycline compound
responsive states include bacterial, viral, and fungal infections
(including those which are resistant to other tetracycline
compounds), cancer (e.g., prostate, breast, colon, lung melanoma
and lymph cancers and other disorders characterized by unwanted
cellular proliferation, including, but not limited to, those
described in U.S. Pat. No. 6,100,248), arthritis, osteoporosis,
diabetes, cystic fibrosis, neurological disorders and other states
for which tetracycline compounds have been found to be active (see,
for example, U.S. Pat. Nos. 5,789,395; 5,834,450; 6,277,061 and
5,532,227, each of which is expressly incorporated herein by
reference). Compounds of the invention can be used to prevent or
control important mammalian and veterinary diseases such as
diarrhea, urinary tract infections, infections of skin and skin
structure, ear, nose and throat infections, wound infection,
mastitis and the like. In addition, methods for treating neoplasms
using tetracycline compounds of the invention are also included
(van der Bozert et al., Cancer Res., 48:6686-6690 (1988)). In a
further embodiment, the tetracycline responsive state is not a
bacterial infection. Other tetracycline compound responsive states
include, for example, those described in U.S. Ser. No.
10/196,010.
[0122] Tetracycline compound responsive states also include
inflammatory process associated states (IPAS). The term
"inflammatory process associated state" includes states in which
inflammation or inflammatory factors (e.g., matrix
metalloproteinases (MMPs), nitric oxide (NO), TNF, interleukins,
plasma proteins, cellular defense systems, cytokines, lipid
metabolites, proteases, toxic radicals, adhesion molecules, etc.)
are involved or are present in an area in aberrant amounts, e.g.,
in amounts which may be advantageous to alter, e.g., to benefit the
subject. The inflammatory process is the response of living tissue
to damage. The cause of inflammation may be due to physical damage,
chemical substances, micro-organisms, tissue necrosis, cancer or
other agents. Acute inflammation is short-lasting, lasting only a
few days. If it is longer lasting however, then it may be referred
to as chronic inflammation.
[0123] IPAS's include inflammatory disorders. Inflammatory
disorders are generally characterized by heat, redness, swelling,
pain and loss of function. Examples of causes of inflammatory
disorders include, but are not limited to, microbial infections
(e.g., bacterial and fungal infections), physical agents (e.g.,
burns, radiation, and trauma), chemical agents (e.g., toxins and
caustic substances), tissue necrosis and various types of
immunologic reactions.
[0124] Examples of inflammatory disorders include, but are not
limited to, osteoarthritis, rheumatoid arthritis, acute and chronic
infections (bacterial and fungal, including diphtheria and
pertussis); acute and chronic bronchitis, sinusitis, and upper
respiratory infections, including the common cold; acute and
chronic gastroenteritis and colitis; acute and chronic cystitis and
urethritis; acute and chronic dermatitis; acute and chronic
conjunctivitis; acute and chronic serositis (pericarditis,
peritonitis, synovitis, pleuritis and tendinitis); uremic
pericarditis; acute and chronic cholecystis; acute and chronic
vaginitis; acute and chronic uveitis; drug reactions; insect bites;
burns (thermal, chemical, and electrical); and sunburn.
[0125] Tetracycline compound responsive states also include NO
associated states. The term "NO associated state" includes states
which involve or are associated with nitric oxide (NO) or inducible
nitric oxide synthase (iNOS). NO associated state includes states
which are characterized by aberrant amounts of NO and/or iNOS.
Preferably, the NO associated state can be treated by administering
tetracycline compounds of the invention, e.g., compounds of formula
I, II, III, or otherwise described herein. The disorders, diseases
and states described in U.S. Pat. Nos. 6,231,894; 6,015,804;
5,919,774; and 5,789,395 are also included as NO associated states.
The entire contents of each of these patents are hereby
incorporated herein by reference.
[0126] Other examples of NO associated states include, but are not
limited to, malaria, senescence, diabetes, vascular stroke,
neurodegenerative disorders (Alzheimer's disease, Huntington's
disease), cardiac disease (reperfusion-associated injury following
infarction), juvenile diabetes, inflammatory disorders,
osteoarthritis, rheumatoid arthritis, acute and chronic infections
(bacterial, viral, and fungal); cystic fibrosis, acute and chronic
bronchitis, sinusitis, and respiratory infections, including the
common cold; acute and chronic gastroenteritis and colitis; acute
and chronic cystitis and urethritis; acute and chronic dermatitis;
acute and chronic conjunctivitis; acute and chronic serositis
(pericarditis, peritonitis, synovitis, pleuritis and tendinitis);
uremic pericarditis; acute and chronic cholecystis; acute and
chronic vaginitis; acute and chronic uveitis; drug reactions;
insect bites; burns (thermal, chemical, and electrical); and
sunburn.
[0127] The term "inflammatory process associated state" also
includes, in one embodiment, matrix metalloproteinase associated
states (MMPAS). MMPAS include states characterized by aberrant
amounts of MMPs or MMP activity. These are also include as
tetracycline compound responsive states which may be treated using
compounds of the invention, e.g., in formula (I), (II), (III) or
otherwise described herein.
[0128] Examples of matrix metalloproteinase associated states
("MMPAS's") include, but are not limited to, arteriosclerosis,
corneal ulceration, emphysema, osteoarthritis, multiple sclerosis
(Liedtke et al., Ann. Neurol. 1998, 44:35-46; Chandler et al., J.
Neuroimmunol. 1997, 72:155-71), osteosarcoma, osteomyelitis,
bronchiectasis, chronic pulmonary obstructive disease, skin and eye
diseases, periodontitis, osteoporosis, rheumatoid arthritis,
ulcerative colitis, inflammatory disorders, tumor growth and
invasion (Stetler-Stevenson et al., Annu. Rev. Cell Biol. 1993,
9:541-73; Tryggvason et al., Biochim. Biophys. Acta 1987,
907:191-217; Li et al., Mol. Carcinog. 1998, 22:84-89), metastasis,
acute lung injury, stroke, ischemia, diabetes, aortic or vascular
aneurysms, skin tissue wounds, dry eye, bone and cartilage
degradation (Greenwald et al., Bone 1998, 22:33-38; Ryan et al.,
Curr. Op. Rheumatol. 1996, 8; 238-247). Other MMPAS include those
described in U.S. Pat. Nos. 5,459,135; 5,321,017; 5,308,839;
5,258,371; 4,935,412; 4,704,383, 4,666,897, and RE 34,656,
incorporated herein by reference in their entirety.
[0129] In another embodiment, the tetracycline compound responsive
state is cancer. Examples of cancers which the tetracycline
compounds of the invention may be useful to treat include all solid
tumors, i.e., carcinomas e.g., adenocarcinomas, and sarcomas.
Adenocarcinomas are carcinomas derived from glandular tissue or in
which the tumor cells form recognizable glandular structures.
Sarcomas broadly include tumors whose cells are embedded in a
fibrillar or homogeneous substance like embryonic connective
tissue. Examples of carcinomas which may be treated using the
methods of the invention include, but are not limited to,
carcinomas of the prostate, breast, ovary, testis, lung, colon, and
breast. The methods of the invention are not limited to the
treatment of these tumor types, but extend to any solid tumor
derived from any organ system. Examples of treatable cancers
include, but are not limited to, colon cancer, bladder cancer,
breast cancer, melanoma, ovarian carcinoma, prostatic carcinoma,
lung cancer, and a variety of other cancers as well. The methods of
the invention also cause the inhibition of cancer growth in
adenocarcinomas, such as, for example, those of the prostate,
breast, kidney, ovary, testes, and colon.
[0130] In an embodiment, the tetracycline responsive state of the
invention is cancer. The invention pertains to a method for
treating a subject suffering or at risk of suffering from cancer,
by administering an effective amount of a substituted tetracycline
compound, such that inhibition cancer cell growth occurs, i.e.,
cellular proliferation, invasiveness, metastasis, or tumor
incidence is decreased, slowed, or stopped. The inhibition may
result from inhibition of an inflammatory process, down-regulation
of an inflammatory process, some other mechanism, or a combination
of mechanisms. Alternatively, the tetracycline compounds may be
useful for preventing cancer recurrence, for example, to treat
residual cancer following surgical resection or radiation therapy.
The tetracycline compounds useful according to the invention are
especially advantageous as they are substantially non-toxic
compared to other cancer treatments. In a further embodiment, the
compounds of the invention are administered in combination with
standard cancer therapy, such as, but not limited to,
chemotherapy.
[0131] The language "in combination with" another therapeutic agent
or treatment includes co-administration of the tetracycline
compound and with the other therapeutic agent or treatment,
administration of the tetracycline compound first, followed by the
other therapeutic agent or treatment and administration of the
other therapeutic agent or treatment first, followed by the
tetracycline compound. The other therapeutic agent may be any agent
which is known in the art to treat, prevent, or reduce the symptoms
of a tetracycline responsive state. Furthermore, the other
therapeutic agent may be any agent of benefit to the patient when
administered in combination with the administration of an
tetracycline compound. In one embodiment, the cancers treated by
methods of the invention include those described in U.S. Pat. No.
6,100,248; 5,843,925; 5,837,696; or 5,668,122, incorporated herein
by reference in their entirety.
[0132] In another embodiment, the tetracycline compound responsive
state is diabetes, e.g., juvenile diabetes, diabetes mellitus,
diabetes type I, diabetes type II, diabetic ulcers, or other
diabetic complications. In a further embodiment, protein
glycosylation is not affected by the administration of the
tetracycline compounds of the invention. In another embodiment, the
tetracycline compound of the invention is administered in
combination with standard diabetic therapies, such as, but not
limited to insulin therapy. In a further embodiment, the IPAS
includes disorders described in U.S. Pat. Nos. 5,929,055; and
5,532,227, incorporated herein by reference in their entirety.
[0133] In another embodiment, the tetracycline compound responsive
state is a bone mass disorder. Bone mass disorders include
disorders where a subjects bones are disorders and states where the
formation, repair or remodeling of bone is advantageous. For
examples bone mass disorders include osteoporosis (e.g., a decrease
in bone strength and density), bone fractures, bone formation
associated with surgical procedures (e.g., facial reconstruction),
osteogenesis imperfecta (brittle bone disease), hypophosphatasia,
Paget's disease, fibrous dysplasia, osteopetrosis, myeloma bone
disease, and the depletion of calcium in bone, such as that which
is related to primary hyperparathyroidism. Bone mass disorders
include all states in which the formation, repair or remodeling of
bone is advantageous to the subject as well as all other disorders
associated with the bones or skeletal system of a subject which can
be treated with the tetracycline compounds of the invention. In a
further embodiment, the bone mass disorders include those described
in U.S. Pat. Nos. 5,459,135; 5,231,017; 5,998,390; 5,770,588; RE
34,656; U.S. Pat. Nos. 5,308,839; 4,925,833; 3,304,227; and
4,666,897, each of which is hereby incorporated herein by reference
in its entirety.
[0134] In another embodiment, the tetracycline compound responsive
state is acute lung injury. Acute lung injuries include adult
respiratory distress syndrome (ARDS), post-pump syndrome (PPS), and
trauma. Trauma includes any injury to living tissue caused by an
extrinsic agent or event. Examples of trauma include, but are not
limited to, crush injuries, contact with a hard surface, or cutting
or other damage to the lungs.
[0135] The invention also pertains to a method for treating acute
lung injury by administering a tetracycline compound of the
invention.
[0136] The tetracycline responsive states of the invention also
include chronic lung disorders. The invention pertains to methods
for treating chronic lung disorders by administering a tetracycline
compound, such as those described herein. The method includes
administering to a subject an effective amount of a substituted
tetracycline compound such that the chronic lung disorder is
treated. Examples of chronic lung disorders include, but are not
limited, to asthma, cystic fibrosis, and emphysema. In a further
embodiment, the tetracycline compounds of the invention used to
treat acute and/or chronic lung disorders such as those described
in U.S. Pat. Nos. 5,977,091; 6,043,231; 5,523,297; and 5,773,430,
each of which is hereby incorporated herein by reference in its
entirety.
[0137] In yet another embodiment, the tetracycline compound
responsive state is ischemia, stroke, or ischemic stroke. The
invention also pertains to a method for treating ischemia, stroke,
or ischemic stroke by administering an effective amount of a
substituted tetracycline compound of the invention. In a further
embodiment, the compounds of the invention are used to treat such
disorders as described in U.S. Pat. No. 6,231,894; 5,773,430;
5,919,775 or 5,789,395, incorporated herein by reference.
[0138] In another embodiment, the tetracycline compound responsive
state is a skin wound. The invention also pertains, at least in
part, to a method for improving the healing response of the
epithelialized tissue (e.g., skin, mucosae) to acute traumatic
injury (e.g., cut, burn, scrape, etc.). The method may include
using a tetracycline compound of the invention (which may or may
not have antibacterial activity) to improve the capacity of the
epithelialized tissue to heal acute wounds. The method may increase
the rate of collagen accumulation of the healing tissue. The method
may also decrease the proteolytic activity in the epthithelialized
tissue by decreasing the collagenolytic and/or gelatinolytic
activity of MMPs. In a further embodiment, the tetracycline
compound of the invention is administered to the surface of the
skin (e.g., topically). In a further embodiment, the tetracycline
compound of the invention is used to treat a skin wound, and other
such disorders as described in, for example, U.S. Pat. Nos.
5,827,840; 4,704,383; 4,935,412; 5,258,371; 5,308,8391 5,459,135;
5,532,227; and 6,015,804; each of which is incorporated herein by
reference in its entirety.
[0139] Examples of tetracycline responsive states also include
neurological disorders which include both neuropsychiatric and
neurodegenerative disorders, but are not limited to, such as
Alzheimer's disease, dementias related to Alzheimer's disease (such
as Pick's disease), Parkinson's and other Lewy diffuse body
diseases, senile dementia, Huntington's disease, Gilles de la
Tourette's syndrome, multiple sclerosis, amyotrophic lateral
sclerosis (ALS), progressive supranuclear palsy, epilepsy, and
Creutzfeldt-Jakob disease; autonomic function disorders such as
hypertension and sleep disorders, and neuropsychiatric disorders,
such as depression, schizophrenia, schizoaffective disorder,
Korsakoff's psychosis, mania, anxiety disorders, or phobic
disorders; learning or memory disorders, e.g., amnesia or
age-related memory loss, attention deficit disorder, dysthymic
disorder, major depressive disorder, mania, obsessive-compulsive
disorder, psychoactive substance use disorders, anxiety, phobias,
panic disorder, as well as bipolar affective disorder, e.g., severe
bipolar affective (mood) disorder (BP-1), bipolar affective
neurological disorders, e.g., migraine and obesity. Further
neurological disorders include, for example, those listed in the
American Psychiatric Association's Diagnostic and Statistical
manual of Mental Disorders (DSM), the most current version of which
is incorporated herein by reference in its entirety.
[0140] In yet another embodiment, the tetracycline compound
responsive state is an aortic or vascular aneurysm in vascular
tissue of a subject (e.g., a subject having or at risk of having an
aortic or vascular aneurysm, etc.). The tetracycline compound may
by effective to reduce the size of the vascular aneurysm or it may
be administered to the subject prior to the onset of the vascular
aneurysm such that the aneurysm is prevented. In one embodiment,
the vascular tissue is an artery, e.g., the aorta, e.g., the
abdominal aorta. In a further embodiment, the tetracycline
compounds of the invention are used to treat disorders described in
U.S. Pat. Nos. 6,043,225 and 5,834,449, incorporated herein by
reference in their entirety.
[0141] Bacterial infections may be caused by a wide variety of gram
positive and gram negative bacteria. The compounds of the invention
are useful as antibiotics against organisms which are resistant to
other tetracycline compounds. The antibiotic activity of the
tetracycline compounds of the invention may be determined using the
method discussed in Example 2, or by using the in vitro standard
broth dilution method described in Waitz, J. A., National
Commission for Clinical Laboratory Standards, Document M7-A2, vol.
10, no. 8, pp. 13-20, 2.sup.nd edition, Villanova, Pa. (1990).
[0142] The tetracycline compounds of the invention may also be used
to treat infections traditionally treated with tetracycline
compounds such as, for example, rickettsiae; a number of
gram-positive and gram-negative bacteria; and the agents
responsible for lymphogranuloma venereum, inclusion conjunctivitis,
psittacosis. The tetracycline compounds may be used to treat
infections of, e.g., K. pneumoniae, Salmonella, E. hirae, A.
baumanii, B. catarrhalis, H. influenzae, P. aeruginosa, E. faecium,
E. coli, S. aureus or E. faecalis. In one embodiment, the
tetracycline compound is used to treat a bacterial infection that
is resistant to other tetracycline antibiotic compounds. The
tetracycline compound of the invention may be administered with a
pharmaceutically acceptable carrier.
[0143] The language "effective amount" of the compound is that
amount necessary or sufficient to treat or prevent a tetracycline
compound responsive state. The effective amount can vary depending
on such factors as the size and weight of the subject, the type of
illness, or the particular compound. For example, the choice of the
compound can affect what constitutes an "effective amount". One of
ordinary Skill in the art would be able to study the aforementioned
factors and make the determination regarding the effective amount
of the tetracycline compound without undue experimentation.
[0144] The invention also pertains to methods of treatment against
microorganism infections and associated diseases. The methods
include administration of an effective amount of one or more
minocycline compounds to a subject. The subject can be either a
plant or, advantageously, an animal, e.g., a mammal, e.g., a
human.
[0145] In the therapeutic methods of the invention, one or more
minocycline compounds of the invention may be administered alone to
a subject, or more typically a compound of the invention will be
administered as part of a pharmaceutical composition in mixture
with conventional excipient, i.e., pharmaceutically acceptable
organic or inorganic carrier substances suitable for parenteral,
oral or other desired administration and which do not deleteriously
react with the active compounds and are not deleterious to the
recipient thereof.
[0146] The invention also pertains to pharmaceutical compositions
comprising a therapeutically effective amount of a minocycline
compound and, optionally, a pharmaceutically acceptable
carrier.
[0147] The language "pharmaceutically acceptable carrier" includes
substances capable of being coadministered with the minocycline
compound(s), and which allow both to perform their intended
function, e.g., treat or prevent a tetracycline responsive state.
Suitable pharmaceutically acceptable carriers include but are not
limited to water, salt solutions, alcohol, vegetable oils,
polyethylene glycols, gelatin, lactose, amylose, magnesium
stearate, talc, silicic acid, viscous paraffin, perfume oil, fatty
acid monoglycerides and diglycerides, petroethral fatty acid
esters, hydroxymethyl-cellulose, polyvinylpyrrolidone, etc. The
pharmaceutical preparations can be sterilized and if desired mixed
with auxiliary agents, e.g., lubricants, preservatives,
stabilizers, wetting agents, emulsifiers, salts for influencing
osmotic pressure, buffers, colorings, flavorings and/or aromatic
substances and the like which do not deleteriously react with the
active compounds of the invention.
[0148] The minocycline compounds of the invention that are basic in
nature are capable of forming a wide variety of salts with various
inorganic and organic acids. The acids that may be used to prepare
pharmaceutically acceptable acid addition salts of the minocycline
compounds of the invention that are basic in nature are those that
form non-toxic acid addition salts, i.e., salts containing
pharmaceutically acceptable anions, such as the hydrochloride,
hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate,
acid phosphate, isonicotinate, acetate, lactate, salicylate,
citrate, acid citrate, tartrate, pantothenate, bitartrate,
ascorbate, succinate, maleate, gentisinate, fumarate, gluconate,
glucaronate, saccharate, formate, benzoate, glutamate,
methanesulfonate, ethanesulfonate, benzenesulfonate,
p-toluenesulfonate and palmoate [i.e.,
1,1'-methylene-bis-(2-hydroxy-3-naphthoate)]salts. Although such
salts must be pharmaceutically acceptable for administration to a
subject, e.g., a mammal, it is often desirable in practice to
initially isolate a minocycline compound of the invention from the
reaction mixture as a pharmaceutically unacceptable salt and then
simply convert the latter back to the free base compound by
treatment with an alkaline reagent and subsequently convert the
latter free base to a pharmaceutically acceptable acid addition
salt. The acid addition salts of the base compounds of this
invention are readily prepared by treating the base compound with a
substantially equivalent amount of the chosen mineral or organic
acid in an aqueous solvent medium or in a suitable organic solvent,
such as methanol or ethanol. Upon careful evaporation of the
solvent, the desired solid salt is readily obtained. The
preparation of other minocycline compounds of the invention not
specifically described in the foregoing experimental section can be
accomplished using combinations of the reactions described above
that will be apparent to those skilled in the art.
[0149] The preparation of other minocycline compounds of the
invention not specifically described in the foregoing experimental
section can be accomplished using combinations of the reactions
described above that will be apparent to those skilled in the
art.
[0150] The minocycline compounds of the invention that are acidic
in nature are capable of forming a wide variety of base salts. The
chemical bases that may be used as reagents to prepare
pharmaceutically acceptable base salts of those minocycline
compounds of the invention that are acidic in nature are those that
form non-toxic base salts with such compounds. Such non-toxic base
salts include, but are not limited to those derived from such
pharmaceutically acceptable cations such as alkali metal cations
(e.g., potassium and sodium) and alkaline earth metal cations
(e.g., calcium and magnesium), ammonium or water-soluble amine
addition salts such as N-methylglucamine-(meglumine), and the lower
alkanolammonium and other base salts of pharmaceutically acceptable
organic amines. The pharmaceutically acceptable base addition salts
of minocycline compounds of the invention that are acidic in nature
may be formed with pharmaceutically acceptable cations by
conventional methods. Thus, these salts may be readily prepared by
treating the minocycline compound of the invention with an aqueous
solution of the desired pharmaceutically acceptable cation and
evaporating the resulting solution to dryness, preferably under
reduced pressure. Alternatively, a lower alkyl alcohol solution of
the minocycline compound of the invention may be mixed with an
alkoxide of the desired metal and the solution subsequently
evaporated to dryness.
[0151] The preparation of other minocycline compounds of the
invention not specifically described in the foregoing experimental
section can be accomplished using combinations of the reactions
described above that will be apparent to those skilled in the
art.
[0152] The compounds of the invention and pharmaceutically
acceptable salts thereof can be administered via either the oral,
parenteral or topical routes. In general, these compounds are most
desirably administered in effective dosages, depending upon the
weight and condition of the subject being treated and the
particular route of administration chosen. Variations may occur
depending upon the species of the subject being treated and its
individual response to said medicament, as well as on the type of
pharmaceutical formulation chosen and the time period and interval
at which such administration is carried out.
[0153] The pharmaceutical compositions of the invention may be
administered alone or in combination with other known compositions
for treating tetracycline responsive states in a subject, e.g., a
mammal. Preferred mammals include pets (e.g., cats, dogs, ferrets,
etc.), farm animals (cows, sheep, pigs, horses, goats, etc.), lab
animals (rats, mice, monkeys, etc.), and primates (chimpanzees,
humans, gorillas). The language "in combination with" a known
composition is intended to include simultaneous administration of
the composition of the invention and the known composition,
administration of the composition of the invention first, followed
by the known composition and administration of the known
composition first, followed by the composition of the invention.
Any of the therapeutically composition known in the art for
treating tetracycline responsive states can be used in the methods
of the invention.
[0154] The compounds of the invention may be administered alone or
in combination with pharmaceutically acceptable carriers or
diluents by any of the routes previously mentioned, and the
administration may be carried out in single or multiple doses. For
example, the novel therapeutic agents of this invention can be
administered advantageously in a wide variety of different dosage
forms, i.e., they may be combined with various pharmaceutically
acceptable inert carriers in the form of tablets, capsules,
lozenges, troches, hard candies, powders, sprays, creams, salves,
suppositories, jellies, gels, pastes, lotions, ointments, aqueous
suspensions, injectable solutions, elixirs, syrups, and the like.
Such carriers include solid diluents or fillers, sterile aqueous
media and various non-toxic organic solvents, etc. Moreover, oral
pharmaceutical compositions can be suitably sweetened and/or
flavored. In general, the therapeutically-effective compounds of
this invention are present in such dosage forms at concentration
levels ranging from about 5.0% to about 70% by weight.
[0155] For oral administration, tablets containing various
excipients such as microcrystalline cellulose, sodium citrate,
calcium carbonate, dicalcium phosphate and glycine may be employed
along with various disintegrants such as starch (and preferably
corn, potato or tapioca starch), alginic acid and certain complex
silicates, together with granulation binders like
polyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally,
lubricating agents such as magnesium stearate, sodium lauryl
sulfate and talc are often very useful for tabletting purposes.
Solid compositions of a similar type may also be employed as
fillers in gelatin capsules; preferred materials in this connection
also include lactose or milk sugar as well as high molecular weight
polyethylene glycols. When aqueous suspensions and/or elixirs are
desired for oral administration, the active ingredient may be
combined with various sweetening or flavoring agents, coloring
matter or dyes, and, if so desired, emulsifying and/or suspending
agents as well, together with such diluents as water, ethanol,
propylene glycol, glycerin and various like combinations
thereof.
[0156] For parenteral administration (including intraperitoneal,
subcutaneous, intravenous, intradermal or intramuscular injection),
solutions of a therapeutic compound of the present invention in
either sesame or peanut oil or in aqueous propylene glycol may be
employed. The aqueous solutions should be suitably buffered
(preferably pH greater than 8) if necessary and the liquid diluent
first rendered isotonic. These aqueous solutions are suitable for
intravenous injection purposes. The oily solutions are suitable for
intraarticular, intramuscular and subcutaneous injection purposes.
The preparation of all these solutions under sterile conditions is
readily accomplished by standard pharmaceutical techniques well
known to those skilled in the art. For parenteral application,
examples of suitable preparations include solutions, preferably
oily or aqueous solutions as well as suspensions, emulsions, or
implants, including suppositories. Therapeutic compounds may be
formulated in sterile form in multiple or single dose formats such
as being dispersed in a fluid carrier such as sterile physiological
saline or 5% saline dextrose solutions commonly used with
injectables.
[0157] Additionally, it is also possible to administer the
compounds of the present invention topically when treating
inflammatory conditions of the skin. Examples of methods of topical
administration include transdermal, buccal or sublingual
application. For topical applications, therapeutic compounds can be
suitably admixed in a pharmacologically inert topical carrier such
as a gel, an ointment, a lotion or a cream. Such topical carriers
include water, glycerol, alcohol, propylene glycol, fatty alcohols,
triglycerides, fatty acid esters, or mineral oils. Other possible
topical carriers are liquid petrolatum, isopropylpalmitate,
polyethylene glycol, ethanol 95%, polyoxyethylene monolauriate 5%
in water, sodium lauryl sulfate 5% in water, and the like. In
addition, materials such as anti-oxidants, humectants, viscosity
stabilizers and the like also may be added if desired.
[0158] For enteral application, particularly suitable are tablets,
dragees or capsules having talc and/or carbohydrate carrier binder
or the like, the carrier preferably being lactose and/or corn
starch and/or potato starch. A syrup, elixir or the like can be
used wherein a sweetened vehicle is employed. Sustained release
compositions can be formulated including those wherein the active
component is protected with differentially degradable coatings,
e.g., by microencapsulation, multiple coatings, etc.
[0159] In addition to treatment of human subjects, the therapeutic
methods of the invention also will have significant veterinary
applications, e.g. for treatment of livestock such as cattle,
sheep, goats, cows, swine and the like; poultry such as chickens,
ducks, geese, turkeys and the like; horses; and pets such as dogs
and cats. Also, the compounds of the invention may be used to treat
non-animal subjects, such as plants.
[0160] It will be appreciated that the actual preferred amounts of
active compounds used in a given therapy will vary according to the
specific compound being utilized, the particular compositions
formulated, the mode of application, the particular site of
administration, etc. Optimal administration rates for a given
protocol of administration can be readily ascertained by those
skilled in the art using conventional dosage determination tests
conducted with regard to the foregoing guidelines.
[0161] In general, compounds of the invention for treatment can be
administered to a subject in dosages used in prior tetracycline
therapies. See, for example, the Physicians' Desk Reference. For
example, a suitable effective dose of one or more compounds of the
invention will be in the range of from 0.01 to 100 milligrams per
kilogram of body weight of recipient per day, preferably in the
range of from 0.1 to 50 milligrams per kilogram body weight of
recipient per day, more preferably in the range of 1 to 20
milligrams per kilogram body weight of recipient per day. The
desired dose is suitably administered once daily, or several
sub-doses, e.g. 2 to 5 sub-doses, are administered at appropriate
intervals through the day, or other appropriate schedule.
[0162] It will also be understood that normal, conventionally known
precautions will be taken regarding the administration of
minocyclines generally to ensure their efficacy under normal use
circumstances. Especially when employed for therapeutic treatment
of humans and animals in vivo, the practitioner should take all
sensible precautions to avoid conventionally known contradictions
and toxic effects. Thus, the conventionally recognized adverse
reactions of gastrointestinal distress and inflammations, the renal
toxicity, hypersensitivity reactions, changes in blood, and
impairment of absorption through aluminum, calcium, and magnesium
ions should be duly considered in the conventional manner.
[0163] Furthermore, the invention also pertains to the use of a
compound of formula I, II, III, or otherwise described herein for
the preparation of a medicament. The medicament may include a
pharmaceutically acceptable carrier and the compound is an
effective amount, e.g., an effective amount to treat a tetracycline
responsive state.
Exemplification of the Invention
[0164] Compounds of the invention may be made as described below,
with modifications to the procedure below within the skill of those
of ordinary skill in the art.
Example 1
Synthesis of 9-Aminomethyl Minocycline and Derivatives Thereof
[0165] Trifluoroacetic acid (1 L) was charged into a 2 L flask
under argon and minocycline. HCl (200 g, 1 eq) and
N-hydroxymethylphthalimide (100 g) were added to the flask while
stirring. Once the entire solid dissolved, H.sub.2SO.sub.4 (200 mL)
was added to the reaction. The reaction was heated to 40-50.degree.
C. for 5-6 hours. N-hydroxymethylamine (100 g) was added
portionwise. When HPLC analysis confirmed that all the starting
material was converted to
2,9-bis-aminomethylphthalimidominocycline, the mixture was
precipitated out of 4 L of acetone. An exotherm of 15-20.degree. C.
was observed. After 1 hour of stirring, the solid was filtered,
washed with acetone (200 ml), and dried with the aid of a latex
rubber dam. The solid was reslurried in a methanol (1 L)/t-BME (2
L) mixture and the pH was adjusted to 3 using triethylamine. The
solid was filtered and washed with 50 mL of methanol. The yield was
97% of 2,9-bis-aminomethylphthalimideminocycline.
[0166] 2,9-bis-aminomethylphthalimideminocycline (100 g) was
suspended in 2M solution of methylamine in methanol (10 eq). The
reaction was stirred at room temperature for 2-3 hours, at which
point HPLC analysis confirmed total conversion of the starting
material to 2,9-bis aminomethylminocycline. The reaction mixture
was poured into t-BME (5 volumes), and stirred for thirty minutes.
Next, the suspension was filtered and washed with t-BME (200 mL) to
isolate the desired product, 2,9-bis-aminomethylminocycline.
[0167] 2,9-bis-aminomethylminocycline (40 g) was slurried in 200 mL
water/methanol 1/9 and the pH was adjusted to 3 by the dropwise
addition of trifluoroacetic acid. The mixture was heated to
40.degree. C. for 1-2 hours. When HPLC analysis confirmed the
hydrolysis of 2,9-bis-aminomethylminocycline to
9-aminomethylminocycline, the reaction was allowed to return to
room temperature and the pH was adjusted to 7 using triethylamine.
Isopropyl alcohol (200 mL) was added to precipitate out the solid.
The product was filtered and washed with 50 mL IPA followed by 100
mL diethyl ether and dried under reduced pressure toisolate
9-aminomethylminocycline.
9-[(Benzylamino)-methyl]-minocycline dihydrochloride
[0168] To 1.0 mmol (600 mg) of 9-(aminomethyl)-minocycline
dihydrochloride and in 5 mL of dimethylformamide was added 0.2 mmol
(5 mg) of indium trichloride and 1.5 mmol (160 mg) of benzaldehyde
at room temperature. After 30 minutes of shaking, 2 mmol (424 mg)
of sodium triacetoxyborohydride was added and the reaction was
monitored by HPLC. After 1.5 hours, 3 equivalents of triethylamine
and 1 equivalent of sodium triacetoxyborohydride. The reaction was
complete after 3 hours. The solvent was removed in vacuo and the
crude product was purified by preparative HPLC to yield 60 mg of
9-[(benzylamino)-methyl]-minocycline dihydrochloride. LCMS
(MH+)=577.
9-[(2,2,dimethyl-propyl amino)-methyl]-minocycline
dihydrochloride
[0169] 9-dimethylaminominocycline (200 mg, 1 eq.), DMF, and
trimethylacetaldehyde (45 .mu.l, 1 eq.) were combined in 40 mL
flasks and stirred. Triethylamine (150 .mu.L, 3 eq.) was then
added. After stirring at room temperature for several minutes,
NaBH(OAc).sub.3 (175 mg, 2 eq.) and InCl.sub.3 (9 mg, 0.1 eq.) was
added. After one hour, the reactions were clear and red. Liquid
chromatography showed a single product for the reaction. The
reaction was quenched with methanol, the solvent was removed, and
the product was purified using column chromatography.
9-[3,4-(Methylenedioxo)phenyl-ureido]-methylminocycline
dihydrochloride
[0170] To 0.25 mmol (150 mg) of 9-(aminomethyl)-minocycline
dihydrochloride and 2 equivalents of triethylamine in 3 mL of
dimethylformamide was added 0.5 mmol (81.5 mg) of
3,4-(methylenedioxo)phenyl isocyanate at room temperature. Solution
was shaken until reaction was complete (3 hours). Solvent was
removed in vacuo and crude product was purified by preparative HPLC
to yield 66 mg of
9-[3,4-(methylenedioxo)phenyl-ureido]-methylminocycline
dihydrochloride. Yield 41%. LCMS (MH+)=650.
9-[4-(Trifluoromethoxy)phenyl-ureido]-methylminocycline
dihydrochloride
[0171] To 0.25 mmol (150 mg) of 9-(aminomethyl)-minocycline
dihydrochloride and 2 equivalents of triethylamine in 3 mL of
dimethylformamide was added 0.5 mmol (101.5 mg) of
4-(trifluoromethoxy)phenyl isocyanate at room temperature. The
solution was shaken until the reaction was complete (3 hours).
Solvent was removed in vacuo and crude product was purified by
preparative HPLC to yield 68 mg of
9-[4-(trifluoromethoxy)phenyl-ureido]-methylminocycline
dihydrochloride. Yield 39%. LCMS (MH+)=690.
9-(2'-Phenyl-ethyl-1'-amino)-methyl]doxycycline
[0172] Under an N.sub.2 atmosphere, a stirred solution of
9-aminomethyldoxycycline dihydrochloride (1.21 g, 2.21 mmol) in DMF
(10 mL), was treated with InCl.sub.3 (0.076 g, 0.34 mmol) and
phenylacetaldehyde (0.511 mL; 4.4 mmol). HPLC and LCMS monitoring
of the reaction indicated the complete consumption of the starting
material over the course of 12 hours. The products were both the
mono- (major) and bis- (minor) substituted aminodoxycyclines.
Methanol (10 mL) was added to quench the reaction. The reaction
mixture was filtered through a bed of Celite, the celite washed
with methanol (2.times.5 mL), and the combined organic layer was
concentrated to about 7-8 mL and diluted with ether. The resulting
amorphous solid was filtered, washed with ether (6.times.15 mL) and
dried under vacuum to afford a red powder, which was purified by
preparative HPLC. The final product was characterized by HPLC, MS,
and .sup.1H NMR spectroscopic methods. MS (m/z): Theor. 577.24;
Found: 578.17 (M+1).
Example 2
In Vitro Minimum Inhibitory Concentration (MIC) Assay
[0173] The following assay is used to determine the efficacy of
compounds against common bacteria. 2 mg of each compound is
dissolved in 100 .mu.l of DMSO. The solution is then added to
cation-adjusted Mueller Hinton broth (CAMHB), which results in a
final compound concentration of 200 .mu.g per ml. The compound
solutions are diluted to 50 .mu.L volumes, with a test compound
concentration of 0.098 .mu.g/ml. Optical density (OD)
determinations are made from fresh log-phase broth cultures of the
test strains. Dilutions are made to achieve a final cell density of
1.times.10.sup.6 CFU/ml. At OD=1, cell densities for different
genera should be approximately:
TABLE-US-00001 E. coli 1 .times. 10.sup.9 CFU/ml S. aureus 5
.times. 10.sup.8 CFU/ml Enterococcus sp. 2.5 .times. 10.sup.9
CFU/ml
[0174] 50 .mu.l of the cell suspensions are added to each well of
microtiter plates. The final cell density should be approximately
5.times.10.sup.5 CFU/ml. These plates are incubated at 35.degree.
C. in an ambient air incubator for approximately 18 hr. The plates
are read with a microplate reader and are visually inspected when
necessary. The MIC is defined as the lowest concentration of the
compound that inhibits growth. Compounds of the invention indicate
good inhibition of growth.
[0175] In Table 1, compounds which were good inhibitors of growth
of a particular bacteria are indicated with *, compounds which were
very good inhibitors of a particular bacteria are indicated with
**, and compounds with were particularly good inhibitors of a
particular bacteria are indicated with ***.
EQUIVALENTS
[0176] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, numerous
equivalents to the specific procedures described herein. Such
equivalents are considered to be within the scope of the present
invention and are covered by the following claims. The contents of
all references, patents, and patent applications cited throughout
this application are hereby incorporated by reference. The
appropriate components, processes, and methods of those patents,
applications and other documents may be selected for the present
invention and embodiments thereof.
TABLE-US-00002 TABLE 1 ID STRUCTURE S. aureus E. hirae E. coli NZ
##STR00019## ** ** * OA ##STR00020## ** ** * OB ##STR00021## ** **
* OD ##STR00022## *** NT *** OG ##STR00023## ** NT ** OH
##STR00024## *** NT *** OK ##STR00025## ** NT ** OL ##STR00026## *
NT * OM ##STR00027## ** NT * ON ##STR00028## ** NT * OO
##STR00029## ** NT ** OP ##STR00030## ** NT ** OQ ##STR00031## **
NT * OR ##STR00032## * NT * OS ##STR00033## * NT * OT ##STR00034##
* NT * OU ##STR00035## ** NT * OV ##STR00036## ** NT ** PE
##STR00037## NT NT NT PM ##STR00038## NT NT NT PQ ##STR00039## NT
NT NT PR ##STR00040## NT NT NT PS ##STR00041## NT NT NT PT
##STR00042## NT NT NT PV ##STR00043## NT NT NT PW ##STR00044## NT
NT NT
* * * * *