Note: Descriptions are shown in the official language in which they were submitted.
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SUBSTITUTED TETRACYCLINE COMPOUNDS
This application is a divisional application of co-pending Application Serial
No. CA 2,531,728, filed June 25, 2004.
Background of the Invention
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; 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
tetracycline
compounds. Examples include U.S. Patent 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.
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
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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:
In one embodiment, the invention pertains to a 7,9-substituted tetracycline
compound of Formula I:
R7 Rs R4
x ow
R9
12
OR
R0 O R" (j)
wherein:
Xis CHC(R13Y'Y), CR6'R6, S, NR6, or 0;
R2, R2', R4', and e' are 'each independently hydrogen, alkyl, alkenyl,
alkynyl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic,
heteroaromatic or a prodrug moiety;
R4 is I~R4R4", alkyl, alkenyl, alkynyl, hydroxyl, halogen, or hydrogen;
RZ*, R3, R' , Rn and Rte are each hydrogen or a pro-drug moiety;
R5 is hydroxyl, hydrogen, thiol, alkanoyl, aroyl, alkaroyl, aryl,
heteroaromatic,
alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,
alkylamino,
arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy,
R6 and R6' are each independently hydrogen, methylene, absent, hydroxyl,
halogen, thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy, alkylthio,
alkylsulfinyl,
alkylsulfonyl, alkylamino, or an arylalkyl;
R' ethyl, perhalogenated alkenyl, substituted pyridinyl, pyrazinyl, furanyl,
or
pyrazolyl;
R$ is hydrogen, hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl,
alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;
R9 -CH2NR9R9b;
R9a and R91' are each independently hydrogen, alkyl, alkenyl or linked to form
a
heterocycle;
R13 is hydrogen, hydroxy, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,
alkylsulfinyl,
alkylsulfonyl, alkylamino, or an arylalkyl; and
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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.
In another embodiment, the invention pertains to a 9-substituted tetracycline
compound of formula II:
R7 RS R4
x OR?
I NIORr
R9
ORja
R10 0 OR" 0 0 (II)
wherein:
Xis CHC(R13Y'Y), CR6'R6, S, NR6, or 0;
R2, R4', R4", R7' and R7" are each hydrogen, alkyl, alketiyl, alkynyl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic,
heteroaromatic or a prodrug moiety;
R4 is NR4'R4", alkyl, alkenyl, alkynyl, aryl, hydroxyl, halogen, or hydrogen;
R2', R3, 0, R" and RI2 are each hydrogen or a pro-drug moiety;
R5 is hydroxyl, hydrogen, thiol, alkanoyl, aroyl, alkaroyl, aryl,
heteroaromatic,
alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,
alkylamino,
arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy,
R6 and R6' are independently hydrogen, methylene, absent, hydroxyl, halogen,
thiol, alkyl, alkenyl, alk}myl, aryl, alkoxy, alkylthio, alkylsulfinyl,
alkylsulfonyl,
alkylamino, or an arylalkyl;
R7 is NR7'R7", alkyl, alkenyl, alkynyl, aryl, hydroxyl, halogen, or hydrogen;
s i
R is hydrogen, hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl,
alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;
R9 is -CH2NR fte, or linked with R10 to form a furanyl ring;
R9a is hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl,
alkylsulfonyl, alkylamino, arylalkyl, aryl, heterocyclic,or heteroaromatic;
e is hydrogen or alkyl;
R$ is hydrogen, hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl,
alkoxy,,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;
RP is hydrogen, hydroxy, alkyl, alkeny1, 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,
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alkylamino, or an arylalkyl, and pharmaceutically acceptable salts, esters and
prodrugs
thereof.
In another embodiment, the invention pertains to 7-substituted tetracycline
compounds of formula III:
R7 RS R
Rs X R3
I I NR2Rr
_
R9 =
ORS'
R10 O R~~ 0 0 lhl
Wherein:
X is CHC(Ri3Y'Y), CR6'R6, C=CR6'R6, S, NR6, or 0;
R2, R2', R4', and R4" are each independently hydrogen, alkyl, alkenyl,
alkynyl,
alkoxy, aikyltbio, alkylsulfonyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic,
heteroaromatic or a prodrug moiety;
R4 is NR4'R4", alkyl, alkenyl, alkynyl, aryl, hydroxyl, halogen, or hydrogen;
R2', R3, R10, R1' and R12 are each hydrogen or a pro-drug moiety;
RS is hydroxyl, hydrogen, thiol, alkanoyl, aroyl, alkaroyl, aryl,
heteroaromatic,
alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfnyl, alkylsulfonyl,
alkylamino,
arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy,
R6 and R6' are each independently hydrogen, methylene, absent, hydroxyl,
halogen, thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy, alkylthio,
alkylsulfinyl,
alkylsulfonyl, alkylamino, or an arylalkyl;
R7 is substituted or unsubstituted pyrazolyl, furanyl, thiophenyl, or
thiazolyl;
R8 is hydrogen, hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl,
alkoxy,
alkylthio, alkylsulfnyl, alkylsulfonyl, alkylamino, or an arylalkyl;
R4 is hydrogen;
R13 is hydrogen, hydroxy, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,
alkylsulfnyl,
alkylsulfonyl, alkylamino, or an arylalkyl; and
Y' and Y are each independently hydrogen, halogen, hydroxyl, cyano,
sulfhydryl,
amino, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulnyl, alkylsulfonyl,
alkylamino, or an arylalkyl, and pharmaceutically acceptable salts thereof
In another embodiment, the invention pertains to 8-substituted tetracycline
compound of formula IV:
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R7 RS R
R / OR10 O ORII O O
wherein:
X is CHC(R13Y'Y), CR6'R6, S, NR6, or 0;
R2, e, R4'; R7 and RT' are each hydrogen, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfnyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic,
heteroaromatic or a prodrug moiety;
R4 is NR4'R4", alkyl, alkenyl, alkynyl, aryl, hydroxyl, halogen, or hydrogen;
R2', R3, R10, R" and Rig are each hydrogen or a pro-drug moiety,
R5 is hydroxyl, hydrogen, thiol, alkanoyl, aroyl, alkaroyl, aryl,
heteroaromatic,
alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,
alkylamino,
arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy,
R6 and R6' are independently hydrogen, methylene, absent, hydroxyl, halogen,
thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl,
alkylsulfonyl,
alkylamino, or an arylalkyl;
R7 is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl, alkynyl,
aryl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, arylalkyl, amino,
arylalkenyl, arylalkynyl,
acyl, aminoalkyl, heterocyclic, thionitroso, or-(CH2)as (NR'`)o.iC(w')wR7a;
R8 is an aminomethyl substituted phenyl or substituted pyridinyl;
R9 is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl, alkynyl,
aryl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, arylalkyl, amino,
arylalkenyl, arylalkynyl,
acyl, aminoalkyl, heterocyclic, thionitroso, or-(CH2)a3NR9C(=Z')ZR9a;
R7a, Rye, e, R7d, R7e, e, R9a, R91', R9c, R9d, R9e, and R81 are each
independently
absent, hydrogen, acyl, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,
alkylsulfnyl,
alkylsulfonyl, alkylamino, arylalkyl, aryl, heterocyclic, heteroaromatic or a
prodrug
moiety;
W is CRrdRRe, S, 0 or NRh;
W' is 0, W1. or S;
Z is CR9dR9e, S, 0 or NR9b;
Z'is 0,NR9,or S;
R13 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,
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alkylamino, or an arylalkyl, and pharmaceutically acceptable salts, esters and
prodrugs
thereof.
In one embodiment, a 13-substituted tetracycline compound is of formula V:
Rt3
R7 I R5_ R4
Rs OW
R9
OR12
(V)
OR10 0 OR11 0 0
wherein:
R2, R4', R4", R7' and RT' are each hydrogen, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfnyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic,
heteroaromatic or a prodrug moiety,
RR is NR4'R4õ, alkyl, alkenyl, alkynyl, aryl, hydroxyl, halogen, or hydrogen,
R2', R3, R10, R" and R'2 are each hydrogen or a pro-drug moiety;
R5 is hydroxyl, hydrogen, thiol, alkanoyl, amyl, alkaroyl, aryl,
heteroaromatic,
alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,
alkylamino,
arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy,
R7 is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl, alkynyl,
aryl,
alkoxy, alkylthio, alkylsulfnyl, alkylsulfonyl, arylalkyl, amino, arylalkenyl,
arylalkynyl,
acyl, aminoalkyl, heterocyclic, thionitroso, or -(CH2)0-3 (NR7`)o-iC(=W')WRia;
R8 is substituted phenyl or substituted pyridinyl;
R9 is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl, alkynyl,
aryl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, arylalkyl, amino,
arylalkenyl, arylalkenyl,
acyl, aminoalkyl, heterocyclic, thionitroso, or -(CH2)o-3NR9oC(=Z')ZR9a;
R7a, RT', R7c, R7d, R7e, R7 , R9a, R9b, R9o, R9d, R9, and R8 are each
independently
absent, hydrogen, acyl, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,
alkylsulfnyl,
alkylsulfonyl, alkylamino, arylalkyl, aryl, heterocyclic, heteroaromatic or a
prodrug
moiety;
W is CR7dR7e, S, 0 or NR7
W' is 0,NR7,orS;
R13 is 4-alkyl substituted phenyl, and pharmaceutically acceptable salts,
esters
and prodrugs thereof.
In another f irther embodiment, the invention pertains, at least in part, to
methods
for treating subjects for tetracycline responsive states by administering to
them an
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effective amount of a tetracycline compound of the invention, e.g., a compound
of
formula I, II, III, IV, V, or a tetracycline compound otherwise described
herein.
Detailed Description of the Invention:
The present invention pertains, at least in part, to novel substituted
tetracycline
compounds. These tetracycline 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 tetracycline compounds in general, such
as
blocking tetracycline efflux and modulation of gene expression.
The term "tetracycline compound" includes many compounds with a similar ring
structure to tetracycline. Examples of tetracycline compounds include:
chlortetracycline, oxytetracycline, demeclocycline, methacycline, sancycline,
chelocardin, rolitetracycline, lymecycline, apicycline; clomocycline,
guamecycline,
meglucycline, mepylcycline, penimepicycline, pipacycline, etamocycline,
penimocycline, etc. Other derivatives and analogues comprising a similar four
ring
structure are also included (See Rogalski, "Chemical Modifications of
Tetracyclines."
Table 1 depicts tetracycline and several known other tetracycline derivatives.
Table 1
H OH H H h 2h
i x
OH
O O OH O
Oxybelracycline Demecloc line Minocycline
co
H 'h p nc +h
H x
0 0 0
M0" 0 ON 0
ethacycline Doxycycliae Chlortetra cline
F{,C H N(+Mh N"). H, Pfth ii %-J~
o Sancycline Chelocardm
Tetracycline
Other tetracycline compounds which maybe modified using the methods of the
invention include, but are not limited to, 6-demethyl-6-deoxy-4-
dedimethylaminotetracycline; tetracyclino-pyrazole; 7-chloro-4-
dedimethylaminotetracycline; 4-hydroxy-4-dedimethylaminotetracycline; 12a-
deoxy-4-
dedimethylaminotetracycline; 5-hydroxy-6a-deoxy-4-ded methylaminotetracycline;
4-
dedimethylamino-l2a-deoxyanhydrotetracycline; 7-dimethylamino-6-demethyl-6-
deoxy-4-dedimethylaminotetracycline; tetracyclinonitrile; 4-oxo-4-
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dedimethylaminotetracycline 4,6-hemiketal; 4-oxo-11a C1-4-
dedimethylaminotetracycline-4,6-hemiketal; 5a,6-anhydro-4-hydrazon-4-
dedimethylamino tetracycline; 4-hydroxyimino-4-dedimethylamino tetracyclines;
4-
hydroxyimino-4-dedimethylamino 5a,6-anhydrotetracyclines; 4-amino-4-
dedimethylamino-5a, 6 anhydrotetracycline; 4-methylamino-4-dedimethylamino
tetracycline; 4-hydrazono-1la-chloro-6-deoxy-6-demethyl-6-methylene-4-
dedimethylamino tetracycline; tetracycline quaternary ammonium compounds;
anhydrotetracycline betaines; 4-hydroxy-6-methyl pretetramides; 4-keto
tetracyclines; 5-
keto tetracyclines; 5a, 1l a dehydro tetracyclines; 11 a CI-6, 12 hemiketal
tetracyclines;
11a CI-6-methylene tetracyclines; 6, 13 diol tetracyclines; 6-
benzylthiomethylene
tetracyclines; 7, 11 a -dichloro-6-fluoro-methyl-6-deoxy tetracyclines; 6-
fluoro ((x)-6-
denlethyl-6-deoxy tetracyclines; 6-fluoro ((3)-6-demethyl-6-deoxy
tetracyclines;6-a
acetoxy-6-demethyl tetracyclines; 6-P acetoxy-6-demethyl tetracyclines; 7, 13-
epithiotetracyclines; oxytetracyclines; pyrazolotetracyclines; 1la halogens of
tetracyclines; 12a formyl and other esters of tetracyclines; 5, 12a esters of
tetracyclines;
10, 12a- diesters of tetracyclines; isotetracycline; 12-a-deoxyanhydro
tetracyclines; 6-
domethyl-12a-deoxy-7-chloroanhydrotetracyclines; B-nortetracyclines; 7-methoxy-
6-
demethyl-6-deoxytetracyclines; 6-demethyl-6-deoxy-5aepitetracyclines; 8-
hydroxy-6-
demethyl-6-deoxy tetracyclines; monardene; chromocycline; 5a methyl-6-demethyl-
6-
deoxy tetracyclines; 6-oxa tetracyclines, and 6 thia tetracyclines.
1. 7,9-Substituted Tetracycline Compounds
The invention also pertains, at least in part to 7,9-substituted tetracycline
compounds.
The term "7,9-substituted tetracycline compounds" includes tetracycline
compounds with substitution at the 7 and 9- positions. In one embodiment, the
substitution at the 7- and 9- positions enhances the ability of the
tetracycline compound
to perform its intended function, e.g., treat tetracycline responsive states.
In an
embodiment, the 7,9-substituted tetracycline compound is 7,9-substituted
tetracycline
(e.g., wherein R4 is NR4'R4"; R4' and R4" are methyl, R5 is hydrogen and X is
CR6R6',
wherein R6 is methyl and R6' is hydroxy); 7,9-substituted doxycycline (e.g.,
wherein R4
is NR4'R4"; R4' and R4" are methyl, R5 is hydroxyl and X is CR6R6', wherein R6
is methyl
and R6' is hydrogen); or 7,9- substituted sancycline (wherein R4 is NR4'R4";
R4' and R4..
are methyl; R5 is hydrogen and X is CR6R6' wherein R6 and R6' are hydrogen
atoms. In
an embodiment, the substitution at the 7 position of the 7, 9-substituted
tetracycline
compound is not chlorine or trimethylamino. In one embodiment, R4 is hydrogen.
In one embodiment, the invention pertains to 7,9-substituted tetracycline
compounds of Formula I:
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R7 RS e
s
R X ORS
R9
OR~Z
Rio O OR"' O O
wherein:
X is CHC(R13Y'Y), CR6R6, S, NR6, or 0;
e, R , e, and R are each independently hydrogen, alkyl, alkenyl, alkynyl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic,
heteroaromatic or a prodrug moiety;
R4 is NR4'R4", alkyl, alkenyl, alkynyl, hydroxyl, halogen, or hydrogen;
R2', R3, R' , R" and R12 are each hydrogen or a pro-drug moiety;
R5 is hydroxyl, hydrogen, thiol, alkanoyl, amyl, alkaroyl, aryl,
heteroaromatic,
alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,
alkylamino,
arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy;
R6 and R6' are each independently hydrogen, methylene, absent, hydroxyl,
halogen, thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy, alkylthio,
alkylsulfinyl,
alkylsulfonyl, alkylamino, or an arylalkyl;
R7 is ethyl, perhalogenated alkenyl, substituted pyridinyl, pyx"azinyl,
furanyl, or
pyrazolyly
R$ is hydrogen, hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl,
alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;
R9 is -CH2NR9aR9b; `
R9a and R9b are each independently hydrogen, alkyl, alkenyl or linked to form
a
heterocycle;
R13 is hydrogen, hydroxy, alkyl, alkenyl, alkenyl, alkoxy, alkylthio,
alkylsufnyl,
alkylsulfonyl, alkylamino, or an arylalkyl; and
Y' and Y are each independently hydrogen, halogen, hydroxyl, cyano,
sulfhydryl,
amino, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulilnyl,
alkylsulfonyl,
alkylamino, or an arylalkyl, and pharmaceutically acceptable salts thereof,
provided that
R7 and R9 are not both unsubstituted phenyl.
In a further embodiment, X is CR6R6'; R2, R2', e, R6', e, Rio, R", and R12 are
each hydrogen; R4 is NR4'R4"; R4' and R4" are lower alkyl; and R5 is hydroxy
or
hydrogen. In another further embodiment, R4' and R4" are each methyl and R5 is
hydrogen.
-9-
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In an embodiment, R7 is ethyl and R9a is alkyl and R9b is alkenyl. In another
embodiment, R7 is substituted pyrazinyl Examples of possible substituents
include
halogens, such as fluorine. In another embodiment, R9a is alkyl and R9b is
alkenyl. In
another further embodiment, RR' and R9b are linked to form a heterocycle. In a
further
embodiment, the linked heterocycle is substituted piperidinyl. In a further
embodiment,
the piperdinyl is substituted with one or more fluorines or halogenated alkyl
groups, e.g.,
at the 2, 3, 4, or 5 position. In another embodiment, the R9 moiety is
(4'trifluoromethyl-
piperdin-1-yl) methyl, (4', 4'-difluoro-piperdin-l-yl) methyl, or (4'-
fluoropiperdin-l-yl)
methyl.
In another embodiment, R9a is hydrogen and R9b is alkyl. Other examples of
compounds include those wherein R7 is furanyl, and R9a is hydrogen or alkyl
and R9b is
alkenyl, e.g., 1, 2, 2-trifluoroethenyl.
In another embodiment, R9a is hydrogen or alkyl and R9b is alkenyl. In another
embodiment, R7 is pyrazolyl and R9a is hydrogen or alkyl and R9b is alkenyl or
alkyl.
In a further embodiment, the invention pertains to tetracycline compounds
selected from the group consisting of-
cH, H,C .CHa
H_ kl
0NH, H Hi-tea H ~H
OOV~ _
0 H2
a
OH, OH O O
OH 0 OtP
F HaC- -CH, H,C.N.CH2
Hac CHa H H -
x {~_ _ - OH - ` OH
O' H _ NHs N
ONO o O OH 0HH - O CH3 OH O O
F F H HiH -tea N ..CH, F F H.C'N.CH,
D OH H H H H H OH
H OH
NH2
NN N W6
F OH O OH O OH O
^ S'~ 4FF
- Hs V HsU H H
OH M42
OH 0 O H
F - ' NH,
F F H O O
rc lot
NCF'H H C~ NH, CF{, Oti i. N=C OH a H N'~+
1_ OH
- OH F _ NHz
Ha 2 NH2 I / \ NH, OH O
ON O OH O OFf
CH3 Ha-dii CHt H,C.N.CH, a fa
F \ H_ H - OH F \ H H tNJ H,C N.CH,
F~N / a ( H2 N NH, F`^ OH
OH O OH O OH O OH O N_ NH,
OH O
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H=N H C pH\N'C~ F F or
oH ~
H C C I\ I NHs HC.N.CH, H ~H OH
H H ~ - -
H,c off o off o o _ off I M4
HZN I I H2 OH o o& o
off o otP o
H,C.N_CH,
I / H,C-N,OH,
H L1 OH
H,N I / _ I NH,
OH O 0
and pharmaceutically acceptable salts, esters, and prodrugs thereof.
2. 9-Substituted Tetracycline Compounds
In another embodiment, the invention pertains to 9-substituted tetracycline
compounds.
The term "9-substituted tetracycline compounds" includes tetracycline
compounds with substitution at the 9 position. In one embodiment, the
substitution at
the 9- position enhances the ability of the tetracycline compound to perform
its intended
function, e.g., treat tetracycline responsive states. In an embodiment, the 9-
substituted
10. tetracycline compound is 9-substituted tetracycline (e.g., wherein Ra is
NR4'R`r', R4' and
Ra,. are methyl, RS is hydrogen and X is CR6R6', wherein R6 is methyl and R6'
is
hydroxy, and R7 is hydrogen); ; 9-substituted doxycycline i a' R 4
xycycline (e.g., wherein e is NR ,
R4' and R4' are methyl, R5 is hydroxyl and X is CR6R6', wherein R6 is methyl
and R6' is
hydrogen, and R7 is hydrogen); 9- substituted minocycline (wherein Ra is
NR4'R4", Ra'
and e are methyl; RS is hydrogen and X is CR6R6' wherein R6 and R6' are
hydrogen
atoms, and R7 is dimethylamino); 9-substituted 4-dedimethylamino tetracycline
compound, wherein X is CR6R6', R4, R5, R6', R6, and R7 are hydrogen; and 9-
substituted
sancycline (wherein Ra is NR4'R4õ, Ra' and R4" are methyl; R5 and R7 are
hydrogen and
X is CR6R6' wherein R6 and e are hydrogen atoms).
In another embodiment, the invention pertains to tetracycline compounds
of formula 11-
R7 RS R4
00
R v I R NR2Rr
gage
OR10 0 OR" 0 0
wherein:
-11-
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X is CHC(R13Y'Y), CR6'R6, S, NR6, or 0;
R2, R4', R4", R'' and RT' are each hydrogen, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic,
heteroaromatic or a prodrug moiety;
RR is NR4'R4", alkyl, alkenyl, alkynyl, aryl, hydroxyl, halogen, or hydrogen;
R2', R3, R10, Rl l and R12 are each hydrogen or a pro-drug moiety,
R5 is hydroxyl, hydrogen, thiol, alkanoyl, aroyl, alkaroyl, aryl,
heteroaromatic,
alkyl, alkenyl, alkenyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,
alkylamino,
arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy;
R6 and R6' are independently hydrogen, methylene, absent, hydroxyl, halogen,
thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl,
alkylsulfonyl,
alkylamino, or an arylalkyl;
R7 is NR7'Rr, alkyl, alkenyl, alkynyl, aryl, hydroxyl, halogen, or hydrogen;
R$ is hydrogen, hydroxyl, halogen, thiol, alkyl, alkenyl, alkenyl, aryl,
alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;
R9 is -CH2NR9aR9b, or linked with R10 to form a furanyl ring;
R9a is hydrogen, alkyl, alkenyl,. alkynyl, alkoxy, alkylthio, alkylsulfinyl,
alkylsulfonyl, alkylamino, arylalkyl, aryl, heterocyclic,or heteroaromatic;
R9b is alkoxycarbonyl, arylaminocarbonyl, or aryloxycarbonyl;
R$ is hydrogen, hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl,
alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, or an arylalkyl;
R13 is hydrogen, hydroxy, alkyl, alkenyl, alkenyl, alkoxy, alkylthio,
alkylsulfinyl,
alkylsulfonyl, alkylamino, or an arylalkyl;
Y' and Y are each independently hydrogen, halogen, hydroxyl, cyan, sulfhydryl,
amino, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl,
alkylsulfonyl,
alkylamino, or an arylalkyl, and pharmaceutically acceptable salts, esters and
prodrugs
thereof.
In a further embodiment, R4 is NR4'R4"; X is CR6R6'; R7 is NR7'Rr, R2, R2',
R5,
R6, R6" Rs, R9, Rlo, R", and R12 are each hydrogen; and, e, R4", RT, and R7"
are each
lower alkyl. In another embodiment, R9a is alkyl, alkenyl, or arylalkyl.
Examples of R9b
include alkoxycarbonyl, alkaminocarbonyl, aryloxycarbonyl, and
arylaminocarbonyl. In
another embodiment,R9a and R9b are linked to form a heterocyle, e.g., a
substituted or
unsubstituted piperdinyl ring. In a further embodiment, the piperdinyl is
substituted
with one or more fluorines or halogenated alkyl groups, e.g., at the 2, 3, 4,
or 5 position.
In another embodiment, the R9 moiety is (4'trifluoromethyl-piperdin-l-yl)
methyl, (4',
4'-difluoro-piperdin-l-yl) methyl, or (4'-fluoropiperdin-l-yl) methyl.
In another embodiment, R4 is NR4'R4", R4' and R4 are methyl, R5 is hydroxyl
and X is CR6R6', wherein R6 is methyl and R6' is hydrogen, and R7 is hydrogen
In
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another embodiment, R9a is alkyl, alkenyl, or arylalkyl. In a further
embodiment, the
piperdinyl is substituted with one or more fluorines or halogenated alkyl
groups, e.g., at
the 2, 3, 4, or 5 position. In another embodiment, the R9 moiety is
(4'trifluoromethyl-
piperdin-1-yl) methyl, (4', 4'-difluoro piperdin-l-yl) methyl, or (4'-
fluoropiperdin-l-yl)
methyl.
In another further embodiment, R9a is substituted alkyl. Examples include
alkoxy substituted alkyl (e.g., -(CH2)2-O-CH3), alkenyl substituted alkyl
(e.g., -CH2-
CH=C(CH3)2, -CH2-C(CH3)=CHCH3, -CH2-CH=CH-phenyl, etc.), heterocyclic
substituted alkyl (e.g., -CH2-furanyl, -CH2-CH=CH furanyl, -CH2-pyridinyl,
optionally
substituted), cyano substituted alkyl (e.g., (CH2)2-CN, etc.), alkynyl
substituted alkyl
(e.g., -(CH2)2-C sCH, etc.), halogen substituted alkyl (e.g., (CH2)2-CF3,
(CH2)3-CF3, -
CH2-CF3, -CH2-CH2F, etc.), amido substituted alkyl (e.g., -CH2-C(=O)-N(CH3)2, -
CH2-
C(=O)-NH2, etc.), carbonyl substituted alkyl (e.g., CH2-C(=O)-CH3, -CH2-C(=O)-
C(CH3)3, etc.), hydroxy substituted alkyl (e.g., (CH2-CH(OH)-CH3, -CH2-
C(OH)(CH3)2,
etc.), -CH2-C(=N-O-CH3)-CH3, cycloalkyl (e.g., adamantyl;etc.).
In another embodiment, R9 is substituted or unsubstituted benzyl. Ina further
embodiment, R9a is substituted with one or more fluorines (e.g., at the 2, 3,
4, 5, or 6
positions).
In a further embodiment, R9b is hydrogen, substituted or unsubstituted alkyl
(e.g.,
methyl, ethyl, -CH2-CH=CH-furanyl, -CH2-CH=C(CH3)2, -(CH2)3-CF3, -(CH2)2-CH2F,
-
CH2-CH2F, -(CH2)2-CF3, -CH2-CF3, etc.).
In another further embodiment, R9a and 0 may be linked to form a pyrrolidinyl,
.piperazinyl, piperidinyl, pyrazinyl, azapanyl, thiomorpholinyl, morpholinyl,
ttrahydroquinolinyl, or a decahydroquinolinyl ring. The ring maybe substituted
with
one or more fluorines at the 2, 3, 4, or 5 position. The ring may also be
substituted with
one or more fluorinated alkyl groups (e.g., CH2F, -CHF2, CF3, etc.), cyano
groups,
hydroxy groups, alkyl groups (e.g., methyl, ethyl, spiro-cyclohexyl, t-butyl,
etc.),
heterocyclic (e.g., optionally substituted morpholinyl), thiol groups, alkoxy
groups,
alkyloxycarbonyl groups, carbonyl groups (optionally bonded directly to an
atom in the
ring), and exocyclic and endocyclic double bonds. In one embodiment, the ring
is
substituted with a =CF2 group. The ring may also be linked to a -O-(CH2)2-0-
group
which maybe attached to the pyrollidinyl or piperidinyl ring through one
carbons or
through two adjacent carbons.
When R9 is linked to R10 to form a furanyl ring, the ring can be further
subsituted, e.g., with phenyl or other substituents which allow the compound
of the
invention to perform its intended function.
In a further embodiment, the tetracycline compound is selected from the group
consisting of-
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H,C.N.CH, H,C.N.CFI, H3C.N.CH3 H,C.N.CH, HC-NCH, H'C.N.CHI
H H ~__ H_ H - H F{
NH,
OH O O O ON O OFD O P- ~,
cw, OH O O O
H .-CH, H3C.N.CH2 H,C.NCH3 H,C CHr,0.H cHI H,C N.CH,
CHN H ON H N ON
GGHa \ OH =
N I / \ NH, N/~N OH O OF1 O' ~C OH O oFP CH. CH, OH 00
H,C-N-CH, H O N-CH, ",%-CH. H,C-N.CH3 H,C.N.CH3 H,C.N.CH,
OH
N @ ON ~V-- HON WO&
H,C a Y ~F pig. N I / NHt OIZ OH O O OOH O HaC.tea Hac- N-c-+, -+,c-N- H,O-N- -
~c-N-
H H - OH F H _H OH H Li = OH
F1 F
~x N - NH6 F`- NHt
OH O OFP O ON O OFP 0
k OH 0
WP
H,C. CH, H,H .CH, H,C.N.CH3 H,c.N.CH, H,C..CH, H,C.CH,
jJ_ _ ON H H OH H H -
NHa
OH O HF0^t" ! H~ O C 0 ON O - x
H,C, .CH, H,C..CH, H,C.N-CH1 H,C.CH, H3C-N.CH3 H,C.N
Ha .CH,
VIC L-t1 U R H, H,C. CH, OH O CH6 _ OH 0 H Oti__,~'.AOb 01
ON O
H,C.N.CH3 H H,C.N-CW, H,C.N.CH, H,C.N.CH, H3C-N.CH3 H3C.N-CH,
H
ON
OH
H H - OH 6~1
0 NH, H2c NH, I / \ - I NH2
OH O O OH ON OH O O 0
H,C.NCHI H3C.N-CFI6 FyC.N.crt, H,C.N.cH, H,C=N.CH, H3C.N-CH,
H N OH H_ H OH ~x5 NC.ll- ~, OH O F~ 1IOII F O
H O O
N,C.N'Cfii
H,GN.CFt, k,c=N-CK H,C'N-CH, v N -K
F H kl 3 ON H N ON H H- ON
OH O H OFf OI F ON O O
, 7 ~0, Plr-
H3 CH, H %..CHI HaC HaC-N CH+ HiG CH, FIaC-N-Cf~a
H H e N 014 F H H_ -
~1 NH, - NFi, I/ a - NH1
at off O O O ON 0 ON O O.
H3C.N.CH3 H,A,N.CHi }~3C WON' H,C.N.CH, H,C.N.CH, 0...
_ - tl H O
9~ U_ U_ F` T N N
CDJ~
N NH, r'Y~ve
ON O F ON O'O O ON O
H,O.N-CHa FIaC^N-C~ F H+C, CK, HaC,N-CHa H,CrN-CHa Har^~I-CHa
H_ - F \ H_ H OH H H - OH
F
cxix A O ON OH O ON OF O OFf'O O
F
H,C.N.CH3 H,C.N.CH, F F H,C.N-CH3 H3C.N.CH3OH H,C,N.CH3 H1C.N.CH,
H e F H H e H H- OH
6NN N ~~ F ON O O
OH O OH O ON O Olp O
H3C.N-CH6 HIC=N-CH3 H,C.N.CH, H,C.N-CH, H,C,N.CH, H,C.N.CH,
//--O W=H~ ~~-- H OH /_ b,~ H H OH
0-0 1
OH O O OHOH O Oho 0 OH O OHO O
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WO 2005/009943 PCT/US2004/020249
-- H' HaG -CHa HaC- CH6 HaC- .CHa HaC. -' H,C-H-CHa
y ~Jl H H_ OH F NHs NH.
O OH O OH O OH 0'
N'C~ FIaC.N.CIia H FIaH =
F N_ - ~~ N
Ql-k ON
C-A OH ' - - I~
F~N I I NHz Fy - NH= FF~I/~NN - N"2
O
OH O O OH 00 O F OH 0
FIaC. "~Yl^,j~ HaC. +' bOHF Ha~N-tea Ha.N- 'NHc
0 CH3 ~t~. OH 0 O i H 01
HSC.N.CH8 H6C.N.CH3 Hac.N.CHl H HaCC.N.CHa H HaC.N.CH8
H H = - = ~~~NHCIH
a
Ha OH O O 0 OH 0 O F OH 0 0
C
FIaC.N.CH3 HaC N.CH3 H3C.N.CH3 H3C N.CHa NcC I.CF6 H HaH N.CH3
V H OH H ti - OH O OH
Qy
2N H3 Hc 916
OH 0 0 O' " 0 O F OH O O O OH 0 OFP O
F
%c N-CH. H3O-pr l+ HaC.N.CH. %C-N-Cla H3C. CH% Hac cH3
tl H v F H H- F N OH
co _ NHa o c tai. N c I NHZ
OH O OFP O OH O p OH O O O
HaC. CHa HaC .CHa Ha CHa NC CFIa HaC. CHa HaC .CHa
U u N N Ham- OH N " OH
w rya
O_ NHz Fp NHa Ha _ N
OH O OF 0 F OH 0 OFf 0 CHa OH O
HaC.N-CHa H3C.N.CHa H H3C,N,CH3 H3C.N.CHa H,C.M.cAO
OH H OH F F
NHs H ~~1,=NNz H O OO OH O OfQu0 H F 1` HaC.N.CHa H H3C.N-Chia HaC, CHa HaH
ICON
r r OH OH
NH2 011N
N i`,~~^oD
I ' O N w o' NH.
OH O O OH O O L~~.1 O OH O
HaC.N.CH3 HaC.N.CH8 /~ HaC.N.CH3 HVC.N.CH3 ~Ha HaC N_CHa
H = OH ) H H_ OH^ k! H
CN - NHZ \{ N p/~fp+c
~~QcH2 O Ot~ 0
H3G N.CH3 H,~. .CHa H,C. CH, H, rCt% c H8C
OH H~ - F Fla - b N OH
-- NH2 - NHa I _ _ Nkia
OH O ~i o OH O O O
F HaC.N.CHa HaCN.CH% H3C'WCH3 HaC N.CH3
H ~a H~ry Ca
off OH - off
F 1 `~1 N ~~ - NHz NHa
OH O OFP 0 NH H 0 OFP
V " OH O 0
HaC. ,CH3 F1aC. CH3 HaC. CHa N- .CHa Ha CHa H3C. .CH,
N H OH N OH SH N' FI Fi N OH
N _ `'NHa I NHzN NHi
OH O OFP 0 OH 0 0& O HaC pH 0 OI r 0
HaC.N.CHa ' N.CH' 1 0 N.CHa N.CHa
YSH H H pH N H OH = OH Cth
NHz
C H I - ' NHS HzN~/~ I I NHa ON~ _'
Y o
CHa OH O OH O O\1~~~
HP ~'r
H3C OH 0 OH O
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WO 2005/009943 PCT/US2004/020249
H6C-N.CHa H,C.~.CH, H,C-N.CH3 H3C.N.CH3 H,C-N,CH3 H3C,N.CH,
H H- OH OH O H H' OH
F
HHI ~C NH, -ON \ \ ~~NHz
OH O OH O OH O O O pH p pF{" b p
C
L~ 4 tl\N-C~ F -~ N ~N OH F HaG H QH\N.CHI
OH ON
NH2
1 OH O 01? O ON O O
P-O
HIC- - -CNa H,c CH3 H,c CH, F F ",c= H,L-N-
F H H OH F N N= off F !1 ~ = ON
N MH, HO I NH,
OH O O OH O Of? O OH O Oli O
H3C.N.CH, tw-U-C"3 No, H,C~ -
F r Cris N _ F r 4 s -C"3 CH.
OH _I OH
h'Yv_~'=,=1 I p_' I NH,
NHI
OHO Otf O ~ O O
H,C-N.CH, H, C- rc%
N N - OHt OH
_ = NH,N NH, H3C \ I \ NH2
OH , O OH O
OH O O
y N. \ CF~ 'N"di H,C.N CH, _ H, - H. H
ON ON _ OH
0H Q ~ ` NH
V=0 H H -
~
i^ r r~ - I NHt H,C- O O OH O
1s HyC a
0H O OH O O
1 `~
F p~~-+[r~'
V- OHS
_ OH OH
NH: Ha' 0 0
O OH O 0 O O O
H,N
N-OI HaC ~-CH,
~W \ -I 0
OH OH NH,
and pharmaceutically acceptable salts, est ers, and prodrugs thereof.
3. 7-Substituted Tetracycline Compounds
In one embodiment, the invention pertains to novel 7-substituted tetracycline
compounds.
The term "7-substituted tetracycline compounds" includes tetracycline
compounds with substitution at the 7 position. In one embodiment, the
substitution at
the 7- position enhances the ability of the tetracycline compound to perform
its intended
function, e.g., treat tetracycline responsive states. In an embodiment, the 7-
substituted
tetracycline compound is 7-substituted tetracycline (e.g., wherein R4 is
NR4'R4', R4' and
e are methyl, R5 is hydrogen and X is CR6R6', wherein R6 is methyl and R6' is
hydroxy); 7-substituted doxycycline (e.g., wherein R4 is NR4'R4 , R4' and R4"
are methyl,
R5 is hydroxyl and X is CR6R6', wherein R6 is methyl and R6' is hydrogen); 7-
substituted
tetracycline compound, wherein X is CR6R6', R4, R5, e'. and R6 are hydrogen;
or 7-
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substituted sancycline (wherein R4 is NR4'R4", R4' and R4"are methyl; R5 is
hydrogen
and X is CR6R6. wherein R6 and R6' are hydrogen atoms).
The invention pertains, at least in part, to 7-substituted tetracycline
compound of
Formula III:
R7 RS R4
x OR3
1 1 R9
ORu
RIO Rtt 0
wherein:
X is CHC(R13Y'Y), CR6'R6, C=CR6'R6, S, NR6, or 0;
R2, R , R4', and R4" are each independently hydrogen, alkyl, alkenyl, alkynyl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic,
heteroaromatic or a prodrug moiety;
R4 is NR4'R4", alkyl, alkenyl, alkynyl, aryl, hydroxyl, halogen, or hydrogen;
R2', R3, R' , Ru and R12 are each hydrogen or a pro-drug moiety;
R5 is hydroxyl, hydrogen, thiol, alkanoyl, aroyl, alkaroyl, aryl,
heteroaromatic,
alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfnyl, alkylsulfonyl,
alkylamino,
arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy,
R6 and R6' are each independently hydrogen, methylene, absent, hydroxyl,
halogen, thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy, alkylthio, alkylsuf
nyl,
alkylsulfonyl, alkylamino, or an arylalkyl;
R7 is substituted or unsubstituted pyrazolyl, furanyl, thiophenyl, or
thiazolyl;
R8 is hydrogen, hydroxyl, halogen, thiol, alkyl, alkenyl, alkynyl, aryl,
alkoxy,
alkylthio, alkylsulfnyl, alkylsulfonyl, alkylamino, or an arylalkyl;
R9 is hydrogen;
R13 is hydrogen, hydroxy, alkyl, alkenyl, alkynyl, alkoxy, alkylthio,
alkylsulfnyl,
alkylsulfonyl, alkylamino, or an arylalkyl; and
Y' and Y are each independently hydrogen, halogen, hydroxyl, cyano,
suliihydryl,
amino, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsufnyl, alkylsulfonyl,
alkylamino, or an arylalkyl, and pharmaceutically acceptable salts thereof
In a further embodiment, R4 is NR4'R4"; X is CR6R6', R2, R2', R5, R6, R6', Rs,
R9,
R' , Rtt, and R12 are each hydrogen; and, R4', and R4 are each lower alkyl,
e.g., methyl.
In one embodiment, the tetracycline compound is a doxycycline compound and
R7 is substituted or unsubstituted aminomethyl (e.g., -CH2NR7aRn').
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In one embodiment, R7 is substituted (e.g., N-alkyl substituted) or
unsubstituted
pyrazolyl. In another embodiment, R7 is diethyl amino. In another, R7 is
substituted
amino methyl. In a further embodiment, the substituted aminomethyl is
substituted with
a pentyl group (e.g., -CH2-C(CH3)3), two methyl groups, or fluorinated alkyl
(e.g.,
fluorinated propyl, e.g., -CH2-CH2-CF3).
In another embodiment, R7 is substituted phenyl. In a further embodiment, R7
is
phenyl substituted at the 5 position (of the phenyl ring) with an alkyl
substituted amino
methyl group (e.g., (-CH2- N(CH3)2, -CH2-NH-CH(CH3)2, -CH2-N(CH3)-CH(CH3)2, -
CH2-N-piperdinyl), -CH2NH-CH3a -CH2-NH-cyclopropyl, CH2-NH-t butyl, -CH2-
N(CH3)-benzyl, -CH2-N(CH3)-CH2-CH=CH2, CHi NH-(CH2)2-CF3i CH2-NH-CH2-
C(=O)-NH2, or -CH2 NH-cyclohexyl, ). In a further embodiment, the piperdine
may be
substituted at its 4 position (e.g., with fluorine, methyl, etc.).
In another embodiment, when R7 is a phenyl substituted at the 5 position with
an
alkyl substituted amino methyl group, the phenyl may also be substituted with
a fluorine
(e.g., at the 2, 3, 4, or 6 position) or an alkoxy (e.g., methoxy group) at
the 2, 3, 4, or 6
position.
In another embodiment, R7 is phenyl with a 2 -position amino alkyl
substituent.
In a further embodiment, the substituent is dialkylanunomethyl (e.g.,
dimethylaminomethyl, -CH2-N piperazinyl). In a further embodiment, the
piperazine is
substituted with one or more fluorine or methyl groups. In another further
embodiment,
the phenyl R7 is further substituted at the 3, 4, 5, or 6 position with a
methoxy group. In
another embodiment, the phenyl is linked to a methylene dioxy group through
its 4 and 5
positions.
In another embodiment, R7 is phenyl with a 4-position amino alkyl (e.g..,
aminomethyl) substituent. In a further embodiment, the aminoalkyl substituent
is -CH2-
NH-CH(CH3)2, -C(CH3)-NH-(CH2)2-CH2F, -CH2-NH-CH2-cyclohexenyl, -CH2-N-
piperidinyl, -CHZ N(CH3)-CH2-CH=CH2, or -CH2 NH-(CH2)2-CF3).
In another embodiment, R7 is phenyl substituted with a -C(=N-O-R)-R' group,
wherein R and R' are each alkyl. In a further embodiment, the substituent is
at the 4-
position of the phenyl ring. In another embodiment, R7 is phenyl substituted
at the 4-
position with an alkoxyalkyl group (-CH2-O-CH3). In another embodiment, R7 is
phenyl
substituted with an alkylcarbonylamino group.
In another embodiment, R7 is substituted furanyl. In a further embodiment, the
furanyl is attached at the 2-position of the furanyl ring. In a further
embodiment, the
furanyl is substituted with an amino alkyl, e.g., aminomethyl group at its 5-
position.
Examples of aminomethyl groups include: -CH2N(CH3)-CH2-C6H5, -CH2 N(CH3)-CH2-
CH=CH2, -CH2-N(CH3)-CH(CH3)2, or -CH2-N-piperidinyl. In another embodiment,
the
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WO 2005/009943 PCT/US2004/020249
furanyl is substituted at the 3-position, e.g., with an aminoalkyl
substituent. Examples
of such substituents include -CH2-N(CH3)2, -CHZ N piperidinyl
In another embodiment, R7 is substituted furanyl attached at its 3-position.
In a
further embodiment, the furanyl is substituted with an aminoalkyl substituent.
In
another further embodiment, the aminoalkyl substituent is -CH2 N piperazinyl
or -CH2-
N-(CH3)2-
In another embodiment, R7 is substituted or unsubstituted thiophenyl. In a
further embodiment R7 is is substituted with an aminoalkyl moiety. In another
further
embodiment, the aminoalkyl moiety is -CH2 N (CH3)2.
In another further embodiment, R7 is substituted pyridinyl. In a further
embodiment, R7 is attached to the phenyl ring at its 3 -position. In another
further
embodiment, it is substituted with a aminoalkyl moiety at its 5-position.
Examples of
aminoalkyl moieties include -CH2-N-(CH3)2, -CH2-N-piperidinyl, -CH2-N(CH3)-CH2
CH=CH2, Or -CH2-N(CH3)-CH(CH3)2.
In another further embodiment, R7 is alkylcarbonylaminoalkyl. In another
further embodiment, R7 is -CH2 NH-C(am)-CH3.
In another further embodiment, R7 is amino substituted alkenyl. In another
further embodiment, R7 is -CH=CH-CH2 N(CH3)2 or -CH=CH-M2N piperid nyL In
another embodiment, R7 is amino substituted alkynyl (e.g., -C M-CH2-N(CH3)-
(CH2)2-
CF3 or -C r=C-(CH2)rN-piperidinyI.
In another further embodiment, R7 is substituted -CH2-N-piperidinyl. In
certain
embodiments, the piperidinyl is substituted with one or more fluorines, e.g.,
at the 4-
position of the piperdine ring.
In another embodiment, the R7 substitutuent is alkylaminocarbonyL In a further
embodiment, the substituent is -C(0)-NH-(CH2)2-N(CH3)2.
In another further embodiment, the R7 substituent is aminoalkylcarbonyl. In a
further embodiment, the substituent is -C(=O)-CH2 N(CH3)2, -C(am)-CH2-NH-
(CH2)r
OCH3, -C(0)-CH2 N piperidinyl and -C(am)-CH2-N pyrollidinyl.
In another further embodiment, the R7 substituent is N-piperdinyl substituted
alkyl. In a further embodiment, the R7 substituent is -(CH2)4 N piperdinyl or -
(CH2)2-
N-piperdinyl.
In another embodiment, the R7 substituted is -(CH2)2-N(CH3)2 or C(0)-CH3.
In another further embodiment, the R7 substituent is aminoalkyloxycarbonyl.
Examples of aminoalkyloxycarbonyl substituents include C(=O)-O-(CH2)2 N-
piperdinyl
and -C(0)-O-(CH2)Z N(CH3)2.
In a further embodiment, the compounds of the invention are:
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-N HA
H3C.N,CHs Fi
C C H,(- CFt~
H OH sH.H,Ctis H,(- H N OH
I I ON
I- I NH2
I_ + NHZ Nf~ OH O OH 0 OH 0 0
0 OH 0 OHO 0
H+O/~ HZN C Ha \ H'C.N,CH'
NN off N O_y CH'
y H OH I
OH o O I/ \ - I NHZ H H OH
ON 0 OHO 0 I \p I NHZ
H20.p OH O OH 0
H CH, 9N HsC O H+O-N-CHs IWl CH. I \ H,C.wat
rHsNN- H H. I\ H_ H OH
p_ I Nit
NH2 O
0 OH O OH O
HH~Ctis
FtsC_ CFIs
H H N' OH Hs0. ,CH2
H H OH H Li N'oH
OHO O I _ NF6 NH2
yF~ OH O O 0 0110 OH 0
H. NF IWYCH' H2C7.~OH2
I ~ _ TT _
c
H H HsC y CHs
- - OH \ H HH H H_
OH O O O
OH O 0 ON O OFP
0 HA -CH4
O KC,,-CHs Ha Q% U G Hy OFI H PY OH
H2 OH 0
O
off 0 0 OH O O 0 QHs Hr- CHs
H,C-N O Hs0.trca
N U- H
NH2
0 oFP o t+ U tr:"N H,cH2
~ ~
p NH=
OH O off O OHO 0
n H
N N6jN%%rCH3 N.C ti~C,N.cH~
yCHs OH
H H \ -- I NHS
M OH O OI f O
0 0 0 -20-
CA 02744317 2011-06-22
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E CH3
N.CH3 ' IMF o
HC_ .CH3 N
N
44ou H,C .cH, I H3C,N_CH3
H H H H OH
NH,
I/ pp_ I NH,
OH 0 0I-r O NH3
OHO ,0 OH O ~OFi O
0 ' \ /l
cl,
HN" HNJ~ 6av-
H,C_ H3C, CH3 H3C_O t%C'N CH3 O H H !~" OH H=
NH=
0 11
- I NH3 F4 OH O 0 0 OH O 0
O
HN CH N,Ct%
OH N
H H
H k3H=N'CH3 / - I NH, OH
OH OH O O NH,
I / I NH' OH O OH O
OH O 0
CH3 H3C' NN NH= F
O
N HC,N,CH3 H,C H CH,
\ H H = OH \ H_ H
H Nc PYct a / NH3 q- NHz
OH off 0 011 o
p OH 0 OH 0
OH O 0 O F HUN'CH3
CHI
N,/,, CH2 N H3C,H=cf
- OH O NH H3C,N,CFi3
H_ H NH3
_
~ H H , / H3C,N,CH3 _ ~~I~NH.
H- OH OH 0
' OH 0 OH 0 O p O
Hz
OH O 0 P3C=NCH2
F
N H3C,N,CH3 i H, cH, \
H Hz OH H off H3C, CH3
H H_ OH
/ _. NH2 - NH3
NHz
OH 0 O OH O O I/ p
nOH O OFi 0
H3C,Ni,,,.pCH2 H3C,Ni"-'CH2
N / H C. CH H3C,NCH3 HF.Tc*%
H 3H N' 3 H H= H OH
\ - OH OH NH=
/ \p NH2 NH2 OH 0 IHI OH O OH O OH O OH O
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/NCH= n HaGN'~
O C H,C.N-CH, N O O H HsC.N.CHs
H O 0 H3 C_ -CH, OH
OH Ft H N H \p NH2
NH= O
\ _ I NH OH O OIi O
OH O O O a
OH O O
\ Ni-CH= H,C.N-CH3
~N'I O I a0N-cHa 0 f%CCN, -CH.
H,O rfq4, H=
OH H OH
OH \
l i I NH2 I \ I NH2
NHi -
off O O OH O OH O O
H,C,N.CH3 -CHs
H3 CH3 Ntia4
H_ H = CI1, Cli,
4'0~v ~di O NH, H OH
OH O OH" O ~
o OH O O
/C16si,QN.CH, HN N~CHa
N~\'CH2 6OMP Fs
H H = OH \
NH, \ H H N OH
OH 0 O
NH, I \ NH.
OH 0&. O
06
~o
N F F
O H,C.~-CHs HN C`NICH3
H H - OH O ' .N-CH3 H = OH
H H - OH
Nl-~ NH,
OH O 0 \ NH, OH O O
OH O O
/~ H3C-N-CH3
H'C.N'CN H3C. -CH, H H OH
N'
- NH=
4 \
T OH
H- H H N~OOCNH
off o ori' NH2 OH O Oti O
OH O OHP O
,aia
CHs
HsC. CHs H,a H, c, -CH. H H - OH
H H OH N tl V off I \
NHS '
H
0 o OH O O
OH 0 0 0
O CH3 H,0. -CHs H,C~
\ H H N OH H,C. cls
0 HsC_ CHs
NH, HsC, Ctls H H N OH
OH 0 OHO O H H H
NH,
OH O OttQ o o OH O 0 O
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YN, CH, F G
H HaH'
HaC-N^ H,4J,
F _ - off
H H_ - HH. p
OHO OH O NH:
VYC.
H.
GO
Ha H C,_j= OH H. KC-0 ~~~=
{ _- ON
C H
N
O Ha CHs rHWOFHPOP
O O OHO
o HHa
O
H.C.N_Ct%
6(H.CINWICH3 HN- CH, H C'O HN
Ha0.N.C %
H HOH NH2
NH2 oHo o / \Q NH,
OH 0 OH 0
HaG a HN X)
~' HN Hip- CH4HC '
HaC,WGla
H t CH.
93CN HCH2
H -
NHa OH O / _ NHt
NH, OHO O o
O
H N 6
N
O H3C.N'CHa N
H OH I / H,C~N.CH, 16-C
41
H H OH
OH
OH O OH O ~/ gyp. i NHZ OH O OH 0 0
V NHa
40.~
OH and pharmaceutically acceptable esters, prodrugs, and salts thereof.
4. 8-Substituted Tetracycline Compounds
The invention also pertains, at least in part to 8-substituted tetracycline
compounds.
The term "8-substituted tetracycline compounds" includes tetracycline
compounds with substitution at the 8- position. In one embodiment, the
substitution at
the 8- position enhances the ability of the tetracycline compound to perform
its intended
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function, e.g., treat tetracycline responsive states. In an embodiment, the 8-
substituted
tetracycline compound is 8-substituted tetracycline (e.g., wherein R4 is
NR4'R4"; R4' and
R4" are methyl, R5 is hydrogen and X is CR6R6', wherein R6 is methyl and R6'
is
hydroxy); 8-substituted doxycycline (e.g., wherein R4 is NR4'R4"; R4' and R4"
are methyl,
R is hydroxyl and X is CR 6', wherein R6 is methyl and R is hydrogen); or 8-
substituted sancycline (wherein R4 is NR4R4'; R4' and e are methyl; R5 is
hydrogen
and X is CR6R6' wherein R6 and R6' are hydrogen atoms. In an embodiment, the
substitution at the 7 position of the 8-substituted tetracycline compound is
not chlorine
or trimethylamino. In one embodiment, R4 is hydrogen.
In one embodiment, the 8-substituted tetracycline compound is of formula IV:
R1 RS R~
R / X W
\ I \ ( 2RF
R9 =
OW2
OR10 O R" O O lj = J
wherein:
X is CHC(R13Y'Y), CR6'R6, S, NR6, or 0;
R2, R4', R4", R7' and R7" are each hydrogen, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfnyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic,
heteroaromatic or a prodrug moiety;
R4 is NR4R4", alkyl, alkenyl, alkynyl, aryl, hydroxyl, halogen, or hydrogen,
R2', R3, Rio, R'1 and Rte are each hydrogen or a pro-drug moiety;
R5 is hydroxyl, hydrogen, thiol, alkanoyl, aroyl, alkaroyl, aryl,
heteroaromatic,
alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfnyl, alkylsulfonyl,
alkylamino,
arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy;
R6 and R6' are independently hydrogen, methylene, absent, hydroxyl, halogen,
thiol, alkyl, alkenyl, alkynyl, aryl, alkoxy, alkylthio, alkylsulfinyl,
alkylsulfonyl,
alkylamino, or an arylalkyl;
R7 is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl, alkynyl,
aryl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, arylalkyl, amino,
arylalkenyl, arylalkynyl,
acyl, aminoalkyl, heterocyclic, thionitroso, or -(CH2)a3 (NRlc)aiC(=W')WR7B;
R$ is substituted phenyl or substituted pyridinyl;
R9 is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl, alkynyl,
aryl,
alkoxy, alkylthio, alkylsuffmyl, alkylsulfonyl, arylalkyl, amino, arylalkenyl,
arylalkynyl,
acyl, aminoalkyl, heterocyclic, thionitroso, or-(CH2)a3NR9cC(=Z')ZR9a;
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R7a, e, R'0, R7d, R7e, R' , R9a, R9b, R9c, R9d, R9e, and Rsf are each
independently
absent, hydrogen, acyl, alkyl, alkenyl, alkynyl, alkoxy, alkyltbio,
alkylsulfnyl,
alkylsulfonyl, alkylamino, arylalkyl, aryl, heterocyclic, heteroaromatic or a
prodrug
moiety;
W is CR7dR7e, S, 0 or NRT;
W' is 0, NR~ or S;
R13 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, alkylsulfnyl,
alkylsulfonyl,
alkylamino, or an arylalkyl, and pharmaceutically acceptable salts, esters and
prodrugs
thereof.
In a further embodiment, the invention pertains to compounds wherein Xis
CR6R6'; R2, R2=, R6, R6', R8, R10, R", and R12 are each hydrogen; R4 is
NR4'R4"; R4. and
R4" are lower alkyl; and R5 is hydroxy or hydrogen-
In a further embodiment, R8 is substituted phenyl, e.g., o-substituted phenyl,
e.g.,
aminomethyl substituted phenyl. In a further embodiment, the 8-substituted
tetracycline
compound is:
OH
F~~N ~ ~ _ I HH=
OH 0 ' and pharmaceutically acceptable salts, esters, and prodrugs
thereof..
In another further embodiment, R8 is substituted pyridinyl, e.g., halo-
substituted
pyridinyl, e.g., 6-fluoro-pyrindin-3-yl. In a further embodiment, R9 is amino.
In yet a
further embodiment, the 8-substituted tetracycline compound is:
F N FI,C rCH,,
H- OH
FIiN
H o o and pharmaceutically acceptable salts, esters, and prodrugs
thereof..
5. 13-Substituted Methacycline Compounds
In one embodiment, a 13-substituted tetracycline compound is of formula V:
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R13
R7 R5 R4
Rs
OW
R9 =
ORIZ
OR10 0 OR" O O (V)
wherein:
R2, R4', R4", RT and RT' are each hydrogen, alkyl, alkenyl, alkynyl, alkoxy,
alkylthio, alkylsulfinyl, alkylsulfonyl, alkylamino, arylalkyl, aryl,
heterocyclic,
heteroaromatic or a prodrug moiety,
R4 is NR4'R4", alkyl, alkenyl, alkynyl, aryl, hydroxyl, halogen, or hydrogen;
R2 , R3, Rio, R" and R12 are each hydrogen or a pro-drug moiety,
R5 is hydroxyl, hydrogen, thiol, alkanoyl, aroyl, alkaroyl, aryl,
heteroaromatic,
alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl,
alkylamino,
arylalkyl, alkyl carbonyloxy, or aryl carbonyloxy,
R7 is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl, alkynyl,
aryl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, arylalkyl, amino,
arylalkenyl, arylalkynyl,
acyl, aminoalkyl, heterocyclic, thionitroso, or-(CH2)a3 (NR7c)o-iC(=W7)WR7a;
R8 is substituted phenyl or substituted pyridinyl;
R9 is hydrogen, hydroxyl, halogen, thiol, nitro, alkyl, alkenyl, alkynyl,
aryl,
alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, arylalkyl, amino,
arylalkenyl, arylalkynyl,
acyl, aminoalkyl, heterocyclic, thionitroso, or -(CH2)o.3NR9cC(=Z')ZR?a;
R7a, R7', R7c, R7d, R'e, R7, R9a, R91i, R9c, R9d, R9e, and R81 are each
independently
absent, hydrogen, acyl, alkyl, alkenyl, alkynyl,.alkoxy, alkylthio,
alkylsulfinyl,
alkylsulfonyl, alkylamino, arylalkyl, aryl, heterocyclic, heteroaromatic or a
prodrug
moiety;
W is CR7dR7c, S. O or NO;
W' is 0, NR7 or S;
R13 is 4-alkyl substituted phenyl, and pharmaceutically acceptable salts,
esters
and prodrugs thereof.
In a further embodiment, the invention pertains to compounds wherein R2, R2',
R8, R1 , Rl1, and R12 are each hydrogen; R4 is NR4'R4"; R4' and kr are lower
alkyl; and
R5 is hydroxy or hydrogen.
In a further embodiment, the phenyl R13 group is substituted with an
aminomethyl substituent. In another further embodiment, the aminomethyl
substituent
is dimethylaminomethyl. In another further embodiment, the invention pertains
to
compounds of the formula:
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H3C.N
CHa I,C.N.CH3
H _ OH
(, _ I NHZ
OH O O O
and pharmaceutically acceptable salts, esters, and prodrugs thereof_
In one embodiment, the tetracycline compounds of the invention do not include
those described in U.S.S.N. 09/660,598, 09/823,884, 09/852,908, 10/819,343,
10/820,456, 09/894,805, 09/895,796, 09/895,812, 09/895,797, 09/895,857,
10/097,634,
10/759,484, 10/337,914,10/636,437, 10/752,378, or 10/740,961.
6. Methods for Synthesizing Tetracycline Compounds of the Invention
The tetracycline compounds of this invention can be synthesized using the
methods described in the Schemes and/or by other techniques known to those of
ordinary skill in the art.
The substituted tetracycline compounds of the invention can be synthesized
using the methods described in the following schemes and by using art
recognized
techniques. All novel substituted tetracycline compounds described herein are
included
in the invention as compounds.
x m. + wlj~
Ic
u
Pt H2 P& III
oM
IF Yc
00
~H u
SCHEME1
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9- and 7- substituted tetracyclines can be synthesized by the method shown in
Scheme 1. As shown in Scheme 1, 9- and 7-substituted tetracycline compounds
can be
synthesized by treating a tetracycline compound (e.g., doxycycline, 1A), with
sulfuric
acid and sodium nitrate. The resulting product is a mixture of the 7 -nitro
and 9-nitro
isomers (1B and 1C, respectively). The 7-nitro (1B) and 9- nitro (1C)
derivatives are
treated by hydrogenation using hydrogen gas and a platinum catalyst to yield
amines 1D
and 1E. The isomers are separated at this time by conventional methods. To
synthesize
7- or 9-substituted alkenyl derivatives, the 7- or 9-amino tetracycline
compound (lE and
1F, respectively) is treated with HONO, to yield the diazonium salt (1G and
1H). The
salt (1 G and 1H) is treated with an appropriate reactive reagent to yield the
desired
compound(e.g., in Scheme 1, 7-cyclopent-l-enyl doxycycline (1H) and 9-
cyclopent-l-
enyl doxycycline (11)).
. "g NM.h
Ha
XWO,
oN O ox O 0
28
R Ny7R
I'mo' gyp{
Oak
m
O 0
WO
x
"Pak
wit
0
SCHEME 2
As shown in Scheme 2, tetracycline compounds of the invention wherein R7 is a
carbamate or a urea derivative can be synthesized using the following
protocol.
Sancycline (2A) is treated with NaNO2 under acidic conditions forming 7-nitro
sancycline (2B) in a mixture of positional isomers. 7-nitrosancycline (2B) is
then
treated with H2 gas and a platinum catalyst to form the 7-amino sancycline
derivative
(2C). To form the urea derivative (2E), isocyanate (2D) is reacted with the 7-
amino
sancycline derivative (2C). To form the carbamate (2G), the appropriate acid
chloride
ester (2F) is reacted with 2C.
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'CH3
FmodiN H
H 3B H
N,42 Fmoc-NCS NHZ
HOH H HO
3A 3C
~b
jj2 H i 3E s'
1) -Fmoc
R' H
HZ
H
H OH HO
3F
3D
SCHEME 3
As shown in Scheme 3, tetracycline compounds of the invention, wherein R7 is a
heterocyclic (i.e. thiazole) substituted amino group can be synthesized using
the above
protocol. 7-amino sancycline (3A) is reacted with Fmoc-isotbiocyanate (3B) to
produce
the protected thiourea (3C). The protected thiourea (3C) is then deprotected
yielding the
active sancycline thiourea (3D) compound. The sancycline thiourea (3D) is
reacted with
an a-haloketone (3E) to produce a thiazole substituted 7-amino sancycline
(3F).
cH,h h
(CH3)2
H NI~H/FfCI OH
HI or H H
x Pd/C
H H O O H H H OH
4A 4B IC
SCHEME 4
7- alkenyl tetracycline compounds, such as 7-alkynyl sancycline (4A) and 7-
alkenyl sancycline (4B), can be hydrogenated to form 7-alkyl substituted
tetracycline
compounds (e.g., 7-alkyl sancycline, 4C). Scheme 4 depicts the selective
hydrogenation
of the 7- position double or triple bond, in saturated methanol and
hydrochloric acid
solution with a palladium/carbon catalyst under pressure, to yield the
product.
ti
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NMe2
OH
N-iodosuccinimide OH
NH2 NH2
OH O OHIO O OH O OHIO O
5A 5B
x
Pd(OAc}2, Na2COy McOH I
NMe2
= OH
~ OH
B\OH NH2
SC OH O OH O O
5D
SCHEME 5
In Scheme 5., a general synthetic scheme for synthesizing 7 -position aryl
derivatives is shown. A Suzuki coupling of an aryl boronic acid with an
iodosancycline
compound is shown. An iodo sancycline compound (5B) can be synthesized from
sancycline by treating sancycliine (5A) with at least one equivalent N-
iodosuccinimide
(NIS) under acidic conditions. The reaction is quenched, and the resulting 7-
iodo
sancycline (5B) can then be purified using standard techniques known in the
art. To
form the aryl derivative, 7-iodo sancycline (5B) is treated with an aqueous
base (e.g.,
Na2CO3) and an appropriate boronic acid (5C) and under an inert atmosphere.
The
reaction is catalyzed with a palladium catalyst (e.g., Pd(OAc)2). The product
(5D) can
be purified by methods known in the art (such as HPLC). Other 7-aryl, alkenyl,
and
alkynyl tetracycline compounds can be synthesized using similar protocols.
The 7-substituted tetracycline compounds of the invention can also be
synthesized using Stille cross couplings. Stille cross couplings can be
performed using
an appropriate tin reagent (e.g., R-SnBu3) and a halogenated tetracycline
compound,
(e.g., 7-iodosancycline). The tin reagent and the iodosancycline compound can
be
treated with a palladium catalyst (e.g., Pd(PPh3)2C12 or Pd(AsPh3)2C12) and,
optionally,
with an additional copper salt, e.g., CaL The resulting compound can then be
purified
using techniques known in the art.
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R N(CH )2
6B H
TraaslUOU Mctd 7XIPZJNE
2~z H O
(
OH Phm b6vLpnd 6C
ONH2
H H HO TrnzIoiMwi
cdolyzt R
6A PbOVMWLtW
(CHA
H
6D
H NH2
6H
SCHEME 6
The compounds of the invention can also be synthesized using Heck-type
cross coupling reactions. As shown in Scheme 6, Heck type cross-couplings can
be
performed by suspending a halogenated tetracycline compound (e.g., 7-
iodosancycline,
6A) and an appropriate palladium or other transition metal catalyst (e.g.,
Pd(OAc)2 and
Cul) in an appropriate solvent (e.g., degassed acetonitrile). The substrate, a
reactive
alkene (6B) or alkyne (6D), and triethylamine are then added and the mixture
is heated
for several hours, before being cooled to room temperature. The resulting 7-
substituted
alkenyl (6C) or 7-substituted alkynyl (6E) tetracycline compound can then be
purified
using techniques known in the art.
6UV(CH3)2 CI U(CH3h
H MeowIia OH
H Y NH2 NEI2
H OH
7A 7B
SCHEME 7
To prepare 7-(2'-Chloro-alkenyl)-tetracycline compounds, the appropriate 7-
(alkynyl)-sancycline (7A) is dissolved in saturated methanol and hydrochloric
acid and
stirred. The solvent is then removed to yield the product (7B).
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0
F~,
CK~No Cw RI H3C' ,CH3
H carboxylic acid OH
NH: ""6
anhydrous
R bH
H B OH HO O
O OH O O
8A 813
SCHEME 8
As depicted in Scheme 8, 5-esters of 9- substituted tetracycline compounds can
be formed by dissolving the 9- substituted compounds (8A) in strong acid (e.g.
HF,
methanesulphonic acid, and trifluoromethanesulfonic acid) and adding the
appropriate
carboxylic acid to yield the corresponding esters (8B).
As shown in Scheme9 below, 7 and 9 aminomethyl tetracyclines may be
synthesized using reagents such as hydroxymethyl-carbamic acid benzyl ester.
H2N i (CH3)2
H
(CH3)2 ~Vd H -H Ip H2
H TFA H li
H2 +
~I(CH3)2
OH 24hr.,25C H
H2 NH2
H If
SCHEME 9
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 frther 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., Cl-C6 for straight chain, C3-C6 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 terra C1-C6 includes alkyl groups containing 1 to 6 carbon
atoms.
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
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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, suifamoyl, 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.
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, cyan, 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).
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.
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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., C2-C6 for straight chain, C3-C6 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 C2-C6
includes
alkenyl groups containing 2 to 6 carbon atoms.
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, sulthydryl,
alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfmyl, sulfonato, sulfamoyl,
sulfonamido,
nitro, trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromatic
or
heteroaromatic moiety.
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.
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., C2-C6 for straight chain, C3-C6
for
branched chain). The term C2-C6 includes alkynyl groups containing 2 to 6
carbon
atoms.
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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, cyan, amino (including alkyl amino, dialkylamino,
arylamino, diarylamino, and alkylarylamino), arylamino (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.
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.
The term "acyl" includes compounds and moieties which contain the acyl radical
(CH3CO-) or a carbonyl group. It includes substituted acyl moieties. 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, cyan, amino (including alkyl amino, dialkylamino,
arylamino, diarylamino, and alkylarylaxnino), acylamino (including
alkylcarbonylamino,
arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl,
alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,
sulfonamido,
nitro, trifluoromethyl, cyan, azido, heterocyclyl, alkylaryl, or an aromatic
or
heteroaromatic moiety.
The term "acylamino" includes moieties wherein an acyl moiety is bonded to an
amino group. For example, the tenn includes alkylcarbonylamino,
arylcarbonylamino,
carbamoyl and ureido groups.
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.
The terms "alkoxyalkyl", "alkylaminoalkyl" and "thioalkoxyalkyl" include alkyl
groups, as described above, which further include oxygen, nitrogen or sulfur
atoms
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replacing one or more carbons of the hydrocarbon backbone, e.g., oxygen,
nitrogen or
sulfur atoms.
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), aclamino (including
alkylcarbonylamino,
arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulthydryl,
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.
The term "amine" or "amino" includes compounds where a nitrogen atom is
covalently bonded to at least one carbon or heteroatom. The term includes
"alkyl
amino" which comprises 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 tern
"arylamino"
and "diarylamino" include groups wherein the nitrogen is bound to at least one
or two
aryl groups, respectively. The term "alkylarylan'iino," "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.
The term "amide," "amido" 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 and arylcarbonylamino
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."
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Amides also include urea groups (aminocarbonylamino) and carbamates
(oxycarbonylamino).
The term "carbonyl" or "carboxy" includes compounds and moieties which
contain a carbon connected with a double bond to an oxygen atom. The carbonyl
can be
further substituted with any moiety which allows the compounds of the
invention to
perform its intended function. For example, carbonyl moieties maybe
substituted with
alkyls, alkenyls, alkynyls, aryls, alkoxy, aminos, etc. Examples of moieties
which
contain a carbonyl include aldehydes, ketones, carboxylic acids, amides,
esters,
anhydrides, etc.
The term "thiocarbonyl" or "thiocarboxy" includes compounds and moieties
which contain a carbon connected with a double bond to a sulfur atom.
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.
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.
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 -0-.
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.
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,
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aryoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,
alkoxycarbonyl,
alkylaminoacarbonyl, arylalkylaminocarbonyl, alkenylaminocarbonyl,
alkylcarbonyl,
arylcarbonyl, arylalkyl carbonyl, alkenylcarbonyl, aminocarbonyl,
alkylthiocarbonyl,
alkoxyl, phosphate, phosphonato, phosphinato, cyano, amido, amino (including
alkyl
amino, dialkylamino, aryyamino, diarylamino, and alkylarylamino), acylamino
(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),
amidino,
imino, suithydryl, alkylthio, arylthio, thiocarboxylate, sulfates,
alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,
alkyl,
alkylaryl, or an aromatic or heteroaromatic moiety.
/ The term "heteroatom" includes atoms of any element other than carbon or
hydrogen. Preferred heteroatoms are nitrogen, oxygen, sulfur and phosphorus.
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. (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 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, 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.
It will be noted that the structure of some of the tetracycline 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.
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7. Methods for Treating Tetracycline Responsive States
The invention also pertains to methods for treating a tetracycline responsive
states in subjects, by administering to a subject an effective amount of a
tetracycline
compound of the invention (e.g., a compound of Formula I, II, III, IV, V or
otherwise
described herein), such that the tetracycline responsive state is treated.
The term "treating" includes curing as well as ameliorating at least one
symptom
of the state, disease or disorder, e.g., the tetracycline compound responsive
state.
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 tetracycline compound of the invention,
e.g., a 3,
10, and/or 12a substituted tetracycline compound. 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 characheterized by unwanted cellular
proliferation, including, but not limited to, those described in U.S.
6,100,248), arthritis,
osteoporosis, diabetes, and other states for which tetracycline compounds have
been
found to be active (see, for example, U.S. Patent Nos. 5,789,395; 5,834,450;
6,277,061
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)).
In a further
embodiment, the tetracycline responsive state is not a bacterial infection. In
another
embodiment, the tetracycline compounds of the invention are essentially non-
antibacterial. For example, non-antibacterial tetracycline compounds of the
invention
may have MIC values greater than about 4 pg/ml (as measured by assays known in
the
art and/or the assay given in Example 2).
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, cytoldnes, 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.
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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.
IPAF'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.
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.
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., a 3, 10, and/or 12a substituted tetracycline compound. The
disorders,
diseases and states described in U.S. Patents Nos. 6,231,894; 6,015,804;
5,919,774; and
5,789,395 are also included as NO associated states. -
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, osteoartbritis,
rheumatoid
arthritis, acute, recurrent and chronic infections (bacterial, viral and
fungal); 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 tendonitis);
uremic pericarditis;
acute and chronic cholecystis; cystic fibrosis, acute and chronic vaginitis;
acute and
chronic uveitis; drug reactions; insect bites; burns (thermal, chemical, and
electrical);
and sunburn.
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The term "inflammatory process associated state" also includes, in one
embodiment, matrix metalloproteinase associated states (MMPAS). NIMPAS include
states charachterized by abberrant 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., 3, 10, and/or 12a substituted tetracycline
compounds.
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, I
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
M WAS 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..
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.
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,
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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 maybe 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.
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,
amylotrophic
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).
Other examples of tetracycline compound responsive states are described in WO
03/005971A2, U.S.S.N. 60/421,248, and U.S.S.N. 60/480,482.
The language "in combination with" another therapeutic agent or treatment
includes co-administration of the tetracycline compound, (e.g., inhibitor) 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 an IPAS. 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
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invention include those described in U.S. Patent Nos. 6,100,248; 5,843,925;
5,837,696;
or 5,668,122,
In another embodiment, the tetracycline compound responsive state is diabetes,
e.g., juvenile diabetes, diabetes mellitus, diabetes type I, or diabetes type
H. 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. Patents Nos. 5,929,055; and 5,532,227,
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. Patents Nos. 5,459,135; 5,231,017;
5,998,390;
5,770,588; RE 34,656; 5,308,839; 4,925,833; 3,304,227; and 4,666,897.
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.
The invention also pertains to a method for treating acute lung injury by
administering a substituted tetracycline compound of the invention.
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
emphesema. In a
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further embodiment, the tetracycline compounds of the invention used to treat
acute
and/or chronic lung disorders such as those described in U.S. Patents No.
5,977,091;
-
6,043,231; 5,523,297; and 5,773,430!b'
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
tetracycline compounds of the invention are used to treat such disorders as
described in
U.S. Patents No. 6,231,894; 5,773,430; 5,919,775 or 5,789,395.
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, mucusae) 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 gellatinolytic 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 used to
treat a skin wound, and other such disorders as described in, for example,
U.S. Patent
Nos. 5,827,840; 4,704,383; 4,935,412; 5,258,371; 5,308,83915,459,135;
5,532,227; and
6,015,804a
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. Patent Nos. 6,043,225 and 5,834,449.
Bacterial infections maybe 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
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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, 2nd edition, Villanova, PA (1990).
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.
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.
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.
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.
8. Pharmaceutical Compositions of the Invention
The invention also pertains to pharmaceutical compositions comprising a
therapeutically effective amount of a tetracycline compound (e.g., a compound
of
Formula 1,11,111, IV, V or any other compound described herein) and,
optionally, a
pharmaceutically acceptable carrier.
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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
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.
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-hydro)y-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 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.
The preparation of other tetracycline compounds of the invention not
specifically
described in the foregoing experimental section can be accomplished using
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combinations of the reactions described above that will be apparent to those
skilled in
the art.
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 maybe 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 maybe formed with pharmaceutically
acceptable cations by conventional methods. Thus, these salts maybe 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.
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.
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.
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,
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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.
The tetracycline 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 (e.g., aerosols, etc.),
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.
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. The compositions of the invention may be formulated such
that
the tetracycline compositions are released over a period of time after
administration.
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
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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.
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.
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.
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.
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.
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'
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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.
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 pracctitioner 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.
Furthermore, the invention also pertains to the use of a tetracycline compound
of
formula I, II, III, N, V, or any other compound described herein, for the
preparation of a
medicament. 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.
EXEMPLIFICATION OF THE INVENTION
.25 Example 1: Synthesis of Selected Compounds of the Invention
F
N
H3C1 .CH3
H H N OH
H3
C~iN I / \ = ( NH2
OH O OH O
The above compound was prepared from 7-iodo-sancycline (15.0g, 22.9mmol)
combined with Pd(dppf)2C12 (1.7g, 2.29mmol) and DMF (300mL) in a 1L round
bottom
2 neck flask. Na2CO3 (7.2g, 68.2mmol) was dissolved in water (15mL) was added
to
reaction solution. 2-fluoro-pyridine-5-boronic acid (6.4g, 45.9mmol) was
dissolved in
DMF (25mL) and also added to reaction solution. Reaction mixture was stirred
at 65 C
(oil bath temperature) under an argon atmosphere and reaction was monitored by
HPLC
and LCIMS. Reaction shown to be complete within 3hr. Filtered through celite
and
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evaporated solvent in vacuo. Redissolved in MeOH (30mL) and precipitated in
MTBE
(3L) to produce a yellow precipitate. Filtered and dried under vacuum
overnight to yield
15g of yellow powder. This crude material (9g, 17.8mmol) was dissolved in
TFA/Triflic acid (83mIJ7mL) and cooled to 0 C using an ice bath. N-iodo-
succinimide
(8g, 35.6mmol) was added portionwise to reaction solution over 2hr. Reaction
complete
after 3hrs - and 20% more NIS added to reaction. Evaporated TFA in vacuo and
precipitated remaining acid in MTBE (1.4L) at room temp. Yellow precipitate.
Filtered
and dried under vacuum overnight to yield 8.4g of crude product. This crude
material
(4g, 6.3mmol) was combined with NaOAc (0.52g, 6.3mmol) in an oven-dried 25OmL
2
neck round bottom flask. Anhydrous DMF (6OmL) was syringed into reaction
flask.
Stirred under argon at room temp lhr. Diluted with more anhydrous DMF (120mL)
and
a CO-filled balloon was placed on top neck of reaction flask. CO was purged
through
reaction direction from lecture bottle for 15min. Flask then open to CO-filled
balloon
and allowed to stir at 60 C (oil bath temp) while Pd(PPh3)4 (2.2g, 1.9mmol)
was added
as a DMF slurry via syringe. Stirred at temperature Ihr. SnBu3H (1.6g,
6.3mmol) was
added via syringe pump over 2hr. Reaction monitored by HPLC and LC/MS and
shown
to be complete upon completion of tin addition. Evaporated solvent in vacuo.
Purified
by preparative HPLC in 20% yield in preparation for final synthesis step. This
purified
material (0.25g, 0.46mmol) was combined with anhydrous DMF (15mL) in an oven-
dried 100mL flask. InC13 (0.005g, 0.023mmol), N-methyl-allylamine (0.178,
0.23mmol)
were added to reaction and stirred at room temperature under argon lhr.
NaCNBH3
(0.035g, 0.55mmol) was added to reaction solution and was monitored by HPLC
and
LC/MS. Reaction 80% complete within 6hrs of reaction time. Evaporated solvent
in
vacuo. Final product was isolated by preparative HPLC in 10% yield as a yellow
solid.
ESI-MS: m/z (M + H) 593.
7-Ethyl-9-(4'. 4' Difluoro-N-Piperidinyl methyl)-Sang cy line
CH3 H3C..N.CH3
F H H -
F
N NH,
OH O OH O
The compound was prepared from 7-ethyl-9-formyl-sancycline (0.23g,
0.49mmol) combined with IUCl3 (0.011g, 0.049mmol), 4,4-difluoropiperidine.HCI
(0.17g, 0.98mmol), Et3N (0.099g, 0.98mmol), and DMF (8mL) in a glass vial.
Stirred
under argon at room temperature 30min. NaCNBH3 (0.043g, 0.69mmol) was added to
reaction vial and continued to stir at room temperature under argon. Reaction
was
monitored by LC/MS and HPLC and shown to be complete in 2hrs. Quenched
reaction
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with McOH (15mL) and evaporated solvent in vhcuo. Product was isolated by
preparative BPLC in 20% yield as a yellow solid. ESI-MS: m/z (M + H) 576.
7-(Trifluoroalkenyl)-9-(2'- trans-2-methyl-2-butene) aminomethy1 Sanccline
F
F F H3C,N.CH3
H H = OH
CH3
H3C~,N I _ ' NH2
OH O OHO 0
To a stirred solution of powered Zn (5.00 g, 76.5 mmol) in dry THE (50.0 mL)
at
0 C was added iodo-trifluor alkene (2.00 mL, 4.50 g, 21.0 mmol) slowly over a
0.5 h
time period. The reaction was stirred for an additional 1.5 h before it was
filtered under
an inert atmosphere and reduced of all solvent using rotary evaporation (25.0
C, 5.00
mm Hg) to yield the trifluoro-zinc-iodo-aikene reagent (approximately 3 mL).
Dry
DMF (10 mL) was added to the above zinc-reagent and this solution was added to
a
stirred solution of 7-Iodo-9-trans-2-methyl-2 butene sancycline free base
(1.00 g, 1.57
mmol) and tetralds(triphenylphosphine)palladium (0.181 g, 0.156 mmol) in dry
DMF
(10 mL). The contents were heated to 40 C and allowed to stir for 20 minutes.
The
reaction was then filtered and purified using reverse phase HPLC to give 7-
trifluoroalkene sancycline product (557 mg, 0.0942 mmol, 60% yield) LCMS m/z =
592.2392 (M + H).
7-(2' P E, zinyl)-9-(3',3',3'-Trifluoro-propylamino)-methyl-Sancycline
N iN HC. CH
H 3H N' 3
H OH
F~~1/~2~N NHZ
FF OH O OIPO O
Step 1:
7-Iodo-9-aminomethyl sancycline (569 mg, 1 mmol), indium trichloride (22 mg,
0.1 mmol) and trifluoropropionaldehyde (224 L, 2 mmol) were taken in DMF (25
mL)
and stirred at room temperature for 10 minutes. To this solution, sodium
triacetoxyborohydride (635 mg, 3 mmol) was added at once and the reaction
mixture
was stirred at room temperature for another 30 minutes. Progress of the
reaction was
monitored by HPLC and LC/MS. Reaction was completed in 30 minutes. DMF was
then removed and the crude material obtained was then precipitated using
diethyl ether/
MeOH (100/10 mL). Filteration of the precipitate gave a yellow powder, which
was
used for the next step without further purification.
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Step 2:
7-Iodo-9- (3,3,3-trifluoro-propylamino)-methyl-sancycline (665 mg, 1 mmol),
Pd(PPh3)4 (115mg, 0.1 mmol), Pd(OAc)2 (22 mg, "0.1 mmol), CuI (19 mg, 0.1
mmol)
were taken in anhydrous DMF (30 mL) and purged with argon for 5 minutes. To
this
solution, 2-pyrazine-stannane (738 mg, 2 mmol) was added and the reaction
mixture was
stirred at room temperature for 2 hours. Reaction was completed by then
(monitored by
BPLC/LCMS). It was then filtered through celite, washed with 5 mL of methanol.
Solvent was evaporated to dryness. The crude material obtained was purified
using
preparative HPLC. A yellow solid was obtained after evaporating the fractions,
which
was converted to its HCl salt using MeOH/HCI solution. LC-MS (M+1 618).
7-Amino-9-Iodo-Doxycycline
C. CH3
NH2 CF-h N
OH
+/ _ I NH2
OH 0 tH&O 0
To 500 mg of 9-Iodo-doxycycline in 10 ml of methanesulfonic acid was added
1.1 eq. of sodium nitrate. The reaction mixture was left stirring for several
hrs and was
monitored by analytical HPLC. The intermediate (9-Iodo-7-nitro-doxycycline)
was
isolated by diluting he solution with ice-water, adjusting the pH with sodium
hydroxide
(pH -4) and extracting the product with n-butanol. The solvent was evaporated
under
reduced pressure and the crude material was subjected to hydrogenation using
10% Pd/C
in methanol. The final product was obtained via preparative HPLC. The LCMS
showed
the desired material; MS: 586. The structure was confirmed by NMR
7-(Dimethvlamino)-9-(4'.4'-Difluoropiperdin lll-Doxycycline
H3C`N/CH HOH \NCF6
F H OH
F-
N I / _ I NH,
OH 0 OH 0 0
To a solution of 105 mg (0.16 mmol) of 9-(4-difluoropiperdinyl)-doxycycline
dihydrochloride in 10 mL of methanesulfonic acid at room temperature, was
added 19.4
mg (0.19 mmol) of potassium nitrate dissolved in 4 mL of methanesulfonic acid.
The
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reaction was monitored by LCMS. After 30 minutes, the reaction mixture was
poured
over ice and diluted to 160 mL with ice water. The solution was. loaded onto a
2.5 x 1
cm column of divinylbenzene resin (1000 angstrom, 5-25 pm) equilibrated with
water.
The crude reaction mixture was washed with excess water to remove
methanesulfonic
acid followed an excess of IN ammonium acetate to neutralize the crude
mixture. The
excess ammonium acetate was removed by a water wash and the crude compound was
purified by elution with 40% methanol in water with 0.1 % HCL The purified
material
was evaporated to dryness to yield 70 mg of 9-(4-difluoropiperdinyl)-7-nitro-
doxycycline as the dihydrochloride salt (Yield = 63%). LCMS (MH+) 623. To 70
mg
(0.10 mmol) of 9-(4-difluoropiperdinyl)-7 nitro-doxycycline dihydrochloride in
20 mL
of methoxyethanol was added 200 mL of sulfuric acid and 162 mL (2 mmol) of 37%
formaldehyde in water. The reaction mixture was purged with Argon gas and 40
mg of
10% wet Palladium on carbon was added with stirring. The reaction was
hydrogenated
at room temperature and 760 torr hydrogen gas for 12 hours. The crude reaction
was
passed through Celite and evaporated to dryness. The crude reaction mixture
was
purified by preparative HPLC (1 inch x 25 cm, Phenomenex Luna C18, 10 mm,
Gradient 5-40% B buffer, A = water + 0.1% TFA, B = acetonitrile + 0.1 % TEA,
detection at 280 nm) to yield 20 mg of the product as the dihydrochloride salt
(Yield =
30%). LCMS (MH+) 621.
7 Diethvlamino-9-(4'-Fluoro-N-Piperidinyl methyl)-Sancycline
CH3 CH3
LNJ H3C.N.CH3
F H H= OH
N I NHZ
OH O OH O
7 NH2-sancycline (4.0g, 9.32mmol) was combined with 2-methoxyethanol
(100mL), H2SO4 (5mL of IN solution) in a 2 -neck 250mL round bottom flask.
Acetaldehyde (5.2mL, 9.32mmol) was added to reaction solution and contents
were
stirred at room temperature under argon for 20 minutes. Pd/C (1.25g) was added
to
reaction and contents were evacuated/flushed with argon 3 times. A balloon
filled with
H2 was placed on top neck of reaction flask and reaction solution was
evacuated/flushed
with H2 three times. The reaction was stirred overnight under H2 pressure at
room
temperature. The reaction was monitored by HPLC and LCIMS and shown to be
complete by morning. The mixture was filtered through celite and solvent
evaporated in
vacuo. The residue was redissolved in water (IL) and the pH was adjusted with
Et3N to
pH 5. The mixture was filtered again through celite and loaded onto a DVB
column.
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The compound eluted at 15% CH3CN. Clean fractions were evaporated and dried
overnight under vacuum. A yellow/brown solid (7-diethylamino sancycline) was
isolated in 40% yield.
7-diethylamino sancycline (1.4g, 2.88mmol)was dissolved in TFA/Triflie acid
(22mL/6mL) in a 100mL flask. N-iodosuccinimide (12g, 5.78mmol) was added
portionwise to reaction solution every 20 minutes. The reaction monitored by
HPLC
and LCMMS and shown to be complete within 3 hours. The reaction solution was
diluted
with H2O (0.1% TFA) (3OmL) and the solvent was evaporated. The residue was
redissolved in H2O (100mL) and loaded onto a 5g DVB cartridge. The crude
product
eluted at 30-50% CH3CN. A yellow/brown crude product was isolated in 90%
yield.
This crude material, 7-diethylamino-9-iodo-sancycline, (1.8g, 2.95mmol) was
dissolved in anhydrous DMF (IOOmL) in a 2 neck IL round bottom flask and
placed
under argon. NaOAc (0.61g, 7.36mmol) was added to reaction solution and
stirred at
room temperature 45min. Pd(PPh3)4 (1.028, 8.85mmol) was added to reaction and
a
CO-filled balloon was placed on top neck of reaction flask. CO was bubbled
through
reaction solution for 10min. then flask opened to CO balloon. SnBu3H (0.8g,
2.95mmol) was added via syringe pump to reaction solution over 1 hour while
heating to
65 C (oil bath temperature). The reaction was monitored by LCMMS and shown to
be
complete upon addition of tin hydride. H2O (0.1 % TFA, 0.3L) was added to the
reaction
flask and a precipitate formed. The mixture was filtered through celite and
the filtrate
was evaporated in vacuo. A brown solid in 90% yield (crude material) was
isolated.
7-diethylamino-9-formyl-sancycline (0.25g, 0.49mmol) was dissolved in DMF
(lOmL). InCl3 (0.01g, 0.049mmol), 4-fluoropiperidine.HCl (0.15g, 0.98mmol),
and
Et3N (0.09g, 0.98mmol) were added to reaction solution. The reaction was
stirred at
room temperature under argon 45 minutes. NaCNBH3 (0.043g, 0.68mmol) was added
to
the reaction and it was monitored by HPLC and LCMMS. The reaction was shown to
be
complete in 3 hours and it was quenched with McOH (3OmL). The final product
was
isolated by preparative HPLC in 10% yield as a yellow solid. ESI MS: m/z (M +
H)
601.
Synthesis of 7-Aminomethyl Doxycycline
HP HaC~NiOHa
1 H - H -
ON
NH,
OH
OH 0 OH 0 0
To 1 gram of 9-tert-butyl-doxycycline, dissolved in 15 ml of methanesulfonic
acid, was added an excess of HMBC (Hydroxymethyl-carbamic acid benzyl ester).
The
reaction mixture was monitored by analytical HPLC. The LCMS showed MS: 530
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corresponding to the desired material, 7-aminomethyl-9-t-butyl doxycycline.
The
product was isolated via preparative HPLC and the structure confirmed by NMR.
Removal of the t-butyl in triflic acid afforded the 7-aminomethyl doxycycline
in good
yield.
Synthesis of 9-(3'.3'.3'-Trifluoro~ropylamino methyl Minocycline
H3C..N.CH3 H3C.N.CH3
H H OH
F F ~F N NH2
F OH O OH O
9-formyl-minocycline (0.2g, 0.42mmol) was combined with InC13 (0.01g,
0.005mmol), 3,3,3-trifluoropropylamine.HC1(0.25g, 1.7mmol), Et3N (0.17g,
1.7mmol),
and DMF (l OmL) in a glass vial. The reaction was stirred at room temperature
under
argon for 1 hour. NaCNBH3 (0.032g, 0.50mmol) was added to reaction solution
and
was monitored by HPLC and LC/MS. The reaction was complete within 1 hour,
quenched with MeOH (2OmL) and the solvent evacuated in vacuo. The final
product
was isolated by preparative HPLC in 25% yield as a yellow solid. ESI-MS: m/z
(M + H)
583.
9-(4'-Difluoromethylene-N-pineridinyl methyl Minocycline
F H3C,N,CH3 H3C.N,CH3
H H '
F OH
N I / \ _ I NH2
OH O OH
00
o
Anhydrous tetrahydrofuran (THF, 200 mL) was placed in a flame-dried 500 mL
round bottom flask at 0 C in an ice bath. Dibromodifluoromethane (97%,
Aldrich,
10.00 mL, 106.19 mniol, 4.3 eq.) was added via syringe. Ten minutes later,
Hexamethylphosphorous triamide (HMPT, 97%, Aldrich, 19.50 n2I,104.07 mmol, 4.2
eq.) was added dropwise. The clear solution turned milky white and was stirred
for I
hour at 0 C. A solution of tent Butyl 4-oxo-l-piperidinecarboxylate (98%,
Aldrich,
5.00 g, 24.59 mmol, 1.0 eq.) in anhydrous THE (50 mL) was then added dropwise
via
syringe at 0 C and the solution was allowed to warm up slowly to room
temperature
over 1 hour by removing the ice bath The powdered zinc (99.998%, Aldrich,
powdered,
-100 mesh, 6.56 g, 98.34 mmol, 4.0 eq.) was then added followed by HMPT (1.15
mL,
6.14 mmol, 25%) and the reaction mixture was refluxed for 3 hours. Water (250
mL)
and Diethyl ether (Et20, 250 mL) were added and the mixture was extracted with
Et2O
(3 times 100 mL). The combined organic layers were washed with a saturated
solution
of Copper(fl) sulfate (CuSO4) in Water (150 mL) then with water (150 mL). The
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organic layer was dried over Magnesium sulfate (MgSO4), filtered, and
evaporated
under reduced pressure to yield the desired fluorinated piperidine as a yellow
oil, which
was used without further purification in the next step.
A 100 mL round bottom flask equipped with a magnetic stirring bar was loaded
with the BOC-protected piperidine (2.00 g, 8.57 mmol, 1.0 eq.) in a saturated
HC1
solution in Methanol (50 mL) at room temperature. The mixture was then stirred
at
40 C for 30 minutes and the solvent was evaporated under reduced pressure to a
minimal volume. The HCl salt was then precipitated from Et2O, filtered, and
dried in
vacuo to yield the desired fluorinated piperidine (1.10 g, 6.49 mmol, 76%
yield) as a
beige solid used without further purification in the next step.
A flame-dried 50 mL round bottom flask equipped with a magnetic stirring bar
was loaded with 9-Formyl-minocycline (500 mg, 1.03 mmol,1.0 eq.) in anhydrous
Dimethylformamide (DMF, 10.00 mL) at room temperature. Indium chloride (InC13,
99.999%, Aldrich, 59 mg, 0.27 mmol, 26%) was added and the reaction mixture
was
stirred at 30 C for 10 minutes. The amine (350 mg, 2.06 mmol, 2.0 eq.) was
added in
anhydrous DMF (2 mL), followed by Triethylamine (NEt3, 99.5%, Alfa-Aesar, 290
L,
2.08 mmol, 2.0 eq.). The mixture was then stirred at 30 C for 1 hour and
Sodium
triacetoxyborohydride (NaBH(OAc)3, 95%, Aldrich, 220 mg, 1.04 mmol, 1.0 eq.)
was
added followed by more NEt3 (300 L). After 2 hours, the reaction was done and
the
solvent evaporated under reduced pressure. The residue was purified by
preparative
BPLC (Acetonitrile / Water / 0.1% Trifluoroacetic acid gradient) to yield the
desired
product as a yellow solid. MS m/z 603.
Synthesis of 9-(4'-Fluoro-N-PiperdinyI) methyl Doxycycline
?aH O~H~uia'~a
F~ " - OH
H .
OH
OH O OH 0 0
The compound was prepared from Doxycycline (2.5g, 5.0mmol) dissolved in
MeOH (anhydrous) (25mL) and combined with AgSO4 (3.7g, 1lmmol) and 12 (3.1g,
11mmol) in a 100mL round bottom flask. H2SO4rr, (2 drops) was added to the
reaction
solution and stirred at room temperature under argon for 1 hour. The reaction
solution
turned bright yellow after 30 minutes and the reaction was monitored by LC/MS
and
shown to be complete in 1 hour. Sodium sulfite (sat) (8mL) was added to the
reaction
solution and a thick yellow precipitate was formed. The mixture was stirred at
room
temperature for 20 minutes. The mixture was diluted with CH3CN (75mL),
filtered
through celite and evaporated solvent in vacuo to yield 1.7g of crude 9-iodo-
doxycycline
material.
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9-iodo-doxycycline (1.3g, 2.4mmol) was dissolved in anhydrous DMF (2OmL) in
a 200mL 2 neck round bottom flask and Pd(PPh3)4 (0.82g, 0.71mmol) was added. A
CO-filled balloon was placed on top neck of reaction flask and CO was bubbled
directly
into reaction from lecture bottole. The flask was then opened to the balloon
and SnBu3H
(0.70g, 2.7mmol) was added via syringe pump over 1 hour. The reaction solution
was
heated to 65 C during the tin addition. The reaction was monitored by LCIMS
and it
was shown to be complete once the tin addition was complete. Water (0.1 % TFA)
(200mL) was then added to reaction solution and a yellow precipitate formed.
The
mixture was then filtered through celite and the filtrate was evaporated in
vacuo. A
brown/yellow solid in 50% yield was isolated.
(9-formyl-doxycycline (0.20g, 0.42mmol) combined with InC13 (0.01g,
0.042mmol), 4-fluoropiperidine (0.13g, 0.84mmol), Et3N (0.09g, 0.84mmol), and
DMF
(5mL) in a glass vial. The mixture was stirred under argon at room temperature
for 30
minutes. NaCNBH3 (0.037g, 0.59mmol) was added to the reaction vial and the
reaction
continued to be stirred at room temperature under argon. The reaction was
monitored by
LC/MS and HPLC and shown to be complete after 1 hour. The reaction was
quenched
with MeOH (15mL) and the solvent was evacuated in vacuo. The product was
isolated
by preparative HPLC in 10% yield as a yellow solid. ESI-MS: m/z (M + H) 559.
Synthesis of 9-(Benzyl methyl-amino)-Propynyl)-Minocycline
H3C.N,CH3 HSC..N..CH3
H H OH
QJH,4~)~;NOH O OH O
7-Iodo-minocycline (1.08 g, 1.86 mmol ), taken in 25 mL of acetonitrile was
degassed and purged with nitrogen (three times). To this suspension Pd(OAc)2
(20 mg,
.089 mmol), CuI (10 mg, .053 mmol), (o-tolyl)3P (56 mg, .186 mmol) were added
and
purged with nitrogen for few minutes. Benzyl-methyl-prop-2-ynyl-amine (318 L,
2
mmol) and triethylamine (1 mL) were added to the suspension. It turned into a
brown
solution after the addition of Et3N. The reaction mixture was then heated to
70 C for 2
hours. The progress of the reaction was monitored by HPLC/LCMS. It was then
cooled
down to room temperature and was filtered through celite. Evaporation of the
solvent
gave a brown solid, which was then purified on preparative HPLC to afford the
desired
compound. LC-MS (M+1615).
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Synthesis of 8-(2'-1(2'-Fluoro-eth)lamino)-methyll-phen +y )1-Sancycline
H3C. CH3
H H N' OH
F~~N I c _ I NH2
H OH 0 OHO O O
Step 1:
To a stirred solution (cooled at 00 C, ice-bath) of 9-amino-sancycline (7 g,
16.3
mmol) in 200 mL of McOH, 48% HBF4 solution (5.32 mL, 40.75 mmol) was added
slowly under an argon atmosphere. After 5 minutes, n-BuNO2 (2.1 MI, 17.93
mnaol )
was added slowly (dropwise). The reaction mixture was then stitrred at 0 C for
3 hours
(monitored by HPLC/LC-MS). NaN3 (1.06g, 16.3 mmol) was then added the reaction
mixture (all at once). The reaction mixture was stirred at 0 C for another 3
hours
(monitored by BPLC/LC-MS). The reaction mixture was then poured slowly into
stirring diethyl ether (1L at ice-bath temperature). A yellow precipitate was
obtained
and it was filtered, washed with ether (20m1 x 3) and dried under vaccum,
sealed in a
vial and stored at 0 C. Isolated yield 7g.
Step 2:
Hydrobromic acid (30% in acetic acid) (14 mL) was added to a flask and cooled
to 0 C. 9-Azido-sancycline (1g, 2.2 mmol) was added to the flask and the
reaction was
left to stir for one hour. After 1 hour, the reaction was complete. The
reaction mixture
was precipitated in 300 mL of diethyl ether. After letting the solution
settle, the top
layer of diethyl ether was decanted and the reaction mixture was dried under
vaccum. A
brown-black solid was then dissolved in methanol and precipitated using
diethyl ether.
The solid obtained was filtered and dried under vaccum.
Step 3:
To a stirred solution (cooled at 0 C, ice-bath) of 8-bromo-9-amino-sancycline
(828 mg, 1.6 mmol) in 200 mL of McOH, 48% HBF4 solution (0.53 mL, 4.0 mmol)
was
added slowly under an argon atmosphere. After 5 minutes, n BuNO2 (0.2 mL, 1.79
nnnol ) was added slowly (dropwise). The reaction mixture was then stitrred at
0 C for
2 hours and left overnight at room temperature (monitored by HPLC/LC-MS). The
solvent was evaporated and the crude material obtained was precipitated using
diethyl
ether (300 mL). The solid obtained was filtered and dried under vaccum.
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Step 4:
8-Bromo-sancycline (492 mg, 1 mmol) and Pd(OAc)2 (22 mg, 0.1 mmol) were
taken in methanol (150 mL) and purged with argon while heating the reaction
mixture at
65 C (oil bath temperature). After 10 minutes, an aqueous solution of sodium
carbonate
(315 mg, 3 mmol in 10 mL of water) was added. A yellow precipitate was
obtained which
was fisher heated for another 10 minutes, before adding a DMF solution of the
boronic
acid( 300 mg, 2 mmol in 10 mL of DMF). The reaction was then heated at 65 C
for 3
hours. The reaction was monitored by BPLC/LCMS. The mixture was cooled down to
room temperature and then filtered through celite. The solvent was then
evaporated and
the crude materialobtained was precipitated using methanol/diethyl ether
(10/200 mL).
The crude material was then filtered and dried under vacuum. The yellow-brown
material
obtained was used as such without further purification.
Step 5:
To a solution of 8-(2-formyl-phenyl)-sancycline (518 mg, 1 mmol) in 30 mL of
DCE under an argon atmosphere, 2-fluoro-ethylamine hydrochloride (198 mg, 2
mmol)
and triethylamine (202 L, 2 mmol) were added. The reaction mixture was then
stirred
at room temperature for 2 hours. The reaction was monitored by using
HPLC/LCMS,
and was completed in 2 hours. The solvent was then evaporated and the crude
material
was purified using preparative HPLC to afford the desired compound. LC-MS (M+1
566).
7-Pyrazolyl-S ancycline
H
N-N
ENH2
OH O OHO 0
To a stirred solution of 7-Iodo sancycline (100 mg, 0.153 mmol) in DMF (1 mL)
was added pyrozole-4-boronic acid pinacole cyclic ester (77 mg, 0.40 mmol),
methanol
(1.5 mL), tetrakis(triphenylphosphine)palladium (18 mg, 0.015 mmol) and a
solution
containing 250 mg CsCO3 in 0.7 mL water. The reaction mixture was then subject
to
microwave irradiation at a temperature of 100 C for 5 minutes. The reaction
was then
diluted with 100 mL of water and TFA was used to lower the pH to 2. This
solution was
then filtered through celite, and loaded onto a plug of divinyl benzene resin
(DVB). The
plug containing the product was washed with water (200 mL) before the final
compound
was eluted with MeCN and reduced by rotary evaporation. The crude material was
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purified by reverse phase HPLC to give the final product (64 mg, 0.12 mmol,
75% yield)
LCMS m/z=481.2115 (M+H).
Synthesis of 9-[(2,2,2-Trifluoro-ethyl)-hydrazonomethylj-Minocycline
H3C1 N,CH3 H,C,N,CH3
H H OH
F~'H NI _ NHZ
F OH O OH O
To a solution of 9-formyl minocycline (485 mg, 1 mmol) in 30 mL of DMF
under an argon atmosphere, indium trichloride (22 mg, 0.1 mmol) and
trifluoroethylhydrazine (228 L, 2 mmol) were added. The reaction mixture was
then
stirred at room temperature for 30 minutes. The reaction was monitored by
using
HPLC/LCMS, and was completed in 30 minutes. The solvent was then evaporated
and
the crude material was purified using preparative HPLC to afford the desired
compound.
LC-MS (M+1 582).
Synthesis of 9-(l'-Isopropy1-4'-piperidinyl) amino Sancycline
CH3 . H3C.N.CH3
~N H Fi OH
H 3 C \
N NHZ
OH O OHO O O
To a solution of 9-amino sancycline HCl salt (0.5g, lmmol) in 40 ml of
methanol
and was added 1-isopropyl-4-piperidone (0.14g, 2 mmol). The solution was
stirred for
5 minutes at room temperature. Sodium cyanoborohydride (62.5 mg, 1 mmol) was
introduced, followed by the addition of 4 ml of AcOH. The mixture was stirred
at room
temperature for 1 hour until all starting material disappeared. The suspension
was
filtered and purified by HPLC to afford the title compound (210mg). LC-MS (M+1
555).
S3mjkesis of 9-(3-t-butyl-N inudazolvl)-methy1)-Minocycline
H3C. N CH3 H3C.N CH3
H OH
H3C{N-
N NHZ
H3C
OH O W 0
To a stirred solution of 9-aminomethyl- minocycline (2.50 g, 4.14 mmol) in
DMF (25 mL) and MeOH (15 mL) was added 1-bromopinacolone (1.34 mL, 1.01 g,
5.63 mmol) and Cs2CO3 (5.0 mL of a IN aqueous solution, 5.0 mmol). The
reaction
was heated to 100 C for 15 minutes in a pressure vesicle using microwave
irradiation.
The contents were then diluted with water (1.0 L) and Na2CO3 was used to
adjust the pH
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to 6. This solution was then filtered through celite and loaded onto a plug of
divinyl
benzene resin. The product was washed with water (500 mL) before it was eluted
with
MeCN and reduced by rotary evaporation. The crude material was purified by
reverse
phase HPLC to give the tert-butyl-ketone intermediate (680 mg, 1.90 mmol, 50%
yield).
To a stirred solution of the tert-butyl-ketone intermediate (68 mg, 0.190
mmol) in
formamide (1.0 mL) was added triethyl-amine (0.020 mL, 28 mg, 0.27 mmol) to
adjust
the pH to 8. The reaction was heated to 100 C for 5 minutes in a pressure
vesicle using
microwave irradiation. The contents were then diluted with water (100 mL) and
TFA
was used to adjust the pH to 2. This solution was then filtered through
celite, and loaded
onto a plug of divinyl benzene resin. The product was washed with water (200
mL)
before it was eluted with MeCN and reduced by rotary evaporation. The crude
material
was purified by reverse phase HPLC to give the final compound (6.0 mg, 10
mol, 4%
yield) LCMS m/z = 594.4863 (M +H).
Synthesis of 9-(2-thiol 5-methyl-N-imidazolyll-methyl Minocycline
H3C.N.CH3 HaC,N.CH3
H H OH
~49XNH2
H3C OH 0 01-P 0
To a stirred solution of 9-am inomethyl-minocycline (2.00 g, 4.12 mmol) in DMF
(12 mL), MeOH (6.0 nL) and acetic acid (3.0 nL) was added KSCN (0.400 g, 4.12
mmol) and Acetol 0.400 mL, 0.370 g, 5.00 mmol). The reaction was heated to 100
C
for 15 minutes in a pressure vesicle using microwave irradiation. The contents
were
then diluted with water (1.0 L) and Na2CO3 was used to adjust the pH to 6.
This
solution was then filtered through celite and loaded onto a plug of divinyl
benzene resin.
The product was washed with water (500 mL) before it was eluted with MeCN and
reduced by rotary evaporation. The crude material was purified by reverse
phase HPLC
to give the final product (620 mg, 1.06 mmol, 26% yield) LCMS m/a = 584.3998
(M +
H).
Synthesis of 7-(2',2'-dimethyl-propyl)amino methyl Sanevcline
H3C CH3HH
H3C.N.CH3
H3C H H - OH
NH2
OH O OHO O O
1 g of 7-aminomethyl-sancycline, 3 equivalents of trimethylacetaldehyde and
one equivalent of indium trichloride were dissolved in 10 ml of DMF. The
mixture was
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stirred at room temperature for 15 minutes. To this mixture was added 3
equivalents of
sodium triacetoxyborohydride. The resulting reaction mixture was left stirring
for
several hours. The reaction was monitored by analytical HPLC. The LCMS showed
MS: 514 which corresponds to the desired material. The product was isolated
via
preparative HPLC and the structure was confirmed by NMR
Synthesis of 9-Benzoimidazoly1l-Minoce
H3C.N.CH3 H3C.N.CH3
H H OH
N~ I / _ I NH,
NH OH O OH O
To a stirred solution of the trifluoroacetic acid (TFA) salt of 9-formyl
minocycline (488 mg, 1.47 mmol) in DMF (3 mL) and MeOH (2 mL) was added 1,2-
phenylenediamine (80 mg, 0.74 mmol). The reaction was heated to 50 C and was
complete in 5 minutes. The contents were then diluted with water (500 mL) and
TFA
was used to adjust the pH to 2. This solution was then filtered through
celite, and loaded
onto a plug of divinyl benzene resin. The plug containing the product was
washed with
water (300 mL) before it was eluted with MeCN and reduced by rotary
evaporation.
The crude material was purified by reverse phase HPLC to give the
Benzoimidazol
product (100 mg, 0.175 mmol, 10% yield) LCMS m/z = 574.3637 (M + H).
Example 2: In vitro Minimum Inhibitory Concentration (NIIC) Assay
The following assay is used to determine the efficacy of the tetracycline
compounds against common bacteria. 2 mg of each compound is dissolved in 100
l of
DMSO. The solution is then added to cation-adjusted Mueller Hinton broth
(CAMHB),
which results in a final compound concentration of 200 g per ml. The
tetracycline
compound solutions are diluted to 50 pL volumes, with a test compound
concentration
of .098 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
1x106 CFU/ml. At OD=1, cell densities for different genera should be
approximately:
E. coli - 1x109 CFU/ml
S. aureus 5x10$ CFU/ml
Enterococcus sp. 2.5x 109 CFU/ml
50 l of the cell suspensions are added to each well of microtiter plates.
The final cell density should be approximately 5x105 CFU/ml. These plates are
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WO 2005/009943 PCT/US2004/020249
incubated at 35 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 tetracycline compound that inhibits
growth.
EQUIVALENTS
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 appropriate components, processes, and methods of those patents,
applications and
other documents may be selected for the present invention and embodiments
thereof.
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