U.S. patent application number 10/652712 was filed with the patent office on 2004-03-11 for 7-substituted fused ring tetracycline compounds.
This patent application is currently assigned to Paratek Pharmaceuticals, Inc.. Invention is credited to McIntyre, Laura, Nelson, Mark L..
Application Number | 20040048835 10/652712 |
Document ID | / |
Family ID | 22756861 |
Filed Date | 2004-03-11 |
United States Patent
Application |
20040048835 |
Kind Code |
A1 |
Nelson, Mark L. ; et
al. |
March 11, 2004 |
7-Substituted fused ring tetracycline compounds
Abstract
7-substituted fused ring tetracycline compounds, methods of
treating tetracycline responsive states, and pharmaceutical
compositions containing the 7-substituted fused ring tetracycline
compounds are described.
Inventors: |
Nelson, Mark L.; (Wellesley,
MA) ; McIntyre, Laura; (Arlington, MA) |
Correspondence
Address: |
LAHIVE & COCKFIELD, LLP.
28 STATE STREET
BOSTON
MA
02109
US
|
Assignee: |
Paratek Pharmaceuticals,
Inc.
Boston
MA
|
Family ID: |
22756861 |
Appl. No.: |
10/652712 |
Filed: |
August 28, 2003 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
10652712 |
Aug 28, 2003 |
|
|
|
09852908 |
May 10, 2001 |
|
|
|
6642270 |
|
|
|
|
60204158 |
May 15, 2000 |
|
|
|
Current U.S.
Class: |
514/152 ;
549/440 |
Current CPC
Class: |
C07C 2603/46 20170501;
C07C 237/26 20130101; C07D 317/60 20130101; A61P 31/00 20180101;
C07D 323/02 20130101; A61P 31/04 20180101 |
Class at
Publication: |
514/152 ;
549/440 |
International
Class: |
A61K 031/65; C07D
317/44 |
Claims
1. A 7-substituted fused ring tetracycline compound of the formula:
12wherein: X is CR.sup.6R.sup.6'; R.sup.4 and R.sup.4' are each
alkyl; R.sup.5 is hydrogen, hydroxyl, or a prodrug moiety; R.sup.6
and R.sup.6' are each independently hydrogen, hydroxyl, alkyl, or
taken together, alkenyl; Y and Y' are each independently optionally
substituted C, N, O, or S; m is 1 or 2; and pharmaceutically
acceptable salts thereof.
2. The compound of claim 1, wherein R.sup.5, R.sup.6 and R.sup.6'
are each hydrogen and R.sup.4 and R.sup.4' are each methyl.
3. The compound of claim 1, wherein Y and Y' are each oxygen.
4. The compound of claim 1, wherein m is 1.
5. The compound of claim 1, wherein said compound is
7-(3',4'-methylene dioxyphenyl)sancycline.
6. A method for treating a tetracycline responsive state in a
mammal, comprising administering to said mammal a 7-substituted
fused ring tetracycline compound of formula (I): 13wherein: X is
CR.sup.6R.sup.6'; R.sup.4 and R.sup.4' are each alkyl; R.sup.5 is
hydrogen, hydroxyl, or a prodrug moiety; R.sup.6 and R.sup.6' are
each independently hydrogen, hydroxyl, alkyl, or taken together,
alkenyl; Y and Y' are each independently optionally substituted C,
N, O, or S; m is 1 or 2; and pharmaceutically acceptable salts
thereof, such that the tetracycline responsive state is
treated.
7. The method of claim 6 wherein R.sup.5, R.sup.6 and R.sup.6' are
each hydrogen and R.sup.4 and R.sup.4' are each methyl.
8. The method of claim 6, wherein Y and Y' are each oxygen.
9. The method of claim 6, wherein m is 1.
10. The method of claim 7, wherein said compound is
7-(3',4'-methylene dioxyphenyl)sancycline.
11. The method of claim 7, wherein said tetracycline responsive
state is a bacterial infection.
12. The method of claim 11, wherein said bacterial infection is
associated with E. coli.
13. The method of claim 11, wherein said bacterial infection is
associated with S. aureus.
14. The method of claim 11, wherein said bacterial infection is
associated with E. faecalis.
15. The method of claim 11, wherein said bacterial infection is
resistant to other tetracycline antibiotics.
16. The method of claim 11, wherein said compound is administered
with a pharmaceutically acceptable carrier.
17. A pharmaceutical composition comprising a therapeutically
effective amount of a compound of claim 1 and a pharmaceutically
acceptable carrier.
18. The pharmaceutical composition of claim 16, wherein said
compound is 7-(3',4'-methylene dioxyphenyl)sancycline.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 09/852,908, filed May 10, 2001, Issuing, which
claims priority to U.S. Provisional Patent Application Serial No.:
60/204,158, filed on May 15, 2000. The entire contents each of the
aforementioned applications are hereby incorporated herein by
reference.
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 tetracycline compounds, without the
ring-attached methyl group present in earlier tetracyclines, was
prepared in 1957 and became publicly available in 1967.
[0003] 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
tetracycline compounds. Examples include U.S. Pat. Nos. 3,957,980;
3,674,859; 2,980,584; 2,990,331; 3,062,717; 3,557,280; 4,018,889;
4,024,272; 4,126,680; 3,454,697; and 3,165,531. These patents are
representative of the range of pharmaceutically active tetracycline
and tetracycline analogue compositions.
[0004] 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 led 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
[0005] The invention pertains to, at least in part, 7-substituted
fused ring tetracycline compounds of the formula: 1
[0006] wherein:
[0007] X is CR.sup.6R.sup.6';
[0008] R.sup.4 and R.sup.4' are each alkyl;
[0009] R.sup.5 is hydrogen, hydroxyl, or a prodrug moiety;
[0010] R.sup.6 and R.sup.6' are each independently hydrogen,
hydroxyl, alkyl, or taken together, alkenyl;
[0011] Y and Y' are each independently optionally substituted C, N,
O, or S;
[0012] m is 1 or 2; and pharmaceutically acceptable salts
thereof.
[0013] In a further embodiment, Y and Y' are each oxygen and m is
1.
[0014] The invention also pertains to a method for treating a
tetracycline responsive state in a mammal, by administering to a
mammal a compound of formula I. In another aspect, the invention
relates to the use of a compound of formula I to treat a
tetracycline responsive state. The invention also pertains to
pharmaceutical compositions comprising a compound of formula I, and
to the use of a compound of formula I in the manufacture of a
medicament to treat a tetracycline responsive state.
DETAILED DESCRIPTION OF THE INVENTION
[0015] The invention includes novel 7-substituted fused ring
tetracycline compounds and methods of using them. In one
embodiment, the invention pertains to 7-substituted fused ring
tetracycline compound of the formula: 2
[0016] wherein:
[0017] R.sup.4 and R.sup.4' are each alkyl;
[0018] R.sup.5 is hydrogen, hydroxyl, or a prodrug moiety;
[0019] R.sup.6 and R.sup.6' are each independently hydrogen,
hydroxyl, alkyl, or taken together, alkenyl;
[0020] Y and Y' are each independently optionally substituted C, N,
O, or S;
[0021] m is 1 or 2; and pharmaceutically acceptable salts
thereof.
[0022] Examples of R.sup.6 and R.sup.6' include methyl, ethyl,
propyl, butyl, pentyl. Together, R.sup.6 and R.sup.6' can be
methylenyl (e.g., methacycline) which may or may not be further
substituted. In a further embodiment, R.sup.5, R.sup.6 and R.sup.6'
are each hydrogen. In another embodiment, R.sup.4 and R.sup.4' are
each lower alkyl, e.g., methyl, ethyl, propyl, butyl, pentyl. In
yet another embodiment, Y and Y' are each oxygen. In one
embodiment, m is 1. In a further embodiment, the compound is
7-3',4'-methylenedioxyphenyl sancycline.
[0023] In a further embodiment, Y and Y' are substituted or
unsubstituted such that the compound can perform its intended
function. For example, if Y or Y' is C or N, the substituent can be
hydrogen, alkyl (e.g., methyl, ethyl, propyl, etc.), halogen,
hydroxy, or any other substituent which either allows the compound
to perform its function or enhances its ability to do so.
Furthermore, the 7-substituent may also be substituted at any of
the other positions of either ring. Examples of possible
substituents include alkyl, alkenyl, alkynyl, halogen, hydroxyl,
alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkylamino,
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.
In an embodiment, m is one and Y and Y' are oxygen. In a further
embodiment, the 7-substituent of the tetracycline compound is
methylenedioxyphenyl.
[0024] The term "tetracycline compound" includes compounds with a
similar ring structure to tetracycline, such as those included in
formula I. Some examples of tetracycline compounds which can be
modified to include a substituent at position 7 include
tetracycline, oxytetracycline, methacycline, sancycline, and
doxycycline; however, other derivatives and analogues comprising a
similar ring structure are also included. Table I depicts
tetracycline and several known tetracycline derivatives.
1TABLE I 3 4 5 6 7 8 9
[0025] The term "7-substituted fused ring tetracycline compounds"
includes tetracycline compounds with a fused ring at the 7
position. In an embodiment, the substituted tetracycline compound
is substituted tetracycline (e.g., wherein R.sup.4 and R.sup.4' are
methyl, R.sup.5 is hydrogen, R.sup.6 is methyl and R.sup.6' is
hydroxyl); substituted doxycycline (e.g., wherein R.sup.4 and
R.sup.4' are methyl, R.sup.5 is hydroxyl R.sup.6 is methyl and
R.sup.6' is hydrogen); or substituted sancycline (wherein R.sup.4
and R.sup.4' are methyl; R.sup.5 is hydrogen and R.sup.6 and
R.sup.6' are hydrogen atoms). In another embodiment, the compound
is a derivative of tetracycline, minocycline, sancycline,
doxycycline, chlortetracycline, oxytetracycline, demeclocycline, or
methacycline.
[0026] The term "fused ring" includes moieties of the formula:
10
[0027] wherein m is 1 or 2, and Y and Y' are each independently
selected from the group consisting of substituted or unsubstituted
O, N, S, or C. Y and Y' are substituted or unsubstituted such that
the compound can perform its intended function. For example, if Y
or Y' is C or N, the substituent can be, for example, hydrogen,
alkyl (e.g., methyl, ethyl, propyl, etc.), halogen, hydroxy, or
another substituent which allows the compound to perform its
intended function. Furthermore, the fused ring may also be
substituted at any of the other positions of either ring. Examples
of possible substituents include alkyl, 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.
In an embodiment, m is one and Y and Y' are oxygen. In a further
embodiment, the fused ring is methylenedioxyphenyl.
[0028] In one embodiment, the 7-substituted fused ring tetracycline
compound is 7-(3',4'-methylene dioxyphenyl)sancycline.
[0029] The 7-substituted fused ring compounds of the invention can
be synthesized by methods known in the art and/or as described
herein. In Scheme 1, a general synthetic scheme is outlined using a
Suzuki coupling of a boronic acid with an iodo tetracycline
compound. Although the reaction is shown for sancycline, a similar
procedure can be used for other tetracycline compounds.
Furthermore, other aryl coupling reactions known in the art may
also be used. 11
[0030] As shown in Scheme 1, an iodosancycline compound can be
synthesized from unsubstituted sancycline by treating it with at
least one equivalent N-iodosuccinimide (NIS) under acidic
conditions. When sancycline was treated with NIS in trifluoroacetic
acid, the reaction was carried out initially at 0.degree. C.,
before being warmed to room temperature for five hours. The
reaction is then quenched, and the resulting 7-iodosancycline can
then be purified using standard techniques known in the art. The
7-iodosancycline can then be further reacted with a boronic acid,
as shown in Scheme 1. 7-iodosancycline, a palladium catalyst (such
as Pd(OAc).sub.2), is dissolved in a solvent and treated with
aqueous sodium carbonate, and the boronic acid. The resulting
compound can then be purified using techniques known in the art
such as preparative HPLC and characterized. The synthesis of the
compounds of the invention are described in more detail in Example
1.
[0031] 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 comprise 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.
[0032] 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, alkyl, alkenyl, alkynyl, halogen, hydroxyl,
alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkylamino,
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 "aralkyl" 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.
[0033] The term "aryl" includes groups with aromaticity, including
5- and 6-membered single-ring aromatic groups that may include from
zero to four heteroatoms as well as multicyclic systems with at
least one aromatic ring. Examples of aryl groups include benzene,
phenyl, pyrrole, furan, thiophene, thiazole, isothiazole,
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, alkylaminocarbonyl,
aralkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl,
arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylthiocarbonyl, phosphate, phosphonato,
phosphinato, cyano, amino (including alkylamino, 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 multicyclic system (e.g., tetralin,
methylenedioxyphenyl).
[0034] 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.
[0035] For example, the term "alkenyl" includes straight-chain
alkenyl groups (e.g., ethenyl, 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.
[0036] 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 alkylamino,
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.
[0037] 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.
[0038] 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.
[0039] 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, alkylaaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkylamino,
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.
[0040] 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.
[0041] Is 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 alkylamino, 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.
[0042] 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.
[0043] 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.
[0044] 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.
[0045] 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 alkylamino,
dialkylamino, arylaamino, 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.
[0046] The term "amine" or "amino" includes compounds where a
nitrogen atom is covalently bonded to at least one carbon or
heteroatom. The term "alkylamino" includes groups and compounds
wherein the nitrogen is bound to at least one additional alkyl
group. The term "dialkylamino" 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.
[0047] The term "amide" or "aminocarboxy" 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
"alkaminocarboxy" groups which include alkyl, alkenyl, or alkynyl
groups bound to an amino group bound to a carboxy group. It
includes arylaminocarboxy 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 "alkylaminocarboxy,"
"alkenylaminocarboxy," "alkynylaminocarboxy," and
"arylaminocarboxy" include moieties wherein alkyl, alkenyl, alkynyl
and aryl moieties, respectively, are bound to a nitrogen atom which
is in turn bound to the carbon of a carbonyl group.
[0048] 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.
[0049] The term "thiocarbonyl" or "thiocarboxy" includes compounds
and moieties which contain a carbon connected with a double bond to
a sulfur atom.
[0050] 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.
[0051] 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.
[0052] 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.
[0053] The term "hydroxy" or "hydroxyl" includes groups with an
--OH or --O.sup.-.
[0054] 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.
[0055] 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. 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,
alkylaminocarbonyl, aralkylaminocarbonyl, alkenylaminocarbonyl,
alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl,
aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato,
phosphinato, cyano, amino (including alkylamino, 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.
[0056] The term "heteroatom" includes atoms of any element other
than carbon or hydrogen. Examples of heteroatoms include nitrogen,
oxygen, sulfur and phosphorus.
[0057] 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.
[0058] Prodrugs are compounds which are converted in vivo to active
forms (see, e.g., R. B. Silverman, 1992, "The Organic Chemistry of
Drug Design and Drug Action", Academic Press, Chp. 8). Prodrugs can
be used to alter the biodistribution (e.g., to allow compounds
which would not typically enter the reactive site of the protease)
or the pharmacokinetics for a particular compound. For example, a
hydroxyl group, can be esterified, e.g., with a carboxylic acid
group to yield an ester. When the ester is administered to a
subject, the ester is cleaved, enzymatically or non-enzymatically,
reductively or hydrolytically, to reveal the hydroxyl group.
[0059] The term "prodrug moiety" includes moieties which can be
metabolized in vivo to a hydroxyl group and moieties which may
advantageously remain esterified in vivo. Preferably, the prodrugs
moieties are metabolized in vivo by 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. (I 977) "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 in its free acid form or hydroxyl
with a suitable esterifying 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, dilower 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.
[0060] The invention also features a method for treating a
tetracycline compound responsive state in a subject, by
administering to the subject a 7-substituted fused ring
tetracycline compound of the invention. Preferably, an effective
amount of the tetracycline compound is administered. In an
embodiment, the compound is 7-(3',4'-methylene
dioxyphenyl)sancycline.
[0061] The language "tetracycline compound responsive state"
includes states which can be treated, prevented, or otherwise
ameliorated by the administration of a tetracycline compound of the
invention. Tetracycline compound responsive states include
bacterial infections (including those which are resistant to other
tetracycline compounds), cancer, diabetes, and other states for
which tetracycline compounds have been found to be active (see, for
example,
[0062] U.S. Pat. Nos. 5,789,395; 5,834,450; and 5,532,227).
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)).
[0063] 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).
[0064] The tetracycline compounds 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.
[0065] 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 tetracycline compound. For example, the
choice of the tetracycline 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.
[0066] 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
tetracycline compounds to a subject. The subject can be either a
plant or, advantageously, an animal, e.g., a mammal, e.g., a
human.
[0067] In the therapeutic methods of the invention, one or more
tetracycline 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.
[0068] In one embodiment, the pharmaceutical composition comprises
a 7-substituted fused-ring tetracycline compound of the invention,
e.g., of formula I. In an embodiment, the compound is
7-(3',4'-methylene dioxyphenyl)sancycline.
[0069] The language "pharmaceutically acceptable carrier" includes
substances capable of being coadministered with the tetracycline
compound(s), and which allow both to perform their intended
function, e.g., treat or prevent a tetracycline compound 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.
[0070] The tetracycline 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
tetracycline 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-na- phthoate)] 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 tetracycline 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 tetracycline 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.
[0071] The preparation of other tetracycline 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.
[0072] The tetracycline 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 tetracycline
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 tetracycline 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 tetracycline 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 tetracycline compound of the invention may be mixed with an
alkoxide of the desired metal and the solution subsequently
evaporated to dryness.
[0073] The preparation of other tetracycline 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.
[0074] The tetracycline 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.
[0075] The pharmaceutical compositions of the invention may be
administered alone or in combination with other known compositions
for treating tetracycline responsive states in 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.
[0076] 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.
[0077] 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.
[0078] 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.
[0079] 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.
[0080] 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.
[0081] 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.
[0082] 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.
[0083] 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.
[0084] It will also be understood that normal, conventionally known
precautions will be taken regarding the administration of
tetracyclines 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.
[0085] Furthermore, the invention also pertains to the use of a
tetracycline compound of formula I, for the preparation of a
medicament. In one embodiment, the tetracycline compound is
7-3',4'-methylenedioxyphe- nyl sancycline. The medicament may
include a pharmaceutically acceptable carrier and the tetracycline
compound is an effective amount, e.g., an effective amount to treat
a tetracycline responsive state.
[0086] In yet another embodiment, the invention also pertains to
the use of a tetracycline compound of formula 1 to treat a
tetracycline responsive state, e.g., in a subject, e.g., a mammal,
e.g., a human.
EXEMPLIFICATION OF THE INVENTION
[0087] 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 7-(3',4'-Methylene dioxyphenyl)sancycline
[0088] 7-Iodosancycline
[0089] One gram of sancycline was dissolved in 25 mL of
trifluoroacetic acid that was cooled to 0.degree. C. (on ice). 1.2
equivalents of N-iodosuccinimide (NIS) was added to the reaction
mixture and reacted for forty minutes. The reaction was removed
from the ice bath and was allowed to react at room temperature for
an additional five hours. The mixture was then analyzed by HPLC and
TLC, was driven to completion by the stepwise addition of NIS.
After completion of the reaction, the TFA was removed in vacuo and
3 mL of MeOH was added to dissolve the residue. The methanolic
solution was then added slowly to a rapidly stirring solution of
diethyl ether to form a greenish brown precipitate. The 7-iodo
isomer of sancycline is purified by treating the 7-iodo product
with activated charcoal, filtering through Celite, and subsequent
removal of the solvent in vacuo to produce the 7-isomer compound as
a pure yellow solid in 75% yield.
[0090] MS (M+H) (formic acid solvent): 541.3;
[0091] Rt: Hypersil C18 BDS Column, 11.73;
[0092] .sup.1H NMR: 300 MHz (Methanol d.sub.4, TMS):
.delta.7.87-7.90 (d, 1H); 6.66-6.69 (d, 1H); 4.06;
[0093] (s, 1H); 2.98 (s, 6H); 2.42 (m, 1H); 2.19 (m, 1H); 1.62 (m,
4H); 0.99 (m, 2H).
[0094] 7-(3 ',4'-Methylene dioxyphenyl)sancycline
[0095] 200 mg of 7-iodosancycline (0.3 mM), 8.4 mg of Pd(OAc).sub.2
(10% mole equiv.) and MeOH (5 mL) were added to a flask and the
system was purged with dry argon while heating externally.
Na.sub.2CO.sub.3 (117 mg, 3 eq.) dissolved in water (2 mL purged
with argon)were added to the reaction flask and purged with argon.
The boronic acid (3',4'-methylenedioxyphenyl boronic acid) (123 mg,
2 eq.) was dissolved in MeOH (5 mL) and the system was purged with
argon for 5 minutes. This solution was then added via syringe to
the flask and allowed to react for 1-2 hours. The reaction was then
stopped and the solvent was removed in vacuo to produce the crude
product. The mixture was purified by preparative HPLC column
chromatography using divinylbenzene as a solid phase and a binary
solvent system of trifluoroacetic acid (0.1%) and acetonitrile over
a gradient of 0% to 100% ACN over 20 minutes. The compound peak
eluted at 13.7 minutes. The fractions were removed and the solvent
was removed in vacuo and the product isolated as the HCL salt by
bubbling HCl gas (anhydrous) through a methanolic solution for 3
minutes and subsequent removal of the solvent. The compound was
dried in vacuo to yield the compound as a bright yellow solid in
43% yield.
[0096] MS: M+H (formic acid solvent): 535.2;
[0097] Rt:, divinyl benzene solid-phase 13.7 minutes;
[0098] .sup.1H NMR: 300 MHz (methanol d.sub.4, TMS)
.delta.7.47-7.50 (d, 1H), 6.97 (m, 2H), 6.81 (m, 2H), 6.07;
[0099] (s, 2H), 4.14 (s, 1H), 2.99 (s, 6H), 2.59 (m, 1H), 2.12 (m,
1H), 1.64 (m, 1H)
Example 2
In Vitro Minimum Inhibitory Concentration (MIC) Assay
[0100] The following assay was used to determine the efficacy of
tetracycline compounds against common bacteria. 2 mg of each
compound was dissolved in 100 .mu.l of DMSO. The solution was then
added to cation-adjusted Mueller Hinton broth (CAMHB), which
resulted in a final compound concentration of 200 .mu.g per ml. The
tetracycline compound solutions were diluted to 50 .mu.L volumes,
with a test compound concentration of 0.098 .mu.g/ml. Optical
density (OD) determinations were made from fresh log-phase broth
cultures of the test strains. Dilutions were made to achieve a
final cell density of 1.times.10.sup.6 CFU/ml. At OD=1, cell
densities for different genera are approximately:
2 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
[0101] 50 .mu.l of the cell suspensions were added to each well of
the microtiter plates. The final cell density was approximately
5.times.10.sup.5 CFU/ml. These plates were incubated at 35.degree.
C. in an ambient air incubator for approximately 18 hr. The plates
were read with a microplate reader and were visually inspected when
necessary. The MIC is defined as the lowest concentration of the
tetracycline compound that inhibits growth.
[0102] Table 2 shows the relative MIC values for 7-(3',4'-methylene
dioxyphenyl)sancycline. For the table, * indicates good inhibition
of growth, ** indicates very good inhibition of growth, and ***
indicates exemplary inhibition of growth.
3TABLE 2 INHI- INHI- ORGANISM BITION ORGANISM BITION E. coli Dl-299
* S.aureus 4250 ** E. coli D1-209 * S. aureus ATCC 29213 *** E.
ccli pHCM1 ** S. aureus ATCC 13709 *** S. aureus 12715 *** S.
pnuemoniae ATCC 49619 *** E. faec pMV158 *** S. pnuemoniae ATCC
157E *** E. faec pAM211 ** E. hirae ATCC 9790 *** E. coli ML308-225
* H. influenzae ATCC 49247 *** S. aureus RN450 *** M. catarrhalis
ATCC 23246 ** E. faecalis ATCC *** E. faecalis ATCC 9790 ** 9790
ATCC 29212 *** mrsa 5 ***
EQUIVALENTS
[0103] 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, issued patents, and published 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.
* * * * *